Alpha-helix to beta-sheet transition in long-chain poly-l-lysine: Formation of alpha-helical fibrils by poly-l-lysine.
The temperature-induced α-helix to β-sheet transition in long-chain poly-l-lysine (PLL), accompanied by the gauche-to-trans isomerization of CH2 groups in the hydrocarbon side chains of Lys amino acid residues, and formation of β-sheet as well as α-helix fibrillar aggregates of PLL have been studied using Fourier-transform infrared (FT-IR) and vibrational circular dichroism (VCD) spectroscopy, and transmission electron microscopy (TEM). In a low-temperature alkaline water solution or in a methanol-rich water mixture, the secondary structure of PLL is represented by α-helical conformations with unordered and gauche-rich hydrocarbon side chains. Under these conditions, PLL molecules aggregate into α-helical fibrils. PLLs dominated by extended antiparallel β-sheet structures with highly ordered trans-rich hydrocarbon side chains are formed in a high-temperature range at alkaline pD and aggregate into fibrillar, protofibrillar, and spherical forms. Presented data support the idea that fibrillar aggregation is a varied phenomenon possible in repetitive structural elements with not only a β-sheet-rich conformation, but also an α-helical-rich conformation.


Weight loss after laparoscopic adjustable gastric banding is not caused by altered gastric emptying.
In order to know the role of gastric emptying in the mechanism of weight loss and early satiety after a restrictive surgical procedure for treatment of morbid obesity, a consecutive series of patients were scintigraphically investigated before and after laparoscopic adjustable gastric banding. Sixteen patients undergoing laparoscopic adjustable gastric banding underwent preoperatively, and at 6 months postoperatively, a gastric emptying study (solid meal and single isotope). Esophageal retention time, lag phase, peak activity time, gastric emptying rate, fundus emptying rate, and weight loss were recorded. Upper GI symptom assessment was carried out by using a standardized questionnaire. Gastric emptying parameters were correlated with the upper GI symptoms. Gastric band placement showed no significant influence on postoperative gastric emptying rate [median % (interquartile range): 42 (23.3-59) preoperatively vs 38 (31-71) postoperatively and fundus emptying rate: 59(37-91) preoperatively vs 70 (53-89) postoperatively]; however, an increase in early satiety was found. Neither gastric emptying rate nor fundus emptying rate showed a relation with early satiety or weight loss. Furthermore, no correlation was found between early satiety and lag phase, esophageal retention time, start of activity, and peak activity time in proximal stomach. Laparoscopic adjustable gastric banding seems not to affect gastric emptying. Neither a relation between postoperative gastric emptying rate and weight loss nor between early satiety and weight loss was found. Therefore, it is unlikely that gastric emptying plays a role in the mechanism of weight loss following laparoscopic adjustable gastric banding.


Is There a Shift in Salmonella Diversity Among Poultry in Northern India?
The present study was conducted to determine the serotype diversity of Salmonella among poultry in northern India. A total of 101 poultry droppings from 30 farms in the Jammu and Kashmir union territory were analyzed. Nineteen isolates of Salmonella were obtained, and these belonged to four serotypes: Salmonella enterica enterica serotype Kentucky (n = 3), Salmonella enterica enterica serotype Infantis (n = 5), Salmonella enterica enterica serotype Agona (n = 4), and Salmonella enterica enterica serotype Typhimurium (n= 7). The study has isolated some Salmonella serotypes that are infrequently reported in India. Some of the isolated serotypes are reported to be endemic for human nontyphoidal salmonellosis cases in the region. Whether this indicates a shift in the serotype pattern in poultry in the region needs to be investigated further. Nevertheless, the study clearly indicates the risk of foodborne salmonellosis associated with consumption of contaminated poultry and poultry products in the region.


Genetic iron overload disorders.
Due to its pivotal role in orchestrating vital cellular functions and metabolic processes, iron is an essential component of the human body and a main micronutrient in the human diet. However, excess iron causes an increased production of reactive oxygen species leading to cell dysfunction or death, tissue damage and organ disease. Iron overload disorders encompass a wide spectrum of pathological conditions of hereditary or acquired origin. A number of 'iron genes' have been identified as being associated with hereditary iron overload syndromes, the most common of which is hemochromatosis. Although linked to at least five different genes, hemochromatosis is recognized as a unique syndromic entity based on a common pathogenetic mechanism leading to excessive entry of unneeded iron into the bloodstream. In this review, we focus on the pathophysiologic basis and clinical aspects of the most common genetic iron overload syndromes in humans.


Dapsone Therapy for Pustular Psoriasis: Case Series and Review of the Literature.
Pustular psoriasis is an uncommon psoriasis variant, clinically characterized as small sterile pustules on an erythematous base. Evidence for therapy is lacking, and many currently employed systemic therapeutics carry risks of significant side effects, without specifically targeting disease etiology which includes the aggregation of neutrophils. We report therapy with the anti-neutrophil agent dapsone in 5 patients with pustular psoriasis and provide a brief review of the literature. Four patients responded to oral dapsone and 1 to topical dapsone therapy. All 5 patients had previously failed multiple topical and systemic treatments. In 2 cases, oral dapsone allowed for the discontinuation of other systemic agents. One patient stopped oral dapsone due to a side effect of sleep disturbance. Dapsone has a much safer side effect profile and may target the pathophysiology of pustular psoriasis more directly than many other systemic agents. As such, dapsone should be considered for the treatment of patients with pustular psoriasis.


Chlamydial infection: an accurate model for opportunistic screening in general practice.
To estimate the prevalence of Chlamydia trachomatis in women in general practice and to assess risk factors associated with infection. The study was carried out in 2001-2 in different general practices in Antwerp, Belgium. Sexually active women, visiting their general practitioner for routine gynaecological care (mostly pill prescription or PAP smear), were offered opportunistic screening for chlamydia. 787 participants aged 15-40 delivered a self taken vaginal sample and filled in a questionnaire which included questions on demographic variables, urogenital symptoms, sexual history, and sexual behaviour. Samples were tested for presence of chlamydial DNA by means of a ligase chain reaction (LCR) assay, and positives were confirmed by two other amplification assays (PCR and SDA). Overall prevalence was 5.0% (95% CI: 3.5 to 6.5). Determinants of infection in logistic regression analysis were age 18-27 years, >1 partner in the past year, no use of contraceptives, frequent postcoital bleeding, having a symptomatic partner, painful micturition, and living in the inner city. The area under the ROC curve in the full model was 0.88. Selective screening based on a combination of the five first determinants detects 92.3% of infections in this sample; 37.5% of the population would need to be screened. Targeted screening for chlamydial infection is possible, even in a heterogeneous group of general practice attendants. Implementing this model would require considerable communication skills from healthcare providers.


Studies on the oxidation reaction of tyrosine (Tyr) with H2O2 catalyzed by horseradish peroxidase (HRP) in alcohol-water medium by spectrofluorimetry and differential spectrophotometry.
An oxidation reaction of tyrosine (Tyr) with H(2)O(2) catalyzed by horseradish peroxidase (HRP) was studied by spectrofluorimetry and differential spectrophotometry in the alcohol(methanol, ethanol, 1-propanol and isopropanol)-water mutual solubility system. Compared with the enzymatic-catalyzed reaction in the water medium, the fluorescence intensities of the product weakened, even extinguished. Because the addition of alcohols made the conformation of HRP change, the catalytic reaction shifted to the side of polymerization and the polymer (A(n)H(2), n>or=3) exhibited no fluorescence. The four alcohols cannot deactivate HRP. Moreover isopropanol activated HRP remarkably.


Evaluation of hypocrellin A-loaded lipase sensitive polymer micelles for intervening methicillin-resistant Staphylococcus Aureus antibiotic-resistant bacterial infection.
There is an urgent need for new antibacterial strategies to overcome the emergence of antibiotic resistance. Antibacterial photodynamic therapy (APDT) may be an effective method to deliver photosensitizers for the treatment of methicillin-resistant Staphylococcus aureus (MRSA) infections. Here, we report that the photosensitizer hypocrellin A (HA) loaded into lipase-sensitive methoxy poly (ethylene glycol)-block-poly(ε-caprolactone) (mPEG-PCL) micelles showed high anti-MRSA activity in vitro and in vivo by PDT. Once the micelles come into contact with bacteria that secrete lipase, the PCL is degraded to release HA. Our results showed that the minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of mPEG-PCL/HA micelles after light irradiation were 0.69 and 1.38 mg/L (HA concentration), respectively. In the dark, the MIC and MBC of the micelles were 250 and 500 mg/L (HA concentration), respectively. The fluorescent stain results further demonstrated the photodynamic antibacterial activity of mPEG-PCL/HA micelles. The survival rate of mice subjected to experimental acute peritonitis increased to 86% after treated with the micelles. The polymeric micelles showed low hemolytic activity and biocompatibility, simultaneously preventing aggregation in vivo and enhancing the water solubility of HA. Thus, the photosensitizer HA loaded micelles could be used as APDT for infections caused by bacteria without antibiotic resistance.


Perioperative circulating tumor cells in surgical patients with non-small cell lung cancer: does surgical manipulation dislodge cancer cells thus allowing them to pass into the peripheral blood?
We herein evaluated the status of circulating tumor cells (CTC) dislodged from the tumor during surgery in patients who underwent pulmonary resection for non-small cell lung cancer (NSCLC) to assess the clinical implications. Tumor cells in the peripheral arterial blood before surgery (Before) and immediately after lung resection (After) and in the blood from the pulmonary vein of the resected lung were detected using a size selective method. The clinicopathological characteristics and the prognosis were then analyzed according to the CTC status: no tumor cells detected (N), single tumor cell or total number less than 4 cells (S), and existence of clustered cells (C). According to the CTC status, the patients were classified into the following three groups: Before-C and After-C, Group I (n = 6); Before-S or N and After-C, Group II (n = 9); and Before-S or N and After-S or N, Group III (n = 8). Group III showed a high rate of p-stage IA, smaller tumor size, lower CEA level, lower SUVmax level, and a higher relapse-free survival rate than the other groups. CTCs were detected in patients after undergoing lung resection, some of which may have been dislodged by the surgical procedure. The presence of clustered CTCs after the operation indicated an unfavorable outcome.


Performance of comorbidity, risk adjustment, and functional status measures in expenditure prediction for patients with diabetes.
To compare the ability of generic comorbidity and risk adjustment measures, a diabetes-specific measure, and a self-reported functional status measure to explain variation in health care expenditures for individuals with diabetes. This study included a retrospective cohort of 3,092 diabetic veterans participating in a multisite trial. Two comorbidity measures, four risk adjusters, a functional status measure, a diabetes complication count, and baseline expenditures were constructed from administrative and survey data. Outpatient, inpatient, and total expenditure models were estimated using ordinary least squares regression. Adjusted R(2) statistics and predictive ratios were compared across measures to assess overall explanatory power and explanatory power of low- and high-cost subgroups. Administrative data-based risk adjusters performed better than the comorbidity, functional status, and diabetes-specific measures in all expenditure models. The diagnostic cost groups (DCGs) measure had the greatest predictive power overall and for the low- and high-cost subgroups, while the diabetes-specific measure had the lowest predictive power. A model with DCGs and the diabetes-specific measure modestly improved predictive power. Existing generic measures can be useful for diabetes-specific research and policy applications, but more predictive diabetes-specific measures are needed.


Hepatic microRNAome reveals potential microRNA-mRNA pairs association with lipid metabolism in pigs
As one of the most important metabolic organs, the liver plays vital roles in modulating the lipid metabolism. This study was to compare miRNA expression profiles of the Large White liver between two different developmental periods and to identify candidate miRNAs for lipid metabolism. Eight liver samples were collected from White Large of 70-day fetus (P70) and of 70-day piglets (D70) (with 4 biologocal repeats at each development period) to construct sRNA libraries. Then the eight prepared sRNA libraries were sequenced using Illumina next-generation sequencing techonogy on HiSeq 2500 platform. As a result, we obtained 346 known and 187 novel miRNAs. Compared with the D70, 55 down- and 61 up-regulated miRNAs were shown to be significantly differentially expressed (DE). GO and KEGG enrichment analysis indicated that these DE miRNAs were mainly involved in growth, development and diverse metabolic processes. They were predicted to regulate lipid metabolism through Adipocytokine signaling pathway, MAPK, AMPK, cAMP, PI3K-AKT, and Notch signaling pathway. miR-122, miR-26a and miR-30a-5p, which play important roles in lipid metabolism, were the most abundantly expressed (miR-122 only in P70). Integration analysis (details of mRNAs sequencing data were shown in another unpublished paper) revealed that many target genes of the DE miRNAs (miR-181b, miR-145-5p, miR-199a-5p and miR-98) might be critical regulators in lipid metabolic process, including ACSL4, ABCA4 and SCD. Thus, these miRNAs were considered to be the promising candidates for lipid metabolism. Our study provides the main differences in the Large White at miRNA level between two different developmental stages. It supplies a valuable database for the further function and mechanism elucidation of miRNAs in porcine liver development and lipid metabolism.


Phospholipid exchange between subcellular organelles of rabbit lung.
A soluble protein fraction (PLEP) prepared from rabbit lung can catalyze the exchange of phospholipids between subcellular organelles of the lung and between these subcellular organelles and synthetic liposomes. Phospholipid exchange between microsomes and synthetic liposomes and between mitochondria and synthetic liposomes was stimulated 8-fold and 2.5-fold, respectively, in the presence of the protein fraction. Lung exchange protein could also catalyzed phospholipid exchange between subcellular organelles of the liver and synthetic liposomes. Phospholipid transfer between microsomes and lamellar bodies of the lung was stimulated 2-fold by the exchange protein. Both radiolabeled phosphatidylcholine (PC) and phosphatidylinositol (PI) were transferred from 32P-labeled microsomes to lamellar bodies, but the exchange protein exhibited no transfer activity for phosphatidylglycerol (PG) and that for phosphatidylethanolamine (PE) was insignificant compared to the transfer activity for phosphatidylcholine and phosphatidylinositol. While the physiological role of the phospholipid exchange proteins in the lung is unknown, it is possible that they participate in the distribution of the newly synthesized phospholipids from the site of synthesis to lamellar bodies and other membrane compartments of cells.


Effectiveness of triennial screening with clinical breast examination: 14-years follow-up outcomes of randomized clinical trial in Trivandrum, India.
This study presents the preliminary results of a randomized controlled trial (RCT) initiated in January 2006 in India to evaluate the effectiveness of clinical breast examination (CBE) in reducing breast cancer mortality as compared to a no-screening control group reported significant downstaging in the intervention group. The present manuscript reports long-term follow-up outcomes. Women 30-69 years old from 133 intervention clusters and 141 control clusters were invited to participate. Women in the intervention arm underwent three rounds of CBE every 3 years. CBE-positive women were reexamined by a physician, and triple-assessment was performed on those confirmed to have abnormalities. All participants were followed through home visits and linkage with population-based cancer registry. Of the 55,843 eligible women in the intervention arm, 95.7% had CBE at least once and 11.5% were CBE-positive. Breast cancers were diagnosed in 335 participants in the intervention group and 273 in the control group (N = 59,447). Age-standardized incidence rate of early cancer was 30.4 of 100,000 in the intervention and 21.9 of 100,000 in the control group, with a rate ratio (RR) of 1.4 (95% confidence interval [CI], 1.1-1.8). The age-standardized breast cancer mortality rates were 11.3 and 11.1 per 100,000 in intervention and control arms, respectively (RR, 1.1; 95% CI, 0.8-1.5) after 15 years. Five-year breast cancer survival rates were 77.0% in the intervention and 71.2% in the control groups (overall p value = .043). Triennial CBE screening failed to demonstrate any mortality benefit despite achieving a shift toward earlier stage at detection and improved survival in the intervention arm. CBE is a valuable tool for diagnosis of breast cancer in symptomatic women especially in areas where mammography and/or breast cancer screening programs are not widely available.


Perforation of Meckel's Diverticulum by a Peanut Presenting as a Mesentery Abscess.
Perforation of Meckel's diverticulum by a foreign body in children is rarely reported and is usually associated with localized or generalized fibropurulent peritonitis. The authors encountered such a case in a 4-year-old boy with perforation of Meckel's diverticulum by a piece of peanut presenting as a mesentery abscess. The diverticulum was wide-based and histology showed a transmural perforation at the tip of the diverticulum. Local inflammation due to irritation of the foreign body and progressive pressure necrosis at the tip of the diverticulum may be the pathogenesis of mesentery abscess. A search of the English literature did not reveal any similar case.


Expression of microRNAs in the urine of patients with bladder cancer.
We quantified the urine sediment and supernatant levels of microRNA (miRNA) targets related to epithelial-mesenchymal transition in 51 patients with bladder cancer and in 24 controls. We found that patients with bladder cancer had depressed levels of the miR-200 family, miR-192, and miR-155 in urinary sediment. The urinary level of these miRNAs may be developed as noninvasive markers for bladder cancer. MicroRNAs (miRNA) have been implicated to play an important role in the pathogenesis of a variety of cancers. We studied the levels of miRNAs related to epithelial-mesenchymal transition (EMT) in the urine of patients with bladder cancer. The expression of the miR-200 family, miR-205, miR-192, miR-155, and miR-146a in the urine sediment and supernatant of 51 patients with bladder cancer and in 24 controls was determined by real-time quantitative polymerase chain reaction. Compared with controls, the patients with bladder cancer had a lower expression of the miR-200 family, miR-192, and miR-155 in the urinary sediment; lower expression of miR-192; and higher expression of miR-155 in the urinary supernatant. The expression of the miR-200 family, miR-205, and miR-192 in the urine sediment significantly correlated with urinary expression of EMT markers, including zinc finger E-box-binding homeobox 1, vimentin, transforming growth factor β1, and Ras homolog gene family, member A. Furthermore, the levels of miR-200c and miR-141 in the urine sediment became normalized after surgery. We found that the urinary miR-200 family, miR-155, miR-192, and miR-205 levels are depressed in patients with bladder cancer. The level of these miRNA targets in urine has the potential to be developed as noninvasive markers for bladder cancer.


Becker muscular dystrophy: case report, review of the literature, and analysis of differentially expressed hub genes.
Becker muscular dystrophy (BMD) is a genetic and progressive neuromuscular disease caused by mutations in the dystrophin gene with no available cure. A case report and comprehensive review of BMD cases aim to provide important clues for early diagnosis and implications for clinical practice. Genes and pathways identified from microarray data of muscle samples from patients with BMD help uncover the potential mechanism and provide novel therapeutic targets for dystrophin-deficient muscular dystrophies. We describe a BMD family with a 10-year-old boy as the proband and reviewed BMD cases from PubMed. Datasets from the Gene Expression Omnibus database were downloaded and integrated with the online software. The systematic review revealed the clinical manifestations and mutation points of the dystrophin gene. Gene ontology analysis showed that extracellular matrix organization and extracellular structure organization with enrichment of upregulated genes coexist in three datasets. We present the first report of TUBA1A involvement in the development of BMD/Duchenne muscular dystrophy (DMD). This study provides important implications for clinical practice, uncovering the potential mechanism of the progress of BMD/DMD, and provided new therapeutic targets.


Comparison of microbiological risk assessment studies published.
The FAO/WHO risk analysis framework and principles are in the process of being implemented in different national and international settings. The importance of a further development of international co-operation based upon agreed principles in this area is stressed in the Codex Alimentarius system, as well as under the WTO/SPS agreements. Key input in this context will come from the presentation of formalised microbiological risk assessments. The number of peer-review published microbiological risk assessments is low and the format is still variable. This paper presents a comparison of selected risk assessments in the microbiological area, and suggests a number of lessons to be learned from these.


Phenotypic consequences of branch point substitutions.
The branch point sequence (BPS) is a conserved splicing signal important for spliceosome assembly and lariat intron formation. BPS mutations may result in aberrant pre-mRNA splicing and genetic disorders, but their phenotypic consequences have been difficult to predict, largely due to a highly degenerate nature of the BPS consensus. Here, we have examined the splicing pattern of nine reporter pre-mRNAs that have previously been shown to give rise to human hereditary diseases as a result of single-nucleotide substitutions in the predicted BPS. Increased exon skipping and intron retention observed in vivo were recapitulated for each mutated pre-mRNA, but the reproducibility of cryptic splice site activation was lower. BP mutations in reporter pre-mRNAs frequently induced aberrant 3' splice sites and also activated a cryptic 5' splice site. Systematic mutagenesis of BP adenosines showed that in most pre-mRNAs, the expression of canonical transcripts was lower for BP transitions than BP transversions. Differential splicing outcome for transitions vs. transversions was abrogated or reduced if introns were truncated to 200 nt or less, suggesting that the nature of the BP residue is less critical for interactions across very short introns. Together, these results improve prediction of phenotypic consequences of point mutations upstream of splice acceptor sites and suggest that the overrepresentation of disease-causing adenosine-to-guanosine BP substitutions observed in Mendelian disorders is due to more profound defects of gene expression at the level of pre-mRNA splicing.


Increased copper toxicity in Saccharomyces cerevisiae lacking VPS35, a component of the retromer and monogenic Parkinson disease gene in humans.
The Saccharomyces cerevisiae gene VPS35 encodes a component of the retromer complex which is involved in vesicle transport from endosomes to the trans-Golgi network. Yeast and human VPS35 orthologs are highly conserved and mutations in human VPS35 cause an autosomal dominant form of late-onset Parkinson disease (PD). We now show that deletion of VPS35 in yeast (vps35Δ) leads to a dose-dependent growth defect towards copper. This increased sensitivity could be rescued by transformation with yeast wild-type VPS35 but not by the expression of a construct harboring the yeast equivalent (i.e. D686N) of the most commonly identified VPS35-associated PD mutation, p.D620N. In addition, we show that expression of one copy of α-synuclein, which is known to directly interact with copper, leads to a pronounced aggravation of copper toxicity in vps35Δ cells, thereby linking the regulation of copper homeostasis by Vps35p in yeast to one of the key molecules in PD pathophysiology.


Anthropology and the theory of medicine.
Instead of presupposing reality as the "realists" do, a genetic theory of knowledge attempts to understand scientific knowledge through the psychological origins of both the concepts, and the operations on which these concepts are based. Adopting the viewpoint of genetic epistemology, the envisaged theory of medicine will have to perform a threefold task: (1) A revision and reformulation of the psycho-physical problem and the development of a model for the living body; (2) A revision of our views concerning the relationship between humans and the inanimate and animate environment; and (3) The development of a concept for the construction of individual reality and its consequences for the patient-physician relationship.


Intervening sequences in human fetal globin genes adopt left-handed Z helices.
The large intervening sequences ( IVS2 ) of three human fetal globin genes contain tracts of alternating purine-pyrimidine sequences approximately 40-60 base pairs in length which adopt left-handed Z DNA helices under the influence of negative supercoiling. The amount of negative supercoiling (approximately 0.045) required for the right- to left-handed transitions is within the physiological range. The structural aberrations between the right- and left-handed helices were mapped by sequencing the S1 nuclease cleavage sites. Two-dimensional gel electrophoretic analyses of the supercoil-induced relaxation served to characterize the type and length of left-handed structure. Furthermore, binding studies with several types of antibodies confirmed the presence of left-handed helices. Since these simple sequences appear to be hotspots for recombination and gene conversion, unusual DNA conformations may participate in genetic expression.


The effect of various S-alkylating agents on the chromatographic behavior of cysteine-containing peptides in reversed-phase chromatography.
We investigate the influence of various alkylation chemistries on the reversed phase (RP) HPLC behavior of Cys-containing peptides under the most popular RP-HPLC conditions used in proteomics: C18 phases with trifluoroacetic acid (TFA) or formic acid (FA) as the ion pairing modifiers, and separation at pH 10. Akylating agents studied are iodoacetamide (IAM), iodoacetic acid (IAA), 4-vinylpyridine (4-VP), acrylamide (AA) and methyl methanethiosulfonate (MMTS). These were compared against the retention of identical peptides without alkylation, i.e. free cysteines. The intrinsic hydrophobicity values of the Cys residue under formic acid conditions for these modifications were found to increase in the following order: 4-VP<IAM<AA<IAA<free Cys<MMTS. The retention contribution of the positively charged 4-VP derivative is affected by the nature of the ion-pairing modifier; this is the most hydrophilic residue for formic acid based eluents, and second most hydrophilic behind IAM-alkylated Cys using TFA eluent. Switching to a basic condition dramatically decreases the retention of free cysteine and IAA-alkylated analytes due to the ionization of side-chains. The opposite effect is observed for 4-VP, which become neutral at basic pHs. The careful measurement of the hydrophobic contributions for these residues is vital to the development of accurate peptide retention prediction models; the incorporation of these modifications into our Sequence Specific Retention Calculator model is presented.


A Study of Toluidine Blue Staining in Suspected Oral Malignancies in Patients Presenting to Tertiary Care Hospital in Central India.
Oral malignancies are a major cause of morbidity and mortality in worldwide and especially in Indian subcontinent. India stands at first position in total number of oral cancers in worldwide. Hence it is the need of hour to have safe and reliable screening test for early diagnosis of oral malignancies. Toluidine blue (TB) staining is an ideal screening test with high sensitivity and specificity. This study carried out in our department with objective to study efficacy of TB in oral malignancies and potential oral malignant lesions and also to evaluate and compare TB staining with histopathological examination of lesion. This prospective study conducted on 183 patients who attended ENT department. Out of 183 patients 146 (79.8%) were positive with TB and 37 (20.2%) were not stained with TB. Out of 146 positive patients 134 were turned out to be positive for malignancy in histopathological examination. Out of 37 negative patients 5 were positive for malignancy in histopathological examination and 32 were negative. Out of 183 patients 139 (76%) were positive whereas 44 (24%) were found negative for malignancy. In present study clinical examination of oral cavity reveal most of the patients had leukoplakis (37.7%) followed by swelling (26.8%), ulcer (26.2%). Positive and negative predictive value of TB was 91.8% and 86.5% respectively whereas positive and negative predictive value of histopathology was 96.4% and 72.7% respectively. TB stain is cheap, quick and easily available readymade kit and no side effects and so have excellent patient compliance so it can be best utilized as a premier tool in initial detection of suspected oral malignancy.


The fetal three-vessel and tracheal view revisited.
The routine use of four-chamber screening of the fetal heart was pioneered in the early 1980s and has been shown to detect reliably mainly univentricular hearts in the fetus. Many conotruncal anomalies and ductal-dependent lesions may, however, not be detected with the four-chamber view alone and additional planes are needed. The three-vessel and tracheal (3VT) view is a transverse plane in the upper mediastinum demonstrating simultaneously the course and the connection of both the aortic and ductal arches, their relationship to the trachea and the visualization of the superior vena cava. The purpose of the article is to review the two-dimensional anatomy of this plane and the contribution of colour Doppler and to present a checklist to be achieved on screening ultrasound. Typical suspicions include the detection of abnormal vessel number, abnormal vessel size, abnormal course and alignment and abnormal colour Doppler pattern. Anomalies such as pulmonary and aortic stenosis and atresia, aortic coarctation, interrupted arch, tetralogy of Fallot, common arterial trunk, transposition of the great arteries, right aortic arch, double aortic arch, aberrant right subclavian artery, left superior vena cava are some of the anomalies showing an abnormal 3VT image. Recent studies on the comprehensive evaluation of the 3VT view and adjacent planes have shown the potential of visualizing the thymus and the left brachiocephalic vein during fetal echocardiography and in detecting additional rare conditions. National and international societies are increasingly recommending the use of this plane during routine ultrasound in order to improve prenatal detection rates of critical cardiac defects.


Contrasting patterns of autonomic dysfunction in patients with mitral valve prolapse and panic attacks.
Both mitral valve prolapse (MVP) and panic attacks have been reported to be associated with autonomic dysfunction, but previous studies have been limited by the lack of clear separation between patients with MVP and those with panic attacks and the use of noncomparable control subjects. Accordingly, heart rate and blood pressure responses to deep breathing, five minutes' quiet standing, and the Valsalva maneuver were studied in age- and cardiac symptom-matched groups of 33 control subjects, 66 patients with MVP, 20 patients with panic attacks, and 17 patients with both MVP and panic attacks. Compared with control subjects, patients with MVP exhibited more syncope (13 of 66, or 20 percent, versus none of 33, or 0 percent; p less than 0.01), more orthostatic hypotension during quiet standing (11 of 66, or 17 percent, versus one of 33, or 3 percent; p less than 0.01), loss of the normal decrease with age in vagally-mediated heart rate variability during deep breathing (r = 0.13, p = NS versus r = -0.44, p = 0.01), and lower 24-hour epinephrine excretion (6.1 +/- 0.7 versus 11.0 +/- 2.7 micrograms; p less than 0.01). In contrast, patients with panic attacks had greater increases than control subjects or patients with MVP without panic attacks in heart rate, mean blood pressure, and the product of heart rate and mean blood pressure during each minute of quiet standing and during the early strain phase of the Valsalva maneuver. These findings indicate that autonomic dysfunction occurs both in patients with MVP and in those with panic attacks in comparison with symptomatic control subjects, but that the patterns of abnormality differ. Patients with MVP exhibit decreased effectiveness of responses to orthostatic stress, reduced epinephrine excretion, and abnormal vagal tone, whereas patients with panic attacks have heightened cardiovascular responses to postural and positive intrathoracic pressure stresses.


Molecular signatures of neoadjuvant endocrine therapy for breast cancer: characteristics of response or intrinsic resistance.
Approximately 30% of patients with estrogen receptor (ER) positive breast cancers exhibit de novo or intrinsic resistance to endocrine therapies. The purpose of this study was to define genes that distinguish ER+ resistant from ER+ responsive tumors, prior to the start of hormone therapies. Previously untreated post-menopausal patients with ER+ breast cancers were treated for 4 months in a neoadjuvant setting with the aromatase inhibitor exemestane alone, or in combination with the antiestrogen tamoxifen. Matched pre- and post-treatment tumor samples from the same patient, were analyzed by gene expression profiling and were correlated with response to treatment. Genes associated with tumor shrinkage achieved by estrogen blockade therapy were identified, as were genes associated with resistance to treatment. Prediction Analysis of Microarrays (PAM) identified 50 genes that can predict response or intrinsic resistance to neoadjuvant endocrine therapy of ER+ tumors, 8 of which have been previously implicated as useful biomarkers in breast cancer. In summary, we identify genes associated with response to endocrine therapy that may distinguish ER+, hormone responsive breast cancers, from ER+ tumors that exhibit intrinsic or de novo resistance. We suggest that the estrogen signaling pathway is aberrant in ER+ tumors with intrinsic resistance. Lastly, the studies show upregulation of a "lipogenic pathway" in non-responsive ER+ tumors that may serve as a marker of intrinsic resistance. This pathway may represent an alternative target for therapeutic intervention.


Distinct roles of TNF-related apoptosis-inducing ligand (TRAIL) in viral and bacterial infections: from pathogenesis to pathogen clearance.
Apoptotic death of different cells observed during infection is thought to limit overwhelming inflammation in response to microbial challenge. However, the underlying apoptotic death mechanisms have not been well defined. Tumor necrosis factor (TNF) related apoptosis-inducing ligand (TRAIL) is a type II transmembrane protein belonging to the TNF superfamily, which is involved not only in tumor growth suppression but in infection control and also in the regulation of both innate and adaptive immune responses. In this review, we have summarized data of recent studies on the influence of the TRAIL/TRAIL receptor (TRAIL-R) system on the development of viral and bacterial infections. TRAIL may have a dual function in the immune system being able to kill infected cells and also to participate in the pathogenesis of multiple infections. Moreover, many pathogens have evolved mechanisms to manipulate TRAIL signaling thus increasing pathogen replication. Present data highlight an essential role for the TRAIL/TRAIL-R system in the regulation and modulation of apoptosis and show that TRAIL has distinct roles in pathogenesis and pathogen elimination. Knowledge of the factors that determine whether TRAIL is helpful or harmful supposes its potential therapeutic implications that are only beginning to be explored.


Shear Failure in Supported Two-Dimensional Nanosheet Van der Waals Thin Films.
Liquid-phase deposition of exfoliated 2D nanosheets is the basis for emerging technologies that include writable electronic inks, molecular barriers, selective membranes, and protective coatings against fouling or corrosion. These nanosheet thin films have complex internal structures that are discontinuous assemblies of irregularly tiled micron-scale sheets held together by van der Waals (vdW) forces. On stiff substrates, nanosheet vdW films are stable to many common stresses, but can fail by internal delamination under shear stress associated with handling or abrasion. This "re-exfoliation" pathway is an intrinsic feature of stacked vdW films and can limit nanosheet-based technologies. Here we investigate the shear stability of graphene oxide and MoSe2 nanosheet vdW films through lap shear experiments on polymer-nanosheet-polymer laminates. These sandwich laminate structures fail in mixed cohesive and interfacial mode with critical shear forces from 40 - 140 kPa and fracture energies ranging from 0.2 - 6 J/m2. Surprisingly these energies are higher than delamination energies reported for smooth peeling of ordered stacks of continuous 2D sheets, which we propose is due to energy dissipation and chaotic crack motion during nanosheet film disassembly at the crack tip. Experiment results also show that film thickness plays a key role in determining critical shear force (maximum load before failure) and dissipated energy for different nanosheet vdW films. Using a mechanical model with an edge crack in the thin nanosheet film, we propose a shear-to-tensile failure mode transition to explain a maximum in critical shear force for graphene oxide films but not MoSe2 films. This transition reflects a weakening of the substrate confinement effect and increasing rotational deformation near the film edge as the film thickness increases. For graphene oxide, the critical shear force can be increased by electrostatic cross-linking achieved through interlayer incorporation of metal cations. These results have important implications for the stability of functional devices that employ 2D nanosheet coatings.


Correlations between the levels of acute infection markers and serum albumin in elderly patients with hip fracture.
The aim of this study was to explore a clinical index that could predict the decline of serum albumin (ALB) in elderly patients (over 60 years old) with hip fractures in 2014. All the data came from the retrospective survey, and the correlations between the ALB changes and acute infection markers were then analyzed using correlation analysis. The changes of infection markers and ALB before and after surgery were compared using the t test. There was no correlation of the serum ALB blood with interleukin-6 (IL-6) (r = 0.072, P = 0.588), C-reactive protein (CRP) (r = -0.249, P = 0.057), or calcitonin (PCT) (r = -0.038, P = 0.775) when patients were admitted, but it was negatively correlated with the total amount of infection markers (TAIMs) (r = -0.301, P = 0.020). The postoperative levels of IL-6 (154.23 ± 177.14 pg/mL) (P < 0.001), CRP (69.52 ± 39.84 mg/L) (P < 0.001), and PCT (1.27 ± 2.4 ng/mL) (P < 0.001) were significantly increased than those before surgery [IL-6 (44.96 ± 54.58 pg/mL), CRP (31.78 ± 29.90 mg/L), and PCT (0.42 ± 1.06 ng/mL)]. The postoperative level of serum ALB (29.93 ± 3.02 g/L) was significantly reduced than that before surgery (33.95 ± 3.69 g/L) (P < 0.001). The serum ALB level was negatively correlated with IL-6 (r = -0.333, P = 0.015) before surgery, but not correlated with TAIMs (r = -0.256, P = 0.061). The serum ALB level was negatively correlated with IL-6 (r = -0.292, P = 0.034) and TAIMs (r = -0.271, P = 0.050) after surgery. The serum IL-6 level could predict the changes of ALB during the disease process.


An extended U2AF(65)-RNA-binding domain recognizes the 3' splice site signal.
How the essential pre-mRNA splicing factor U2AF(65) recognizes the polypyrimidine (Py) signals of the major class of 3' splice sites in human gene transcripts remains incompletely understood. We determined four structures of an extended U2AF(65)-RNA-binding domain bound to Py-tract oligonucleotides at resolutions between 2.0 and 1.5 Å. These structures together with RNA binding and splicing assays reveal unforeseen roles for U2AF(65) inter-domain residues in recognizing a contiguous, nine-nucleotide Py tract. The U2AF(65) linker residues between the dual RNA recognition motifs (RRMs) recognize the central nucleotide, whereas the N- and C-terminal RRM extensions recognize the 3' terminus and third nucleotide. Single-molecule FRET experiments suggest that conformational selection and induced fit of the U2AF(65) RRMs are complementary mechanisms for Py-tract association. Altogether, these results advance the mechanistic understanding of molecular recognition for a major class of splice site signals.


Early results of bladder-cancer screening in a high-risk population of heavy smokers.
To report first results of an early bladder-cancer detection programme, and to evaluate the detection rate and the diagnostic value of the tests used. Urine samples of 183 screened subjects with a history of smoking of > or =40 pack-years were collected for analysis with a urinary dipstick test for haematuria, the nuclear matrix protein-22 test (BladderChek, Matritech, Inc., Newton, MA, USA), voided urine cytology and a molecular cytology test (UroVysion, Abbott Molecular Inc., Des Plaines, IL, USA). Participants with at least one positive test result had a further evaluation including cystoscopy and radiological imaging. The subjects' risk factors, test results and histological findings were analysed. In all, 75 subjects had at least one positive test result and were evaluated further; abnormal histological findings were detected in 18 (24% of those who had cystoscopy, 9.8% of the original 183), 15 of those in the urinary bladder, with pTaG1 (one), carcinoma in situ (two), dysplastic lesions (11) and one an inverted papilloma. In the upper urinary tract, two urothelial tumours (pTaG1 and pTxN2G3) and one renal cell carcinoma (pT1G2) were detected by computed tomography. In summary, six of 183 subjects (3.3%) had a histologically confirmed malignant tumour and another 12 (6.6%) were identified with a possible pre-cancerous lesion of the urinary tract. The urinary dipstick, BladderChek, cytology and UroVysion detected (i.e. were true-positive in) nine (50%), one (6%), seven (39%) and 11 (61%) of the 18 tumours found, while they failed to detect nine (50%), 17 (94%), 11 (61%) and seven (39%) of these lesions, respectively. Omitting the urine dipstick test, the BladderChek, cytology or UroVysion from the test setting could have spared 40, five, two or one subjects(s) from unnecessary invasive interventions; however, three, none, two or six lesions, would have been missed. More positive screening tests per subject was associated with a higher probability of a (pre)-malignant lesion. Screening a high-risk group with a history of smoking of > or =40 pack-years showed a significant proportion (3.3%) with malignancy. These first results are encouraging and warrant continuation of the screening programme. In this series the most efficient screening tool was the combination of UroVysion, cytology and urinary dipstick testing. Of special scientific interest will be the follow-up of those patients with a possible pre-cancerous lesion.


Pathological Features of Persistent Atrial Standstill Syndrome in Three Dogs.
The hearts of three dogs, clinically diagnosed as having persistent atrial standstill syndrome (PAS), were studied post mortem. The most significant gross findings in the hearts of all three dogs were dilatation and marked reduction in the thickness of both atrial walls. Histopathologically, all three had widespread progressive loss of the atrial myocardium with replacement by fatty or fibrofatty tissue, consistent with atrial myopathy. The lesion mainly affected the upper half of both atria and was more severe in the epimyocardium and midmyocardium than in the endomyocardium. On the basis of these observations, it is proposed that the atrial myopathy commences in the upper regions of both atria and progresses downwards, as has been demonstrated electrophysiologically in PAS in humans, and extends from the epicardium towards the endocardium.


Electrophoretic transfer of viral proteins to nitrocellulose sheets and detection with peroxidase-bound lectins and protein A.
Peroxidase-bound protein A and lectins in combination with precipitable substrates were used to detect viral proteins and glycoproteins on 'Western blots'. The sensitivity of various methods was compared, including radioiodinated protein A, enzyme immunosorbent assay and peroxidase staining with DAP. The sensitivity of the peroxidase reaction, its rapidity and low cost compared with other methods make this a useful choice for developing protein bands transferred to nitrocellulose paper. In addition, a procedure was devised to clarify and fix nitrocellulose sheets for direct quantification of the precipitated substrate by densitometric analysis.


Comparison of the severity and outcome of invasive pneumococcal infections in children and adults.
In a population-based observational study of 285 patients with positive blood culture for pneumococci, we found that the course and outcome of invasive pneumococcal infections differ considerably between adults and children. None of the children died, whereas the infection was fatal for 15% (35/229) of the adults (P<0.001). The differences are only partly explained by the underlying conditions.


Molecular characteristics and alterations during early development of the human vagina.
Unresolved questions remain concerning the derivation of the vagina with respect to the relative contributions from the Müllerian ducts, the urogenital sinus, and the Wolffian ducts. Recent molecular and cellular studies in rodents have opened up a large gap between the level of understanding of vaginal development in mice and understanding of human vaginal development, which is based on histology. To compare the findings in mice with human vaginal development and to address this gap, we analysed molecular characteristics of the urogenital sinus, Wolffian ducts, and Müllerian ducts in 8-14-week-old human specimens using immunohistochemical methods. The monoclonal antibodies used were directed against cytokeratin (CK) 14, CK19, vimentin, laminin, p63, E-cadherin, caspase-3, Ki67, HOX A13, and BMP-4. The immunohistochemical analysis revealed that, during weeks 8-9, the epithelium of the Müllerian ducts became positive for p63 as p63-positive cells that originated from the sinus epithelium reached the caudal tip of the fused Müllerian ducts via the Wolffian ducts. The lumen of the fused Müllerian ducts was closed by an epithelial plug that contained both vimentin-positive and vimentin-negative cells. Subsequently, the resulting epithelial tube enlarged by proliferation of basal p63-positive cells. The first signs of squamous differentiation were detected during week 14, with the appearance of CK14-positive cells. According to our results, all three components, namely, the urogenital sinus, Wolffian ducts, and Müllerian ducts, interacted during the formation of the human vagina. The sinus epithelium provided p63-positive cells, the Wollfian ducts acted as a 'transporter', and the Müllerian ducts contributed the guiding structure for the vaginal anlagen. Epithelial differentiation began at the end of the period studied and extended in a caudo-cranial direction. The present study is one of the first to provide up-to-date molecular correlates for human vaginal development that can be compared with the results of cell biological studies in rodents.


Quality of life and mental health in amblyopic children treated with and without occlusion therapy.
To assess the long-term (≥1 year) psychosocial effects of occlusion therapy in children with refractive amblyopia and their caregivers. Multiple self-reported questionnaires were used to investigate HRQoL, anxiety, depression, and stress in a cross-sectional study of children with refractive amblyopia and their caregivers. Children were divided into two subgroups: those treated with glasses only and those treated with both glasses and occlusion therapy who had stopped wearing a patch at least 1 year earlier. A total of 79 children were included: 32 in the refraction-only group and 47 in the glasses + occlusion group. Median age of subjects was 12 years (IQR = 3.0). Mean age of the 79 caregivers was 41.9 ± 5.0 years. No significant differences were found for any of the psychosocial outcomes analyzed between children treated with glasses and patching and those treated with glasses only. The differences remained insignificant between children according to age of first occlusion (<6 vs ≥6 years). The only significant (but not clinically relevant) difference observed for caregivers was a higher score in the depression dimension of the Hospital Anxiety and Depression Scale for caregivers of children treated with glasses only (P = 0.036; effect size = 0.23). In our study cohort, patching was not associated with long-term negative psychosocial outcomes in children with refractive amblyopia treated with occlusion therapy.


Impact of a program for the prevention of traffic accidents in a Southern Brazilian city: a model for implementation in a developing country.
Traffic accidents constitute the main cause of death in the first decades of life. Traumatic brain injury is the event most responsible for the severity of these accidents. The SBN started an educational program for the prevention of traffic accidents, adapted from the American model "Think First" to the Brazilian environment, since 1995, with special effort devoted to the prevention of TBI by using seat belts and motorcycle helmets. The objective of the present study was to set up a traffic accident prevention program based on the adapted Think First and to evaluate its impact by comparing epidemiological variables before and after the beginning of the program. The program was executed in Maringá city, from September 2004 to August 2005, with educational actions targeting the entire population, especially teenagers and young adults. The program was implemented by building a network of information facilitators and multipliers inside the organized civil society, with widespread population dissemination. To measure the impact of the program, a specific software was developed for the storage and processing of the epidemiological variables. The results showed a reduction of trauma severity due to traffic accidents after the execution of the program, mainly TBI. The adapted Think First was systematically implemented and its impact measured for the first time in Brazil, revealing the usefulness of the program for reducing trauma and TBI severity in traffic accidents through public education and representing a standardized model of implementation in a developing country.


A new method for noninvasive measurement of pulmonary gas exchange using expired gas.
Measurement of the gas exchange efficiency of the lung is often required in the practice of pulmonary medicine and in other settings. The traditional standard is the values of the PO2, PCO2, and pH of arterial blood. However arterial puncture requires technical expertise, is invasive, uncomfortable for the patient, and expensive. Here we describe how the composition of expired gas can be used in conjunction with pulse oximetry to obtain useful measures of gas exchange efficiency. The new procedure is noninvasive, well tolerated by the patient, and takes only a few minutes. It could be particularly useful when repeated measurements of pulmonary gas exchange are required. One product of the procedure is the difference between the PO2 of end-tidal alveolar gas and the calculated PO2 of arterial blood. This measurement is related to the classical alveolar-arterial PO2 difference based on ideal alveolar gas. However that traditional index is heavily influenced by lung units with low ventilation-perfusion ratios, whereas the new index has a broader physiological basis because it includes contributions from the whole lung.


A possible new HLA-DR allele.
In routine screening of anti-HLA DR reagents, serum 901 was obtained from a woman of negroid origin ten days after delivery of a second child. The spouse was also of black origin. This serum contained polyspecific HLA A and B antibodies. After platelet absorption it reacted with the B cells of 10 out of 119 European Caucasoid panel donors (8.4%) typed for DR1 to DRW10 and for MT1, MT2. Each of these ten donors had only one recognized DR antigen, the other was "blank." Serum 901 gave negative reactions with the homozygous typing cells (HTC) DW1 to DW8. In 18 normal informative Caucasoid families, serum 901 recognized one HLA haplotype in one (or both) parents and segregated with this haplotype in one or more than one child. In one family in which both parents reacted with serum 901, two children were homozygous for this marker and reacted as HTCs. One of these HTCs typed as DW9 and was found to be identical to a DW9.HTC (8W207).


A clinical evaluation of isotope scanning, ultrasonography and computed tomography in pancreatic disease.
In a prospective study of 46 patients with suspected pancreatic disease the provisional diagnoses arrived at independently by isotope scanning (IS), ultrasonography (USS) and computed tomography (CT) have been compared. In the control group, IS and CT were associated with a higher false positive rate than USS; The isotope scan was abnormal in most patients with proven chronic pancreatitis and cancer. The results from USS and CT were similar when structural changes were present. USS was superior in diagnosing pancreatic carcinoma and was a convenient means to follow the progression of acute pancreatitis to final resolution or the development of a pseudocyst. CT proved especially useful in accurately delineating cysts, pseudocysts and calculi prior to planning surgery and in assessing disease in contiguous viscera.


Photodegradation of Rhodamine B over unexcited semiconductor compounds of BiOCl and BiOBr.
This study reported, for the first time systematically, photodegradation of Rhodamine B (RhB) in aqueous solution over BiOCl and BiOBr semiconductors. Under visible light irradiation (λ>400 nm, λ>420 nm and λ=550±15 nm), RhB adsorbed on the surface of BiOCl and BiOBr was photosensitized and decomposed effectively over unexcited BiOCl and BiOBr. The degradation of Methyl Orange (MO) and Methylene Blue (MB) over BiOCl and BiOBr was investigated as well, and the results were compared with RhB photodegradation. It was found that MB molecules having the lowest LUMO could not be degraded by this process. Utilizing the quantum chemical calculation (Gaussian 03 program), the relationship between frontier orbital energy of selected dye molecules and photodegradation rate was established for the first time in this study.


Paramagnetic effects in NMR for protein structures and ensembles: Studies of metalloproteins.
Paramagnetic effects on the NMR spectra are known to encode information on structure, electronic properties and dynamics hardly accessible with any other technique, especially in the field of biological systems. Paramagnetism-based restraints are conveniently used for the de novo determination of protein structures, the structural refinement starting from crystallographic models, and for the determination of the internal arrangement of domains with known structures. Conformational variability can also be profitably interrogated including the possibility of uncovering the presence of states with very low population. The recent advances in the quantum chemistry treatment of paramagnetic NMR effects has provided new momentum to the field, allowing for the refinement of protein structures at the metal coordination site to an unprecedented resolution.


Birth-related mid-posterior rib fractures in neonates: a report of three cases (and a possible fourth case) and a review of the literature.
Posterior rib fractures in young children have a high positive predictive value for non-accidental injury (NAI). Combined data of five studies on birth trauma (115,756 live births) showed no cases of rib fractures resulting from birth trauma. There have, however, been sporadic cases reported in the literature. We present three neonates with both posterior rib fractures and ipsilateral clavicular fractures resulting from birth trauma. A review of the literature is also presented. The common denominator and a possible mechanical aetiology are discussed. In total, 13 cases of definitive birth-related posterior rib fractures were identified. Nearly all (9/10) posterior rib fractures were (as far as reported in the original publications) in the midline. In 12 of the 13 children, birth weight was high and in 7 children birth was complicated by shoulder dystocia. An interesting finding was that in cases where a clavicular fracture was present, this was on the ipsilateral side. Radiologists, when presented with a neonate with posterior rib fractures, should be aware of this rare differential diagnosis.


Partial scapholunate ligament injuries treated with arthroscopic debridement and thermal shrinkage.
To present the early results of arthroscopic debridement and thermal shrinkage using radiofrequency probes for partial (Geissler grades I and II) scapholunate (SL) interosseous ligament injuries of the wrist. Sixteen patients with a mean age of 34 years (range, 18-54 y) presenting with chronic dorsoradial wrist pain unresponsive to initial conservative treatment for a mean period of 12 weeks were included in this study. No patient showed radiologic signs of static dissociation (SL interval, <3.5 mm; mean SL angle, 49 degrees ) before surgery. Diagnostic arthroscopy showed a partial SL tear in 14 patients and redundancy of the ligament in 2. Partial SL tears involved the membranous (proximal) and volar part of the ligament. All lesions were debrided and treated with thermal shrinkage using a bipolar radiofrequency probe. The mean follow-up period was 19 months (range, 9-34 mo). Fourteen patients experienced substantial pain relief whereas in 2 the pain remained unchanged. Eight patients were completely pain free. The mean flexion-extension arc was 142 degrees and the mean grip strength was 78% that of the unaffected side. No patient showed radiologic signs of arthritis or static or dynamic instability after surgery (SL interval remained <3.5 mm; mean SL angle, 53 degrees ). Based on the modified Mayo wrist score there were 8 excellent, 6 good, 1 fair, and 1 poor result. Partial SL ligament tears can be a source of radial-sided wrist pain. Scapholunate ligament debridement and thermal shrinkage effectively provided pain relief for most of the patients treated. Stability was maintained radiographically. No complications were noted from the use of radiofrequency probes. These reasonably favorable short-term results should be viewed cautiously. A longer follow-up study is necessary to determine the ultimate efficacy of this procedure.


Cost-Effectiveness Analysis of Biological Signature DCISionRT Use for DCIS Treatment.
Currently it remains difficult to identify patients most likely to benefit from radiotherapy (RT) for ductal carcinoma-in-situ (DCIS), thus leading to wide variation in practice patterns. The genomic risk assessment tool DCISionRT (PreludeDX) has been validated to prognosticate recurrence risk and predict RT benefit. We aimed to study the cost-effectiveness analysis comparing DCIS treatments based on DCISionRT testing to traditional clinicopathologic risk factors. A Markov state transition model was constructed to perform a cost-effectiveness analysis comparing breast-conserving surgery with or without RT using DCISionRT testing vs. traditional clinicopathologic risk factors. Clinical parameters were obtained from clinical trial data and cross-validation studies. Cost data were based on 2019 Medicare reimbursement. Incremental cost-effectiveness ratio (ICER) was calculated as incremental cost per quality-adjusted life-year (QALY) gained comparing DCIS treatments using DCISionRT testing to traditional clinicopathologic risk factors and evaluated with a willingness-to-pay threshold of US$100,000 per QALY gained. To account for uncertainty, 1-way and probabilistic sensitivity analyses were performed. Base case analysis showed that DCIS management using DCISionRT testing was a cost-effective strategy, resulting in an ICER of $74,331 per QALY gained compared to clinicopathology-based treatment. Model results were sensitive to a variation of the proportion of genomic-high, low-risk patients receiving RT in DCISionRT testing strategy, and changes in DCISionRT testing cost. DCISionRT testing could potentially be a cost-effective strategy compared to traditional decision making for DCIS treatments, optimizing RT benefit based on an accurate recurrence risk assessment.


Effects of the Intraneural and Subparaneural Ultrasound-Guided Popliteal Sciatic Nerve Block: A Prospective, Randomized, Double-Blind Clinical and Electrophysiological Comparison.
This prospective, randomized, double-blind study compared the effects of the ultrasound-guided popliteal sciatic nerve block performed by either intraneural or subparaneural approach followed by an electrophysiological evaluation. We hypothesized that intraneural injection provides a faster onset with a better success rate compared with the subparaneural approach. Eighty-eight patients were enrolled and randomized to receive an ultrasound-guided popliteal sciatic nerve block injecting 15 mL ropivacaine 1% according to an intraneural injection (group INTRA = 44) or a subparaneural injection (group SUBPARA = 44). The primary end point was the onset time of sensory and motor block, whereas secondary end points were successful, duration of the block, and the variation of the electrophysiological assessment after 5 weeks. The study was registered prior to patient enrollment (clinicaltrials.gov identifier NCT01987128). The median onset time for successful sciatic nerve block in the INTRA group was 10 (5-15 [5-30]) minutes versus 25 (15-35 [5-45]) minutes in the SUBPARA group (P < 0.001), with a rate of 41/43 (95.3%) compared with 25/40 (62.5%) in the SUBPARA group (P < 0.001). No difference was found regardless of the duration of the block. Fifty-three patients also performed the electrophysiological assessment at 5 weeks, showing a subclinical, significant reduction in amplitude of the action potentials with no difference between groups. No patients reported any clinical neurological complications after 6 months. In ultrasound-guided popliteal sciatic nerve block, intraneural injection provided a faster onset and better success rate compared with subparaneural. Both techniques resulted in a similar subclinical reduction in amplitude of the sciatic action potentials at 5 weeks after surgery. These findings should not be extended to other approaches.


Chronic hypoparathyroidism is associated with increased cortical bone density evaluated using high-resolution peripheral quantitative computed tomography.
This cross-sectional study aimed to assess bone mineral density (BMD), bone microarchitecture and fracture prevalence in women with chronic postsurgical hypoparathyroidism (hypoPT). Twenty-seven women with postsurgical hypoPT and 44 age-matched healthy women were included. Dual-energy X-ray absorptiometry was used to evaluate areal BMD and vertebral fracture assessment. High-resolution peripheral quantitative computed tomography assessed microarchitecture and volumetric BMD at the distal radius and tibia. Biochemical parameters, including fibroblast growth factor 23, C-terminal cross-linking telopeptide of type I collagen (ICTP), and procollagen type I N-terminal propeptide (P1NP), were also measured. Previous low-impact fractures were assessed and the 10-year fracture risk was estimated using the FRAX tool for the Brazilian population. No participant had prevalent clinical fractures, and both groups showed low risk for major and hip based on FRAX tool, but two hypoPT patients had moderate to severe morphometric vertebral fractures. Women with hypoPT had increased aBMD in the lumbar spine, femoral neck and total hip (p < 0.05) and higher cortical vBMD in the radius (p = 0.020) and tibia (p < 0.001). Trabecular bone was not affected. Both P1NP and ICTP suggested low bone turnover rates, but no significant correlation was observed between bone density or microstructure and any of the biochemical parameters. The prevalence of fragility fractures was low in HypoPT women and compatible with low fracture risk estimated by the FRAX tool. Patients had a higher aBMD and cortical vBMD than those of healthy control women, but the association with decreased bone turnover remains unclear.


Complications of the Use of the OverStitch Endoscopic Suturing System.
Endoscopic suturing with the OverStitch device is safe and effective for a wide range of applications from defect closure and stent fixation to hemostasis and bariatrics. OverStitch-related complications remain rare, although adverse events such as bleeding, mucosal injury, perigastric fluid collections, leaks, and perforations have been reported. Provider familiarity with the device and the specific pitfalls that may arise with OverStitch use in different situations is necessary to minimize the risk of adverse events. This article reviews potential complications and provides tips and troubleshooting techniques.


Vimentin expression in spermatogenic and Sertoli cells is stage-related in rat seminiferous epithelium.
Monoclonal and polyclonal antibodies were used in indirect immunofluorescence microscopy to localize vimentin intermediate filaments in the rat seminiferous epithelium. During stages XII-V of the epithelial cycle, the Sertoli cells showed a reaction in the perinuclear area and vimentin-positive extensions, projecting toward the developing spermatid bundles, were also seen. During stages VI-XI these extensions were small and narrow. Monoclonal antibody to vimentin gave a granular reaction in the peripheral region of the flagella of steps 16-19 spermatids. Western blotting indicated a specific reaction with a Mr 58,000 polypeptide in isolated seminiferous tubules and in epididymal spermatozoa. Our results suggest that vimentin filaments in Sertoli cells may be regulated cyclically in a stage-dependent manner. The granular reaction in the spermatid flagellum with the monoclonal antibody suggests that vimentin in germ cells is organized differently from that in somatic Sertoli cells.


Efficacy and safety of methotrexate in recalcitrant cutaneous lupus erythematosus: results of a retrospective study in 43 patients.
The therapy of cutaneous lupus erythematosus (CLE) is often challenging, especially in patients resistant to topical treatment and established first-line systemic drugs such as antimalarials. Systemic corticosteroids are effective, but their use is limited due to well-known side-effects, especially in long-term treatment. In recent years several other immunosuppressive agents have been successfully applied in CLE. However, there are no large studies or explicit guidelines on the use of these drugs in CLE. To perform a retrospective investigation of the efficacy of low-dose methotrexate (MTX) in the treatment of CLE. One hundred and thirty-nine patients with CLE were seen at our department between 2001 and 2003, of whom 43 patients required low-dose MTX. All had histologically confirmed CLE lesions. Clinical data including disease activity, additional treatment, laboratory parameters and side-effects were recorded carefully at the time of presentation. Statistical analyses were performed by paired nonparametric Wilcoxon test and Student's t-test using SPSS 11 software. MTX led to a highly significant (P < 0.01) decline in disease activity. An improvement of the cutaneous lesions was recorded in nearly all patients treated with MTX (42 of 43; 98%). Severe side-effects necessitating discontinuation of MTX treatment were recorded in seven patients (16%), which quickly resolved when MTX was discontinued. Life-threatening complications were not observed. Intravenous application was tolerated better than oral administration. Interestingly, we observed a significant increase in circulating lymphocyte numbers in patients with lymphopenia (< 1.0 x 10(9) cells L(-1)) prior to MTX treatment. Our study supports earlier findings reporting the efficacy of low-dose MTX in CLE lesions, particularly in recalcitrant clinical courses. MTX treatment appears to be safe if patients are carefully selected and monitored, with particular attention to side-effects and contraindications.


Sporozoite and liver stage antigen Plasmodium falciparum peptides bind specifically to human hepatocytes.
Sporozoite and Liver Stage Antigen (SALSA) sequence synthetic peptides were used in HepG2 cell binding assays to identify regions involved in parasite invasion. SALSA 20608 ( 21IWASEKKDEKEASEQGEESHY40) and 20611 ( 64KKDDGTDKVQEKVLEKSPKY83) peptides were determined as having high binding activity in HepG2 cell assays, some of them were located in immunogenic regions. Immune-fluorescence antibody test with 24276 (20608 peptide analogue, CGIWSSMKMDEKMAAMQGEESHCG) showed sporozoite and merozoite reactivity. This data suggests SALSA high activity binding peptides' (HABPs) possible role in hepatic cell invasion and merozoite invasion of erythrocytes.


Clinical management of lung cancer patients with respiratory symptoms during the COVID-19 pandemic.
There are many clinical conditions, such as lung cancer, that need to be followed up and treated during a pandemic. Providing health care for patients who are immune-suppressive requires extra care. Among 108 lung cancer patients who had been hospitalized during the COVID-19 pandemic, 18 with respiratory symptoms were evaluated retrospectively. The patients' median age was 64 ± 9.4 with a male predominance (male n = 16, female n = 2). Thirteen had non-small cell lung cancer (NSCLC), and 5 had small cell lung cancer (SCLC). Nine (50%) patients were receiving chemotherapy. The most common symptom was shortness of breath (n = 14, 77.8%), followed by fever (n = 10, 55.6%). The findings confirmed on computed thorax tomography (CTT) were as follows: consolidation (n = 8, 44.4%), ground glass opacities (n = 8, 44.4%) and thoracic tumour/mediastinal-hilar lymphadenopathy (n = 3, 16.7%). Hypoxia was seen in 11 patients (61.1%), twelve patients had an elevated LDH (median = 302 ± 197) and lymphopenia (median = 1055 ± 648) and 5 (27.7%) were highly suspected of having contracted COVID-19. None of their nasopharyngeal swaps was positive. Two of these 5 patients received COVID-19 specific treatment even though they thrice had negative reverse transcription polymerase chain reaction (RT-PCR) results. The two patients responded well to both clinical and radiological treatments. For one case with SCLC receiving immunotherapy, methylprednisolone was initiated for radiation pneumonitis after excluding COVID-19. In line with a country's health policies and the adequacy of its health system, the necessity of a multidisciplinary approach in the management and treatment of complications in patients with lung cancer has become even more important during the COVID-19 pandemic.


Gene transfer of Fc-fusion cytokine by in vivo electroporation: application to gene therapy for viral myocarditis.
Among a number of techniques for gene transfer in vivo, the direct injection of plasmid DNA into muscle is simple, inexpensive and safe. Although combining direct DNA injection with in vivo electroporation increases the efficiency of gene transfer into muscle, applications of this method have remained limited because of the relatively low expression level. To overcome this problem, we developed a plasmid vector that expresses a secretory protein as a fusion protein with the noncytolytic immunoglobulin Fc portion and used it for electroporation-mediated viral interleukin 10 (vIL-10) expression in vivo. The fusion cytokine vIL-10/mutFc was successfully expressed and the peak serum concentration of vIL-10 was almost 100-fold (195 ng/ml) higher than with a non-fusion vIL-10 expression plasmid. The expressed fusion cytokine suppressed the phytohemagglutinin-induced IFN-gamma production by human peripheral blood mononuclear cells and decreased the mortality in a mouse viral myocarditis model as effectively as vIL-10 expression. These results demonstrate that the transfer of plasmid DNA expressing a noncytolytic Fc-fusion cytokine is useful to deliver enhanced levels of cytokine without altering general biological activities. This simple and efficient system should provide a new approach to gene therapy for human diseases and prove very useful for investigating the function of newly discovered secretory protein genes.


Speech perception enhancement in elderly hearing aid users using an auditory training program for mobile devices.
The goal of the present study was to develop an auditory training program using a mobile device and to test its efficacy by applying it to older adults suffering from moderate-to-severe sensorineural hearing loss. Among the 20 elderly hearing-impaired listeners who participated, 10 were randomly assigned to a training group (TG) and 10 were assigned to a non-training group (NTG) as a control. As a baseline, all participants were measured by vowel, consonant and sentence tests. In the experiment, the TG had been trained for 4 weeks using a mobile program, which had four levels and consisted of 10 Korean nonsense syllables, with each level completed in 1 week. In contrast, traditional auditory training had been provided for the NTG during the same period. To evaluate whether a training effect was achieved, the two groups also carried out the same tests as the baseline after completing the experiment. The results showed that performance on the consonant and sentence tests in the TG was significantly increased compared with that of the NTG. Also, improved scores of speech perception were retained at 2 weeks after the training was completed. However, vowel scores were not changed after the 4-week training in both the TG and the NTG. This result pattern suggests that a moderate amount of auditory training using the mobile device with cost-effective and minimal supervision is useful when it is used to improve the speech understanding of older adults with hearing loss. Geriatr Gerontol Int 2017; 17: 61-68.


Chronic antidepressant treatment and central 5-HT synapses.
The present studies have shown that chronic antidepressant treatment with desipramine, imipramine, zimelidine and alaproclate led to adaptive changes in both pre- and postsynaptic 5-hydroxytryptamine (5-HT) receptor mechanisms which appear to result in 5-HT sub- or supersensitivity development depending upon the 5-HT nerve terminal systems analyzed. The results underline the heterogeneity of the central 5-HT neurone systems in their responses to chronic antidepressant treatment. An involvement of 5-HT comodulators is postulated in the actions of antidepressants on central 5-HT synapses. Finally, the results point to the importance of 5-HT neurone systems as targets for the action for antidepressant drugs and they may mediate at least some of the therapeutic activity of antidepressant drugs.


Arthroscopic Remnant Preserving 3-Portal Posterior Cruciate Ligament Reconstruction With Reverse Passage of the Graft.
The posterior cruciate ligament (PCL) is the largest and strongest ligament of the knee, with a tremendous role as the primary posterior stabilizer of the knee. Surgical management of PCL injuries is very demanding, due to the fact that the PCL tear usually is a part of multiligamentous knee injury. Moreover, PCL anatomy, especially its course and attachment points to femur and tibia, makes reconstruction of the PCL technically challenging. The main pitfall is the sharp angle between created during reconstruction surgery bony tunnels, which forms so called "killer turn". The authors present a technique of the remnant-preserving PCL arthroscopic reconstruction that simplifies the procedure and uses the method of reverse passage of the PCL graft to overcome the killer turn.


Topology of the membrane-bound alkane hydroxylase of Pseudomonas oleovorans.
The Pseudomonas oleovorans alkane hydroxylase is an integral cytoplasmic membrane protein that is expressed and active in both Escherichia coli and P. oleovorans. Its primary sequence contains eight hydrophobic stretches that could span the membrane as alpha-helices. The topology of alkane hydroxylase was studied in E. coli using protein fusions linking different amino-terminal fragments of the alkane hydroxylase (AlkB) to alkaline phosphatase (PhoA) and to beta-galactosidase (LacZ). Four AlkB-PhoA fusions were constructed using transposon TnphoA. Site-directed mutagenesis was used to create PstI sites at 12 positions in AlkB. These sites were used to create AlkB-PhoA and AlkB-LacZ fusions. With respect to alkaline phosphatase and beta-galactosidase activity each set of AlkB-PhoA and AlkB-LacZ fusions revealed the expected complementary activities. At three positions, PhoA fusions were highly active, whereas the corresponding LacZ fusions were the least active. At all other positions the PhoA fusions were almost completely inactive, but the corresponding LacZ fusions were highly active. These data predict a model for alkane hydroxylase containing six transmembrane segments. In this model the amino terminus, two hydrophilic loops, and a large carboxyl-terminal domain are located in the cytoplasm. Only three very short loops near amino acid positions 52, 112, and 251 are exposed to the periplasm.


Responding to "Don't Say Gay" Laws in the US: Research Priorities and Considerations for Health Equity.
Despite increased legal rights for lesbian, gay, bisexual, transgender, and queer-identifying (LGBTQ +) people in the USA over the past 30 years, there has been an increasing number of anti-LGBTQ + laws proposed and passed at the state level. One of the most notorious laws, Florida's HB 1557, also known as the "Don't Say Gay" law, garnered substantial national attention for prohibiting discussions of sexual orientation or gender identity in public school classrooms. Other states quickly proposed similar laws, but little scholarship exists on the potential impacts of these laws. We explore the potential health equity ramifications of laws like Florida's HB 1557, focusing on the individual, interpersonal, and broader policy and practice implications. Examining these policies through the lens of political determinants of health, we identify theoretical and methodological approaches needed to address recent "Don't Say Gay" policies. Theoretical approaches emphasizing power, intersectionality, and the role of politics in health should guide research examining the impacts of recent anti-LGBTQ + policies. Laws like Florida's HB 1557 emphasize the need for methodological approaches that emphasize collaborative engagement between researchers and community members, and future research may be needed to understand how stressors created by law and policy can have individual and interpersonal consequences. Public health researchers have a role to play in reversing policies that negatively affect LGBTQ + individuals and undermine health equity. Research combating harmful policies may require theoretical approaches attentive to power differences and methodological approaches that squarely focus on disrupting power imbalances.


A randomized, double-blind, placebo-controlled, short-term monotherapy study of doravirine in treatment-naive HIV-infected individuals.
To assess the antiviral activity, pharmacokinetics, and safety of doravirine in nonnucleoside reverse transcriptase inhibitor-naïve, HIV-infected men. Double-blind, randomized, two-panel, dose-escalation study. In two sequential panels, 18 individuals received doravirine [25 mg (Panel A) or 200 mg (Panel B)] or matching placebo once daily for 7 days. Plasma samples were collected daily for measurement of HIV-1 RNA levels and doravirine pharmacokinetics. For the mean change from baseline in HIV RNA (log10 copies/ml) at 24 h after the day 7 dose, the mean difference (90% confidence interval) between doravirine and placebo was -1.37 (-1.60, -1.14) in the 25-mg group and -1.26 (-1.51, -1.02) in the 200-mg group. None of the participants had viral breakthrough. Increases in mean AUC0-24 h, Cmax, and C24 h were slightly less than dose-proportional, with median Tmax of 1.0-2.0 h. Steady state was achieved after 3-5 days of once-daily dosing. At steady state, accumulation ratios (day 7/day 1) for AUC0-24 h, Cmax, and C24 h were 1.2-1.6. The calculated effective t1/2 (10-16 h) was similar to that in HIV-uninfected individuals. Adverse events were limited in number, transient, and generally mild to moderate in intensity. One participant had a serious adverse event of elevated liver enzymes (judged probably not drug related) in concurrence with a newly acquired hepatitis C infection. Doravirine monotherapy demonstrated robust antiviral activity at both dose levels, without evidence of viral resistance, and was generally well tolerated. Doravirine pharmacokinetics in HIV-infected individuals were similar to those in uninfected individuals receiving similar doses in prior studies.


Factors associated with the overlap of frailty and nutrition in institutionalized older adults: A multicenter study.
This multicenter cross-sectional study aimed to investigate the factors associated with the overlap of frailty and nutrition in older adults residing in nursing homes. Older adults (n = 344, mean age ± SD = 77.7 ± 9.1) were selected from 17 nursing homes, 10 in Piracicaba and seven in João Pessoa, Brazil. The modified Fried's questionnaire and the Mini Nutrition Assessment Short-Form (MNA-SF) were used for screening frailty phenotypes and nutritional status, respectively. These variables overlapped into (1) frail and malnourished; (2) nonfrail and malnourished or frail and nourished; and (3) nonfrail and nourished. The independent variables were sex, age, mobility, number of medications, dependence for activities of daily living (ADL), maximal grip strength, muscle mass, body fat and body mass index (BMI). Spearman's correlation and Poisson regression were used to analyze the data (α = 0.05). The Spearman's test demonstrated a moderate correlation (r2=-0.441) between frailty (mean ± SD = 2.8 ± 1.7) and nutritional status (mean ± SD = 10.5 ± 2.9). Regarding the overlap of frailty and nutrition, 139 participants (40.5 %) were frail and malnourished, 121 (35.3 %) were nonfrail and malnourished or frail and nourished, and 83 (24.2 %) were nonfrail and nourished. The Poisson regression showed association of the coexistence of frailty and malnutrition with age (PR = 1.009, 95 % CI = 1.003-1.016), number of medications (PR = 1.016, 95 % CI = 1.006-1.027), dependence for ADL (PR = 1.061, 95 % CI = 1.016-1.108), maximal grip strength (PR = 0.992, 95 % CI = 0.986-0.998) and BMI (PR = 0.997, 95 % CI = 0.996-0.998). The findings showed that frailty and malnutrition were prevalent in institutionalized older adults and associated with advanced age, greater use of medicines, dependence for ADL, lower maximal grip strength and lower BMI.


Unbiased libraries in protein directed evolution.
Directed evolution is a powerful approach to study the molecular basis of protein evolution and to engineer proteins for a wide range of applications in synthetic organic chemistry and biotechnology. There are many methods based on random or focused mutagenesis to engineer successfully any protein trait. Focused approaches such as site-directed and saturation mutagenesis have become methods of choice for improving protein activity, selectivity, stability and many other traits because the screening step can be practically handled (bottleneck in directed evolution). Although novel mutagenesis methods based on CRISPR or solid-phase gene synthesis can eliminate bias when creating protein libraries, traditional PCR approaches, although imperfect, remain widely used due to their ease and low cost. One of the most common approaches in focused mutagenesis relies on NNK mutagenesis, however, the primer-based 22c-trick and small-intelligent methods have emerged as key tools for constructing less biased and unbiased libraries when all 20 canonical amino acids are needed for various reasons. In this minireview, we assess studies employing such methods for library creation and their areas of application. We also discuss the advantages and disadvantages of both methods and provide a perspective for creating smarter libraries.


Dermatomyofibromas Arising in Children: Report of Two New Cases and Review of the Literature.
Dermatomyofibroma is a rare, benign mesenchymal proliferation not commonly reported in children. Two patients with biopsy-proven dermatomyofibroma were identified (one female, one male) at our institution, both with rather atypical clinical presentations. The clinical and histopathologic findings and a review of the literature are presented here. Features of dermatomyofibroma should be recognized and differentiated from those of similar-appearing entities since the prognosis of this benign tumor is favorable after complete excision.


Do attending physicians, nurses, residents, and medical students agree on what constitutes medical student abuse?
Whether attending physicians, residents, nurses, and medical students agree on what constitutes medical student abuse, its severity, or influencing factors is unknown. We surveyed 237 internal medicine attending physicians, residents, medical students, and nurses at 13 medical schools after viewing five vignettes depicting potentially abusive behaviors. The majority of each group felt the belittlement, ethnic insensitivity, and sexual harassment scenarios represented abuse but that excluding a student from participating in a procedure did not. Only a majority of attending physicians considered the negative feedback scenario as abuse. Medical students rated abuse severity significantly lower than other groups in the belittlement scenario (p<.05). Respondents who felt abused as students were more likely to rate behaviors as abusive (p<.05). The groups generally agree on what constitutes abuse, but attending physicians and those abused as students may perceive more behaviors as abusive.


Passive smoking in childhood--tobacco smoke.
Prevalence of cigarette smoking varies widely in different countries, ranging, at the age of 13, from 2% to 5% (Sweden, United States) to more than 30% (Australia, Uruguay). Even if the prevalence of smokers among male adolescents is decreasing in western countries, it is increasing among girls and, in developing countries, male adolescent smokers still reach 40% (and up to 70%-80%). The determination of saliva cotinine levels, a product of nicotine metabolism, is an useful indicator of exposure to passive or active smoking. In a study of 210 children aged 9-13 years, we found detectable levels of saliva cotinine in 13% of children who lived in nonsmoking families and denied being regular smokers and in only 60% of children living in families with heavy cigarette consumption. This wide variations of a marker of smoking exposure may explain the differences in reported consequences of tobacco smoke in investigations conducted in different places and on different target populations. We later studied the effects of environmental tobacco smoke (ETS) in a sample of 166 nine-year-old children. The relationship between parental smoking and degree of bronchial responsiveness in males was significant. Also, prick skin test reactivity to allergens was significantly increased in children of smoking parents. Many studies concerning the effects of ETS exposure seem to demonstrate that not only the lungs and not only children are affected even if the difficulty of the epidemiology in such a field must suggest caution in interpreting the results.(ABSTRACT TRUNCATED AT 250 WORDS)


[11C]flumazenil binding is increased in a dose-dependent manner with tiagabine-induced elevations in GABA levels.
Evidence indicates that synchronization of cortical activity at gamma-band frequencies, mediated through GABA-A receptors, is important for perceptual/cognitive processes. To study GABA signaling in vivo, we recently used a novel positron emission tomography (PET) paradigm measuring the change in binding of the benzodiazepine (BDZ) site radiotracer [(11)C]flumazenil associated with increases in extracellular GABA induced via GABA membrane transporter (GAT1) blockade with tiagabine. GAT1 blockade resulted in significant increases in [(11)C]flumazenil binding potential (BPND) over baseline in the major functional domains of the cortex, consistent with preclinical studies showing that increased GABA levels enhance the affinity of GABA-A receptors for BDZ ligands. In the current study we sought to replicate our previous results and to further validate this approach by demonstrating that the magnitude of increase in [(11)C]flumazenil binding observed with PET is directly correlated with tiagabine dose. [(11)C]flumazenil distribution volume (VT) was measured in 18 healthy volunteers before and after GAT1 blockade with tiagabine. Two dose groups were studied (n = 9 per group; Group I: tiagabine 0.15 mg/kg; Group II: tiagabine 0.25 mg/kg). GAT1 blockade resulted in increases in mean (± SD) [(11)C]flumazenil VT in Group II in association cortices (6.8 ± 0.8 mL g-1 vs. 7.3 ± 0.4 mL g-1;p = 0.03), sensory cortices (6.7 ± 0.8 mL g-1 vs. 7.3 ± 0.5 mL g-1;p = 0.02) and limbic regions (5.2 ± 0.6 mL g-1 vs. 5.7 ± 0.3 mL g-1;p = 0.03). No change was observed at the low dose (Group I). Increased orbital frontal cortex binding of [(11)C]flumazenil in Group II correlated with the ability to entrain cortical networks (r = 0.67, p = 0.05) measured via EEG during a cognitive control task. These data provide a replication of our previous study demonstrating the ability to measure in vivo, with PET, acute shifts in extracellular GABA.


Termination of ouabain-induced ventricular tachycardia by flunarizine in conscious dogs.
The ability of flunarizine to terminate ouabain-induced ventricular tachycardia was investigated in conscious dogs. These arrhythmias result from triggered activity based on delayed afterdepolarizations. Sustained ventricular tachycardia was induced by ouabain (48 +/- 6 micrograms/kg) and pacing in 13 animals with surgically induced complete atrioventricular block. Flunarizine was administered i.v. when the arrhythmia had persisted for at least 20 min. Flunarizine induced an increase of the R-R interval from 300 +/- 30 to 410 +/- 50 ms (P less than or equal to 0.001) in all dogs. Flunarizine (2 mg/kg) terminated ventricular tachycardia with long R-R intervals (330 +/- 20 ms) in seven dogs. An additional dose of flunarizine (1 mg/kg) resulted in termination of the faster tachycardias (280 +/- 15 ms). The arrhythmias were not induced again by pacing for 27 +/- 18 min. Reinitiated tachycardias could be suppressed again by flunarizine. In conclusion, flunarizine (1) slows and terminates arrhythmias resulting from delayed afterdepolarizations, and (2) prevents their reinitiation.


Inhibition of γ/β Globin Gene Switching in CD 34+ Derived Erythroid Cells by BCL11A RNA Silencing.
The induction of fetal haemoglobin (Hb F), due to the sustained clinical effects, is one of the most promising methods for the treatment of β hemoglobinopathies, such as thalassemia major and sickle cell disease (SCD). Inhibition of γ-globin gene silencing, possibly is a suitable strategy to induce HbF expression in these patients. In this study, the possibility of increasing HbF in the CD34+ derived erythroid cells was investigated by BCL11A inhibition using specific small-interfering RNAs (siRNAs). Human peripheral blood-derived hematopoietic stem cells were isolated and differentiated to erythroid cells. Erythroid maturation was investigated using cell morphology parameters and flow cytometry analysis of CD235a expression On day 20, siRNA complementary to BCL11A was transfected to differentiating cells via electroporation. BCL11A expression was evaluated through real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) and enzyme linked immunosorbant assay (ELISA). β actin was used as the reference gene to confirm the relative expression level of BCL11A gene mRNA. 48 hours after transfection, BCL11A siRNA significantly reduced BCL11A mRNA levels and consequently led to 2.0 fold decrease in corresponding protein. On the 28th day, haemoglobin electrophoresis results showed that Hb F levels in transfected erythroid cells increased 3.3-fold when compared with non transfected cells. In this study we showed that BCL11A inhibition in erythroid cells could increase fetal hemoglobin, and this strategy can be the basis for designing a γ globin expressing cellular system to increase Hb F in patients with thalassemia and SCD.


Optical and imaging properties of a novel multi-segment spectacle lens designed to slow myopia progression.
High sampling density optical metrology combined with pupil- and image-plane numerical analyses were applied to evaluate a novel spectacle lens containing multiple small zones designed to slow myopia progression. High-resolution aberrometry (ClearWave, www.lumetrics.com) was used to sample wavefront slopes of a novel spectacle lens, Defocus Incorporated Multiple Segments (DIMS) (www.hoya.com), incorporating many small, positive-powered lenslets in its periphery. Using wavefront slope and error maps, custom MATLAB software ('Indiana Wavefront Analyzer') was used to compute image-plane point-spread functions (PSF), modulation transfer functions (MTF), simulated images and power distributions created by the dual-focus optic for different pupil sizes and target vergences. Outside of a central 10 mm zone containing single distance optical power, a hexagonal array of small 1 mm lenslets with nearest-neighbour separations of 0.5 mm were distributed over the lens periphery. Sagittal and curvature-based measures of optical power imperfectly captured the consistent +3.50 D add produced by the lenslets. Image plane simulations revealed multiple PSFs and poor image quality at the lenslet focal plane. Blur at the distance optic focal plane was consistent with a combination of diffraction blur from the distance optic and the approximately +3.50 D of defocus from the 1 mm diameter near optic zones. Converging the defocused beams generated by the multiple small (1 mm diameter) lenslets to a blurred image at the distance focal plane produced a blur magnitude determined by the small lenslet diameter and not the overall pupil diameter. The distance optic located in between the near-add lenslets determines the limits of the optical quality achievable by the lens. When compared to the optics of a traditional concentric-zone dual-focus contact lens, the optics of the DIMS lens generates higher-contrast images at low spatial frequencies (<7 cycles per degree), but lower-contrast at high spatial frequencies.


Role of the malonyl-CoA synthetase ACSF3 in mitochondrial metabolism.
Malonyl-CoA is a central metabolite in fatty acid biochemistry. It is the rate-determining intermediate in fatty acid synthesis but is also an allosteric inhibitor of the rate-setting step in mitochondrial long-chain fatty acid oxidation. While these canonical cytoplasmic roles of malonyl-CoA have been well described, malonyl-CoA can also be generated within the mitochondrial matrix by an alternative pathway: the ATP-dependent ligation of malonate to Coenzyme A by the malonyl-CoA synthetase ACSF3. Malonate, a competitive inhibitor of succinate dehydrogenase of the TCA cycle, is a potent inhibitor of mitochondrial respiration. A major role for ACSF3 is to provide a metabolic pathway for the clearance of malonate by the generation of malonyl-CoA, which can then be decarboxylated to acetyl-CoA by malonyl-CoA decarboxylase. Additionally, ACSF3-derived malonyl-CoA can be used to malonylate lysine residues on proteins within the matrix of mitochondria, possibly adding another regulatory layer to post-translational control of mitochondrial metabolism. The discovery of ACSF3-mediated generation of malonyl-CoA defines a new mitochondrial metabolic pathway and raises new questions about how the metabolic fates of this multifunctional metabolite intersect with mitochondrial metabolism.