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Articles by Charity A. Dunn in JoVE

 JoVE Biology

A Fluorescent Screening Assay for Identifying Modulators of GIRK Channels

1Department of Pharmacology, Physiology & Neuroscience, University of South Carolina, School of Medicine


JoVE 3850

A real-time screening procedure for identifying drugs that interact with G protein-gated inward rectifier K+ (GIRK) channels is described. The assay utilizes membrane potential-sensitive fluorescent dyes to measure GIRK channel activity. This technique is adaptable for use on a number of cell lines.

Other articles by Charity A. Dunn on PubMed

Depth Dependence of Vascular Fluorescence Imaging

In vivo surface imaging of fluorescently labeled vasculature has become a widely used tool for functional brain imaging studies. Techniques such as phosphorescence quenching for oxygen tension measurements and indocyanine green fluorescence for vessel perfusion monitoring rely on surface measurements of vascular fluorescence. However, the depth dependence of the measured fluorescence signals has not been modeled in great detail. In this paper, we investigate the depth dependence of the measured signals using a three-dimensional Monte Carlo model combined with high resolution vascular anatomy. We found that a bulk-vascularization assumption to modeling the depth dependence of the signal does not provide an accurate picture of penetration depth of the collected fluorescence signal in most cases. Instead the physical distribution of microvasculature, the degree of absorption difference between extravascular and intravascular space, and the overall difference in absorption at the excitation and emission wavelengths must be taken into account to determine the depth penetration of the fluorescence signal. Additionally, we found that using targeted illumination can provide for superior surface vessel sensitivity over wide-field illumination, with small area detection offering an even greater amount of sensitivity to surface vasculature. Depth sensitivity can be enhanced by either increasing the detector area or increasing the illumination area. Finally, we see that excitation wavelength and vessel size can affect intra-vessel sampling distribution, as well as the amount of signal that originates from inside the vessel under targeted illumination conditions.

A Comparison of Extraperitoneal and Intraperitoneal Approaches for Robotic Prostatectomy

Objectives. This study compared oncologic and health-related quality-of-life outcomes among patients undergoing intraperitoneal or extraperitoneal robotic prostatectomy. Methods. Of 215 patients undergoing robotic prostatectomy, the approach was intraperitoneal in 48 and extraperitoneal in 167. Cancer control was evaluated using margin status. Recovery after surgery and functional health was assessed using the Convalescence and Recovery Evaluation and Expanded Prostate Cancer Index Composite questionnaires, respectively. Results. Positive surgical margin rates were similar between approaches (14% extraperitoneal, 10% intraperitoneal; P = .63). Functional outcomes were slightly improved for those with the extraperitoneal approach (ie, higher urinary irritation/obstruction scores at 3 months). The extraperitoneal group demonstrated higher activity (91.8 vs 83.3, P = .03) and cognitive scores (94.9 vs 91.7, P = .04) at 6 weeks as well as higher gastrointestinal scores at 2 weeks (94.2 vs 90.8, P = .05). Conclusions. These data support efforts to broaden the adoption of the extraperitoneal approach for robotic prostatectomy.

Continuous Intercostal Nerve Blockade for Rib Fractures: Ready for Primetime?

Providing analgesia for patients with rib fractures continues to be a management challenge. The objective of this study was to examine our experience with the use of a continuous intercostal nerve block (CINB). Although this technique is being used, little data have been published documenting its use and efficacy. We hypothesized that a CINB would provide excellent analgesia, improve pulmonary function, and decrease length of stay (LOS).

Rural Kenyan Men's Awareness of Danger Signs of Obstetric Complications

For many women in Kenya, their husbands act as gate-keepers to access of healthcare services. Awareness of the danger signs of obstetric complications is the essential first step in accepting appropriate and timely referral to obstetric care. The objectives of this study were to assess men's awareness of the danger signs of obstetric complications, and to identify any associated demographic factors.

Should Sex-ratio Distorting Parasites Abandon Horizontal Transmission?

Sex-ratio distorting parasites are of interest due to their effects upon host population dynamics and their potential to influence the evolution of host sex determination systems. In theory, the ability to distort host sex-ratios allows a parasite with efficient vertical (hereditary) transmission to dispense completely with horizontal (infectious) transmission. However, recent empirical studies indicate that some sex-ratio distorting parasites have retained the capability for horizontal transmission.

Developmental Delays in Children With Neurofibromatosis Type 1

It is well documented that children with neurofibromatosis type 1 are at high risk for a variety of cognitive and learning deficits. The current study investigated the use of a developmental screening tool, the Parents' Evaluation of Developmental Status: Developmental Milestones, as an accurate, reliable, and efficient indicator of developmental delays. Sixty-eight percent of children with neurofibromatosis type 1 were found to have a developmental delay in at least 1 of the 8 areas tested by the Parents' Evaluation of Developmental Status: Developmental Milestones. Significant developmental abnormalities were found in the areas of fine motor (35%), gross motor (52%), and math/premath (31%). A positive association was found between the presence of a previously diagnosed optic glioma and math/premath delays (χ(2) = 0.0022) and between male sex and fine motor delays (χ(2) = 0.0325). The Parents' Evaluation of Developmental Status: Developmental Milestones assessment demonstrates the high presence of developmental delays in children with neurofibromatosis type 1 and the need for aggressive and early screening.

A Five-year Prospective Study of Quality of Life After Colorectal Cancer

PURPOSE: Long-term (≥5 years) quality of life after colorectal cancer is not well described. The present study assessed quality of life (QOL) and psychological distress in colorectal cancer survivors more than 5 years to describe changes over time and antecedents of long-term outcomes. METHOD: A prospective survey of a population-based sample of 763 colorectal cancer patients assessed socio-demographic variables, health behaviors, optimism, threat appraisal, and perceived social support at 5 months post-diagnosis as predictors of QOL and psychological distress 5 years post-diagnosis. RESULTS: QOL improved over time (P < 0.01 for each measure); however, measures of psychological distress remained stable (P > 0.07 for each measure). Risk factors for poorer QOL and/or greater psychological distress included: later stage disease, having a permanent stoma, rectal cancer, fatigue, smoking, being single, low social support, low optimism, and a more negative cancer threat appraisal. Being women, having a pet, having a private health insurance, and receiving both surgery and adjuvant treatment were protective. CONCLUSION: Consistent with response shift theory, the antecedents of QOL after colorectal cancer are multifactorial and include predisposing socio-demographic, medical, and psychological variables. Psychosocial interventions that target both social support and threat appraisal may be effective for this patient group. Additional stepped-up support may be needed for people from a poorer social environment who have multiple risk factors for poorer adjustment. Health system effects require further investigation.

Emerging Psychoactive Substance Use Among Regular Ecstasy Users in Australia

BACKGROUND: The past decade has seen the development of an array of emerging psychoactive substances (EPS), however, there is minimal information on the extent of their use outside Europe. This study aimed to determine the extent of use of EPS from stimulant (such as mephedrone) and psychedelic classes (such as 5-methoxy-dimethyltryptamine [5-MeO-DMT]) among an Australian sample of regular ecstasy users (REU). Further, to determine if consumers of these drugs represent a distinct subgroup of REU. METHODS: Australian national cross-sectional surveys of 693 regular (at least monthly) ecstasy users conducted during 2010. RESULTS: More than one quarter (28%) of REU had used an EPS in the past six months, most commonly from the stimulant class (20%, typically mephedrone, 17%) rather than the psychedelic class (13%). Demographics and risk behaviours of REU that used stimulant EPS were largely no different from non-EPS consuming REU. Those using psychedelic EPS were distinct, initiating ecstasy use earlier, more frequently using multiple substances (cannabis, inhalants, GHB, ketamine) and more commonly experiencing legal, psychological and social problems. CONCLUSIONS: Psychedelic EPS use appears largely restricted to a distinct subset of REU with high-level non-injecting polydrug use, but use appears generally limited. The demographic similarity of stimulant EPS consumers with 'mainstream' REU, in conjunction with positive responses to the psychoactive effects of these drugs and declining ecstasy purity, suggests strong potential for stimulant EPS to expand further into ecstasy markets. Such drugs may have a greater public health impact than ecstasy, and merit careful monitoring into the future.

Development and Usability Testing of a Parent Decision Support Tool for the Neonatal Intensive Care Unit

In this paper we present the development and evaluation of a parent decision support tool for a neonatal intensive care unit (NICU), known as PPADS or Physician and Parent Decision Support. The NICU interprofessional (IP) team uses advanced technology to care for the sickest infants in the hospital, some at the edge of viability. Many difficult care decisions are made daily for this vulnerable population. The PPADS tool, a computerized decision support system, aims to augment current NICU decision-making by helping parents make more informed decisions, improving physician-parent communication, increasing parent decision-making satisfaction, decreasing conflict, and increasing decision efficiency when faced with ethically challenging situations. The development and evaluation of the PPADS tool followed a five step methodology: assessing the clinical environment, establishing the design criteria, developing the system design, implementing the system, and performing usability testing. Usability testing of the PPADS tool with parents of neonates who have graduated (survived) from a tertiary level NICU demonstrates the usefulness and ease of use of the tool.

Measurements and Modeling of Transient Blood Flow Perturbations Induced by Brief Somatosensory Stimulation

Proper interpretation of BOLD fMRI and other common functional imaging methods requires an understanding of neurovascular coupling. We used laser speckle-contrast optical imaging to measure blood-flow responses in rat somatosensory cortex elicited by brief (2 s) forepaw stimulation. Results show a large increase in local blood flow speed followed by an undershoot and possible late-time oscillations. The blood flow measurements were modeled using the impulse response of a simple linear network, a four-element windkessel. This model yielded excellent fits to the detailed time courses of activated regions. The four-element windkessel model thus provides a simple explanation and interpretation of the transient blood-flow response, both its initial peak and its late-time behavior.

Reverse Total Shoulder Arthroplasty in Patients with Parkinson Disease: a Case Series and Review of the Literature

Parkinson disease (PD) is a chronic degenerative neurologic disorder with both motor and nonmotor facets. The motor symptoms, including increased risk for falls, fractures, and stiffness, contribute to the morbidity of arthroplasty. In this article, we report 3 cases of reverse total shoulder arthroplasty in patients with PD. All patients achieved poor functional outcomes with mean (range) active forward flexion of 40° (20°-60°) at follow-up. Although each patient obtained significant pain relief-mean (range) visual analog scale score was less than 1 (0-2)-range of motion was poor. In addition, each patient developed significant glenoid notching, though no component loosening or migration was observed. Mean (range) postoperative follow-up was 17 (4-32) months. A patient who has PD and requires an inverse arthroplasty should be counseled that pain relief may be reliably achieved, while functional outcomes are poor.

Cytomorphology of Erdheim-Chester Disease Presenting As a Retroperitoneal Soft Tissue Lesion

Erdheim-Chester disease (ECD) is a rare, multisystem disorder of macrophages. Patients manifest with histiocytic infiltrates that lead to xanthogranulomatous lesions in multiple organ systems. The cytologic features of this disorder are not well characterized. As a result, the cytologic diagnosis of ECD can be very challenging. The aim of this report is to describe the cytomorphology of ECD in a patient presenting with a retroperitoneal soft tissue lesion. A 54-year-old woman with proptosis and diabetes insipidus was found on imaging studies to have multiple intracranial lesions, sclerosis of both femurs and a retroperitoneal soft tissue mass. Fine needle aspiration (FNA) and a concomitant core biopsy of this abnormal retroperitoneal soft tissue revealed foamy, epithelioid and multinucleated histiocytes associated with fibrosis. The histiocytes were immunoreactive for CD68, CD163, Factor XIIIa and fascin, and negative for S100, confirming the diagnosis of ECD. ECD requires a morphologic diagnosis that fits with the appropriate clinical context. This case describes the cytomorphologic features of ECD and highlights the role of cytology in helping reach a diagnosis of this rare disorder.

The Prevalence of Darunavir-associated Mutations in HIV-1-infected Children in the UK

BACKGROUND: We examined the prevalence of ritonavir-boosted darunavir (DRV) resistance-associated mutations (RAMs) in HIV-infected children in the UK to determine the drug's potential clinical utility as a first-line or second-line protease inhibitor (PI). METHODS: The prevalence of DRV RAMs, identified from IAS 2010 and Stanford, and the Stanford susceptibility score, were estimated in PI-naive and PI-experienced children in the Collaborative HIV Paediatric Study and the UK HIV Drug Resistance Database 1998-2008. Associations between type/duration of PI exposure and area under the viraemia curve on PI with the number of RAMs were investigated using multivariate Poisson regression. RESULTS: A total of 17/417 (4%) children with a resistance test when PI-naive had one IAS DRV RAM, and 1 had a Stanford mutation; none had multiple DRV RAMs. A total of 177 PI-experienced children had a test after a median 2.7 years (IQR 1.1-5.2) on PIs; 19 (11%) had one IAS DRV RAM, 7 (4%) had two RAMs, 1 (0.6%) had three RAMs and 1 (0.6%) had four RAMs. DRV RAMs were independently associated with increased years on a PI, a larger area under the viraemia curve since starting PIs, and any exposure to PIs other than lopinavir (all P≤0.05). Only 6 (3%) PI-experienced children had intermediate-level DRV/ritonavir resistance; none had high-level resistance. CONCLUSIONS: DRV resistance was negligible in PI-naive children and those with lopinavir PI exposure alone. However resistance increased with increasing time, and with higher levels of viraemia, on PIs. Once-daily DRV/ritonavir would be valuable as a second PI or an alternative first PI, particularly if coformulated with a booster in an appropriate formulation for children.

Revision ACL Reconstruction Outcomes: MOON Cohort

Many clinicians believe that the results of revision anterior cruciate ligament (ACL) reconstruction compare unfavorably with primary ACL reconstruction. However, few prospective studies have evaluated revision ACL reconstruction using validated patient-based metrics. This study was performed to evaluate and compare the results of revision ACL reconstruction and primary ACL reconstruction. The Multicenter Orthopaedic Outcomes Network consortium is an NIH-funded, hypothesis-driven, multicenter prospective cohort study of patients undergoing ACL reconstruction. All patients preoperatively complete a series of validated patient-oriented questionnaires. At scheduled 2-year follow-up all patients are given the same series of questionnaires to complete. The study evaluated the results of 2-year follow-up of revision ACL reconstruction performed in 2001. Parameters evaluated included Marx activity level, Knee Injury and Osteoarthritis Outcome Score (KOOS), and International Knee Documentation Committee (IKDC) scores. For this study 446 subjects met inclusion criteria; 2-year follow-up was obtained on 393 (88%). The study group consisted of 55% males with median age of 22 years. There were 33 revision ACL reconstruction cases, for which follow-up was available for 29 (88%). Median baseline Marx (interquartile range) was 12 (8 to 16) and 12 (6 to 16) for the primary ACL reconstruction and revision ACL reconstruction groups, respectively (p= 0.81). At 2 years, median Marx was 9 (4 to 13) and 5 (0 to 10) for the primary ACL reconstruction and revision ACL reconstruction groups, respectively (p= 0.03). Median 2-year IKDC was 75.9 (revision) versus 83.9 (primary) (p=0.003). Median KOOS subscale Knee Related Quality of Life (KRQOL) at 2 years was 62.5 (revision) versus 75 (primary) (p < 0.001), subscale Sports and Recreation was 75 (revision) and 85 (primary) (p = 0.005), subscale Pain was 83.3 (revision) and 91.7 (primary) (p= 0.002). Marx activity score declined at 2-year follow-up in revision ACL reconstruction compared with primary ACL reconstruction. IKDC and KRQOL were significantly decreased in revision ACL reconstruction compared with primary ACL reconstruction at 2-year followup. Revision ACL reconstruction resulted in a significantly worse outcome as measured by these patient-based measures at 2 years.

Comparison of Flexural Properties and Surface Roughness of Nanohybrid and Microhybrid Dental Composites

Recently introduced nanohybrid dental composites have promised a smoother surface finish and strength, comparable to that of microhybrid composites. This study compared the mechanical properties and surface finish of nanohybrid and microhybrid composites by measuring the flexural strength and modulus (four-point bend) and surface roughness after polishing (using atomic force microscopy) of six commercial dental composites (three nanohybrid, three microhybrid). Scanning electron microscopy (SEM) was used to qualitatively characterize filler morphology and size. The flexural strength and modulus were significantly higher among the microhybrid composites, while the nanohybrid composites exhibited significantly lower surface roughness. SEM characterization revealed differences in filler particle size and shape that could affect the flexural properties and surface roughness. Composites containing spherical filler particles exhibited higher flexural properties and lower roughness values compared to composites with irregular fillers. These results did not support the premise that nanohybrid composites offer similar mechanical properties to microhybrids in addition to a better surface finish.

Situations Matter: Teaching the Lewinian Link Between Social Psychology and Rehabilitation Psychology

A little-recognized fact is that social psychology and rehabilitation psychology share a common theoretical ancestry in the situation perspective of Kurt Lewin. Theory and research in both fields assumes that situational influences often override the impact of personal factors, including dispositions. Situational analyses led to the development of a variety of cognitive explanations capturing people's phenomenal accounts for the causes of behavior and concomitant interpretation of social problems. Teachers can explore reasons why, despite the fields' having a shared theoretical perspective and topics of common interest (e.g., attitudes, prejudice, discrimination), little scholarly intradisciplinary contact currently occurs between them.

Community Perspectives on Post-Katrina Mental Health Recovery in New Orleans

Disaster-affected communities may face prolonged challenges to community-wide mental health recovery due to limitations in local resources, infrastructure, and leadership. REACH NOLA, an umbrella non-profit organization comprising academic institutions and community-based agencies, sought to promote community recovery, increase mental health service delivery capacity, and develop local leadership in post-Katrina New Orleans through its Mental Health infrastructure and Training Project (MHIT). The project offered local health service providers training and follow-up support for implementing evidence-based and new approaches to mental health service delivery. This commentary shares the perspectives of three community leaders who co-directed MHIT. They describe the genesis of MHIT, the experience of each agency in adopting leadership roles in addressing post-disaster needs, challenges and growth opportunities, and then overarching lessons learned concerning leadership in a prolonged crisis. These lessons may be relevant to community agencies addressing hurricane recovery in other areas of the Gulf States as well as to inform long-term disaster recovery efforts elsewhere.

Designing Trials of Complex Interventions for Efficacy and Mechanisms Evaluation

Gallbladder Sludge on Ultrasound is Predictive of Increased Liver Enzymes and Total Bilirubin in Cats

The purposes of this retrospective study were to assess the prevalence of gallbladder sludge (GBS) in a population of cats presented for abdominal ultrasound in a teaching hospital and to determine its association with increased serum alanine aminotransferase (ALT), alkaline phosphatase (ALP), and total bilirubin (TB). Gallbladder sludge was detected in 152 (14%) of the cats undergoing abdominal ultrasound between 2004 and 2008. This population was compared to a control group of 32 cats without GBS. Alanine aminotransferase, ALP, and TB mean values were significantly higher in cats with GBS than in controls (P ≤ 0.0005) and odds for increased values in cats with GBS were 4.2 [95% confidence interval (CI): 1.6 to 11.0], 9.5 (95% CI: 2.2 to 41.7), and 4.1 (95% CI: 1.5 to 11.5), respectively (P ≤ 0.007). In conclusion, GBS is an uncommon ultrasonographic finding in cats that is predictive of increased liver enzymes and TB. More studies are needed to establish potential links between GBS and hepatobiliary disease in cats.

A Pilot Study of Rhythm and Timing Training As a Supplement to Occupational Therapy in Stroke Rehabilitation

Stroke is the leading cause of disability. A need exists for an effective intervention to enhance upper extremity (UE) motor abilities and activities of daily living (ADL) performance.

Eye Health Service Access and Utilization in the National Indigenous Eye Health Survey

Background:  To determine access to and utilization of eye health services for indigenous Australians. Design:  A national, stratified, random cluster sample was drawn from 30 communities across Australia that each included about 300 indigenous people. Participants:  A total of 1189 indigenous adults aged 40 and above were examined, representing 79% of the target population. Methods:  Eye health services data including nature and availability of facilities and workforce supply were collected for comparison with eye health prevalence data. The data were collected in 2008. Main Outcome Measures:  Low vision prevalence and coverage rate for distance refractive correction. Results:  The full-time equivalent availability of an optometrist working in an Aboriginal Medical Service was significantly associated with both a decrease in the prevalence of low vision (t = -2.41, P = 0.02) and an increase in the coverage rate for distance refractive correction (t = 2.99, P = 0.006). These associations were not replicated when comparing availability of private or hospital-based optometry in each community. Regional eye health coordinators appeared to provide an improved utilization of Aboriginal Health Services and therefore improved access to Aboriginal medical service optometry. Conclusions:  Eye health services for indigenous Australians need to be provided in culturally appropriate facilities with clear links to the indigenous community to optimize access to care and reduce the prevalence of vision impairment. The adequate provision of accessible eye care services is an important component in 'closing the gap' in vision loss for indigenous Australians.

Factors Associated with Urge Urinary Incontinence After Surgery for Stress Urinary Incontinence

: The objectives of the study were to estimate the severity of urge urinary incontinence (UUI) after surgery for stress incontinence and identify factors associated with symptom severity.

Genetic Variation in Eggshell Crystal Size and Orientation is Large and These Traits Are Correlated with Shell Thickness and Are Associated with Eggshell Matrix Protein Markers

The size and orientation of calcium carbonate crystals influence the structure and strength of the eggshells of chickens. In this study, estimates of heritability were found to be high (0.6) for crystal size and moderate (0.3) for crystal orientation. There was a strong positive correlation (0.65) for crystal size and orientation with the thickness of the shell and, in particular, with the thickness of the mammillary layer. Correlations with shell breaking strength were positive but with a high standard error. This was contrary to expectations, as in man-made materials smaller crystals would be stronger. We believe the results of this study support the hypothesis that the structural organization of shell, and in particular the mammillary layer, is influenced by crystal size and orientation, especially during the initial phase of calcification. Genetic associations for crystal measurements were observed between haplotype blocks or individual markers for a number of eggshell matrix proteins. Ovalbumin and ovotransferrin (LTF) markers for example were associated with crystal size, while ovocleidin-116 and ovocalyxin-32 (RARRES1) markers were associated with crystal orientation. The location of these proteins in the eggshell is consistent with different phases of the shell-formation process. In conclusion, the variability of crystal size, and to a lesser extent orientation, appears to have a large genetic component, and the formation of calcite crystals are intimately related to the ultrastructure of the eggshell. Moreover, this study also provides evidence that proteins in the shell influence the variability of crystal traits and, in turn, the shell's thickness profile. The crystal measurements and/or the associated genetic markers may therefore prove to be useful in selection programs to improve eggshell quality.

Delivery of Multiple SiRNAs Using Lipid-coated PLGA Nanoparticles for Treatment of Prostate Cancer

Nanotechnology can provide a critical advantage in developing strategies for cancer management and treatment by helping to improve the safety and efficacy of novel therapeutic delivery vehicles. This paper reports the fabrication of poly(lactic acid-co-glycolic acid)/siRNA nanoparticles coated with lipids for use as prostate cancer therapeutics made via a unique soft lithography particle molding process called Particle Replication In Nonwetting Templates (PRINT). The PRINT process enables high encapsulation efficiency of siRNA into neutral and monodisperse PLGA particles (32-46% encapsulation efficiency). Lipid-coated PLGA/siRNA PRINT particles were used to deliver therapeutic siRNA in vitro to knockdown genes relevant to prostate cancer.

Optimization of a Novel Kinase Inhibitor Scaffold for the Dual Inhibition of JAK2 and FAK Kinases

The elaboration of a novel scaffold for the inhibition of JAK2 and FAK kinases was targeted in order to provide a dual inhibitor that could target divergent pathways for tumor cell progression.

Staph ID/R: a Rapid Method for Determining Staphylococcus Species Identity and Detecting the MecA Gene Directly from Positive Blood Culture

Rapid diagnosis of staphylococcal bacteremia directs appropriate antimicrobial therapy, leading to improved patient outcome. We describe herein a rapid test (<75 min) that can identify the major pathogenic strains of Staphylococcus to the species level as well as the presence or absence of the methicillin resistance determinant gene, mecA. The test, Staph ID/R, combines a rapid isothermal nucleic acid amplification method, helicase-dependent amplification (HDA), with a chip-based array that produces unambiguous visible results. The analytic sensitivity was 1 CFU per reaction for the mecA gene and was 1 to 250 CFU per reaction depending on the staphylococcal species present in the positive blood culture. Staph ID/R has excellent specificity as well, with no cross-reactivity observed. We validated the performance of Staph ID/R by testing 104 frozen clinical positive blood cultures and comparing the results with rpoB gene or 16S rRNA gene sequencing for species identity determinations and mecA gene PCR to confirm mecA gene results. Staph ID/R agreed with mecA gene PCR for all samples and agreed with rpoB/16S rRNA gene sequencing in all cases except for one sample that contained a mixture of two staphylococcal species, one of which Staph ID/R correctly identified, for an overall agreement of 99.0% (P < 0.01). Staph ID/R could potentially be used to positively affect patient management for Staphylococcus-mediated bacteremia.

Nonmedical Factors Associated with Prolonged Hospital Length of Stay in an Urban Homebound Population

Prolonged length of stay (LOS) is a major concern for hospitalized populations at risk for adverse events. Homebound patients are at particular risk for long stays and may have unique discharge needs because of their commitment to be cared for at home despite poor functional status.

Modeling the Activation of Tobacco Smoking Expectancies in Memory in Relation to Use Patterns

Methodology that has led to successful strategies to reduce alcohol use was applied to tobacco smoking expectancies. Individual differences scaling was used to empirically model a semantic network of associations stored in memory and preference mapping was used to model likely paths of expectancy activation for groups with different smoking histories. Smokers emphasized an external appearance-internal experience dimension and were more likely to activate expectancies of negative affect reduction. Nonsmokers emphasized a positive-negative dimension and were more likely to activate expectancies of health risks and reduced physical attractiveness. Proportionate frequencies of first associates' validated findings of the MDS-based solutions. Future efforts to alter likely activation patterns may successfully reduce the onset of smoking, enhance quit rates, and reduce relapse.

Extraordinary Transgressive Phenotypes of Hybrid Tomato Are Influenced by Epigenetics and Small Silencing RNAs

Hybrid organisms may fail to develop, be sterile or they may be more vigorous than either of the parents. Examples of hybrid vigour or hybrid necrosis in the F1 are often not inherited stably in subsequent generations if they are associated with overdominance. There can also be transgressive phenotypes that are inherited stably in these later generations, but the underlying mechanisms are not well understood. Here we have investigated the possibility that stable transgressive phenotypes in the progeny of crosses between cultivated tomato (Solanum lycopersicum cv. M82) and a wild relative (Solanum pennellii, accession LA716) are associated with micro or small interfering(si) RNAs. We identified loci from which these small(s)RNAs were more abundant in hybrids than in either parent and we show that accumulation of such transgressive sRNAs correlated with suppression of the corresponding target genes. In one instance this effect was associated with hypermethylation of the corresponding genomic DNA. Our results illustrate a potential role of transgressive sRNAs in plant breeding and in natural evolution with wild plants.

Green Tea Polyphenols and 1-α-OH-vitamin D₃ Attenuate Chronic Inflammation-induced Myocardial Fibrosis in Female Rats

Studies have suggested that 1-α-OH-vitamin D₃ and green tea polyphenols (GTPs) are promising dietary supplements for mitigating chronic inflammation-induced fibrosis of vessels because of their anti-inflammatory properties. This study evaluated (1) the impact of 1-α-OH-vitamin D₃ on myocardial fibrosis in female rats with chronic inflammation and (2) if 1-α-OH-vitamin D₃ and GTPs have an additive or synergistic effect to attenuate myocardial fibrosis in these female rats. A 3-month study of a 2 (no 1-α-OH-vitamin D₃ vs. 0.05 μg/kg 1-α-OH-vitamin D₃, five times per week) ×2 (no GTPs vs. 0.5% GTPs in drinking water) factorial design in lipopolysaccharide (LPS)-administered female rats was performed. Additionally, a group receiving placebo administration was used to compare with a group receiving LPS administration only to evaluate the effect of LPS. Masson's Trichrome staining evaluated myocardial fibrosis in coronary vessels and surrounding myocardium. Spleen cyclooxygenase-2 mRNA expression was determined by real-time polymerase chain reaction. Total lipid profiles were also determined. Whole blood was used for differential cell counts. Data were analyzed by two-way analysis of variance followed by mean separation procedures. At 3 months LPS administration induced myocardial fibrosis in vessels and surrounding myocardium, spleen cyclooxygenase-2 mRNA expression, and elevated leukocyte counts, whereas both 1-α-OH-vitamin D₃ administration and GTPs supplementation significantly attenuated these pro-inflammatory events. The inhibitory effects of 1-α-OH-vitamin D₃ and GTPs seem to be an individual effect, instead of an additive or synergistic effect. 1-α-OH-vitamin D₃ and GTPs lowered red blood cell counts, hematocrit, and hemoglobin. Neither 1-α-OH-vitamin D₃ nor GTPs affected lipid profiles. In summary, both 1-α-OH-vitamin D₃ administration and GTPs supplementation mitigate myocardial fibrosis through suppression of a chronic inflammation innate immune response.

Autophagy in the Retina: a Potential Role in Age-related Macular Degeneration

Strain Tunes Proteolytic Degradation and Diffusive Transport in Fibrin Networks

Proteolytic degradation of fibrin, the major structural component in blood clots, is critical both during normal wound healing and in the treatment of ischemic stroke and myocardial infarction. Fibrin-containing clots experience substantial strain due to platelet contraction, fluid shear, and mechanical stress at the wound site. However, little is understood about how mechanical forces may influence fibrin dissolution. We used video microscopy to image strained fibrin clots as they were degraded by plasmin, a major fibrinolytic enzyme. Applied strain causes up to 10-fold reduction in the rate of fibrin degradation. Analysis of our data supports a quantitative model in which the decrease in fibrin proteolysis rates with strain stems from slower transport of plasmin into the clot. We performed fluorescence recovery after photobleaching (FRAP) measurements to further probe the effect of strain on diffusive transport. We find that diffusivity perpendicular to the strain axis decreases with increasing strain, while diffusivity along the strain axis remains unchanged. Our results suggest that the properties of the fibrin network have evolved to protect mechanically loaded fibrin from degradation, consistent with its function in wound healing. The pronounced effect of strain upon diffusivity and proteolytic susceptibility within fibrin networks offers a potentially useful means of guiding cell growth and morphology in fibrin-based biomaterials.

The Effect of Weather on Walking Behavior in Older Adults

In this article, the authors examine how temperature and precipitation affect the probability that a retired American between the ages of 65 and 90 walks at least 2.5 hr/wk, using longitudinal data on walking frequency from the Consumption and Activities Mail Survey, a subpanel in the Health and Retirement Survey. Walking behavior is linked with monthly temperature and precipitation data from weather-station reports. The authors found that higher temperatures were associated with a higher probability of walking at least 2.5 hr/wk for women. In contrast, higher temperatures are associated with a lower probability of walking at least 2.5 hr/wk among men. Precipitation is not significantly associated with walking behavior for either gender.

Resolution of Extra-axial Collections After Decompressive Craniectomy for Ischemic Stroke

Extra-axial fluid collections are known consequences of decompressive hemicraniectomy. Studies have examined these collections and their management. We retrospectively reviewed 12 consecutive patients who underwent decompressive hemicraniectomy for the treatment of malignant cerebral edema after infarction and evaluated the evolution, resolution and treatment of post-operative extra-axial fluid collections. All patients underwent standard-sized frontotemporoparietal hemicraniectomy with duraplasty as treatment for medically intractable malignant cerebral edema at an average of 3 days after the stroke (median 2 days). Their 30-day mortality was 25%. Three patients developed some extra-axial fluid collections after craniectomy: two patients developed the collections early in their post-operative course, 3 days and 5 days after the craniectomy. Both experienced spontaneous resolution of the collections without corrective cranioplasty or shunt placement at 34 days and 58 days after surgery. The third patient developed a collection 55 days after the operation related to a subgaleal bacterial infection. In the final analysis, 18% of patients developed extra-axial collections and all resolved spontaneously. The incidence of extra-axial collections after decompressive hemicraniectomy following ischemic stroke was lower in our retrospective series than has been reported by others. The collections resolved spontaneously, suggesting that early anticipatory, corrective treatment with cerebrospinal fluid diversion or cranioplasty may not be warranted.

Modelling the Role of the Basement Membrane Beneath a Growing Epithelial Monolayer

The role of the basement membrane is vital in maintaining the integrity and structure of an epithelial layer, acting as both a mechanical support and forming the physical interface between epithelial cells and the surrounding connective tissue. The function of this membrane is explored here in the context of a growing epithelial monolayer, defined such that the epithelial cells divide and migrate along a deformable substrate. A discrete, off-lattice cell-centre modelling approach is undertaken, which permits definition of a basement membrane component, separating the epithelial cells from the tissue stroma whilst responding to forces from both that arise due to cell division, migration and apoptosis. This model is applicable to a range of biological epithelia, including the self-renewing interfollicular epidermis, the olfactory epithelium and the intestinal crypts of Lieberkühn, to inform response and recovery of such tissues following injury. Model simulations show that homeostasis of the growing monolayer can be achieved and sustained, and the necessary balance of interactive cell forces, cell migration and cell death is presented. This work is proposed as a novel extension to the body of discrete models of biological epithelia, permitting investigation of the growth and migration of epithelial cells in a deformable environment.

Liver Transplantation in the Management of Unresectable Hepatoblastoma in Children

Complete surgical resection is essential to long-term survival in children with hepatoblastoma. We present the guidelines from the Children's Oncology Group (COG), liver tumor study group of the Societe Internationale Oncologie Pediatrique (SIOPEL), and German Pediatric Oncology Group (GPOH) for early referral of children with potentially unresectable hepatoblastoma to a specialty center with expertise in extreme resection and liver transplantation. Patients who will become candidates for liver transplantation should receive chemotherapy following the same protocols as for children undergoing a partial hepatectomy. The Pediatric Liver Unresectable Tumor Observatory (PLUTO) is an international prospective database established to collect data and make future recommendations on controversial issues regarding the use of transplant in hepatoblastoma including: 1) What is the optimal treatment of multifocal tumors. 2) What is the role of extreme resection vs. liver transplant in patients with major venous involvement. 3) What is the role of transplant in patients who present with lung metastasis. 3) Should patients with tumor relapse be offered a rescue transplant. 4) What is the role of pre- and post- transplant chemotherapy.

Molecular Diagnosis of Subcutaneous Pythium Insidiosum Infection by Use of PCR Screening and DNA Sequencing

Pythium insidiosum is an emerging human pathogen classified among brown algae and diatoms that can cause significant morbidity and mortality in otherwise healthy individuals. Here we describe a pediatric patient with pythiosis acquired in the southern United States, diagnosed by molecular screening and DNA sequencing of internal transcribed spacer region 1.

Whole-genome Sequences of Borrelia Bissettii, Borrelia Valaisiana, and Borrelia Spielmanii

It has been known for decades that human Lyme disease is caused by the three spirochete species Borrelia burgdorferi, Borrelia afzelii, and Borrelia garinii. Recently, Borrelia valaisiana, Borrelia spielmanii, and Borrelia bissettii have been associated with Lyme disease. We report the complete genome sequences of B. valaisiana VS116, B. spielmanii A14S, and B. bissettii DN127.

Delivery of VEGF Using Collagen-coated Polycaprolactone Scaffolds Stimulates Angiogenesis

Establishing sufficient vascularization in scaffold remains a challenge for tissue-engineering. To improve angiogenesis, we incorporated vascular endothelial growth factor (VEGF) in collagen-coating over the porous polycaprolactone (PCL) scaffolds. The release kinetics of loaded VEGF from collagen-coated PCL (col-PCL) scaffolds was same as from scaffolds without the collagen. The bioactivity of VEGF delivered by the col-PCL scaffolds was confirmed by human umbilical vein endothelial cell (HUVEC) proliferation and chorioallantoic membrane (CAM) assay. The col-PCL scaffolds were implanted subcutaneously in mice for 7 and 14 days. At day 7, vascularization within scaffolds loaded with VEGF was superior to that in the scaffolds without VEGF. However, the vessel connectivity to host circulatory system was incomplete and restricted to the scaffold edges. At day 14, blood vessels in scaffolds reached density similar to the subcutaneous tissue and were perfusable throughout the implant thickness. Prewashing the scaffolds with saline to remove the unbound growth factor decreased the initial burst release and sustained the VEGF-mediated angiogenesis in vivo. In conclusion, our study demonstrates that physically adsorbed VEGF stimulated angiogenesis in collagen-coated PCL scaffolds.

Medical Education in the Clouds

Ovariohysterectomy Versus Ovariectomy

Clinical Bleeding Events and Laboratory Coagulation Profiles in Acute Promyelocytic Leukemia

Bleeding is the leading cause of death for patients with acute promyelocytic leukemia (APL). Blood component transfusion to correct coagulopathy is the keystone in reducing bleeding. The benefit of fresh frozen plasma transfusion is unproven. Using laboratory profiles to predict bleeding is important guidance for the determination of transfusion policies in the treatment of APL.

The Role of Relationship Quality and Perceived Partner Responses with Pain and Disability in Those with Back Pain

The objectives of this study were to investigate the associations of key constructs of relationship quality (cohesion, consensus, and satisfaction) and perceived partner responses to pain behavior (e.g., solicitous and negative responses) with the outcomes of pain and disability in those with long-term low back pain, and to explore the role of the patient's depressive symptom mood state on those associations.

The Field of Proteomics Continues to Expand

Coping with Chronic Illness in Childhood and Adolescence

Chronic illnesses and medical conditions present millions of children and adolescents with significant stress that is associated with risk for emotional and behavioral problems and interferes with adherence to treatment regimens. We review research on the role of child and adolescent coping with stress as an important feature of the process of adaptation to illness. Recent findings support a control-based model of coping that includes primary control or active coping (efforts to act on the source of stress or one's emotions), secondary control or accommodative coping (efforts to adapt to the source of stress), and disengagement or passive coping (efforts to avoid or deny the stressor). Evidence suggests the efficacy of secondary control coping in successful adaptation to chronic illness in children and adolescents, disengagement coping is associated with poorer adjustment, and findings for primary control coping are mixed. Avenues for future research are highlighted.

Prevalence of Malignancies Among U.S. Male Patients with Haemophilia: a Review of the Haemophilia Surveillance System

Summary.  The prevalence of malignancies in US male patients with haemophilia, with or without concomitant viral infections, remains unknown. To estimate the prevalence of malignancy in US male patients with haemophilia. We investigated the prevalence of malignancies among male patients with haemophilia using data from a six-state haemophilia surveillance project. Case patients with malignancies were identified using International Classification of Diseases, 9th Revision, Clinical Modification codes abstracted from hospital records and death certificates during the surveillance period. Cancer prevalence rates were calculated for each year during the surveillance and compared with age- and race-specific prevalence rates among the U.S. male population obtained from the Surveillance, Epidemiology and End Results (SEER) Program. A total of 7 cases of leukaemia, 23 cases of lymphoma and 56 classifiable solid malignancies were identified among 3510 case patients during a total of 15 330 annual data abstraction collections. The rates of leukaemia, lymphoma and liver cancer among case patients were significantly higher than the rates among U.S. males as judged by prevalence ratios of 3.1 [95% confidence interval (CI) = 1.4-7.0] and 2.9 (95% CI = 1.8-4.6), respectively. In contrast, the prevalence ratio of prostate cancer was lower than expected at 0.49 (95% CI = 0.31-0.77). Overall the prevalence of most cancers among case patients was similar to that of the U.S. male population. However, patients with haemophilia who have unexplained symptoms should be evaluated for malignancy.

Duration of Red Blood Cell Storage and Outcomes Following Orthotopic Liver Transplantation

Liver transplantation may be complicated by massive intraoperative bleeding, and red blood cell (RBC) transfusions may be required. The storage duration or age of transfused RBCs has been shown to affect the morbidity and mortality of critically ill, trauma, and cardiac surgery patients. Here we investigate the effect of RBC age on the outcomes of liver transplant patients. Five hundred thirty-one patients underwent orthotopic liver transplantation between January 1, 2000 and August 15, 2010. The patient demographics, the Model for End-Stage Liver Disease-sodium (MELD-Na) score, and the number and age of RBC units were evaluated with univariate and multivariate models of outcomes, which included mortality rates 2 years after transplantation, postoperative infections, and organ rejection. In a univariate analysis, the number of RBC units (but not the RBC age) was associated with increased 2-year mortality, an increased risk of infection, and a decreased risk of organ rejection. Only the number of RBC units was associated with increased 2-year mortality in a multivariate Cox regression model. The mortality risk was decreased by two-thirds for patients who received <10 U of RBCs versus those who received ≥10 U (hazard ratio = 0.33, 95% confidence interval = 0.16-0.69, P = 0.003). The number of transfused RBC units was not associated with the risk of infection or organ rejection in a multivariate logistic regression model. In conclusion, the RBC age is not associated with infection, organ rejection, or death in liver transplant patients. Patients who receive more blood have an increased risk of death. In a multivariate model, the MELD-Na score was not associated with increased mortality, and this is consistent with previous studies demonstrating that the MELD-Na score is a poor predictor of long-term survival after transplantation. Liver Transpl 18:475-481, 2012. © 2012 AASLD.

Development of an Endoluminal Intestinal Lengthening Capsule

Prior studies demonstrated the ability of a spring to lengthen intestinal segments. We made two innovations to this device. First, we employed a degradable capsule to control the deployment of the spring. Second, we decreased the spring force to allow slower expansion of the intestinal segment.

Engraftment of Human HSCs in Nonirradiated Newborn NOD-scid IL2rγnull Mice is Enhanced by Transgenic Expression of Membrane-bound Human SCF

Immunodeficient mice engrafted with human HSCs support multidisciplinary translational experimentation, including the study of human hematopoiesis. Heightened levels of human HSC engraftment are observed in immunodeficient mice expressing mutations in the IL2-receptor common γ chain (IL2rg) gene, including NOD-scid IL2rγ(null) (NSG) mice. Engraftment of human HSC requires preconditioning of immunodeficient recipients, usually with irradiation. Such preconditioning increases the expression of stem cell factor (SCF), which is critical for HSC engraftment, proliferation, and survival. We hypothesized that transgenic expression of human membrane-bound stem cell factor Tg(hu-mSCF)] would increase levels of human HSC engraftment in nonirradiated NSG mice and eliminate complications associated with irradiation. Surprisingly, detectable levels of human CD45(+) cell chimerism were observed after transplantation of cord blood-derived human HSCs into nonirradiated adult as well as newborn NSG mice. However, transgenic expression of human mSCF enabled heightened levels of human hematopoietic cell chimerism in the absence of irradiation. Moreover, nonirradiated NSG-Tg(hu-mSCF) mice engrafted as newborns with human HSCs rejected human skin grafts from a histoincompatible donor, indicating the development of a functional human immune system. These data provide a new immunodeficient mouse model that does not require irradiation preconditioning for human HSC engraftment and immune system development.

Combined Two-photon Luminescence Microscopy and OCT for Macrophage Detection in the Hypercholesterolemic Rabbit Aorta Using Plasmonic Gold Nanorose

The macrophage is an important early cellular marker related to risk of future rupture of atherosclerotic plaques. Two-channel two-photon luminescence (TPL) microscopy combined with optical coherence tomography (OCT) was used to detect, and further characterize the distribution of aorta-based macrophages using plasmonic gold nanorose as an imaging contrast agent.

Preferences for Rapid Point-of-Care HIV Testing in Primary Care

Although the identification of individuals infected with HIV is an important element of treatment and prevention programs, many people living with HIV are unaware of their status. Thus, individuals are unable to benefit from treatment, and preventable HIV transmission continues to occur. Rapid point-of-care testing for HIV has been found to be preferred by patients in some contexts. However, few studies have examined preferences in primary care populations. This study investigates HIV testing preferences within an urban primary care clinic. Employing a cross-sectional design, data were collected on demographic characteristics, HIV risk factors, and testing history and preferences of participants. A total of 81% of participants stated that they would prefer rapid testing to standard testing, a finding that is consistent across demographic variables and risk factors examined. Increased availability of this modality may decrease barriers to HIV testing, with positive implications both for clinical management of HIV infection and prevention of HIV transmission.

Associations Among Visual Acuity and Vision- and Health-related Quality of Life Among Patients in the Multicenter Uveitis Steroid Treatment Trial

To evaluate the associations between visual acuity and self-reported visual function; visual acuity and health-related quality of life (QoL) metrics; a summary measure of self-reported visual function and health-related QoL; and individual domains of self-reported visual function and health-related QoL in patients with uveitis.

MKNK1 is a YB-1 Target Gene Responsible for Imparting Trastuzumab Resistance and Can Be Blocked by RSK Inhibition

Trastuzumab (Herceptin) resistance is a major obstacle in the treatment of patients with HER2-positive breast cancers. We recently reported that the transcription factor Y-box binding protein-1 (YB-1) leads to acquisition of resistance to trastuzumab in a phosphorylation-dependent manner that relies on p90 ribosomal S6 kinase (RSK). To explore how this may occur we compared YB-1 target genes between trastuzumab-sensitive cells (BT474) and those with acquired resistance (HR5 and HR6) using genome-wide chromatin immunoprecipitation sequencing (ChIP-sequencing), which identified 1391 genes uniquely bound by YB-1 in the resistant cell lines. We then examined differences in protein expression and phosphorylation between these cell lines using the Kinexus Kinex antibody microarrays. Cross-referencing these two data sets identified the mitogen-activated protein kinase-interacting kinase (MNK) family as potentially being involved in acquired resistance downstream from YB-1. MNK1 and MNK2 were subsequently shown to be overexpressed in the resistant cell lines; however, only the former was a YB-1 target based on ChIP-PCR and small interfering RNA (siRNA) studies. Importantly, loss of MNK1 expression using siRNA enhanced sensitivity to trastuzumab. Further, MNK1 overexpression was sufficient to confer resistance to trastuzumab in cells that were previously sensitive. We then developed a de novo model of acquired resistance by exposing BT474 cells to trastuzumab for 60 days (BT474LT). Similar to the HR5/HR6 cells, the BT474LT cells had elevated MNK1 levels and were dependent on it for survival. In addition, we demonstrated that RSK phosphorylated MNK1, and that this phosphorylation was required for ability of MNK1 to mediate resistance to trastuzumab. Furthermore, inhibition of RSK with the small molecule BI-D1870 repressed the MNK1-mediated trastuzumab resistance. In conclusion, this unbiased integrated approach identified MNK1 as a player in mediating trastuzumab resistance as a consequence of YB-1 activation, and demonstrated RSK inhibition as a means to overcome recalcitrance to trastuzumab.Oncogene advance online publication, 16 January 2012. doi:10.1038/onc.2011.617.

Clinical and Pathological Heterogeneity of Cervical Intraepithelial Neoplasia Grade 3

Cervical intraepithelial neoplasia grade 3 (CIN3), the immediate cervical cancer precursor, is a target of cervical cancer prevention. However, less than half of CIN3s will progress to cancer. Routine treatment of all CIN3s and the majority of CIN2s may lead to overtreatment of many lesions that would not progress. To improve our understanding of CIN3 natural history, we performed a detailed characterization of CIN3 heterogeneity in a large referral population in the US.

Osteosarcoma at the Site of Titanium Orthopaedic Implants in a Dog

An Alaskan Malamute underwent unilateral tibial tuberosity advancement (TTA) surgery to stabilise a stifle joint with a deficient cranial cruciate ligament. The dog made an excellent recovery with no postoperative complications, until 20 months post-surgery when he presented with acute onset ipsilateral pelvic limb lameness. Osteosarcoma (OSA) was diagnosed adjacent to the titanium implants. Currently, there is a paucity of information on the epidemiology of OSA adjacent to orthopaedic implants in canine patients. The clinical, radiological and pathological findings of this case of periprosthetic OSA, and a potential causal relationship between titanium implants and bone neoplasia, are discussed.

Frequency and Patterns of Protease Gene Resistance Mutations in HIV-infected Patients Treated with Lopinavir/ritonavir As Their First Protease Inhibitor

Selection of protease mutations on antiretroviral therapy (ART) including a ritonavir-boosted protease inhibitor (PI) has been reported infrequently. Scarce data exist from long-term cohorts on resistance incidence or mutational patterns emerging to different PIs.

Clinical Course, Characteristics and Prognostic Indicators in Patients Presenting with Back and Leg Pain in Primary Care. The ATLAS Study Protocol

Low-back related leg pain with or without nerve root involvement is associated with a poor prognosis compared to low back pain (LBP) alone. Compared to the literature investigating prognostic indicators of outcome for LBP, there is limited evidence on prognostic factors for low back-related leg pain including the group with nerve root pain. This 1 year prospective consultation-based observational cohort study will describe the clinical, imaging, demographic characteristics and health economic outcomes for the whole cohort, will investigate differences and identify prognostic indicators of outcome (i.e. change in disability at 12 months), for the whole cohort and, separately, for those classified with and without nerve root pain. In addition, nested qualitative studies will provide insights on the clinical consultation and the impact of diagnosis and treatment on patients' symptom management and illness trajectory.

Wake-promoting Agents: Search for Next Generation Modafinil: Part I

In search of a next generation molecule to the novel wake promoting agent modafinil, a series of bi-phenyl derived wakefulness enhancing agents (in rat) was developed. From this work, compound 17 has been selected for additional studies.

Clinical Outcomes in Patients with Cytomegalovirus Retinitis Treated with Ganciclovir Implant

To describe the clinical outcomes of patients with cytomegalovirus (CMV) retinitis and AIDS treated with ganciclovir implant.

Repeated Oral Administration of Lipopolysaccharide from Escherichia Coli 0111:B4 Modulated Humoral Immune Responses in Periparturient Dairy Cows

The objective of this study was to evaluate the effects of repeated oral exposure to LPS on humoral immune responses of periparturient dairy cows. Sixteen Holstein cows were assigned to two treatment groups 2 wk before the expected day of parturition. Cows were administered orally, twice weekly at wk -2, -1 and +1 around parturition, with the following treatments: 3 ml saline; or 3 ml of saline containing LPS from Escherichia coli 0111:B4. The amount of LPS administered during wk -2, -1, and +1 was 0.01, 0.05, or 0.1 µg/kg body weight, respectively. Multiple blood samples were collected by jugular vein and various immune and clinical variables were measured. Results indicated that, on one hand, concentrations of plasma IgG anti-LPS Abs decreased (P < 0.01) and those of IgM anti-LPS Abs increased (P < 0.01) in cows treated with oral LPS. On the other hand, there were no overall differences (P > 0.05) in the concentrations of serum amyloid A, LPS-binding protein, haptoglobin, cortisol, IgA anti-LPS Abs in the plasma, feed intake, body temperature and rumen contractions rate between the control and treatment groups. To our knowledge, this study is the first to show that repeated oral administration with LPS from E. coli 0111:B4 has the potential to stimulate humoral immune responses in periparturient dairy cows.

Topical Antimicrobial Treatment of Acne Vulgaris: an Evidence-based Review

Topical antimicrobial treatment is indicated for mild to moderate acne vulgaris. Our literature review includes searches of Ovid, MEDLINE, EMBASE, and the databases of the Cochrane Library. A detailed search strategy is included. All searches were limited to controlled trials and systematic reviews. No year limits were applied to the searches, but we focused on trials, guidelines, and reviews published since 2004, the year that the last review of topical antimicrobials was published in this journal. Several controlled trials demonstrate that benzoyl peroxide, topical antibiotics, and topical retinoids used in combination provide the greatest efficacy and safety profile for the treatment of mild to moderate acne, but there are few trials directly comparing different combinations of these topical therapies with one another. Additionally, robust studies comparing cost and efficacy of generic combinations of the above agents with proprietary fixed-dose combination therapies that may increase compliance are also lacking. Although they have not been extensively studied, alternative agents including dapsone, salicylic acid, azelaic acid, and zinc are safe and efficacious when combined with traditional therapies.

Affective Variability Predicts Suicidal Ideation in Individuals at Ultra-high Risk of Developing Psychosis: an Experience Sampling Study

There is a suggestion in the literature that more variable affect increases suicidal ideation through the repeated re-activation of latent suicidal cognitions. The hypothesis that affective variability would be a better predictor of suicidal ideation and related behaviour than affect level was tested in individuals at ultra-high risk of developing psychosis. This study also examined the prediction that affective variability is a suicide-specific mechanism and would not predict levels of attenuated psychotic phenomena.

Open Disclosure of Adverse Events: Transparency and Safety in Health Care

Many patients suffering adverse events in health care turn to the legal system to learn what happened to them and to seek compensation. Health care providers have ethical, professional, and legal duties to disclose the harmful effects of care to the patient, regardless of how small the risk. The purpose of open disclosure is to explain what happened to the patient and to seek a just outcome for patient and provider. This article explores our experience of managing and implementing an open disclosure program in an acute and chronic tertiary care facility with university affiliation in the Veterans Health Administration.

A High Calcium Diet Containing Nonfat Dry Milk Reduces Weight Gain and Associated Adipose Tissue Inflammation in Diet-induced Obese Mice when Compared to High Calcium Alone

ABSTRACT:

Annual Mammography Screening: is It Necessary?

Recent recommendations from the U.S. Preventative Services Task Force suggest that screening mammography for women should be biennial starting at age 50 years and continue to age 74 years. With these recommendations in mind, we proposed a study to evaluate women at our institution in whom breast cancer is diagnosed within 1 year of a previously benign mammogram. A retrospective chart review was performed over a 4-year period. Only patients who had both diagnostic mammograms and previous mammograms performed at our institution and a pathologic diagnosis of breast cancer were included. Benign mammograms were defined as either Breast Imaging Reporting And Data System 1 or 2. Analysis of the time elapse between benign mammogram and subsequent mammogram indicative of the diagnosis of breast cancer was performed. A total of 205 patients were included. The average age was 64 years. From our results, 48 patients, 23 per cent of the total, had a documented benign mammogram at 12 months or less before a breast cancer diagnosis. One hundred forty-three (70%) patients had a benign mammogram at 18 months or less prior. This study raises concern that 2 years between screening mammograms may delay diagnosis and possible treatment options for many women.

Does Discordancy Between the CD4 Count and CD4 Percentage in HIV-positive Individuals Influence Outcomes on Highly Active Antiretroviral Therapy?

The CD4 count and CD4 percentage (CD4%) are both strong predictors of clinical disease progression in human immunodeficiency virus (HIV). Although individuals may show discordancy between their CD4 count and CD4%, the clinical relevance of this is unclear.

Atomic Inner-shell X-ray Laser at 1.46 Nanometres Pumped by an X-ray Free-electron Laser

Since the invention of the laser more than 50 years ago, scientists have striven to achieve amplification on atomic transitions of increasingly shorter wavelength. The introduction of X-ray free-electron lasers makes it possible to pump new atomic X-ray lasers with ultrashort pulse duration, extreme spectral brightness and full temporal coherence. Here we describe the implementation of an X-ray laser in the kiloelectronvolt energy regime, based on atomic population inversion and driven by rapid K-shell photo-ionization using pulses from an X-ray free-electron laser. We established a population inversion of the Kα transition in singly ionized neon at 1.46 nanometres (corresponding to a photon energy of 849 electronvolts) in an elongated plasma column created by irradiation of a gas medium. We observed strong amplified spontaneous emission from the end of the excited plasma. This resulted in femtosecond-duration, high-intensity X-ray pulses of much shorter wavelength and greater brilliance than achieved with previous atomic X-ray lasers. Moreover, this scheme provides greatly increased wavelength stability, monochromaticity and improved temporal coherence by comparison with present-day X-ray free-electron lasers. The atomic X-ray lasers realized here may be useful for high-resolution spectroscopy and nonlinear X-ray studies.

Ten-year Outcome After Rapid Discontinuation of Prednisone in Adult Primary Kidney Transplantation

Rapid discontinuation of prednisone after kidney transplantation potentially allows for minimization of steroid-related side effects. Although intermediate-term data with rapid discontinuation of prednisone have been promising, concern still exists regarding long-term outcomes. The 10-year experience is reported herein.

Identification of Distinct Subgroups of Breast Cancer Patients Based on Self-reported Changes in Sleep Disturbance

PURPOSE: The purposes of this study were to identify distinct subgroups of patients based on self-reported sleep disturbance prior to through 6 months after breast cancer surgery and evaluate for differences in demographic, clinical, and symptom characteristics among these latent classes. METHODS: Women (n = 398) who underwent unilateral breast cancer surgery were enrolled prior to surgery. Patients completed measures of functional status, sleep disturbance (i.e., General Sleep Disturbance Scale (GSDS); higher scores indicate higher levels of sleep disturbance), fatigue, attentional fatigue, depressive symptoms, and anxiety prior to surgery and monthly for 6 months. RESULTS: Three distinct classes of sleep disturbance trajectories were identified using growth mixture modeling. The high sustained class (55.0%) had high and the low sustained class (39.7%) had low GSDS scores prior to surgery that persisted for 6 months. The decreasing class (5.3%) had high GSDS score prior to surgery that decreased over time. Women in the high sustained class were significantly younger, had more comorbidity and poorer function, and were more likely to report hot flashes compared to the low sustained class. More women who underwent mastectomy or breast reconstruction were in the decreasing class. Decreasing and high sustained classes reported higher levels of physical fatigue, attentional fatigue, depressive symptoms, and anxiety compared to the low sustained class. CONCLUSIONS: A high percentage of women has significant sleep disturbance prior to surgery that persists during subsequent treatments (i.e., radiation therapy and chemotherapy). Clinicians need to perform routine assessments and initiate appropriate interventions to improve sleep prior to and following surgery.

Boosting the Globalization of Plant Proteomics Through INPPO: Current Developments and Future Prospects

The International Plant Proteomics Organization (INPPO) is a non-profit-organization consisting of people who are involved or interested in plant proteomics. INPPO is constantly growing in volume and activity, which is mostly due to the realization among plant proteomics researchers worldwide for the need of such a global platform. Their active participation resulted in the rapid growth within the first year of INPPO's official launch in 2011 via its website (www.inppo.com) and publication of the 'Viewpoint paper' in a special issue of PROTEOMICS (May 2011). Here, we will be highlighting the progress achieved in the year 2011 and the future targets for the year 2012 and onwards. INPPO has achieved a successful administrative structure, the Core Committee (CC; composed of President, Vice-President, and General Secretaries), Executive Council (EC), and General Body (GB) to achieve INPPO objectives. Various committees and subcommittees are in the process of being functionalized via discussion amongst scientists around the globe. INPPO's primary aim to popularize the plant proteomics research in biological sciences has also been recognized by PROTEOMICS where a section dedicated to plant proteomics has been introduced starting January 2012, following the very first issue of this journal devoted to plant proteomics in May 2011. To disseminate organizational activities to the scientific community, INPPO has launched a biannual (in January and July) newsletter entitled 'INPPO Express: News & Views' with the first issue published in January 2012. INPPO is also planning to have several activities in 2012, including programs within the Education Outreach committee in different countries, and the development of research ideas and proposals with priority on crop and horticultural plants, while keeping tight interactions with proteomics programs on model plants such as Arabidopsis thaliana, rice, and Medicago truncatula. Altogether, the INPPO progress and upcoming activities are because of immense support, dedication, and hard work of all members of the INPPO community, and also due to the wide encouragement and support from the communities (scientific and non-scientific).

Proof-of-principle Study to Detect Metabolic Changes in Peritoneal Dialysis Effluent in Patients Who Develop Encapsulating Peritoneal Sclerosis

BACKGROUND: Prolonged peritoneal dialysis (PD) therapy can result in the development of encapsulating peritoneal sclerosis (EPS), characterized by extensive sclerosis of the peritoneum with bowel adhesions often causing obstruction.METHODS: As a proof-of-principle study, holistic profiling of endogenous metabolites has been applied in a prospective collection of PD effluent collected in multiple UK renal centres over 6 years in order to investigate metabolic differences in PD effluent between PD therapy patients who later developed clinically defined EPS (n = 11) and controls, who were matched for PD vintage, age and gender (n = 11).RESULTS: 'Fit-for-purpose' analytical methods employing gas chromatography-mass spectrometry (MS), direct injection MS and quality control samples were developed and validated. These methods were applied in a proof-of-principle study to define metabolic differences in PD effluent related to subsequent development of EPS. Changes in amino acids, amines and derivatives, short-chain fatty acids and derivatives and sugars were observed prior to EPS developing, and changes in the metabolomic profiles could be detected.CONCLUSION: There is potential for applying metabolic profiles to identify patients at risk of developing EPS although long-term prospective studies with larger patient cohorts are required.

Novel Morpholine Ketone Analogs As Potent Histamine H3 Receptor Inverse Agonists with Wake Activity

Structure-activity relationship on a novel ketone class of H(3)R antagonists/inverse agonists is disclosed. Compound 4 showed excellent target potency, selectivity and brain penetration. Evaluation of antagonist 4 in the rat EEG/EMG model demonstrated robust wake activity thereby establishing preclinical proof of concept.

Myocardial Mononuclear Cell Infiltrates Are Not Associated with Increased Serum Cardiac Troponin I in Cynomolgus Monkeys

Myocardial mononuclear cell infiltrate is a spontaneous cardiac finding commonly identified in laboratory cynomolgus monkeys. The infiltrates are predominantly composed of macrophages with lesser lymphocytes and are not typically associated with histologically detectable cardiomyocyte degeneration. These infiltrates are of concern because they confound interpretation of test article-related histopathology findings in nonclinical safety toxicology studies. The interpretation of safety studies would be simplified by a biomarker that could identify myocardial infiltrates prior to animal placement on study. We hypothesized that monkeys with myocardial mononuclear cell infiltrates could be identified before necropsy using an ultrasensitive immunoassay for cardiac troponin I (cTnI). Serum cTnI concentrations in monkeys with myocardial infiltrates were not higher than those in monkeys without infiltrates at any of the sampling times before and on the day of necropsy. Increased serum cTnI levels are not suitable for screening monkeys with myocardial mononuclear cell infiltrates before placement in the study.

Transient Depletion of B Cells in Young Mice Results in Activation of Regulatory T Cells That Inhibit Development of Autoimmune Disease in Adults

B-cell depletion therapy can be effective for treating B-cell lymphomas as well as many human and murine autoimmune diseases. B-cell-deficient mice are normally resistant to spontaneous autoimmune thyroiditis (SAT), but they develop SAT if regulatory T cells are transiently depleted during the first 3-6 weeks after birth. This was also a critical time when B-cell depletion effectively inhibited development of SAT in adult mice. The current study was undertaken to test the hypothesis that transient depletion of B cells using anti-CD20 would be sufficient to suppress SAT if B cells were depleted early in life and that inhibition of SAT would be due to the activity of Treg that functioned most effectively when B cells were absent or low. The results presented here support this hypothesis and indicate that development of autoimmune disease in adults is effectively inhibited when anti-CD20 is administered 1-3 weeks after birth. After 3 weeks, transient B-cell depletion is no longer effective, and B-cell depletion must be maintained to effectively suppress autoimmune disease. B-cell depletion in 1- to 3-week-old mice depletes all B-cell subsets, whereas B-cell depletion initiated in adults spares many marginal zone B cells. Following early B-cell depletion, splenic Treg increase in number, and depletion of Treg reverses the inhibitory effect of anti-CD20 on disease development. Early transient depletion of B cells could be useful for preventing autoimmune disease in individuals at high risk for developing autoimmune diseases as adults.

Industry Influenced Evidence Production in Collaborative Research Communities: A Network Analysis

To measure the relative influence that industry authors have on collaborative research communities and evidence production.

Molecular Profiling of Indolent Human Prostate Cancer: Tackling Technical Challenges to Achieve High-fidelity Genome-wide Data

The contemporary problem of prostate cancer overtreatment can be partially attributed to the diagnosis of potentially indolent prostate cancers that pose low risk to aged men, and lack of sufficiently accurate risk stratification methods to reliably seek out men with indolent diseases. Since progressive acquisition and accumulation of genomic alterations, both genetic and epigenetic, is a defining feature of all human cancers at different stages of disease progression, it is hypothesized that RNA and DNA alterations characteristic of indolent prostate tumors may be different from those previously characterized in the setting of clinically significant prostate cancer. Approaches capable of detecting such alterations on a genome-wide level are the most promising. Such analysis may uncover molecular events defining early initiating stages along the natural history of prostate cancer progression, and ultimately lead to rational development of risk stratification methods for identification of men who can safely forego treatment. However, defining and characterizing indolent prostate cancer in a clinically relevant context remains a challenge, particularly when genome-wide approaches are employed to profile formalin-fixed paraffin-embedded (FFPE) tissue specimens. Here, we provide the conceptual basis underlying the importance of understanding indolent prostate cancer from molecular profiling studies, identify the key hurdles in sample acquisition and variables that affect molecular data derived from FFPE tissues, and highlight recent progresses in efforts to address these technical challenges.Asian Journal of Andrology advance online publication, 6 February 2012; doi:10.1038/aja.2011.147.

Socio-economic Inequalities in Healthy Child Development: the Evidence Grows

Small Interfering RNA Library Screen Identified Polo-like Kinase-1 (PLK1) As a Potential Therapeutic Target for Breast Cancer That Uniquely Eliminates Tumour-initiating Cells

ABSTRACT: INTRODUCTION: Triple negative breast cancers' (TNBCs) high rate of relapse is thought to be due to the presence of tumor-initiating cells (TICs), molecularly defined as being CD44high/CD24-/low. TICs are resilient to chemotherapy and radiation. However, there is no currently accepted molecular target against TNBC and moreover TICs. Therefore, we sought the identification of kinase targets that inhibit TNBC growth and eliminate TICs. METHODS: A genome-wide human kinase siRNA library (691 kinases) was screened against the TNBC cell line SUM149 for growth inhibition. Selected siRNAs were then tested on four different breast cancer cell lines to confirm the spectrum of activity. Their effect on CD44high subpopulation and sorted CD44high/CD24-/low cells of SUM149 was also studied. Further studies were focused on polo-like kinase 1 (PLK1), including its expression in breast cancer cell lines, effect on CD44high/CD24-/low TIC subpopulation, growth inhibition, mammosphere formation and apoptosis as well as the activity of the PLK1 inhibitor, BI 2536. RESULTS: 85 kinases were identified in the screen. 28 of them were further silenced by siRNAs on MDA-MB-231 (TNBC), BT474-M1 (ER+/HER2+, a metastatic variant) and HR5 (ER+/HER2+, a trastuzumab-resistant model) cells and showed broad spectrum of growth inhibition. Importantly, 12/28 kinases also reduced CD44high subpopulation compared to control in SUM149. Further tests of these 12 kinases directly on sorted CD44high/CD24-/low TIC subpopulation of SUM149 cells confirmed their effect. Blocking PLK1 had the greatest growth inhibition on breast cancer cells and TICs by ~80-90% after 72 hours. PLK1 was universally expressed in breast cancer cell lines representing all of the breast cancer subtypes, and was positively correlated to CD44. The PLK1 inhibitor BI 2536 showed similar effects on growth, mammosphere formation and apoptosis as PLK1 siRNAs. Finally, while paclitaxel, doxorubicin and 5-fluorouracil enriched CD44high/CD24-/low population compared to control in SUM149, subsequent treatment with BI 2536 killed the emergent population suggesting it could potentially be used to prevent relapse. CONCLUSION: Inhibiting PLK1 with siRNA or BI 2536 blocked growth of TNBCs including the CD44high/CD24-/low TIC subpopulation and mammosphere formation. Thus, PLK1 could be a potential therapeutic target for the treatment of TNBC as well as other subtypes of breast cancer.

Geriatric Trauma Service: a One-year Experience

Trauma centers nationwide have been experiencing an increase in their elderly trauma patients because of an ever growing elderly population within the United States. Many studies have demonstrated the physiologic differences between an older trauma patient versus a younger trauma patient. Coupling these differences with their coexisting medical comorbidities, makes caring for this population extremely challenging. To meet these challenges, we organized a geriatric trauma unit specifically designed with a multidisciplinary approach to take a more aggressive stance to the care of the geriatric trauma patient.

Branchiobdellidan Infestation on Endangered White-clawed Crayfish (Austropotamobius Pallipes) in the UK

SUMMARYBranchiobdellidans or crayfish worms are clitellate annelids and ectosymbionts of freshwater crayfish. An investigation of branchiobdellidan infestation was undertaken in a population of endangered white-clawed crayfish (Austropotamobius pallipes) in the river Aire, UK. Thirty two percent of animals were infested either by the adult parasite or their cocoons (n=107). Parasite burden increased with host size, but did not differ with sex. Observations of crayfish gill tissue revealed a strong positive relationship between melanization of filaments and parasite prevalence and burden. Taxonomic identification revealed that 1 species of branchiobdellidan was present, Branchiobdella astaci. The first sequences were generated for this species and phylogenetically analysed alongside published sequences for 5 other branchiobdellidan species in Europe. The position of B. astaci within the genus Branchiobdella was confirmed, and it was found to cluster as a sister group to B. parasita.

Allosteric Regulation of Substrate Channeling and Catalysis in the Tryptophan Synthase Bienzyme Complex

The tryptophan synthase α2β2 bi-enzyme complex catalyzes the last two steps in the synthesis of l-tryptophan (l-Trp). The α-subunit catalyzes cleavage of 3-indole-d-glycerol 3'-phosphate (IGP) to give indole and d-glyceraldehyde 3'-phosphate (G3P). Indole is then transferred (channeled) via an interconnecting 25Å-long tunnel, from the α-subunit to the β-subunit where it reacts with l-Ser in a pyridoxal 5'-phosphate-dependent reaction to give l-Trp and a water molecule. The efficient utilization of IGP and l-Ser by tryptophan synthase to synthesize l-Trp utilizes a system of allosteric interactions that (1) function to switch the α-site on and off at different stages of the β-subunit catalytic cycle, and (2) prevent the escape of the channeled intermediate, indole, from the confines of the α- and β-catalytic sites and the interconnecting tunnel. This review discusses in detail the chemical origins of the allosteric interactions responsible both for switching the α-site on and off, and for triggering the conformational changes between open and closed states which prevent the escape of indole from the bienzyme complex.

Prevention of VTE in Nonsurgical Patients: Antithrombotic Therapy and Prevention of Thrombosis, 9th Ed: American College of Chest Physicians Evidence-Based Clinical Practice Guidelines

This guideline addressed VTE prevention in hospitalized medical patients, outpatients with cancer, the chronically immobilized, long-distance travelers, and those with asymptomatic thrombophilia.

Perioperative Management of Antithrombotic Therapy: Antithrombotic Therapy and Prevention of Thrombosis, 9th Ed: American College of Chest Physicians Evidence-Based Clinical Practice Guidelines

This guideline addresses the management of patients who are receiving anticoagulant or antiplatelet therapy and require an elective surgery or procedure.

Carotid Intima-medial Thickness in National Football League Players As an Index of Cardiovascular Disease Risk

Single Molecule Probes of Membrane Structure: Orientation of BODIPY Probes in DPPC As a Function of Probe Structure

Single molecule fluorescence measurements have recently been used to probe the orientation of fluorescent lipid analogs doped into lipid films at trace levels. Using defocused polarized total internal reflection fluorescence microscopy (PTIRF-M), these studies have shown that fluorophore orientation responds to changes in membrane surface pressure and composition, providing a molecular level marker of membrane structure. Here we extend those studies by characterizing the single molecule orientations of six related BODIPY probes doped into monolayers of DPPC. Langmuir-Blodgett monolayers transferred at various surface pressures are used to compare the response from fluorescent lipid analogs in which the location of the BODIPY probe is varied along the length of the acyl chain. For each BODIPY probe location along the chain, comparisons are made between analogs containing phosphocholine and smaller fatty acid headgroups. Together these studies show a general propensity of the BODIPY analogs to insert into membranes with the BODIPY probe aligned along the acyl chains or looped back to interact with the headgroups. For all BODIPY probes studied, a bimodal orientation distribution is observed which is sensitive to surface pressure, with the population of BODIPY probes aligned along the acyl chains increasing with elevated surface pressure. Trends in the single molecule orientations for the six analogs reveal a configuration where optimal placement of the BODIPY probe within the acyl chain maximizes its sensitivity to the surrounding membrane structure. These results are discussed in terms of balancing the effects of headgroup association with acyl chain length in designing the optimal placement of the BODIPY probe.

Undetected Cognitive Impairment and Decision-making Capacity in Patients Receiving Hospice Care

: Cognitive dysfunction is common in patients with advanced, life-threatening illness and can be attributed to a variety of factors (e.g., advanced age, opiate medication). Such dysfunction likely affects decisional capacity, which is a crucial consideration as the end-of-life approaches and patients face multiple choices regarding treatment, family, and estate planning. This study examined the prevalence of cognitive impairment and its impact on decision-making abilities among hospice patients with neither a chart diagnosis of a cognitive disorder nor clinically apparent cognitive impairment (e.g., delirium, unresponsiveness).

Video Incident Analysis of Head Injuries in High School Girls' Lacrosse

Knowledge of injury mechanisms and game situations associated with head injuries in girls' high school lacrosse is necessary to target prevention efforts.

Constipation After Total Hip Replacement May Be Lower with Oral Opioid Analgesia Than with Intravenous Opioid Analgesia

Human Papillomavirus Load Measured by Linear Array Correlates with Quantitative PCR in Cervical Cytology Specimens

Carcinogenic human papillomavirus (HPV) infections are necessary causes of most anogenital cancers. Viral load has been proposed as a marker for progression to cancer precursors but has been confirmed only for HPV16. Challenges in studying viral load are related to the lack of validated assays for a large number of genotypes. We compared viral load measured by Linear Array (LA) HPV genotyping with the gold standard, quantitative PCR (Q-PCR). LA genotyping and Q-PCR were performed in 143 cytology specimens from women referred to colposcopy. LA signal strength was measured by densitometry. Correlation coefficients and receiver operating characteristic (ROC) analyses were used to evaluate analytical and clinical performance. We observed a moderate to strong correlation between the two quantitative viral load measurements, ranging from an R value of 0.61 for HPV31 to an R value of 0.86 for HPV52. We also observed agreement between visual LA signal strength evaluation and Q-PCR. Both quantifications agreed on the disease stages with highest viral load, which varied by type (cervical intraepithelial neoplasia grade 2 [CIN2] for HPV52, CIN3 for HPV16 and HPV33, and cancer for HPV18 and HPV31). The area under the curve (AUC) for HPV16 Q-PCR at the CIN3 cutoff was 0.72 (P = 0.004), and the AUC for HPV18 LA at the CIN2 cutoff was 0.78 (P = 0.04). Quantification of LA signals correlates with the current gold standard for viral load, Q-PCR. Analyses of viral load need to address multiple infections and type attribution to evaluate whether viral load has clinical value beyond the established HPV16 finding. Our findings support conducting comprehensive studies of viral load and cervical cancer precursors using quantitative LA genotyping data.

HijAkt: The PI3K/Akt Pathway in Virus Replication and Pathogenesis

As obligate parasites of cellular processes, viruses must take over cellular macromolecular machinery. It is also becoming clear that viruses routinely control intracellular signaling pathways through the direct or indirect control of kinases and phosphatases. This control of cellular phosphoproteins is important to promote a variety of viral processes, from control of entry to nuclear function to the stimulation of viral protein synthesis. This review focuses on the takeover of the cellular phosphatidylinositol-3-kinase (PI3K)/Akt signaling pathway by a variety of retroviruses, DNA viruses, and RNA viruses, highlighting the functions ascribed to virus activation of PI3K and Akt activity. This review also describes the role that the PI3K/Akt pathway plays in the host response, noting that it that can trigger anti- as well as proviral functions.

Wake-promoting Agents: Search for Next Generation Modafinil: Part II

In search of a next generation molecule to modafinil, a novel wake promoting agent, we previously disclosed bi-phenyl derived racemate compound (±)-2 as a new generation of wake-promoting agent. Here we describe the profiles of the individual enantiomers (-)-2 and (+)-2, respectively.

Stem Cells in Brain Tumour Development and Therapy- Two-sides of the Same Coin

Primary brain tumours are difficult to manage clinically due to their abilities to invade adjacent tissue and infiltrate distant neuropil. These contribute to challenges in surgical management and also limit the effectiveness of radiotherapy. Despite initial responses to chemotherapy, most tumours become chemo-resistant, leading to relapse. Recent identification and isolation of brain cancer stem cells (BCSCs) have broadened our understanding of the molecular pathogenesis and potential Achilles' heel of brain tumours. BCSCs are thought to drive and propagate the tumour and therefore present an important target for further investigations. This review explores the history of the discovery of BCSCs and the evolving concept of "cancer stem cells" in neuro-oncology. We attempt to present a balanced view on the subject and also to update the readers on the molecular biology of BCSCs. Lastly, we outline the potential strategies to target BCSCs which will translate into specific and effective therapies for brain tumours.

Protective Effect of an Elastase Inhibitor in a Neuromyelitis Optica-like Disease Driven by a Peptide of Myelin Oligodendroglial Glycoprotein

Background: The pathology of neuromyelitis optica (NMO), in contrast to multiple sclerosis, comprises granulocyte infiltrates along extensive lengths of spinal cord, as well as optic nerve. Furthermore, IFN-β treatment worsens NMO. We recently found that experimental autoimmune encephalomyelitis (EAE) induced with Th17 cells is exacerbated by IFN-β, in contrast to disease induced with Th1 where treatment attenuated symptoms. Objective: This study demonstrates the similarities between NMO and Th17 EAE and how neutrophils mediate pathology in Th17 disease. Methods: Levels of blood biomarkers in NMO were assessed by Luminex and ELISA. Effects of IFN-β on neutrophils were assessed by culture assays and immunofluorescence. EAE was induced by transfer of myelin-specific Th1 or Th17 cells and treated with Sivelestat sodium hydrate, a neutrophil elastase inhibitor. Results: We show Th17 cytokines, granulocyte chemokines, type 1 interferon and neutrophil elastase are elevated in patients with definitive NMO. In culture, we find that IFN-β stimulates neutrophils to release neutrophil elastase. In Th17 EAE, we demonstrate neutrophilic infiltration in the optic nerve and spinal cord which was not present in Th1 EAE. Blockade of neutrophil elastase with Sivelestat had efficacy in Th17 EAE but not Th1 EAE. Conclusions: The similarities between Th17 EAE and NMO indicate that this model represents several aspects of NMO. Neutrophils are critical in the pathologies of both Th17-EAE and NMO, and therefore blockade of neutrophil elastase is a promising target in treating NMO.

Exposure to Trimethoprim-sulfamethoxazole is Associated with Hospitalisation for Hyperkalaemia in Older People Treated with Spironolactone

Contrasting Roles for CD4 Vs. CD8 T-cells in a Murine Model of Virally Induced "T1 Black Hole" Formation

MRI is sensitive to tissue pathology in multiple sclerosis (MS); however, most lesional MRI findings have limited correlation with disability. Chronic T1 hypointense lesions or "T1 black holes" (T1BH), observed in a subset of MS patients and thought to represent axonal damage, show moderate to strong correlation with disability. The pathogenesis of T1BH remains unclear. We previously reported the first and as of yet only model of T1BH formation in the Theiler's murine encephalitis virus induced model of acute CNS neuroinflammation induced injury, where CD8 T-cells are critical mediators of axonal damage and related T1BH formation. The purpose of this study was to further analyze the role of CD8 and CD4 T-cells through adoptive transfer experiments and to determine if the relevant CD8 T-cells are classic epitope specific lymphocytes or different subsets. C57BL/6 mice were used as donors and RAG-1 deficient mice as hosts in our adoptive transfer experiments. In vivo 3-dimensional MRI images were acquired using a 7 Tesla small animal MRI system. For image analysis, we used semi-automated methods in Analyze 9.1; transfer efficiency was monitored using FACS of brain infiltrating lymphocytes. Using a peptide depletion method, we demonstrated that the majority of CD8 T-cells are classic epitope specific cytotoxic cells. CD8 T-cell transfer successfully restored the immune system's capability to mediate T1BH formation in animals that lack adaptive immune system, whereas CD4 T-cell transfer results in an attenuated phenotype with significantly less T1BH formation. These findings demonstrate contrasting roles for these cell types, with additional evidence for a direct pathogenic role of CD8 T-cells in our model of T1 black hole formation.

Modeling the Impact of Changing Patient Transportation Systems on Peri-operative Process Performance in a Large Hospital: Insights from a Computer Simulation Study

Transportation of patients is a key hospital operational activity. During a large construction project, our patient admission and prep area will relocate from immediately adjacent to the operating room suite to another floor of a different building. Transportation will require extra distance and elevator trips to deliver patients and recycle transporters (specifically: personnel who transport patients). Management intuition suggested that starting all 52 first cases simultaneously would require many of the 18 available elevators. To test this, we developed a data-driven simulation tool to allow decision makers to simultaneously address planning and evaluation questions about patient transportation. We coded a stochastic simulation tool for a generalized model treating all factors contributing to the process as JAVA objects. The model includes elevator steps, explicitly accounting for transporter speed and distance to be covered. We used the model for sensitivity analyses of the number of dedicated elevators, dedicated transporters, transporter speed and the planned process start time on lateness of OR starts and the number of cases with serious delays (i.e., more than 15 min). Allocating two of the 18 elevators and 7 transporters reduced lateness and the number of cases with serious delays. Additional elevators and/or transporters yielded little additional benefit. If the admission process produced ready-for-transport patients 20 min earlier, almost all delays would be eliminated. Modeling results contradicted clinical managers' intuition that starting all first cases on time requires many dedicated elevators. This is explained by the principle of decreasing marginal returns for increasing capacity when there are other limiting constraints in the system.

Cancer Immunoediting in Malignant Glioma

ABSTRACT: Significant work from many laboratories over the last decade in the study of cancer immunology has resulted in the development of the cancer immunoediting hypothesis. This contemporary framework of the naturally-arising immune system-tumor interaction is thought to comprise 3 phases: elimination, wherein immunity subserves an extrinsic tumor suppressor function and destroys nascent tumor cells; equilibrium, wherein tumor cells are constrained in a period of latency under immune control; and escape, wherein tumor cells outpace immunity and progress clinically. In this review, we will address in detail the relevance of the cancer immunoediting concept to neurosurgeons and neuro-oncologists treating and studying malignant glioma by exploring the de novo immune response to these tumors, how these tumors may persist in vivo, the mechanisms by which these cells may escape/attenuate immunity, and, ultimately, how this concept may influence our immunotherapeutic approaches.

Breathe Easier Online: Evaluation of a Randomized Controlled Pilot Trial of an Internet-based Intervention to Improve Well-being in Children and Adolescents with a Chronic Respiratory Condition

Chronic respiratory illnesses are the most common group of childhood chronic health conditions and are overrepresented in socially isolated groups.

Spontaneous Olfactory Receptor Neuron Activity Determines Follower Cell Response Properties

Noisy or spontaneous activity is common in neural systems and poses a challenge to detecting and discriminating signals. Here we use the locust to answer fundamental questions about noise in the olfactory system: Where does spontaneous activity originate? How is this activity propagated or reduced throughout multiple stages of neural processing? What mechanisms favor the detection of signals despite the presence of spontaneous activity? We found that spontaneous activity long observed in the secondary projection neurons (PNs) originates almost entirely from the primary olfactory receptor neurons (ORNs) rather than from spontaneous circuit interactions in the antennal lobe, and that spontaneous activity in ORNs tonically depolarizes the resting membrane potentials of their target PNs and local neurons (LNs) and indirectly tonically depolarizes tertiary Kenyon cells (KCs). However, because these neurons have different response thresholds, in the absence of odor stimulation, ORNs and PNs display a high spontaneous firing rate but KCs are nearly silent. Finally, we used a simulation of the olfactory network to show that discrimination of signal and noise in the KCs is best when threshold levels are set so that baseline activity in PNs persists. Our results show how the olfactory system benefits from making a signal detection decision after a point of maximal information convergence, e.g., after KCs pool inputs from many PNs.

Geriatric Inflammatory Bowel Disease: Phenotypic Presentation, Treatment Patterns, Nutritional Status, Outcomes, and Comorbidity

BACKGROUND AND AIMS: The U.S. population is aging and the burden of geriatric inflammatory bowel disease (IBD) patients has increased. Systematic data describing phenotypic presentation, treatment regimens, outcomes and comorbidities in elderly IBD patients is limited. We performed a retrospective observational study of IBD patients age ≥65 followed in a 20-hospital system to determine patterns of phenotypic presentation, treatment, polypharmacy, nutritional status and comorbidity. METHODS: Data were extracted from electronic medical record based on ICD-9 coding/indexed terms on Crohn's disease (CD) and ulcerative colitis (UC) patients. RESULTS: A total of 393 geriatric IBD patients were identified (49.1% males; 50.9% females; 61.8% UC; 38.2% CD; 73.4 ± 6.6 years old). Younger age at diagnosis of CD (≤64) was associated with greater prevalence of small bowel surgeries (63.6%) compared with those diagnosed after age ≥65 (20.9%) (p < 0.005). Fistulizing/penetrating disease was frequent in patients diagnosed with CD at a younger age (43.6% compared to 7%) (p < 0.005). IBD maintenance treatment included: 44% 5-ASA agents; 31.6% maintenance prednisone (defined as ≥6 months treatment duration); 4.8% steroid suppositories; 5.6% 6MP/azathioprine; 1.3% methotrexate; 1.3% adalimumab; 1.3% infliximab; 9.4% loperamide/diphenoxylate/atropine; 0.5% had no IBD medications. Longer duration of CD disease correlated with vitamin B12, vitamin D and iron deficiency. CONCLUSION: Geriatric patients diagnosed with CD earlier in life had greater small bowel involvement compared with new onset geriatric CD. There is low utilization of immunomodulator and biologic agents in geriatric IBD patients. Duration of CD correlates with nutrient deficiency. Prospective studies are warranted in this respect.

Predatory Functional Response and Prey Choice Identify Predation Differences Between Native/invasive and Parasitised/unparasitised Crayfish

Invasive predators may change the structure of invaded communities through predation and competition with native species. In Europe, the invasive signal crayfish Pacifastacus leniusculus is excluding the native white clawed crayfish Austropotamobius pallipes.

Obesity Inequality in Malaysia: Decomposing Differences by Gender and Ethnicity Using Quantile Regression

Objective. Obesity prevalence is unequally distributed across gender and ethnic group in Malaysia. In this paper, we examine the role of socioeconomic inequality in explaining these disparities. Design. The body mass index (BMI) distributions of Malays and Chinese, the two largest ethnic groups in Malaysia, are estimated through the use of quantile regression. The differences in the BMI distributions are then decomposed into two parts: attributable to differences in socioeconomic endowments and attributable to differences in responses to endowments. Results. For both males and females, the BMI distribution of Malays is shifted toward the right of the distribution of Chinese, i.e., Malays exhibit higher obesity rates. In the lower 75% of the distribution, differences in socioeconomic endowments explain none of this difference. At the 90th percentile, differences in socioeconomic endowments account for no more than 30% of the difference in BMI between ethnic groups. Conclusions. Our results demonstrate that the higher levels of income and education that accrue with economic development will likely not eliminate obesity inequality. This leads us to conclude that reduction of obesity inequality, as well the overall level of obesity, requires increased efforts to alter the lifestyle behaviors of Malaysians.

Development of Metronidazole-resistant Lines of Blastocystis Sp

Metronidazole (MTR) is frequently used for the treatment of Blastocystis infections, but with variable effectiveness, and often with treatment failures as a possible result of drug resistance. We have developed two Blastocystis MTR-resistant (MTR(R)) subtype 4 WR1 lines (WR1-M4 and WR1-M5), with variable susceptibility to a panel of anti-protozoal agents including various 5-nitroimidazoles, nitazoxanide and furazolidone. WR1-M4 and WR1-M5 were developed and assessed over an 18-month period and displayed persistent MTR resistance, being more than 2.5-fold less susceptible to MTR than the parent isolate. The MTR(R) lines grew with a similar g time to WR1, but were morphologically less consistent with a mixture of size. All Blastocystis isolates and the MTR(R) lines were most susceptible to the 5-nitroimidazole drug ronidazole. WR1-M5 was apparently cross-resistant to satranidazole and furazolidone, and WR1-M4 was cross-resistant to nitazoxanide. These MTR(R) lines now provide a valuable tool for the continued assessment of the efficacy and mechanism of action of new and established drugs against a range of Blastocystis sp. subtypes, in order to identify a universally effective drug and to facilitate understanding of the mechanisms of drug action and resistance in Blastocystis.

Feedback-Based Alcohol Interventions for Mandated Students: An Effectiveness Study of Three Modalities

The present study used a randomized clinical trial design to examine the effectiveness of personalized alcohol feedback delivered individually, in a group and via computer on alcohol use and related negative consequences in a sample of 173 college students referred for alcohol-related violations. Findings revealed statistically significant reductions in alcohol use and related harms for the individually delivered intervention, with significant reductions in alcohol-related harms for the electronically delivered intervention. No statistically significant results were found for the group-delivered intervention or between groups, and a main effect of time was noted for all outcome variables. This study adds to the literature by being the first randomized clinical trial to include analyses of an empirically supported individually delivered personalized alcohol feedback intervention with more cost-effective group-delivered and electronically delivered feedback formats within a single research design, by expanding the range of participant drinking habits reported at baseline to include all drinking levels and not solely those classified as 'heavy drinking' and by providing anonymity pre-intervention and post-intervention given the potential demand characteristics to underreport illegal and/or illicit behaviours in this vulnerable population. Copyright © 2012 John Wiley & Sons, Ltd. KEY PRACTITIONER MESSAGE: Personalized alcohol feedback delivered in a one-on-one, face-to-face format serves to decrease both alcohol use and harms in mandated college students. The use of web-delivered personalized alcohol feedback may be clinically useful when working with a mandated student population to reduce alcohol-related harms. Personalized alcohol feedback delivered in a group setting may not be indicated for use with a mandated student population as it does not demonstrate decreases in either alcohol use or harms, possibly because of the normalization of deviant behaviour.

Current Approaches to Journal Club by General Surgery Programs Within the Southwestern Surgical Congress

Journal club (JC) is a well-recognized education tool for many postgraduate medical education programs. Journal club helps residents learn critical analytic skills and keep up to date with current medical practices. To our knowledge, there is minimal evidence in the current literature detailing modern JC practices of general surgery training programs. Our study attempts to define how general surgery residency programs are implementing JC in their training process.

A Novel Combination of Printed 3-Dimensional Anatomic Templates and Computer-assisted Surgical Simulation for Virtual Preoperative Planning in Charcot Foot Reconstruction

Charcot foot syndrome (Charcot neuroarthropathy affecting the foot), particularly in its latter stages, may pose a significant technical challenge to the surgeon. Because of the lack of anatomic consistency, preoperative planning with virtual and physical models of the foot could improve the chances of achieving a predictable intraoperative result. In this report, we describe the use of a novel, inexpensive, 3-dimensional template printing technique that can provide, with just a normal printer, multiple "copies" of the foot to be repaired. Although we depict this method as it pertains to repair of the Charcot foot, it could also be used to plan and practice, or revise, 3-dimensional surgical manipulations of other complex foot deformities.

The Expectancy Challenge Alcohol Literacy Curriculum (ECALC): A Single Session Group Intervention to Reduce Alcohol Use

The Expectancy Challenge Alcohol Literacy Curriculum (ECALC) is a single session group-delivered program designed to modify alcohol expectancy processes and reduce alcohol use among children and young adults. The objective of this study was to demonstrate the effectiveness of the ECALC in reducing risky alcohol use among heavy drinking college men. Four fraternities at a large state university were randomly assigned to receive either the single session ECALC or a control presentation (2 fraternity houses per condition, n = 250). Alcohol expectancies were assessed before and immediately after program presentation. Results demonstrated significant changes on 5 of the 7 subscales of the Comprehensive Effects of Alcohol Scale (CEOA) among students who received the ECALC when compared with control participants. Alcohol use data were collected for 4 weeks before and 4 weeks after program presentation. Compared with those in the control condition, students who received the ECALC demonstrated significant reductions in all facets of alcohol use measured, including decreased mean and peak blood alcohol content (BAC), decreased mean number of days drinking per week, decreased mean drinks per sitting, and decreased number of binge-drinking episodes per month. This study represents 2 important advances. First is the significant reduction in risky alcohol use produced by a single session group-delivered program. The second important advance is the success in changing expectancy processes without using impractical elements common in previous expectancy challenge methods (e.g., a "barlab" environment and actual alcohol administration). (PsycINFO Database Record (c) 2012 APA, all rights reserved).

Analysis of the Saccharomyces Cerevisiae Pan-genome Reveals a Pool of Copy Number Variants Distributed in Diverse Yeast Strains from Differing Industrial Environments

Although the budding yeast Saccharomyces cerevisiae is arguably one of the most well-studied organisms on earth, the genome-wide variation within this species-i.e., its "pan-genome"-has been less explored. We created a multispecies microarray platform containing probes covering the genomes of several Saccharomyces species: S. cerevisiae, including regions not found in the standard laboratory S288c strain, as well as the mitochondrial and 2-μm circle genomes-plus S. paradoxus, S. mikatae, S. kudriavzevii, S. uvarum, S. kluyveri, and S. castellii. We performed array-Comparative Genomic Hybridization (aCGH) on 83 different S. cerevisiae strains collected across a wide range of habitats; of these, 69 were commercial wine strains, while the remaining 14 were from a diverse set of other industrial and natural environments. We observed interspecific hybridization events, introgression events, and pervasive copy number variation (CNV) in all but a few of the strains. These CNVs were distributed throughout the strains such that they did not produce any clear phylogeny, suggesting extensive mating in both industrial and wild strains. To validate our results and to determine whether apparently similar introgressions and CNVs were identical by descent or recurrent, we also performed whole-genome sequencing on nine of these strains. These data may help pinpoint genomic regions involved in adaptation to different industrial milieus, as well as shed light on the course of domestication of S. cerevisiae.

Proteasome Inhibitors for Cancer Therapy

Proteasome, a large multicatalytic proteinase complex that plays an important role in processing of proteins, has been shown to possess multiple catalytic activities. Among its various activities, the 'chymotrypsin-like' activity of proteasome has emerged as the focus of drug discovery efforts in cancer therapy. Herein we report chiral boronate derived novel, potent, selective and cell-permeable peptidomimetic inhibitors 6 and 7 that displayed activity against various rodent and human tumor cell lines (in vitro).

Cancer: Solving an Age-old Problem

Q&A: Controversy and Intellect. Interview by Julie Corliss

The Role and Impact of Research Agendas on the Comparative-effectiveness Research Among Antihyperlipidemics

Although it is well established that funding source influences the publication of clinical trials, relatively little is known about how funding influences trial design. We examined a public trial registry to determine how funding source shapes trial design among trials involving antihyperlipidemics. We used an automated process to identify and analyze 809 trials from a set of 72,564. Three networks representing industry-, collaboratively, and non-industry-funded trials were constructed. Each network comprised 18 drugs as nodes connected according to the number of comparisons made between them. The results indicated that industry-funded trials were more likely to compare across drugs and examine dyslipidemia as a condition, and less likely to register safety outcomes. The source of funding for clinical trials had a measurable effect on trial design, which helps quantify differences in research agendas. Improved monitoring of current clinical trials may be used to more closely align research agendas to clinical needs.

Mindful Pregnancy and Childbirth: Effects of a Mindfulness-based Intervention on Women's Psychological Distress and Well-being in the Perinatal Period

This pilot study explored the effects of an 8-week mindfulness-based cognitive therapy group on pregnant women. Participants reported a decline in measures of depression, stress and anxiety; with these improvements continuing into the postnatal period. Increases in mindfulness and self-compassion scores were also observed over time. Themes identified from interviews describing the experience of participants were: 'stop and think', 'prior experience or expectations', 'embracing the present', 'acceptance' and 'shared experience'. Childbirth preparation classes might benefit from incorporating training in mindfulness.

Quantitative Analysis of Tooth Surface Loss Associated with Gastroesophageal Reflux Disease: a Longitudinal Clinical Study

Acid regurgitation resulting from gastroesophageal reflux disease (GERD) causes dissolution of tooth structure. The authors conducted a longitudinal clinical study to measure tooth surface loss associated with GERD.

Conservation. Reconsidering the Consequences of Selective Fisheries

Strain Variation in the Transcriptome of the Dengue Fever Vector, Aedes Aegypti

Studies of transcriptome dynamics provide a basis for understanding functional elements of the genome and the complexity of gene regulation. The dengue vector mosquito, Aedes aegypti, exhibits great adaptability to diverse ecological conditions, is phenotypically polymorphic, and shows variation in vectorial capacity to arboviruses. Previous genome sequencing showed richness in repetitive DNA and transposable elements that can contribute to genome plasticity. Population genetic studies revealed a varying degree of worldwide genetic polymorphism. However, the extent of functional genetic polymorphism across strains is unknown. The transcriptomes of three Ae. aegypti strains, Chetumal (CTM), Rexville D-Puerto Rico (Rex-D) and Liverpool (LVP), were compared. CTM is more susceptible than Rex- D to infection by dengue virus serotype 2. A total of 4188 transcripts exhibit either no or small variation (<2-fold) among sugar-fed samples of the three strains and between sugar- and blood-fed samples within each strain, corresponding most likely to genes encoding products necessary for vital functions. Transcripts enriched in blood-fed mosquitoes encode proteins associated with catalytic activities, molecular transport, metabolism of lipids, carbohydrates and amino acids, and functions related to blood digestion and the progression of the gonotropic cycle. Significant qualitative and quantitative differences were found in individual transcripts among strains including differential representation of paralogous gene products. The majority of immunity-associated transcripts decreased in accumulation after a bloodmeal and the results are discussed in relation to the different susceptibility of CTM and Rex-D mosquitoes to DENV2 infection.

Ipsilateral Diaphyseal Femur Fracture After Resection Arthroplasty

Resection arthroplasty of the hip is most commonly used for recalcitrant infections in the setting of prior hemiarthroplasty and total hip arthroplasty. Reported complications of this procedure include shortening of the extremity and ambulation difficulties requiring assistive devices. We report a case of an unusual finding of an ipsilateral femoral shaft fracture after a ground level fall 4 months after a resection arthroplasty for a septic hip. A closed reduction with internal fixation using an intramedullary nail was performed, and 3 months postoperatively, the patient was ambulating with device assistance.

CMV Infection Attenuates the Disease Course in a Murine Model of Multiple Sclerosis

Recent evidence in multiple sclerosis (MS) suggests that active CMV infection may result in more benign clinical disease. The goal of this pilot study was to determine whether underlying murine CMV (MCMV) infection affects the course of the Theiler's murine encephalitis virus (TMEV) induced murine model of MS. A group of eight TMEV-infected mice were co-infected with MCMV at 2 weeks prior to TMEV infection while a second group of TMEV-infected mice received MCMV two weeks post TMEV. We also used 2 control groups, where at the above time points MCMV was replaced with PBS. Outcome measures included (1) monthly monitoring of disability via rotarod for 8 months; (2) in vivo MRI for brain atrophy studies and (3) FACS analysis of brain infiltrating lymphocytes at 8 months post TMEV infection. Co-infection with MCMV influenced the disease course in mice infected prior to TMEV infection. In this group, rotarod detectable motor performance was significantly improved starting 3 months post-infection and beyond (p≤0.024). In addition, their brain atrophy was close to 30% reduced at 8 months, but this was only present as a trend due to low power (p = 0.19). A significant reduction in the proportion of brain infiltrating CD3+ cells was detected in this group (p = 0.026), while the proportion of CD45+ Mac1+ cells significantly increased (p = 0.003). There was also a strong trend for a reduced proportion of CD4+ cells (p = 0.17) while CD8 and B220+ cell proportion did not change. These findings support an immunomodulatory effect of MCMV infection in this MS model. Future studies in this co-infection model will provide insight into mechanisms which modulate the development of demyelination and may be utilized for the development of novel therapeutic strategies.

Robot-assisted Transhiatal Esophagectomy: a 3-year Single-center Experience

Minimally invasive esophagectomy has emerged as an important procedure for disease management in esophageal cancer (EC) with clear margin status, less morbidity, and shorter hospital stays compared with open procedures. The experience with transhiatal approach robotic esophagectomy (RE) for dissection of thoracic esophagus and associated morbidity is described here. Between March 2007 and November 2010, 40 patients with resectable esophageal indications underwent transhiatal RE at the institute. Clinical data for all patients were collected prospectively. Of 40 patients undergoing RE, one patient had an extensive benign stricture, one had high-grade dysplasia, and 38 had EC. Five patients were converted from robotic to open. Median operative time and estimated blood loss were 311 minutes and 97.2 mL, respectively. Median intensive care unit stay was 1 day (range, 0-16), and median length of hospital stay was 9 days (range, 6-36). Postoperative complications frequently observed were anastomotic stricture (n= 27), recurrent laryngeal nerve paresis (n= 14), anastomotic leak (n= 10), pneumonia (n= 8), and pleural effusion (n= 18). Incidence rates of laryngeal nerve paresis (35%) and leak rate (25%) were somewhat higher in comparison with that reported in literature. However, all vocal cord injuries were temporary, and all leaks healed following opening of the cervical incision and drainage. None of the patients died in the hospital, and 30-day mortality was 2.5% (1/40). Median number of lymph nodes removed was 20 (range, 3-38). In 33 patients with known lymph node locations, median of four (range, 0-12) nodes was obtained from the mediastinum, and median of 15 (range, 1-26) was obtained from the abdomen. R0 resection was achieved in 94.7% of patients. At the end of the follow-up period, 25 patients were alive, 13 were deceased, and 2 patients were lost to follow-up. For patients with EC, median disease-free survival was 20 months (range, 3-45). Transhiatal RE, by experience, is a feasible albeit evolving oncologic operation with low hospital mortality. The benefits include minimally invasive mediastinal dissection without thoracotomy or thoracoscopy. A reasonable operative time with minimal blood loss and postoperative morbidity can be achieved, in spite of the technically demanding nature of the procedure. Broader use of this technology in a setting of high-volume comprehensive surgical programs will almost certainly reduce the complication rates. Robotic tanshiatal esophagectomy with the elimination of a thoracic approach should be considered an option for the appropriate patient population in a comprehensive esophageal program.

Total Pancreatectomy and Islet Autotransplantation for Chronic Pancreatitis

Total pancreatectomy (TP) with intraportal islet autotransplantation (IAT) can relieve pain and preserve β-cell mass in patients with chronic pancreatitis (CP) when other therapies fail. We report on a >30-year single-center series.

Recovery of the Hypothalamic-Pituitary-Adrenal Axis in Children and Adolescents After Surgical Cure of Cushing's Disease

Context:Recovery of the hypothalamic-pituitary-adrenal axis (HPAA) after transsphenoidal surgery (TSS) for Cushing's disease (CD) in children has not been adequately studied.Objective:Our objective was to assess time to recovery of the HPAA after TSS in children with CD.Design and Setting:This was a case series at the National Institutes of Health Clinical Center.Patients:Fifty-seven patients with CD (6-18 yr, mean 13.0 ± 3.1 yr) given a standard regimen of glucocorticoid tapering after TSS were studied out of a total of 73 recruited.Interventions:ACTH (250 μg) stimulation tests were administered at approximately 6-month intervals for up to 36 months. Age, sex, pubertal status, body mass index, length of disease, midnight cortisol, and urinary free cortisol at diagnosis were analyzed for effects on recovery.Main Outcome Measure:The main outcome measure was complete recovery of the HPAA as defined by a cortisol level of at least 18 μg/dl in response to 250 μg ACTH.Results:Full recovery was reached by 43 (75.4%) of 57 patients, with 29 of the 43 (67.4%) and 41 of the 43 (95.3%) recovering by 12 and 18 months, respectively. The overall mean time to recovery was 12.6 ± 3.3 months. Kaplan-Meier survivor function estimated a 50% chance of recovering by 12 months after TSS and 75% chance of recovering within 14 months. By receiver operating characteristic curve assessment, the cutoff of at least 10-11 μg/dl of cortisol as the peak of ACTH stimulation testing at 6 months after TSS yielded the highest sensitivity (70-80%) and specificity (64-73%) to predict full recovery of the HPAA at 12 months. Two of the four patients that recovered fully within 6 months had recurrent CD.Conclusions:Although this is not a randomized study, we present our standardized tapering regimen for glucocorticoid replacement after TSS that led to recovery of the HPAA in most patients within the first postoperative year. Multiple factors may affect this process, but an early recovery may indicate disease recurrence.

Complex Traits Analysis of Chicken Growth Using Targeted Genetical Genomics

Dissecting the genetic control of complex trait variation remains very challenging, despite many advances in technology. The aim of this study was to use a major growth quantitative trait locus (QTL) in chickens mapped to chromosome 4 as a model for a targeted approach to dissect the QTL. We applied a variant of the genetical genomics approach to investigate genome-wide gene expression differences between two contrasting genotypes of a marked QTL. This targeted approach allows the direct quantification of the link between the genotypes and the genetic responses, thus narrowing the QTL-phenotype gap using fewer samples (i.e. microarrays) compared with the genome-wide genetical genomics studies. Four differentially expressed genes were localized under the region of the QTL. One of these genes is a potential positional candidate gene (AADAT) that affects lysine and tryptophan metabolism and has alternative splicing variants between the two genotypes. In addition, the lysine and glycolysis metabolism pathways were significantly enriched for differentially expressed genes across the genome. The targeted approach provided a complementary route to fine mapping of QTL by characterizing the local and the global downstream effects of the QTL and thus generating further hypotheses about the action of that QTL.

Surgical Palliative Care: Recent Trends and Developments

Palliation has been an essential, if not the primary, activity of surgery during much of its history. However, it has been only during the past decade that the modern principles and practices of palliative care developed in the nonsurgical specialties in the United States and abroad have been introduced to surgical institutions, widely varied practice settings, education, and research.

Update on Surgical Palliative Care

Accelerated Evolution of Mitochondrial but Not Nuclear Genomes of Hymenoptera: New Evidence from Crabronid Wasps

Mitochondrial genes in animals are especially useful as molecular markers for the reconstruction of phylogenies among closely related taxa, due to the generally high substitution rates. Several insect orders, notably Hymenoptera and Phthiraptera, show exceptionally high rates of mitochondrial molecular evolution, which has been attributed to the parasitic lifestyle of current or ancestral members of these taxa. Parasitism has been hypothesized to entail frequent population bottlenecks that increase rates of molecular evolution by reducing the efficiency of purifying selection. This effect should result in elevated substitution rates of both nuclear and mitochondrial genes, but to date no extensive comparative study has tested this hypothesis in insects. Here we report the mitochondrial genome of a crabronid wasp, the European beewolf (Philanthus triangulum, Hymenoptera, Crabronidae), and we use it to compare evolutionary rates among the four largest holometabolous insect orders (Coleoptera, Diptera, Hymenoptera, Lepidoptera) based on phylogenies reconstructed with whole mitochondrial genomes as well as four single-copy nuclear genes (18S rRNA, arginine kinase, wingless, phosphoenolpyruvate carboxykinase). The mt-genome of P. triangulum is 16,029 bp in size with a mean A+T content of 83.6%, and it encodes the 37 genes typically found in arthropod mt genomes (13 protein-coding, 22 tRNA, and two rRNA genes). Five translocations of tRNA genes were discovered relative to the putative ancestral genome arrangement in insects, and the unusual start codon TTG was predicted for cox2. Phylogenetic analyses revealed significantly longer branches leading to the apocritan Hymenoptera as well as the Orussoidea, to a lesser extent the Cephoidea, and, possibly, the Tenthredinoidea than any of the other holometabolous insect orders for all mitochondrial but none of the four nuclear genes tested. Thus, our results suggest that the ancestral parasitic lifestyle of Apocrita is unlikely to be the major cause for the elevated substitution rates observed in hymenopteran mitochondrial genomes.

Narrow Band Imaging of Squamous Cell Carcinoma Tumors Using Topically Delivered Anti-EGFR Antibody Conjugated Gold Nanorods

Nanoparticles have recently gained interest as exogenous contrast agents in a variety of biomedical applications related to cancer detection and treatment. The objective of this study was to determine the potential of topically administered antibody conjugated gold nanorods (GNRs) for imaging squamous cell carcinomas (SCCs) of the skin using near-infrared narrowband imaging (NBI). Near-infrared (NIR) NBI images narrow wavelength bands to enhance contrast from plasmonic particles in a wide field portable and noncontact device that is clinically compatible for real-time tumor imaging and tumor margin demarcation.

Polo-Like Kinase 1 (PLK1) Inhibition Kills Glioblastoma Multiforme Brain Tumour Cells in Part Through Loss of SOX2 and Delays Tumour Progression in Mice

Glioblastoma multiforme (GBM) ranks amongst the deadliest types of cancer and given this new therapies are urgently needed. To identify molecular targets, we queried a microarray profiling 467 human GBMs and discovered that polo-like kinase 1 (PLK1) was highly expressed in these tumours and that it clustered with the proliferative subtype. Patients with PLK1-high tumours were more likely to die from their disease suggesting that current therapies are inactive against such tumours. This prompted us to examine its expression in brain tumour initiating cells (BTICs) given their association with treatment failure. BTICs isolated from patients expressed 110-470 times more PLK1 than normal human astrocytes. Moreover, BTICs rely on PLK1 for survival because the PLK1 inhibitor BI2536 inhibited their growth in tumoursphere cultures. PLK1 inhibition suppressed growth, caused G(2) /M arrest, induced apoptosis and reduced the expression of SOX2, a marker of neural stem cells, in SF188 cells. Consistent with SOX2 inhibition, the loss of PLK1 activity caused the cells to differentiate based on elevated levels of GFAP and changes in cellular morphology. We then knocked-down SOX2 with siRNA and showed that it too inhibited cell growth and induced cell death. Likewise, in U251 cells, PLK1 inhibition suppressed cell growth, down-regulated SOX2 and induced cell death. Furthermore, BI2536 delayed tumour growth of U251 cells in an orthotopic brain tumour model, demonstrating that the drug is active against GBM. In conclusion, PLK1 level is elevated in GBM and its inhibition restricts the growth of brain cancer cells.

Effect of Misclassification of Antiretroviral Treatment Status on the Prevalence of Transmitted HIV-1 Drug Resistance

ABSTRACT: BACKGROUND: Estimates of the prevalence of transmitted HIV drug resistance (TDR) in a population are derived from resistance tests performed on samples from patients thought to be naive to antiretroviral treatment (ART). Much of the debate over reliability of estimates of the prevalence of TDR has focused on whether the sample population is representative. However estimates of the prevalence of TDR will also be distorted if some ART-experienced patients are misclassified as ART-naive. METHODS: The impact of misclassification bias on the rate of TDR was examined. We developed methods to obtain adjusted estimates of the prevalence of TDR for different misclassification rates, and conducted sensitivity analyses of trends in the prevalence of TDR over time using data from the UK HIV Drug Resistance Database. Logistic regression was used to examine trends in the prevalence of TDR over time. RESULTS: The observed rate of TDR was higher than true TDR when misclassification was present and increased as the proportion of misclassification increased. As the number of naive patients with a resistance test relative to the number of experienced patients with a test increased, the difference between true and observed TDR decreased. The observed prevalence of TDR in the UK reached a peak of 11.3% in 2002 (odds of TDR increased by 1.10 (95% CI 1.02, 1.19, p(linear trend) = 0.02) per year 1997-2002) before decreasing to 7.0% in 2007 (odds of TDR decreased by 0.90 (95% CI 0.87, 0.94, p(linear trend) <0.001) per year 2002-2007. Trends in adjusted TDR were altered as the misclassification rate increased; the significant downward trend between 2002-2007 was lost when the misclassification increased to over 4%. CONCLUSION: The effect of misclassification of ART on estimates of the prevalence of TDR may be appreciable, and depends on the number of naive tests relative to the number of experienced tests. Researchers can examine the effect of ART misclassification on their estimates of the prevalence of TDR if such a bias is suspected.

Myosin-X Functions in Polarized Epithelial Cells

Myosin-X (Myo10) is an unconventional myosin that localizes to the tips of filopodia and has critical functions in filopodia. Although Myo10 has been studied primarily in non-polarized, fibroblast-like cells, Myo10 is expressed in vivo in many epithelia-rich tissues such as kidney. Here, we investigate the localization and functions of Myo10 in polarized epithelial cells, using MDCK (Madin-Darby Canine Kidney II) cells as a model system. Calcium-switch experiments demonstrate that, during junction assembly, GFP-Myo10 localizes to lateral membrane cell-cell contacts and to filopodia-like structures imaged by TIRF on the basal surface. Knockdown of Myo10 leads to delayed recruitment of E-cadherin and ZO-1 to junctions, as well as a delay in tight junction barrier formation as indicated by a delay in the development of peak transepithelial electrical resistance (TER). Although Myo10 knockdown cells eventually mature into monolayers with normal TER, these monolayers do exhibit increased paracellular permeability to fluorescent dextrans. Importantly, knockdown of Myo10 leads to mitotic spindle misorientation, and in 3D culture, Myo10 knockdown cysts exhibit defects in lumen formation. Together, these results reveal that Myo10 functions in polarized epithelial cells in junction formation, regulation of paracellular permeability, and epithelial morphogenesis.

It's Not My Fault: Understanding Nursing Students' Causal Attributions in Pathophysiology

Pathophysiology is a difficult subject matter for many nursing students. This course is also critical for safe clinical practice. However, little research has explored what variables may influence nursing students' success in this course. This study is the first in a forthcoming series that seeks to better understand how to facilitate student success in Pathophysiology. In this study, students' causal attributions for successes and failures were explored as these attributions greatly influence future academic motivation and behavior. Students were asked to respond to two open-ended questions in order to better understand what causal attributions students were making for their successes and failures in Pathophysiology. Seventy-five Bachelor of Science in Nursing students who were enrolled in Pathophysiology returned their responses (92.6% response rate). Content analysis was utilized to determine whether students were making internal or external causal attributions for their successes and failures. Additionally, responses were evaluated in order to identify common themes shared by respondents. The majority of respondents (84%) attributed their academic successes in Pathophysiology in part to internal causes, and the majority of respondents (68%) attributed their academic failures, in part, to external causes. In this study the majority of students attributed their successes to controllable, unstable causes-primarily effort. Research indicates that attributing success to effort may reflect that students' confidence in their abilities is suffering, and that attributing failures to external causes, such as task difficulty, are also detrimental to performance and learning (Siegle et al., 2009). The results of this study are further presented and discussed.

"Thinking About It for Somebody Else": Alzheimer's Disease Research and Proxy Decision Makers' Translation of Ethical Principles Into Practice

OBJECTIVES:: Ethical guidelines suggest that, when enrolling patients with dementia in research, alterative decision makers (proxies) should base their decision on a "substituted judgment" of how the patient would have decided. If unable to make a substituted judgment, proxies are asked to decide on the basis of the patient's best interests. This mixed-methods study is the first to examine explicitly whether and to what degree proxies differentiate between these two approaches and what considerations influence their mode of decision making. DESIGN:: Interview study regarding enrollment of relative in hypothetical clinical trial of an investigational drug for Alzheimer disease. Participants were randomized to respond to questions about one of four hypothetical clinical trials that differed by levels of described risk and potential benefit. PARTICIPANTS:: Proxy decision makers (N = 40). MEASUREMENTS:: Open-ended and rating-scaled items. RESULTS:: Half of the proxies agreed with both of two rating-scaled items asking about different approaches to decision making-that is, agreeing that they would decide on the basis of how their relative would have decided and agreeing that they would decide on the basis of what they believed was in their relative's best interests. Narrative responses elaborated on themes within the following three major domains: Substituted judgment, best interests, and weighing substituted judgment and best interests. Substituted judgment was framed as honoring the patient's wishes and values. Best interests was described as a perceived duty to maintain quality of life and avoid burdens or risks. Weighing the two standards emerged as a challenging yet important, way of honoring wishes while maintaining quality of life. An unexpected theme was the attempt by alternative decision makers to discern their loved one's current versus premorbid research preferences. CONCLUSIONS:: Tensions exist between abstract ethical principles regarding decision-making "standards" and their translation into research decisions.

APJ1 and GRE3 Homologs Work in Concert to Allow Growth in Xylose in a Natural Saccharomyces Sensu Stricto Hybrid Yeast

Creating Saccharomyces yeasts capable of efficient fermentation of pentoses such as xylose remains a key challenge in the production of ethanol from lignocellulosic biomass. Metabolic engineering of industrial Saccharomyces cerevisiae strains has yielded xylose-fermenting strains, but these strains have not yet achieved industrial viability due largely to xylose fermentation being prohibitively slower than that of glucose. Recently, it has been shown that naturally occurring xylose-utilizing Saccharomyces species exist. Uncovering the genetic architecture of such strains will shed further light on xylose metabolism, suggesting additional engineering approaches or possibly even enabling the development of xylose-fermenting yeasts that are not genetically modified. We previously identified a hybrid yeast strain, the genome of which is largely Saccharomyces uvarum, which has the ability to grow on xylose as the sole carbon source. To circumvent the sterility of this hybrid strain, we developed a novel method to genetically characterize its xylose-utilization phenotype, using a tetraploid intermediate, followed by bulk segregant analysis in conjunction with high-throughput sequencing. We found that this strain's growth in xylose is governed by at least two genetic loci, within which we identified the responsible genes: one locus contains a known xylose-pathway gene, a novel homolog of the aldo-keto reductase gene GRE3, while a second locus contains a homolog of APJ1, which encodes a putative chaperone not previously connected to xylose metabolism. Our work demonstrates that the power of sequencing combined with bulk segregant analysis can also be applied to a non-genetically-tractable hybrid strain that contains a complex, polygenic trait, and identifies new avenues for metabolic engineering as well as for construction of non-genetically modified xylose-fermenting strains.

A New Method for Quantifying the Needling Component of Acupuncture Treatments

OBJECTIVES: The highly variable nature of acupuncture needling creates challenges to systematic research. The goal of this study was to test the feasibility of quantifying acupuncture needle manipulation using motion and force measurements. It was hypothesised that distinct needling styles and techniques would produce different needle motion and force patterns that could be quantified and differentiated from each other. METHODS: A new needling sensor tool (Acusensor) was used to record needling in real time as performed by six New England School of Acupuncture staff from the 'Chinese acupuncture' (style 1) and 'Japanese acupuncture' (style 2) programmes (three from each). Each faculty expert needled 12 points (6 bilateral locations) in 12 healthy human subjects using tonification (technique 1) and dispersal (technique 2). Parameters calculated from the raw needling data were displacement amplitude, displacement frequency, rotation amplitude, rotation frequency, force amplitude and torque amplitude. RESULTS: Data analysis revealed significant differences in the amplitude of displacement and rotation between needling performed by staff from two different acupuncture styles. Significant overall differences in the frequency of displacement between techniques 1 and 2 that were not dependent of the style of acupuncture being performed were also found. The relationships between displacement and rotation frequencies, as well as between displacement and force amplitudes showed considerable variability across individual acupuncturists and subjects. CONCLUSIONS: Needling motion and force parameters can be quantified in a treatment-like setting. Needling data can subsequently be analysed, providing an objective method for characterising needling in basic and clinical acupuncture research.

Medulloblastoma Stem Cells: Where Development and Cancer Cross Pathways

Brain tumors are the leading cause of childhood cancer mortality, with medulloblastoma (MB) representing the most frequent malignant tumor. The recent molecular classification of MB has reconceptualized the heterogeneity that exists within pathological subtypes by giving context to the role of key developmental signaling pathways in MB pathogenesis. The identification of cancer stem cell (CSC) populations, termed brain tumor-initiating cells (BTICs), in MB has provided novel cellular targets for the study of these aberrantly activated signaling pathways, namely, Sonic hedgehog (Shh) and Wingless (Wnt), along with the identification of novel BTIC self-renewal pathways. In this review, we discuss recent evidence for the presence of a MB stem cell that drives tumorigenesis in this malignant childhood tumor. We focus on evidence from cerebellar development, the recent identification of BTICs, the presence of activated developmental signaling pathways in MB, the role of epigenetic stem cell regulatory mechanisms, and how these developmental and epigenetic pathways may be targeted for novel therapeutic options.

Genome Stability of Lyme Disease Spirochetes: Comparative Genomics of Borrelia Burgdorferi Plasmids

Lyme disease is the most common tick-borne human illness in North America. In order to understand the molecular pathogenesis, natural diversity, population structure and epizootic spread of the North American Lyme agent, Borrelia burgdorferi sensu stricto, a much better understanding of the natural diversity of its genome will be required. Towards this end we present a comparative analysis of the nucleotide sequences of the numerous plasmids of B. burgdorferi isolates B31, N40, JD1 and 297. These strains were chosen because they include the three most commonly studied laboratory strains, and because they represent different major genetic lineages and so are informative regarding the genetic diversity and evolution of this organism. A unique feature of Borrelia genomes is that they carry a large number of linear and circular plasmids, and this work shows that strains N40, JD1, 297 and B31 carry related but non-identical sets of 16, 20, 19 and 21 plasmids, respectively, that comprise 33-40% of their genomes. We deduce that there are at least 28 plasmid compatibility types among the four strains. The B. burgdorferi ∼900 Kbp linear chromosomes are evolutionarily exceptionally stable, except for a short ≤20 Kbp plasmid-like section at the right end. A few of the plasmids, including the linear lp54 and circular cp26, are also very stable. We show here that the other plasmids, especially the linear ones, are considerably more variable. Nearly all of the linear plasmids have undergone one or more substantial inter-plasmid rearrangements since their last common ancestor. In spite of these rearrangements and differences in plasmid contents, the overall gene complement of the different isolates has remained relatively constant.

Relationship of Dopamine Type 2 Receptor Binding Potential with Fasting Neuroendocrine Hormones and Insulin Sensitivity in Human Obesity

OBJECTIVE Midbrain dopamine (DA) neurons, which are involved with reward and motivation, are modulated by hormones that regulate food intake (insulin, leptin, and acyl ghrelin [AG]). We hypothesized that these hormones are associated with deficits in DA signaling in obesity. RESEARCH DESIGN AND METHODS We assessed the relationships between fasting levels of insulin and leptin, and AG, BMI, and insulin sensitivity index (S(I)) with the availability of central DA type 2 receptor (D2R). We measured D2R availability using positron emission tomography and [(18)F]fallypride (radioligand that competes with endogenous DA) in lean (n = 8) and obese (n = 14) females. Fasting hormones were collected prior to scanning and S(I) was determined by modified oral glucose tolerance test. RESULTS Parametric image analyses revealed associations between each metabolic measure and D2R. The most extensive findings were negative associations of AG with clusters involving the striatum and inferior temporal cortices. Regional regression analyses also found extensive negative relationships between AG and D2R in the caudate, putamen, ventral striatum (VS), amygdala, and temporal lobes. S(I) was negatively associated with D2R in the VS, while insulin was not. In the caudate, BMI and leptin were positively associated with D2R availability. The direction of associations of leptin and AG with D2R availability are consistent with their opposite effects on DA levels (decreasing and increasing, respectively). After adjusting for BMI, AG maintained a significant relationship in the VS. We hypothesize that the increased D2R availability in obese subjects reflects relatively reduced DA levels competing with the radioligand. CONCLUSIONS Our findings provide evidence for an association between the neuroendocrine hormones and DA brain signaling in obese females.

Nerve Injuries Sustained During Warfare: Part I - Epidemiology

We describe 261 peripheral nerve injuries sustained in war by 100 consecutive service men and women injured in Iraq and Afghanistan. Their mean age was 26.5 years (18.1 to 42.6), the median interval between injury and first review was 4.2 months (mean 8.4 months (0.36 to 48.49)) and median follow-up was 28.4 months (mean 20.5 months (1.3 to 64.2)). The nerve lesions were predominantly focal prolonged conduction block/neurapraxia in 116 (45%), axonotmesis in 92 (35%) and neurotmesis in 53 (20%) and were evenly distributed between the upper and the lower limbs. Explosions accounted for 164 (63%): 213 (82%) nerve injuries were associated with open wounds. Two or more main nerves were injured in 70 patients. The ulnar, common peroneal and tibial nerves were most commonly injured. In 69 patients there was a vascular injury, fracture, or both at the level of the nerve lesion. Major tissue loss was present in 50 patients: amputation of at least one limb was needed in 18. A total of 36 patients continued in severe neuropathic pain. This paper outlines the methods used in the assessment of these injuries and provides information about the depth and distribution of the nerve lesions, their associated injuries and neuropathic pain syndromes.

Nerve Injuries Sustained During Warfare: Part II: Outcomes

The outcomes of 261 nerve injuries in 100 patients were graded good in 173 cases (66%), fair in 70 (26.8%) and poor in 18 (6.9%) at the final review (median 28.4 months (1.3 to 64.2)). The initial grades for the 42 sutures and graft were 11 good, 14 fair and 17 poor. After subsequent revision repairs in seven, neurolyses in 11 and free vascularised fasciocutaneous flaps in 11, the final grades were 15 good, 18 fair and nine poor. Pain was relieved in 30 of 36 patients by nerve repair, revision of repair or neurolysis, and flaps when indicated. The difference in outcome between penetrating missile wounds and those caused by explosions was not statistically significant; in the latter group the onset of recovery from focal conduction block was delayed (mean 4.7 months (2.5 to 10.2) vs 3.8 months (0.6 to 6); p = 0.0001). A total of 42 patients (47 lower limbs) presented with an insensate foot. By final review (mean 27.4 months (20 to 36)) plantar sensation was good in 26 limbs (55%), fair in 16 (34%) and poor in five (11%). Nine patients returned to full military duties, 18 to restricted duties, 30 to sedentary work, and 43 were discharged from military service. Effective rehabilitation must be early, integrated and vigorous. The responsible surgeons must be firmly embedded in the process, at times exerting leadership.

Kinetic Characterization of Newly Discovered Inhibitors of Various Constructs of Human T-cell Leukemia Virus-1 (HTLV-1) Protease and Their Effect on HTLV-1-infected Cells

BACKGROUND: Human T-cell leukemia virus-1 (HTLV-1) was the first identified human retrovirus and was shown to be associated with diseases such as adult T-cell leukemia lymphoma and tropical spastic paraparesis/HTLV-1 associated myelopathy. Retroviral proteases (PRs) are essential for viral replication by processing viral Gag and Gag-(Pro)-Pol polyproteins during maturation. Full-length HTLV-1 PR is 125 residues long; whether the C-terminal region is required for catalytic activity is still controversial. In this study, we characterized the effect of C-terminal amino acids of HTLV-1 PR for PR activity and examined the binding of compounds identified by in silico screening. One compound showed inhibition against the virus in infected cells. METHODS: Truncated (116-, 121- and 122-residue) forms of HTLV-1 PR were prepared and proteins from expression of the genes were purified. In silico screening was performed by docking small molecules into the active site of HTLV-1 PR. The kinetic constants k(cat), K(m), k(cat)/K(m) and inhibition constants K(i) for inhibitors identified by the computational screening were determined. Western blot and ELISA analyses were used to determine the effect of the most potent PR inhibitors on HTLV-1 protein processing in infected cells. RESULTS: The constructs showed similar catalytic efficiency constants (k(cat)/K(m)); thus HTLV-1 PR C-terminal amino acids are not essential for full activity. Computational screening revealed new PR inhibitors and some were shown to be inhibitory in enzyme assays. In HTLV-1-infected cells, one of the small molecules inhibited HTLV-1 gag cleavage and decreased the amount of HTLV-1 p19 produced in the cells. CONCLUSIONS: We have identified an HTLV-1 PR inhibitor that is biologically functional. Inhibitor screening will continue to develop possible drugs for therapy of HTLV-1 infection.

Genetic Versus Census Estimators of the Opportunity for Sexual Selection in the Wild

Abstract The existence of a direct link between intensity of sexual selection and mating-system type is widely accepted. However, the quantification of sexual selection has proven problematic. Several measures of sexual selection have been proposed, including the operational sex ratio (OSR), the breeding sex ratio (BSR), and the opportunity for sexual selection (I(mates)). For a wild population of pronghorn (Antilocapra americana), we calculated OSR and BSR. We estimated I(mates) from census data on the spatial and temporal distribution of receptive females in rut and from a multigenerational genetic pedigree. OSR and BSR indicated weak sexual selection on males, but census and pedigree I(mates) suggested stronger sexual selection on males than on females. OSR and BSR correlated with census but not pedigree estimates of I(mates), and census I(mates) did not correlate with pedigree estimates. This suggests that the behavioral mating system, as deduced from the spatial and temporal distribution of females, does not predict the genetic mating system of pronghorn. The differences we observed between estimators were primarily due to female mate sampling and choice and to the sex ratio. For most species, behavioral data are not perfectly accurate and therefore will be an insufficient alternative to using multigenerational pedigrees to quantify sexual selection.

Anti-NMDA Receptor Encephalitis. The Disorder, the Diagnosis and the Immunobiology

Anti-NMDAR encephalitis is a newly characterized syndrome with a progressive, predictable clinical course and the possibility of effective treatment. Accurate and timely diagnosis is critical to selection and implementation of treatments, and optimal patient outcomes. Outcomes are improved with early diagnosis via indirect immunofluorescence or cell-based assays, and the rapid and appropriate administration of immunosuppressant and anti-psychotic therapies. Three possible scenarios accounting for the immunopathogenesis of anti-NMDAR encephalitis are presented, with the most probable one being that of paraneoplastic autoimmunity. Future efforts in this disorder should focus on elucidating the mechanisms that contribute to initiation of this antibody response, as well as exploring the role of tumors, infectious triggers and immune-reactivation. Finally, accessible tools need to be developed that allow for reliable identification of specific antibody markers against synaptic proteins.

Connectedness, Social Support and Internalising Emotional and Behavioural Problems in Adolescents Displaced by the Chechen Conflict

The study investigated factors associated with internalising emotional and behavioural problems among adolescents displaced during the most recent Chechen conflict. A cross-sectional survey (N = 183) examined relationships between social support and connectedness with family, peers and community in relation to internalising problems. Levels of internalising were higher in displaced Chechen youth compared to published norms among non-referred youth in the United States and among Russian children not affected by conflict. Girls demonstrated higher problem scores compared to boys. Significant inverse correlations were observed between family, peer and community connectedness and internalising problems. In multivariate analyses, family connectedness was indicated as a significant predictor of internalising problems, independent of age, gender, housing status and other forms of support evaluated. Sub-analyses by gender indicated stronger protective relationships between family connectedness and internalising problems in boys. Results indicate that family connectedness is an important protective factor requiring further exploration by gender in war-affected adolescents.

"Hiding the Story": Indigenous Consumer Concerns About Communication Related to Chronic Disease in One Remote Region of Australia*

This paper reports on a collaborative qualitative study which explored education and communication practice related to chronic disease from the perspectives of Aboriginal people in a remote region of the Northern Territory, Australia, where the prevalence of chronic disease is extremely high. Most Yolngu (Aboriginal people of Northeast Arnhem Land) do not speak English as their first language and few health staff share the language and cultural background of their clients. Semi-structured interviews were conducted with Yolngu community members and health staff in their preferred language in small groups or individually, in an approach that was flexible and responsive to the concerns and priorities of Yolngu researchers and participants. As well, health education interactions were videotaped to facilitate more in-depth understanding of the strengths and challenges in communication (one video can be viewed at www.informahealthcare.com/ijslp ). An iterative and collaborative process of analysis, interpretation, and verification revealed that communication and education related to chronic disease is highly ineffective, restricting the extent to which Yolngu can make informed decisions in managing their health. Yolngu participants consistently stated that they wanted a detailed and direct explanation about causes and management of chronic disease from health staff, and rarely believed this had been provided, sometimes assuming that information about their health is deliberately withheld. These serious limitations in communication and education have extensive negative consequences for individuals, their families, and health services. These findings also have broader relevance to all areas of healthcare, including allied health services, which share similar challenges in achieving effective communication. Without addressing the profound and pervasive inadequacies in communication, other interventions designed to close the gap in Indigenous health are unlikely to succeed.

THE IMPACT OF EXPOSURE TO INTERPERSONAL VIOLENCE ON GENDER DIFFERENCES IN ADOLESCENT-ONSET MAJOR DEPRESSION: RESULTS FROM THE NATIONAL COMORBIDITY SURVEY REPLICATION (NCS-R)

BACKGROUND: Beginning in adolescence, females are at significantly higher risk for depression than males. Despite substantial efforts, gaps remain in our understanding of this disparity. This study tested whether gender differences in adolescent-onset depression arise because of female's greater exposure or sensitivity to violence. METHODS: Data came from 5,692 participants in the National Comorbidity Survey Replication. Trained interviewers collected data about major depression and participants' exposure to four types of interpersonal violence (physical abuse, sexual assault, rape, and witnessing violence) using a modified version of the Composite International Diagnostic Interview. We used discrete time survival analysis to investigate gender differences in the risk of adolescent onset depression. RESULTS: Of the entire sample, 5.7% met DSM-IV criteria for depression by age 18; 5.8% of the sample reported being physically abused, 11.7% sexually assaulted, 8.5% raped, and 13.2% witnessed violence by age 18. Females had 1.51 times higher odds of depression by age 18 than males. Exposure to all types of violence was associated with an increased odds of depression in both the past year and the years following exposure. Adjusting for exposure to violence partially attenuated the association between gender and depression, especially for sexual assault (odds ratio [OR] attenuated = 1.28; 15.23%) and rape (OR attenuated = 1.32; 12.59%). There was no evidence that females were more vulnerable to the effects of violence than males. DISCUSSION: Gender differences in depression are partly explained by females' higher likelihood of experiencing interpersonal violence. Reducing exposure to sexual assault and rape could therefore mitigate gender differences in depression.

Proteomics Clinical Applications: Serving the Field

Proteomics Clinical Applications Reviews 2012

Wellcome Trust: A Long-term Perspective

Understanding Public Health Informatics Competencies for Mid-tier Public Health Practitioners--a Web-based Survey

The abstract of this article was orally presented at American Public Health Association Annual Conference Health Informatics Information Technology Section in Philadelphia, PA, USA, November 2009. The literature suggests that there is a need for measuring public health informatics (PHI) competency to further understand whether current educational modules and modalities meet the needs of PHI practitioners and researchers to perform their jobs more effectively, particularly for mid-tier practitioners that constitute the majority of public health workers in the USA. The present study seeks to update current knowledge of the perceptions and experiences of PHI competencies proposed by the U.S. Council on Linkage in Public Health specifically for mid-tier PH practitioners and researchers. The results were collected and analyzed by using a Web-based survey (WBS) method administered among both practitioners and researchers. Researchers first compiled a draft list of candidate competency set by incorporating existing competency areas provided by: 1) the Council on Linkage; and by 2) those proposed by the USA's Centers for Disease Control CDC Public Health Informatics Work Group. Nine sets of competency statements with 120 competency items and demographic information of respondents were included in the WBS. The online survey instruments were pilot-tested accordingly to incorporate feedback from respondents of the pilot. Fifty-six subjects were recruited from PH experts who were: 1) members of the Health Informatics Information Technology (HIIT) group of American Public Health Association; and, 2) members from the Community of Science (COS) Website who were the first authors published in the PHI field from PubMed. The sample included diverse backgrounds of PHI workers. They expressed an increased need for training to improve their PHI competencies. Respondents agreed that four competency sets should be adequately represented, including Leadership and System Thinking Skills (82%), followed by Financial Planning and Management Skills (79%), Community Dimensions of Practice Skills (77%), and Policy Development/Program Planning Skills (63%). The findings parallel current literature indicating that there exists an expressed need for clarification of the public health practitioner's job-specific informatics competency. Findings of expressed needs for basic computer literacy training and community-based practice were consistent with those of the literature. Additional training and resources should be allocated to address the competency of leadership, management, community-based practice and policy advocacy skills for mid-tier public health practitioners to perform their jobs more effectively. Only when healthcare organizations properly identify PHI competency needs will public health practitioners likely improve their overall informatics skills while improving diversification for contribution across multiple settings.

Magnetic Resonance Imaging Findings in Spinal Tuberculosis: Comparison of HIV Positive and Negative Patients

There is an increasing incidence of Human immunodeficiency virus (HIV) and tuberculosis (TB) co-infection. This has led to an increasing number of atypical features on magnetic resonance imaging (MRI). We postulated that the type 4 hypersensitivity response causing granulomatous inflammation may be disrupted by the HIV resulting in less vertebral body destruction. This study compares the MRI features of spinal tuberculosis in HIV positive and negative patients.

Systemic Mycosis in a California Sea Lion (Zalophus Californianus) with Detection of Cystofilobasidiales DNA

A 6-yr-old, intact male California sea lion (Zalophus californianus) with a systemic mycosis died after 5 wk of antifungal drug therapy. Antemortem clinical findings included hind flipper swelling, ring-lesions on skin of the flippers, and dermal nodules that increased in size and number spreading from the hind flippers and ventral abdomen to the foreflippers and muzzle. Lesions were accompanied by severe lymphadenopathy and development of systemic clinical signs despite therapy using itraconazole and later voriconazole. Histopathologic evaluation of biopsies revealed granulomatous dermatitis due to infection by fungus-producing yeast cells in tissue. Isolation attempts, using biopsied skin and tissue samples collected at necropsy, failed to yield growth of a fungus producing yeast cells like those in histologic section. Consensus polymerase chain reaction (PCR) tests of biopsied skin for fungal DNA produced an amplicon having significant sequence identity with a Cystofilobasidiales, a fungus belonging to a subclade that includes several Cryptococcus spp. Histopathologic evaluation of necropsy tissues revealed a systemic mycosis with yeast cells disseminated throughout subcutis, lymph nodes, and viscera. Hepatic necrosis was identified associated with acute liver failure, possibly from the voriconazole administration. This is the first report documenting the clinical presentation, treatment, and pathologic findings of infection associated with Cystofilobasidiales in a marine mammal and serves to expand the understanding of mycoses in pinnipeds.

Longitudinal Changes in Semantic Categorization Performance After Symptomatic Remission from First-episode Psychosis: A 3-year Follow-up Study

Semantic categorization abnormalities have been observed in schizophrenia, but studies have rarely focused on the longitudinal trajectory. In this study, we consider semantic performance and the relationship with symptomatic changes during recovery from a first-episode of schizophrenia over a period of 3years. Thirty-seven first-episode patients with schizophrenia were compared to thirty-seven matched controls in a categorization task. Patients were assessed at first episode, after clinical stabilization, and annually for the subsequent 3years. In the task, participants indicated whether a word belonged to a given category. Each category contained words of varying degrees of semantic relatedness: typical, atypical, borderline, related-but-outside, and unrelated. Reaction times and proportion of 'yes' responses were analyzed. At first assessment, semantic categorization abnormalities were observed in first-episode patients. Patients assigned more semantically-dissimilar words to the categories than controls. As patients stabilized from acute states, their semantic categorization performance improved and then remained stable throughout the entire follow up period of 3years. Interestingly, semantic performance deficits, particularly a diminished typicality effect, correlated with negative symptoms in the initial episode, but not at stabilization when symptoms subsided. No significant associations between positive and negative symptoms, or pre-defined categorization measures were identified. The data demonstrated semantic memory abnormalities in first-episode schizophrenia. However, an improvement of semantic categorization performance was observed in stabilized schizophrenia patients. Overall, the data are suggestive of a state effect in semantic abnormalities rather than a trait effect. The correlation between degree of impairment and symptoms may explain previous inconsistent findings.

Association Between Pro- and Anti-inflammatory Cytokine Genes and a Symptom Cluster of Pain, Fatigue, Sleep Disturbance, and Depression

Because multiple symptoms associated with "sickness behavior" have a negative impact on functional status and quality of life, increased information on the mechanisms that underlie inter-individual variability in this symptom experience is needed. The purposes of this study were to determine: if distinct classes of individuals could be identified based on their experience with pain, fatigue, sleep disturbance, and depression; if these classes differed on demographic and clinical characteristics; and if variations in pro- and anti- inflammatory cytokine genes were associated with latent class membership. Self-report measures of pain, fatigue, sleep disturbance, and depression were completed by 168 oncology outpatients and 85 family caregivers (FCs). Using latent class profile analysis (LCPA), three relatively distinct classes were identified: those who reported low depression and low pain (83%), those who reported high depression and low pain (4.7%), and those who reported high levels of all four symptoms (12.3%). The minor allele of IL4 rs2243248 was associated with membership in the "All high" class along with younger age, being White, being a patient (versus a FC), having a lower functional status score, and having a higher number of comorbid conditions. Findings suggest that LPCA can be used to differentiate distinct phenotypes based on a symptom cluster associated with sickness behavior. Identification of distinct phenotypes provides new evidence for the role of IL4 in the modulation of a sickness behavior symptom cluster in oncology patients and their FCs.

Dandy Walker Variant with Treatment-resistant Bipolar Disorder

Evidence-based Path to Newborn Screening for Duchenne Muscular Dystrophy

Creatine kinase (CK) levels are increased on dried blood spots in newborns related to the birthing process. As a marker for newborn screening, CK in Duchenne muscular dystrophy (DMD) results in false-positive testing. In this report, we introduce a 2-tier system using the dried blood spot to first assess CK with follow-up DMD gene testing.

UK Ethnicity Data Collection for Healthcare Statistics: The South Asian Perspective

ABSTRACT: BACKGROUND: Ethnicity data collection has been proven to be important in health care but despite government initiatives remains incomplete and mostly un-validated in the UK. Accurate self-reported ethnicity data would enable experts to assess inequalities in health and access to services and help to ensure resources are targeted appropriately. The aim of this paper is to explore the reasons for the observed gap in ethnicity data by examining the perceptions and experiences of healthy South Asian volunteers. South Asians are the largest ethnic minority group accounting for 50% of all ethnic minorities in the UK 2001 census. METHODS: Five focus groups, conducted by trained facilitators in the native language of each group, recruited 36 South Asian volunteers from local community centres and places of worship. The topic guide focused on five key areas:1) general opinions on the collection of ethnicity, 2) experiences of providing ethnicity information, 3) categories used in practice, 4) opinions of other indicators of ethnicity e.g. language, religion and culture and 5) views on how should this information be collected. The translated transcripts were analysed using a qualitative thematic approach. RESULTS: The findings of this Cancer Research UK commissioned study revealed that participants felt that accurate recording of ethnicity data was important in healthcare with several stating the increased prevalence of certain diseases in minority ethnic groups as an appropriate justification to improve this data. The overwhelming majority raised no objections to providing this data when the purpose of data collection is fully explained. CONCLUSIONS: This study confirmed that the collection of patients' ethnicity data is deemed important by potential patients but there remains uncertainty and unease as to how the data may be used. A common theme running through the focus groups was the willingness to provide these data, strongly accompanied by a desire to have more information with regard to its use.

Hematopoietic Cell Transplant and Use of Massage for Improved Symptom Management: Results from a Pilot Randomized Control Trial

Background. Pediatric hematopoietic cell transplant (HCT) is a lifesaving treatment that often results in physical and psychological discomfort. An acupressure-massage intervention may improve symptom management in this setting. Methods. This randomized controlled pilot trial compared a combined massage-acupressure intervention to usual care. Children were offered three practitioner-provided sessions per week throughout hospitalization. Parents were trained to provide additional acupressure as needed. Symptoms were assessed using nurses' reports and two questionnaires, the behavioral affective and somatic experiences scale and the Peds quality of life cancer module. Results. We enrolled 23 children, ages 5 to 18. Children receiving the intervention reported fewer days of mucositis (Hedges' g effect size ES = 0.63), lower overall symptom burden (ES = 0.26), feeling less tired and run-down (ES = 0.86), having fewer moderate/severe symptoms of pain, nausea, and fatigue (ES = 0.62), and less pain (ES = 0.42). The intervention group showed trends toward increasing contentness/serenity (ES = +0.50) and decreasing depression (ES = -0.45), but not decreased anxiety (ES = +0.42). Differences were not statistically significant. Discussion. Feasibility of studying massage-acupressure was established in children undergoing HCT. Larger studies are needed to test the efficacy of such interventions in reducing HCT-associated symptoms in children.

Synthesis of the HCV Protease Inhibitor Vaniprevir (MK-7009) Using Ring-Closing Metathesis Strategy

A highly efficient synthesis of Vaniprevir (MK-7009) has been accomplished in nine linear steps and 55% overall yield. The key features of this synthesis include a cost-effective synthesis of the isoindoline subunit and efficient construction of the 20-membered macrocyclic core of Vaniprevir (MK-7009) utilizing ring-closing metathesis technology. A high-performing ring-closing metathesis protocol has been achieved by simultaneous slow addition of the ruthenium catalyst (0.2 mol %) and the diene substrate at a concentration of 0.13 M.

The Descriptive Epidemiology of Female Breast Cancer: An International Comparison of Screening, Incidence, Survival and Mortality

BACKGROUND: This paper presents the latest international descriptive epidemiological data for invasive breast cancer amongst women, including incidence, survival and mortality, as well as information on mammographic screening programmes. RESULTS: Almost 1.4 million women were diagnosed with breast cancer worldwide in 2008 and approximately 459,000 deaths were recorded. Incidence rates were much higher in more developed countries compared to less developed countries (71.7/100,000 and 29.3/100,000 respectively, adjusted to the World 2000 Standard Population) whereas the corresponding mortality rates were 17.1/100,000 and 11.8/100,000. Five-year relative survival estimates range from 12% in parts of Africa to almost 90% in the United States, Australia and Canada, with the differential linked to a combination of early detection, access to treatment services and cultural barriers. Observed improvements in breast cancer survival in more developed parts of the world over recent decades have been attributed to the introduction of population-based screening using mammography and the systemic use of adjuvant therapies. CONCLUSION: The future worldwide breast cancer burden will be strongly influenced by large predicted rises in incidence throughout parts of Asia due to an increasingly "westernised" lifestyle. Efforts are underway to reduce the global disparities in survival for women with breast cancer using cost-effective interventions.

Heterologous Expression and Purification of Arabidopsis Thaliana VIM1 Protein: In Vitro Evidence for Its Inability to Recognize Hydroxymethylcytosine, a Rare Base in Arabidopsis DNA

The discovery of 5-hydroxymethyl-cytosine (5hmC) in mammalian cells prompted us to look for this base in the DNA of Arabidopsis thaliana (thale cress), and to ask how well the Arabidopsis Variant in Methylation 1 (VIM1) protein, an essential factor in maintaining 5-cytosine methylation (5mC) homeostasis and epigenetic silencing in this plant, recognizes this novel base. We found that the DNA of Arabidopsis' leaves and flowers contain low levels of 5hmC. We also cloned and expressed in Escherichia coli full-length VIM1 protein, the archetypal member of the five Arabidopsis VIM gene family. Using in vitro binding assays, we observed that full-length VIM1 binds preferentially to hemi-methylated DNA with a single modified 5mCpG site; this result is consistent with its known role in preserving DNA methylation in vivo following DNA replication. However, when 5hmC replaces one or both cytosine residues at a palindromic CpG site, VIM1 binds with approximately ⩾10-fold lower affinity. These results suggest that 5hmC may contribute to VIM-mediated passive loss of cytosine methylation in vivo during Arabidopsis DNA replication.

A Nomenclature for Intestinal In Vitro Cultures

Many advances have been reported in the long term culture of intestinal mucosal cells in recent years. A significant number of publications have described new culture media, cell formations and growth patterns. Furthermore, it is now possible to study, e.g., the capabilities of isolated stem cells or the interactions between stem cells and mesenchyme. However, at the moment there is significant variation in the way these structures are described and named. A standardized nomenclature would benefit the ability to communicate and compare findings from different laboratories using the different culture systems. To address this issue, members of the NIH Intestinal Stem Cell Consortium herein propose a systematic nomenclature for in vitro cultures of the small and large intestine. We begin by describing the structures that are generated by preparative steps. We then define and describe structures produced in vitro, specifically: enterosphere, enteroid, reconstituted intestinal organoid, induced intestinal organoid, colonosphere, colonoid and colonic organoid.

Importance of Direct Patient Care in Advanced Pharmacy Practice Experiences

The Accreditation Council for Pharmacy Education issued revised standards (Standards 2007) for professional programs leading to the Doctor of Pharmacy degree in July 2007. The new standards require colleges and schools of pharmacy to provide pharmacy practice experiences that include direct interaction with diverse patient populations. These experiences are to take place in multiple practice environments (e.g., community, ambulatory care, acute care medicine, specialized practice areas) and must include face-to-face interactions between students and patients, and students and health care providers. In 2009, the American College of Clinical Pharmacy (ACCP) identified concerns among their members that training for some students during the fourth year of pharmacy curriculums are essentially observational experiences rather than encounters where students actively participate in direct patient care activities. These ACCP members also stated that there is a need to identify effective mechanisms for preceptors to balance patient care responsibilities with students' educational needs in order to fully prepare graduates for contemporary, patient-centered practice. The 2010 ACCP Educational Affairs Committee was charged to provide recommendations to more effectively foster the integration of pharmacy students into direct patient care activities during advanced pharmacy practice experiences (APPEs). In this commentary, the benefits to key stakeholders (pharmacy students, APPE preceptors, clerkship sites, health care institutions, academic pharmacy programs) of this approach are reviewed. Recommendations for implementation of direct patient care experiences are also provided, together with discussion of the practical issues associated with delivery of effective APPE. Examples of ambulatory care and acute care APPE models that successfully integrate pharmacy students into the delivery of direct patient care are described. Enabling students to engage in high-quality patient care experiences and to assume responsibility for drug therapy outcomes is achievable in a variety of practice settings. In our opinion, such an approach is mandatory if contemporary pharmacy education is to be successful in producing a skilled workforce capable of affecting drug therapy outcomes.

Mentoring Within Clinical Education

Enhanced Vasorelaxant Effects of the Endocannabinoid-like Mediator, Oleamide, in Hypertension

Oleamide is an endocannabinoid-like, fatty acid amide with structural similarities to anandamide. The cardiovascular effects of anandamide are enhanced in hypertension and we have now examined how hypertension affects responses to oleamide. Vasorelaxant responses to oleamide were significantly (P<0.001) enhanced in aortic rings from spontaneously hypertensive rats (SHRs), such that the maximal relaxation to oleamide was 40.3±3.5%, compared to 15.7±3.9% in normotensive Wistar Kyoto (WKY) controls. The augmented responses to oleamide in SHR arteries were unaffected by either inhibition of nitric oxide synthase (300μM l-NAME) or fatty acid amide hydrolase (1μM URB597) and independent of cannabinoid CB(1) receptors or the endothelium. The enhanced responses to oleamide were opposed by pre-treatment with capsaicin (such that R(max) was reduced to 9.8±1.5%) and this occurred independently of TRPV1 receptor and sensory nerve activity, as the TRPV1 antagonist capsazepine (1-5μM) and the cation channel inhibitor ruthenium red (10μM) had no effect on the responses to oleamide. However, inhibition of cyclooxygenase (10μM indomethacin) enhanced the responses in the WKY aortae, such that the responses were comparable to those in the SHR. The results suggest that the cyclooxygenase pathway has a role in modulating vasorelaxation caused by oleamide in normotensive aortae and that this is lost in hypertension, possibly as an adaptation to the increase in blood pressure.

Endoscopic Subtlety: Rest Assured Only When Biopsied

Effects of Compression on Muscle Tissue Oxygenation at the Onset of Exercise

The effects of compression on gastrocnemius medialis muscle oxygenation and hemodynamics during a short term dynamic exercise was investigated in a sample of fifteen male subjects (mean ± SD; age 25.8 ± 4.9 years; mass 70.6 ± 4.3 kg). Elastic compression sleeves were used to apply multiple levels of compression to the calf muscles during exercise and non-compressive garments were used for the control condition. Tissue hemoglobin oxygen saturation was measured as the relative "total oxygen index" (TOI) with a near infrared spectrometer (NIRS). The recovery of TOI during exercise was determined from the slope of oxygenation recovery in a non-occluded situation (NO). TOI recovery rate during the first two minutes of the exercise was 24% higher (p=0.042) for the compression condition than for the control condition. A significant correlation (r=0.61, p=0.012) between the level of compression and the tissue oxygenation recovery during exercise was observed. Muscle energy utilization was determined from the rate of decline of TOI immediately upon arterial occlusion (AO) during early exercise. Muscle energy utilization measured during the occluded situation was not significantly influenced by compression. Based on these results was concluded that compression induced changes in tissue blood flow/perfusion appear to result in improved oxygenation during short-term exercise. Assuming that increased muscle oxygen availability positively influences performance, compression of muscles may enhance performance especially in sports that require repeated short bouts of exercise.

Design and Synthesis of Potent, Orally Efficacious Hydroxyethylamine Derived β-Site Amyloid Precursor Protein Cleaving Enzyme (BACE1) Inhibitors

We have previously shown that hydroxyethylamines can be potent inhibitors of the BACE1 enzyme and that the generation of BACE1 inhibitors with CYP 3A4 inhibitory activities in this scaffold affords compounds (e.g., 1) with sufficient bioavailability and pharmacokinetic profiles to reduce central amyloid-β peptide (Aβ) levels in wild-type rats following oral dosing. In this article, we describe further modifications of the P1-phenyl ring of the hydroxyethylamine series to afford potent, dual BACE1/CYP 3A4 inhibitors which demonstrate improved penetration into the CNS. Several of these compounds caused robust reduction of Aβ levels in rat CSF and brain following oral dosing, and compound 37 exhibited an improved cardiovascular safety profile relative to 1.

Different Selective Pressures Lead to Different Genomic Outcomes As Newly-formed Hybrid Yeasts Evolve

ABSTRACT: BACKGROUND: Interspecific hybridization occurs in every eukaryotic kingdom. While hybrid progeny are frequently at a selective disadvantage, in some instances their increased genome size and complexity may result in greater stress resistance than their ancestors, which can be adaptively advantageous at the edges of their ancestors' ranges. While this phenomenon has been repeatedly documented in the field, the response of hybrid populations to long-term selection has not often been explored in the lab. To fill this knowledge gap we crossed the two most distantly related members of the Saccharomyces sensu stricto group, S. cerevisiae and S. uvarum, and established a mixed population of homoploid and aneuploid hybrids to study how different types of selection impact hybrid genome structure. RESULTS: As temperature was raised incrementally from 31degreesC to 46.5degreesC over 500 generations of continuous culture, selection favored loss of the S. uvarum genome, although the kinetics of genome loss differed among independent replicates. Temperature-selected isolates exhibited greater inherent and induced thermal tolerance than parental species and founding hybrids, and also exhibited ethanol resistance. In contrast, as exogenous ethanol was increased from 0% to 14% over 500 generations of continuous culture, selection favored euploid S. cerevisiae x S. uvarum hybrids. Ethanol-selected isolates were more ethanol tolerant than S. uvarum and one of the founding hybrids, but did not exhibit resistance to temperature stress. Relative to parental and founding hybrids, temperature-selected strains showed heritable differences in cell wall structure in the forms of increased resistance to zymolyase digestion and Micafungin, which targets cell wall biosynthesis. CONCLUSIONS: This is the first study to show experimentally that the genomic fate of newly-formed interspecific hybrids depends on the type of selection they encounter during the course of evolution, underscoring the importance of the ecological theatre in determining the outcome of the evolutionary play.

High-Performance Sodium-Ion Pseudocapacitors Based on Hierarchically Porous Nanowire Composites

Electrical energy storage plays an increasingly important role in modern society. Current energy storage methods are highly dependent on lithium-ion energy storage devices, and the expanded use of these technologies is likely to affect existing lithium reserves. The abundance of sodium makes Na-ion-based devices very attractive as an alternative, sustainable energy storage system. However, electrodes based on transition-metal oxides often show slow kinetics and poor cycling stability, limiting their use as Na-ion-based energy storage devices. The present paper details a new direction for electrode architectures for Na-ion storage. Using a simple hydrothermal process, we synthesized interpenetrating porous networks consisting of layer-structured V(2)O(5) nanowires and carbon nanotubes (CNTs). This type of architecture provides facile sodium insertion/extraction and fast electron transfer, enabling the fabrication of high-performance Na-ion pseudocapacitors with an organic electrolyte. Hybrid asymmetric capacitors incorporating the V(2)O(5)/CNT nanowire composites as the anode operated at a maximum voltage of 2.8 V and delivered a maximum energy of ∼40 Wh kg(-1), which is comparable to Li-ion-based asymmetric capacitors. The availability of capacitive storage based on Na-ion systems is an attractive, cost-effective alternative to Li-ion systems.

PROTEOMICS Reviews 2012

Intervention Research, Randomised Trials and Knowledge for Decision-making

Differential Coloring Reveals That Plastids Do Not Form Networks for Exchanging Macromolecules

Stroma-filled tubules named stromules are sporadic extensions of plastids. Earlier, photobleaching was used to demonstrate fluorescent protein diffusion between already interconnected plastids and formed the basis for suggesting that all plastids are able to form networks for exchanging macromolecules. However, a critical appraisal of literature shows that this conjecture is not supported by unequivocal experimental evidence. Here, using photoconvertible mEosFP, we created color differences between similar organelles that enabled us to distinguish clearly between organelle fusion and nonfusion events. Individual plastids, despite conveying a strong impression of interactivity and fusion, maintained well-defined boundaries and did not exchange fluorescent proteins. Moreover, the high pleomorphy of etioplasts from dark-grown seedlings, leucoplasts from roots, and assorted plastids in the accumulation and replication of chloroplasts5 (arc5), arc6, and phosphoglucomutase1 mutants of Arabidopsis thaliana suggested that a single plastid unit might be easily mistaken for interconnected plastids. Our observations provide succinct evidence to refute the long-standing dogma of interplastid connectivity. The ability to create and maintain a large number of unique biochemical factories in the form of singular plastids might be a key feature underlying the versatility of green plants as it provides increased internal diversity for them to combat a wide range of environmental fluctuations and stresses.

Massage for Children Undergoing Hematopoietic Cell Transplantation: a Qualitative Report

Background. No in-depth qualitative research exists about the effects of therapeutic massage with children hospitalized to undergo hematopoietic cell transplantation (HCT). The objective of this study is to describe parent caregivers' experience of the effects of massage/acupressure for their children undergoing HCT. Methods. We conducted a qualitative analysis of open-ended interviews with 15 parents of children in the intervention arm of a massage/acupressure trial. Children received both practitioner and parent-provided massage/acupressure. Results. Parents reported that their child experienced relief from pain and nausea, relaxation, and greater ease falling asleep. They also reported increased caregiver competence and closeness with their child as a result of learning and performing massage/acupressure. Parents supported a semistandardized massage protocol. Conclusion. Massage/acupressure may support symptom relief and promote relaxation and sleep among pediatric HCT patients if administered with attention to individual patients' needs and hospital routines and may relieve stress among parents, improve caregiver competence, and enhance the sense of connection between parent and child.

Reductively Responsive SiRNA-Conjugated Hydrogel Nanoparticles for Gene Silencing

A critical need still remains for effective delivery of RNA interference (RNAi) therapeutics to target tissues and cells. Self-assembled lipid- and polymer-based systems have been most extensively explored for transfection with small interfering RNA (siRNA) in liver and cancer therapies. Safety and compatibility of materials implemented in delivery systems must be ensured to maximize therapeutic indices. Hydrogel nanoparticles of defined dimensions and compositions, prepared via a particle molding process that is a unique off-shoot of soft lithography known as particle replication in nonwetting templates (PRINT), were explored in these studies as delivery vectors. Initially, siRNA was encapsulated in particles through electrostatic association and physical entrapment. Dose-dependent gene silencing was elicited by PEGylated hydrogels at low siRNA doses without cytotoxicity. To prevent disassociation of cargo from particles after systemic administration or during postfabrication processing for surface functionalization, a polymerizable siRNA pro-drug conjugate with a degradable, disulfide linkage was prepared. Triggered release of siRNA from the pro-drug hydrogels was observed under a reducing environment while cargo retention and integrity were maintained under physiological conditions. Gene silencing efficiency and cytocompatibility were optimized by screening the amine content of the particles. When appropriate control siRNA cargos were loaded into hydrogels, gene knockdown was only encountered for hydrogels containing releasable, target-specific siRNAs, accompanied by minimal cell death. Further investigation into shape, size, and surface decoration of siRNA-conjugated hydrogels should enable efficacious targeted in vivo RNAi therapies.

Universal Newborn Screening: Knowledge, Attitudes, and Satisfaction Among Public Health Professionals

Assess knowledge, attitude, and satisfaction with the newborn screening (NBS) system among pediatric public health leaders in the state of Florida.

A Novel Methodology for 3D Deformable Dosimetry

Purpose: Interfraction and intrafraction variation in anatomic structures is a significant challenge in contemporary radiotherapy. The objective of this work is to develop a novel tool for deformable structure dosimetry, using a tissue-equivalent deformable gel dosimeter that can reproducibly simulate targets subject to deformation. This will enable direct measurement of integrated doses delivered in different deformation states, and the verification of dose deforming algorithms.Methods: A modified version of the nPAG polymer gel has been used as a deformable 3D dosimeter and phantom to investigate doses delivered to deforming tissue-equivalent geometry. The deformable gel (DEFGEL) dosimeter/phantom is comprised of polymer gel in a latex membrane, moulded (in this case) into a cylindrical geometry, and deformed with an acrylic compressor. Fifteen aluminium fiducial markers (FM) were implanted into DEFGEL phantoms and the reproducibility of deformation was determined via multiple computed tomography (CT) scans in deformed and nondeformed states before and after multiple (up to 150) deformations. Dose was delivered to the DEFGEL phantom in three arrangements: (i) without deformation, (ii) with deformation, and (iii) cumulative exposures with and without deformation, i.e., dose integration. Irradiations included both square field and a stereotactic multiple dynamic arc treatment adapted from a patient plan. Doses delivered to the DEFGEL phantom were read out using cone beam optical CT.Results: Reproducibility was verified by observation of interscan shifts of FM locations (as determined via CT), measured from an absolute reference point and in terms of inter-FM distance. The majority (76%) of points exhibited zero shift, with others shifting by one pixel size consistent with setup error as confirmed with a control sample. Comparison of dose profiles and 2D isodose distributions from the three arrangements illustrated complex spatial redistribution of dose in all three dimensions occurring as a result of the change in shape of the target between irradiations, even for a relatively simple deformation. Discrepancies of up to 30% of the maximum dose were evident from dose difference maps for three orthogonal planes taken through the isocenter of a stereotactic field.Conclusions: This paper describes the first use of a tissue-equivalent, 3D dose-integrating deformable phantom that yields integrated or redistributed dosimetric information. The proposed methodology readily yields three-dimensional (3D) dosimetric data from radiation delivery to the DEFGEL phantom in deformed and undeformed states. The impacts of deformation on dose distributions were readily seen in the isodose contours and line profiles from the three arrangements. It is demonstrated that the system is potentially capable of reproducibly emulating the physical deformation of an organ, and therefore can be used to evaluate absorbed doses to deformable targets and organs at risk in three dimensions and to validate deformation algorithms applied to dose distributions.

Sindbis Virus Infectivity Improves During the Course of Infection in Both Mammalian and Mosquito Cells

Alphaviruses are enveloped, single-stranded positive sense RNA viruses that are transmitted by an arthropod vector to a wide host range, including avian and mammalian species. Arthropods and vertebrates have different cellular environments and this may cause the different cellular pathologies that are observed between the invertebrate vector and vertebrate hosts in both whole organisms and cultured cell lines. In this report, we used Sindbis virus and examined mosquito and mammalian cell lines for their ability to produce progeny virus particles. Total particles produced, viral titers, and overall infectivity (or the ratio of total particles-to-infectious particles) was investigated. Our results show (1) Sindbis infectivity is more a function of the host cell used in titering the virus rather than the cell line used to produce the virus, (2) the number of total and infectious particles produced is cell line dependent, and (3) the infectivity of released virus particles improves during the course of infection in both cells that have cytolytic infections and persistent infections.

Effects of Drop-out on Efficacy Estimates in Five Cochrane Reviews of Popular Antipsychotics for Schizophrenia

Hutton P, Morrison AP, Yung AR, Taylor PJ, French P, Dunn G. Effects of drop-out on efficacy estimates in five Cochrane reviews of popular antipsychotics for schizophrenia. Objective:  Our aim was to find out how Cochrane reviews of five popular or frequently prescribed second-generation antipsychotics in the UK (olanzapine, risperidone, quetiapine, amisulpride and aripiprazole) approached the problem of high drop-out in placebo-controlled trials. Method:  We examined the following: (i) whether reviews included data from studies with a level of drop-out exceeding their stated exclusion criterion; (ii) the level of missing data each efficacy outcome in each review relied upon; and (iii) impact of excluding studies with high drop-out. Results:  All reviews included data they stated they would exclude because of unacceptable levels of attrition, four (risperidone, olanzapine, amisulpride, aripiprazole) without clear acknowledgement or justification. Several reviews also excluded data from a number of relatively low-attrition studies because of missing standard deviations. Conclusion:  Cochrane reviews of five popular antipsychotics for schizophrenia misrepresented the available evidence on their efficacy. The impact of including high-attrition studies was difficult to quantify because of the exclusion of relevant low-attrition studies. Further analysis of the efficacy of these drugs in studies with acceptable rates of attrition is required. To reduce the problem of high attrition, trialists should gather follow-up data from people who leave the double-blind process early.

Australasian College for Emergency Medicine Trainee Research Project Bar is Not Too High

Heterogeneity of High Grade Cervical Intraepithelial Neoplasia Related to HPV16: Implications for Natural History and Management

Factors associated with progression from cervical intraepithelial neoplasias (CIN) grade 2 and 3 to invasive cancer are not well understood; most CIN2 and CIN3 regress spontaneously. Among carcinogenic HPV types, infections with HPV16 have the highest risk of progressing to cancer. We evaluated the heterogeneity of risk factors, lesion size, colposcopic impression, and colposcopic biopsy results in relation to HPV16 status among 627 women with CIN2 or CIN3 in women referred to colposcopy at the University of Oklahoma. Loop excision specimens were evaluated in 12 radial segments to estimate lesion size. The mean age at CIN3 was 27.7 years for HPV16-positive women (n=225) and 33.6 years for HPV16-negative women (n=104), respectively. The average lesion size did not differ by HPV16 status (p=0.83). Among HPV16-positive women with CIN3, lesions were significantly larger in women 30 years and older (p=0.03). Colposcopic impression was worse in women with HPV16 infections (p=0.009), but the detection of CIN3 at the preceding biopsy was not improved in HPV16-positive women. CIN3 is detected at the same lesion size, but at much younger age in women with HPV16 infections, suggesting faster growth. CIN2 lesion size in women without HPV16 peaks below 30 years and then decreases, suggesting frequent regression, while HPV16-related CIN2 is more likely to persist. Lesion size seems to be the most important determinant of colposcopy and biopsy performance. Genotyping for HPV16 in cervical cancer screening can improve risk stratification, but may pose challenges to finding small lesions in colposcopy. © 2012 Wiley-Liss, Inc.

Who Will Cover the Cost of Undocumented Immigrant Trauma Care?

: Health care reform under the "Patient Protection and Affordable Care Act" (PPACA) will lead to changes in reimbursement. Although this legislation provides a mechanism for uninsured Americans to obtain coverage, it excludes undocumented immigrants (UDI). Reimbursement for UDIs comes from the disproportionate share hospital (DSH) program and was previously supported by Section-1011 of the 2003 Medicare Modernization Act (S1011). The PPACA details a cut of DSH funds starting in 2014. This could impose a significant financial burden on trauma centers.

Short- and Long-term Dynamic Responses of the Metabolic Network and Gene Expression in Yeast to a Transient Change in the Nutrient Environment

Quantitative data on the dynamic changes in the transcriptome and the metabolome of yeast in response to an impulse-like perturbation in nutrient availability was integrated with the metabolic pathway information in order to elucidate the long-term dynamic re-organization of the cells. This study revealed that, in addition to the dynamic re-organization of the de novo biosynthetic pathways, salvage pathways were also re-organized in a time-dependent manner upon catabolite repression. The transcriptional and the metabolic responses observed for nitrogen catabolite repression were not as severe as those observed for carbon catabolite repression. Selective up- or down regulation of a single member of a paralogous gene pair during the response to the relaxation from nutritional limitation was identified indicating a differentiation of functions among paralogs. Our study highlighted the role of inosine accumulation and recycling in energy homeostasis and indicated possible bottlenecks in the process.

Early Detection and Intervention Evaluation for People at Risk of Psychosis: Multisite Randomised Controlled Trial

To determine whether cognitive therapy is effective in preventing the worsening of emerging psychotic symptoms experienced by help seeking young people deemed to be at risk for serious conditions such as schizophrenia.

Neighborhood Influences on Perceived Social Support Among Parents: Findings from the Project on Human Development in Chicago Neighborhoods

Social support is frequently linked to positive parenting behavior. Similarly, studies increasingly show a link between neighborhood residential environment and positive parenting behavior. However, less is known about how the residential environment influences parental social support. To address this gap, we examine the relationship between neighborhood concentrated disadvantage and collective efficacy and the level and change in parental caregiver perceptions of non-familial social support.

Regional Cerebral Blood Flow and FDG Uptake in Asymptomatic HIV-1 Men

Despite advances in the treatment of patients with human immunodeficiency virus (HIV), HIV-associated neurocognitive disorder occurs in 15-50% of HIV-infected individuals, and may become more apparent as ageing advances. In the present study we investigated regional cerebral blood flow (rCBF) and regional cerebral metabolic rate of glucose uptake (rCMRglc) in medically and psychiatrically stable HIV-1-infected participants in two age-groups. Positron emission tomography (PET) and magnetic resonance imaging (MRI)-based arterial spin labeling (ASL) were used to measure rCMRglc and rCBF, respectively, in 35 HIV-infected participants and 37 HIV-negative matched controls. All participants were currently asymptomatic with undetectable HIV-1 viral loads, without medical or psychiatric comorbidity, alcohol or substance misuse, stable on medication for at least 6 months before enrolment in the study. We found significant age effects on both ASL and PET with reduced rCBF and rCMRglc in related frontal brain regions, and consistent, although small, reductions in rCBF and rCMRglc in the anterior cingulate cortex (ACC) in HIV, a finding of potential clinical significance. There was no significant interaction between HIV status and the ageing process, and no significant HIV-related changes elsewhere in the brain on PET or ASL. This is the first paper to combine evidence from ASL and PET method in HIV participants. These finding provide evidence of crossvalidity between the two techniques, both in ageing and a clinical condition (HIV). Hum Brain Mapp , 2012. © 2012 Wiley Periodicals, Inc.

THORACOSCORE Fails to Predict Complications Following Elective Lung Resection

Thoracoscore mortality risk model has been incorporated into the British Thoracic Society guidelines on the radical management of patients with lung cancer. The discriminative and predictive ability to predict mortality and postoperative pulmonary complications (PPC) in this group of patients is uncertain.A prospective observational study was carried out on all patients following lung resection via thoracotomy in a regional thoracic centre over 42 months.128 of 703 subjects developed a PPC. 16 patients died in hospital (2%). In a logistic regression analysis the Thoracoscore was not a significant predictor of mortality (p=0.11; odds ratio (OR) =1.07, confidence interval (CI) 0.99-1.17) but was significant predictor of PPC (p=0.002; OR=1.08, CI 1.03-1.13). However, the area under the receiver operator characteristic (ROC) curve for the Thoracoscore was 0.68 (CI 0.56-0.80) for predicting mortality and 0.64 (CI 0.59-0.69) for PPC, indicating limited discriminative ability.In a logistic regression analysis, another risk model the European Society's Objective Score (ESOS) was predictive of mortality (p=0.006; OR =1.43, CI 1.11-1.83) and PPC (p<0.0001; OR =1.48, CI 1.30-1.68).Thoracoscore may have poor discriminative and predictive ability for mortality and PPC following elective lung resection.

Diet and Scavenging Habits of the Smooth Skate Dipturus Innominatus

The diet of smooth skate Dipturus innominatus was determined from examination of stomach contents of 321 specimens of 29·3-152·0 cm pelvic length, sampled from research and commercial trawlers at depths of 231-789 m on Chatham Rise, New Zealand. The diet was dominated by the benthic decapods Metanephrops challengeri and Munida gracilis, the natant decapod Campylonotus rathbunae and fishes from 17 families, of which hoki Macruronus novaezelandiae, sea perch Helicolenus barathri, various Macrouridae and a variety of discarded fishes were the most important. Multivariate analyses indicated the best predictors of diet variability were D. innominatus length and a spatial model. The diet of small D. innominatus was predominantly small crustaceans, with larger crustaceans, fishes and then scavenged discarded fishes increasing in importance as D. innominatus got larger. Scavenged discards were obvious as fish heads or tails only, or skeletal remains after filleting, often from pelagic species. Demersal fish prey were most frequent on the south and west Chatham Rise, in areas where commercial fishing was most active. Dipturus innominatus are highly vulnerable to overfishing, but discarding practices by commercial fishing vessels may provide a positive feedback to populations through improved scavenging opportunities.

Connexin-43 in the Osteogenic BM Niche Regulates Its Cellular Composition and the Bidirectional Traffic of Hematopoietic Stem Cells and Progenitors

Connexin-43 (Cx43), a gap junction protein involved in control of cell proliferation, differentiation and migration, has been suggested to have a role in hematopoiesis. Cx43 is highly expressed in osteoblasts and osteogenic progenitors (OB/P). To elucidate the biologic function of Cx43 in the hematopoietic microenvironment (HM) and its influence in HSC activity, we have studied the hematopoietic function in an in vivo model of constitutive deficiency of Cx43 in OB/P. The deficiency of Cx43 in OB/P cells does not impair the steady state hematopoiesis, but disrupts the directional trafficking of HSC/P between the BM and PB. OB/P Cx43 is a crucial positive regulator of trans-stromal migration and homing of both HSC and progenitors in an irradiated microenvironment. However, OB/P Cx43 deficiency in non-myeloablated animals does not result in a homing defect but induces increased endosteal lodging and decreased mobilization of HSC/P associated with proliferation and expansion of Cxcl12-secreting mesenchymal/osteolineage cells in the BM HM in vivo. Cx43 controls the cellular content of the BM osteogenic microenvironment and is required for homing of HSC/P in myeloablated animals.

Differences in Cognitive and Emotional Processes Between Persecutory and Grandiose Delusions

Background:Cognitive models propose that cognitive and emotional processes, in the context of anomalies of experience, lead to and maintain delusions. No large-scale studies have investigated whether persecutory and grandiose delusions reflect differing contributions of reasoning and affective processes. This is complicated by their frequent cooccurrence in schizophrenia. We hypothesized that persecutory and grandiose subtypes would differ significantly in their associations with psychological processes.Methods:Participants were the 301 patients from the Psychological Prevention of Relapse in Psychosis Trial (ISRCTN83557988). Persecutory delusions were present in 192 participants, and grandiose delusions were present in 97, while 58 were rated as having delusions both of persecution and grandiosity. Measures of emotional and reasoning processes, at baseline only, were employed.Results:A bivariate response model was used. Negative self-evaluations and depression and anxiety predicted a significantly increased chance of persecutory delusions whereas grandiose delusions were predicted by less negative self-evaluations and lower anxiety and depression, along with higher positive self and positive other evaluations. Reasoning biases were common in the whole group and in categorically defined subgroups with only persecutory delusions and only grandiose delusions; however, jumping to conclusions, and belief flexibility were significantly different in the 2 groups, the grandiose group having a higher likelihood of showing a reasoning bias than the persecutory group.Conclusion:The significant differences in the processes associated with these 2 delusion subtypes have implications for etiology and for the development of targeted treatment strategies.

Serendipity in Discovery of Proteasome Inhibitors

Among its various catalytic activities, the 'chymotrypsin-like' activity of the proteasome, a large multicatalytic proteinase complex has emerged as the focus of drug discovery efforts in cancer therapy. Herein, a series of first generation (2S, 3R)-2-amino-3-hydroxybutyric acid derived proteasome inhibitors that were discovered serendipitously en route to original goal of generating a series of sterically constrained oxazoline derivatives has been reported.

Elastic Deformations of the Rotary Double Motor of Single C Synthases Detected in Real Time by Förster Resonance Energy Transfer

Elastic conformational changes of the protein backbone are essential for catalytic activities of enzymes. To follow relative movements within the protein, Förster-type resonance energy transfer (FRET) between two specifically attached fluorophores can be applied. FRET provides a precise ruler between 3 and 8nm with subnanometer resolution. Corresponding submillisecond time resolution is sufficient to identify conformational changes in FRET time trajectories. Analyzing single enzymes circumvents the need for synchronization of various conformations. F(O)F(1)-ATP synthase is a rotary double motor which catalyzes the synthesis of adenosine triphosphate (ATP). A proton-driven 10-stepped rotary F(O) motor in the Escherichia coli enzyme is connected to a 3-stepped F(1) motor, where ATP is synthesized. To operate the double motor with a mismatch of step sizes smoothly, elastic deformations within the rotor parts have been proposed by W. Junge and coworkers. Here we extend a single-molecule FRET approach to observe both rotary motors simultaneously in individual F(O)F(1)-ATP synthases at work. We labeled this enzyme with two fluorophores specifically, that is, on the ε- and c-subunits of the two rotors. Alternating laser excitation was used to select the FRET-labeled enzymes. FRET changes indicated associated transient twisting within the rotors of single enzyme molecules during ATP hydrolysis and ATP synthesis. Supported by Monte Carlo simulations of the FRET experiments, these studies reveal that the rotor twisting is greater than 36° and is largely suppressed in the presence of the rotation inhibitor DCCD. This article is part of a Special Issue entitled: 17th European Bioenergetics Conference (EBEC 2012).

Evaluation of Current Molecular Approaches for Genotyping of Campylobacter Jejuni Strains

Abstract Campylobacter jejuni has been recognized as the most common bacterial cause of gastroenteritis worldwide, in both developed and developing countries, since the late 1970s. A number of genotyping schemes have been developed to identify the sources and route of transmission of these foodborne pathogens so that proper control measures can be developed. In this review, we provide current genotypic schemes developed for Campylobacter spp. (particularly C. jejuni) over the last decades, along with an evaluation of the strength and weakness of these techniques and their applications.

Emerging Insights into the Molecular and Cellular Basis of Glioblastoma

Glioblastoma is both the most common and lethal primary malignant brain tumor. Extensive multiplatform genomic characterization has provided a higher-resolution picture of the molecular alterations underlying this disease. These studies provide the emerging view that "glioblastoma" represents several histologically similar yet molecularly heterogeneous diseases, which influences taxonomic classification systems, prognosis, and therapeutic decisions.

Enhancing Angiogenesis Alleviates Hypoxia and Improves Engraftment of Enteric Cells in Polycaprolactone Scaffolds

We examined whether expediting angiogenesis in porous polycaprolactone (PCL) scaffolds could reduce hypoxia and consequently improve the survival of transplanted enteric cells. To accelerate angiogenesis, we delivered vascular endothelial growth factor (VEGF) using PCL scaffolds with surface crosslinked heparin. The fabrication and characterization of scaffolds has been reported in our previous study. Enteric cells, isolated from intestinal tissue of neonatal mice and expanded in vitro for 10 days, exhibited high expression levels for contractile protein α-smooth muscle actin and desmin. The cultured enteric cells were seeded in scaffolds and were implanted subcutaneously in immunodeficient mice for 7 and 14 days. At day 7, the heparin-modified PCL scaffolds with VEGF exhibited significantly increased angiogenesis and engraftment of enteric cells, with a simultaneous reduction in hypoxia. At day 14, the blood vessels grew across the entire thickness of the scaffold and resulted in a significantly diminished hypoxic environment; however, the transplanted cell density did not increase further. In conclusion, the enhancement of angiogenesis reduced cellular hypoxia and improved the engraftment of enteric cells. Copyright © 2012 John Wiley & Sons, Ltd.

Serial Assessment of Lymphocytes and Apoptosis in the Prostate During Coordinated Intraprostatic Dendritic Cell Injection and Radiotherapy

Local radiotherapy plus intratumoral syngeneic dendritic cell injection can mediate apoptosis/cell death and immunological tumor eradication in murine models. A novel method of coordinated intraprostatic, autologous dendritic cell injection together with radiation therapy was prospectively evaluated in five HLA-A2(+) subjects with high-risk, localized prostate cancer, using androgen suppression, 45 Gy external beam radiation therapy in 25 fractions over 5 weeks, dendritic cell injections after fractions 5, 15 and 25 and then interstitial radioactive seed placement. Serial prostate biopsies before and during treatment showed increased apoptotic cells and parenchymal distribution of CD8(+) cells. CD8(+) T-cell responses to test peptides were assessed using an enzyme-linked immunosorbent spot IFN-γ production assay, demonstrating some prostate cancer-specific protein-derived peptides associated with increased titer. In conclusion, the technique was feasible and well-tolerated and specific immune responses were observable. Future trials could further test the utility of this approach and improve on temporal coordination of intratumoral dendritic cell introduction with particular timelines of therapy-induced apoptosis.

Bacteriology of the Paranasal Sinuses in Pediatric Cystic Fibrosis Patients

OBJECTIVES: To review the characteristic microbiology of the paranasal sinuses in patients with cystic fibrosis who undergo endoscopic sinus surgery. To examine the subtypes of organisms cultured from the maxillary sinuses and determine their sensitivity to antibiotic therapy. STUDY DESIGN: Retrospective chart review. SETTING: Tertiary care children's hospital. METHODS: Sinus cultures were obtained from 51 patients with cystic fibrosis during endoscopic sinus procedures between 2000 and 2004 at a tertiary care children's hospital. A retrospective chart review was undertaken to obtain culture and sensitivity data of the sinus contents. RESULTS: The most common bacteria isolated was Staphylococcus aureus (71%), followed by Pseudomonas aeruginosa (PSA) (27%), Haemophilus influenzae (21%), Staphylococcus non-aureus (16%) and Streptococcus viridans (12%). Streptococcus pneumoniae and Moraxella catarrhalis were rarely isolated (2% and 0% respectively). Twenty-nine percent of the patients with cultures positive for PSA were of the mucoid variant. Only one patient had culture positive Escherichia coli. Antibiotic resistance among the more common organisms cultured from the sinus samples is also listed. CONCLUSION: Staph. aureus is the most common isolate in the sinuses of this pediatric CF population followed by P. aeruginosa and H. influenzae. Although many isolates are pansensitive, some isolates are panresistant.

Mouse Estrous Cycle Identification Tool and Images

The efficiency of producing timed pregnant or pseudopregnant mice can be increased by identifying those in proestrus or estrus. Visual observation of the vagina is the quickest method, requires no special equipment, and is best used when only proestrus or estrus stages need to be identified. Strain to strain differences, especially in coat color can make it difficult to determine the stage of the estrous cycle accurately by visual observation. Presented here are a series of images of the vaginal opening at each stage of the estrous cycle for 3 mouse strains of different coat colors: black (C57BL/6J), agouti (CByB6F1/J) and albino (BALB/cByJ). When all 4 stages (proestrus, estrus, metestrus, and diestrus) need to be identified, vaginal cytology is regarded as the most accurate method. An identification tool is presented to aid the user in determining the stage of estrous when using vaginal cytology. These images and descriptions are an excellent resource for learning how to determine the stage of the estrous cycle by visual observation or vaginal cytology.

Exploring the Influence of Students' Attributions for Success on Their Self-Regulation in Pathophysiology

Pathophysiology is a difficult course both for students to take and for instructors to teach. However, little research has explored learner characteristics that teachers may address through targeted instruction to make both the teaching and learning experience better. This study examined the influence of students' causal attributions for success on their self-regulated learning, which is strongly associated with positive learning outcomes. Results indicated that ability, effort, and luck attributions for success collectively influenced Pathophysiology students' self-regulated learning and that ability was the most potent influence. The findings and the implication for teaching are discussed.

Associations Between Pro- and Anti-Inflammatory Cytokine Genes and Breast Pain in Women Prior to Breast Cancer Surgery

The purposes of this study were to determine the occurrence rate for preoperative breast pain; describe the characteristics of this pain; evaluate for differences in demographic and clinical characteristics; and evaluate for variations in pro- and anti-inflammatory cytokine genes between women who did and did not report pain. Patients (n = 398) were recruited prior to surgery and completed self-report questionnaires on a number of pain characteristics. Genotyping was done using a custom genotyping array. Women (28.2%) who reported breast pain were significantly younger (P < .001); more likely to be nonwhite (P = .032); reported significantly lower Karnofsky Performance Status scores (P = .008); were less likely to be postmenopausal (P = .012); and had undergone significantly more biopsies (P = .006). Carriers of the minor allele for a single nucleotide polymorphism in interleukin (IL)1-receptor 1 (IL1R1) (rs2110726) were less likely to report breast pain prior to surgery (P = .007). Carriers of the minor allele for a single nucleotide polymorphism in IL13 (rs1295686) were more likely to report breast pain prior to surgery (P = .019). Findings suggest that breast pain occurs in over a quarter of women who are about to undergo breast cancer surgery. Based on phenotypic and genotypic characteristics found, inflammatory mechanisms contribute to preoperative breast pain. PERSPECTIVE: In women with breast cancer, preoperative pain may be associated with increases in inflammatory responses associated with an increased number of biopsies. In addition, differences in cytokine genes may contribute to this preoperative breast pain.

Measures to Prevent Transfusion-related Acute Lung Injury (TRALI)

Modest Alcohol Consumption is Associated with Decreased Prevalence of Steatohepatitis in Patients with Nonalcoholic Fatty Liver Disease (NAFLD)

BACKGROUND & AIMS: Nonalcoholic fatty liver disease (NAFLD) is a cardiovascular risk factor. Although modest alcohol consumption may reduce the risk for cardiovascular mortality, whether patients with NAFLD should be allowed modest alcohol consumption remains an important unaddressed issue. We aimed to evaluate the association between modest alcohol drinking and nonalcoholic steatohepatitis(NASH), among subjects with NAFLD. METHODS: In a Cross-sectional analysis of adult participants in the NIH NASH Clinical Research Network, only modest or non-drinkers were included: participants identified as 1) drinking > 20gm/day, 2) binge drinkers, or 3) non-drinkers with previous alcohol consumption were excluded. The odds of having a histological diagnosis of NASH and other histological features of NAFLD were analyzed using multiple ordinal logistic regression. RESULTS: The analysis included 251 lifetime non-drinkers and 331 modest drinkers. Modest drinkers compared to nondrinkers had lower odds of having a diagnosis of NASH (Summary odds ratio 0.56, 95%CI 0.39-0.84, p=0.002). The odds of NASH decreased as the frequency of alcohol consumption increased within the range of modest consumption. Modest drinkers also had significantly lower odds for fibrosis (OR 0.56 95%CI 0.41-0.77) and ballooning hepatocellular injury (OR 0.66 95%CI 0.48-0.92) than lifetime non-drinkers. CONCLUSIONS: In a large, well-characterized population with biopsy-proven NAFLD, modest alcohol consumption was associated with lesser degree of severity as determined by lower odds of the key features that comprise a diagnosis of steatohepatitis, as well as fibrosis. These findings demonstrate the need for prospective studies and a coordinated consensus on alcohol consumption recommendations in NAFLD.

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