Other articles by Elizabeth Jones on PubMed

• Hepatic Differentiation of Murine Embryonic Stem Cells Experimental Cell Research. Jan, 2002  |   Pubmed ID: 11740861 Murine embryonic stem (ES) cells can replicate indefinitely in culture and can give rise to all tissues, including the germline, when reimplanted into a murine blastocyst. ES cells can also be differentiated in vitro into a wide range of cell types. We have utilized a liver-specific marker to demonstrate that murine ES cells can differentiate into hepatocytes in vitro. We have used ES cells carrying a gene trap vector insertion (I.114) into an ankyrin repeat-containing gene (Gtar) that we have previously shown provides an exclusive beta-galactosidase marker for the early differentiation of hepatocytes in vivo. beta-Galactosidase-positive cells were differentiated from I.114 ES cells in vitro. The identity of these cells was confirmed by the expression of the proteins alpha-fetoprotein, albumin, and transferrin and by the fact that they have an ultrastructural appearance consistent with that of embryonic hepatocytes. We propose that this model system of hepatic differentiation in vitro could be used to define factors that are involved in specification of the hepatocyte lineage. In addition, human ES cells have recently been derived and it has been proposed that they may provide a source of differentiated cell types for cell replacement therapies in the treatment of a variety of diseases.
• Consumption of Fruits, Vegetables, Soft Drinks, and High-fat-containing Snacks Among Mexican Children on the Mexico-U.S. Border Archives of Medical Research. Jan-Feb, 2002  |   Pubmed ID: 11825635 The recommended diet for children would promote health, support growth, and prevent risk of disease. Diets high in fruits and vegetables demonstrate a strong and consistent pattern for decreasing the risk for many cancers and providing benefits against cardiovascular disease, diabetes, obesity, and stroke. The purpose of this study was to assess fruit, vegetable, soft drink, and high-fat-containing snack consumption among fifth- and ninth-grade children attending public schools in the northeastern Mexican state of Baja California.
• The 2001 George W. Beadle Medal. Gerald R. Fink Genetics. Feb, 2002  |   Pubmed ID: 11894820
• Annexin IV (Xanx-4) Has a Functional Role in the Formation of Pronephric Tubules Development (Cambridge, England). Apr, 2002  |   Pubmed ID: 11923205 Vertebrate kidney organogenesis is characterised by the successive formation of the pronephros, the mesonephros and the metanephros. The pronephros is the first to form and is the functional embryonic kidney of lower vertebrates; although it is vestigial in higher vertebrates, it is a necessary precursor for the other kidney types. The Xenopus pronephros is a simple paired organ; each nephron consists of a single large glomus, one set of tubules and a single duct. The simple organisation of the pronephros and the amenability of Xenopus laevis embryos to manipulation make the Xenopus pronephros an attractive system in which to study organogenesis. It has been shown that pronephric tubules can be induced to form in presumptive ectodermal tissue by treatment with RA and activin. We have used this system in a subtractive hybridisation screen that resulted in the cloning of Xenopus laevis annexin IV (Xanx-4). Xanx-4 transcripts are specifically located to the developing pronephric tubules, and the protein to the luminal surface of these tubules. Temporal expression shows zygotic transcription is upregulated at the time of pronephric tubule specification and persists throughout pronephric development. The temporal and spatial expression pattern of Xanx-4 suggests it may have a role in pronephric tubule development. Overexpression of Xanx-4 yields no apparent phenotype, but Xanx-4 depletion, using morpholinos, produces a shortened, enlarged tubule phenotype. The phenotype observed can be rescued by co-injection of Xanx-4 mRNA. Although the function of annexins is not yet clear, studies have suggested a role for annexins in a number of cellular processes. Annexin IV has been shown to have an inhibitory role in the regulation of epithelial calcium-activated chloride ion conductance. The enlarged pronephric tubule phenotype observed may be attributed to incorrect modulation of exocytosis, membrane plasticity or ion channels and/or water homeostasis. In this study, we demonstrate an in vivo role for annexin IV in the development of the pronephric tubules in Xenopus laevis.
• Lymphangioleiomyomatosis: Correlation of Qualitative and Quantitative Thin-section CT with Pulmonary Function Tests and Assessment of Dependence on Pleurodesis Radiology. Apr, 2002  |   Pubmed ID: 11930066 To explore the relationship between findings at thin-section computed tomography (CT) and pulmonary function tests in lymphangioleiomyomatosis (LAM) and to evaluate the influence of pleurodesis on this relation and the effectiveness of quantitative versus qualitative CT in the assessment of disease severity.
• The Hypertrophic Heart Rat: a New Normotensive Model of Genetic Cardiac and Cardiomyocyte Hypertrophy Physiological Genomics. 2002  |   Pubmed ID: 11948289 We describe a new line of rats with inherited cardiomyocyte and ventricular hypertrophy. From a second-generation cross of spontaneously hypertensive and Fischer 344 rats, we selected for low blood pressure and either high or low echocardiographic left ventricular (LV) mass over four generations to establish the hypertrophic heart rat (HHR) and normal heart rat (NHR) lines, respectively. After 13 generations of inbreeding, HHR had significantly greater (P < 0.0001) LV mass-to-body weight ratio (2.68 g/kg, SE 0.14) than NHR matched for age (1.94 g/kg, SE 0.02) or body weight (2.13 g/kg, SE 0.03). The isolated cardiomyocytes of HHR were significantly (P < 0.0001) longer and wider (161 microm, SE 0.83; 35.6 microm, SE 2.9) than NHR (132 microm, SE 1.2; 29.5 microm, SE 0.35). Telemetric 24-h recordings of mean arterial pressure revealed no significant differences between HHR and NHR. The HHR offers a new model of primary cardiomyocyte hypertrophy with normal blood pressure in which to examine genotypic causes and pathogenetic mechanisms of hypertrophy and its complications.
• An Enhanced Molecular Marker Based Genetic Map of Perennial Ryegrass (Lolium Perenne) Reveals Comparative Relationships with Other Poaceae Genomes Genome / National Research Council Canada = Génome / Conseil National De Recherches Canada. Apr, 2002  |   Pubmed ID: 11962626 A molecular-marker linkage map has been constructed for perennial ryegrass (Lolium perenne L.) using a one-way pseudo-testcross population based on the mating of a multiple heterozygous individual with a doubled haploid genotype. RFLP, AFLP, isoenzyme, and EST data from four collaborating laboratories within the International Lolium Genome Initiative were combined to produce an integrated genetic map containing 240 loci covering 811 cM on seven linkage groups. The map contained 124 codominant markers, of which 109 were heterologous anchor RFLP probes from wheat, barley, oat, and rice, allowing comparative relationships between perennial ryegrass and other Poaceae species to be inferred. The genetic maps of perennial ryegrass and the Triticeae cereals are highly conserved in terms of synteny and colinearity. This observation was supported by the general agreement of the syntenic relationships between perennial ryegrass, oat, and rice and those between the Triticeae and these species. A lower level of synteny and colinearity was observed between perennial ryegrass and oat compared with the Triticeae, despite the closer taxonomic affinity between these species. It is proposed that the linkage groups of perennial ryegrass be numbered in accordance with these syntenic relationships, to correspond to the homoeologous groups of the Triticeae cereals.
• Toll-like Receptor (TLR)2 and TLR4 in Human Peripheral Blood Granulocytes: a Critical Role for Monocytes in Leukocyte Lipopolysaccharide Responses Journal of Immunology (Baltimore, Md. : 1950). May, 2002  |   Pubmed ID: 11971020 Leukocyte responsiveness to LPS is dependent upon CD14 and receptors of the Toll-like receptor (TLR) family. Neutrophils respond to LPS, but conflicting data exist regarding LPS responses of eosinophils and basophils, and expression of TLRs at the protein level in these granulocyte lineages has not been fully described. We examined the expression of TLR2, TLR4, and CD14 and found that monocytes expressed relatively high levels of cell surface TLR2, TLR4, and CD14, while neutrophils also expressed all three molecules, but at low levels. In contrast, basophils expressed TLR2 and TLR4 but not CD14, while eosinophils expressed none of these proteins. Tested in a range of functional assays including L-selectin shedding, CD11b up-regulation, IL-8 mRNA generation, and cell survival, neutrophils responded to LPS, but eosinophils and basophils did not. In contrast to previous data, we found, using monocyte depletion by negative magnetic selection, that neutrophil responses to LPS were heavily dependent upon the presence of a very low level of monocytes, and neutrophil survival induced by LPS at 22 h was monocyte dependent. We conclude that LPS has little role in the regulation of peripheral blood eosinophil and basophil function, and that, even in neutrophils, monocytes orchestrate many previously observed leukocyte LPS response patterns.
• Implementation and Evaluation of a Cognitive-behavioral Intervention to Prevent Problem Behavior in a Disorganized School Prevention Science : the Official Journal of the Society for Prevention Research. Mar, 2002  |   Pubmed ID: 12002558 We assessed the effectiveness of two realizations of cognitive-behavioral instruction in an inner-city middle school with high rates of absenteeism, low staff morale, and chronic low academic achievement. Implementation measures showed that 68% of intended instruction was delivered in the first realization. Self-report measures showed improved school conduct, less victimization in school, and more positive peer associations for the treatment group than for the comparison group at the end of the school year. Treatment students were less likely to leave the school than were comparison students, but were more often absent and tardy. Implementation was poorer in the second realization, and there were no treatment-comparison differences on self-reports or teacher ratings, but treatment students less often left the school. Difficulties in conducting instruction in difficult settings may limit the effectiveness of otherwise efficacious interventions. Specific intervention programs may offer minimal benefits if more basic school improvements are not achieved.
• Vacuolar Proteases and Proteolytic Artifacts in Saccharomyces Cerevisiae Methods in Enzymology. 2002  |   Pubmed ID: 12073340
• High Prevalence of Osteonecrosis of the Femoral Head in HIV-infected Adults Annals of Internal Medicine. Jul, 2002  |   Pubmed ID: 12093241 Osteonecrosis has been reported to occur occasionally among HIV-infected patients. The diagnosis of symptomatic osteonecrosis of the hip in two of the authors' patients, together with reports from community physicians, raised a concern that the prevalence of osteonecrosis is increasing.
• Detection of Human Papillomavirus DNA in Urine Specimens from Human Immunodeficiency Virus-positive Women Journal of Clinical Microbiology. Sep, 2002  |   Pubmed ID: 12202546 Human immunodeficiency virus (HIV)-positive women may represent one of the fastest-growing populations at risk for acquiring cervical cancer and thus require frequent screening. The purpose of the present studies was to validate a PCR-based urine assay by comparing detection and genotyping of human papillomavirus (HPV) DNA in urine samples and matching cervical swab specimens of HIV-positive women. Despite a difference in amplifiability, the prevalence of any HPV genotype (58% for the cervical swab specimens and 48% for the urine specimens) was not significantly different in this population. The levels of concordance were 70, 71, and 78% for detection of any HPV type, any high-risk HPV type, or any low-risk HPV type in the two specimen types, respectively. While instances of discordant detection were greater for the cervical swab specimens than for the urine specimens, this was not statistically significant. The distributions of HPV genotypes were similar in the cervix and the urine for the majority of types examined. Importantly, detection of HPV DNA in urine was associated with an abnormal Papanicolaou smear to the same extent that detection of HPV DNA in a cervical swab specimen was. These data provide preliminary support for the proposal to use urine testing as a primary or secondary screening tool for cervical cancer in HIV-positive women or as an epidemiological tool. Additional studies with larger sample sizes must be conducted in order to further verify these findings.
• Role of the Unfolded Protein Response Pathway in Regulation of INO1 and in the Sec14 Bypass Mechanism in Saccharomyces Cerevisiae Genetics. Sep, 2002  |   Pubmed ID: 12242221 INO1, encoding inositol 1-phosphate synthase, is the most highly regulated of a class of genes containing the repeated element, UAS(INO), in their promoters. Transcription of UAS(INO)-containing genes is modulated by the availability of exogenous inositol and by signals generated by alteration of phospholipid metabolism. The unfolded protein response (UPR) pathway also is involved in INO1 expression and the ire1Delta and hac1Delta mutants are inositol auxotrophs. We examined the role of the UPR in transmitting a signal generated in response to inositol deprivation and to alteration of phospholipid biosynthesis created in the sec14(ts) cki1Delta genetic background. We report that the UPR is required for sustained high-level INO1 expression in wild-type strains, but not for transient derepression in response to inositol deprivation. Moreover, the UPR is not required for expression or regulation of INO1 in response to the change in lipid metabolism that occurs in the sec14(ts) cki1Delta genetic background. Thus, the UPR signal transduction pathway is not involved directly in transcriptional regulation of INO1 and other UAS(INO)-containing genes. However, we discovered that inactivation of Sec14p leads to activation of the UPR, and that sec14 cki1 strains exhibit defective vacuolar morphology, suggesting that the mechanism by which the cki1Delta mutation suppresses the growth and secretory defect of sec14 does not fully restore wild-type morphology. Finally, synthetic lethality involving sec14 and UPR mutations suggests that the UPR plays an essential role in survival of sec14 cki1 strains.
• Colorectal Cancer Screening with Double-contrast Barium Enema: a National Survey of Diagnostic Radiologists AJR. American Journal of Roentgenology. Dec, 2002  |   Pubmed ID: 12438029 This article describes diagnostic radiologists' colorectal cancer screening activities and beliefs about screening effectiveness and future capacity for screening with double-contrast barium enema, and compares radiologists' opinions about colorectal cancer screening with those of primary care physicians.
• Functional Electrical Stimulation for a Dropped Foot Journal of Long-term Effects of Medical Implants. 2002  |   Pubmed ID: 12545941 The purpose of this collective review is to study the techniques, usage, methods, and clinical results of functional electrical stimulation applied to the peroneal nerve in the treatment of dropped foot. When stimulation is applied through surface electrodes, clinical reports have documented the therapeutic and orthotic benefits of functional electrical stimulation. Clinical trials are now being undertaken in which implantable electrodes are being used to stimulate the peroneal nerve in patients with dropped foot.
• Functional Electrical Stimulation Cycle Ergometer Exercise for Spinal Cord Injured Patients Journal of Long-term Effects of Medical Implants. 2002  |   Pubmed ID: 12545942 The purpose of this collective review on functional electrical stimulation (FES) cycle ergometer training is to describe the pathologic effects of spinal cord injury (SCI) and the structure and function of the FES cycle ergometers, which reverse the devastating systemic and life-threatening effects of SCI. The pathophysiologic consequences of SCI include diminished cardiopulmonary and circulatory function as well as lower extremity muscle atrophy and bone mass reduction. Clinical studies have demonstrated that the two FES cycle ergometers offer promise in reversing these devastating consequences of SCI, which can shorten patients' lives. On the basis of this collective review, it is recommended that all patients with SCI have the benefits of this potentially life-sustaining clinical modality.
• Functional Electrical Stimulation in Tetraplegic Patients to Restore Hand Function Journal of Long-term Effects of Medical Implants. 2002  |   Pubmed ID: 12545943 The purpose of this collective review is to describe a new form of functional electrical stimulation called neuroprosthesis. This unique technology has been devised to produce lateral pinch and palmar grasp in persons with C5 and C6 motor level spinal cord injuries. This neuroprosthesis includes external as well as implanted components. First, a receiver is surgically implanted into the patient's chest above a pectoralis major muscle. The receiver stimulator is then connected to 8 surgically implanted epimysial or intramuscular electrodes. Restoration of upper extremity function can greatly improve the lives of people affected with tetraplegia. When contralateral shoulder movements trigger an external transmitting coil, it sends a radio wave impulse to the stimulator inducing contraction of the muscles. Many tetraplegics are regaining hand function using implanted functional electrical stimulation. One major limitation is that the key muscles to be stimulated may have lower motor neuron damage, but this obstacle has been successfully overcome using surgical modifications of the biomechanics of the hand.
• Functional Electrical Stimulation of Bladder and Bowel in Spinal Cord Injury Journal of Long-term Effects of Medical Implants. 2002  |   Pubmed ID: 12545944 The purpose of this collective review is to examine the use of functional electrical stimulation for incontinence. The Finetech-Brindley bladder system enhances voiding through stimulation via electrodes implanted around the ventral sacral roots. Detrusor hyperreflexia is eliminated through complete dorsal rhizotomy, which results in loss of reflex defecation and reflex erection/reflex lubrication. Consequently, a new system is being devised in which functional electrical stimulation for incontinence in spinal cord injury can be achieved without dorsal rhizotomy.
• Functional Electrical Stimulation for Ejaculation Journal of Long-term Effects of Medical Implants. 2002  |   Pubmed ID: 12545945 The purpose of this collective review is to discuss the technique and effectiveness of rectal probe electroejaculation in conjunction with various assistive reproductive modalities. The electroejaculation probe is positioned inside the anal ring with the electrodes placed against the prostate gland and seminal vesicles, after which electrostimulation is begun. Rectal probe electroejaculation must be always employed in a hospital setting to detect and prevent autonomic dysreflexia. This technology is considered the best approach in patients with spinal cord injury levels below T10 or in other patients in which penile vibratory stimulation fails.
• Diagnosis of Avascular Necrosis of the Hip in Asymptomatic HIV-infected Patients: Clinical Correlation of Physical Examination with Magnetic Resonance Imaging Journal of Back and Musculoskeletal Rehabilitation. Jan, 2002  |   Pubmed ID: 22387437 Objective: To determine if physical examination can identify avascular necrosis of the hip (AVN) in asymptomatic HIV-infected patients.Design: Prospective, blinded population studyResults: Ten of the 176 patients were positive for AVN by MRI. Four subjects had unilateral disease and six had bilateral disease. Five hips (1.4%) in four patients were indeterminate. We evaluated physical examination maneuvers both singly and in combination. Tests done singly generally provided a higher degree of specificity (67-92%) but sensitivities were lower (0-50%) with all p-values ≥0.08. Positive predictive values based on physical exam, were 90% for any single test. Combining all tests gave a high sensitivity (88%) and negative predictive value (98%), but low specificity (34%) and positive predictive value (6%) with p = 0.10. Only two of 16 hips with positive MRI findings showed no abnormalities when all tests were combinedConclusions: This study establishes the limited usefulness of a detailed physical examination of the hip early in the course of AVN. Patients who test negative on physical exam are unlikely to have AVN positive by MRI. Positive findings on physical examination of the hip may help identify patients who need further evaluation by MRI based on overall clinical suspicion.
• A National Survey of Primary Care Physicians' Colorectal Cancer Screening Recommendations and Practices Preventive Medicine. Mar, 2003  |   Pubmed ID: 12634026 National data on providers' colorectal cancer (CRC) screening knowledge, attitudes, and practices are sparse. This study assessed primary care physicians' (PCPs') beliefs about the effectiveness of CRC screening, their recommendations for screening, their perceptions of the influence of published guidelines on their CRC screening recommendations, and how they conduct CRC screening in their clinical practices.
• NFAT4 is Expressed in Primary Astrocytes and Activated by Glutamate Journal of Neuroscience Research. Apr, 2003  |   Pubmed ID: 12671993 Calcium signaling is a critical component of astrocyte activation but little information is available regarding the identity and function of transcriptional targets of calcium signaling in these cells. As a first step in elucidating the mechanisms that astrocytes use to regulate transcription in response to raised intracellular calcium concentrations, we have investigated expression and activation of the calcium activated transcription factors of the NFAT family. We report here that NFAT4 is expressed in U373 astrocytoma cells and in primary cultures of astrocytes. Treatment of U373 cells or primary astrocytes with the calcium ionophore A23187, or the neurotransmitter glutamate, leads to NFAT nuclear translocation and increased DNA binding to a NFAT consensus site through a cyclosporin A-sensitive process. These data suggest that NFAT4 functions as a calcium-responsive transcription factor in astrocytes.
• Selective Roles for Toll-like Receptor (TLR)2 and TLR4 in the Regulation of Neutrophil Activation and Life Span Journal of Immunology (Baltimore, Md. : 1950). May, 2003  |   Pubmed ID: 12734376 Neutrophil responses to commercial LPS, a dual Toll-like receptor (TLR)2 and TLR4 activator, are regulated by TLR expression, but are amplified by contaminating monocytes in routine cell preparations. Therefore, we investigated the individual roles of TLR2 and TLR4 in highly purified, monocyte-depleted neutrophil preparations, using selective ligands (TLR2, Pam(3)CysSerLys(4) and Staphylococcus aureus peptidoglycan; TLR4, purified LPS). Activation of either TLR2 or TLR4 caused changes in adhesion molecule expression, respiratory burst (alone, and synergistically with fMLP), and IL-8 generation, which was, in part, dependent upon p38 mitogen-activated protein kinase signaling. Neutrophils also responded to Pam(3)CysSerLys(4) and purified LPS with down-regulation of the chemokine receptor CXCR2 and, to a lesser extent, down-regulation of CXCR1. TLR4 was the principal regulator of neutrophil survival, and TLR2 signals showed relatively less efficacy in preventing constitutive apoptosis over short time courses. TLR4-mediated neutrophil survival depended upon signaling via NF-kappa B and mitogen-activated protein kinase cascades. Prolonged neutrophil survival required both TLR4 activation and the presence of monocytes. TLR4 activation of monocytes was associated with the release of neutrophil survival factors, which was not evident with TLR2 activation, and TLR2 activation in monocyte/neutrophil cocultures did not prevent late neutrophil apoptosis. Thus, TLRs are important regulators of neutrophil activation and survival, with distinct and separate roles for TLR2 and TLR4 in neutrophil responses. TLR4 signaling presents itself as a pharmacological target that may allow therapeutic modulation of neutrophil survival by direct and indirect mechanisms at sites of inflammation.
• Isolation and Growth Factor Inducibility of the Xenopus Laevis Lmx1b Gene The International Journal of Developmental Biology. May, 2003  |   Pubmed ID: 12755330 This paper reports the cloning of the full length Xenopus laevis Lmx1b gene, Xlmx1b. Xlmx1b is a LIM homeodomain protein with high conservation to homologues identified in human, mouse, hamster and chick. In situ hybridisation and RT-PCR analysis showed that Xlmx1b has a specific temporal expression pattern which can be separated into three main spatial domains. An Xlmx1b probe hybridized to regions of the nervous system from stage 13 onwards; these regions included the placodes and otic vesicles, the eye and specific sets of neurons. Sectioning of in situ hybridised embryos confirmed the location of transcripts as discreet regions of staining in ventrolateral regions of the neural tube. From stage 27, transcripts could be detected in the capsule of pronephric glomus. Finally, transcripts were detected by Northern blot analysis in the developing fore and hind limbs. Xlmx1b transcripts were also detected by Northern blot analysis in eye, brain, muscle and mesonephros tissue in metamorphosing tadpoles. RT-PCR analysis showed that zygotic expression of Xlmx1b is initiated at stage 10.5 and the temporal sequence of Xlmx1b expression is identical in both neural and presumptive pronephros regions. The effects of the growth factors activin A, retinoic acid (RA) and basic fibroblast growth factor (bFGF) on the regulation of Xlmx1b were also studied. Xlmx1b was found to be upregulated by activin A and RA inhibited this upregulation in a concentration dependant manner. In contrast, bFGF had no effect on the regulation of Xlmx1b.
• Aortic Source of Brain Embolism Current Treatment Options in Cardiovascular Medicine. Jul, 2003  |   Pubmed ID: 12777199 Aortic arch atheroma has more recently been identified as an independent risk factor for ischemic stroke. Initially, this was a result of careful autopsy observations, then followed by a series of in vivo studies in which aortic arch atheroma was identified by transesophageal echocardiography. The association of aortic arch atheroma with ischemic stroke is most likely causal, given that the stroke risk increases with increasing thickness of arch atheroma. There is quite a sharp increase in stroke risk for atheroma of 4 mm or greater compared with lesser thicknesses. The clinical diagnosis is suggested when transient ischemic attack or ischemic stroke has occurred in which no obvious cardiac or arterial source of embolism is found. The presence of aortic arch atheroma is usually detected by transesophageal echocardiography and sometimes by magnetic resonance imaging or computed tomography. There is uncertainty about clinical management, particularly for secondary prevention. Options include the use of antiplatelet agents, anticoagulants, thrombolysis, or surgery. The latter two options have only been described rarely in case reports. Of the less invasive approaches, combination antiplatelet therapy with aspirin and clopidogrel is favored, or the use of warfarin. The Aortic arch Related Cerebral Hazard (ARCH) trial is being conducted to determine which of these is more effective in minimizing a composite outcome cluster of ischemic stroke, intracranial hemorrhage, myocardial infarction, peripheral embolism, or vascular death. Other more general management strategies should include reasonably aggressive risk factor control with blood pressure and lipid-lowering therapies and, if indicated, careful diabetic control.
• Isolation and Characterisation of an Invertase CDNA from Perennial Ryegrass (Lolium Perenne) Journal of Plant Physiology. Aug, 2003  |   Pubmed ID: 12964866 An invertase (LpFT2) cDNA from perennial ryegrass was isolated and sequenced. Nucleotide sequence analysis revealed an ORF of 2016 bp encoding a protein of 671 amino acids. LpFT2 is 76% identical to sugarcane soluble acid invertase, and contains invertase and fructosyltransferase functional domains. LpFT2 is present as a single copy gene and maps to the distal region of LG6 in perennial ryegrass. The expression pattern analysis of LpFT2 revealed transcript accumulation in seedlings and in mature leaf sheaths. The LpFT2 recombinant protein expressed in yeast showed invertase and fructan exohydrolase-like activities with complete breakdown of sucrose, 1-kestose (DP3), 1,1-kestotetraose (DP4) and 1,1,1-kestopentaose (DP5) into glucose and fructose.
• Assessing Probability of Ancestry Using Simple Sequence Repeat Profiles: Applications to Maize Inbred Lines and Soybean Varieties Genetics. Sep, 2003  |   Pubmed ID: 14504240 Determining parentage is a fundamental problem in biology and in applications such as identifying pedigrees. Difficulties inferring parentage derive from extensive inbreeding within the population, whether natural or planned; using an insufficient number of hypervariable loci; and from allele mismatches caused by mutation or by laboratory errors that generate false exclusions. Many studies of parentage have been limited to comparisons of small numbers of specific parent-progeny triplets. There have been few large-scale surveys of candidates in which there is no prior knowledge of parentage. We present an algorithm that determines the probability of parentage in circumstances where there is no prior knowledge of pedigree and that is robust in the face of missing data and mistyped data. The focus is parentage of an inbred line having uncertain ancestry. The algorithm is a variation of a previously published hybrid-focused algorithm. We describe the algorithm and demonstrate its performance in determining parentage of 43 inbred varieties of soybean that have been profiled using 236 SSR loci and from seven inbred varieties of maize that were profiled using 70 SSR loci. We include simulations of additional levels of missing and mistyped data to show the algorithm's utility and flexibility.
• Increases in CD4+ T Lymphocytes Occur Without Increases in Thymic Size in HIV-infected Subjects Receiving Interleukin-2 Therapy Journal of Acquired Immune Deficiency Syndromes (1999). Nov, 2003  |   Pubmed ID: 14600575 To examine the potential contribution of the thymus to CD4+ T-lymphocyte increases in HIV-infected patients receiving intermittent interleukin-2 (IL-2) therapy.
• Management of an Ileostomy and Mucous Fistula Located in a Dehisced Wound in a Patient with Morbid Obesity Journal of Wound, Ostomy, and Continence Nursing : Official Publication of The Wound, Ostomy and Continence Nurses Society / WOCN. Nov, 2003  |   Pubmed ID: 14615767
• Optimization of PCR Based Detection of Human Papillomavirus DNA from Urine Specimens Journal of Clinical Virology : the Official Publication of the Pan American Society for Clinical Virology. Apr, 2004  |   Pubmed ID: 15018850 Human papillomavirus (HPV) causes cervical cancer. Current screening requires a yearly pelvic exam and Pap smear. However, these procedures are impractical for screening all women at risk for disease. Urine sampling has been successfully utilized to screen for Chlamydia trachomatis (CT) and Neisseria gonorrhoreae (NG) infections and has been considered for HPV DNA detection by several investigators. However, no study to date has been performed to specifically optimize HPV detection in urine.
• The Sec1/Munc18 Protein, Vps33p, Functions at the Endosome and the Vacuole of Saccharomyces Cerevisiae Molecular Biology of the Cell. Jun, 2004  |   Pubmed ID: 15047864 The Sec1/Munc18 (SM) family of proteins is thought to impart compartmental specificity to vesicle fusion reactions. Here we report characterization of Vps33p, an SM family member previously thought to act exclusively at the vacuolar membrane with the vacuolar syntaxin Vam3p. Vacuolar morphology of vps33Delta cells resembles that of cells lacking both Vam3p and the endosomal syntaxin Pep12p, suggesting that Vps33p may function with these syntaxins at the vacuole and the endosome. Consistent with this, vps33 mutants secrete the Golgi precursor form of the vacuolar hydrolase CPY into the medium. We also demonstrate that Vps33p acts at other steps, for vps33 mutants show severe defects in endocytosis at the late endosome. At the endosome, Vps33p and other class C members exist as a complex with Vps8p, a protein previously known to act in transport between the late Golgi and the endosome. Vps33p also interacts with Pep12p, a known interactor of the SM protein Vps45p. High copy PEP7/VAC1 suppresses vacuolar morphology defects of vps33 mutants. These findings demonstrate that Vps33p functions at multiple trafficking steps and is not limited to action at the vacuolar membrane. This is the first report demonstrating the involvement of a single syntaxin with two SM proteins at the same organelle.
• QTL Analysis of Quantitative Resistance to Phytophthora Infestans (late Blight) in Tomato and Comparisons with Potato Genome / National Research Council Canada = Génome / Conseil National De Recherches Canada. Jun, 2004  |   Pubmed ID: 15190365 Quantitative trait loci (QTLs) for resistance to Phytophthora infestans (late blight) were mapped in tomato. Reciprocal backcross populations derived from cultivated Lycopersicon esculentum x wild Lycopersicon hirsutum (BC-E, backcross to L. esculentum; BC-H, backcross to L. hirsutum) were phenotyped in three types of replicated disease assays (detached-leaflet, whole-plant, and field). Linkage maps were constructed for each BC population with RFLPs. Resistance QTLs were identified on all 12 tomato chromosomes using composite interval mapping. Six QTLs in BC-E (lb1a, lb2a, lb3, lb4, lb5b, and lb11b) and two QTLs in BC-H (lb5ab and lb6ab) were most consistently detected in replicated experiments or across assay methods. Lycopersicon hirsutum alleles conferred resistance at all QTLs except lb2a. Resistance QTLs coincided with QTLs for inoculum droplet dispersal on leaves, a trait in L. hirsutum that may contribute to resistance, and dispersal was mainly associated with leaf resistance. Some P. infestans resistance QTLs detected in tomato coincided with chromosomal locations of previously mapped R genes and QTLs for resistance to P. infestans in potato, suggesting functional conservation of resistance within the Solanaceae.
• Randomized Phase II Trial of Docetaxel Plus Thalidomide in Androgen-independent Prostate Cancer Journal of Clinical Oncology : Official Journal of the American Society of Clinical Oncology. Jul, 2004  |   Pubmed ID: 15226321 Both docetaxel and thalidomide have demonstrated activity in androgen-independent prostate cancer (AIPC). We compared the efficacy of docetaxel to docetaxel plus thalidomide in patients with AIPC.
• The Role of Interleukin-1beta in Direct and Toll-like Receptor 4-mediated Neutrophil Activation and Survival The American Journal of Pathology. Nov, 2004  |   Pubmed ID: 15509550 The regulation of systemic and local neutrophil activation is crucial to the clearance of infections and the successful resolution of inflammation without progress to tissue damage or disseminated inflammatory reactions. Using purified lipopolysaccharide (pLPS) and highly purified neutrophils, we have previously shown that Toll-like receptor 4 signaling is a potent neutrophil activator, but a poor stimulator of survival. In the presence of peripheral blood mononuclear cells (PBMCs), however, pLPS becomes a potent neutrophil survival factor. Interleukin (IL)-1beta has been identified as an important neutrophil activator and prosurvival cytokine, and is produced in abundance by LPS-stimulated PBMCs. We now show that IL-1beta fails to activate highly purified neutrophils or enhance their survival, but in the presence of PBMCs, IL-1beta induces neutrophil survival. We hypothesized that LPS-primed neutrophils might become responsive to IL-1beta, but were unable to demonstrate this. Moreover, IL-1ra failed to prevent pLPS + PBMC-dependent neutrophil survival. In studies of IL-1R1(-/-) mice, we found that LPS was still able to mediate neutrophil survival, and neutrophil survival was enhanced by the addition of monocytic cells. Thus an important paradigm of neutrophil regulation needs to be viewed in the context of a cellular network in which actions of IL-1beta on neutrophils are indirect and mediated by other cells.
• Cloning and Characterisation of the Immunophilin X-CypA in Xenopus Laevis Gene Expression Patterns : GEP. Nov, 2004  |   Pubmed ID: 15533818 This paper reports the cloning of Xenopus laevis, cyclophilin A gene, X-CypA. This study is the first developmental and functional characterisation in vivo of cyclophilin A in a vertebrate. X-CypA belongs to the superfamily of the immunophilin/PPIase proteins that can bind the immunosuppressant drug Cyclosporin A. Sequence analysis showed that X-CypA is highly conserved during evolution. RT-PCR and in situ hybridisation analysis showed that X-CypA expression is regulated during development and its transcripts are found in three major expression domains: nervous system, sensory organs and pronephros. Over-expression of X-CypA in embryos, analysed by in situ hybridisation and RT-PCR, leads to an expansion and disorganisation of the neural crest domain.
• Evaluating Preterm Breastfeeding Training The Practising Midwife. Oct, 2004  |   Pubmed ID: 15536817
• X-epilectin: a Novel Epidermal Fucolectin Regulated by BMP Signalling The International Journal of Developmental Biology. Dec, 2004  |   Pubmed ID: 15602698 This paper reports the cloning and characterisation of a new posterior epidermal marker, X-epilectin, in Xenopus laevis. This gene encodes for a fucolectin, which belongs to the lectin superfamily of carbohydrate binding proteins and specifically binds fucose residues. RT-PCR and in situ hybridisation show that the expression of this gene is switched on during gastrulation and up-regulated during neurula stages and found expressed ubiquitously throughout the epidermis. From tailbud stages, the expression is limited to the dorsal posterior region of the embryo, suggesting that X-epilectin expression is regulated along anteroposterior and dorsoventral gradients during development. In the adult, X-epilectin is mainly expressed in intestinal components, kidney, spinal cord and skin. The effects of growth factors on the regulation of X-epilectin were studied. Change of the fate of animal caps into cement gland or dorsal mesoderm induces a down-regulation of X-epilectin expression in explants treated respectively with ammonium chloride and activin A. We also show that X-epilectin expression is down-regulated by Noggin and tBR and that this effect is inhibited by BMP4 over-expression, suggesting X-epilectin expression is mediated by the BMP signalling pathway.
• Breastfeeding and Returning to Work The Practising Midwife. Dec, 2004  |   Pubmed ID: 15624531
• Dynamic in Vivo Imaging of Mammalian Hematovascular Development Using Whole Embryo Culture Methods in Molecular Medicine. 2005  |   Pubmed ID: 15492409 The yolk sac is the initial site of hematopoiesis in the mammalian embryo. As the embryo develops, blood vessels form around primitive erythroblasts to connect the yolk sac to the embryo, delivering newly formed blood cells to the embryonic circulation. The limited accessibility of the mammalian embryo has made it difficult to study the dynamic changes in cellular development during the formation of the early hematovascular system. Therefore, we have developed a culture system for studying early hematopoiesis, vasculogenesis. and angiogenesis in the mouse embryo. Early embryos (E7.5-E9.5) can be grown on the microscope stage to study the dynamics as vessels form and circulation begins. In addition, this mouse embryo culture system provides an excellent model for understanding the interplay between flow dynamics and cellular development.
• Xenopus: a Prince Among Models for Pronephric Kidney Development Journal of the American Society of Nephrology : JASN. Feb, 2005  |   Pubmed ID: 15647339 Recent advances in techniques that are available to study the molecular development of the frog Xenopus make developmental studies using this amphibian amenable to experimentation. This review outlines some of the attractive features of this model organism and describes how these techniques can be and are being used in studies on the organogenesis of the larval amphibian kidney, the pronephros. The roles of micromanipulation, grafting, and in vitro culturing of animal caps are discussed as tools in the analysis of kidney development and as a source of tissue for subtractive hybridization strategies. The importance of expression cloning and functional analysis of newly identified pronephros-specific genes are also described. Finally, transgenesis and electroporation are discussed as potentially new methods of gene delivery to the pronephros. These techniques can be used to help identify the gene networks that control organogenesis of this larval kidney form, which will undoubtedly have applicability to higher vertebrate kidney development.
• A Randomized, Phase II Trial of Ketoconazole Plus Alendronate Versus Ketoconazole Alone in Patients with Androgen Independent Prostate Cancer and Bone Metastases The Journal of Urology. Mar, 2005  |   Pubmed ID: 15711271 Alendronate (AL), a potent oral bisphosphonate, blocks the secretion of matrix metalloproteinase-2 and the establishment of bone metastases in animal models. Ketoconazole (KT) has demonstrated activity in androgen independent prostate cancer (AIPC). In this study we determined whether KT plus AL produced acceptable disease responses compared with KT alone. As the experimental design, 72 patients with progressive AIPC metastatic to bone were randomized to receive KT (1,200 mg daily) plus hydrocortisone (H) (30 mg daily) with or without AL (40 mg daily). Prostate specific antigen (PSA) consensus criteria and radiographic scans were used to determine the proportion of patients with a PSA decrease, time to progression and response duration. The pharmacokinetics of KT and AL were characterized and changes in circulating angiogenic factors were assessed.
• Developmental Expression of Pod 1 in Xenopus Laevis The International Journal of Developmental Biology. 2005  |   Pubmed ID: 15744669 The basic helix-loop-helix transcription factor, Pod 1, has been shown to be expressed in the mesenchyme of many developing mouse organs, including the heart, lungs and gut. In the kidneys of developing mice, Pod 1 is highly expressed in the condensing metanephric mesenchyme, differentiating and late stromal cells and in developing podocytes. We have obtained an EST (CF270487) which contains the Xenopus laevis Pod 1 sequence. Conceptual translation of the Xenopus laevis Pod 1 sequence shows approximately 85% similarity to other vertebrate homologues. RT-PCR indicates that expression is initiated at stage 13 and increases differentially in the developing pronephros compared to the whole embryo. RT-PCR of a kidney dissection at stage 42 shows higher expression in the glomus than in the tubule or duct. In situ hybridisation analysis at tail bud stages shows the anterior-most branchial arch and pronephric glomus are intensely stained. At stage 40, staining persists in the glomus and in the epicardium region of the heart. Adult organ analysis shows expression is highest in the rectum and the spleen, with significant expression in the duodenum, heart, kidney, lungs, pancreas, skin, liver and muscle.
• Age-dependent Incidence, Time Course, and Consequences of Thymic Renewal in Adults The Journal of Clinical Investigation. Apr, 2005  |   Pubmed ID: 15776111 Homeostatic regulation of T cells involves an ongoing balance of new T cell generation, peripheral expansion, and turnover. The recovery of T cells when this balance is disrupted provides insight into the mechanisms that govern homeostasis. In a long-term, single cohort study, we assessed the role of thymic function after autologous transplant in adults, correlating serial computed tomography imaging of thymic size with concurrent measurements of peripheral CD4(+) T cell populations. We established the age-dependent incidence, time course, and duration of thymic enlargement in adults and demonstrated that these changes were correlated with peripheral recovery of naive CD45RA(+)CD62L(+) and signal-joint TCR rearrangement excision circle-bearing CD4(+) populations with broad TCR diversity. Furthermore, we demonstrated that renewed thymopoiesis was critical for the restoration of peripheral CD4(+) T cell populations. This recovery encompassed the recovery of normal CD4(+) T cell numbers, a low ratio of effector to central memory cells, and a broad repertoire of TCR Vbeta diversity among these memory cells. These data define the timeline and consequences of renewal of adult thymopoietic activity at levels able to quantitatively restore peripheral T cell populations. They further suggest that structural thymic regrowth serves as a basis for the regeneration of peripheral T cell populations.
• Regulation of Human Neutrophil Chemokine Receptor Expression and Function by Activation of Toll-like Receptors 2 and 4 Immunology. May, 2005  |   Pubmed ID: 15819701 Neutrophil chemokine receptor expression can be altered by exposure to Toll-like receptor (TLR) agonists, a process that is thought to have the potential to localize neutrophils to sites of infection. In order to investigate this process in more detail, we examined the regulation of highly pure neutrophil CXCR1 and CXCR2 expression and function by selective agonists of TLR2 (Pam(3)CSK(4)) and TLR4 (lipopolysaccharide, LPS). CXCR1 and CXCR2 were down-regulated by TLR engagement. CXCR2 loss was more rapid and showed a dependence upon soluble helper molecules (LPS binding protein and CD14) that was not evident for CXCR1, suggesting differential coupling of LPS signalling to CXCR1 and CXCR2 loss. However, TLR engagement in highly pure neutrophils did not result in complete loss of chemokine receptors, and LPS-treated neutrophils remained able to mount a respiratory burst to CXCL8 and CXCL1, and were able to migrate towards CXCL8 in assays of under-agarose chemotaxis. Thus, although treatment of purified human neutrophils with TLR2 and TLR4 agonists modifies chemokine receptor expression, remaining receptors remain functionally competent.
• 'Mature' Nerve Growth Factor is a Minor Species in Most Peripheral Tissues Neuroscience Letters. May 20-27, 2005  |   Pubmed ID: 15854765 The classic neurotrophin hypothesis is based on the idea that innervating neurons derive 'mature' neurotrophin provided by the target for their survival. Yet large precursor forms of the neurotrophin nerve growth factor (NGF) have been reported in both central and peripheral tissues. In the present study, immunoblotting was used to survey peripheral tissues containing NGF-responsive neurons and to characterize various NGF species. These results demonstrate that 'mature' forms of NGF, i.e., the 13 and 16kDa species, are rare in sympathetic and sensory ganglia and in their peripheral targets, and that large molecular weight NGF precursors are abundant. In addition, certain NGF forms predominate in a given tissue, with each tissue exhibiting a characteristic NGF expression pattern. These findings suggest that NGF processing in peripheral tissues and in NGF-responsive ganglia may involve a variety of NGF species.
• Using a Histone Yellow Fluorescent Protein Fusion for Tagging and Tracking Endothelial Cells in ES Cells and Mice Genesis (New York, N.Y. : 2000). Jul, 2005  |   Pubmed ID: 15986455 We report the first endothelial lineage-specific transgenic mouse allowing live imaging at subcellular resolution. We generated an H2B-EYFP fusion protein which can be used for fluorescent labeling of nucleosomes and used it to specifically label endothelial cells in mice and in differentiating embryonic stem (ES) cells. A fusion cDNA encoding a human histone H2B tagged at its C-terminus with enhanced yellow fluorescent protein (EYFP) was expressed under the control of an Flk1 promoter and intronic enhancer. The Flk1::H2B-EYFP transgenic mice are viable and high levels of chromatin-localized reporter expression are maintained in endothelial cells of developing embryos and in adult animals upon breeding. The onset of fluorescence in differentiating ES cells and in embryos corresponds with the beginning of endothelial cell specification. These transgenic lines permit real-time imaging in normal and pathological vasculogenesis and angiogenesis to track individual cells and mitotic events at a level of detail that is unprecedented in the mouse.
• Phase III Randomized Trial of Patient-specific Vaccination for Previously Untreated Patients with Follicular Lymphoma in First Complete Remission: Protocol Summary and Interim Report Clinical Lymphoma. Jun, 2005  |   Pubmed ID: 15989711
• A Phase II Study of Perifosine in Androgen Independent Prostate Cancer Cancer Biology & Therapy. Oct, 2005  |   Pubmed ID: 16138006 Perifosine is an alkylphospholipid that has exhibited broad antineoplastic activity in preclinical studies. The primary objective of this study was to determine the clinical efficacy of this agent in the treatment of androgen-independent prostate cancer (AIPC) using PSA and clinical criteria.
• Platypnea-orthodeoxia Associated with a Fenestrated Atrial Septal Aneurysm: Case Report Cardiovascular Ultrasound. 2005  |   Pubmed ID: 16159401 Platypnea-orthodeoxia describes the condition of combined dyspnea and hypoxia respectively, whilst in the upright position, which improves in the recumbent position.
• Endotoxin Tolerance Induces Selective Alterations in Neutrophil Function Journal of Leukocyte Biology. Dec, 2005  |   Pubmed ID: 16244113 Endotoxin tolerance has the potential to limit phagocyte responses to Toll-like receptor (TLR) agonists, but the role of tolerance in regulating neutrophil responses is unknown. We investigated neutrophil responses to prolonged lipopolysaccharide (LPS) exposure and observed induction of tolerance in intracellular signaling pathways and respiratory burst. These effects were not prevented by granulocyte macrophage-colony stimulating factor (GM-CSF) pretreatment, and tolerized neutrophils retained the ability to respond to GM-CSF and other survival factors with a delay in apoptosis. In addition, LPS-exposed neutrophils showed continued generation of CXC chemokine ligand 8, which was not reduced in tolerized cells. Induction of tolerance was associated with a loss of TLR4 surface expression. Tolerance, therefore, induces a selective reprogramming of neutrophil function, but cells retain a predominantly proinflammatory phenotype.
• The Human DEK Proto-oncogene is a Senescence Inhibitor and an Upregulated Target of High-risk Human Papillomavirus E7 Journal of Virology. Nov, 2005  |   Pubmed ID: 16254365 The human DEK proto-oncogene is a nucleic acid binding protein with suspected roles in human carcinogenesis, autoimmune disease, and viral infection. Intracellular DEK functions, however, are poorly understood. In papillomavirus-positive cervical cancer cells, downregulation of viral E6/E7 oncogene expression results in cellular senescence. We report here the specific repression of DEK message and protein levels in senescing human papillomavirus type 16- (HPV16-) and HPV18-positive cancer cell lines as well as in primary cells undergoing replicative senescence. Cervical cancer cell senescence was partially overcome by DEK overexpression, and DEK overexpression was sufficient for extending the life span of primary keratinocytes, supporting critical roles for this molecule as a senescence regulator. In order to determine whether DEK is a bona fide HPV oncogene target in primary cells, DEK expression was monitored in human keratinocytes transduced with HPV E6 and/or E7. The results identify high-risk HPV E7 as a positive DEK regulator, an activity that is not shared by low-risk HPV E7 protein. Experiments in mouse embryo fibroblasts recapitulated the observed E7-mediated DEK induction and demonstrated that both basal and E7-induced regulation of DEK expression are controlled by the retinoblastoma protein family. Taken together, our results suggest that DEK upregulation may be a common event in human carcinogenesis and may reflect its senescence inhibitory function.
• Automated External Defibrillator Deployment in High Schools and Senior Centers Prehospital Emergency Care : Official Journal of the National Association of EMS Physicians and the National Association of State EMS Directors. Oct-Dec, 2005  |   Pubmed ID: 16263669 Policymakers with limited funds have been forced to make difficult decisions regarding which sites merit automated external defibrillators (AEDs). Guidelines have recommended that the allocation of AEDs be based largely on the site-specific risk of sudden cardiac death (cardiac arrest), with devices preferentially located at high-risk venues. However, there are limited data on whether such a strategy is being followed. The authors surveyed low-risk (schools) and high-risk (senior centers) venues to assess the availability of AEDs.
• Distal Enhancer Elements Transcribe Intergenic RNA in the IL-10 Family Gene Cluster Journal of Immunology (Baltimore, Md. : 1950). Dec, 2005  |   Pubmed ID: 16301651 The IL-10 gene and homologs IL-19, IL-20, and IL-24 are expressed within a highly conserved 145-kb cytokine gene cluster. Like the Th2 IL-4 cytokine gene cluster, it is feasible that there is coordinate regulation of these cytokines by distal regulatory elements spanning the locus. We initiated a search to characterize regulatory elements within the IL-10 family locus and present data herein on a conserved 40-kb region between the IL-19 and IL-10 genes. We map the location of 17 DNase I-hypersensitive sites in different murine T cell populations and identify three enhancer elements, which function in T cells in vitro. Two of these enhancer elements, located 9 kb upstream and 6.45 kb downstream of IL-10, display cell-specific function in the Th1-Th2 cell clones AE7 and D10 and also exhibit basic promoter activity. The downstream element, IL-10CNS+6.45, binds AP-1 in the absence of NFAT and expresses intergenic RNA in a Th2-specific manner, further validating its role as a Th2-specific enhancer/promoter element. We show that the five most highly conserved noncoding sequences in the 40-kb region transcribe intergenic RNA; four of these regions possess promoter activity in vitro that could account for the expression of these transcripts. Hence, we speculate that these novel regulatory elements in the IL-10 family gene locus function via an intermediate regulatory RNA.
• Abruptio Placentae Associated with Misoprostol Use in Women with Preeclampsia The Journal of Reproductive Medicine. Sep, 2005  |   Pubmed ID: 16363752 To evaluate complications associated with cervical ripening with vaginal administration of misoprostol and dinoprostone vaginal inserts in women with preeclampsia. preeclampsia.
• Analysis of Fcgamma Receptor Haplotypes in Rheumatoid Arthritis: FCGR3A Remains a Major Susceptibility Gene at This Locus, with an Additional Contribution from FCGR3B Arthritis Research & Therapy. 2006  |   Pubmed ID: 16356189 The Fcgamma receptors play important roles in the initiation and regulation of many immunological and inflammatory processes, and genetic variants (FCGR) have been associated with numerous autoimmune and infectious diseases. The data in rheumatoid arthritis (RA) are conflicting and we previously demonstrated an association between FCGR3A and RA. In view of the close molecular proximity with FCGR2A, FCGR2B and FCGR3B, additional polymorphisms within these genes and FCGR haplotypes were examined to refine the extent of association with RA. Biallelic polymorphisms in FCGR2A, FCGR2B and FCGR3B were examined for association with RA in two well characterized UK Caucasian and North Indian/Pakistani cohorts, in which FCGR3A genotyping had previously been undertaken. Haplotype frequencies and linkage disequilibrium were estimated across the FCGR locus and a model-free analysis was performed to determine association with RA. This was followed by regression analysis, allowing for phase uncertainty, to identify the particular haplotype(s) that influences disease risk. Our results reveal that FCGR2A, FCGR2B and FCGR3B were not associated with RA. The haplotype with the strongest association with RA susceptibility was the FCGR3A-FCGR3B 158V-NA2 haplotype (odds ratio 3.18, 95% confidence interval 1.13-8.92 [P = 0.03] for homozygotes compared with all genotypes). The association was stronger in the presence of nodules (odds ratio 5.03, 95% confidence interval 1.44-17.56; P = 0.01). This haplotype was also more common in North Indian/Pakistani RA patients than in control individuals, but not significantly so. Logistic regression analyses suggested that FCGR3A remained the most significant gene at this locus. The increased association with an FCGR3A-FCGR3B haplotype suggests that other polymorphic variants within FCGR3A or FCGR3B, or in linkage disequilibrium with this haplotype, may additionally contribute to disease pathogenesis.
• Comparative Genomic and Expression Analysis of the Conserved NTPDase Gene Family in Xenopus Genomics. Mar, 2006  |   Pubmed ID: 16380227 The purines, ATP and adenosine, are important signaling molecules in the nervous system. ATP is sequentially degraded to adenosine by the ectonucleotidase proteins. The NTPDase (or CD39) family is a subfamily of these enzymes, which consists of nine members in mammals. In Xenopus embryos, we have shown that ATP, and its antagonist adenosine, regulate the rundown of swimming and we therefore proposed that ectonucleotidase proteins are key regulators of locomotor activity. Here, we report the cloning of all nine members of the NTPDase family in Xenopus laevis and Xenopus tropicalis. Our phylogenetic analysis shows that this family is highly conserved between the frog species and also during vertebrate evolution. In the adult frog, NTPDase genes are broadly expressed. During development, all NTPDase genes, except for NTPDase8, are expressed and display a distinct specific expression pattern, suggesting potentially different functions of these proteins during embryogenesis of X. laevis.
• Pbn1p: an Essential Endoplasmic Reticulum Membrane Protein Required for Protein Processing in the Endoplasmic Reticulum of Budding Yeast Proceedings of the National Academy of Sciences of the United States of America. Jan, 2006  |   Pubmed ID: 16418276 PBN1 was identified as a gene required for production of protease B (PrB) activity in Saccharomyces cerevisiae. PBN1 encodes an endoplasmic reticulum (ER)-localized, type I membrane glycoprotein and is essential for cell viability. To study the essential function(s) of Pbn1p, we constructed a strain with PBN1 under control of the GAL promoter. Depletion of Pbn1p in this strain abrogates processing of the ER precursor forms of PrB, Gas1p, and Pho8p. Depletion of Pbn1p does not affect exit of proprotease A or procarboxypeptidase Y from the ER, indicating that Pbn1p is not required for global exit from the ER. Depleting Pbn1p leads to a significant increase in the unfolded protein response pathway, accompanied by an expansion of bulk ER membrane, indicating that there is a defect in protein folding in the ER. pbn1-1, a nonlethal allele of PBN1, displays synthetic lethality with the ero1-1 allele (ERO1 is required for oxidation in the ER) and synthetic growth defects with the cne1Delta allele (CNE1 encodes calnexin). ER-associated degradation of a lumenal substrate, CPY*, is blocked in the absence of Pbn1p. These results suggest that Pbn1p is required for proper folding and/or the stability of a subset of proteins in the ER. Thus, Pbn1p is an essential chaperone-like protein in the ER of yeast.
• Dynamic Regulation of Pro- and Anti-inflammatory Cytokines by MAPK Phosphatase 1 (MKP-1) in Innate Immune Responses Proceedings of the National Academy of Sciences of the United States of America. Feb, 2006  |   Pubmed ID: 16461893 Engagement of Toll-like receptors (TLRs) on macrophages leads to activation of the mitogen-activated protein kinases (MAPKs), which contribute to innate immune responses. MAPK activity is regulated negatively by MAPK phosphatases (MKPs). MKP-1, the founding member of this family of dual-specificity phosphatases, has been implicated in regulating lipopolysaccharide (LPS) responses, but its role in TLR-mediated immune responses in vivo has not been defined. Here, we show that mice deficient in MKP-1 were highly susceptible to endotoxic shock in vivo, associated with enhanced production of proinflammatory cytokines TNF-alpha and IL-6 and an anti-inflammatory cytokine, IL-10. We further examined the regulation and function of MKP-1 in macrophages, a major cell type involved in endotoxic shock. MKP-1 was transiently induced by TLR stimulation through pathways mediated by both myeloid differentiation factor 88 (MyD88) and TIR domain-containing adaptor inducing IFN-beta (TRIF). MKP-1 deficiency led to sustained activation of p38 MAPK and c-Jun N-terminal kinase (JNK) in LPS-treated macrophages. In response to TLR signals, MKP-1-deficient macrophages produced 5- to 10-fold higher IL-10, which could be blocked by a p38 MAPK inhibitor. Thus, p38 MAPK plays a critical role in mediating IL-10 synthesis in TLR signaling. TNF-alpha was found to be more abundant in MKP-1-deficient macrophages within 2 hours of TLR stimulation, but its production was rapidly down-regulated by IL-10. Our studies demonstrate that MKP-1 attenuates the activities of p38 MAPK and JNK to regulate both pro- and anti-inflammatory cytokines in TLR signaling. These results highlight the complex mechanisms by which the MAPKs regulate innate immunity.
• A Randomized Phase II Study of Concurrent Docetaxel Plus Vaccine Versus Vaccine Alone in Metastatic Androgen-independent Prostate Cancer Clinical Cancer Research : an Official Journal of the American Association for Cancer Research. Feb, 2006  |   Pubmed ID: 16489082 Docetaxel has activity against androgen-independent prostate cancer and preclinical studies have shown that taxane-based chemotherapy can enhance antitumor response of vaccines. The primary objective of this study was to determine if concurrent docetaxel (with dexamethasone) had any effect on generating an immune response to the vaccine. Secondary end points were whether vaccine could be given safely with docetaxel and the clinical outcome of the treatment regimen.
• Naïve T-cell Dynamics in Human Immunodeficiency Virus Type 1 Infection: Effects of Highly Active Antiretroviral Therapy Provide Insights into the Mechanisms of Naive T-cell Depletion Journal of Virology. Mar, 2006  |   Pubmed ID: 16501076 Both naïve CD4+ and naïve CD8+ T cells are depleted in individuals with human immunodeficiency virus type 1 (HIV-1) infection by unknown mechanisms. Analysis of their dynamics prior to and after highly active antiretroviral therapy (HAART) could reveal possible mechanisms of depletion. Twenty patients were evaluated with immunophenotyping, intracellular Ki67 staining, T-cell receptor excision circle (TREC) quantitation in sorted CD4 and CD8 cells, and thymic computed tomography scans prior to and approximately 6 and approximately 18 months after initiation of HAART. Naïve T-cell proliferation decreased significantly during the first 6 months of therapy (P < 0.01) followed by a slower decline. Thymic indices did not change significantly over time. At baseline, naïve CD4+ T-cell numbers were lower than naive CD8+ T-cell numbers; after HAART, a greater increase in naïve CD4+ T cells than naïve CD8+ T cells was observed. A greater relative change (n-fold) in the number of TREC+ T cells/mul than in naïve T-cell counts was observed at 6 months for both CD4+ (median relative change [n-fold] of 2.2 and 1.7, respectively; P < 0.01) and CD8+ T cell pools (1.4 and 1.2; P < 0.01). A more pronounced decrease in the proliferation than the disappearance rate of naïve T cells after HAART was observed in a second group of six HIV-1-infected patients studied by in vivo pulse labeling with bromodeoxyuridine. These observations are consistent with a mathematical model where the HIV-1-induced increase in proliferation of naïve T cells is mostly explained by a faster recruitment into memory cells.
• Facilitating and Impeding Factors for Physicians' Error Disclosure: a Structured Literature Review Joint Commission Journal on Quality and Patient Safety / Joint Commission Resources. Apr, 2006  |   Pubmed ID: 16649649 It is important for physicians to disclose medical errors to institutions (for patient safety), to colleagues (for professional learning), and to patients (as part of direct patient care), but no comprehensive review of factors that may facilitate or impede disclosure has been undertaken.
• Living in Three Dimensions: 3D Nanostructured Environments for Cell Culture and Regenerative Medicine Cell Biochemistry and Biophysics. 2006  |   Pubmed ID: 16757822 Research focused on deciphering the biochemical mechanisms that regulate cell proliferation and function has largely depended on the use of tissue culture methods in which cells are grown on two-dimensional (2D) plastic or glass surfaces. However, the flat surface of the tissue culture plate represents a poor topological approximation of the more complex three-dimensional (3D) architecture of the extracellular matrix (ECM) and the basement membrane (BM), a structurally compact form of the ECM. Recent work has provided strong evidence that the highly porous nanotopography that results from the 3D associations of ECM and BM nanofibrils is essential for the reproduction of physiological patterns of cell adherence, cytoskeletal organization, migration, signal transduction, morphogenesis, and differentiation in cell culture. In vitro approximations of these nanostructured surfaces are therefore desirable for more physiologically mimetic model systems to study both normal and abnormal functions of cells, tissues, and organs. In addition, the development of 3D culture environments is imperative to achieve more accurate cell-based assays of drug sensitivity, high-throughput drug discovery assays, and in vivo and ex vivo growth of tissues for applications in regenerative medicine.
• Prospective Analysis of Uncomplicated Bone Bruises in the Pediatric Knee Emergency Radiology. Sep, 2006  |   Pubmed ID: 16816955 To determine the incidence of uncomplicated knee bone contusions in pediatric patients. MRI studies were obtained using either high-field (1.5 T) or mid-field strength magnets (0.2-0.3 T), identifying 48 pediatric patients suitable for study. Contusion location, size, and any ligamentous or meniscal injuries were recorded. Exclusionary criteria did not include plain film findings, the interval between injury to imaging, or history of patellar dislocation [Fulkerson (2002) 30:447-456]. Uncomplicated bone bruises were those occurring in the absence of other internal derangements of the knee, such as meniscal and ligament tears. Consensus imaging findings by two reviewing radiologists revealed a 25% incidence of uncomplicated bruises (12/48 patients). These bone bruises involved the lateral and medial knee compartments 56 and 44% of the time, respectively. Bruises of the lateral compartment were larger (2.4 cm) than those found in the medial compartment (1.8 cm). Given the high incidence of symptomatic but uncomplicated contusions identified in this study of a pediatric population, we suggest appropriate joint rest and follow-up without other intervention as a primary course of treatment.
• Apoptosis Inhibition by the Human DEK Oncoprotein Involves Interference with P53 Functions Molecular and Cellular Biology. Oct, 2006  |   Pubmed ID: 16894028 The DEK proto-oncogene has been associated with human carcinogenesis-either as a fusion with the CAN nucleoporin protein or when transcriptionally upregulated. Mechanisms of intracellular DEK functions, however, have remained relatively unexplored. We have recently demonstrated that DEK expression is induced by the high-risk human papillomavirus (HPV) E7 protein in a manner which is dependent upon retinoblastoma protein function and have implicated DEK in the inhibition of cellular senescence. Additionally, overexpression of DEK resulted in significant life span extension of primary human keratinocytes. In order to determine whether DEK expression is required for cellular proliferation and/or survival, we monitored cellular responses to the knockdown of DEK in cancer and primary cells. The results indicate that DEK expression protects both HPV-positive cancer and primary human cells from apoptotic cell death. Cell death in response to DEK depletion was accompanied by increased protein stability and transcriptional activity of the p53 tumor suppressor and consequent upregulation of known p53 target genes such as p21CIP and Bax. Consistent with a possible role for p53 in DEK-mediated cell death inhibition, the p53-negative human osteosarcoma cell line SAOS-2 was resistant to the knockdown of DEK. Finally, expression of a dominant negative p53 miniprotein inhibited DEK RNA interference-induced p53 transcriptional induction, as well as cell death, thus directly implicating p53 activation in the observed apoptotic phenotype. These findings suggest a novel role for DEK in cellular survival, involving the destabilization of p53 in a manner which is likely to contribute to human carcinogenesis.
• An Empirically Derived Taxonomy of Factors Affecting Physicians' Willingness to Disclose Medical Errors Journal of General Internal Medicine. Sep, 2006  |   Pubmed ID: 16918739 Physician disclosure of medical errors to institutions, patients, and colleagues is important for patient safety, patient care, and professional education. However, the variables that may facilitate or impede disclosure are diverse and lack conceptual organization.
• Cooperative Molecular and Cellular Networks Regulate Toll-like Receptor-dependent Inflammatory Responses FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology. Oct, 2006  |   Pubmed ID: 16935934 Viral and bacterial pathogens cause inflammation via Toll-like receptor (TLR) signaling. We have shown that effective responses to LPS may depend on cooperative interactions between TLR-expressing leukocytes and TLR-negative tissue cells. The aim of this work was to determine the roles of such networks in response to agonists of TLRs associated with antiviral and autoimmune responses. The TLR3 agonist poly(I:C) activated epithelial cells, primary endothelial cells, and two types of primary human smooth muscle cells (airway [ASMC] and vascular) directly, while the TLR7/8 agonist R848 required the presence of leukocytes to activate ASMC. In keeping with these data, ASMC expressed TLR3 but not TLR7 or TLR8. Activation of ASMC by poly(I:C) induced a specific cytokine repertoire characterized by induction of CXCL10 generation and the potential to recruit mast cells. We subsequently explored the ability of TLR agonists to cooperate in the induction of inflammation. Dual stimulation with LPS and poly(I:C) caused enhanced cytokine generation from epithelial and smooth muscle cells when in the presence of leukocytes. Thus, inflammatory responses to pathogens are regulated by networks in which patterns of TLR expression and colocalization of tissue cells and leukocytes are critical.
• Pediatric Epstein-Barr Virus-Associated Encephalitis: 10-Year Review Journal of Child Neurology. May, 2006  |   Pubmed ID: 16948923 Many neurologic manifestations of Epstein-Barr virus (EBV) infection have been documented, including encephalitis, aseptic meningitis, transverse myelitis, and Guillain-Barr?yndrome. These manifestations can occur alone or coincidentally with the clinical picture of infectious mononucleosis. Since 1994, The Hospital for Sick Children has maintained a prospective registry of all children admitted with acute encephalitis. This report summarizes all cases of Epstein-Barr virus-associated encephalitis compiled from 1994 to 2003. Twenty-one (6%) of 216 children, median age 13 years (range 3-17 years), in the Encephalitis Registry were identified as having evidence of Epstein-Barr virus infection. This evidence consisted of convincing Epstein-Barr virus serology and/or positive cerebrospinal fluid polymerase chain reaction (PCR). One patient had symptoms of classic infectious mononucleosis; all others had a nonspecific prodrome, including fever (n = 17; 81%) and headache (n = 14; 66%). Slightly less than half (n = 10; 48%) had seizures and often had electroencephalograms showing a slow background (n = 12; 57%). Many demonstrated cerebrospinal fluid pleocytosis (n = 17; 81%), and 71% (n = 15) had abnormal magnetic resonance imaging findings. Two patients died, 2 suffered mild deficits, and 16 were neurologically normal at follow-up. Most patients with Epstein-Barr virus encephalitis do not show typical symptoms of infectious mononucleosis. Establishing a diagnosis of Epstein-Barr virus encephalitis can be difficult, and, consequently, a combination of serologic and molecular techniques should be used when investigating a child with acute encephalitis. Most children make full recoveries, but residual neurologic sequelae and even death can and do occur. (J Child Neurol 2006;21:384-391; DOI 10.2310/7010.2006.00114).
• Cervical Cancer and Human Papillomaviruses: Inactivation of Retinoblastoma and Other Tumor Suppressor Pathways Current Molecular Medicine. Nov, 2006  |   Pubmed ID: 17100604 Infection with human papillomaviruses (HPVs) is a major public health burden worldwide and is associated with benign and malignant lesions of the skin and genital tract. HPV causes cervical cancer, which represents the second most prevalent cancer in women worldwide. Functions of the viral oncogenes E6 and E7 are essential for carcinogenesis and for support of the viral life cycle. We will begin by discussing the relationship between HPV infection and disease, followed by a review of E6 and E7 activities and their respective cellular targets. Particular emphasis will be placed on established and newly discovered mechanisms by which E7 inhibits members of the cellular retinoblastoma protein family. We will then describe how current research links the above molecular interactions to malignant transformation as well as to aspects of the viral life cycle in vitro and in vivo. As a result of decades of intense HPV research, promising therapies to prevent infection and to treat HPV associated cancers are now on the horizon. We will conclude our review by a description of potential gene therapeutic and hormonal approaches and of new developments in the design of effective vaccines.
• What Determines Blood Vessel Structure? Genetic Prespecification Vs. Hemodynamics Physiology (Bethesda, Md.). Dec, 2006  |   Pubmed ID: 17119151 Vascular network remodeling, angiogenesis, and arteriogenesis play an important role in the pathophysiology of ischemic cardiovascular diseases and cancer. Based on recent studies of vascular network development in the embryo, several novel aspects to angiogenesis have been identified as crucial to generate a functional vascular network. These aspects include specification of arterial and venous identity in vessels and network patterning. In early embryogenesis, vessel identity and positioning are genetically hardwired and involve neural guidance genes expressed in the vascular system. We demonstrated that, during later stages of embryogenesis, blood flow plays a crucial role in regulating vessel identity and network remodeling. The flow-evoked remodeling process is dynamic and involves a high degree of vessel plasticity. The open question in the field is how genetically predetermined processes in vessel identity and patterning balance with the contribution of blood flow in shaping a functional vascular architecture. Although blood flow is essential, it remains unclear to what extent flow is able to act on the developing cardiovascular system. There is significant evidence that mechanical forces created by flowing blood are biologically active within the embryo and that the level of mechanical forces and the type of flow patterns present in the embryo are able to affect gene expression. Here, we highlight the pivotal role for blood flow and physical forces in shaping the cardiovascular system.
• Aortic Arch Atheroma International Journal of Stroke : Official Journal of the International Stroke Society. May, 2006  |   Pubmed ID: 18706048 Severe atheroma of the aortic arch has now been established as an important risk factor and mechanism for stroke and peripheral embolism. The odds ratio for stroke or peripheral embolism in patients with severe arch atheroma is greater than four, and for mobile atheroma it is greater than 12. The prevalence of severe arch atheroma among patients presenting with acute ischaemic stroke, at over 20%, is in the same order as that of atrial fibrillation and carotid atherosclerosis. In patients with ischaemic stroke for which no cause has been identified, it is reasonable to determine as to whether they have severe arch atheroma by performing a transoesophageal echocardiogram. Recurrent stroke is common in patients with aortic arch atheroma that are thicker than 4 mm or with mobile components, particularly in the elderly, cigarette smokers, and those with hypertension or diabetes. Patients found to have severe atheroma are at high risk of recurrent events (14.2% per year) and may, therefore, need an aggressive secondary prevention strategy. Currently, there is uncertainty as to what this should be, but either combination antiplatelet therapy (aspirin plus clopidogrel) or anticoagulation with warfarin (target INR 2.0-3.0) are commonly used. Which of these is most effective will be evident after the completion of the aortic arch related cerebral hazard trial.
• Factors Associated with Gastrostomy Tube Feeding in Dementia: a Structured Literature Review Alzheimer's & Dementia : the Journal of the Alzheimer's Association. Jul, 2006  |   Pubmed ID: 19595892
• Static Culture of Postimplantation Embryos for Imaging CSH Protocols. 2006  |   Pubmed ID: 22485790
• GDNF Expression During Xenopus Development Gene Expression Patterns : GEP. Jan, 2007  |   Pubmed ID: 17049928 Glial cell line-derived neurotrophic factor (GDNF) has multiple roles in kidney morphogenesis, spermatogenesis, and neurogenesis during development. In this study, we report the cloning and expression pattern of Xenopus laevis GDNF. The X. laevis GDNF cDNA sequence has a complete open reading frame of 684 bases, predicting 227 amino acid residues at the protein level. Comparison of the X. laevis GDNF amino acid sequence with those of chick, human, mouse, rat and zebrafish indicates that X. laevis GDNF has 60%-52% and 75%-62% identity over the whole amino acid sequence and for the putative mature forms, respectively. All known functional motifs of GDNF were conserved in the X. laevis sequence. Temporal expression analysis by RT-PCR indicated that GDNF transcripts were first detectable at stage 12 at a low level, and gradually increased up to stage 22. From stage 24, the expression sharply increased and continued at a similar level as development progressed. Spatial expression analysis by whole-mount in situ hybridization showed that the GDNF mRNA was predominantly detected in somites, pronephros, pharyngeal arches, epibranchial placodes, digestive tract and some of the lateral line structure. These results suggest that this X. laevis gene is the orthologue for GDNF.
• Chemical Process Based Reconstruction of Exposures for an Epidemiological Study. III. Analysis of Industrial Hygiene Samples Chemico-biological Interactions. Mar, 2007  |   Pubmed ID: 17074311 As part of an historical cohort study to investigate the mortality experience of industrial workers exposed to chloroprene (beta-CD) and other substances, all available industrial hygiene exposure monitoring data were collected and summarized. From discussions with on-site industrial hygiene personnel, it was apparent that these data were not collected for epidemiological purposes and, therefore, their use in characterization of exposures was problematic as the data mostly pertained to samples collected to investigate the performance of specific tasks. These data were, however, informative for validating the exposure modeling process used to estimate historical exposures. The data summarized below clearly indicate that exposures to beta-CD were lowered across the time period of this study. Typically, the exposures recorded were less than the occupational exposure limits of the periods in which the exposures were recorded. Additionally, exposure measurements recorded in the recent past do not represent the exposure actually experienced by the worker as a strict personal protective equipment use program has been in place for the facilities studied since the mid-1980s.
• Cytogenetic and Molecular Studies of an Unusual Case of Multiple Primary Alveolar Rhabdomyosarcomas: Low-level Chromosomal Instability and Reciprocal Translocation T(6;11) Experimental and Molecular Pathology. Feb, 2007  |   Pubmed ID: 17097083 Cytogenetic and molecular studies have shown that approximately 80% of cases of alveolar rhabdomyosarcoma (ARMS) have consistent chromosomal translocation of either t(2;13) or t(1;13), resulting in either PAX3-FKHR or PAX7-FKHR gene fusions. However, 20% of the cases diagnosed histologically are negative for these fusion genes. The clinical and pathological properties of the so-called fusion gene negative tumors remain to be defined. We present an unusual case of a 7-year-old boy who developed three separate primary ARMS over a 5-year period, with the first tumor diagnosed at the age of 12 months. The tumors were negative for the characteristic translocations, t(2;13) or t(1;13), but showed evidence of low-level chromosomal instability and a reciprocal chromosomal translocation t(6;11)(q27;q13). PCR amplification of the p53 gene, exons 2-11, followed by DNA sequencing did not detect any germline p53 mutation. These clinical and cytogenetic features have not been reported previously in ARMS. The findings suggest that cytogenetic abnormalities of chromosome 6 may be associated with the development of early onset multiple ARMS in a subgroup of pediatric patients as seen in this case.
• Chordin Affects Pronephros Development in Xenopus Embryos by Anteriorizing Presomitic Mesoderm Developmental Dynamics : an Official Publication of the American Association of Anatomists. Jan, 2007  |   Pubmed ID: 17106888 Spemann's organizer emits signals that pattern the mesodermal germ layer during Xenopus embryogenesis. In a previous study, we demonstrated that FGFR1 activity within the organizer is required for the production of both the somitic muscle- and pronephros-patterning signals by the organizer and the expression of chordin, an organizer-specific secreted protein (Mitchell and Sheets [2001] Dev. Biol. 237:295-305). Studies from others in both chicken and Xenopus embryos provide compelling evidence that pronephros forms by means of secondary induction signals emitted from anterior somites (Seufert et al. [1999] Dev. Biol. 215:233-242; Mauch et al. [2000] Dev. Biol. 220:62-75). Here we provide several lines of evidence in support of the hypothesis that chordin influences pronephros development by directing the formation of anterior somites. Chordin mRNA was absent in ultraviolet (UV) -irradiated embryos lacking pronepheros (average DAI2). Furthermore, ectopic expression of chordin in embryos and in tissue explants leads to the formation of anterior somites and pronephros. In these experiments, pronephros was only observed in association with muscle. Chordin diverted somatic muscle cells to more anterior positions within the somite file in chordin-induced secondary trunks and induced the expression of the anterior myogenic gene myf5. Finally, depletion of chordin mRNA with DEED antisense oligonucleotides substantially reduced somitic muscle and pronephric tubule and duct formation in whole embryos. These data and previous studies on ectoderm and endoderm (Sasai et al. [1995] Nature 377:757) support the idea that chordin functions as an anteriorizing signal in patterning the germ layers during vertebrate embryogenesis. Our data support the hypothesis that chordin directs the formation of anterior somites that in turn are necessary for pronephros development.
• Routine Interval Computed Tomography to Detect New Soft-tissue Disease Might Be Unnecessary in Patients with Androgen-independent Prostate Cancer and Metastasis Only to Bone BJU International. Mar, 2007  |   Pubmed ID: 17155971 To identify patients with androgen-independent prostate cancer (AIPC) with bone metastasis and no soft-tissue metastases at the time of protocol enrollment, and analyse their disease progression by computed tomography (CT), bone scan, and prostate-specific antigen (PSA) level to determine the utility of routine interval CT in such patients.
• The Incidence and Natural History of Osteonecrosis in HIV-infected Adults Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America. Mar, 2007  |   Pubmed ID: 17278070 Osteonecrosis is increasingly recognized as a debilitating complication of human immunodeficiency virus (HIV) infection, but the natural history has not been well described. We previously documented a high prevalence (4.4%) of magnetic resonance imaging (MRI)-documented osteonecrosis of the hip in a cohort of 339 asymptomatic HIV-infected patients. The present study was designed to determine the incidence of newly diagnosed osteonecrosis in this cohort and to describe the natural history of osteonecrosis in HIV-infected patients.
• Association Between Physical Activity, Adiposity, and Lipid Abnormalities in Children with Familial Hyperlipidemia European Journal of Cardiovascular Prevention and Rehabilitation : Official Journal of the European Society of Cardiology, Working Groups on Epidemiology & Prevention and Cardiac Rehabilitation and Exercise Physiology. Feb, 2007  |   Pubmed ID: 17301628 The benefits of physical activity in children have been studied extensively; however, its role in children with familial hyperlipidemia (FH) is unknown.
• Pre-clinical and Clinical Evaluation of Estramustine, Docetaxel and Thalidomide Combination in Androgen-independent Prostate Cancer BJU International. May, 2007  |   Pubmed ID: 17437439 To evaluate the combination of docetaxel plus estramustine (which prolongs survival in patients with androgen-independent prostate cancer, AIPC), and thalidomide (that also adds to docetaxel activity), both pre-clinically and clinically in AIPC.
• Disclosing Medical Errors to Patients: Attitudes and Practices of Physicians and Trainees Journal of General Internal Medicine. Jul, 2007  |   Pubmed ID: 17473944 Disclosing errors to patients is an important part of patient care, but the prevalence of disclosure, and factors affecting it, are poorly understood.
• Ultrasonographic Examination of the Synovial Fold of the Radiohumeral Joint Journal of Shoulder and Elbow Surgery / American Shoulder and Elbow Surgeons ... [et Al.]. Sep-Oct, 2007  |   Pubmed ID: 17507252 This report describes the anatomy of the synovial fold of the radiohumeral joint and assesses its visibility by ultrasonography. Forty-nine fresh cadaver radiohumeral joints were examined by ultrasonography before and after intraarticular saline injection and then dissected. Digital photos were taken before and after the joint capsule was excised. The relative coverage of the radial head by the fold was calculated. Synovial folds were observed in all specimens. Forty-three had anterior and posterior lobes. The synovial fold covered an average of 28% of the radiocapitellar joint surface of the radial head. The sensitivity of the ultrasonography was 81%, 46%, and 85% from the anterior, lateral, and posterior aspects of the radiohumeral joint, respectively. Intraarticular saline injection improved the sensitivity to 96%, 67%, and 94%, respectively. The synovial fold is a consistent anatomic structure, and ultrasonography can be a useful preoperative diagnostic tool.
• Genetic Identity Determines Risk of Post-settlement Mortality of a Marine Fish Ecology. May, 2007  |   Pubmed ID: 17536412 Longitudinal sampling of four cohorts of Neopomacentrus filamentosus, a common tropical damselfish from Dampier Archipelago, Western Australia, revealed the evolution of size structure after settlement. Light traps collected premetamorphic individuals from the water column ("settlers") to establish a baseline for each cohort. Subsequently, divers collected benthic juveniles ("recruits") at 1-3-month intervals to determine the relative impacts of post-settlement mortality during the first three months. Growth trajectories for individual fish were back-calculated from otolith records and compared with nonlinear mixed-effects models. Size-selective mortality was detected in all cohorts with the loss of smaller, slower growing individuals. Three months after settlement, recruits showed significantly faster growth as juveniles, faster growth as larvae, and larger sizes as hatchlings. The timing and intensity of post-settlement selection differed among cohorts and was correlated with density at settlement. The cohort with the greatest initial abundance experienced the strongest selective mortality, with most of this mortality occurring between one and two months after settlement when juveniles began foraging at higher positions in the water column. Significant genetic structure was found between settlers and three-month-old recruits in this cohort as a result of natural selection that changed the frequency of mtDNA haplotypes measured at the control region. The extent of this genetic difference was enlarged or reduced by artificially manipulating the intensity of size-based selection, thus establishing a link between phenotype and haplotype. Sequence variation in the control region of the mitochondrial genome has been linked to mitochondrial efficiency and weight gain in other studies, which provides a plausible explanation for the patterns observed here.
• Optimising the Provision of Human Milk for Preterm Infants Archives of Disease in Childhood. Fetal and Neonatal Edition. Jul, 2007  |   Pubmed ID: 17585091
• Regulation of SRC-3 Intercompartmental Dynamics by Estrogen Receptor and Phosphorylation Molecular and Cellular Biology. Oct, 2007  |   Pubmed ID: 17646391 The steroid receptor coactivator 3 gene (SRC-3) (AIB1/ACTR/pCIP/RAC3/TRAM1) is a p160 family transcription coactivator and a known oncogene. Despite its importance, the functional regulation of SRC-3 remains poorly understood within a cellular context. Using a novel combination of live-cell, high-throughput, and fluorescent microscopy, we report SRC-3 to be a nucleocytoplasmic shuttling protein whose intracellular mobility, solubility, and cellular localization are regulated by phosphorylation and estrogen receptor alpha (ERalpha) interactions. We show that both chemical inhibition and small interfering RNA reduction of the mitogen-activated protein kinase/extracellular signal-regulated kinase 1/2 (MEK1/2) pathway induce a cytoplasmic shift in SRC-3 localization, whereas stimulation by epidermal growth factor signaling enhances its nuclear localization by inducing phosphorylation at T24, S857, and S860, known participants in the phosphocode that regulates SRC-3 activity. Accordingly, the cytoplasmic localization of a nonphosphorylatable SRC-3 mutant further supported these results. In the presence of ERalpha, U0126 also dramatically reduces (i) ligand-dependent colocalization of SRC-3 and ERalpha, (ii) the formation of ER-SRC-3 complexes in cell lysates, and (iii) SRC-3 targeting to a visible, ERalpha-occupied and -regulated prolactin promoter array. Taken together, these results indicate that phosphorylation coordinates SRC-3 coactivator function by linking the probabilistic formation of transient nuclear receptor-coactivator complexes with its molecular dynamics and cellular compartmentalization. Technically and conceptually, these findings have a new and broad impact upon evaluating mechanisms of action of gene regulators at a cellular system level.
• Clinical Safety of a Viral Vector Based Prostate Cancer Vaccine Strategy The Journal of Urology. Oct, 2007  |   Pubmed ID: 17707059 The primary objective of this phase I study was to evaluate the clinical safety of a vaccine using recombinant vaccinia virus (prime) and recombinant fowlpox virus (boost) in combination with granulocyte-macrophage colony-stimulating factor in patients with prostate cancer. The vaccines contained transgenes for prostate specific antigen, a triad of co-stimulatory molecules and a tumor antigen whose amino acid sequence had been modified to enhance its immunogenicity. Secondary end points were immunological and clinical responses, changes in prostate specific antigen velocity, and the kinetics of vaccinia virus clearance from the vaccination site, serum, peripheral blood mononuclear cells, urine and saliva.
• Extreme Obesity Among Children in Mexico The Journal of Pediatrics. Sep, 2007  |   Pubmed ID: 17719922
• Vascular Remodeling of the Mouse Yolk Sac Requires Hemodynamic Force Development (Cambridge, England). Sep, 2007  |   Pubmed ID: 17720695 The embryonic heart and vessels are dynamic and form and remodel while functional. Much has been learned about the genetic mechanisms underlying the development of the cardiovascular system, but we are just beginning to understand how changes in heart and vessel structure are influenced by hemodynamic forces such as shear stress. Recent work has shown that vessel remodeling in the mouse yolk sac is secondarily effected when cardiac function is reduced or absent. These findings indicate that proper circulation is required for vessel remodeling, but have not defined whether the role of circulation is to provide mechanical cues, to deliver oxygen or to circulate signaling molecules. Here, we used time-lapse confocal microscopy to determine the role of fluid-derived forces in vessel remodeling in the developing murine yolk sac. Novel methods were used to characterize flows in normal embryos and in embryos with impaired contractility (Mlc2a(-/-)). We found abnormal plasma and erythroblast circulation in these embryos, which led us to hypothesize that the entry of erythroblasts into circulation is a key event in triggering vessel remodeling. We tested this by sequestering erythroblasts in the blood islands, thereby lowering the hematocrit and reducing shear stress, and found that vessel remodeling and the expression of eNOS (Nos3) depends on erythroblast flow. Further, we rescued remodeling defects and eNOS expression in low-hematocrit embryos by restoring the viscosity of the blood. These data show that hemodynamic force is necessary and sufficient to induce vessel remodeling in the mammalian yolk sac.
• Inhibition of P-glycoprotein and Multidrug Resistance-associated Proteins Modulates the Intracellular Concentration of Lopinavir in Cultured CD4 T Cells and Primary Human Lymphocytes The Journal of Antimicrobial Chemotherapy. Nov, 2007  |   Pubmed ID: 17890284 HIV protease inhibitors (HPIs) are an important component of highly active antiretroviral therapy. However, the activity of drug efflux transporters, such as P-glycoprotein (P-gp) and multidrug resistance-associated proteins (MRP1/MRP2), may limit intracellular drug accumulation. Drugs that inhibit the activity of drug efflux proteins may, in combination with HPIs, enhance the clinical efficacy of the drugs.
• Purine-mediated Signalling Triggers Eye Development Nature. Oct, 2007  |   Pubmed ID: 17960245 A conserved network of eye field transcription factors (EFTFs) underlies the development of the eye in vertebrates and invertebrates. To direct eye development, Pax6, a key gene in this network, interacts with genes encoding other EFTFs such as Rx1 and Six3 (refs 4-6). However, the mechanisms that control expression of the EFTFs remain unclear. Here we show that purine-mediated signalling triggers both EFTF expression and eye development in Xenopus laevis. Overexpression of ectonucleoside triphosphate diphosphohydrolase 2 (E-NTPDase2), an ectoenzyme that converts ATP to ADP, caused ectopic eye-like structures, with occasional complete duplication of the eye, and increased expression of Pax6, Rx1 and Six3. In contrast, downregulation of endogenous E-NTPDase2 decreased Rx1 and Pax6 expression. E-NTPDase2 therefore acts upstream of these EFTFs. To test whether ADP (the product of E-NTPDase2) might act to trigger eye development through P2Y1 receptors, selective in Xenopus for ADP, we simultaneously knocked down expression of the genes encoding E-NTPDase2 and the P2Y1 receptor. This could prevent the expression of Rx1 and Pax6 and eye formation completely. We next measured ATP release in the presumptive eye field, demonstrating a transient release of ATP at a time that could plausibly trigger (once converted to ADP) expression of the EFTFs. This surprising role for transient purine-mediated signalling in eye development may be widely conserved, because alterations to the locus of E-NTPDase2 on human chromosome 9 cause severe head and eye defects, including microphthalmia. Our results suggest a new mechanism for the initiation of eye development.
• Anxa4 Genes Are Expressed in Distinct Organ Systems in Xenopus Laevis and Tropicalis But Are Functionally Conserved Organogenesis. Oct, 2007  |   Pubmed ID: 19279706 Anxa4 belongs to the multigenic annexin family of proteins which are characterized by their ability to interact with membranes in a calcium-dependent manner. Defined as a marker for polarized epithelial cells, Anxa4 is believed to be involved in many cellular processes but its functions in vivo are still poorly understood. Previously, we cloned Xanx4 in Xenopus laevis (now referred to as anxa4a) and demonstrated its role during organogenesis of the pronephros, providing the first evidence of a specific function for this protein during the development of a vertebrate. Here, we describe the strict conservation of protein sequence and functional domains of anxa4 during vertebrate evolution. We also identify the paralog of anxa4a, anxa4b and show its specific temporal and spatial expression pattern is different from anxa4a. We show that anxa4 orthologs in X. laevis and tropicalis display expression domains in different organ systems. Whilst the anxa4a gene is mainly expressed in the kidney, Xt anxa4 is expressed in the liver. Finally, we demonstrate Xt anxa4 and anxa4a can display conserved function during kidney organogenesis, despite the fact that Xt anxa4 transcripts are not expressed in this domain. This study highlights the divergence of expression of homologous genes during Xenopus evolution and raises the potential problems of using X. tropicalis promoters in X. laevis.
• The Spectrum of Trp- Mutants Isolated As 5-fluoroanthranilate-resistant Clones in Saccharomyces Bayanus, S. Mikatae and S. Paradoxus Yeast (Chichester, England). Jan, 2008  |   Pubmed ID: 17924454 5-Fluoroanthranilic acid (FAA)-resistant mutants were selected in homothallic diploids of three Saccharomyces species, taking care to isolate mutants of independent origin. Mutations were assigned to complementation groups by interspecific complementation with S. cerevisiae tester strains. In all three species, trp3, trp4 and trp5 mutants were recovered. trp1 mutants were also recovered if the selection was imposed on a haploid strain. Thus, FAA selection may be more generally applicable than was previously described.
• Intraobserver and Interobserver Variability of Transesophageal Echocardiography in Aortic Arch Atheroma Measurement Journal of the American Society of Echocardiography : Official Publication of the American Society of Echocardiography. Feb, 2008  |   Pubmed ID: 17935947 The risk of stroke associated with aortic arch atheroma is proportional to plaque thickness, the accurate measurement of which may influence future therapeutic decisions. Transesophageal echocardiography is the procedure of choice for plaque measurement. This study examines the reproducibility of such measurements within the context of the Aortic Arch Related Cerebral Hazard Trial of antithrombotic therapy.
• A Phase II Clinical Trial of Sorafenib in Androgen-independent Prostate Cancer Clinical Cancer Research : an Official Journal of the American Association for Cancer Research. Jan, 2008  |   Pubmed ID: 18172272 To determine if sorafenib is associated with a 4-month probability of progression-free survival, which is consistent with 50%, as determined by clinical, radiographic, and prostate-specific antigen (PSA) criteria in patients with metastatic androgen-independent prostate cancer (AIPC).
• Reporting Medical Errors to Improve Patient Safety: a Survey of Physicians in Teaching Hospitals Archives of Internal Medicine. Jan, 2008  |   Pubmed ID: 18195194 Collecting data on medical errors is essential for improving patient safety, but factors affecting error reporting by physicians are poorly understood.
• A Functional Screen for Genes Involved in Xenopus Pronephros Development Mechanisms of Development. Jul, 2008  |   Pubmed ID: 18472403 In Xenopus, the pronephros is the functional larval kidney and consists of two identifiable components; the glomus, the pronephric tubules, which can be divided into four separate segments, based on marker gene expression. The simplicity of this organ, coupled with the fact that it displays the same basic organization and function as more complex mesonephros and metanephros, makes this an attractive model to study vertebrate kidney formation. In this study, we have performed a functional screen specifically to identify genes involved in pronephros development in Xenopus. Gain-of-function screens are performed by injecting mRNA pools made from a non-redundant X. tropicalis full-length plasmid cDNA library into X. laevis eggs, followed by sib-selection to identify the single clone that caused abnormal phenotypes in the pronephros. Out of 768 egg and gastrula stage cDNA clones, 31 genes, approximately 4% of the screened clones, affected pronephric marker expression examined by whole mount in situ hybridization or antibody staining. Most of the positive clones had clear expression patterns in pronephros and predicted/established functions highly likely to be involved in developmental processes. In order to carry out a more detailed study, we selected Sox7, Cpeb3, P53csv, Mecr and Dnajc15, which had highly specific expression patterns in the pronephric region. The over-expression of these five selected clones indicated that they caused pronephric abnormalities with different temporal and spatial effects. These results suggest that our strategy to identify novel genes involved in pronephros development was highly successful, and that this strategy is effective for the identification of novel genes involved in late developmental events.
• Separating Genetic and Hemodynamic Defects in Neuropilin 1 Knockout Embryos Development (Cambridge, England). Aug, 2008  |   Pubmed ID: 18550715 Targeted inactivation of genes involved in murine cardiovascular development frequently leads to abnormalities in blood flow. As blood fluid dynamics play a crucial role in shaping vessel morphology, the presence of flow defects generally prohibits the precise assignment of the role of the mutated gene product in the vasculature. In this study, we show how to distinguish between genetic defects caused by targeted inactivation of the neuropilin 1 (Nrp1) receptor and hemodynamic defects occurring in homozygous knockout embryos. Our analysis of a Nrp1 null allele bred onto a C57BL/6 background shows that vessel remodeling defects occur concomitantly with the onset of blood flow and cause death of homozygous mutants at E10.5. Using mouse embryo culture, we establish that hemodynamic defects are already present at E8.5 and continuous circulation is never established in homozygous mutants. The geometry of yolk sac blood vessels is altered and remodeling into yolk sac arteries and veins does not occur. To separate flow-induced deficiencies from those caused by the Nrp1 mutation, we arrested blood flow in cultured wild-type and mutant embryos and followed their vascular development. We find that loss of Nrp1 function rather than flow induces the altered geometry of the capillary plexus. Endothelial cell migration, but not replication, is altered in Nrp1 mutants. Gene expression analysis of endothelial cells isolated from freshly dissected wild-type and mutants and after culture in no-flow conditions showed down-regulation of the arterial marker genes connexin 40 and ephrin B2 related to the loss of Nrp1 function. This method allows genetic defects caused by loss-of-function of a gene important for cardiovascular development to be isolated even in the presence of hemodynamic defects.
• The Lmx1b Gene is Pivotal in Glomus Development in Xenopus Laevis Developmental Biology. Oct, 2008  |   Pubmed ID: 18687324 We have previously shown that lmx1b, a LIM homeodomain protein, is expressed in the pronephric glomus. We now show temporal and spatial expression patterns of lmx1b and its potential binding partners in both dissected pronephric anlagen and in individual dissected components of stage 42 pronephroi. Morpholino oligonucleotide knock-down of lmx1b establishes a role for lmx1b in the development of the pronephric components. Depletion of lmx1b results in the formation of a glomus with reduced size. Pronephric tubules were also shown to be reduced in structure and/or coiling whereas more distal tubule structure was unaffected. Over-expression of lmx1b mRNA resulted in no significant phenotype. Given that lmx1b protein is known to function as a heterodimer, we have over-expressed lmx1b mRNA alone or in combination with potential interacting molecules and analysed the effects on kidney structures. Phenotypes observed by over-expression of lim1 and ldb1 are partially rescued by co-injection with lmx1b mRNA. Animal cap experiments confirm that co-injection of lmx1b with potential binding partners can up-regulate pronephric molecular markers suggesting that lmx1b lies upstream of wt1 in the gene network controlling glomus differentiation. This places lmx1b in a genetic hierarchy involved in pronephros development and suggests that it is the balance in levels of binding partners together with restricted expression domains of lmx1b and lim1 which influences differentiation into glomus or tubule derivatives in vivo.
• A Rainbow of Fluoromodules: a Promiscuous ScFv Protein Binds to and Activates a Diverse Set of Fluorogenic Cyanine Dyes Journal of the American Chemical Society. Sep, 2008  |   Pubmed ID: 18761447 Combined magnetic and fluorescence cell sorting were used to select Fluorogen Activating Proteins (FAPs) from a yeast surface-displayed library for binding to the fluorogenic cyanine dye Dimethyl Indole Red (DIR). Several FAPs were selected that bind to the dye with low nanomolar Kd values and enhance fluorescence more than 100-fold. One of these FAPs also exhibits considerable promiscuity, binding with high affinity to several other fluorogenic cyanine dyes with emission wavelengths covering most of the visible and near-IR regions of the spectrum. This significantly expands the number and wavelength range of scFv-based fluoromodules.
• Comparative Phylogeography of Two Seastars and Their Ectosymbionts Within the Coral Triangle Molecular Ecology. Dec, 2008  |   Pubmed ID: 19067797 Repeated exposure and flooding of the Sunda and Sahul shelves during Pleistocene sea-level fluctuations is thought to have contributed to the isolation and diversification of sea-basin populations within the Coral Triangle. This hypothesis has been tested in numerous phylogeographical studies, recovering an assortment of genetic patterns that the authors have generally attributed to differences in larval dispersal capability or adult habitat specificity. This study compares phylogeographical patterns from mitochondrial COI sequences among two co-distributed seastars that differ in their adult habitat and dispersal ability, and two seastar ectosymbionts that differ in their degree of host specificity. Of these, only the seastar Linckia laevigata displayed a classical pattern of Indian-Pacific divergence, but with only moderate genetic structure (PhiCT = 0.067). In contrast, the seastarProtoreaster nodosus exhibited strong structure (PhiCT = 0.23) between Teluk Cenderawasih and the remainder of Indonesia, a pattern of regional structure that was echoed in L. laevigata (PhiCT = 0.03) as well as its obligate gastropod parasite Thyca crystallina (PhiCT = 0.04). The generalist commensal shrimp, Periclimenes soror showed little genetic structuring across the Coral Triangle. Despite species-specific phylogeographical patterns, all four species showed departures from neutrality that are consistent with massive range expansions onto the continental shelves as the sea levels rose, and that date within the Pleistocene epoch.Our results suggest that habitat differences may affect the manner in which species responded to Pleistocene sea-level fluctuations, shaping contemporary patterns of genetic structure and diversity.
• Requirement of Wnt/beta-catenin Signaling in Pronephric Kidney Development Mechanisms of Development. Mar-Apr, 2009  |   Pubmed ID: 19100832 The pronephric kidney controls water and electrolyte balance during early fish and amphibian embryogenesis. Many Wnt signaling components have been implicated in kidney development. Specifically, in Xenopus pronephric development as well as the murine metanephroi, the secreted glycoprotein Wnt-4 has been shown to be essential for renal tubule formation. Despite the importance of Wnt signals in kidney organogenesis, little is known of the definitive downstream signaling pathway(s) that mediate their effects. Here we report that inhibition of Wnt/beta-catenin signaling within the pronephric field of Xenopus results in significant losses to kidney epithelial tubulogenesis with little or no effect on adjoining axis or somite development. We find that the requirement for Wnt/beta-catenin signaling extends throughout the pronephric primordium and is essential for the development of proximal and distal tubules of the pronephros as well as for the development of the duct and glomus. Although less pronounced than effects upon later pronephric tubule differentiation, inhibition of the Wnt/beta-catenin pathway decreased expression of early pronephric mesenchymal markers indicating it is also needed in early pronephric patterning. We find that upstream inhibition of Wnt/beta-catenin signals in zebrafish likewise reduces pronephric epithelial tubulogenesis. We also find that exogenous activation of Wnt/beta-catenin signaling within the Xenopus pronephric field results in significant tubulogenic losses. Together, we propose Wnt/beta-catenin signaling is required for pronephric tubule, duct and glomus formation in Xenopus laevis, and this requirement is conserved in zebrafish pronephric tubule formation.
• Final Analysis of a Phase II Trial Using Sorafenib for Metastatic Castration-resistant Prostate Cancer BJU International. Jun, 2009  |   Pubmed ID: 19154507 To determine if sorafenib is associated with an improved 4-month probability of progression-free survival, using radiographic and clinical criteria alone, in patients with metastatic castration-resistant prostate cancer. Secondary endpoints included pharmacokinetics, toxicity analysis and overall survival.
• Clinical Significance of Reporting Benign-appearing Endometrial Cells in Pap Tests in Women Aged 40 Years and Over Acta Cytologica. Jan-Feb, 2009  |   Pubmed ID: 19248550 To determine the clinical significance of reporting benign-appearing endometrial cells on Pap tests from women > or = 40 years.
• What Characterizes Persons Who Do Not Report Musculoskeletal Pain? Results from a 4-year Population-based Longitudinal Study (the Epifund Study) The Journal of Rheumatology. May, 2009  |   Pubmed ID: 19369469 To identify and characterize persons in the population who do not report musculoskeletal pain.
• A Critical Appraisal of Factors Affecting the Accuracy of Results Obtained when Using Flow Cytometry in Stem Cell Investigations: Where Do You Put Your Gates? Cytometry. Part A : the Journal of the International Society for Analytical Cytology. Sep, 2009  |   Pubmed ID: 19562683 Flow cytometry is used extensively in stem cell investigations but there is wide variation in the methods used for data analysis between laboratories. Data analysis can be challenging in stem cell biology as there is often no clear distinction between positive and negative populations. We have undertaken a critical appraisal of factors that affect the accuracy of results in stem cell applications. We used mouse embryonic stem cells and determined the expression of three common antigens in stem cell investigations, namely CD15, CD184, and c-kit. We have compared different cell preparation methods and gating strategies and also evaluated the use of isotype controls and unstained cells as controls for the identification of positive populations. The use of a "doublet discriminator" using a side scatter area signal versus side scatter height signal dot plot to identify single cells for analysis increases the accuracy of results regardless of the method used to dissociate cells. Isotype controls can be helpful in mimicking cellular nonspecific binding of the experimental antibody reaction. Isotype controls behave differently on stem cells at different stages of differentiation. Analysis of a viable single cell population with careful selection of control cell populations increases the accuracy of results.
• Genetic Properties of the Maize Nested Association Mapping Population Science (New York, N.Y.). Aug, 2009  |   Pubmed ID: 19661427 Maize genetic diversity has been used to understand the molecular basis of phenotypic variation and to improve agricultural efficiency and sustainability. We crossed 25 diverse inbred maize lines to the B73 reference line, capturing a total of 136,000 recombination events. Variation for recombination frequencies was observed among families, influenced by local (cis) genetic variation. We identified evidence for numerous minor single-locus effects but little two-locus linkage disequilibrium or segregation distortion, which indicated a limited role for genes with large effects and epistatic interactions on fitness. We observed excess residual heterozygosity in pericentromeric regions, which suggested that selection in inbred lines has been less efficient in these regions because of reduced recombination frequency. This implies that pericentromeric regions may contribute disproportionally to heterosis.
• Notch Activates Wnt-4 Signalling to Control Medio-lateral Patterning of the Pronephros Development (Cambridge, England). Nov, 2009  |   Pubmed ID: 19793883 Previous studies have highlighted a role for the Notch signalling pathway during pronephrogenesis in the amphibian Xenopus laevis, and in nephron development in the mammalian metanephros, yet a mechanism for this function remains elusive. Here, we further the understanding of how Notch signalling patterns the early X. laevis pronephros anlagen, a function that might be conserved in mammalian nephron segmentation. Our results indicate that early phase pronephric Notch signalling patterns the medio-lateral axis of the dorso-anterior pronephros anlagen, permitting the glomus and tubules to develop in isolation. We show that this novel function acts through the Notch effector gene hrt1 by upregulating expression of wnt4. Wnt-4 then patterns the proximal pronephric anlagen to establish the specific compartments that span the medio-lateral axis. We also identified pronephric expression of lunatic fringe and radical fringe that is temporally and spatially appropriate for a role in regulating Notch signalling in the dorso-anterior region of the pronephros anlagen. On the basis of these results, along with data from previous publications, we propose a mechanism by which the Notch signalling pathway regulates a Wnt-4 function that patterns the proximal pronephric anlagen.
• Breast-specific Gamma-imaging: Molecular Imaging of the Breast Using 99mTc-sestamibi and a Small-field-of-view Gamma-camera Journal of Nuclear Medicine Technology. Dec, 2009  |   Pubmed ID: 19914975 Breast-specific gamma-imaging (BSGI), also known as molecular breast imaging, is breast scintigraphy using a small-field-of-view gamma-camera and (99m)Tc-sestamibi. There are many different types of breast cancer, and many have characteristics making them challenging to detect by mammography and ultrasound. BSGI is a cost-effective, highly sensitive and specific technique that complements other imaging modalities currently being used to identify malignant lesions in the breast. Using the current Society of Nuclear Medicine guidelines for breast scintigraphy, Legacy Good Samaritan Hospital began conducting BSGI, breast scintigraphy with a breast-optimized gamma-camera. In our experience, optimal imaging has been conducted in the Breast Center by a nuclear medicine technologist. In addition, the breast radiologists read the BSGI images in correlation with the mammograms, ultrasounds, and other imaging studies performed. By modifying the current Society of Nuclear Medicine protocol to adapt it to the practice of breast scintigraphy with these new systems and by providing image interpretation in conjunction with the other breast imaging studies, our center has found BSGI to be a valuable adjunctive procedure in the diagnosis of breast cancer. The development of a small-field-of-view gamma-camera, designed to optimize breast imaging, has resulted in improved detection capabilities, particularly for lesions less than 1 cm. Our experience with this procedure has proven to aid in the clinical work-up of many of our breast patients. After reading this article, the reader should understand the history of breast scintigraphy, the pharmaceutical used, patient preparation and positioning, imaging protocol guidelines, clinical indications, and the role of breast scintigraphy in breast cancer diagnosis.
• Managing MRSA in Retail Health Advance for Nurse Practitioners. Jul, 2009  |   Pubmed ID: 19999409
• Normal Levels of P27 Are Necessary for Somite Segmentation and Determining Pronephric Organ Size Organogenesis. Oct, 2009  |   Pubmed ID: 20539739 The Xenopus laevis cyclin dependent kinase inhibitor p27(Xic1) has been shown to be involved in exit from the cell cycle and differentiation of cells into a quiescent state in the nervous system, muscle tissue, heart and retina. We show that p27(Xic1) is expressed in the developing kidney in the nephrostomal regions. Using overexpression and morpholino oligonucleotide (MO) knock-down approaches we show normal levels of p27(Xic1) regulate pronephros organ size by regulating cell cycle exit. Knock-down of p27(Xic1) expression using a MO prevented myogenesis, as previously reported; an effect that subsequently inhibits pronephrogenesis. Furthermore, we show that normal levels of p27(Xic1) are required for somite segmentation also through its cell cycle control function. Finally, we provide evidence to suggest correct paraxial mesoderm segmentation is not necessary for pronephric induction in the intermediate mesoderm. These results indicate novel developmental roles for p27(Xic1), and reveal its differentiation function is not universally utilised in all developing tissues.
• Xenopus Wnt11b is Identified As a Potential Pronephric Inducer Developmental Dynamics : an Official Publication of the American Association of Anatomists. Jan, 2010  |   Pubmed ID: 19582868 In this study, we aimed to establish if known wnt signaling molecules could be responsible for inducing early pronephros specification, using a novel and effective in vitro bioassay in Xenopus embryos. Anterior somites have the unique biological activity to signal to unspecified intermediate mesoderm to induce pronephros formation in Xenopus embryos. We have used a molecular candidate gene approach to analyze both canonical and noncanonical wnt expression in isolated anterior and posterior somites and dissected presumptive pronephros, pronephric anlagen, and pronephros from stage 12.5-35 embryos. We have identified potential candidate wnt genes expressed in the right time and place to specify pronephric development. These candidates were then directly tested in an in vitro pronephrogenesis assay based on Holtfreter sandwich cultures. Results revealed that noncanonical wnt11b and wnt11 can induce pronephros formation in vitro. Loss-of-function experiments confirmed that these genes are necessary for normal pronephros development.
• Ectophosphodiesterase/nucleotide Phosphohydrolase (Enpp) Nucleotidases: Cloning, Conservation and Developmental Restriction The International Journal of Developmental Biology. 2010  |   Pubmed ID: 19598106 Ectonucleotidase proteins occupy a central role in purine signalling regulation by sequentially hydrolysing ATP to ADP and to adenosine. The ENPP ( or PDNP) gene family, which encodes ectophosphodiesterase/nucleotide phosphohydrolases, is a subfamily of these enzymes, which consists of 7 members in mammals. These proteins catalyse the generation of bioactive lipids, placing the ENPP enzymes as key regulators of major physiological signalling pathways and also important players in several pathological conditions. Here we report the cloning of all the members, except enpp5, of the enpp family in Xenopus laevis and tropicalis. Phylogenetic analyses demonstrate the high level of conservation of these proteins between amphibian and other vertebrate species. During development and in the adult frog, each gene displays a distinct specific expression pattern, suggesting potentially different functions for these proteins during amphibian embryogenesis. This is the first complete developmental analysis of gene expression of this gene family in vertebrates.
• Immunologic and Prognostic Factors Associated with Overall Survival Employing a Poxviral-based PSA Vaccine in Metastatic Castrate-resistant Prostate Cancer Cancer Immunology, Immunotherapy : CII. May, 2010  |   Pubmed ID: 19890632 A concurrent multicenter, randomized Phase II trial employing a recombinant poxviral vaccine provided evidence of enhanced median overall survival (OS) (p = 0.0061) in patients with metastatic castrate-resistant prostate cancer (mCRPC). The study reported here employed the identical vaccine in mCRPC to investigate the influence of GM-CSF with vaccine, and the influence of immunologic and prognostic factors on median OS. Thirty-two patients were vaccinated once with recombinant vaccinia containing the transgenes for prostate-specific antigen (PSA) and three costimulatory molecules. Patients received boosters with recombinant fowlpox containing the same four transgenes. Twelve of 32 patients showed declines in serum PSA post-vaccination and 2/12 showed decreases in index lesions. Median OS was 26.6 months (predicted median OS by the Halabi nomogram was 17.4 months). Patients with greater PSA-specific T-cell responses showed a trend (p = 0.055) toward enhanced survival. There was no difference in T-cell responses or survival in cohorts of patients receiving GM-CSF versus no GM-CSF. Patients with a Halabi predicted survival of or =18 months (median predicted survival 20.9 months) will meet or exceed 37.3 months, with 12/15 patients living longer than predicted (p = 0.035). Treg suppressive function was shown to decrease following vaccine in patients surviving longer than predicted, and increase in patients surviving less than predicted. This hypothesis-generating study provides evidence that patients with more indolent mCRPC (Halabi predicted survival > or =18 months) may best benefit from vaccine therapy.
• Building Local Community Commitment to Wetlands Restoration: a Case Study of the Cache River Wetlands in Southern Illinois, USA Environmental Management. Apr, 2010  |   Pubmed ID: 20127327 Natural resource professionals are increasingly faced with the challenges of cultivating community-based support for wetland ecosystem restoration. While extensive research efforts have been directed toward understanding the biophysical dimensions of wetland conservation, the literature provides less guidance on how to successfully integrate community stakeholders into restoration planning. Therefore, this study explores the social construction of wetlands locally, and community members' perceptions of the wetland restoration project in the Cache River Watershed of southern Illinois, where public and private agencies have partnered together to implement a large-scale wetlands restoration project. Findings illustrate that the wetlands hold diverse and significant meanings to community members and that community members' criteria for project success may vary from those identified by project managers. The case study provides managers with strategies for building community commitment such as engaging local citizens in project planning, minimizing local burdens, maximizing local benefits, and reducing uncertainty.
• Investigation of a Problem with External Quality Assurance Samples for Faecal Occult Blood Annals of Clinical Biochemistry. Mar, 2010  |   Pubmed ID: 20150216 It is vital that laboratories participate in External Quality Assurance (EQA) programmes, but results from such schemes do not necessarily help ensure assays are 'in control'. We describe a series of experiments undertaken to explore apparent poor performance in an occult blood EQA scheme that manifested as a series of false-negatives. As a consequence of our laboratory misclassifying some EQA samples, we decided to design a simple sensitivity study that would reaffirm confidence in our testing procedures.
• A Protocol for Imaging Axillary Lymph Nodes in Patients Undergoing Breast-specific Gamma-imaging Journal of Nuclear Medicine Technology. Mar, 2010  |   Pubmed ID: 20159928 The development of small-field-of-view, breast-optimized, gamma-camera designed for breast scintigraphy has resulted in improved breast lesion detection-particularly for lesions smaller than 1 cm. However, unlike with the standard gamma-camera, these images do not include the axilla within the field of view.
• The Culture of Mouse Embryonic Stem Cells and Formation of Embryoid Bodies Methods in Molecular Biology (Clifton, N.J.). 2010  |   Pubmed ID: 20204616 Embryonic stem (ES) cells are pluripotent cells isolated from the inner cell mass of the pre-implantation blastocyst. They have the capacity to undergo indefinite rounds of self-renewing cell division and differentiate into all the cell lineages of the developing embryo. In suspension culture, ES cells will differentiate into aggregates known as embryoid bodies in a manner similar to the early embryo. This culture system therefore provides a useful model to study the relatively inaccessible stages of mammalian development. We describe methods for the routine maintenance of mouse embryonic stem cells in culture, assays of stem cell self-renewal potential in monolayer culture and the generation of embryoid bodies to study differentiation pathways.
• Concentration-dependent Effects and Intracellular Accumulation of HIV Protease Inhibitors in Cultured CD4 T Cells and Primary Human Lymphocytes The Journal of Antimicrobial Chemotherapy. May, 2010  |   Pubmed ID: 20237075 The intracellular and plasma concentrations of HIV protease inhibitors (HPIs) vary widely in vivo. It is unclear whether there is a concentration-dependent effect of HPIs such that at increasing concentration they may either block their own efflux (leading to 'autoboosting') or influx (leading to saturability/decreased intracellular accumulation).
• Phase II Trial of Bevacizumab, Thalidomide, Docetaxel, and Prednisone in Patients with Metastatic Castration-resistant Prostate Cancer Journal of Clinical Oncology : Official Journal of the American Society of Clinical Oncology. Apr, 2010  |   Pubmed ID: 20308663 PURPOSE We previously demonstrated that thalidomide appears to add to the activity of docetaxel in metastatic castration-resistant prostate cancer (CRPC). Phase II studies combining docetaxel with bevacizumab have had substantial antitumor activity. We hypothesized that the combination of docetaxel plus these antiangiogenic drugs with different targets would have substantial clinical activity. To explore safety and efficacy, this was tested in mice and in human patients. PATIENTS AND METHODS Preclinical efficacy of the combination therapy was evaluated in PC3 xenograft mice. Sixty patients with progressive metastatic CRPC received intravenous docetaxel and bevacizumab plus oral thalidomide and prednisone. The primary end point was a prostate-specific antigen (PSA) decline of > or = 50%. Secondary end points included time to progression, overall survival, and safety. Results In the mouse model, combination therapy of docetaxel, bevacizumab, and thalidomide inhibited tumor growth most effectively. In the clinical trial, 90% of patients receiving the combination therapy had PSA declines of > or = 50%, and 88% achieved a PSA decline of > or = 30% within the first 3 months of treatment. The median time to progression was 18.3 months, and the median overall survival was 28.2 months in this group with a Halabi-predicted survival of 14 months. While toxicities were manageable, all patients developed grade 3/4 neutropenia. CONCLUSION The addition of bevacizumab and thalidomide to docetaxel is a highly active combination with manageable toxicities. The estimated median survival is encouraging, given the generally poor prognosis of this patient population. These results suggest that definitive clinical trials combining antiangiogenic agent combinations with docetaxel are warranted to improve treatment outcomes for patients with metastatic CRPC.
• Femoral-based Central Venous Oxygen Saturation is Not a Reliable Substitute for Subclavian/internal Jugular-based Central Venous Oxygen Saturation in Patients Who Are Critically Ill Chest. Jul, 2010  |   Pubmed ID: 20418366 Central venous oxygen saturation (Scv(O(2))) has been used as a surrogate marker for mixed venous oxygen saturation (Sv(O(2))). Femoral venous oxygen saturation (Sfv(O(2))) is sometimes used as a substitute for Scv(O(2)). The purpose of this study is to test the hypothesis that these values can be used interchangeably in a population of patients who are critically ill.
• Metal Exposures in an Inner-city Neonatal Population Environment International. Oct, 2010  |   Pubmed ID: 20553999 We measured concentrations of lead (Pb), manganese (Mn), chromium (Cr), and copper (Cu) in umbilical cord whole blood and examined sources of environmental Pb exposures in a predominantly African-American population.
• Suicide Prevention Through Stories of Hope Journal of Christian Nursing : a Quarterly Publication of Nurses Christian Fellowship. Jul-Sep, 2010  |   Pubmed ID: 20632483 A challenge in preventing suicide is to provide a person hope so that she or he can see that suicide is not the only way out. Narratives or stories from varied sources can be a helpful tool to assist patients in putting a confusing or frightening reality into some sort of framework. Whatever one's theological preference, biblical narratives can place untenable events into a positive, hopeful mental framework, helping people make structure and sense out of seeming chaos.
• An Efficient Route to Xanthine Based A(2A) Adenosine Receptor Antagonists and Functional Derivatives Organic & Biomolecular Chemistry. Sep, 2010  |   Pubmed ID: 20652178 A one-pot route to 8-substituted xanthines has been developed from 5,6-diaminouracils and carboxaldehydes. Yields are good and the process applicable to a range of substrates including a family of A(2A) adenosine receptor antagonists. A new route to the KW-6002 family of antagonists is presented including a pro-drug variant, and application to related image contrast agents developed.
• The Lysophosphatidic Acid (LPA) and Sphingosine-1-phosphate (S1P) Receptor Gene Families: Cloning and Comparative Expression Analysis in Xenopus Laevis The International Journal of Developmental Biology. 2010  |   Pubmed ID: 20712001 Sphingosine-1-phosphate (S1P) and lysophosphatidic acid (LPA) are endogenous bioactive lipids which mediate a variety of biological cell responses such as cell proliferation, migration, differentiation and apoptosis. Their actions are mediated by binding to the G-protein-coupled endothelial differentiation gene (Edg) receptor subfamily, referred to as S1P1-5 and LPA1-5, and regulate a variety of signalling pathways involved in numerous physiological processes and pathological conditions. Their importance during embryogenesis has been demonstrated by the generation of knock-out mice and specific roles have been assigned to these receptors. However, potential functional redundancy and the lethality of some mutants have complicated functional analysis in these models. Here we report the cloning of the S1P and LPA receptors in Xenopus laevis and tropicalis. Phylogenetic analyses demonstrate the high level of conservation of these receptors between amphibian and other vertebrate species. We have conducted a comparative expression analysis of these receptors during development and in the adult frog, by both RT-PCR and whole mount in situ hybridisation. In particular, we show that S1P1, 2 and 5 display distinct embryonic specific expression patterns, suggesting potentially different developmental roles for these receptors, and therefore for their ligands, during amphibian embryogenesis.
• Stimulation of Methane Generation from Nonproductive Coal by Addition of Nutrients or a Microbial Consortium Applied and Environmental Microbiology. Nov, 2010  |   Pubmed ID: 20817801 Biogenic formation of methane from coal is of great interest as an underexploited source of clean energy. The goal of some coal bed producers is to extend coal bed methane productivity and to utilize hydrocarbon wastes such as coal slurry to generate new methane. However, the process and factors controlling the process, and thus ways to stimulate it, are poorly understood. Subbituminous coal from a nonproductive well in south Texas was stimulated to produce methane in microcosms when the native population was supplemented with nutrients (biostimulation) or when nutrients and a consortium of bacteria and methanogens enriched from wetland sediment were added (bioaugmentation). The native population enriched by nutrient addition included Pseudomonas spp., Veillonellaceae, and Methanosarcina barkeri. The bioaugmented microcosm generated methane more rapidly and to a higher concentration than the biostimulated microcosm. Dissolved organics, including long-chain fatty acids, single-ring aromatics, and long-chain alkanes accumulated in the first 39 days of the bioaugmented microcosm and were then degraded, accompanied by generation of methane. The bioaugmented microcosm was dominated by Geobacter sp., and most of the methane generation was associated with growth of Methanosaeta concilii. The ability of the bioaugmentation culture to produce methane from coal intermediates was confirmed in incubations of culture with representative organic compounds. This study indicates that methane production could be stimulated at the nonproductive field site and that low microbial biomass may be limiting in situ methane generation. In addition, the microcosm study suggests that the pathway for generating methane from coal involves complex microbial partnerships.
• Orthopedic Trauma-induced Pulmonary Injury in the Obese Zucker Rat Microcirculation (New York, N.Y. : 1994). Nov, 2010  |   Pubmed ID: 21044219 Obese subjects with orthopedic trauma exhibit increased inflammation and an increased risk of pulmonary edema. Prostaglandin E(2) (PGE(2) ) production is elevated during inflammation and associated with increased vascular permeability. We hypothesize that pulmonary edema in obesity following orthopedic trauma is due to elevated PGE(2) and resultant increases in pulmonary permeability.
• SNM Practice Guideline for Breast Scintigraphy with Breast-specific Gamma-cameras 1.0 Journal of Nuclear Medicine Technology. Dec, 2010  |   Pubmed ID: 21057112
• Is the Drip-and-ship Approach to Delivering Thrombolysis for Acute Ischemic Stroke Safe? The Journal of Emergency Medicine. Aug, 2011  |   Pubmed ID: 19272734 The drip-and-ship method of treating stroke patients may increase the use of tissue plasminogen activator (t-PA) in community hospitals.
• Zero on the NIHSS Does Not Equal the Absence of Stroke Annals of Emergency Medicine. Jan, 2011  |   Pubmed ID: 20828876 The National Institutes of Health Stroke Scale (NIHSS) measures deficits caused by a stroke, but not all stroke signs are captured on the NIHSS. We determine the symptoms and stroke localization of patients with brain infarction and an NIHSS score of 0.
• Basal Cell Carcinomas in Gorlin Syndrome: a Review of 202 Patients Journal of Skin Cancer. 2011  |   Pubmed ID: 21152126 Gorlin syndrome (Naevoid Basal Cell Carcinoma Syndrome) is a rare autosomal dominant syndrome caused by mutations in the PTCH gene with a birth incidence of approximately 1 in 19,000. Patients develop multiple basal cell carcinomas of the skin frequently in early life and also have a predisposition to additional malignancies such as medulloblastoma. Gorlin Syndrome patients also have developmental defects such as bifid ribs and other complications such as jaw keratocysts. We studied the incidence and frequency of basal cell carcinomas in 202 Gorlin syndrome patients from 62 families and compared this to their gender and mutation type. Our data suggests that the incidence of basal cell carcinomas is equal between males and females and the mutation type cannot be used to predict disease burden.
• Asymmetric Synthesis of α-aryl Amino Acids; Aryne-mediated Diastereoselective Arylation Organic Letters. Mar, 2011  |   Pubmed ID: 21302900 An aryne-mediated α-arylation reaction of Schöllkopf's bis-lactim ether is described. Arynes were generated via an ortho-lithiation approach, affording syn-arylated products in up to 94:6 dr with moderate to good yields and excellent regioselectivities. Hydrolysis provided a variety of substituted arylglycines containing a range of functional groups without racemization.
• Recognizing Hospital-acquired Burn Injury in Patients After Coronary Artery Bypass Surgery Journal of Wound, Ostomy, and Continence Nursing : Official Publication of The Wound, Ostomy and Continence Nurses Society / WOCN. Mar-Apr, 2011  |   Pubmed ID: 21389831 Nosocomial injury is a constant threat in the hospital setting. While there is growing awareness surrounding hospital-acquired pressure ulcers, little information is available on burns associated with intraoperative procedures.
• Mechanical Factors in the Development of the Vascular Bed Respiratory Physiology & Neurobiology. Aug, 2011  |   Pubmed ID: 21458600 During embryonic development, blood flow is needed not only to nourish the developing embryo but is also important for shaping the vascular network such that it becomes hemodynamically efficient. The first blood vessels form a network called the capillary plexus. After the onset of blood flow, the capillary plexus remodel into a more hierarchical tree-shaped network. Mechanical forces created by blood flow are required for remodelling to occur and these forces are believed to induce a maturation of the blood vessels that stabilizes the growing vascular network. The role of mechanical force has been extensively studied in the mature cardiovascular system. Though the events induced by blood flow during development are thought to be similar to what occurs in the adult, there are several important differences between the embryo and the adult. We therefore discuss what is known about the role of mechanical forces in vascular remodelling from the adult cardiovascular system and highlight how embryonic development differs from the adult. We consider the role of blood flow in altering branching morphology, arterial-venous identity and the formation of the blood vessel wall during early vascular development.
• Pronephric Tubulogenesis Requires Daam1-mediated Planar Cell Polarity Signaling Journal of the American Society of Nephrology : JASN. Sep, 2011  |   Pubmed ID: 21804089 Canonical β-catenin-mediated Wnt signaling is essential for the induction of nephron development. Noncanonical Wnt/planar cell polarity (PCP) pathways contribute to processes such as cell polarization and cytoskeletal modulation in several tissues. Although PCP components likely establish the plane of polarization in kidney tubulogenesis, whether PCP effectors directly modulate the actin cytoskeleton in tubulogenesis is unknown. Here, we investigated the roles of Wnt PCP components in cytoskeletal assembly during kidney tubule morphogenesis in Xenopus laevis and zebrafish. We found that during tubulogenesis, the developing pronephric anlagen expresses Daam1 and its interacting Rho-GEF (WGEF), which compose one PCP/noncanonical Wnt pathway branch. Knockdown of Daam1 resulted in reduced expression of late pronephric epithelial markers with no apparent effect upon early markers of patterning and determination. Inhibiting various points in the Daam1 signaling pathway significantly reduced pronephric tubulogenesis. These data indicate that pronephric tubulogenesis requires the Daam1/WGEF/Rho PCP pathway.
• Wilms Tumor Incidence in Children with 2q Terminal Deletions: a Cohort Study American Journal of Medical Genetics. Part A. Sep, 2011  |   Pubmed ID: 21815249 Three individuals with chromosome 2q terminal deletions have been reported in the medical literature to have developed Wilms tumor. By looking at a UK national cohort, we aimed to ascertain the chance of an individual with a 2q terminal deletion developing a Wilms tumor. The objective was to clarify screening recommendations. All individuals over a 40-year period with chromosome 2q terminal deletions were ascertained from the Chromosome Abnormality Database. The names and dates of birth of these individuals were obtained from the Regional Cytogenetic Departments where the original chromosome analyses were performed. These data were collated and compared with the National Registry of Childhood Tumors. One hundred twenty-nine subjects were identified over a 40-year study period. Only a single individual in our national cohort was affected by Wilms tumor. This individual had an add(2)(q35) karyotype. We conclude that the incidence of Wilms tumor in the majority of individuals with a 2q terminal deletion is low, and is below the recommended threshold for surveillance for tumor development.
• Rheology of Embryonic Avian Blood American Journal of Physiology. Heart and Circulatory Physiology. Dec, 2011  |   Pubmed ID: 21963831 Shear stress, a mechanical force created by blood flow, is known to affect the developing cardiovascular system. Shear stress is a function of both shear rate and viscosity. While established techniques for measuring shear rate in embryos have been developed, the viscosity of embryonic blood has never been known but always assumed to be like adult blood. Blood is a non-Newtonian fluid, where the relationship between shear rate and shear stress is nonlinear. In this work, we analyzed the non-Newtonian behavior of embryonic chicken blood using a microviscometer and present the apparent viscosity at different hematocrits, different shear rates, and at different stages during development from 4 days (Hamburger-Hamilton stage 22) to 8 days (about Hamburger-Hamilton stage 34) of incubation. We chose the chicken embryo since it has become a common animal model for studying hemodynamics in the developing cardiovascular system. We found that the hematocrit increases with the stage of development. The viscosity of embryonic avian blood in all developmental stages studied was shear rate dependent and behaved in a non-Newtonian manner similar to that of adult blood. The range of shear rates and hematocrits at which non-Newtonian behavior was observed is, however, outside the physiological range for the larger vessels of the embryo. Under low shear stress conditions, the spherical nucleated blood cells that make up embryonic blood formed into small aggregates of cells. We found that the apparent blood viscosity decreases at a given hematocrit during embryonic development, not due to changes in protein composition of the plasma but possibly due to the changes in cellular composition of embryonic blood. This decrease in apparent viscosity was only visible at high hematocrit. At physiological values of hematocrit, embryonic blood viscosity did not change significantly with the stage of development.
• The Initiation of Blood Flow and Flow Induced Events in Early Vascular Development Seminars in Cell & Developmental Biology. Dec, 2011  |   Pubmed ID: 22001248 Within a day of gastrulation, the embryonic heart begins to beat and creates blood flow in the developing cardiovascular system. The onset of blood flow completely changes the environment in which the cardiovascular system is forming. Flow provides physiological feedback such that the developing network adapts to cue provided by the flow. Targeted inactivation of genes that alter early blood fluid dynamics induce secondary defects in the heart and vasculature and therefore proper blood flow is known to be essential for vascular development. Though hemodynamics, or blood fluid dynamics, are known to activate signaling pathways in the mature cardiovascular system in pathologies ranging from artherosclerosis to angiogenesis, the role in development has not been as intensively studied. The question arises how blood vessels in the embryos, which initially lack cells types such as smooth muscle cells, differ in their response to mechanical signals from blood flow as compared to the more mature cardiovascular system. Many genes known to be regulated by hemodynamics in the adult are important for developmental angiogenesis. Therefore the onset of blood flow is of primary importance to vascular development. This review will focus on how blood flow initiates and the effects of the mechanical signals created by blood flow on cardiovascular development.
• A Pilot Study of MUC-1/CEA/TRICOM Poxviral-based Vaccine in Patients with Metastatic Breast and Ovarian Cancer Clinical Cancer Research : an Official Journal of the American Association for Cancer Research. Nov, 2011  |   Pubmed ID: 22068656 PANVAC is a recombinant poxviral vaccine that contains transgenes for MUC-1, CEA, and 3 T-cell costimulatory molecules. This study was conducted to obtain preliminary evidence of clinical response in metastatic breast and ovarian cancer patients.
• Periodic Limb Movements During Sleep in Children with Narcolepsy Journal of Clinical Sleep Medicine : JCSM : Official Publication of the American Academy of Sleep Medicine. Dec, 2011  |   Pubmed ID: 22171197 In adults with narcolepsy, periodic limb movements of sleep (PLMS) occur more frequently than in control population, and presence of increased PLMS is associated with greater sleep disruption and shorter mean sleep latency. This study was performed to determine whether PLMS are common in children with narcolepsy, and whether the presence of PLMS is associated with greater sleep disruption.
• Asymmetric Synthesis of Quaternary Aryl Amino Acid Derivatives Via a Three-component Aryne Coupling Reaction Beilstein Journal of Organic Chemistry. 2011  |   Pubmed ID: 22238534 A method was developed for the synthesis of α-alkyl, α-aryl-bislactim ethers in good to excellent yields and high diastereoselectivities, consisting of a facile one-pot procedure in which the aryl group is introduced by means of a nucleophilic addition to benzyne and the alkyl group by alkylation of a resultant benzylic anion. Hydrolysis of the sterically less hindered adducts gave the corresponding quaternary amino acids with no racemization, whereas hydrolytic ring opening gave the corresponding valine dipeptides from bulkier bislactims.
• Family Hardship, Family Instability, and Cognitive Development Journal of Epidemiology and Community Health. Aug, 2012  |   Pubmed ID: 21507894 Associations between the characteristics of the family environment, in particular poverty and family structure, and cognitive development are well established, yet little is known about the role of timing and accumulation of risk in early childhood. The aim of this paper is to assess the associations between income poverty, family instability and cognitive development in early childhood. In particular, it tests the relative role of family economic hardship compared with family instability in affecting cognitive functioning at the age of 5 years.
• The Prevalence and Management of Low Back Pain Across Adulthood: Results from a Population-based Cross-sectional Study (the MUSICIAN Study) Pain. Jan, 2012  |   Pubmed ID: 21978663 The aim of the current study was to determine: the prevalence of low back pain (LBP) and associated disability; the frequency of consultation to general practice; whether there were differences in management by age. We conducted a cross-sectional population study in Aberdeen city and Cheshire County, UK. Participants were 15,272 persons aged 25 years and older. The 1-month period prevalence of LBP was 28.5%. It peaked at age 41-50 years, but at ages over 80 years was reported by 1 in 4 persons. Older persons were more likely to consult, and the prevalence of severe LBP continued to increase with age. Management by general practitioners differed by age of the patient. Older persons (> 70 vs ≤ 40 years) were more likely to only have been prescribed painkillers (odds ratio [OR] 1.74, 95% confidence interval [CI] 1.28-2.35) or only pain killers with other medications (OR 1.45, 95% CI 1.07-1.98). They were less likely to be prescribed physiotherapy or exercise (OR 0.63, 95% CI 0.46-0.85) or to be referred to a specialist (OR 0.77, 95% CI 0.57-1.04). Older persons were more likely to have previously received exercise therapy for pain, were less likely to be enthusiastic about receiving it now (P
• Curative Treatment Without Surgical Reconstruction After Perineal Debridement of Fournier's Gangrene Journal of Wound, Ostomy, and Continence Nursing : Official Publication of The Wound, Ostomy and Continence Nurses Society / WOCN. Jan-Feb, 2012  |   Pubmed ID: 22237647 Fournier's gangrene (necrotizing fasciitis) is an acute life-threatening disease of the perineal area that requires urgent medical intervention. Once the affected area is surgically debrided and the patient is stabilized, surgical management typically involves 1 or more additional procedures that may include split-thickness skin grafts, flaps, or an elective diverting urostomy and/or colostomy. The professional literature discussing nonsurgical approaches to healing for Fournier's gangrene after surgical debridement is sparse.
• Comparison of Trihalomethanes in Tap Water and Blood: a Case Study in the United States Environmental Health Perspectives. May, 2012  |   Pubmed ID: 22281753 Epidemiological studies have used various measures to characterize trihalomethane (THM) exposures, but the relationship of these indicators to exposure biomarkers remains unclear.
• Assessing Hepatomegaly: Automated Volumetric Analysis of the Liver Academic Radiology. May, 2012  |   Pubmed ID: 22361033 The aims of this study were to define volumetric nomograms for identifying hepatomegaly and to retrospectively evaluate the performance of radiologists in assessing hepatomegaly.
• Efficient Synthesis of Sterically-stabilized Nano-objects Via RAFT Dispersion Polymerization of Benzyl Methacrylate in Alcoholic Media Advanced Materials (Deerfield Beach, Fla.). Jul, 2012  |   Pubmed ID: 22605479 Synthesis of diblock copolymer nano-objects: alcohol is a good idea! RAFT dispersion polymerization of benzyl methacrylate in alcohol using weak polyelectrolyte-based chain transfer agents allows the facile synthesis of sterically stabilized diblock copolymer nano-objects with very high monomer conversions. Such syntheses are usually problematic when conducted in water due to electrostatic repulsion between highly charged stabilizer chains, which impedes in situ self-assembly. Construction of a detailed phase diagram facilitates reproducible syntheses of well-defined diblock copolymer spheres, worms or vesicles, since it allows mixed phase regions to be avoided. Aqueous electrophoresis studies confirm that these nano-objects can acquire substantial surface charge when transferred to aqueous solution due to ionization (or protonation) of the polyacid (or polybase) stabilizer chains.
• The JAMA Network Website: Today's Content on the Future of Medical Publishing JAMA : the Journal of the American Medical Association. Jun, 2012  |   Pubmed ID: 22706839
• Diagnosing Fetal Alcohol Syndrome: New Insights from Newer Genetic Technologies Archives of Disease in Childhood. Sep, 2012  |   Pubmed ID: 22798695 A genetic opinion is frequently requested in the assessment of a child with suspected fetal alcohol spectrum disorders (FASD). We studied the outcome of genetic assessment of 80 children referred to a regional genetics centre between 2004 and 2010 to identify the value of the genetic assessment in cases of suspected FASD.
• Time-lapse Microscopy of Macrophages During Embryonic Vascular Development Developmental Dynamics : an Official Publication of the American Association of Anatomists. Sep, 2012  |   Pubmed ID: 22815139 Macrophages are present before the onset of blood flow, but very little is known about their function in vascular development. We have developed a technique to concurrently label both endothelial cells and macrophages for time-lapse microscopy using co-injection of fluorescently conjugated acetylated low-density lipoprotein (AcLDL) and phagocytic dye PKH26-PCL.
• The Glycocalyx is Present As Soon As Blood Flow is Initiated and is Required for Normal Vascular Development Developmental Biology. Sep, 2012  |   Pubmed ID: 22820069 The glycocalyx, and the thicker endothelial surface layer (ESL), are necessary both for endothelial barrier function and for sensing mechanical forces in the adult. The goal of this study is to use a combination of imaging techniques to establish when the glycocalyx and endothelial surface layer form during embryonic development and to determine the biological significance of the glycocalyx layer during vascular development in quail embryos. Using transmission electron microscopy, we show that the glycocalyx layer is present as soon as blood flow starts (14 somites). The early endothelial glycocalyx (14 somites) lacks the distinct hair-like morphology that is present later in development (17 and 25 somites). The average thickness does not change significantly (14 somites, 182 nm ± 33 nm; 17 somites, 218 ± 30 nm; 25 somites, 212 ± 32 nm). The trapping of circulating fluorescent albumin was used to evaluate the development of the ESL. Trapped fluorescent albumin was first observed at 25 somites. In order to assess a functional role for the glycocalyx during development, we selectively degraded luminal glycosaminoglycans. Degradation of hyaluronan compromised endothelial barrier function and prevented vascular remodeling. Degradation of heparan sulfate down regulated the expression of shear-sensitive genes but does not inhibit vascular remodeling. Our findings show that the glycocalyx layer is present as soon as blood flow starts (14 somites). Selective degradations of major glycocalyx components were shown to inhibit normal vascular development, examined through morphology, vascular barrier function, and gene expression.
• Use of Inert Gas Jets to Measure the Forces Required for Mechanical Gene Transfection Biomedical Engineering Online. 2012  |   Pubmed ID: 22963645 Transferring genes and drugs into cells is central to how we now study, identify and treat diseases. Several non-viral gene therapy methods that rely on the mechanical disruption of the plasma membrane have been proposed, but the success of these methods has been limited due to a lack of understanding of the mechanical parameters that lead to cell membrane permeability.
• National Recruitment in Dentistry: the Orthodontic Experience Journal of Orthodontics. Sep, 2012  |   Pubmed ID: 22984097
• Increased Shear Stress Inhibits Angiogenesis in Veins and Not Arteries During Vascular Development Angiogenesis. Jan, 2013  |   Pubmed ID: 22941228 Vascular development is believed to occur first by vasculogenesis followed by angiogenesis. Though angiogenesis is the formation of new vessels, we found that vascular density actually decreases during this second stage. The onset of the decrease coincided with the entry of erythroblasts into circulation. We therefore measured the level of shear stress at various developmental stages and found that it was inversely proportional to vascular density. To investigate whether shear stress was inhibitory to angiogenesis, we altered shear stress levels either by preventing erythroblasts from entering circulation ("low" shear stress) or by injection of a starch solution to increase the blood plasma viscosity ("high" shear stress). By time-lapse microscopy, we show that reverse intussusception (merging of two vessels) is inversely proportional to the level of shear stress. We also found that angiogenesis (both sprouting and splitting) was inversely proportional to shear stress levels. These effects were specific to the arterial or venous plexus however, such that the effect on reverse intussusception was present only in the arterial plexus and the effect on sprouting only in the venous plexus. We cultured embryos under altered shear stress in the presence of either DAPT, a Notch inhibitor, or DMH1, an inhibitor of the bone morphogenetic protein (BMP) pathway. DAPT treatment phenocopied the inhibition of erythroblast circulation ("low" shear stress) and the effect of DAPT treatment could be partially rescued by injection of starch. Inhibition of the BMP signaling prevented the reduction in vascular density that was observed when starch was injected to increase shear stress levels.
• LCL124, a Cationic Analog of Ceramide, Selectively Induces Pancreatic Cancer Cell Death by Accumulating in Mitochondria The Journal of Pharmacology and Experimental Therapeutics. Jan, 2013  |   Pubmed ID: 23086228 Treatment of pancreatic cancer that cannot be surgically resected currently relies on minimally beneficial cytotoxic chemotherapy with gemcitabine. As the fourth leading cause of cancer-related death in the United States with dismal survival statistics, pancreatic cancer demands new and more effective treatment approaches. Resistance to gemcitabine is nearly universal and appears to involve defects in the intrinsic/mitochondrial apoptotic pathway. The bioactive sphingolipid ceramide is a critical mediator of apoptosis initiated by a number of therapeutic modalities. It is noteworthy that insufficient ceramide accumulation has been linked to gemcitabine resistance in multiple cancer types, including pancreatic cancer. Taking advantage of the fact that cancer cells frequently have more negatively charged mitochondria, we investigated a means to circumvent resistance to gemcitabine by targeting delivery of a cationic ceramide (l-t-C6-CCPS [LCL124: ((2S,3S,4E)-2-N-[6'-(1″-pyridinium)-hexanoyl-sphingosine bromide)]) to cancer cell mitochondria. LCL124 was effective in initiating apoptosis by causing mitochondrial depolarization in pancreatic cancer cells but demonstrated significantly less activity against nonmalignant pancreatic ductal epithelial cells. Furthermore, we demonstrate that the mitochondrial membrane potentials of the cancer cells were more negative than nonmalignant cells and that dissipation of this potential abrogated cell killing by LCL124, establishing that the effectiveness of this compound is potential-dependent. LCL124 selectively accumulated in and inhibited the growth of xenografts in vivo, confirming the tumor selectivity and therapeutic potential of cationic ceramides in pancreatic cancer. It is noteworthy that gemcitabine-resistant pancreatic cancer cells became more sensitive to subsequent treatment with LCL124, suggesting that this compound may be a uniquely suited to overcome gemcitabine resistance in pancreatic cancer.
• The 60-day Temperature-dependent Degradation of Midazolam and Lorazepam in the Prehospital Environment Prehospital Emergency Care : Official Journal of the National Association of EMS Physicians and the National Association of State EMS Directors. Jan-Mar, 2013  |   Pubmed ID: 23148574 The choice of the optimal benzodiazepine to treat prehospital status epilepticus is unclear. Lorazepam is preferred in the emergency department, but concerns about nonrefrigerated storage limits emergency medical services (EMS) use. Midazolam is increasingly popular, but its heat stability is undocumented.
• Health Maintenance in Women American Family Physician. Jan, 2013  |   Pubmed ID: 23317023 The health maintenance examination is an opportunity to focus on disease prevention and health promotion. The patient history should include screening for tobacco use, alcohol misuse, intimate partner violence, and depression. Premenopausal women should receive preconception counseling and contraception as needed, and all women planning or capable of pregnancy should take 400 to 800 mcg of folic acid per day. High-risk sexually active women should be counseled on reducing the risk of sexually transmitted infections, and screened for chlamydia, gonorrhea, and syphilis. All women should be screened for human immunodeficiency virus. Adults should be screened for obesity and elevated blood pressure. Women 20 years and older should be screened for dyslipidemia if they are at increased risk of coronary heart disease. Those with sustained blood pressure greater than 135/80 mm Hg should be screened for type 2 diabetes mellitus. Women 55 to 79 years of age should take 75 mg of aspirin per day when the benefits of stroke reduction outweigh the increased risk of gastrointestinal hemorrhage. Women should begin cervical cancer screening by Papanicolaou test at 21 years of age, and if results have been normal, screening may be discontinued at 65 years of age or after total hysterectomy. Breast cancer screening with mammography may be considered in women 40 to 49 years of age based on patients' values, and potential benefits and harms. Mammography is recommended biennially in women 50 to 74 years of age. Women should be screened for colorectal cancer from 50 to 75 years of age. Osteoporosis screening is recommended in women 65 years and older, and in younger women with a similar risk of fracture. Adults should be immunized at recommended intervals according to guidelines from the Centers for Disease Control and Prevention.
• Cardiorenal Anemia Syndrome As a Prognosticator for Death in Heart Failure The American Journal of Cardiology. Apr, 2013  |   Pubmed ID: 23375730 Anemia and chronic kidney disease are common in patients with heart failure (HF) and are associated with adverse outcomes. We analyzed the effect of cardiorenal anemia (CRA) syndrome, defined as anemia (hemoglobin
• A Prospective Clinical Study to Evaluate Early Success of Short Implants The International Journal of Oral & Maxillofacial Implants. Jan-Feb, 2013  |   Pubmed ID: 23377063 Short implants are an alternative to bone augmentation procedures for patients with reduced bone height. This study evaluated the success of short implants in posterior locations prior to loading.
• Automated Detection of Sclerotic Metastases in the Thoracolumbar Spine at CT Radiology. Jul, 2013  |   Pubmed ID: 23449957 To design and validate a computer system for automated detection and quantitative characterization of sclerotic metastases of the thoracolumbar spine on computed tomography (CT) images.
• Early Age of First Sex and Health Risk in an Urban Adolescent Population The Journal of School Health. May, 2013  |   Pubmed ID: 23517003 Early sex is associated with high-risk behaviors and outcomes, including sexual risk behaviors, forced sex, physical dating violence, and becoming pregnant or impregnating someone.
• Assessing Splenomegaly: Automated Volumetric Analysis of the Spleen Academic Radiology. Jun, 2013  |   Pubmed ID: 23535191 To define systematic volumetric thresholds to identify and grade splenomegaly and retrospectively evaluate the performance of radiologists to assess splenomegaly in computed tomography (CT) image data.
• Risky Removal: Developing a Holistic Understanding of the Risks of Redeveloping Sites Contaminated with Unexploded Ordnance Environmental Science & Technology. May, 2013  |   Pubmed ID: 23607512
• Preterm Birth: Strategies for Establishing Adequate Milk Production and Successful Lactation Seminars in Fetal & Neonatal Medicine. Apr, 2013  |   Pubmed ID: 23623976 Whilst human milk has not evolved to meet the unique requirements of the preterm infant there are unquestionable benefits to be gained via breast milk in terms of the development and health of the infant. Many mothers of preterm infants struggle to achieve a full milk production for many reasons the mechanisms of which are still unclear. Strategies to enhance milk volume include early, frequent simultaneous expression of milk combined with breast massage and a reduction of stress. However, these are not always successful, therefore a greater understanding of lactation physiology is required to devise more effective interventions to increase milk supply. The difficulty these infants experience transitioning to oral feeding and ultimately full breastfeeding further complicates lactation. In order to improve the health of these already compromised infants it is critical that more research be directed to this area so that they reap all the benefits that can be gained from breastfeeding.
• Embryogenesis of the First Circulating Endothelial Cells PloS One. 2013  |   Pubmed ID: 23737938 Prior to this study, the earliest appearance of circulating endothelial cells in warm-blooded animals was unknown. Time-lapse imaging of germ-line transformed Tie1-YFP reporter quail embryos combined with the endothelial marker antibody QH1 provides definitive evidence for the existence of circulating endothelial cells - from the very beginning of blood flow. Blood-smear counts of circulating cells from Tie1-YFP embryos showed that up to 30% of blood-borne cells are Tie1 positive; though cells expressing low levels of YFP were also positive for benzidine, a hemoglobin stain, suggesting that these cells were differentiating into erythroblasts. Electroporation-based time-lapse experiments, exclusively targeting the intra-embryonic mesoderm were combined with QH1 immunostaining. The latter antibody marks quail endothelial cells. Together the optical data provide conclusive evidence that endothelial cells can enter blood flow from vessels of the embryo proper, as well as from extra-embryonic areas. When Tie1-YFP positive cells and tissues are transplanted to wild type host embryos, fluorescent cells emigrate from such transplants and join host vessels; subsequently a few YFP cells are shed into circulation. These data establish that entering circulation is a commonplace activity of embryonic vascular endothelial cells. We conclude that in the class of vertebrates most closely related to mammals a normal component of primary vasculogenesis is production of endothelial cells that enter circulation from all vessels, both intra- and extra-embryonic.
• Prevalence of Obesity and Abdominal Obesity from Four to 16 Years Old Children Living in the Mexico-USA Border Nutrición Hospitalaria. Mar-Apr, 2013  |   Pubmed ID: 23822701 The prevalence of obesity among Mexicans is alarming in both the child and adult populations. The objective of this study was to determine the levels of overweight, obesity and abdominal obesity in pre-school (PS), elementary (ES), and middle high (MHS) public school children from Tijuana. From February to April of 2011, a bietapic random sample was selected by cluster method of 30 PS, 30 ES, and 30 MHS children. And a sample of 30 groups for each level was chosen. Twenty elementary teachers and eight graduate students were trained at one central location on how to take anthropometric measurements using a portable scale, a stadiometer, and a measuring tape to determine weight, height, and waist circumference. Body Mass Index values were computed and compared to age/gender BMI percentiles according to WHO criteria. Waist circumference for-age at the 90th percentile from NHANES III (Mexican-American) was used to define abdominal obesity. The sample was composed of 646 PS children, 961 ES children, and 1,095 MHS children. Their ages ranged from 4- 16 years. Results showed an overall prevalence of overweight and obesity in younger than 5y preschool children (> 2 SD) of 23.1%, in ≥ 5 y PS (> 1 SD) of 33.8%, in ES children of 46.3%, and in MHS children of 41.9%. Abdominal obesity in PS children was 18%, in ES children was 16.7%, and in MHS children was 15.2%. These results warrant immediate and comprehensive actions to prevent a critical public health problem in Mexico.