Toward Increased Utility of MtDNA in Forensic Identifications

Forensic Science International. Dec, 2004  |  Pubmed ID: 15639563

The utility of mtDNA in forensic identifications is limited by its low power of discrimination and the absence of high quality mtDNA databases. Single nucleotide polymorphisms (SNPs) in the control region outside of hypervariable regions I and II (HVI/HVII), and in the coding region of the mtDNA genome, can provide additional discrimination in mtDNA testing. We have identified particularly useful SNP sites via high throughput sequencing of the entire mtDNA genome. We report here two cases in which an 11-plex SNP assay (panel "A") targeting the most common HVI/HVII type successfully resolved two cases in which identifications could not be made on the basis of HVI/HVII sequencing. Additionally, we established a database of 286 samples for SNP panel "A" generated with robotic protocols. We have addressed the need for high quality mtDNA control region (CR) databases by developing robotic protocols for lab processing, and a carefully devised electronic data review process. A large-scale databasing effort targeting several populations underrepresented in current mtDNA databases is underway.

The Structure of Carbonmonoxy Neuroglobin Reveals a Heme-sliding Mechanism for Control of Ligand Affinity

Proceedings of the National Academy of Sciences of the United States of America. Dec, 2004  |  Pubmed ID: 15548613

Neuroglobin (Ngb), a globular heme protein expressed in the brain of vertebrates, binds oxygen reversibly, with an affinity comparable to myoglobin (Mb). Despite low sequence identity, the overall 3D fold of Ngb and Mb is very similar. Unlike in Mb, in Ngb the sixth coordination position of the heme iron is occupied by the distal histidine, in the absence of an exogenous ligand. Endogenous ligation has been proposed as a unique mechanism for affinity regulation and ligand discrimination in heme proteins. This peculiarity might be related to the still-unknown physiological function of Ngb. Here, we present the x-ray structure of CO-bound ferrous murine Ngb at 1.7 A and a comparison with the 1.5-A structure of ferric bis-histidine Ngb. We have also used Fourier transform IR spectroscopy of WT and mutant CO-ligated Ngb to examine structural heterogeneity in the active site. Upon CO binding, the distal histidine retains (by and large) its position, whereas the heme group slides deeper into a preformed crevice, thereby reshaping the large cavity ( approximately 290 A(3)) connecting the distal and proximal heme sides with the bulk. The heme relocation is accompanied by a significant decrease of structural disorder, especially of the EF loop, which may be the signal whereby Ngb communicates hypoxic conditions. This unexpected structural change unveils a heme-sliding mechanism of affinity control that may be of significance to understanding Ngb's role in the pathophysiology of the brain.

Genotyping SNPs Using a UV-photocleavable Oligonucleotide in MALDI-TOF MS

Methods in Molecular Biology (Clifton, N.J.). 2005  |  Pubmed ID: 15570107

Matrix-assisted laser desorption time-of-flight mass spectrometry coupled with allele-specific primer extension is a proven method for typing single nucleotide polymorphisms (SNPs). A novel modification upon this methodology is the incorporation of a photocleavable linker within the extension primer. After completion of the primer extension reaction, photocleavage of the extension products results in two deoxyribonucleic acid (DNA) fragments of lower mass. Typically, the smaller cleavage product, which contains the genotyping information, is in the range of 1000-3000 Daltons. The decrease in primer mass allows for higher sensitivity in mass spectrometric measurement and increases the potential for higher levels of multiplexing. The disturbing mass spectrometric analysis peaks caused by salt adducts and doubly charged ions are diminished when analyzing lower-mass DNA fragments. Here, we illustrate the methodology for using photocleavable modified extension primers for detection of SNPs located on the Y chromosome. Genomic templates were prepared from anonymous male donors. Five regions of the Y chromosome containing the SNP markers M9, M42, M45, M89, and M96 were amplified by polymerase chain reaction, treated with shrimp alkaline phosphatase, and subjected to primer extension reactions using primers containing a photocleavable building block at specific sites. After elongation, the extension primers were desalted and subjected to ultraviolet irradiation to cleave the products at the photocleavable site. Subsequently, the small fragments derived from the 3' ends of the molecules containing the genotype information were analyzed by matrix-assisted laser desorption ionization time-of-flight using a 3-hydroxypicolinic acid matrix.

The Acromegalic Arthropathy

Journal of Endocrinological Investigation. 2005  |  Pubmed ID: 16323826

Since the initial definition of acromegaly over 100 yr ago, extraordinary efforts have been made to control GH levels and the pituitary tumor. Criteria of cure have been evolving constantly and all previous statements should be revised according to modern criteria of disease control. Arthropathy is undoubtedly the most important cause of morbidity and functional disability of acromegaly, and bone alterations are highly characteristic of this syndrome. They involve theoretically all bones and, particularly, the appendicular and the axial skeleton. Radiographic changes at different joint sites are more common than symptoms related to their involvement. There is a progression in the acromegalic arthropathy, resembling that described for the acromegalic cardiomyopathy or for the acromegalic features at other organs, such as thyroid, breast and prostate. At early stage, cartilage hypertrophy predominates, and then degenerative changes start until osteoarthritis features occur. Arthropathy can be reversed by normalizing GH and IGF-I levels only at the initial stage. If the disease is left untreated for long time, the osteo-articular sequelae cannot be modified by any treatment of acromegaly.

Reversal of Cirrhosis in a Patient with Primary Biliary Cirrhosis-autoimmune Hepatitis Overlap Syndrome

Digestive Diseases and Sciences. Jan, 2005  |  Pubmed ID: 15712656

Plasmapheresis Improves the Outcome of Central Pontine Myelinolysis

Journal of Neurology. Jun, 2005  |  Pubmed ID: 15742105

Allele Frequencies for 70 Autosomal SNP Loci with U.S. Caucasian, African-American, and Hispanic Samples

Forensic Science International. May, 2005  |  Pubmed ID: 15749374

189 samples from 3 different U.S. sample groups Caucasian (74), African American (71) and Hispanic (44) were typed for 70 autosomal genetic markers. These 70 markers are bi-allelic (C/T) short nucleotide polymorphisms (SNPs). For each sample, the 70 SNP markers were typed in 11 unique 6-plexes and a single 4-plex PCR. A total of 10 of the 210 tests (70 loci x 3 populations) for Hardy-Weinberg equilibrium indicated a statistically significant result. In order to evaluate the minimum number of SNP loci needed to distinguish all 189 samples from one another, we ranked the loci according to their levels of observed heterozygosity and p-values obtained upon testing for Hardy-Weinberg equilibrium. The top 12 loci according to these ranking criteria were tabulated along with the number of unique genotypes observed when combining subsequent SNP markers. The 12 selected SNPs possessed an observed heterozygosity of >0.45 in all three populations examined and thus would be expected to exhibit more differences between samples. All of the 189 samples in this study were individualized with a subset of 12 SNP loci. However, it is likely that the addition of more than 12 SNP loci will be required to resolve larger sets of unrelated individuals from one another. By way of comparison, in these same 189 individuals all but one pair is resolved from one another with three of the traditional short tandem repeat (STR) loci possessing the highest heterozygosity values (D2S1338, D18S51, and FGA) run with the Identifiler kit. The final pair of unrelated samples could be resolved with the combination of 4 STR loci: D2S1338, D18S51, FGA, and VWA.

Mitochondrial DNA Typing Screens with Control Region and Coding Region SNPs

Journal of Forensic Sciences. Mar, 2005  |  Pubmed ID: 15813549

Mitochondrial DNA (mtDNA) analysis has found an important niche in forensic DNA typing. It is used with highly degraded samples or low-copy number materials such as might be found from shed hair or bones exposed to severe environmental conditions. The primary advantage of mtDNA is that it is present in high copy number within cells and therefore more likely to be recovered from highly degraded specimens. A major disadvantage to traditional forensic mtDNA analysis is that it is time-consuming and labor-intensive to generate and review the 610 nucleotides of sequence information commonly targeted in hypervariable regions I and II (HVI and HVII) of the control region. In addition, common haplotypes exist in HVI/HVII mtDNA sequences that can reduce the ability to differentiate two unrelated samples. In this report we describe the utility of two newly available screening assays for rapid exclusion of non-matching samples. The LINEAR ARRAY mtDNA HVI/HVII Region-Sequencing Typing Kit (Roche Applied Science, Indianapolis, IN) was used to type 666 individuals from U.S. Caucasian, African American, and Hispanic groups. Processing of the LINEAR ARRAY probe panels "mito strips" was automated on a ProfiBlot workstation. Observable variation in 666 individuals is reported and frequencies of the mitotypes within and between populations are presented. Samples exhibiting the most common Caucasian mitotype were subdivided with a multiplexed amplification and detection assay using eleven single nucleotide polymorphisms in the mitochondrial genome. These types of screening assays should enable more rapid evaluation of forensic casework samples such that only samples not excluded would be subjected to further characterization through full HVI/HVII mtDNA sequence analysis.

Color-doppler Using Contrast Medium in Evaluating the Response to Neoadjuvant Treatment in Patients with Locally Advanced Breast Carcinoma

Anticancer Research. Jan-Feb, 2005  |  Pubmed ID: 15816633

The aim of our study was to assess the color-Doppler ultrasound (CDU) pattern in the analysis of neoadjuvant preoperative treatment of patients with locally advanced breast carcinoma, improvement after injection of contrast medium (Levovist) and possible correlations between morphological and vascular aspects of the neoplasm and postoperative histopathological findings.

Red Fluorescent Protein EqFP611 and Its Genetically Engineered Dimeric Variants

Journal of Biomedical Optics. Jan-Feb, 2005  |  Pubmed ID: 15847584

The red fluorescent protein (FP) eqFP611 from the sea anemone Entacmaea quadricolor shows favorable properties for applications as a molecular marker. Like other anthozoan FPs, it forms tetramers at physiological concentrations. The interactions among the monomers, however, are comparatively weak, as inferred from the dissociation into monomers in the presence of sodium dodecyl sulfate (SDS) or at high dilution. Analysis at the single-molecule level revealed that the monomers are highly fluorescent. For application as fusion markers, monomeric FPs are highly desirable. Therefore, we examine the monomer interfaces in the x-ray structure of eqFP611 to provide a basis for the rational design of monomeric variants. The arrangement of the four beta cans is very similar to that of other green fluorescent protein (GFP-like) proteins such as DsRed and RTMS5. A variety of structural features of the tetrameric interfaces explain the weak subunit interactions in eqFP611. We produce functional dimeric variants by introducing single point mutations in the A/B interface (Thr122Arg, Val124Thr). By contrast, structural manipulations in the A/C interface result in essentially complete loss of fluorescence, suggesting that A/C interfacial interactions play a crucial role in the folding of eqFP611 into its functional form.

Renoprotective Effect of Early Inhibition of the Renin-angiotensin System in Renal Transplant Recipients

Transplantation Proceedings. Mar, 2005  |  Pubmed ID: 15848600

The aim of this work was to study the effect of early administration of angiotensin-converting enzyme inhibitors (ACEI) and angiotensin II type-I receptors blockers (ARB) on renal function and proteinuria in renal transplant recipients with good, stable renal function and mild proteinuria. Twenty four patients started ACEI/ARB therapy within 14 months after surgery (RAS-). Before (T0) and every month for 2 years after the initiation of ACEI/ARB we evaluated creatinine clearance (CrCl), proteinuria/day (UP), UP/CrCl (FUP), arterial blood pressure, and serum lipid levels. Twenty-eight patients who never received ACEI/ARB (RAS+) were studied in the same fashion. In the RAS+ CrCl was reduced after 2 years compared with T0 (64.5 +/- 2.6 vs 75.0 +/- 3.2 mL/min, P < .003); UP and FUP were both significantly increased (666 +/- 65 vs 132 +/- 20 mg/day 8.8 +/- 1.2 vs 2.6 +/- 0.6 mg/mL x 10(3); P < .001 and .002) compared with T0. Moreover, UP (P < .04), FUP (P < .03), and the percentage reduction of CrCl (11.4% +/- 5% vs 4.6% +/- 1.8%; P < .05) were greater in RAS+ than RAS- subjects at 2 years of the study. The values of other parameters did not show significant differences between the two groups. In conclusion, this study suggested that ACEI/ARB have renoprotective effects, when used in patients with good stable renal function and mild proteinuria. These drugs may play a role to prevent chronic allograft nephropathy.

Molecular Dynamics Simulation of Sperm Whale Myoglobin: Effects of Mutations and Trapped CO on the Structure and Dynamics of Cavities

Biophysical Journal. Jul, 2005  |  Pubmed ID: 15849248

The results of extended (80-ns) molecular dynamics simulations of wild-type and YQR triple mutant of sperm whale deoxy myoglobin in water are reported and compared with the results of the simulation of the intermediate(s) obtained by photodissociation of CO in the wild-type protein. The opening/closure of pathways between preexistent cavities is different in the three systems. For the photodissociated state, we previously reported a clear-cut correlation between the opening probability and the presence of the photolyzed CO in the proximity of the passage; here we show that in wild-type deoxy myoglobin, opening is almost random. In wild-type deoxy myoglobin, the passage between the distal pocket and the solvent is strictly correlated to the presence/absence of a water molecule that simultaneously interacts with the distal histidine side chain and the heme iron; conversely, in the photodissociated myoglobin, the connection with the bulk solvent is always open when CO is in the vicinity of the A pyrrole ring. In YQR deoxy myoglobin, the mutated Gln(E7)64 is stably H-bonded with the mutated Tyr(B10)29. The essential dynamics analysis unveils a different behavior for the three systems. The motion amplitude is progressively restricted in going from wild-type to YQR deoxy myoglobin and to wild-type myoglobin photoproduct. In all cases, the principal motions involve mainly the same regions, but their directions are different. Analysis of the dynamics of the preexisting cavities indicates large fluctuations and frequent connections with the solvent, in agreement with the earlier hypothesis that some of the ligand may escape from the protein through these pathways.

Neuroglobin, Nitric Oxide, and Oxygen: Functional Pathways and Conformational Changes

Proceedings of the National Academy of Sciences of the United States of America. Jun, 2005  |  Pubmed ID: 15932948

Neuroglobin (Ngb) is a globin expressed in the nervous system of humans and other organisms that is involved in the protection of the brain from ischemic damage. Despite considerable interest, however, the in vivo function of Ngb is still a conundrum. In this paper we report a number of kinetic experiments with O2 and NO that we have interpreted on the basis of the 3D structure of Ngb, now available for human and murine metNgb and murine NgbCO. The reaction of reduced deoxyNgb with O2 and NO is slow (t(1/2) approximately 2 s) and ligand concentration-independent, because exogenous ligand binding can only occur upon dissociation of the distal His-64, which is coordinated to the ferrous heme iron. By contrast, NgbO2 reacts very rapidly with NO, yielding metNgb and NO3- by means of a heme-bound peroxynitrite intermediate. Steady-state amperometric experiments show that Ngb is devoid of O2 reductase and NO reductase activities. To achieve this result, we have set up a protocol for efficient reduction of metNgb using a mixture of FMN and NADH under bright illumination. The results are discussed with reference to a global scheme inspired by the 3D structures of metNgb and NgbCO. Based on the ligand-linked conformational changes discovered by crystallography, the pathways of the reactions with O2 and NO provide a framework that may account for the involvement of Ngb in controlling the activation of a protective signaling mechanism.

Significant Long-term Survival After Radiofrequency Ablation of Unresectable Hepatocellular Carcinoma in Patients with Cirrhosis

Annals of Surgical Oncology. Aug, 2005  |  Pubmed ID: 15965731

Radiofrequency ablation (RFA) offers an alternative treatment in some unresectable hepatocellular carcinoma (HCC) patients with disease confined to the liver. We prospectively evaluated survival rates in patients with early-stage, unresectable HCC treated with RFA.

Cystic Lymphangioma Associated with Enteric Duplication As a Cause of Recurrent Vomiting

Digestive and Liver Disease : Official Journal of the Italian Society of Gastroenterology and the Italian Association for the Study of the Liver. Jul, 2005  |  Pubmed ID: 15975542

We describe a case report of a 6-year-old boy with a 4-year history of recurrent vomiting with a cyclical vomiting pattern. Although initially labelled with and treated for Cyclical Vomiting Syndrome the cause was subsequently found to be an enteric duplication associated with cystic lymphangioma, an association not previously described.

Biweekly Docetaxel-irinotecan Treatment with Filgrastim Support is Highly Active in Antracycline-Paclitaxel-refractory Breast Cancer Patients

Oncology. 2005  |  Pubmed ID: 16020968

To evaluate the feasibility and activity of combination treatment with docetaxel (DTX) and irinotecan (CPT-11), given together every other week, combined with filgrastim support, in anthracycline- and paclitaxel-pretreated breast cancer (BC) patients.

Chromosomal Duplications Along the Y-chromosome and Their Potential Impact on Y-STR Interpretation

Journal of Forensic Sciences. Jul, 2005  |  Pubmed ID: 16078487

Y-chromosome short tandem repeat (Y-STR) markers are being used as potential tools for distinguishing low levels of male DNA in the presence of excess female DNA as is present in many sexual assault samples. Usually single copy Y-STR loci produce a single amplicon in single source samples, and thus the observation of multiple peaks at such a locus could suggest to an analyst that a mixture of more than one male contributor is present in the tested sample. However, many regions of the Y-chromosome are duplicated or even triplicated in some individuals and this fact can thus complicate potential mixture interpretation. Reasons for the presence of duplications at multiple loci within a single sample are explored in the context of Y-STR marker location along the chromosome. True male-male mixtures commonly exhibit more than one locus-specific PCR product across multiple Y-STR loci that are not adjacent to one another on the Y-chromosome. In addition, duplicated loci typically possess alleles that differ by only a single repeat unit and possess similar peak heights.

Ultrasound Contrast Media in the Study of Salivary Gland Tumors

Anticancer Research. May-Jun, 2005  |  Pubmed ID: 16080479

The aims of the present study were: to assess standard ultrasound and color-Doppler patterns for the study and characterization of salivary gland tumors; to define the improvement in the color-Doppler ultrasound image after injection of a signal amplifier (Levovist); to compare morphological and vascular aspects of the neoplasm with data obtained during postoperative histopathological analysis.

Long-term Follow-up of Lupus-related Subacute Myelopathy Treated by Plasmapheresis and Pulse Cyclophosphamide

Clinical and Experimental Rheumatology. Sep-Oct, 2005  |  Pubmed ID: 16173268

Temperature Regulates Transcription in the Zebrafish Circadian Clock

PLoS Biology. Nov, 2005  |  Pubmed ID: 16176122

It has been well-documented that temperature influences key aspects of the circadian clock. Temperature cycles entrain the clock, while the period length of the circadian cycle is adjusted so that it remains relatively constant over a wide range of temperatures (temperature compensation). In vertebrates, the molecular basis of these properties is poorly understood. Here, using the zebrafish as an ectothermic model, we demonstrate first that in the absence of light, exposure of embryos and primary cell lines to temperature cycles entrains circadian rhythms of clock gene expression. Temperature steps drive changes in the basal expression of certain clock genes in a gene-specific manner, a mechanism potentially contributing to entrainment. In the case of the per4 gene, while E-box promoter elements mediate circadian clock regulation, they do not direct the temperature-driven changes in transcription. Second, by studying E-box-regulated transcription as a reporter of the core clock mechanism, we reveal that the zebrafish clock is temperature-compensated. In addition, temperature strongly influences the amplitude of circadian transcriptional rhythms during and following entrainment by light-dark cycles, a property that could confer temperature compensation. Finally, we show temperature-dependent changes in the expression levels, phosphorylation, and function of the clock protein, CLK. This suggests a mechanism that could account for changes in the amplitude of the E-box-directed rhythm. Together, our results imply that several key transcriptional regulatory elements at the core of the zebrafish clock respond to temperature.

Effects of Successful Renal Transplantation on Left Ventricular Mass

Transplantation Proceedings. Jul-Aug, 2005  |  Pubmed ID: 16182718

Left ventricular hypertrophy is an independent cardiovascular risk factor in the general population and in patients with chronic renal failure. Relatively little is known about the effects of renal transplantation on left ventricular hypertrophy. The aim of this study was to determine the changes in left ventricular mass after successful renal transplantation and to evaluate the importance of some clinical, laboratory, and echocardiographic variables on the trend to left ventricular hypertrophy. Twenty-three patients with end-stage renal disease were studied by ambulatory blood pressure monitoring and echocardiography before and 2 years following renal transplantation. After 24 months of follow-up, all transplant recipients had adequate renal function (serum creatinine <2 mg/dL). At the end of the study, we observed a significant decrease in left ventricular mass and left ventricular mass index compared to the pretransplantation period. In renal transplant recipients, the prevalence of left ventricular hypertrophy significantly decreased (78% versus 44%, P < .03) after 2 years of follow-up. Systolic 24-hour blood pressure was the only predictor of left ventricular mass and of left ventricular mass index at 2 years after transplantation. In conclusion, successful renal transplantation produces a regression of left ventricular hypertrophy. This beneficial effect depends on a decrease in systolic pressure levels.

Effect of the New York State Cigarette Fire Safety Standard on Ignition Propensity, Smoke Constituents, and the Consumer Market

Tobacco Control. Oct, 2005  |  Pubmed ID: 16183983

This study examines empirical evidence from the New York experience testing tobacco industry arguments made in opposition to fire safety standards for cigarettes.

Zebrafish Cell Clocks Feel the Heat and See the Light!

Zebrafish. 2005  |  Pubmed ID: 18248192

The zebrafish has rapidly become established as one of the most valuable vertebrate models for studying circadian clock function. A major initial attraction was its utility in large-scale genetic screens. It subsequently emerged that most zebrafish cells possess circadian clocks that can be entrained directly by exposure to temperature or light dark cycles, a property shared by several zebrafish cell lines. This is not the case for mammals, where the retina is the primary source of light input to the clock. Furthermore, mammalian cell culture clocks can only be entrained by acute culture treatments such as serum shocks. Thus, the zebrafish is proving invaluable to study light and temperature input to the vertebrate clock. In addition, the accessibility of its early developmental stages has placed the zebrafish at the forefront of studies aimed at understanding how the circadian clock is established during embryogenesis.

Pulsating Enophthalmos in an Adult Patient with Type 1 Neurofibromatosis

Neurology. Dec, 2006  |  Pubmed ID: 17190938

[Current Trend in the Diagnosis and Treatment of Primary Non Refluxing Megaureter]

La Pediatria Medica E Chirurgica : Medical and Surgical Pediatrics. 2006  |  Pubmed ID: 17533904

The terms megaureter or hydroureteronephrosis are non-specific because indicate various pathologic entities recognise different causes (obstruction, reflux, obstruction-reflux, primary and secondary). An undeveloped renal function in neonatal period makes more difficult the therapeutic approach. Actually the problem is to find the indicators that consent us the individualization of patients more suitable for nonoperative management.

Special Effects: Tobacco Policies and Low Socioeconomic Status Girls and Women

Journal of Epidemiology and Community Health. Sep, 2006  |  Pubmed ID: 17708002

Effective Strategies for Forensic Analysis in the Mitochondrial DNA Coding Region

International Journal of Legal Medicine. Jan, 2006  |  Pubmed ID: 16261373

Recently, it has been recognized that accessing information in the mtDNA coding region can provide additional forensic discrimination with respect to the standard typing of the D-loop region, augmenting the sometimes rather limited forensic power of mtDNA testing. Here, we discuss considerations relating to maximally effective approaches for recovering additional discrimination in the coding region, bearing in mind that (1) DNA quality and quantity in typical mtDNA casework usually restrict the amount of additional sequence that can be obtained, and (2) the need for additional discrimination primarily arises when common HV1/HV2 types are encountered. Most investigators have sought additional discrimination by sequencing short segments of coding region that are thought to be particularly variable. Unfortunately, efforts in this regard have generally failed to appreciate that most variation in the coding region is redundant with information already present in HV1/HV2 and have therefore overvalued the potential of this approach for providing additional discrimination. An alternative single nucleotide polymorphism-based approach [Int J Legal Med 118:137-146, 2004] has been to identify specific bases that provide resolution in specific common HV1/HV2 types (and related sequences). We investigate several highly relevant data sets wherein the latter approach performs appreciably better than sequencing selected short portions of the coding region. This is true even when only synonymous variation is targeted to minimize the potential for problems arising from discovery of mutations that have reportedly been related to disease.

Analysis of Artificially Degraded DNA Using STRs and SNPs--results of a Collaborative European (EDNAP) Exercise

Forensic Science International. Dec, 2006  |  Pubmed ID: 16343834

Recently, there has been much debate about what kinds of genetic markers should be implemented as new core loci that constitute national DNA databases. The choices lie between conventional STRs, ranging in size from 100 to 450 bp; mini-STRs, with amplicon sizes less than 200 bp; and single nucleotide polymorphisms (SNPs). There is general agreement by the European DNA Profiling Group (EDNAP) and the European Network of Forensic Science Institutes (ENFSI) that the reason to implement new markers is to increase the chance of amplifying highly degraded DNA rather than to increase the discriminating power of the current techniques. A collaborative study between nine European and US laboratories was organised under the auspices of EDNAP. Each laboratory was supplied with a SNP multiplex kit (Foren-SNPs) provided by the Forensic Science Service, two mini-STR kits provided by the National Institute of Standards and Technology (NIST) and a set of degraded DNA stains (blood and saliva). Laboratories tested all three multiplex kits, along with their own existing DNA profiling technique, on the same sets of degraded samples. Results were collated and analysed and, in general, mini-STR systems were shown to be the most effective. Accordingly, the EDNAP and ENFSI working groups have recommended that existing STR loci are reengineered to provide smaller amplicons, and the adoption of three new European core loci has been agreed.

Studies of DNA Dumbbells VIII. Melting Analysis of DNA Dumbbells with Dinucleotide Repeat Stem Sequences

Biopolymers. Jun, 2006  |  Pubmed ID: 16345003

Melting curves and circular dichroism spectra were measured for a number of DNA dumbbell and linear molecules containing dinucleotide repeat sequences of different lengths. To study effects of different sequences on the melting and spectroscopic properties, six DNA dumbbells whose stems contain the central sequences (AA)(10), (AC)(10), (AG)(10), (AT)(10), (GC)(10), and (GG)(10) were prepared. These represent the minimal set of 10 possible dinucleotide repeats. To study effects of dinucleotide repeat length, dumbbells with the central sequences (AG)(n), n = 5 and 20, were prepared. Control molecules, dumbbells with a random central sequence, (RN)(n), n = 5, 10, and 20, were also prepared. The central sequence of each dumbbell was flanked on both sides by the same 12 base pairs and T(4) end-loops. Melting curves were measured by optical absorbance and differential scanning calorimetry in solvents containing 25, 55, 85, and 115 mM Na(+). CD spectra were collected from 20 to 45 degrees C and [Na(+)] from 25 to 115 mM. The spectral database did not reveal any apparent temperature dependence in the pretransition region. Analysis of the melting thermodynamics evaluated as a function of Na(+) provided a means for quantitatively estimating the counterion release with melting for the different sequences. Results show a very definite sequence dependence, indicating the salt-dependent properties of duplex DNA are also sequence dependent. Linear DNA molecules containing the (AG)(n) and (RN)(n), sequences, n = 5, 10, 20, and 30, were also prepared and studied. The linear DNA molecules had the exact sequences of the dumbbell stems. That is, the central repeat sequence in each linear duplex was flanked on both sides by the same 12-bp sequence. Melting and CD studies were also performed on the linear DNA molecules. Comparison of results obtained for the same sequences in dumbbell and linear molecular environments reveals several interesting features of the interplay between sequence-dependent structural variability, sequence length, and the unconstrained (linear) or constrained (dumbbell) molecular environments.

Combined Use of Noninvasive Tests is Useful in the Initial Diagnostic Approach to a Child with Suspected Inflammatory Bowel Disease

Journal of Pediatric Gastroenterology and Nutrition. Jan, 2006  |  Pubmed ID: 16385247

To assess the effectiveness of the combined use of fecal calprotectin (FC), anti-Saccharomyces cerevisiae antibody (ASCA), perinuclear staining antineutrophil antibody (pANCA), small intestinal permeability test (IP), and bowel wall ultrasonography measurement (BWUS) in the diagnostic work-up of children with suspected inflammatory bowel disease (IBD).

Allele Frequencies for 27 Y-STR Loci with U.S. Caucasian, African American, and Hispanic Samples

Forensic Science International. Jan, 2006  |  Pubmed ID: 16410169

A total of 263 U.S. Caucasians, 260 African Americans and 140 U.S. Hispanics or a subset of 31 Caucasians, 32 African Americans, and 32 Hispanics were typed for 27 Y-chromosome short tandem repeat (Y-STR) markers: DYS444, DYS446, DYS449, DYS463, DYS485, DYS490, DYS495, DYS504, DYS505, DYS508, DYS520, DYS522, DYS525, DYS532, DYS533, DYS534, DYS540, DYS556, DYS557, DYS570, DYS575, DYS576, DYS594, DYS632, DYS635, DYS641, and DYS643. Allele frequencies for each locus are reported along with nomenclature based on sequence analysis.

The Allosteric Properties of Hemoglobin: Insights from Natural and Site Directed Mutants

Current Protein & Peptide Science. Feb, 2006  |  Pubmed ID: 16472167

After over a century of extensive research, hemoglobin has become the prototype of allosteric and cooperative proteins. Its molecular structure, known in great detail, has allowed the design of hundreds of site directed mutations, aimed at interfering with its function, and thus at testing our hypotheses on the molecular mechanisms of allostery. The wealth of information thus obtained is difficult to read except for specialists, not only because it makes use of many different technical approaches, but also because of its intrinsically patchy nature. Moreover, several researchers have tried to assign specific roles to segments of the polypeptide chains, rather than to single residues, and have tested their hypotheses by multiple point mutations or by complete replacement with the homologous segment from a different hemoglobin to produce chimeric macromolecules. This approach is in great need of a revision since putative functionally relevant segments partially overlap. This review briefly describes the structure and function of hemoglobin, and analyzes the effect of point mutations, multiple mutations and segment replacement, with special attention to possible biotechnological applications, ranging from pharmacology (Hb solutions as resuscitating fluids and sources of the protein found in hemoglobinopathies for biochemical studies) to bioreactors. Occasional reference is made to site directed mutants of myoglobin, whenever this helps clarifying perplexing results obtained on hemoglobin.

Perspectives from the Front Lines of Tobacco Control

Journal of Health Care for the Poor and Underserved. Feb, 2006  |  Pubmed ID: 16520521

This research is designed to share valuable experiences and transferable principles from program staff of the Legacy/Community Voices initiative who have been involved in planning, implementing, evaluating, and sustaining tobacco control activities in underserved communities. Interviews were conducted with 13 front line staff from 9 sites: Alameda County, California; Detroit, Michigan; El Paso, Texas; Ingham County, Michigan; Miami, Florida; New Mexico; North Carolina; Northern Manhattan; and West Virginia. A model emerged from these interviews that places the life cycle of a program in a central position, with many of the identified themes (working with local champions, obtaining support from multiple partners, increasing organizational capacity) repeated throughout, albeit in different forms at different stages. Reflecting upon wisdom gained and identifying best processes for such work may help ensure that tobacco control programs are developed that are culturally safe and effective in meeting the needs of diverse communities throughout the United States.

Combined Ethanol Injection Therapy and Radiofrequency Ablation Therapy in Percutaneous Treatment of Hepatocellular Carcinoma Larger Than 4 Cm

Cardiovascular and Interventional Radiology. Jul-Aug, 2006  |  Pubmed ID: 16528626

Optimal treatment of large-sized hepatocellular carcinoma (HCC) is still debated, because percutaneous ablation therapies alone do not always achieve complete necrosis.

Extended Subnanosecond Structural Dynamics of Myoglobin Revealed by Laue Crystallography

Proceedings of the National Academy of Sciences of the United States of America. Mar, 2006  |  Pubmed ID: 16547137

Work carried out over the last 30 years unveiled the role of structural dynamics in controlling protein function. Cavity networks modulate structural dynamics trajectories and are functionally relevant; in globins they have been assigned a role in ligand migration and docking. These findings raised renewed interest for time-resolved structural investigations of myoglobin (Mb), a simple heme protein displaying a photosensitive iron-ligand bond. Photodissociation of MbCO generates a nonequilibrium population of protein structures relaxing over a time range extending from picoseconds to milliseconds. This process triggers ligand migration to matrix cavities with clear-cut effects on the rate and yield of geminate rebinding. Here, we report subnanosecond time-resolved Laue diffraction data on the triple mutant YQR-Mb [Leu-29(B10)Tyr, His-64(E7)Gln, Thr-67(E10)Arg] that depict the sequence of structural events associated with heme and protein relaxation from 100 ps to 316 ns and above. The photodissociated ligand rapidly (<0.1 ns) populates the Xe-binding cavity distal to the heme. Moreover, the heme relaxation toward the deoxy configuration is heterogeneous, with a slower phase ( approximately ns) evident in these experiments. Damping of the heme response appears to result from a strain exerted by the E-helix via the CD-turn; Phe-43(CD1), in close contact with heme, opposes tilt until the strain is relieved. A comparison with crystallographic data on wild-type Mb and mutants Leu(29)Phe or Leu(29)Trp suggests that the internal structure controls the rate and amplitude of the relaxation events. A correlation between structural dynamics as unveiled by Laue crystallography and functional properties of Mb is presented.

Zebrafish Arylalkylamine-N-acetyltransferase Genes - Targets for Regulation of the Circadian Clock

Journal of Molecular Endocrinology. Apr, 2006  |  Pubmed ID: 16595704

Daily rhythms of melatonin production are controlled by changes in the activity of arylalkylamine-N-acetyltransferase (AANAT). Zebrafish possess two aanats, aanat1 and aanat2; the former is expressed only in the retina and the latter is expressed in both the retina and the pineal gland. Here, their differential expression and regulation were studied using transcript quantification and transient and stable in vivo and in vitro transfection assays. In the pineal gland, the aanat2 promoter exhibited circadian clock-controlled activity, as indicated by circadian rhythms of Enhanced green fluorescent protein (EGFP) mRNA in AANAT2:EGFP transgenic fish. In vivo transient expression analyses of the aanat2 promoter indicated that E-box and photoreceptor conserved elements (PCE) are required for expression in the pineal gland. In the retina, the expression of both genes was characterized by a robust circadian rhythm of their transcript levels. In constant darkness, the rhythmic expression of retinal aanat2 persisted while the aanat1 rhythm disappeared; indicating that the former is controlled by a circadian clock and the latter is also light driven. In the light-entrainable clock-containing PAC-2 zebrafish cell line, both stably transfected aanat1 and aanat2 promoters exhibited a clock-controlled circadian rhythm, characteristic for an E-box-driven expression. Transient co-transfection experiments in NIH-3T3 cells revealed that the two, E-box- and PCE-containing, promoters are driven by the synergistic action of BMAL/CLOCK and orthehodenticle homeobox 5. This study has revealed a shared mechanism for the regulation of two related genes, yet describes their differential phases and photic responses which may be driven by other gene-specific regulatory mechanisms and tissue-specific transcription factor profiles.

Photoconvertible Fluorescent Protein EosFP: Biophysical Properties and Cell Biology Applications

Photochemistry and Photobiology. Mar-Apr, 2006  |  Pubmed ID: 16613485

EosFP is a fluorescent protein from the coral Lobophyllia hemprichii that changes its fluorescence emission from green to red upon irradiation with near-UV light. Here we present the spectroscopic properties of wild-type EosFP and a variety of monomeric and dimeric mutants and provide a structural interpretation of its oligomerization and photoconversion, which is based on X-ray structure analysis of the green and red species that we reported recently. Because functional expression of the monomeric EosFP variant is limited to temperatures of 30 degrees C, we have developed a tandem dimer. This construct, in which two EosFP subunits are connected by a flexible 12 amino acid linker, expresses well after fusion with the androgen and endothelin A receptors at 37 degrees C. A variety of applications in cellular imaging, developmental biology and automated high-content screening applications are presented, which demonstrate that EosFP is a powerful tool for in vivo monitoring of cellular processes.

Molecular Analysis of Clock Gene Expression in the Avian Brain

Chronobiology International. 2006  |  Pubmed ID: 16687285

Birds are equipped with a complex circadian pacemaking system that regulates the rhythmicity of physiology and behavior. As with all organisms, transcriptional and translational feedback loops of clock genes represent the basic molecular mechanism of rhythm generation in birds. To investigate avian clock gene expression, partial cDNA sequences of six mammalian clock gene homologs (Bmal1, Clock, Per2, Per3, Cry1, and Cry2) and a novel avian cryptochrome gene (Cry4) were cloned from the house sparrow, a model system in circadian research. Expression patterns were analyzed by semi-quantitative RT-PCR and RNase protection assays using total RNA extracted from adult male house sparrow brains. With the exception of Cry4, pronounced rhythmic mRNA expression of all the clock genes analyzed was encountered, with mRNA levels varying considerably between the various genes. Although some basic features of the molecular circadian feedback loop appear to be similar between mammals and birds, the precise phase relationships of the clock gene mRNA rhythms relative to each other and to the light zeitgeber differ significantly between the house sparrow and mammals. Our results point to the existence of differences in the organization of avian and mammalian circadian clock mechanisms.

Isolation and Characterization of Melanopsin and Pinopsin Expression Within Photoreceptive Sites of Reptiles

Die Naturwissenschaften. Aug, 2006  |  Pubmed ID: 16688437

Non-mammalian vertebrates have multiple extraocular photoreceptors, mainly localised in the pineal complex and the brain, to mediate irradiance detection. In this study, we report the full-length cDNA cloning of ruin lizard melanopsin and pinopsin. The high level of identity with opsins in both the transmembrane regions, where the chromophore binding site is located, and the intracellular loops, where the G-proteins interact, suggests that both melanopsin and pinopsin should be able to generate a stable photopigment, capable of triggering a transduction cascade mediated by G-proteins. Phylogenetic analysis showed that both opsins are located on the expected branches of the corresponding sequences of ortholog proteins. Subsequently, using RT-PCR and RPA analysis, we verified the expression of ruin lizard melanopsin and pinopsin in directly photosensitive organs, such as the lateral eye, brain, pineal gland and parietal eye. Melanopsin expression was detected in the lateral eye and all major regions of the brain. However, different from the situation in Xenopus and chicken, melanopsin is not expressed in the ruin lizard pineal. Pinopsin mRNA expression was only detected in the pineal complex. As a result of their phylogenetic position and ecology, reptiles provide the circadian field with some of the most interesting models for understanding the evolution of the vertebrate circadian timing system and its response to light. This characterization of melanopsin and pinopsin expression in the ruin lizard will be important for future studies aimed at understanding the molecular basis of circadian light detection in reptiles.

Smoking, Obesity, and Their Co-occurrence in the United States: Cross Sectional Analysis

BMJ (Clinical Research Ed.). Jul, 2006  |  Pubmed ID: 16698804

To describe the prevalence of obesity, smoking, and both health risk factors together among adults in the United States.

A Globin for the Brain

FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology. Nov, 2006  |  Pubmed ID: 17077295

The discovery that a myoglobin-like hemeprotein (called neuroglobin) is expressed in our brain raised considerable curiosity from the standpoints of biochemistry and pathophysiology alike. Neuroglobin is involved in neuroprotection from damage due to hypoxia or ischemia in vitro and in vivo; overexpression of neuroglobin ameliorates the recovery from stroke in experimental animals. The mechanism underlying this remarkable effect is still mysterious. Structural studies revealed that neuroglobin has a typical globin fold, and despite being hexacoordinated, it binds reversibly O2, CO, and NO, undergoing a substantial conformational change of the heme and of the protein. The possible mechanisms involved in neuroprotection are briefly reviewed. Neuroglobin is unlikely to be involved in O2 transport (like myoglobin), although it seems to act as a sensor of the O2/NO ratio in the cell, possibly regulating the GDP/GTP exchange rate forming a specific complex with the G(alpha beta gamma)-protein when oxidized but not when bound to a gaseous ligand. Thus it appears that neuroglobin is a stress-responsive sensor for signal transduction in the brain, mediated by a ligand-linked conformational change of the protein.

The Structure of the Endoribonuclease XendoU: From Small Nucleolar RNA Processing to Severe Acute Respiratory Syndrome Coronavirus Replication

Proceedings of the National Academy of Sciences of the United States of America. Aug, 2006  |  Pubmed ID: 16895992

Small nucleolar RNAs (snoRNAs) play a key role in eukaryotic ribosome biogenesis. In most cases, snoRNAs are encoded in introns and are released through the splicing reaction. Some snoRNAs are, instead, produced by an alternative pathway consisting of endonucleolytic processing of pre-mRNA. XendoU, the endoribonuclease responsible for this activity, is a U-specific, metal-dependent enzyme that releases products with 2'-3' cyclic phosphate termini. XendoU is broadly conserved among eukaryotes, and it is a genetic marker of nidoviruses, including the severe acute respiratory syndrome coronavirus, where it is essential for replication and transcription. We have determined by crystallography the structure of XendoU that, by refined search methodologies, appears to display a unique fold. Based on sequence conservation, mutagenesis, and docking simulations, we have identified the active site. The conserved structural determinants of this site may provide a framework for attempting to design antiviral drugs to interfere with the infectious nidovirus life cycle.

Endocrine Dysfunction in Patients with Fabry Disease

The Journal of Clinical Endocrinology and Metabolism. Nov, 2006  |  Pubmed ID: 16926253

Fabry disease (FD) is a genetic disorder caused by lysosomal alpha-galactosidase-A deficiency and is characterized by the systemic accumulation of globotriaosylceramide. All endocrine glands are susceptible to globotriaosylceramide accumulation because of their high vascularization and low cellular proliferation rate. Nevertheless, this endocrine system has never been investigated in detail.

Televised Movie Trailers: Undermining Restrictions on Advertising Tobacco to Youth

Archives of Pediatrics & Adolescent Medicine. Sep, 2006  |  Pubmed ID: 16953010

To determine the proportion of televised movie trailers that included images of tobacco use during 1 year and the extent of youth exposure to those trailers.

Chromophore-protein Interactions in the Anthozoan Green Fluorescent Protein AsFP499

Biophysical Journal. Dec, 2006  |  Pubmed ID: 16980366

Despite their similar fold topologies, anthozoan fluorescent proteins (FPs) can exhibit widely different optical properties, arising either from chemical modification of the chromophore itself or from specific interactions of the chromophore with the surrounding protein moiety. Here we present a structural and spectroscopic investigation of the green FP asFP499 from the sea anemone Anemonia sulcata var. rufescens to explore the effects of the protein environment on the chromophore. The optical absorption and fluorescence spectra reveal two discrete species populated in significant proportions over a wide pH range. Moreover, multiple protonation reactions are evident from the observed pH-dependent spectral changes. The x-ray structure of asFP499, determined by molecular replacement at a resolution of 1.85 A, shows the typical beta-barrel fold of the green FP from Aequorea victoria (avGFP). In its center, the chromophore, formed from the tripeptide Gln(63)-Tyr(64)-Gly(65), is tightly held by multiple hydrogen bonds in a polar cage that is structurally quite dissimilar to that of avGFP. The x-ray structure provides interesting clues as to how the spectroscopic properties are fine tuned by the chromophore environment.

Contrast-enhanced Sonography of the Spleen

Seminars in Ultrasound, CT, and MR. Oct, 2006  |  Pubmed ID: 17048457

Sonography is frequently the first imaging modality employed to evaluate the spleen. Nevertheless, splenic abnormalities frequently determine subtle changes in echogenicity and may be overlooked. Additionally, splenic disorders have a nonspecific appearance, mostly appearing as hypoechoic defects. Difficulties in differential diagnosis are not uncommon, with splenic infarctions resembling, for example, focal lesions. Color Doppler is usually of limited additional value. Instead, the spleen is optimally suited for contrast-enhanced sonography, being superficial, small, and homogeneous, and showing intense and persistent contrast enhancement. Scarcely evident abnormalities become evident after contrast medium injection. Additional information can be obtained in many clinical scenarios, including blunt trauma, left flank pain, lymphoma, and incidental detection of splenic abnormalities.

Exploring Chromophore--protein Interactions in Fluorescent Protein CmFP512 from Cerianthus Membranaceus: X-ray Structure Analysis and Optical Spectroscopy

Biochemistry. Oct, 2006  |  Pubmed ID: 17059211

Autofluorescent proteins of the GFP family all share the same three-dimensional beta-can fold; yet they exhibit widely different optical properties, arising either from chemical modification of the chromophore itself or from specific interactions of the chromophore with the surrounding protein moiety. Here we present a structural and spectroscopic characterization of the green fluorescent protein cmFP512 from Cerianthus membranaceus, a nonbioluminescent, azooxanthellate cnidarian, which has only approximately 22% sequence identity with Aequorea victoria GFP. The X-ray structure, obtained by molecular replacement at a resolution of 1. 35 A, shows the chromophore, formed from the tripeptide Gln-Tyr-Gly, in a hydrogen-bonded cage in the center of an 11-stranded beta-barrel, tightly restrained by adjacent residues and structural water molecules. It exists in a neutral (A) and an anionic (B) species, with absorption/emission maxima at 392/460 (pH 5) and 503/512 nm (pH 7). Their fractional populations and peak positions depend sensitively on pH, reflecting protonation of groups adjacent to the chromophore. The pH dependence of the spectra is explained by a protonation mechanism involving a hydrogen-bonded cluster of charged/polar groups. Cryospectroscopy at 12 K was also performed to analyze the vibronic coupling of the electronic transitions.

Findings and Implications from a National Study on Potential Reduced Exposure Products (PREPs)

Nicotine & Tobacco Research : Official Journal of the Society for Research on Nicotine and Tobacco. Dec, 2006  |  Pubmed ID: 17132527

Tobacco companies have recently introduced products that they claim have reduced toxins and carcinogens, and that they say may be less harmful to smokers. These are potential reduced exposure products, or PREPs. This study measured smokers' awareness of PREPs, use of PREPs, interest in trying PREPs, and beliefs about the regulation of PREPs. This study was based on nationally representative data collected in 2002 and 2003 through the American Smoking and Health Survey. The final sample included 1,174 adult smokers. Descriptive and multivariate analyses were conducted to produce estimates and explore potential correlates of the outcomes. A total of 41.9% of adult smokers reported having heard of at least one of the PREPs measured, and 11.0% reported having tried one of these products. Half of adult smokers (49.9%) said they would like to try PREPs. Interest in trying PREPs was associated with having made a quit attempt, being concerned about the effect of smoking on one's health, and having a household income of less than US dollars 20,000. About half of adult smokers (49.1%) incorrectly believed that PREPs are evaluated for safety by the government before being placed on the market, and 84.2% believed that the government should evaluate the safety of PREPs before they are sold to consumers. This study provides new and timely information on the use of, interest in trying, and beliefs about the regulation of PREPs among a nationally representative sample of adult smokers. With half of adult smokers interested in trying PREPs, the need for concrete scientific evidence on the potential impact of these products is critical.

Enhancing the Violation of the Einstein-podolsky-rosen Local Realism by Quantum Hyperentanglement

Physical Review Letters. Oct, 2006  |  Pubmed ID: 17155228

Mermin's observation [Phys. Rev. Lett. 65, 1838 (1990)] that the magnitude of the violation of local realism, defined as the ratio between the quantum prediction and the classical bound, can grow exponentially with the size of the system is demonstrated using two-photon hyperentangled states entangled in polarization and path degrees of freedom, and local measurements of polarization and path simultaneously.

Genetic Relationships Between Cercospora Kikuchii Populations from South America and Japan

Phytopathology. Sep, 2006  |  Pubmed ID: 18944056

ABSTRACT A collection 160 isolates of Cercospora kikuchii was made from South America and 245 from Japan. DNA fingerprint patterns were analyzed based on amplified fragment length polymorphism among the sample isolates, dividing the isolates into seven lineages (I to VII). Partial nucleotide sequence analyses of the beta-tubulin gene supported this division into seven lineages. Lineages I and III commonly existed in South America and Japan. In all, 136 of the 160 isolates from South America and 223 of the 245 isolates from Japan belonged to lineage I, indicating that lineage I was the major lineage in each area; 5 isolates from South America and 8 isolates from Japan belonged to lineage III. Lineages II (12 isolates) and IV (2 isolates) were specific to Japan and lineages V (3 isolates), VI (1 isolate), and VII (15 isolates) specifically existed in South America. These results suggest that the population genetic structure of C. kikuchii was different between South America and Japan, but the dominance of lineage I was common between the two areas.

Transcriptional Regulation of Arylalkylamine-N-acetyltransferase-2 Gene in the Pineal Gland of the Gilthead Seabream

Journal of Neuroendocrinology. Jan, 2007  |  Pubmed ID: 17184485

Pineal serotonin-N-acetyltransferase (arylalkylamine-N-acetyltransferase; AANAT) is considered the key enzyme in the generation of circulating melatonin rhythms; the rate of melatonin production is determined by AANAT activity. In all the examined species, AANAT activity is regulated at the post-translational level and, to a variable degree, also at the transcriptional level. Here, the transcriptional regulation of pineal aanat (aanat2) of the gilthead seabream (Sparus aurata) was investigated. Real-time polymerase chain reaction quantification of aanat2 mRNA levels in the pineal gland collected throughout the 24-h cycle revealed a rhythmic expression pattern. In cultured pineal glands, the amplitude was reduced, but the daily rhythmic expression pattern was maintained under constant illumination, indicating a circadian clock-controlled regulation of seabream aanat2. DNA constructs were prepared in which green fluorescent protein was driven by the aanat2 promoters of seabream and Northern pike. In vivo transient expression analyses in zebrafish embryos indicated that these promoters contain the necessary elements to drive enhanced expression in the pineal gland. In the light-entrainable clock-containing PAC-2 zebrafish cell line, a stably transfected seabream aanat2 promoter-luciferase DNA construct exhibited a clock-controlled circadian rhythm of luciferase activity, characteristic for an E-box-driven expression. In NIH-3T3 cells, the seabream aanat2 promoter was activated by a synergistic action of BMAL/CLOCK and orthodenticle homeobox 5 (OTX5). Promoter sequence analyses revealed the presence of the photoreceptor conserved element and an extended E-box (i.e. the binding sites for BMAL/CLOCK and OTX5 that have been previously associated with pineal-specific and rhythmic gene expression). These results suggest that seabream aanat2 is a clock-controlled gene that is regulated by conserved mechanisms.

Anatomic Excision of Anterior Prostatic Fat at Radical Prostatectomy: Implications for Pathologic Upstaging

Urology. Nov, 2007  |  Pubmed ID: 18068462

After exposure of the retropubic space, the surgeon commonly dissects the fat overlying the prostate and usually discards it. We have previously described the importance of dissecting this fat to completely visualize the dorsal venous complex (DVC) and prostatic apex. In this study, we describe a technique to dissect and remove the anterior prostatic fat pad (APF) and its anatomic and pathologic significance.

Forensic Application of the Affymetrix Human Mitochondrial Resequencing Array

Forensic Science International. Genetics. Jun, 2007  |  Pubmed ID: 19083755

In the field of forensic DNA testing, sequencing regions of the mitochondrial genome is performed when insufficient genomic DNA is present for traditional autosomal short tandem repeat (STR) testing. Sequencing coding region polymorphisms in the mitochondrial genome can be useful for resolving individuals who have the identical HV1 and HV2 control region sequence. Various methods and strategies have been established to interrogate coding region polymorphisms. These range from SNP assays probing sites most likely to differentiate individuals based on their HV1/HV2 sequence to the use of mass spectrometry to pyrosequencing. Here we evaluate the potential of the Affymetrix GeneChip Mitochondrial Resequencing Array (version 2.0) for forensic applications.

The Impact of Additional Y-STR Loci on Resolving Common Haplotypes and Closely Related Individuals

Forensic Science International. Genetics. Jun, 2007  |  Pubmed ID: 19083761

Commercial Y-STR kits have permitted laboratories to go beyond the original nine minimal haplotype loci (MHL) and to discover the advantage of additional Y-STR loci in resolving common haplotypes. In an effort to examine the impact of Y-STR markers beyond the 17 loci now available in commercial kit form, new Y-STR loci are being investigated on a common set of samples representative of the major U.S. population groups. Additional Y-STRs can also increase the power of discrimination between closely related male individuals, which is important not only in forensics but also in the paternity and genetic genealogy communities.

Start the Clock! Circadian Rhythms and Development

Developmental Dynamics : an Official Publication of the American Association of Anatomists. Jan, 2007  |  Pubmed ID: 17075872

The contribution of timing cues from the environment to the coordination of early developmental processes is poorly understood. The day-night cycle represents one of the most important, regular environmental changes that animals are exposed to. A key adaptation that allows animals to anticipate daily environmental changes is the circadian clock. In this review, we aim to address when a light-regulated circadian clock first emerges during development and what its functions are at this early stage. In particular, do circadian clocks regulate early developmental processes? We will focus on results obtained with Drosophila and vertebrates, where both circadian clock and developmental control mechanisms have been intensively studied.

Nrf2 Gene Deletion Fails to Alter Psychostimulant-induced Behavior or Neurotoxicity

Brain Research. Jan, 2007  |  Pubmed ID: 17113054

The transcription factor NF-E2-related factor (Nrf2) regulates the induction of phase 2 detoxifying enzymes by oxidative stress, including synthesis of the catalytic subunit (xCT) of the heterodimeric cystine-glutamate exchanger (system xc-). Repeated cocaine treatment in rats causes persistent neuroadaptations in glutamate neurotransmission in the nucleus accumbens that result, in part, from reduced activity of system xc-. Since in vitro under- or over-expression of Nrf2 regulates system xc- activity and xCT content, it was hypothesized that in vivo deletion of the Nrf2 gene would: 1) decrease system xc- activity, 2) produce a behavioral phenotype resembling that elicited by chronic cocaine administration, and 3) enhance dopamine depletion after methamphetamine-induced oxidative stress. In all three experiments no genotypic difference was measured between mice sustaining homozygous Nrf2 gene deletion and wild-type littermates. Thus, while Nrf2 is a transcriptional regulator of xCT and capable of protecting cells from oxidative stress, following Nrf2 gene deletion this role can be partially compensated by other mechanisms and methamphetamine-induced oxidative stress and dopamine toxicity does not significantly involve Nrf2.

Hypothermia Modulates Circadian Clock Gene Expression in Lizard Peripheral Tissues

American Journal of Physiology. Regulatory, Integrative and Comparative Physiology. Jan, 2007  |  Pubmed ID: 16809482

The molecular mechanisms whereby the circadian clock responds to temperature changes are poorly understood. The ruin lizard Podarcis sicula has historically proven to be a valuable vertebrate model for exploring the influence of temperature on circadian physiology. It is an ectotherm that naturally experiences an impressive range of temperatures during the course of the year. However, no tools have been available to dissect the molecular basis of the clock in this organism. Here, we report the cloning of three lizard clock gene homologs (Period2, Cryptochrome1, and Clock) that have a close phylogenetic relationship with avian clock genes. These genes are expressed in many tissues and show a rhythmic expression profile at 29 degrees C in light-dark and constant darkness lighting conditions, with phases comparable to their mammalian and avian counterparts. Interestingly, we show that at low temperatures (6 degrees C), cycling clock gene expression is attenuated in peripheral clocks with a characteristic increase in basal expression levels. We speculate that this represents a conserved vertebrate clock gene response to low temperatures. Furthermore, these results bring new insight into the issue of whether circadian clock function is compatible with hypothermia.

How Reliable and Valid is the Brief Sensation Seeking Scale (BSSS-4) for Youth of Various Racial/ethnic Groups?

Addiction (Abingdon, England). Oct, 2007  |  Pubmed ID: 17850616

To assess the reliability and validity of the Brief Sensation Seeking Scale BSSS-4 by race/ethnicity.

[Percutaneous Vertebroplasty As Therapy for Vertebral Fractures: Results in a Series of Osteoporotic Patients]

Reumatismo. Jul-Sep, 2007  |  Pubmed ID: 17898880

In the recent years, percutaneous vertebroplasty is available for the treatment of the vertebral fractures, primarily to relieve pain related to the lesion. In order to evaluate the efficacy and the safety of this technique, we have treated with percutaneous vertebroplasty, using polymethylmethacrylate, 22 patients, affected by one or more vertebral fractures caused by osteoporosis. All the patients satisfied the inclusion criteria of the American College of Radiology for percutaneous vertebroplasty. These patients were compared with a control group of 23 not treated subjects with vertebral fractures, using questionnaires for assessment of pain and quality of life, drug intake, use of corset, and tolerability of the surgery. In the large majority of patients, the treatment of osteoporotic vertebral fractures with percutaneous vertebroplasty resulted in a prompt, marked and sustained relief of vertebral pain with a persistent improvement of quality of life.

[Uretero-vescical Anastomosis During Kidney Transplantation. Preliminary Results of Two Surgical Techniques]

Minerva Urologica E Nefrologica = The Italian Journal of Urology and Nephrology. Sep, 2007  |  Pubmed ID: 17912233

The most frequent urologic complications after renal transplantation involve the uretero-vescical anastomosis (leakage, stenosis, and reflux), with a frequency of 1% to 30% in different series.

Time-resolved Methods in Biophysics. 6. Time-resolved Laue Crystallography As a Tool to Investigate Photo-activated Protein Dynamics

Photochemical & Photobiological Sciences : Official Journal of the European Photochemistry Association and the European Society for Photobiology. Oct, 2007  |  Pubmed ID: 17914477

When polychromatic X-rays are shined onto crystalline material, they generate a Laue diffraction pattern. At third generation synchrotron radiation sources, a single X-ray pulse of approximately 100 ps duration is enough to produce interpretable Laue data from biomolecular crystals. Thus, by initiating biological turnover in a crystalline protein, structural changes along the reaction pathway may be filmed by ultra-fast Laue diffraction. Using laser-light as a trigger, transient species in photosensitive macromolecules can be captured at near atomic resolution with sub-nanosecond time-resolution. Such pump-probe Laue experiments have now reached an outstanding level of sophistication and have found a domain of excellence in the investigation of light-sensitive proteins undergoing cyclic photo-reactions and producing stiff crystals. The main theoretical concepts of Laue diffraction and the challenges associated with time-resolved experiments on biological crystals are recalled. The recent advances in the design of experiments are presented in terms of instrumental choices, data collection strategy and data processing, and some of the inherent difficulties of the method are highlighted. The discussion is based on the example of myoglobin, a protein that has traversed the whole history of pump-probe Laue diffraction, and for which a massive amount of data have provided considerable insight into the understanding of protein dynamics.

Successfully Maintaining Program Funding During Trying Times: Lessons from Tobacco Control Programs in Five States

Journal of Public Health Management and Practice : JPHMP. Nov-Dec, 2007  |  Pubmed ID: 17984716

Despite negative financial conditions in recent years, several states were able to successfully maintain funding for tobacco prevention and control, which provided an opportunity to understand the factors associated with success. One explanation may be the level of long-term program sustainability in some states. According to a model developed by Saint Louis University researchers, the five elements critical to tobacco control sustainability are state political and financial climate; community awareness and capacity; program structure and administration; funding stability and planning; and surveillance and evaluation. Five states (Nebraska, New York, Indiana, Virginia, and Colorado) maintained funding for their tobacco control programs. Four of these states gained additional legislative appropriations or prevented a massive reduction; Colorado used a statewide ballot initiative to increase funding. On the basis of the sustainability framework, case studies, and prior research, the major lessons learned for maintaining funding were the importance of (1) strong and experienced leadership, (2) broad and deep organizational and community ties, (3) coordinated efforts, (4) strategic use of surveillance and evaluation data, (5) active dissemination of information about program successes, and (6) policy maker champions. The sustainability framework and lessons learned may provide valuable insights for other public health programs facing funding threats.

Women's Knowledge of the Leading Causes of Cancer Death

Nicotine & Tobacco Research : Official Journal of the Society for Research on Nicotine and Tobacco. Jul, 2007  |  Pubmed ID: 17577805

This paper describes adult women's knowledge of the leading causes of cancer mortality among women. Exposure to antismoking advertisements or media messages also is examined as a potentially effective mechanism for changing inaccurate beliefs. We used data from the 2002 and 2003 American Smoking and Health Survey (ASHES), a national telephone survey of adults, to measure women's knowledge about cancer mortality. Logistic regression models were used to estimate the likelihoods of women indicating either breast or lung cancer as the leading cause of cancer mortality among women. The independent influence of individual characteristics such as race, smoking status, education, and awareness of antismoking messages or advertising on women's knowledge of cancer mortality was assessed. Overall, 66.7% of women inaccurately indicated breast cancer as the leading cause of cancer death among women, whereas 29.7% of women correctly indicated lung cancer. Black women were 43% less likely than White women to indicate lung cancer as the leading cause of cancer mortality among women. Current smokers were 35% less likely than noncurrent smokers to state that lung cancer is the leading cause of cancer mortality among women. Awareness of antismoking messages or advertisements was associated with a higher probability of correctly indicating lung cancer as the leading cause of cancer mortality among women. Our evidence suggests that antismoking media messages may help to correct inaccurate beliefs about the leading causes of cancer death among women.

A Closer Look at Smoking Among Young Adults: Where Tobacco Control Should Focus Its Attention

American Journal of Public Health. Aug, 2007  |  Pubmed ID: 17600242

We sought to fill gaps in knowledge of smoking behaviors among college-educated and non-college-educated young adults.

Antitumor Activity of PR-171, a Novel Irreversible Inhibitor of the Proteasome

Cancer Research. Jul, 2007  |  Pubmed ID: 17616698

Clinical studies with bortezomib have validated the proteasome as a therapeutic target for the treatment of multiple myeloma and non-Hodgkin's lymphoma. However, significant toxicities have restricted the intensity of bortezomib dosing. Here we describe the antitumor activity of PR-171, a novel epoxyketone-based irreversible proteasome inhibitor that is currently in clinical development. In comparison to bortezomib, PR-171 exhibits equal potency but greater selectivity for the chymotrypsin-like activity of the proteasome. In cell culture, PR-171 is more cytotoxic than bortezomib following brief treatments that mimic the in vivo pharmacokinetics of both molecules. Hematologic tumor cells exhibit the greatest sensitivity to brief exposure, whereas solid tumor cells and nontransformed cell types are less sensitive to such treatments. Cellular consequences of PR-171 treatment include the accumulation of proteasome substrates and induction of cell cycle arrest and/or apoptosis. Administration of PR-171 to animals results in the dose-dependent inhibition of the chymotrypsin-like proteasome activity in all tissues examined with the exception of the brain. PR-171 is well tolerated when administered for either 2 or 5 consecutive days at doses resulting in >80% proteasome inhibition in blood and most tissues. In human tumor xenograft models, PR-171 mediates an antitumor response that is both dose and schedule dependent. The antitumor efficacy of PR-171 delivered on 2 consecutive days is stronger than that of bortezomib administered on its clinical dosing schedule. These studies show the tolerability, efficacy, and dosing flexibility of PR-171 and provide validation for the clinical testing of PR-171 in the treatment of hematologic malignancies using dose-intensive schedules.

NAC1 Regulates the Recruitment of the Proteasome Complex into Dendritic Spines

The Journal of Neuroscience : the Official Journal of the Society for Neuroscience. Aug, 2007  |  Pubmed ID: 17699672

Coordinated proteolysis of synaptic proteins is required for synaptic plasticity, but a mechanism for recruiting the ubiquitin-proteasome system (UPS) into dendritic spines is not known. NAC1 is a cocaine-regulated transcriptional protein that was found to complex with proteins in the UPS, including cullins and Mov34. NAC1 and the proteasome were cotranslocated from the nucleus into dendritic spines in cortical neurons in response to proteasome inhibition or disinhibiting synaptic activity with bicuculline. Bicuculline also produced a progressive accumulation of the proteasome and NAC1 in the postsynaptic density. Recruitment of the proteasome into dendrites and postsynaptic density by bicuculline was prevented in neurons from mice harboring an NAC1 gene deletion or in neurons transfected with mutated NAC1 lacking the proteasome binding domain. These experiments show that NAC1 modulates the translocation of the UPS from the nucleus into dendritic spines, thereby suggesting a potential missing link in the recruitment of necessary proteolysis machinery for synaptic remodeling.

Transient Hepatic Echogenicity Difference on Contrast-enhanced Ultrasonography: Sonographic Sign and Pitfall

Journal of Ultrasound in Medicine : Official Journal of the American Institute of Ultrasound in Medicine. Mar, 2007  |  Pubmed ID: 17324983

The purpose of this study was to report and analyze a new contrast-enhanced ultrasonographic (CEUS) imaging finding, the transient hepatic echogenicity difference due to perfusion changes, using computed tomography (CT) as a reference standard.

Epithelial Misplacement in the Muscularis Propria After Biopsy of a Colonic Adenoma

Virchows Archiv : an International Journal of Pathology. May, 2007  |  Pubmed ID: 17372755

Glucocorticoids Play a Key Role in Circadian Cell Cycle Rhythms

PLoS Biology. Apr, 2007  |  Pubmed ID: 17373855

Clock output pathways play a pivotal role by relaying timing information from the circadian clock to a diversity of physiological systems. Both cell-autonomous and systemic mechanisms have been implicated as clock outputs; however, the relative importance and interplay between these mechanisms are poorly understood. The cell cycle represents a highly conserved regulatory target of the circadian timing system. Previously, we have demonstrated that in zebrafish, the circadian clock has the capacity to generate daily rhythms of S phase by a cell-autonomous mechanism in vitro. Here, by studying a panel of zebrafish mutants, we reveal that the pituitary-adrenal axis also plays an essential role in establishing these rhythms in the whole animal. Mutants with a reduction or a complete absence of corticotrope pituitary cells show attenuated cell-proliferation rhythms, whereas expression of circadian clock genes is not affected. We show that the corticotrope deficiency is associated with reduced cortisol levels, implicating glucocorticoids as a component of a systemic signaling pathway required for circadian cell cycle rhythmicity. Strikingly, high-amplitude rhythms can be rescued by exposing mutant larvae to a tonic concentration of a glucocorticoid agonist. Our work suggests that cell-autonomous clock mechanisms are not sufficient to establish circadian cell cycle rhythms at the whole-animal level. Instead, they act in concert with a systemic signaling environment of which glucocorticoids are an essential part.

Neuroglobin, Seven Years After

Cellular and Molecular Life Sciences : CMLS. May, 2007  |  Pubmed ID: 17385072

Neuroglobin is expressed in vertebrates brain and belongs to a branch of the globin family that diverged early in evolution. Sequence conservation suggests a relevant role in the nervous system, with tight structural restraints. Experiments in vivo and in vitro showed increased hypoxic stress damage upon repressing neuroglobin biosynthesis and improved recovery following overexpression. Neuroglobin shows internal heme hexacoordination, which controls oxygen affinity and kinetics. Neuroglobin concentration, oxygen affinity and enhanced autooxidation question a role in oxygen delivery; thus it was proposed that the neuroprotective effect might be due to radical scavenging or activation of protection mechanisms. Neuroglobin's structure shows a peculiar internal cavity of very large size. Binding of heme ligands is associated to a conformational change involving the heme that "slides" into the pre-existing cavity and makes the sixth coordination position available. These features may pave the way to an understanding of neuroprotection by neuroglobin.

X-ray Structure Analysis of a Metalloprotein with Enhanced Active-site Resolution Using in Situ X-ray Absorption Near Edge Structure Spectroscopy

Proceedings of the National Academy of Sciences of the United States of America. Apr, 2007  |  Pubmed ID: 17404234

X-ray absorption spectroscopy is exquisitely sensitive to the coordination geometry of an absorbing atom and therefore allows bond distances and angles of the surrounding atomic cluster to be measured with atomic resolution. By contrast, the accuracy and resolution of metalloprotein active sites obtainable from x-ray crystallography are often insufficient to analyze the electronic properties of the metals that are essential for their biological functions. Here, we demonstrate that the combination of both methods on the same metalloprotein single crystal yields a structural model of the protein with exceptional active-site resolution. To this end, we have collected an x-ray diffraction data set to 1.4-A resolution and Fe K-edge polarized x-ray absorption near edge structure (XANES) spectra on the same cyanomet sperm whale myoglobin crystal. The XANES spectra were quantitatively analyzed by using a method based on the multiple scattering approach, which yielded Fe-heme structural parameters with +/-(0.02-0.07)-A accuracy on the atomic distances and +/-7 degrees on the Fe-CN angle. These XANES-derived parameters were subsequently used as restraints in the crystal structure refinement. By combining XANES and x-ray diffraction, we have obtained an cyanomet sperm whale myoglobin structural model with a higher precision of the bond lengths and angles at the active site than would have been possible with crystallographic analysis alone.

Basic Protocols for Zebrafish Cell Lines: Maintenance and Transfection

Methods in Molecular Biology (Clifton, N.J.). 2007  |  Pubmed ID: 17417032

Cell lines derived from zebrafish embryos show great potential as cell culture tools to study the regulation and function of the vertebrate circadian clock. They exhibit directly light-entrainable rhythms of clock gene expression that can be established by simply exposing cultures to light-dark cycles. Mammalian cell lines require treatments with serum or activators of signaling pathways to initiate transient, rapidly dampening clock rhythms. Furthermore, zebrafish cells grow at room temperature, are viable for long periods at confluence, and do not require a CO2-enriched atmosphere, greatly simplifying culture conditions. Here we describe detailed methods for establishing zebrafish cell cultures as well as optimizing transient and stable transfections. These protocols have been successfully used to introduce luciferase reporter constructs into the cells and thereby monitor clock gene expression in vivo. The bioluminescence assay described here lends itself particularly well to high-throughput analysis.

Absence of Dopamine D2 Receptors Unmasks an Inhibitory Control over the Brain Circuitries Activated by Cocaine

Proceedings of the National Academy of Sciences of the United States of America. Apr, 2007  |  Pubmed ID: 17426149

Cocaine is a psychostimulant and a drug widely abused by humans. Cocaine elicits its effects primarily by blocking the activity of the dopamine (DA) transporter, leading to elevated levels of extracellular DA in areas receiving dopaminergic innervation, with the consequent activation of DA receptors. Cocaine, however, also elevates other neurotransmitter levels, leading to a general activation of interconnected brain circuitries. Studies aimed at unraveling the molecular mechanisms underlying the effects of cocaine have shown a leading role of DA D1 receptors in the cascade of cellular events elicited by this drug. In this study, we have analyzed the acute response to cocaine in animals deleted for the expression of DA D2 receptors (D2R), an inhibitor of DA signaling. Importantly, we show that although D1 receptor-mediated functions are preserved and even enhanced in D2R-/- mutants, the behavioral response to acute cocaine administration is severely altered. In addition, c-fos response to acute cocaine administration, in contrast to wild-type mice, is absent in D2R-/- mutants. Our findings show that the absence of D2R, very likely through a presynaptic mechanism, uncovers an inhibitory signaling pathway normally masked by the activity of this receptor on brain circuitries engaged by abused drugs.

Molecular Dynamics Simulation of Deoxy and Carboxy Murine Neuroglobin in Water

Biophysical Journal. Jul, 2007  |  Pubmed ID: 17468165

Globins are respiratory proteins that reversibly bind dioxygen and other small ligands at the iron of a heme prosthetic group. Hemoglobin and myoglobin are the most prominent members of this protein family. Unexpectedly a few years ago a new member was discovered and called neuroglobin (Ngb), being predominantly expressed in the brain. Ngb is a single polypeptide of 151 amino acids and despite the small sequence similarity with other globins, it displays the typical globin fold. Oxygen, nitric oxide, or carbon monoxide can displace the distal histidine which, in ferrous Ngb as well as in ferric Ngb, is bound to the iron, yielding a reversible adduct. Recent crystallographic data on carboxy Ngb show that binding of an exogenous ligand is associated to structural changes involving heme sliding and a topological reorganization of the internal cavities; in particular, the huge internal tunnel that connects the bulk with the active site, peculiar to Ngb, is heavily reorganized. We report the results of extended (90 ns) molecular dynamics simulations in water of ferrous deoxy and carboxy murine neuroglobin, which are both coordinated on the distal site, in the latter case by CO and in the former one by the distal His(64)(E7). The long timescale of the simulations allowed us to characterize the equilibrated protein dynamics and to compare protein structure and dynamical behavior coupled to the binding of an exogenous ligand. We have characterized the heme sliding motion, the topological reorganization of the internal cavities, the dynamics of the distal histidine, and particularly the conformational change of the CD loop, whose flexibility depends ligand binding.

Realization and Characterization of a Two-photon Four-qubit Linear Cluster State

Physical Review Letters. May, 2007  |  Pubmed ID: 17501550

We report on the experimental realization of a four-qubit linear cluster state via two photons entangled both in polarization and linear momentum. This state was investigated by performing tomographic measurements and by evaluating an entanglement witness. By use of this state we carried out a novel nonlocality proof, the so-called "stronger two observer all-versus-nothing" test of quantum nonlocality.

Opposing Roles for the Ventral Prefrontal Cortex and the Basolateral Amygdala on the Spontaneous Recovery of Cocaine-seeking in Rats

Psychopharmacology. Apr, 2008  |  Pubmed ID: 18066533

The neural circuitry subserving cocaine-seeking after extinction vs abstinence alone requires different constituent brain structures. Spontaneous recovery of cocaine-seeking, a model, which incorporates both extinction and abstinence, depends on an unknown neural circuit.

An Inaccurate Literature Review and Citation

Radiology. Mar, 2008  |  Pubmed ID: 18309023

The Importance of Location for Tobacco Cessation: Rural-urban Disparities in Quit Success in Underserved West Virginia Counties

The Journal of Rural Health : Official Journal of the American Rural Health Association and the National Rural Health Care Association. 2008  |  Pubmed ID: 18397443

Adults who live in rural areas of the United States have among the highest smoking rates in the country. Rural populations, including Appalachian adults, have been historically underserved by tobacco control programs and policies and little is known about their effectiveness.

An X-ray Diffraction and X-ray Absorption Spectroscopy Joint Study of Neuroglobin

Archives of Biochemistry and Biophysics. Jul, 2008  |  Pubmed ID: 18406335

Neuroglobin (Ngb) is a member of the globin family expressed in the vertebrate brain, involved in neuroprotection. A combined approach of X-ray diffraction (XRD) on single crystal and X-ray absorption spectroscopy (XAS) in solution, allows to determine the oxidation state and the structure of the Fe-heme both in the bis-histidine and the CO-bound (NgbCO) states. The overall data demonstrate that under X-ray the iron is photoreduced fairly rapidly, and that the previously reported X-ray structure of ferric Ngb [B. Vallone, K. Nienhaus, M. Brunori, G.U. Nienhaus, Proteins 56 (2004) 85-92] very likely refers to a photoreduced species indistinguishable from the dithionite reduced protein. Results from the XAS analysis of NgbCO in solution are in good agreement with XRD data on the crystal. However prolonged X-ray exposure at 15K determines CO release. This preliminary result paves the way to experiments aimed at the characterization of pentacoordinate ferrous Ngb, the only species competent in binding external ligands such as O2, CO or NO.

Structural Dynamics of Myoglobin

Methods in Enzymology. 2008  |  Pubmed ID: 18433639

Protein structure is endowed with a complex dynamic nature, which rules function and controls activity. The experimental investigations that yield information on protein dynamics are carried out in solution; however, in most cases, the determination of protein structure is carried out by crystallography that relies on the diffraction properties of a large number of molecules, in approximately the same conformation, arranged in a three-dimensional lattice. Myoglobin, maybe the most thoroughly characterized protein, has allowed the formulation of general principles in the field of protein structure-function correlation and, since the late 1990s, it has been possible to obtain directly some insight into the complex dynamic behavior of myoglobin and other proteins by Laue diffraction. This chapter describes some of the technological features involved in obtaining reliable data by time-resolved Laue crystallography, with subnanosecond time resolution. A synopsis of the more significant findings obtained by laser photolysis of myoglobin-CO crystals is also presented, emphasizing the more general aspects of dynamics relevant to the complex energy landscape of a protein.

Is Neuroglobin a Signal Transducer?

IUBMB Life. Jun, 2008  |  Pubmed ID: 18484091

Diagnostic Role of US for Biliary Atresia

Radiology. Jun, 2008  |  Pubmed ID: 18487545

Effects of Somatostatin Analog SOM230 on Cell Proliferation, Apoptosis, and Catecholamine Levels in Cultured Pheochromocytoma Cells

Journal of Molecular Endocrinology. Jun, 2008  |  Pubmed ID: 18502819

Surgery is the primary therapy for pheochromocytoma (PHEO), a catecholamine-producing tumor. Benign and malignant PHEO could develop recurrences, and the intraoperative risk of recurrent PHEO is an important unresolved issue. Non-surgical treatments of PHEO recurrence would therefore better prepare patients for reintervention as well as provide them with palliative management. We investigated the effects of the new somatostatin analog (pasireotide) SOM230 versus octreotide (OCT) in primary PHEO cell cultures (Pheo-c). Pheo-c from six benign surgical samples were set up and characterized by immunocytochemistry. Real-time PCR, using both PHEO tissues and Pheo-c, showed different levels of somatostatin receptor(1-5) mRNA expression. Cells treated with various doses of OCT or SOM230 for 48 and 72 h were analyzed to assess their effects on cell proliferation and apoptosis and catecholamine levels. Even if reduction of cell viability was observed in Pheo-c treated for 48 h with either OCT or SOM230 and this effect increased after 72 h, a more significant inhibition of cell growth as well as a significantly higher induction of apoptosis was seen in Pheo-c treated with SOM230 versus OCT. In particular, apoptosis in Pheo-c was detected after 48 h and was associated with increased expression and activation of caspase-3 and cleaved poly(ADP-ribose) polymerase. OCT 10(-6) M and SOM230 10(-7) M significantly reduced catecholamine levels. Our results indicate that while both OCT and SOM230 modulate cell growth and apoptosis and catecholamine levels in Pheo-c through specific receptors, SOM230 is more effective. This improves our knowledge on the mechanism of SOM230 action in PHEO and supports a possible therapeutic use in benign PHEO recurrence.

Active One-way Quantum Computation with Two-photon Four-qubit Cluster States

Physical Review Letters. Apr, 2008  |  Pubmed ID: 18518174

By using 2-photon 4-qubit cluster states we demonstrate deterministic one-way quantum computation in a single qubit rotation algorithm. In this operation feed-forward measurements are automatically implemented by properly choosing the measurement basis of the qubits, while Pauli error corrections are realized by using two fast driven Pockels cells. We realized also a C-NOT gate for equatorial qubits and a C-PHASE gate for a generic target qubit. Our results demonstrate that 2-photon cluster states can be used for rapid and efficient deterministic one-way quantum computing.

Moderate Zinc Restriction During Fetal and Postnatal Growth of Rats: Effects on Adult Arterial Blood Pressure and Kidney

American Journal of Physiology. Regulatory, Integrative and Comparative Physiology. Aug, 2008  |  Pubmed ID: 18525016

Intrauterine and postnatal zinc restriction may result in an adverse environment for the development of cardiovascular and renal systems. This study evaluated the effects of moderate zinc deficiency during fetal life, lactation, and/or postweaning growth on systolic blood pressure, renal function, and morphology in adult life. Female Wistar rats received low (8 ppm) or control (30 ppm) zinc diets from the beginning of pregnancy up to weaning. After weaning, male offspring of each group of mothers were fed low or control zinc diet. Systolic blood pressure, creatinine clearance, proteinuria, renal morphology, renal apoptosis. and renal oxidative stress state were evaluated after 60 days. Zinc deficiency during pre- and postweaning growth induced an increase in systolic blood pressure and a decrease in the glomerular filtration rate associated with a reduction in the number and size of nephrons. Activation of renal apoptosis, reduction in catalase activity, glutathione peroxidase activity, and glutathione levels and increase in lipid peroxidation end products could explain these morphometric changes. Zinc deficiency through pre- and postweaning growth induced more pronounced renal alteration than postweaning zinc deficiency. These animals showed signs of renal fibrosis, proteinuria, increased renal apoptosis, and higher lipid peroxidation end products. A control diet during postweaning growth did not totally overcome renal oxidative stress damage, apoptosis, and fibrosis induced by zinc deficiency before weaning. In conclusion, zinc deficiency during a critical period of renal development and maturation could induce functional and morphological alterations that result in elevated blood pressure and renal dysfunction in adult life.

Molecular Dynamics Simulation of the Neuroglobin Crystal: Comparison with the Simulation in Solution

Biophysical Journal. Nov, 2008  |  Pubmed ID: 18641072

Neuroglobin (Ngb) is a monomeric protein that, despite the small sequence similarity with other globins, displays the typical globin fold. In the absence of exogenous ligands, the ferric and the ferrous forms of Ngb are both hexacoordinated to the distal and proximal histidines. In the ferrous form, oxygen, nitric oxide or carbon monoxide can displace the distal histidine, yielding a reversible adduct. Crystallographic data show that the binding of an exogenous ligand is associated to structural changes involving heme sliding and a topological reorganization of the internal cavities. Molecular dynamics (MD) simulations in solution show that the heme oscillates between two positions, much as the ones observed in the crystal structure, although the occupancy is different. The simulations also suggest that ligand binding in solution can affect the flexibility and conformation of residues connecting the C and D helices, referred to as the CD corner, which is coupled to the configuration adopted by the distal histidine. In this study, we report the results of 30 ns MD simulations of CO-bound Ngb in the crystal. Our goal was to compare the protein dynamical behavior in the crystal with the results supplied by the previous MD simulation of CO-bound Ngb in solution and the x-ray experimental data. The results show that the different environments (crystal or solution) affect the dynamics of the heme group and of the CD corner.

Thyroid B-flow Twinkling Sign: a New Feature of Papillary Cancer

European Journal of Endocrinology / European Federation of Endocrine Societies. Oct, 2008  |  Pubmed ID: 18644823

Microcalcifications (aggregated with psammoma bodies), detected by ultrasound (US), are the most specific feature of papillary thyroid cancer (PTC). Using B-flow imaging (BFI), we identified a new sign (the twinkling sign; BFI-TS) in 'suspect' PTC nodules, which appeared to be generated by microcalcifications.

A New Marker for Diagnosis of Thyroid Papillary Cancer: B-flow Twinkling Sign

Journal of Ultrasound in Medicine : Official Journal of the American Institute of Ultrasound in Medicine. Aug, 2008  |  Pubmed ID: 18645077

The purpose of this study was to correlate the presence and patterns of distribution of B-flow imaging (BFI) twinkling signs within thyroid nodules with the histologic evidence of microcalcifications and the results of the sonographically guided fine-needle aspiration to establish their role in predicting the risk of malignancy.

Is Resistin a Link Between Highly Active Antiretroviral Therapy and Fat Redistribution in HIV-infected Children?

Journal of Endocrinological Investigation. Jul, 2008  |  Pubmed ID: 18787374

To assess the features of fat redistribution, detected by clinical and ultrasound (US) methods, and the presence of metabolic disorders in HIV-infected children undergoing antiretroviral therapy. To evaluate if serum levels of resistin, a hormone produced only by visceral adipose tissue, are a marker of fat redistribution in these patients.

Identification of West Eurasian Mitochondrial Haplogroups by MtDNA SNP Screening: Results of the 2006-2007 EDNAP Collaborative Exercise

Forensic Science International. Genetics. Jan, 2008  |  Pubmed ID: 19083791

The European DNA Profiling (EDNAP) Group performed a collaborative exercise on a mitochondrial (mt) DNA screening assay that targeted 16 nucleotide positions in the coding region and allowed for the discrimination of major west Eurasian mtDNA haplogroups. The purpose of the exercise was to evaluate the stability and reproducibility of the self-developed multiplex-PCR and multiplex-single base extension kit by blind-testing saliva and hair shaft samples provided by the organizing laboratory. The overall success rate in obtaining useful results was high given that some of the participating laboratories had no previous experience with the technology and/or mtDNA analysis. The results of this collaborative exercise stimulate the expansion of screening methods in forensic laboratories to increase efficiency and performance of mtDNA typing, and thus demonstrates that mtDNA SNP typing is a powerful tool for forensic casework analysis.

Forensic Typing of Autosomal SNPs with a 29 SNP-multiplex--results of a Collaborative EDNAP Exercise

Forensic Science International. Genetics. Jun, 2008  |  Pubmed ID: 19083818

We report the results of an inter-laboratory exercise on typing of autosomal single nucleotide polymorphisms (SNP) for forensic genetic investigations in crime cases. The European DNA Profiling Group (EDNAP), a working group under the International Society for Forensic Genetics (ISFG), organised the exercise. A total of 11 European and one US forensic genetic laboratories tested a subset of a 52 SNP-multiplex PCR kit developed by the SNPforID consortium. The 52 SNP-multiplex kit amplifies 52 DNA fragments with 52 autosomal SNP loci in one multiplex PCR. The 52 SNPs are detected in two separate single base extension (SBE) multiplex reactions with 29 and 23 SNPs, respectively, using SNaPshot kit, capillary electrophoresis and multicolour fluorescence detection. For practical reasons, only the 29 SBE multiplex reaction was carried out by the participating laboratories. A total of 11 bloodstains on FTA cards including a sample of poor quality and a negative control were sent to the laboratories together with the essential reagents for the initial multiplex PCR and the multiplex SBE reaction. The total SNP locus dropout rate was 2.8% and more than 50% of the dropouts were observed with the poor quality sample. The overall rate of discrepant SNP allele assignments was 2.0%. Two laboratories reported 60% of all the discrepancies. Two laboratories reported all 29 SNP alleles in all 10 positive samples correctly. The results of the collaborative exercise were surprisingly good and demonstrate that SNP typing with SBE, capillary electrophoresis and multicolour detection methods can be developed for forensic genetics.

Demonstration of Rapid Multiplex PCR Amplification Involving 16 Genetic Loci

Forensic Science International. Genetics. Dec, 2008  |  Pubmed ID: 19083866

Current forensic DNA typing is conducted in approximately 8-10h. Steps include DNA extraction, quantification, polymerase chain reaction (PCR) amplification of multiple short tandem repeat (STR) loci, capillary electrophoresis separation with fluorescence detection, data analysis and DNA profile interpretation. The PCR amplification portion of the workflow typically takes approximately 3h with standard thermal cycling protocols. Here we demonstrate a rapid cycling protocol that amplifies 15 STR loci and the sex-typing marker amelogenin from the Identifiler STR typing kit in less than 36 min. This rapid protocol employs commercially available polymerases and the widely used GeneAmp 9700 thermal cycler. Complete concordance of STR allele calls (for 60 samples) between the rapid and standard thermal cycling protocols were observed although there was incomplete adenylation at several of the loci examined and some PCR artifacts were detected. Using less than 750 pg of template DNA and 28 cycles, STR peaks for all loci were above a 150 relative fluorescent unit (RFU) detection threshold with fully adequate inter-locus balance and heterozygote peak height ratios of greater than 0.84.

[Angiomyofibroblastoma of the Spermatic Funiculus. A Case Report of This Lesion of the Spermatic Funiculus.]

Urologia. January-March, 2008  |  Pubmed ID: 21086377

The authors are hereby presenting a rare case of angiomyofibroblastoma of the funiculus in a 20-year-old patient, having a non-aching tumefaction at the left region of the scrotum. This tumefaction was solid and non homogeneous, both on ultrasonography and MRI, of about 5cm in diameter, fully separated from the testicle. The markers were all negative. We proceeded with surgical treatment of the neoformation. It is mandatory to include this lesion within the differential diagnosis of testicle masses. The case is presented for its absolute histopathologic rarity and for the difficulty in diagnosis.

Neuroglobin: Enzymatic Reduction and Oxygen Affinity

Biochemical and Biophysical Research Communications. Mar, 2008  |  Pubmed ID: 18198128

Neuroglobin (Ngb) is a hexacoordinate globin expressed in the nervous system of vertebrates, involved in neuroprotection. O(2) equilibrium measurements on mouse Ngb yielded significantly different P(50) values, ranging from approximately 2 torr to approximately 10 torr. By a kinetic approach minimizing the effects of protein autoxidation, we measured P(50)=2.2 torr at 20 degrees C. As predicted from the structure, O(2) binds to the Y44D Ngb mutant more quickly (k=2.2s(-1) vs 0.15s(-1)) and with slightly higher affinity (P(50)=1.3 torr) than wild-type. In addition, we introduced a novel reduction protocol for metNgb based on NADH:flavorubredoxin oxidoreductase (FlRd-red) from Escherichia coli, a candidate for the Ngb reducing activity recently identified in E. coli extracts. Interestingly, E. coli FlRd-red shares sequence similarity with the FAD-binding domain of the human apoptosis-inducing factor, a finding which may have unexpected significance with reference to the mechanism of neuroprotection by Ngb.

Voluntary or Universal HIV Testing in Italy?

Clinical Infectious Diseases : an Official Publication of the Infectious Diseases Society of America. Feb, 2008  |  Pubmed ID: 18205542

Darunavir-etravirine-raltegravir As Salvage Combination Therapy in a Heavily Pretreated HIV-infected Patient

AIDS Patient Care and STDs. Nov, 2008  |  Pubmed ID: 19025477

Cloning, Expression, Purification, Crystallization and Preliminary X-ray Crystallographic Analysis of C-12 Hydroxylase EryK from Saccharopolyspora Erythraea

Protein and Peptide Letters. 2008  |  Pubmed ID: 19075827

Erythromycin A is produced by Saccharopolyspora erythraea via a secondary metabolic pathway using several steps including glycosylations and hydroxylations of the first macrolide intermediate 6-deoxyerythronolide B. Erythromycin C-12 hydroxylase (CYP113A1), the P450 cytochrome active in the final stages of erythromycin biosynthesis, was cloned and expressed in E. coli. Different crystal forms were harvested from distinct crystallization conditions: two ligand-free forms, one substrate bound and two inhibitors-bound. All crystals belong either to the monoclinc P2(1)or to the orthorhombic P2(1)2(1)2(1) space groups, and exhibit diffraction limits ranging from 2.3 to 1.6 A. The structures will be determined by molecular replacement.

Solitary Fibrous Tumour of the Kidney with Sarcomatous Overgrowth. Case Report and Review of the Literature

APMIS : Acta Pathologica, Microbiologica, Et Immunologica Scandinavica. Nov, 2008  |  Pubmed ID: 19133003

Solitary fibrous tumour (SFT) rarely occurs in the kidney, with only one case exhibiting malignant behaviour. We report the case of a typical SFT of the kidney with sarcomatous overgrowth in a 34-year-old woman. This malignant component, grossly apparent as a nodular area arising in the context of the main tumour mass, consisted of CD34+ mitotically active atypical plump spindle- to epithelioid-shaped cells, including pleomorphic multinucleated giant cells. A novel immunohistochemical finding was diffuse and strong S-100 protein expression by sarcomatous cells. This should be kept in mind by pathologists to avoid confusion with other S-100 protein-positive malignant neoplasms.

Assessing the Impact of the National 'truth' Antismoking Campaign on Beliefs, Attitudes, and Intent to Smoke by Race/ethnicity

Ethnicity & Health. Feb, 2009  |  Pubmed ID: 19152160

To examine racial/ethnic differences in the association between exposure to the 'truth' antismoking campaign and youth's beliefs and attitudes about cigarette companies and their intent to smoke.

Cost-utility Analysis of the National Truth Campaign to Prevent Youth Smoking

American Journal of Preventive Medicine. May, 2009  |  Pubmed ID: 19211214

In 2005, the American Journal of Public Health published an article that indicated that 22% of the overall decline in youth smoking that occurred between 1999 and 2002 was directly attributable to the truth social marketing campaign launched in 2000. A remaining key question about the truth campaign is whether the economic investment in the program can be justified by the public health outcomes; that question is examined here.

Long-term Outcome of Laparoscopic Fowler-Stephens Orchiopexy in Boys with Intra-abdominal Testis

The Journal of Urology. Apr, 2009  |  Pubmed ID: 19233407

We reviewed the records of 36 pediatric patients operated on between 1990 and 1997 for high intra-abdominal testes, using the 2-step Fowler-Stephens procedure via laparoscopy.

Gestational Diabetes: is It Linked to Platelets Hyperactivity?

Platelets. Mar, 2009  |  Pubmed ID: 19235058

Proposed Bell Experiment with Genuine Energy-time Entanglement

Physical Review Letters. Jan, 2009  |  Pubmed ID: 19257405

Franson's Bell experiment with energy-time entanglement [Phys. Rev. Lett. 62, 2205 (1989)10.1103/PhysRevLett.62.2205] does not rule out all local hidden variable models. This defect can be exploited to compromise the security of Bell inequality-based quantum cryptography. We introduce a novel Bell experiment using genuine energy-time entanglement, based on a novel interferometer, which rules out all local hidden variable models. The scheme is feasible with actual technology.

Pattern of Cavities in Globins: the Case of Human Hemoglobin

Biopolymers. Dec, 2009  |  Pubmed ID: 19365817

Our aim is to shed light on the conservation of potential ligand docking sites that play an important role in ligand dynamics of globins by using the technique of filling internal cavities naturally present in hemoglobin and myoglobin with xenon atoms. In particular, we present the high resolution structures of the Xe-adduct of deoxygenated wild type human hemoglobin and a quadruple mutant (L(B10)Y and H(E7)Q in alpha and beta chains). For the sake of comparison we also determined under the same experimental conditions the xenon complex of wild type sperm whale myoglobin. The analysis revealed that the number and position of Xe binding cavities are different in the alpha and beta subunits, the latter being more similar to myoglobin. Notably, no proximal Xe docking site was detected in hemoglobin, at variance with myoglobin. The pattern of internal cavities accessibility and affinity for xenon suggests a different role for the dynamics of ligand migration in the two types of hemoglobin chains as compared to myoglobin. The number and position of hydrophobic cavities in hemoglobin are briefly discussed also in comparison with the data available for other members of the globin superfamily.

Production and Certification of NIST Standard Reference Material 2372 Human DNA Quantitation Standard

Analytical and Bioanalytical Chemistry. Jun, 2009  |  Pubmed ID: 19377837

Modern highly multiplexed short tandem repeat (STR) assays used by the forensic human-identity community require tight control of the initial amount of sample DNA amplified in the polymerase chain reaction (PCR) process. This, in turn, requires the ability to reproducibly measure the concentration of human DNA, [DNA], in a sample extract. Quantitative PCR (qPCR) techniques can determine the number of intact stretches of DNA of specified nucleotide sequence in an extremely small sample; however, these assays must be calibrated with DNA extracts of well-characterized and stable composition. By 2004, studies coordinated by or reported to the National Institute of Standards and Technology (NIST) indicated that a well-characterized, stable human DNA quantitation certified reference material (CRM) could help the forensic community reduce within- and among-laboratory quantitation variability. To ensure that the stability of such a quantitation standard can be monitored and that, if and when required, equivalent replacement materials can be prepared, a measurement of some stable quantity directly related to [DNA] is required. Using a long-established conventional relationship linking optical density (properly designated as decadic attenuance) at 260 nm with [DNA] in aqueous solution, NIST Standard Reference Material (SRM) 2372 Human DNA Quantitation Standard was issued in October 2007. This SRM consists of three quite different DNA extracts: a single-source male, a multiple-source female, and a mixture of male and female sources. All three SRM components have very similar optical densities, and thus very similar conventional [DNA]. The materials perform very similarly in several widely used gender-neutral assays, demonstrating that the combination of appropriate preparation methods and metrologically sound spectrophotometric measurements enables the preparation and certification of quantitation [DNA] standards that are both maintainable and of practical utility.

Preparation of Activated Carbons from Coffee Husks Utilizing FeCl3 and ZnCl2 As Activating Agents

Journal of Hazardous Materials. Jun, 2009  |  Pubmed ID: 18996644

Ferric chloride was used as a new activating agent, to obtain activated carbons (AC) from agro industrial waste (coffee husks). This material was compared with two samples from the same raw material: one of them activated by using the classical activating agent, zinc chloride, and the other, activated with a mixture of the two mentioned activating agents in the same mass proportion. The carbonaceous materials obtained after the activation process showed high specific surface areas (BET), with values higher than 900 m(2)g(-1). It is interesting to observe that the activation with FeCl(3) produces smaller pores compared to the activation with ZnCl(2). An important fact to emphasize in the use of FeCl(3) as activating agent is the activation temperature at 280 degrees C, which is clearly below to the temperature commonly employed for chemical or physical activation, as described in the bibliography. All the studied materials showed different behaviors in the adsorption of methylene blue dye and phenol from aqueous solutions.

Is Socioeconomic Status Associated with Awareness of and Receptivity to the Truth Campaign?

Drug and Alcohol Dependence. Oct, 2009  |  Pubmed ID: 19442459

The truth campaign is credited with preventing approximately 450,000 youth from starting to smoke, from 2000 through 2004 [Farrelly, M.C., Nonnemaker, J., Davis, K.C., Hussin, A., 2009. The Influence of the National Truth Campaign on Smoking Initiation. Am. J. Prev. Med. February 9 [Epub ahead of print]]. However, no research has been conducted to determine whether lower socioeconomic status (SES) youth benefit equally from the campaign. This study examines whether socioeconomic status is associated with awareness of and receptivity to the truth campaign among youth ages 12-17.

Occurrence of Ankylosing Spondylitis and Multiple Sclerosis-like Syndrome in a HLA-B27 Positive Patient

Neurological Sciences : Official Journal of the Italian Neurological Society and of the Italian Society of Clinical Neurophysiology. May, 2009  |  Pubmed ID: 19444380

Occurrence of multiple sclerosis (MS) in patients with ankylosing spondylitis (AS) has been reported in isolated cases. We describe a white 33-year-old male with a definite familial HLAB27 positive AS and MS-like syndrome. The patient developed acute onset of gait difficulty, postural unsteadiness, dysarthria and right side weakness that resolved within 1 month; after 6 months he presented right-sided face sensory loss, disappeared after 2 weeks. Brain and cervical MRI was performed twice and showed disseminated lesions in space (multiple foci of increased signal intensity in the periventricular white matter, in the corpus callosum, in the hypothalamus, in the brainstem and in the cervical spinal cord) and in time (a new enhancing lesion >3 months after the onset of the clinical event). Visual evoked potentials were markedly altered. Cerebrospinal fluid examination was negative for intrathecal production of oligoclonal bands. Differential diagnosis was considered and other pathologies were excluded.

Neuroradiological Follow-up of the Growth of Papillary Tumor of the Pineal Region: a Case Report

Journal of Neuro-oncology. Dec, 2009  |  Pubmed ID: 19517065

Papillary tumor of the pineal region (PTPR) is a recently described distinct clinicopathological entity. The purpose of this case report is to increase the knowledge of its neuroradiological findings and natural history by describing the long-term clinical and neuroradiological follow-up of a PTPR occurring in a 56-year-old Italian male patient. At magnetic resonance imaging (MRI) obtained at diagnosis, the lesion showed a subtle high signal intensity on T1-weighted imaging. Twenty-nine months later, MRI showed clearcut enlargement of the lesion, which had only a small area of high signal intensity on T1-weighted images, and a minimum apparent diffusion coefficient of 0.854 x 10(-3) mm(2)/s. Treatment included surgery followed by irradiation. Three-month MRI follow-up did not show disease relapse.

Multipath Entanglement of Two Photons

Physical Review Letters. Apr, 2009  |  Pubmed ID: 19518632

We present a novel optical device based on an integrated system of microlenses and single-mode optical fibers. It allows us to collect and direct into many modes two photons generated by spontaneous parametric down-conversion. By this device multiqubit entangled states and/or multilevel qudit states of two photons, encoded in the longitudinal momentum degree of freedom, are created. The multipath photon entanglement realized by this device is expected to find important applications in modern quantum information technology.

Cytoreductive Surgery (cs) and Hyperthermic Intraperitoneal Chemotherapy (hipec) in Treatment of Peritoneal Surface Malignances: Report of a Phase II Clinical Study

In Vivo (Athens, Greece). Jul-Aug, 2009  |  Pubmed ID: 19567402

Peritoneal surface malignancy is the expression of a spectrum of disease involving the peritoneum primary or secondary to gastrointestinal and gynecological neoplasms. Even if intraperitoneal therapy has now been demonstrated in multiple randomized trials to improve the outcome of chemotherapy for patients with optimally debulked or small volume ovarian carcinoma, it is believed that peritoneal carcinomatosis is considered an advanced stage of disease; for this reason, it is treated with systemic chemotherapy and surgery plays only a palliative role (1). In the last twenty years, some centres have developed surgical treatment of peritoneal carcinomatosis that involves aggressive cytoreductive surgery associated with hyperthermic intraperitoneal chemotherapy. This treatment has improved and prolonged survival, despite the associated high morbidities and mortalities (3-14).

Unintended Consequences of Tobacco Policies: Implications for Public Health Practice

American Journal of Preventive Medicine. Aug, 2009  |  Pubmed ID: 19591760

Investigating the Structural Plasticity of a Cytochrome P450: Three-dimensional Structures of P450 EryK and Binding to Its Physiological Substrate

The Journal of Biological Chemistry. Oct, 2009  |  Pubmed ID: 19625248

Cytochrome P450s are heme-containing proteins that catalyze the oxidative metabolism of many physiological endogenous compounds. Because of their unique oxygen chemistry and their key role in drug and xenobiotic metabolism, particular attention has been devoted in elucidating their mechanism of substrate recognition. In this work, we analyzed the three-dimensional structures of a monomeric cytochrome P450 from Saccharopolyspora erythraea, commonly called EryK, and the binding kinetics to its physiological ligand, erythromycin D. Three different structures of EryK were obtained: two ligand-free forms and one in complex with its substrate. Analysis of the substrate-bound structure revealed the key structural determinants involved in substrate recognition and selectivity. Interestingly, the ligand-free structures of EryK suggested that the protein may explore an open and a closed conformation in the absence of substrate. In an effort to validate this hypothesis and to investigate the energetics between such alternative conformations, we performed stopped-flow absorbance experiments. Data demonstrated that EryK binds erythromycin D via a mechanism involving at least two steps. Contrary to previously characterized cytochrome P450s, analysis of double jump mixing experiments confirmed that this complex scenario arises from a pre-existing equilibrium between the open and closed subpopulations of EryK, rather than from an induced-fit type mechanism.

A 26plex Autosomal STR Assay to Aid Human Identity Testing*

Journal of Forensic Sciences. Sep, 2009  |  Pubmed ID: 19627417

A short tandem repeat multiplex assay has been successfully developed with 25 autosomal loci plus the sex-typing locus amelogenin for a total of 26 amplified products in a single reaction. Primers for the loci were designed so that all of the amplicons present were distributed from 65 base pairs (bp) to less than 400 bp within a five-dye chemistry design with the fifth dye reserved for the sizing standard. A multiplex design strategy was developed to overcome challenges encountered in creating this assay. The limits of the multiplex were tested, resulting in the successful amplification of a wide range of genomic DNA sample concentrations from 2 ng to as low as 100 pg with 30 cycles of PCR. The 26plex has the potential to benefit the forensic community for reference sample testing and complex relationship evaluation.

Prevalence, Characteristics and Severity of Non-alcoholic Fatty Liver Disease in Patients with Chronic Plaque Psoriasis

Journal of Hepatology. Oct, 2009  |  Pubmed ID: 19664838

The association between NAFLD and psoriasis has never been explored in prospective epidemiological studies. The aim of this 2-phase study was to study the clinical features of NAFLD in patients with psoriasis.

The Structure of Neuroglobin at High Xe and Kr Pressure Reveals Partial Conservation of Globin Internal Cavities

Biophysical Journal. Sep, 2009  |  Pubmed ID: 19751675

Neuroglobin (Ngb) is a hexacoordinate globin expressed in the brain of vertebrates. Ferrous Ngb binds dioxygen with high affinity and the O(2) adduct is able to scavenge NO. Convincing in vitro and in vivo data indicate that Ngb is involved in neuroprotection during hypoxia and ischemia. The 3D structure of Ngb reveals the presence of a wide internal cavity connecting its heme active site with the bulk. To explore the role of this "tunnel" in the control of ligand binding, we determined the structure of metNgb and NgbCO equilibrated with Xe or Kr. We show four docking sites for Xe (only two for Kr); two of the four Xe sites are within the large cavity. They are only partially conserved in globins, since the two proximal Xe sites identified in myoglobin (Xe1 and Xe2) are absent in Ngb, as well as in cytoglobin. The Xe docking sites in Ngb map a pathway within the protein matrix, leading to the heme, which becomes more accessible in the ligand-bound species. This may be of significance in connection with the redox chemistry that may be the primary function of this hexacoordinate globin.

Can Hepatocellular Cancer Screening Increase the Proportion of Long-term Survivors?

Hepato-gastroenterology. Jul-Aug, 2009  |  Pubmed ID: 19760960

Historically, only 10% of hepatocellular cancer (HCC) patients are diagnosed with early stage, potentially curable disease. We prospectively screened chronic hepatitis virus-infected patients to determine 1) the proportion diagnosed with potentially curable HCC, and 2) survival following curative therapy.

SNP Genotyping Using Multiplex Single Base Primer Extension Assays

Methods in Molecular Biology (Clifton, N.J.). 2009  |  Pubmed ID: 19768606

Single nucleotide polymorphisms (SNPs) are the most common form of polymorphisms present in the human genome. The single base primer extension (SBE) method is an effective and sensitive tool that can type over 30 known loci scattered throughout an organism's genome in a single reaction. It allows the typing of tetra-allelic SNPs and has been adapted to a broad range of analytical necessities: single-cell analysis, molecular diagnosis of monogenic diseases, forensic mitochondrial DNA analysis on highly degraded human remains, and high-throughput SNP screening for population studies. Every SBE-based assay will need customized optimization efforts that are generally proportional to the number of desired SNPs typed in a single reaction. This chapter offers a detailed outline on which to base the design and optimization of any multiplex SBE assay that can then be tailored to the analytical conditions that characterize each specific application.

Safety and Efficacy of Pegylated Liposomal Doxorubicin in HIV-associated Kaposi's Sarcoma

Biologics : Targets & Therapy. 2009  |  Pubmed ID: 19774206

Kaposi's sarcoma is a vascular tumor linked to the presence of Kaposi's sarcoma-associated herpesvirus (human herpesvirus-8) and the incidence of which has increased considerably the world over after the onset of the human immunodeficiency virus (HIV) pandemic. Antiretroviral therapy combined with cytotoxic agents has been established as the treatment of choice in the past 10 years. Among chemotherapeutic agents, pegylated liposomal doxorubicin has become the preferred one for patients with HIV-associated Kaposi's sarcoma in Western countries. The drug in this formulation localizes better to the tumor and has higher efficacy. Skin toxicity, mucositis, and leukopenia/neutropenia are the main side effects. Hepatotoxicity and mild cardiotoxicity are observed less frequently. Pegylated liposomal doxorubicin impacts favorably on quality of life. Although cost effective in Western countries, the drug is less so in developing countries.

Failure of Apoptosis-inducing Factor to Act As Neuroglobin Reductase

Biochemical and Biophysical Research Communications. Dec, 2009  |  Pubmed ID: 19782043

Neuroglobin (Ngb) is a hexacoordinate globin expressed in the nervous system of vertebrates, where it protects neurons against hypoxia. Ferrous Ngb has been proposed to favor cell survival by scavenging NO and/or reducing cytochrome c released into the cytosol during hypoxic stress. Both catalytic functions require an as yet unidentified Ngb-reductase activity. Such an activity was detected both in tissue homogenates of human brain and liver and in Escherichia coli extracts. Since NADH:flavorubredoxin oxidoreductase from E. coli, that was shown to reduce ferric Ngb, shares sequence similarity with the human apoptosis-inducing factor (AIF), AIF has been proposed by us as a candidate Ngb reductase. In this study, we tested this hypothesis and show that the Ngb-reductase activity of recombinant human AIF is negligible and hence incompatible with such a physiological function.

Increasing Youths' Exposure to a Tobacco Prevention Media Campaign in Rural and Low-population-density Communities

American Journal of Public Health. Dec, 2009  |  Pubmed ID: 19833994

We examined the effectiveness of a program to increase exposure to national "truth" tobacco countermarketing messages among youths in rural and low-population-density communities.

Light Directs Zebrafish Period2 Expression Via Conserved D and E Boxes

PLoS Biology. Oct, 2009  |  Pubmed ID: 19859524

For most species, light represents the principal environmental signal for entraining the endogenous circadian clock. The zebrafish is a fascinating vertebrate model for studying this process since unlike mammals, direct exposure of most of its tissues to light leads to local clock entrainment. Importantly, light induces the expression of a set of genes including certain clock genes in most zebrafish cell types in vivo and in vitro. However, the mechanism linking light to gene expression remains poorly understood. To elucidate this key mechanism, here we focus on how light regulates transcription of the zebrafish period2 (per2) gene. Using transgenic fish and stably transfected cell line-based assays, we define a Light Responsive Module (LRM) within the per2 promoter. The LRM lies proximal to the transcription start site and is both necessary and sufficient for light-driven gene expression and also for a light-dependent circadian clock regulation. Curiously, the LRM sequence is strongly conserved in other vertebrate per2 genes, even in species lacking directly light-sensitive peripheral clocks. Furthermore, we reveal that the human LRM can substitute for the zebrafish LRM to confer light-regulated transcription in zebrafish cells. The LRM contains E- and D-box elements that are critical for its function. While the E-box directs circadian clock regulation by mediating BMAL/CLOCK activity, the D-box confers light-driven expression. The zebrafish homolog of the thyrotroph embryonic factor binds efficiently to the LRM D-box and transactivates expression. We demonstrate that tef mRNA levels are light inducible and that knock-down of tef expression attenuates light-driven transcription from the per2 promoter in vivo. Together, our results support a model where a light-dependent crosstalk between E- and D-box binding factors is a central determinant of per2 expression. These findings extend the general understanding of the mechanism whereby the clock is entrained by light and how the regulation of clock gene expression by light has evolved in vertebrates.

Glyphosate Resistance by Engineering the Flavoenzyme Glycine Oxidase

The Journal of Biological Chemistry. Dec, 2009  |  Pubmed ID: 19864430

Glycine oxidase from Bacillus subtilis is a homotetrameric flavoprotein of great potential biotechnological use because it catalyzes the oxidative deamination of various amines and d-isomer of amino acids to yield the corresponding alpha-keto acids, ammonia/amine, and hydrogen peroxide. Glyphosate (N-phosphonomethylglycine), a broad spectrum herbicide, is an interesting synthetic amino acid: this compound inhibits 5-enolpyruvylshikimate-3-phosphate synthase in the shikimate pathway, which is essential for the biosynthesis of aromatic amino acids in plants and certain bacteria. In recent years, transgenic crops resistant to glyphosate were mainly generated by overproducing the plant enzyme or by introducing a 5-enolpyruvylshikimate-3-phosphate synthase insensitive to this herbicide. In this work, we propose that the enzymatic oxidation of glyphosate could be an effective alternative to this important biotechnological process. To reach this goal, we used a rational design approach (together with site saturation mutagenesis) to generate a glycine oxidase variant more active on glyphosate than on the physiological substrate glycine. The glycine oxidase containing three point mutations (G51S/A54R/H244A) reaches an up to a 210-fold increase in catalytic efficiency and a 15,000-fold increase in the specificity constant (the k(cat)/K(m) ratio between glyphosate and glycine) as compared with wild-type glycine oxidase. The inspection of its three-dimensional structure shows that the alpha2-alpha3 loop (comprising residues 50-60 and containing two of the mutated residues) assumes a novel conformation and that the newly introduced residue Arg(54) could be the key residue in stabilizing glyphosate binding and destabilizing glycine positioning in the binding site, thus increasing efficiency on the herbicide.

A Phase 1 Dose Escalation Study of the Safety and Pharmacokinetics of the Novel Proteasome Inhibitor Carfilzomib (PR-171) in Patients with Hematologic Malignancies

Clinical Cancer Research : an Official Journal of the American Association for Cancer Research. Nov, 2009  |  Pubmed ID: 19903785

Carfilzomib (formerly PR-171) is a novel proteasome inhibitor of the epoxyketone class that is selective and structurally distinct from bortezomib. Proteasome inhibition by carfilzomib is mechanistically irreversible. Consequently, proteasome inhibition is more sustained with carfilzomib than with bortezomib.

Experimental Entanglement and Nonlocality of a Two-photon Six-qubit Cluster State

Physical Review Letters. Oct, 2009  |  Pubmed ID: 19905673

We create a six-qubit linear cluster state by transforming a two-photon hyperentangled state in which three qubits are encoded in each particle, one in the polarization and two in the linear momentum degrees of freedom. For this state, we demonstrate genuine six-qubit entanglement, persistency of entanglement against the loss of qubits, and higher violation than in previous experiments on Bell inequalities of the Mermin type.

Transplacental Transfer of Antiretroviral Drugs and Newborn Birth Weight in HIV-infected Pregnant Women

Current HIV Research. Nov, 2009  |  Pubmed ID: 19929798

Although it is well known that antiretroviral drugs (ARVs) across the placenta in different extents, few data are available concerning the impact of the transplacental passage of ARVs on newborn outcome. The aim of this study is to evaluate the transplacental diffusion of ARVs and the clinical assessment of the newborn. Mother and cord lopinavir, nelfinavir, atazanavir and nevirapine plasma levels were determined by high-performance liquid chromatography. Newborn gestational age, weight, and Apgar score were recorded. Cord-to-mother ratio (C:M) was calculated to estimate the placental passage of ARVs. Preterm birth was defined as delivery at <37 weeks of gestation and low birth weight was defined as a birth weight of <2500g. Twenty-six HIV-infected pregnant women were enrolled. Nevirapine presented the highest C:M ratio (0.60 +/- 0.19), the C:M ratio of nelfinavir and atazanavir was 0.37 +/- 0.38 and 0.20 +/- 0.14, respectively. The lopinavir level in the cord was undetectable. The observed prevalence rate of neonatal low birth weight and preterm delivery was 19,2% (n = 5) and 15.4% (n = 4), respectively. A significant linear regression analysis was reported between the C:M ratio and newborn birth weight (p = 0.01). Although the role of highly active antiretroviral therapy (HAART) in preventing mother-to-child transmission is indisputable, these data indicate a pharmacological rationale to the association between birth weight and highly active antiretroviral therapy during pregnancy.

Inclusion of Chiropractic Care in Multidisciplinary Management of a Child with Prader-Willi Syndrome: a Case Report

Journal of Chiropractic Medicine. Dec, 2009  |  Pubmed ID: 19948310

The purpose of this paper is to present a case of a child with Prader-Willi syndrome and the observed improvement in the degree of scoliosis, immune function, and behavior documented during the course of her treatment.

A Dramatic Storm of Idiopathic Ventricular Fibrillation

Clinical Research in Cardiology : Official Journal of the German Cardiac Society. Jan, 2009  |  Pubmed ID: 19096890

Fetal Upper Limb Amelia with Increased Nuchal Translucency

Congenital Anomalies. Sep, 2009  |  Pubmed ID: 20002906

Total Occlusion of a Conus Medullaris Pial Arteriovenous Malformation Obtained with One Session of Superselective Embolization

Journal of Neurosurgical Sciences. Sep, 2009  |  Pubmed ID: 20075824

The authors report about a case of the endovascular treatment of a pial arteriovenous malformation (AVM). The lesion was located on the conus medullaris. This injury is a rare spinal AVM. The diagnostic management and surgical treatment was chosen with a collaboration between neurosurgeons and neuroradiologists. The diagnostic management was based on clinical validation and magnetic resonance with angiographic technique as a gold standard. With regard to the surgical treatment of spinal AVM, endovascular and radiotherapy is a decision which should be taken multidisciplinarily. The treatment is crucial in resolving this lesion. The authors describe the case of a 38-year-old girl with clinical findings of progressive radiculomedullary ischemic process caused by the presence of spinal AVM. The angiographic images showed a pial AVM of the conus medullaris fed by an anterior radiculomedullary artery (Adamckiewiz artery) originated from a left T11 dorsospinal artery and by a posterior radiculopial artery originated from the left L1 artery. The draining veins were posterior pial veins, and accessory anterior subpial veins. Even if the first treatment of a pial arteriovenous malformation (AVM) of conus medullaris can be the surgical treatment for posterior localization, a neurointerventional angiographic and modern materials make it possible to reach pial AVMS of conus medullaris avoiding surgery. The authors describe a successful treatment of conus medullaris arteriovenous malformation with a one session of superselective embolization.

Monitoring the Tobacco Use Epidemic I. Overview: Optimizing Measurement to Facilitate Change

Preventive Medicine. Jan, 2009  |  Pubmed ID: 18809429

This Overview paper (I of V) summarizes research work to date on monitoring the tobacco use epidemic, discusses the recommendations made at the November, 2002 National Tobacco Monitoring, Research and Evaluation Workshop sponsored by the U.S. National Cancer Institute (NCI), Centers for Disease Control and Prevention (CDC), the American Legacy Foundation, and the Robert Wood Johnson Foundation on the topic of tobacco surveillance and evaluation, and discusses the current state of affairs.

Neuronal Pentraxins Modulate Cocaine-induced Neuroadaptations

The Journal of Pharmacology and Experimental Therapeutics. Jan, 2009  |  Pubmed ID: 18840757

Neuronal pentraxins (NPs) function in the extracellular matrix to bind alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors. Three NPs have been described, neuronal activity-regulated pentraxin (Narp), which is regulated as an immediate early gene, NP1, and neuronal pentraxin receptor (NPR). Narp and NP1 enhance synaptogenesis and glutamate signaling by clustering AMPA receptors, whereas NPR contributes to removing AMPA receptors during group I metabotropic glutamate receptor-dependent long-term depression. Here, we examine mice with genetic deletions [knockout (KO)] of each NP to assess their contributions to cocaine-induced neuroplasticity. Consistent with a shared AMPA receptor clustering function for Narp and NP1, deletion of either NP caused similar behavioral alterations. Thus, although both Narp and NP1 deletion promoted cocaine-induced place preference, NPR deletion was without effect. In addition, although Narp and NP1 KO showed reduced time in the center of a novel environment, NPR KO mice spent more time in the center. Finally, although Narp and NP1 KO mice showed blunted locomotion after AMPA microinjection into the accumbens 3 weeks after discontinuing repeated cocaine injections, the AMPA response was augmented in NPR KO. Likewise, endogenous glutamate release elicited less motor activity in Narp KO mice. Consistent with reduced AMPA responsiveness after chronic cocaine in Narp KO mice, glutamate receptor 1 was reduced in the PSD fraction of Narp KO mice withdrawn from cocaine. These data indicate that NPs differentially contribute to cocaine-induced plasticity in a manner that parallels their actions in synaptic plasticity.

Monoblastic Leukaemia/sarcoma of the Skin with an Indeterminate Cell Immunophenotype: Cyto-haematological Correlation

Cytopathology : Official Journal of the British Society for Clinical Cytology. Dec, 2010  |  Pubmed ID: 20109163

Lymph Nodes and Pathogenesis of Infection with HIV-1

The Lancet Infectious Diseases. Feb, 2010  |  Pubmed ID: 20113975

Nephron-sparing Surgery: Our Experience in Open and Laparoscopic Approach in 254 Cases

Archivos Españoles De Urología. Jan-Feb, 2010  |  Pubmed ID: 20157221

The majority of renal cell carcinomas are now incidentally detected as small renal masses in asymptomatic patients due to the widespread use of ultrasound and new improved noninvasive abdominal imaging modalities. Nephron-sparing surgery is the treatment of choice for patients with small renal masses in presence of normal contralateral kidney or in presence of an anatomic or functional solitary kidney. We reviewed retrospectively our experience in open and laparoscopic partial nephrectomy.

The Impact of EX: Results from a Pilot Smoking-cessation Media Campaign

American Journal of Preventive Medicine. Mar, 2010  |  Pubmed ID: 20176302

Mass media campaigns can be an effective strategy to increase quitting activity among smokers, particularly when aired in the context of other anti-tobacco efforts.

Preoperative Coronary Risk Assessment with Dual-source CT in Patients Undergoing Noncoronary Cardiac Surgery

La Radiologia Medica. Oct, 2010  |  Pubmed ID: 20221708

The aim of our work was to assess the role of dual-source computed tomography (DSCT) in the preoperative evaluation of coronary artery disease in patients scheduled for noncoronary cardiac surgery.

[Delirium in Elderly Inpatients. An 18 Month Follow-up]

Medicina. 2010  |  Pubmed ID: 20228018

Delirium usually hardens care during hospitalization and increases morbidity during hospital stay and after discharge. The objective of this study was to describe the prevalence of delirium in elderly inpatients in a Buenos Aires hospital, its morbidity and mortality during hospital stay and the next 18 month follow-up. Patients aged 70 or older admitted to internal medicine unit between September 2005 and May 2006 were enrolled. Delirium was assessed with the Spanish version of Confusion Assessment Method. Demographic data, cause of admition and length of stay, destination after discharge and mortality were registered. A new evaluation was made 18 months after discharge. We evaluated 194 patients and 74 were excluded. Of the 120 included, 52 (43.3%) presented delirium. We found significant differences between patients with and without delirium in previous placement in nursing home (17.3% vs. 1.5%; p < 0.002), dementia (40.4% vs. 8.8%; p < 0.001), median activity of daily living (5 vs. 6; p < 0.001), length-of-stay (7 vs. 5; p = 0.04) and mortality rate (21.2% vs. 1.5%; p < 0.001). Evaluation 18 months later showed differences between patients with and without delirium in median of activity of daily living (1/6 vs. 5/6), patients living in nursing homes (27.5% vs. 7.9%), estimated survival 35.3% (CI 95%: 24-49%) at day 569 and 49% (CI 95%: 32.9-65.4%) at day 644. The difference between survival curves was statistically significant (p = 0.027). Delirium increases morbidity and mortality during hospital stay. Elderly with delirium are at risk of worsening disability and of becoming dependent after discharge and it is a risk factor for higher mortality during the following months after discharge.

Camel No. 9 Cigarette-marketing Campaign Targeted Young Teenage Girls

Pediatrics. Apr, 2010  |  Pubmed ID: 20231181

The 1998 Master Settlement Agreement (MSA) restricted tobacco industry advertising practices that targeted teens.

Prior Mycobacterium Avium Complex Infection is Linked to Immunological Nonresponsiveness in HIV-infected Patients on Highly Active Antiretroviral Therapy

HIV Medicine. Sep, 2010  |  Pubmed ID: 20236362

Use of Darunavir/ritonavir Once Daily in Treatment-naive Pregnant Woman: Pharmacokinetics, Compartmental Exposure, Efficacy and Safety

AIDS (London, England). Apr, 2010  |  Pubmed ID: 20386380

Sonography for Locoregional Staging and Follow-up of Cutaneous Melanoma: How We Do It

Journal of Ultrasound in Medicine : Official Journal of the American Institute of Ultrasound in Medicine. May, 2010  |  Pubmed ID: 20427792

Sonography is being used with increasing frequency in the assessment of locoregional tumor spread in patients with melanoma. Nevertheless, to maximize its practical impact, sonography should be performed with state-of-the-art equipment, by specifically trained operators, and using a careful exploration technique and well-defined diagnostic criteria. In this "how I do it"-type article, we illustrate our practical approach to sonography of cutaneous melanoma.

Determination of Styrene Content in Gorgonzola PDO Cheese by Headspace Solid Phase Micro-extraction (HS-SPME) and Gas-chromatography Mass-spectrometry (GC-MS)

Veterinary Research Communications. Jun, 2010  |  Pubmed ID: 20461461

Control of the composition of products that are intended for use as packaging material is essential, particularly when these products come into direct contact with food. It is well known that plastics are not inert and that their residual monomers, starting substances, and additives are able to migrate into the food they contact. Among plastics, styrene is a common compound found in many plastic containers that can also be produced by the oxidation of Penicillium roqueforti used in gorgonzola Protected Denomination of Origin cheese manufacturing. Therefore, solid-phase microextraction combined with gas chromatography/mass spectrometry was applied in the present work to determine the styrene content in packaged and unpackaged gorgonzola cheese samples to understand styrene migration phenomena from plastic containers.

Chiropractic Approach to the Management of Children

Chiropractic & Osteopathy. 2010  |  Pubmed ID: 20525200

Chiropractic (Greek: done by hand) is a health care profession concerned with the diagnosis, treatment and prevention of disorders of the neuromusculoskeletal system and the effects of these disorders on general health. There is an emphasis on manual techniques, including joint adjustment and/or manipulation, with a particular focus on joint subluxation (World Health Organization 2005) or mechanical lesion and restoring function. The chiropractor's role in wellness care, prevention and treatment of injury or illness is based on education in anatomy and physiology, nutrition, exercise and healthy lifestyle counseling as well as referral to other health practitioners. Depending on education, geographic location, scope of practice, as well as consumer preference, chiropractors may assume the role of primary care for families who are pursuing a more natural and holistic approach to health care for their families.

Neuroimaging of Transient Ischemic Attack

Archives of Neurology. Jun, 2010  |  Pubmed ID: 20558403

T-cell Infiltration in Autosomal Dominant Neovascular Inflammatory Vitreoretinopathy

Molecular Vision. 2010  |  Pubmed ID: 20596252

Autosomal dominant neovascular inflammatory vitreoretinopathy (ADNIV) is a familial blinding disease of unknown pathophysiology. The eyes and sera from patients with ADNIV were studied to understand the immune response in this condition.

Dual Energy CT (DECT) of the Liver: Conventional Versus Virtual Unenhanced Images

European Radiology. Dec, 2010  |  Pubmed ID: 20623126

To compare image quality and noise of conventional unenhanced (CU) and virtual unenhanced (VU) images in patients who underwent hepatic dual energy computed tomography (DECT) and to assess potential radiation dose reduction.

The Role of Reported Tobacco-specific Media Exposure on Adult Attitudes Towards Proposed Policies to Limit the Portrayal of Smoking in Movies

Tobacco Control. Jun, 2010  |  Pubmed ID: 20008152

To assess the relative, independent contribution of reported tobacco-specific media exposure (pro-tobacco advertising, anti-tobacco advertising, and news coverage of tobacco issues) to US adults' support for policy efforts that aim to regulate the portrayal of smoking in movies.

The Role of Tobacco-specific Media Exposure, Knowledge, and Smoking Status on Selected Attitudes Toward Tobacco Control

Nicotine & Tobacco Research : Official Journal of the Society for Research on Nicotine and Tobacco. Feb, 2010  |  Pubmed ID: 20018944

In August 2007, the President's Cancer Panel urged the leadership of the nation to "summon the political will to address the public health crisis caused by tobacco use" (President's Cancer Panel, N, 2007, Promoting healthy lifestyles: Policy, program, and personal recommendations for reducing cancer risk. http://deainfo.nci.nih.gov/advisory/pcp/pcp07rpt/pcp07rpt.pdf). While some research has examined predictors of public support for tobacco control measures, little research has examined modifiable factors that may influence public attitudes toward tobacco control.

Inferring Continental Ancestry of Argentineans from Autosomal, Y-chromosomal and Mitochondrial DNA

Annals of Human Genetics. Jan, 2010  |  Pubmed ID: 20059473

We investigated the bio-geographic ancestry of Argentineans, and quantified their genetic admixture, analyzing 246 unrelated male individuals from eight provinces of three Argentinean regions using ancestry-sensitive DNA markers (ASDM) from autosomal, Y and mitochondrial chromosomes. Our results demonstrate that European, Native American and African ancestry components were detectable in the contemporary Argentineans, the amounts depending on the genetic system applied, exhibiting large inter-individual heterogeneity. Argentineans carried a large fraction of European genetic heritage in their Y-chromosomal (94.1%) and autosomal (78.5%) DNA, but their mitochondrial gene pool is mostly of Native American ancestry (53.7%); instead, African heritage was small in all three genetic systems (<4%). Population substructure in Argentina considering the eight sampled provinces was very small based on autosomal (0.92% of total variation was between provincial groups, p = 0.005) and mtDNA (1.77%, p = 0.005) data (none with NRY data), and all three genetic systems revealed no substructure when clustering the provinces into the three geographic regions to which they belong. The complex genetic ancestry picture detected in Argentineans underscores the need to apply ASDM from all three genetic systems to infer geographic origins and genetic admixture. This applies to all worldwide areas where people with different continental ancestry live geographically close together.

Azole Drugs Trap Cytochrome P450 EryK in Alternative Conformational States

Biochemistry. Nov, 2010  |  Pubmed ID: 20845962

EryK is a bacterial cytochrome P450 that catalyzes the last hydroxylation occurring during the biosynthetic pathway of erythromycin A in Streptomyces erythraeus. We report the crystal structures of EryK in complex with two widely used azole inhibitors: ketoconazole and clotrimazole. Both of these ligands use their imidazole moiety to coordinate the heme iron of P450s. Nevertheless, because of the different chemical and structural properties of their N1-substituent group, ketoconazole and clotrimazole trap EryK, respectively, in a closed and in an open conformation that resemble the two structures previously described for the ligand-free EryK. Indeed, ligands induce a distortion of the internal helix I that affects the accessibility of the binding pocket by regulating the kink of the external helix G via a network of interactions that involves helix F. The data presented thus constitute an example of how a cytochrome P450 may be selectively trapped in different conformational states by inhibitors.

Preliminary Experience with Abdominal Dual-energy CT (DECT): True Versus Virtual Nonenhanced Images of the Liver

La Radiologia Medica. Dec, 2010  |  Pubmed ID: 20852956

The aim of this work was to compare the quality and noise of true non-enhanced (TNE) and virtual non-enhanced (VNE) images in patients undergoing dual-energy computed tomography (DECT) of the liver.

Polarized X-ray Absorption Near-edge Structure Spectroscopy of Neuroglobin and Myoglobin Single Crystals

The Journal of Physical Chemistry. B. Oct, 2010  |  Pubmed ID: 20879741

Polarized Fe K-edge X-ray absorption near-edge structure (XANES) spectra of murine carbonmonoxy-neuroglobin (NgbCO) single crystals have been collected and compared with a number of derivatives of sperm whale myoglobin (Mb), that is, the nitrosyl (MbNO) and deoxy (Mb) derivatives, the previously reported cyanomet (MbCN) and carbonmonoxy (MbCO) derivatives, and the cryogenic photoproduct of MbCO (Mb·CO). The single crystals under study exhibit a strong XANES angular dichroism which allows the heme geometry of each sample to be analyzed with extremely high accuracy via the full multiple scattering (MS) approach. The results of two alternative methods to undergo the MS analysis have been compared with high resolution X-ray diffraction (XRD) data and with X-ray absorption spectroscopy (XAS) data in solution. As a result of the present analysis, the Fe-heme structure in solution and in the cryo-trapped NgbCO single crystal (which cracks at room temperature) are the same. Accordingly, the residual energy involved in the protein relaxation responsible of crystal cracking at room temperature after CO binding does not reside in the heme pocket. A combined approach (polarized XANES and XRD) is suggested to be applied on the same single crystals of metalloproteins at opportunely equipped synchrotron beamlines.

Evaluating Self-declared Ancestry of U.S. Americans with Autosomal, Y-chromosomal and Mitochondrial DNA

Human Mutation. Dec, 2010  |  Pubmed ID: 20886636

The current U.S. population represents an amalgam of individuals originating mainly from four continental regions (Africa, Europe, Asia and America). To study the genetic ancestry and compare with self-declared ancestry we have analyzed paternally, maternally and bi-parentally inherited DNA markers sensitive for indicating continental genetic ancestry in all four major U.S. American groups. We found that self-declared U.S. Hispanics and U.S. African Americans tend to show variable degrees of continental genetic admixture among the three genetic systems, with evidence for a marked sex-biased admixture history. Moreover, for these two groups we observed significant regional variation across the country in genetic admixture. In contrast, self-declared U.S. European and U.S. Asian Americans were genetically more homogeneous at the continental ancestry level. Two autosomal ancestry-sensitive markers located in skin pigmentation candidate genes showed significant differences in self-declared U.S. African Americans or U.S. European Americans, relative to their assumed parental populations from Africa or Europe. This provides genetic support for the importance of skin color in the complex process of ancestry identification.

Regression of Dilated Perivascular Spaces of the Brain

Surgical and Radiologic Anatomy : SRA. Jul, 2010  |  Pubmed ID: 19957183

Perivascular, or Virchow-Robin, spaces of the brain represent interstitial fluid-filled spaces continuous with subpial spaces, and not invagination of cerebrospinal fluid-filled subarachnoid spaces. Regression of a dilated, or even giant, perivascular space occurs rarely. The purpose of this paper is to describe magnetic resonance imaging evidence of complete regression of dilated perivascular spaces (dPVSs).

Granulomatous Reaction After an Aesthetic Electroporation Procedure

European Journal of Dermatology : EJD. Jan-Feb, 2010  |  Pubmed ID: 19959420

Mesenchymal Stromal Cells, Colony-forming Unit Fibroblasts, from Bone Marrow of Untreated Advanced Breast and Lung Cancer Patients Suppress Fibroblast Colony Formation from Healthy Marrow

Stem Cells and Development. Mar, 2010  |  Pubmed ID: 19388812

We have shown that bone marrow (BM) from untreated advanced lung and breast cancer patients (LCP and BCP) have a reduced number of colony-forming unit fibroblasts (CFU-Fs) or mesenchymal stem cells (MSCs). Factors that regulate the proliferation and differentiation of CFU-F are produced by the patients' BM microenvironment. We have now examined whether conditioned media (CM) from patients' CFU-F-derived stromal cells also inhibits the colony-forming efficiency (CFE) of CFU-F in primary cultures from healthy volunteers (HV)-BM. Thus the number and proliferation potential of HV-CFU-F were also found to be decreased and similar to colony numbers and colony size of patients' CFU-F. Stromal cells from both of these types of colonies appeared relatively larger and lacked the characteristic spindle morphology typically seen in healthy stromal cells. We developed an arbitrary mesenchymal stromal cell maturational index by taking three measures consisting of stromal cell surface area, longitudinal and horizontal axis. All stromal indices derived from HV-CFU-F grown in patients' CM were similar to those from stromal elements derived from patients' CFU-F. These indices were markedly higher than stromal indices typical of HV-CFU-F cultured in healthy CM or standard medium [alpha-medium plus 20% heat-inactivated fetal bovine serum (FBS)]. Patients' CM had increased concentrations of the CFU-F inhibitor, GM-CSF, and low levels of bFGF and Dkk-1, strong promoters of self-renewal of MSCs, compared to the levels quantified in CM from HV-CFU-F. Moreover, the majority of patients' MSCs were unresponsive in standard medium and healthy CM to give CFU-F, indicating that the majority of mesenchymal stromal cells from patients' CFU-F are locked in maturational arrest. These results show that alterations of GM-CSF, bFGF, and Dkk-1 are associated with deficient cloning and maturation arrest of CFU-F. Defective autocrine and paracrine mechanisms may be involved in the BM microenvironments of LCP and BCP.

Racial/ethnic Differences in Perceived Smoking Prevalence: Evidence from a National Survey of Teens

International Journal of Environmental Research and Public Health. Dec, 2010  |  Pubmed ID: 21318000

Prior studies show that perceived smoking prevalence is a significant predictor of smoking initiation. In this study, we examine racial/ethnic differences in perceived smoking prevalence and racial/ethnic differences in exposure to contextual factors associated with perceived smoking prevalence. We used cross-sectional time series data from the Legacy Media Tracking Surveys (LMTS), a national sample of 35,000 12- to 17-year-olds in the United States. Perceived smoking prevalence was the primary outcome variable, measured using an LMTS question: "Out of every 10 people your age, how many do you think smoke?" Multivariable models were estimated to assess the association between perceived smoking prevalence; race/ethnicity; and exposure to social contextual factors. Findings indicate that African American, Hispanic, and American Indian youth exhibit the highest rates of perceived smoking prevalence, while white and Asian youth exhibit the lowest. Minority youth are also disproportionately exposed to social contextual factors that are correlated with high perceived smoking prevalence. These findings suggest that disproportionate exposure to social contextual factors may partially explain why minority youth exhibit such high levels of perceived smoking prevalence.

Polarization Entangled State Measurement on a Chip

Physical Review Letters. Nov, 2010  |  Pubmed ID: 21231214

The emerging strategy to overcome the limitations of bulk quantum optics consists of taking advantage of the robustness and compactness achievable by integrated waveguide technology. Here we report the realization of a directional coupler, fabricated by femtosecond laser waveguide writing, acting as an integrated beam splitter able to support polarization-encoded qubits. This maskless and single step technique allows us to realize circular transverse waveguide profiles which are able to support the propagation of gaussian modes with any polarization state. Using this device, we demonstrate quantum interference with polarization-entangled states and singlet state projection.

Hyperentangled Mixed Phased Dicke States: Optical Design and Detection

Physical Review Letters. Dec, 2010  |  Pubmed ID: 21231568

A recently introduced family of multipartite entangled states, the 4-qubit phased Dicke states, has been created by 2-photon hyperentanglement. Our experimental method allows high state fidelity and generation rate. By introducing quantum noise in the multipartite system in a controlled way, we have tested the robustness of these states. To this purpose the entanglement of the resulting multipartite entangled mixed states has been verified by using a new kind of structural witness.

Prohibiting Menthol in Tobacco Products: a Policy Whose Time Has Come

Addiction (Abingdon, England). Dec, 2010  |  Pubmed ID: 21059131

Menthol and Non-menthol Smoking: the Impact of Prices and Smoke-free Air Laws

Addiction (Abingdon, England). Dec, 2010  |  Pubmed ID: 21059142

To examine the relationship between menthol and non-menthol prices and smoke-free air laws and the choice between menthol and non-menthol cigarettes among current smokers.

Evidence for Truth®: the Young Adult Response to a Youth-focused Anti-smoking Media Campaign

American Journal of Preventive Medicine. Dec, 2010  |  Pubmed ID: 21084069

Previous studies have shown that exposure to truth® and similar countermarketing campaigns is associated with an increase in anti-smoking attitudes and beliefs in those aged 12-17 years and a decrease in youth smoking. However, it is unclear how such campaigns influence young adults aged 18-24 years.

Integrated Photonic Quantum Gates for Polarization Qubits

Nature Communications. 2011  |  Pubmed ID: 22127062

The ability to manipulate quantum states of light by integrated devices may open new perspectives both for fundamental tests of quantum mechanics and for novel technological applications. However, the technology for handling polarization-encoded qubits, the most commonly adopted approach, is still missing in quantum optical circuits. Here we demonstrate the first integrated photonic controlled-NOT (CNOT) gate for polarization-encoded qubits. This result has been enabled by the integration, based on femtosecond laser waveguide writing, of partially polarizing beam splitters on a glass chip. We characterize the logical truth table of the quantum gate demonstrating its high fidelity to the expected one. In addition, we show the ability of this gate to transform separable states into entangled ones and vice versa. Finally, the full accessibility of our device is exploited to carry out a complete characterization of the CNOT gate through a quantum process tomography.

Experimental Realization of Optimal Noise Estimation for a General Pauli Channel

Physical Review Letters. Dec, 2011  |  Pubmed ID: 22243073

We present the experimental realization of the optimal estimation protocol for a Pauli noisy channel. The method is based on the generation of 2-qubit Bell states and the introduction of quantum noise in a controlled way on one of the state subsystems. The efficiency of the optimal estimation, achieved by a Bell measurement, is shown to outperform quantum process tomography.

Linkage Disequilibrium Analysis of D12S391 and VWA in U.S. Population and Paternity Samples

Forensic Science International. Genetics. Nov, 2011  |  Pubmed ID: 21067983

Recently, the European Network of Forensic Science Institutes voted to adopt five additional STR loci (D12S391, D1S1656, D2S441, D10S1248, and D22S1045) to their existing European Standard Set of seven STRs (TH01, vWA, FGA, D8S1179, D18S51, D21S11, and D3S1358). The D12S391 and vWA loci are located 6.3megabases (Mb) apart on chromosome 12. Ideally for use in forensic analyses, genetic markers on the same chromosome should be more than 50Mb in physical distance in order to ensure full recombination and thus independent inheritance. The purpose of this study was to evaluate if the closely located D12S391 and vWA loci are independent and, consequently, if these loci can be included in the product rule calculation for forensic and kinship analyses. Departures from Hardy-Weinberg equilibrium and linkage disequilibrium between the D12S391 and vWA loci were tested using n=654 unrelated U.S. African American, Caucasian, and Hispanic samples, and n=764 father/son paternity samples. In the unrelated U.S. population samples, no significant departures from HWE were detected for D12S391 or vWA. No significant evidence of linkage disequilibrium was observed between the loci in the population samples. However, significant linkage disequilibrium was detected in U.S. African American, Caucasian, and Asian father/son samples with phased genotypes. No significant linkage disequilibrium was detected for U.S. Hispanic paternity samples. The use of phased father/son pairs allowed for robust detection of linkage disequilibrium between D12S391 and vWA. In unrelated population samples, linkage disequilibrium is present but more difficult to detect due to the large number of possible haplotype combinations and unknown allelic phase. For casework analyses that involve unrelated or related individuals, the single-locus genotype probabilities for D12S391 and vWA should not be multiplied to determine the match probability of an autosomal STR profile. Since the D12S391 and vWA loci are not independent, it is recommended that the observed combination of alleles at D12S391 and vWA should be treated as a non-independent diplotype for profile probability calculations. The observed haplotype frequencies for U.S. African American, Caucasian, Hispanic, and Asian populations are provided for match probability calculations.

A Blind Circadian Clock in Cavefish Reveals That Opsins Mediate Peripheral Clock Photoreception

PLoS Biology. Sep, 2011  |  Pubmed ID: 21909239

The circadian clock is synchronized with the day-night cycle primarily by light. Fish represent fascinating models for deciphering the light input pathway to the vertebrate clock since fish cell clocks are regulated by direct light exposure. Here we have performed a comparative, functional analysis of the circadian clock involving the zebrafish that is normally exposed to the day-night cycle and a cavefish species that has evolved in perpetual darkness. Our results reveal that the cavefish retains a food-entrainable clock that oscillates with an infradian period. Importantly, however, this clock is not regulated by light. This comparative study pinpoints the two extra-retinal photoreceptors Melanopsin (Opn4m2) and TMT-opsin as essential upstream elements of the peripheral clock light input pathway.

Assessing Alternative Measures of Wealth in Health Research

American Journal of Public Health. May, 2011  |  Pubmed ID: 21252050

We assessed whether it would be feasible to replace the standard measure of net worth with simpler measures of wealth in population-based studies examining associations between wealth and health.

Contrast-enhanced Magnetic Resonance Imaging of Fibrovascular Tissue Ingrowth Within Synthetic Hydroxyapatite Orbital Implants in Children

European Journal of Ophthalmology. Sep-Oct, 2011  |  Pubmed ID: 21279974

To assess the usefulness of magnetic resonance imaging (MRI) in assessing fibrovascularization progression into synthetic hydroxyapatite (HA) implants inserted in anophthalmic sockets of children submitted to enucleation.

Neuroglobin-prion Protein Interaction: What's the Function?

Journal of Peptide Science : an Official Publication of the European Peptide Society. May, 2011  |  Pubmed ID: 21294227

Neuroglobin and cellular prion protein (PrP(C) ) are expressed in the nervous system and co-localized in the retinal ganglion cell layer. Both proteins do not have an unambiguously assigned function, and it was recently reported that PrP(C) aggregates rapidly in the presence of neuroglobin, whereas it does not aggregate in the presence of myoglobin, another globin with different tissue specificity. Electrostatic complementarity between the unstructured PrP(C) N-terminus and neuroglobin has been proposed to mediate this specific interaction. To verifythis hypothesis experimentally, we have used a combined approach of automated docking and molecular dynamics (MD) studies carried out on short stretches of prion protein (PrP) N-terminus to identify the minimal electrostatically interacting aminoacidic sequences with neuroglobin. Subsequently, we have performed the synthesis of these peptides by solid phase methods, and we tested their interaction with neuroglobin by surface plasmon resonance (SPR). Preliminary results confirm unequivocally the specific interaction between synthetic PrP peptides and neuroglobin suggesting a crucial role of PrP(C) positively charged regions in thisprotein-protein association.

Depictions of Tobacco Use in 2007 Broadcast Television Programming Popular Among US Youth

Archives of Pediatrics & Adolescent Medicine. Feb, 2011  |  Pubmed ID: 21300655

To determine the quantity of tobacco use in network television programming popular among US youth and to examine variation in tobacco depictions by TV Parental Guidelines system rating and television network.

The Light Responsive Transcriptome of the Zebrafish: Function and Regulation

PloS One. 2011  |  Pubmed ID: 21390203

Most organisms possess circadian clocks that are able to anticipate the day/night cycle and are reset or "entrained" by the ambient light. In the zebrafish, many organs and even cultured cell lines are directly light responsive, allowing for direct entrainment of the clock by light. Here, we have characterized light induced gene transcription in the zebrafish at several organizational levels. Larvae, heart organ cultures and cell cultures were exposed to 1- or 3-hour light pulses, and changes in gene expression were compared with controls kept in the dark. We identified 117 light regulated genes, with the majority being induced and some repressed by light. Cluster analysis groups the genes into five major classes that show regulation at all levels of organization or in different subset combinations. The regulated genes cover a variety of functions, and the analysis of gene ontology categories reveals an enrichment of genes involved in circadian rhythms, stress response and DNA repair, consistent with the exposure to visible wavelengths of light priming cells for UV-induced damage repair. Promoter analysis of the induced genes shows an enrichment of various short sequence motifs, including E- and D-box enhancers that have previously been implicated in light regulation of the zebrafish period2 gene. Heterologous reporter constructs with sequences matching these motifs reveal light regulation of D-box elements in both cells and larvae. Morpholino-mediated knock-down studies of two homologues of the D-box binding factor Tef indicate that these are differentially involved in the cell autonomous light induction in a gene-specific manner. These findings suggest that the mechanisms involved in period2 regulation might represent a more general pathway leading to light induced gene expression.

Successful Use of Ursodeoxycholic Acid in Nodular Regenerative Hyperplasia of the Liver

The Annals of Pharmacotherapy. Apr, 2011  |  Pubmed ID: 21450925

To report, to our knowledge, the first case of a patient with nodular regenerative hyperplasia of the liver (NRHL) associated with portal hypertension in whom ursodeoxycholic acid (UDCA) therapy had a therapeutic effect on liver enzymes and was associated with nonprogression of portal hypertension.

It's Time to Swim! Zebrafish and the Circadian Clock

FEBS Letters. May, 2011  |  Pubmed ID: 21486566

The zebrafish represents a fascinating model for studying key aspects of the vertebrate circadian timing system. Easy access to early embryonic development has made this species ideal for investigating how the clock is first established during embryogenesis. In particular, the molecular basis for the functional development of the zebrafish pineal gland has received much attention. In addition to this dedicated clock and photoreceptor organ, and unlike the situation in mammals, the clocks in zebrafish peripheral tissues and even cell lines are entrainable by direct exposure to light thus providing unique insight into the function and evolution of the light input pathway. Finally, the small size, low maintenance costs and high fecundity of this fish together with the availability of genetic tools make this an attractive model for forward genetic analysis of the circadian clock. Here, we review the work that has established the zebrafish as a valuable clock model organism and highlight the key questions that will shape the future direction of research.

Step Length Appears to Be a Strong Discriminant Gait Parameter for Elderly Females Highly Concerned About Falls: a Cross-sectional Observational Study

Physiotherapy. Jun, 2011  |  Pubmed ID: 21497246

To determine if gait parameters and the Timed Up and Go test can discriminate between elderly females with high and low concern about falls. Knowledge of these parameters could help in the development of rehabilitation programmes focused on the prevention of falls, fear of falling and functional decline.

Nodular Fasciitis of the Chest: Case Report of a Rare Presentation

Musculoskeletal Surgery. Dec, 2011  |  Pubmed ID: 21503721

Nodular fasciitis is a benign reactive proliferation that usually involves the deep fascia. The rapidly growing, the abundant cellularity, and the mitotic activity noted on histology can simulate an aggressive entity, such as sarcoma. We report an uncommon case of nodular fasciitis involved the chest wall of a thirteen boy and its management, emphasizing the need to consider this lesion in the differential diagnosis of pediatric soft tissue masses.

Effects of Lactobacillus Rhamnosus Strain GG in Pediatric Obesity-related Liver Disease

Journal of Pediatric Gastroenterology and Nutrition. Jun, 2011  |  Pubmed ID: 21505361

Various lines of evidence suggest that malfunctioning of the gut-liver axis contributes to hepatic damage of rodents and humans with nonalcoholic fatty liver disease. We evaluated the effects of short-term probiotic treatment in children with obesity-related liver disease who were noncompliant with lifestyle interventions.

A National Mass Media Smoking Cessation Campaign: Effects by Race/ethnicity and Education

American Journal of Health Promotion : AJHP. May-Jun, 2011  |  Pubmed ID: 21510785

To assess the effectiveness of a large-scale, national smoking cessation media campaign, the EX campaign, across racial/ethnic and educational subgroups.

Quit Attempts and Quit Rates Among Menthol and Nonmenthol Smokers in the United States

American Journal of Public Health. Jul, 2011  |  Pubmed ID: 21566032

We compared quit attempts and quit rates among menthol and nonmenthol cigarette smokers in the United States.

Modeling the Future Effects of a Menthol Ban on Smoking Prevalence and Smoking-attributable Deaths in the United States

American Journal of Public Health. Jul, 2011  |  Pubmed ID: 21566034

We used a validated smoking simulation model and data from the 2003 Tobacco Use Supplement to the Current Population Survey to project the impact that a US menthol ban would have on smoking prevalence and smoking-attributable deaths. In a scenario in which 30% of menthol smokers quit and 30% of those who would have initiated as menthol smokers do not initiate, by 2050 the relative reduction in smoking prevalence would be 9.7% overall and 24.8% for Blacks; deaths averted would be 633,252 overall and 237,317 for Blacks.

US Attitudes About Banning Menthol in Cigarettes: Results from a Nationally Representative Survey

American Journal of Public Health. Jul, 2011  |  Pubmed ID: 21566038

Menthol is a cigarette flavoring that makes smoking more appealing to smokers. The US Food and Drug Administration (FDA) has regulatory authority to ban mentholated cigarettes to reduce youth uptake and encourage adult cessation. Survey findings indicate that more than half of all Americans (56.1%) and of Blacks alone (68.0% in one sample and 75.8% in another) support banning menthol. Endorsement of a ban-especially by Blacks, who have the highest rates of menthol cigarette use-would support FDA action to ban menthol to protect the public's health.

Massive Hydrops of the Gallbladder Mimicking a Choledochal Cyst

Journal of Pediatric Surgery. May, 2011  |  Pubmed ID: 21616273

Preemptive Liver-kidney Transplantation in Von Gierke Disease: a Case Report

Transplantation Proceedings. May, 2011  |  Pubmed ID: 21620087

Type 1a glycogen storage disease (GSD 1a), or von Gierke disease, is a rare, autosomal-recessive disease caused by a deficiency of glucose-6-phosphatase, which leads to glycogen accumulation in the liver, kidney, and intestinal mucosa. Clinical manifestations include hypoglycemia, growth retardation, hepatomegaly, lactic acidemia, hyperlipidemia, and hyperuricemia. Long-term complications include renal disease, gout, osteoporosis, pulmonary hypertension, short stature, and hepatocellular adenomas, which may undergo malignant transformation. Herein we have described the management and the clinical course of a GSD1a patient who underwent simultaneous preemptive liver- kidney transplantation (SPLKT), which solved the liver and renal disease. We confirmed the rapid normalization of glucose metabolism, and correction of hyperlipemia after liver transplantation. In our opinion uremic patients with GSD 1a with or without adenomas must be considered for SPLKT. To our knowledge this is the fifth case of SPLKT and the first preemptive one to be described in the literature.

Tobacco: an Equal-opportunity Killer?

Tobacco Control. Jul, 2011  |  Pubmed ID: 21705468

Repair of Monteggia Fractures Using an Arthrex Tightrope System and Ulnar Plating

Veterinary Surgery : VS. Aug, 2011  |  Pubmed ID: 21770974

To report the outcome after treatment of Monteggia fractures (MF) using a novel surgical technique.

The Path to Quit: How Awareness of a Large-scale Mass-media Smoking Cessation Campaign Promotes Quit Attempts

Nicotine & Tobacco Research : Official Journal of the Society for Research on Nicotine and Tobacco. Nov, 2011  |  Pubmed ID: 21852272

Although awareness of mass-media smoking cessation campaigns is hypothesized to affect quit behavior through changes in cessation-related attitudes, intentions, and motivation (cognitions), this has yet to be formally tested.

Dual-source CT Coronary Angiography: Prospective Versus Retrospective Acquisition Technique

La Radiologia Medica. Mar, 2011  |  Pubmed ID: 20981504

The aim of our work was to compare image quality and radiation dose in a group of patients who underwent cardiac dual-source computed tomography (DSCT) with prospective electrocardiographic (ECG) gating with those of a control group studied with retrospective gating.

Evaluation of EX: a National Mass Media Smoking Cessation Campaign

American Journal of Public Health. Feb, 2011  |  Pubmed ID: 21164094

We used longitudinal data to examine the relationship between confirmed awareness of a national, branded, mass media smoking cessation campaign and cessation outcomes.

Implanted CardioWest-t Total Artificial Heart: Three-dimensional Computed Tomography Reconstruction

European Journal of Cardio-thoracic Surgery : Official Journal of the European Association for Cardio-thoracic Surgery. Jul, 2011  |  Pubmed ID: 21167731

CT and MRI of Wernicke's Encephalopathy

La Radiologia Medica. Mar, 2011  |  Pubmed ID: 21225366

The purpose of this pictorial essay is to present the computed tomography (CT) and magnetic resonance imaging (MRI) findings of Wernicke's encephalopathy, a rare, severe, acute neurological syndrome due to thiamine (vitamin B1) deficiency, associated with high morbidity and mortality. The classical clinical triad, which includes ocular signs, altered consciousness and ataxia, can be found in only one-third of patients. Although chronic alcoholic patients are the most commonly affected, Wernicke's encephalopathy may complicate malnutrition conditions in nonalcoholic patients, in whom it is greatly underestimated. CT and above all MRI of the brain play a fundamental role in diagnosing the condition and ruling out other diseases. MRI is the most sensitive technique and is required in all patients with a clinical suspicion of Wernicke's encephalopathy. Medial thalami, mamillary bodies, tegmentum, periaqueductal region, and tectal plate are typical sites of abnormal MRI signal. The dorsal medulla, red nuclei, cranial nerve nuclei, cerebellum, corpus callosum, frontal and parietal cerebral cortex are less common sites of involvement although they are more frequently affected in nonalcoholic patients. Paramagnetic contrast material may help to identify lesions not otherwise visible.

Autosomal SNP Typing of Forensic Samples with the GenPlex™ HID System: Results of a Collaborative Study

Forensic Science International. Genetics. Nov, 2011  |  Pubmed ID: 20650697

The GenPlex™ HID System (Applied Biosystems - AB) offers typing of 48 of the 52 SNPforID SNPs and amelogenin. Previous studies have shown a high reproducibility of the GenPlex™ HID System using 250-500pg DNA of good quality. An international exercise was performed by 14 laboratories (9 in Europe and 5 in the US) in order to test the robustness and reliability of the GenPlex™ HID System on forensic samples. Three samples with partly degraded DNA and 10 samples with low amounts of DNA were analyzed in duplicates using various amounts of DNA. In order to compare the performance of the GenPlex™ HID System with the most commonly used STR kits, 500pg of partly degraded DNA from three samples was typed by the laboratories using one or more STR kits. The median SNP typing success rate was 92.3% with 500pg of partly degraded DNA. Three of the fourteen laboratories counted for more than two thirds of the locus dropouts. The median percentage of discrepant results was 0.2% with 500pg degraded DNA. An increasing percentage of locus dropouts and discrepant results were observed when lower amounts of DNA were used. Different success rates were observed for the various SNPs. The rs763869 SNP was the least successful. With the exception of the MiniFiler™ kit (AB), GenPlex™ HID performed better than five other tested STR kits. When partly degraded DNA was analyzed, GenPlex™ HID showed a very low mean mach probability, while all STR kits except MiniFiler™ had very limited discriminatory power.

Ultrasound Contrast Media in Paediatric Patients: is It an Off-label Use? Regulatory Requirements and Radiologist's Liability

La Radiologia Medica. Feb, 2012  |  Pubmed ID: 21892716

The use of ultrasound contrast media is yet to be validated in children. The rare reports in the scientific literature denote their "sporadic" and "experimental" use. Their most likely use is in the setting of off-label prescription of medications. As there are still no guidelines available in Italy for the off-label use of medications, we aim to define the duties, obligations and liability of ultrasound radiologists according to the laws in force.

Extramedullary Intracranial Localization of Multiple Myeloma and Treatment with Novel Agents: A Retrospective Survey of 50 Patients

Cancer. Mar, 2012  |  Pubmed ID: 21932386

BACKGROUND: Intracranial involvement in multiple myeloma is extremely rare. The effect of new drugs (eg, thalidomide, bortezomib, lenalidomide) with respect to old drugs (eg, alkylators, steroids) has not been reported. METHODS: We collected clinical and biological data of patients presenting with an osteo-dural or primary dural multiple myeloma (OD-DMM) or a central nervous system myelomatosis (CNS-MM) by sending a questionnaire to the centers of the Gruppo Italiano Malattie Ematologiche dell'Adulto (GIMEMA). RESULTS: A total of 50 patients were registered. New therapies were used in 35 patients, whereas 15 patients received old treatments. Twenty-five out of 50 patients obtained a complete remission or a very good partial remission (CR+VGPR). Overall survival (OS) for CNS-MM was 6 months, for OD-DMM 25 months. OS was 25 months for patients treated with new agents versus 8 months with old agents. Improved OS and progression-free survival were predicted by response (CR+VGPR) and by patients who underwent stem cell transplantation versus chemotherapy. β2-Microglobulin >5 mmol/L was a poor prognostic factor. Multivariate analysis showed poor survival for patients with β2-microglobulin >5 mmol/L and better survival for patients achieving CR+VGPR. CONCLUSIONS: The overall data highlight the relevance of therapy with new drugs in intracranial myeloma, providing a framework for future clinical trials. Cancer 2011;. © 2011 American Cancer Society.

Primary and Dual Users of Cigars and Cigarettes: Profiles, Tobacco Use Patterns and Relevance to Policy

Nicotine & Tobacco Research : Official Journal of the Society for Research on Nicotine and Tobacco. Jan, 2012  |  Pubmed ID: 22259149

INTRODUCTION: The Family Smoking Prevention and Tobacco Control Act provides an unprecedented opportunity to regulate tobacco in the United States. However, restrictions on little cigars, cigarillos, and large cigars are notably absent from the Act, which may create a favorable environment for increased dual usage of cigars. The purpose of this study is to examine demographic profiles and tobacco use behaviors of dual, as opposed to primary, users of cigarettes and cigars. METHODS: This study used data from a cross-sectional nationally representative survey of 2,649 adult smokers and nonsmokers. Multivariable regression analysis was used to compare the demographic characteristics and tobacco use behaviors of dual users versus cigarette-only smokers. RESULTS: Data indicate that 12.5% of cigarette smokers are dual users of cigars. Dual users are more likely to be male, 18-29 years of age, non-Hispanic Black, of lower educational attainment, and either unemployed or out of the work force. Dual users were less likely than cigarette-only smokers to be daily cigarette smokers (odds ratio [OR] = 0.57, 95% CI: 0.32, 1.02), more likely to have made a recent quit attempt (OR = 2.39, 95% CI: 1.44, 3.97), and more likely to have used at least one other alternative product (OR = 2.26, 95% CI: 1.26, 4.05), including snus, e-cigarettes, dissolvables, and chewing tobacco.Conclusions:As greater restrictions on cigarettes become implemented in the United States, it will be critical to monitor increased dual use of cigars, in order to inform prevention and treatment strategies and guide more comprehensive policy efforts.

Superselective Ophthalmic Artery Infusion of Melphalan for Intraocular Retinoblastoma: Preliminary Results from 140 Treatments

Acta Ophthalmologica. Jan, 2012  |  Pubmed ID: 22268993

Purpose:  To report our experience in superselective ophthalmic artery infusion of melphalan (SOAIM) for intraocular retinoblastoma. Methods:  From June 2008 to October 2010, 38 patients (18 women, 20 men; age range at first treatment, 7 months to 22 years) with 41 eyes with retinoblastoma were scheduled for SOAIM, for 17 newly diagnosed retinoblastomas Tumour, Node and Metastasis (TNM) 7th Edition 1a (n = 1), 1b (n = 1), 2a (n = 7), 2b (n = 4) and 3a (n = 4) and 24 retinoblastomas with partial remission/relapse TNM 7th Edition 1b (n = 13), 2a (n = 1) and 2b (n = 10). Eight patients (ten eyes) have been treated by SOAIM alone. Follow-up was 6-27 months in 28 patients (30 eyes). Results:  Ophthalmic artery cannulation failed in two patients. Thirty-six patients underwent 140 treatments by internal (n = 112) or external (n = 28) carotid arteries. No major procedural complications occurred. Two patients have been lost to follow-up. Remaining 34 patients (37 eyes) had no metastatic disease. Four patients suffered permanent ocular complications: chorioretinal dystrophy (n = 2), ptosis (n = 1) and strabismus/exotropia (n = 1). Eight (22%) eyes in eight (24%) patients underwent enucleation: 7/16 (43%) newly diagnosed retinoblastomas and 1/22 (4.5%) retinoblastomas undergoing partial remission/relapse. For all treated eyes, Kaplan-Meier eye enucleation-free rates (K-M) were 85.4% (95% CI, 73.3-97.5%), 74.4% (95% CI, 57-91.8%) and still stable at 6, 12 months and 2 years, respectively. For eyes with partial remission/relapse, and eyes at presentation, K-M at 2 years were 95.5% (95% CI, 86.9-100%) and 45.6% (95% CI, 16.6-74.6%), respectively. Conclusion:  Superselective ophthalmic artery infusion of melphalan was safe and powerful, especially following other therapies. Superselective ophthalmic artery infusion of melphalan should be added to focal therapies spectrum. In selected cases, melphalan should be combined with other chemotherapeutic agents.

Two-particle Bosonic-fermionic Quantum Walk Via Integrated Photonics

Physical Review Letters. Jan, 2012  |  Pubmed ID: 22304249

Quantum walk represents one of the most promising resources for the simulation of physical quantum systems, and has also emerged as an alternative to the standard circuit model for quantum computing. Here we investigate how the particle statistics, either bosonic or fermionic, influences a two-particle discrete quantum walk. Such an experiment has been realized by exploiting polarization entanglement to simulate the bunching-antibunching feature of noninteracting bosons and fermions. To this scope a novel three-dimensional geometry for the waveguide circuit is introduced, which allows accurate polarization independent behavior, maintaining remarkable control on both phase and balancement.

Global Tobacco Inequalities: a New Frontier

Cancer Causes & Control : CCC. Mar, 2012  |  Pubmed ID: 22367723

Insulin Sensitivity Deteriorates After Short-term Lifestyle Intervention in the Insulin Sensitive Phenotype of Obesity

Obesity Facts. 2012  |  Pubmed ID: 22433619

Objective: To investigate the effects of a 3-month lifestyle intervention on insulin sensitivity and its related cardiometabolic factors in obese patients. Methods: Anthropometry, body composition, oral glucose tolerance test, lipids, alanine aminotransferase, insulin sensitivity (insulinogenic index (ISI), homeostasis model assessment, β-cell performance (disposition index)) were evaluated in 263 obese women and 93 obese men before and after 3 months of hypocaloric low fat/high protein diet associated with physical activity 30 min/day. Results: Patients were divided into 3 groups according to the intervention-induced ISI changes: group 1 (decrease), group 2 (stability) and group 3 (increase). Insulin sensitivity and the disposition index were significantly higher before the intervention in group 1 than in group 3. BMI, waist circumference, and fat mass significantly decreased in groups 1 and 3 in both sexes. β-cell performance decreased in group 1 and increased in group 3. Metabolic variables improved in group 3, whereas glucose levels increased in women of group 1. The post-intervention insulin sensitivity was lower in group 1 than in group 3. Conclusion: Lifestyle intervention induces changes in insulin sensitivity and metabolic factors that depend on the pre-intervention degree of insulin sensitivity. Weight loss leads to metabolic benefits in insulin-resistant, obese patients, whereas it may paradoxically worsen the metabolic conditions in the insulin-sensitive phenotype of obesity. Copyright © 2012 S. Karger GmbH, Freiburg.

Capillary Electrophoresis of an 11-plex MtDNA Coding Region SNP Single Base Extension Assay for Discrimination of the Most Common Caucasian HV1/HV2 Mitotype

Methods in Molecular Biology (Clifton, N.J.). 2012  |  Pubmed ID: 22139659

The typing of single nucleotide polymorphisms (SNPs) located throughout the human mitochondrial genome assists in resolving individuals with an identical HV1/HV2 haplotype. A set of 11 sites which were selected for distinguishing individuals of a common Western European Caucasian HV1/HV2 mitotype was incorporated into a single base extension (SBE) assay. The assay was optimized for multiplex detection of sequence polymorphisms at positions 3010, 4793, 10211, 5004, 7028, 7202, 16519, 12858, 4580, 477, and 14470 in the mitochondrial genome. PCR primers were designed to allow for multiplex amplification of unique regions in the mitochondrial genome followed by an 11-plex SBE reaction using the SNaPshot(®) reagent kit. Separation and detection can be accomplished with a capillary-based electrophoresis platform commonly found in most forensic laboratories.