Prefrontal cortex involvement in preattentive auditory deviance detection: neuroimaging and electrophysiological evidence.
Previous electrophysiological and neuroimaging studies suggest that the mismatch negativity (MMN) is generated by a temporofrontal network subserving preattentive auditory change detection. In two experiments we employed event-related brain potentials (ERP) and event-related functional magnetic resonance imaging (fMRI) to examine neural and hemodynamic activity related to deviance processing, using three types of deviant tones (small, medium, and large) in both a pitch and a space condition. In the pitch condition, hemodynamic activity in the right superior temporal gyrus (STG) increased as a function of deviance. Comparisons between small and medium and between small and large deviants revealed right prefrontal activation in the inferior frontal gyrus (IFG; BA 44/45) and middle frontal gyrus (MFG; BA 46), whereas large relative to medium deviants led to left and right IFG (BA 44/45) activation. In the ERP experiment the amplitude of the early MMN (90-120 ms) increased as a function of deviance, by this paralleling the right STG activation in the fMRI experiment. A U-shaped relationship between MMN amplitude and the degree of deviance was observed in a late time window (140-170 ms) resembling the right IFG activation pattern. In a subsequent source analysis constrained by fMRI activation foci, early and late MMN activity could be modeled by dipoles placed in the STG and IFG, respectively. In the spatial condition no reliable hemodynamic activation could be observed. The MMN amplitude was substantially smaller than in the pitch condition for all three spatial deviants in the ERP experiment. In contrast to the pitch condition it increased as a function of deviance in the early and in the late time window. We argue that the right IFG mediates auditory deviance detection in case of low discriminability between a sensory memory trace and auditory input. This prefrontal mechanism might be part of top-down modulation of the deviance detection system in the STG.


Neuromuscular paralysis for newborn infants receiving mechanical ventilation.
Ventilated newborn infants breathing in asynchrony with the ventilator are at risk for complications during mechanical ventilation, such as pneumothorax or intraventricular hemorrhage, and are exposed to more severe barotrauma, which consequently could impair their clinical outcome. Neuromuscular paralysis, which eliminates spontaneous breathing efforts of the infant, has potential advantages in this respect. However, a number of complications have been reported with muscle relaxation in infants, so that concerns exist regarding the safety of prolonged neuromuscular paralysis in newborn infants. To determine whether routine neuromuscular paralysis of newborn infants receiving mechanical ventilation compared with no routine paralysis results in clinically important benefits or harms. MEDLINE (from 1966 to May 2000) and EMBASE (from 1988 to May 2000) were searched, as well as The Cochrane Controlled Trials Register (issue 2, 2000). References of review articles were hand searched. Language restriction was not imposed. All trials using random or quasi-random patient allocation, in which the routine use of neuromuscular blocking agents during mechanical ventilation was compared to no paralysis or selective paralysis in newborn infants. Methodological quality was assessed blindly and independently by the two authors. Data were abstracted using standard methods of the Cochrane Collaboration and its Neonatal Review Group, with independent evaluation of trial quality, and abstraction and synthesis of data by both authors. Treatment effect was analysed using relative risk, risk difference and weighted mean difference. Ten possibly eligible trials were identified, of which five were included in the review. All the included trials studied preterm infants ventilated for respiratory distress syndrome, and used pancuronium as the neuromuscular blocking agent. In the analysis of the results of all trials, no difference was found in mortality, air leak or chronic lung disease, but there was a significant reduction in intraventricular hemorrhage and a trend towards less severe intraventricular hemorrhages. In the subgroup analysis of trials studying a selected population of ventilated infants with evidence of asynchronous respiratory efforts, a significant reduction in intraventricular hemorrhage (any grade and severe IVH) was found, and a trend towards less air leak. In the subgroup analysis of trials studying an unselected population of ventilated infants, no differences were found for any of the outcomes. For ventilated preterm infants with evidence of asynchronous respiratory efforts, neuromuscular paralysis with pancuronium seems to have a favourable effect on intraventricular hemorrhage and possibly on air leak. Uncertainty remains, however, regarding the long term pulmonary and neurologic effects, and regarding the safety of prolonged use of pancuronium in ventilated newborn infants. There is no evidence from randomized trials on the effects of neuromuscular blocking agents other than pancuronium. Therefore, the routine use of pancuronium or any other neuromuscular blocking agent in ventilated newborn infants cannot be recommended based on current evidence.


Proline conformations in linear peptides. Structure determination of the methyl ester of N-benzyloxycarbonyl-L-prolyl-D-alanine (N-Z-L-Pro-D-Ala-OMe).
N-Z-L-Pro-D-Ala-OMe, C17H22N2O5, Mr = 334.38, crystallizes in the orthorhombic space group P2(1)2(1)2(1), with the cell dimensions a = 5.005 (5), b = 17.690 (9) and c = 18.70 (1) A3, V = 1656.3 A3, Z = 4, Dx = 1.341 g cm-3, Cu K alpha, lambda = 1.54184 A, mu = 7.830 cm-1, F(000) = 712, T = 198 K, final R (on F) = 0.036 for 1575 observed reflections with I greater than or equal to 3 sigma(I). The pyrrolidine ring takes on the C2-C gamma-endo conformation. The urethane bond is in the cis conformation [omega 0 = 6.0 (3) degrees] while the peptide bond is in the trans conformation [omega 1 = 170.8 (2) degrees]; phi 1/psi 1 values are -88.0 (3) degrees and 151.3 (2) degrees. Intermolecular hydrogen bonding occurs between the C-terminus and the symmetry-related amide. Systematic examination of the pyrrolidine ring in linear peptides reveals no correlation exists between the cis-trans orientation of the proline and the conformation of the pyrrolidine ring.


Predation risk constrains herbivores' adaptive capacity to warming.
Global warming compels larger endothermic animals to adapt either physiologically or behaviourally to avoid thermal stress, especially in tropical ecosystems. Their adaptive responses may however be compromised by other constraints, such as predation risk or starvation. Using an exceptional camera-trap dataset spanning 32 protected areas across southern Africa, we find that intermediate-sized herbivores (100-550 kg) switch activity to hotter times of the day when exposed to predation by lions. These herbivores face a tight window for foraging activity being exposed to nocturnal predation and to heat during the day, suggesting a trade-off between predation risk and thermoregulation mediated by body size. These findings stress the importance of incorporating trophic interactions into climate change predictions.


Disparate gain and loss of parasitic abilities among nematode lineages.
Plant parasitism has arisen time and again in multiple phyla, including bacteria, fungi, insects and nematodes. In most of these organismal groups, the overwhelming diversity hampers a robust reconstruction of the origins and diversification patterns of this trophic lifestyle. Being a moderately diversified phylum with ≈ 4,100 plant parasites (15% of total biodiversity) subdivided over four independent lineages, nematodes constitute a major organismal group for which the genesis of plant parasitism could be mapped. Since substantial crop losses worldwide have been attributed to less than 1% of these plant parasites, research efforts are severely biased towards this minority. With the first molecular characterisation of numerous basal and supposedly harmless plant parasites as well as their non-parasitic relatives, we were able to generate a comprehensive molecular framework that allows for the reconstruction of trophic diversification for a complete phylum. In each lineage plant parasites reside in a single taxonomic grouping (family or order), and by taking the coverage of the next lower taxonomic level as a measure for representation, 50, 67, 100 and 85% of the known diversity was included. We revealed distinct gain and loss patterns with regard to plant parasitism per se as well as host exploitation strategies between these lineages. Our map of parasitic nematode biodiversity also revealed an unanticipated time reversal in which the two most ancient lineages showed the lowest level of ecological diversification and vice versa.


A comparative study of psychotic and affective symptoms in Rwandan and Kenyan students.
War and conflict are known to adversely affect mental health, although their effects on risk symptoms for psychosis development in youth in various parts of the world are unclear. The Rwandan genocide of 1994 and Civil War had widespread effects on the population. Despite this, there has been no significant research on psychosis risk in Rwanda. Our goal in the present study was to investigate the potential effects of genocide and war in two ways: by comparing Rwandan youth born before and after the genocide; and by comparing Rwandan and Kenyan adolescents of similar age. A total of 2255 Rwandan students and 2800 Kenyan students were administered the Washington Early Recognition Center Affectivity and Psychosis (WERCAP) Screen. Prevalence, frequency and functional impairment related to affective and psychosis-risk symptoms were compared across groups using univariate and multivariate statistics. Rwandan students born before the end of the genocide and war in 1994 experienced higher psychotic and affective symptom load (p's < 0.001) with more functional impairment compared with younger Rwandans. 5.35% of older Rwandan students met threshold for clinical high-risk of psychosis by the WERCAP Screen compared with 3.19% of younger Rwandans (χ 2 = 5.36; p = 0.02). Symptom severity comparisons showed significant (p < 0.001) group effects between Rwandan and Kenyan secondary school students on affective and psychotic symptom domains with Rwandans having higher symptom burden compared with Kenyans. Rwandan female students also had higher rates of psychotic symptoms compared with their male counterparts - a unique finding not observed in the Kenyan sample. These results suggest extreme conflict and disruption to country from genocide and war can influence the presence and severity of psychopathology in youth decades after initial traumatic events.


Artificial iron hydrogenase made by covalent grafting of Knölker's complex into xylanase: Application in asymmetric hydrogenation of an aryl ketone in water.
We report a new artificial hydrogenase made by covalent anchoring of the iron Knölker's complex to a xylanase S212C variant. This artificial metalloenzyme was found to be able to catalyze efficiently the transfer hydrogenation of the benchmark substrate trifluoroacetophenone by sodium formate in water, yielding the corresponding secondary alcohol as a racemic. The reaction proceeded more than threefold faster with the XlnS212CK biohybrid than with the Knölker's complex alone. In addition, efficient conversion of trifluoroacetophenone to its corresponding alcohol was reached within 60 H with XlnS212CK, whereas a ≈2.5-fold lower conversion was observed with Knölker's complex alone as catalyst. Moreover, the data were rationalized with a computational strategy suggesting the key factors of the selectivity. These results suggested that the Knölker's complex was most likely flexible and could experience free rotational reorientation within the active-site pocket of Xln A, allowing it to access the subsite pocket populated by trifluoroacetophenone.


Phase I study of alpelisib (BYL719), an α-specific PI3K inhibitor, in Japanese patients with advanced solid tumors.
This phase I study aimed to determine tolerability and preliminary efficacy of single-agent alpelisib (BYL719) in Japanese patients with advanced solid malignancies. The primary objective of the study was to estimate the maximum tolerated dose (MTD) and/or recommended phase II dose (RP2D) of oral alpelisib in patients with advanced solid tumors who had progressed despite standard therapy. The expansion part included patients with PIK3CA mutation/amplification; safety, preliminary efficacy, pharmacokinetic (PK)/pharmacodynamic profile, and food effect on the PK profile of alpelisib at the MTD/RP2D were determined. Oral alpelisib was given as a single agent on a continuous 28-day treatment cycle once daily. Overall, 33 patients received alpelisib. Dose-limiting toxicities were observed in 2 patients in the escalation part (at 400 mg/day) and 1 patient in the expansion part (at 350 mg/day). The RP2D of alpelisib was determined as 350 mg/day based on overall safety profile in the dose escalation part and previous data from a Western population; the MTD was not determined. The most common all-grade treatment-suspected adverse events were hyperglycemia and maculopapular rash (48.5% each) and diarrhea (45.5%). The PK of alpelisib in the Japanese population was similar to that reported in the Western population. The overall response rate, disease control rate, and median progression-free survival at 350 mg/day were 3%, 57.6%, and 3.4 months, respectively. Alpelisib as single agent showed a favorable safety profile and encouraging preliminary efficacy in Japanese patients with advanced solid tumors.


Semi-Local Scaling Exponent Estimation With Box-Penalty Constraints and Total-Variation Regularization.
We here establish and exploit the result that 2D isotropic self-similar fields beget quasi-decorrelated wavelet coefficients and that the resulting localised log sample second moment statistic is asymptotically normal. This leads to the development of a semi-local scaling exponent estimation framework with optimally modified weights. Furthermore, recent interest in penalty methods for least square problems and generalized Lasso for scaling exponent estimation inspires the simultaneous incorporation of both bounding box constraints and total variation smoothing into an iteratively reweighted least-square estimator framework. Numerical results on fractional Brownian fields with global and piecewise constant, semi-local Hurst parameters illustrate the benefits of the new estimators.


Diagnostic characteristics of late-night salivary cortisol using liquid chromatography-tandem mass spectrometry.
The objective of the study was to describe the diagnostic performance of a commercially available late-night salivary cortisol (NSC) assay using liquid chromatography tandem mass spectrometry. We retrospectively identified 90 patients who had one or more NSC determinations: 52 patients in whom Cushing syndrome (CS) was excluded or could not be confirmed [group 1 (G1)] and 38 patients in whom CS was confirmed [group 2 (G2)]. Eighteen healthy volunteers served as controls. Baseline demographics in all groups were similar with regards to age, ethnicity, gender, and body mass index. NSC levels [median (range)] were higher in G2, 381 (64-13,500) ng/dl [10.51 (1.77-372.46) nmol/liter], compared with controls, 19.3 (2.1-416) ng/dl [0.53 (0.06-11.48) nmol/liter], and G1, 26 (4-176) ng/dl [0.72 (0.11-4.86) nmol/liter, P < 0.001]. The highest combined sensitivity (92%) and specificity (92%) was achieved at a cut point of 107 ng/dl (2.95 nmol/liter). Two or more NSCs were done in 32 of 52 G1 and 31 of 38 G2 patients. In G1 eight of 32 (25%) had at least one elevated [>100 ng/dl (2.76 nmol/liter)] NSC including two in whom both NSCs were elevated. In contrast, four of 31 (13%) in G2 had at least one normal NSC including one with four of five normal NSC values. None of the patients with CS had a NSC less than 60 ng/dl (<1.66 nmol/liter). Comparing G1 and G2, obtaining more than one saliva sample did not improve the diagnostic accuracy of NSC measurement (P = 0.64). The liquid chromatography tandem mass spectrometry assay to measure NSC is a simple and reliable test to screen patients suspected to have CS. Clinicians should be aware of appropriate cutoff values for proper interpretation of NSC and use additional tests when necessary.


The 'fixed cost effect' on practice management.
To obtain a better understanding of the behavior of "non-professional" costs in a medical practice, the authors analyzed the expenses of a 19-doctor practice. The analysis revealed that 80 percent of these expenses were fixed costs. Fixed costs, as opposed to variable costs, remain static in total but vary on a per unit basis as volume changes. Organizations with high fixed cost must maximize capacity to achieve profitability. Thus, the relationship among volume, capacity, cost and profit must be understood by medical practices negotiating rates for service units.


Nodding Duck Structure Multi-track Directional Freestanding Triboelectric Nanogenerator toward Low-Frequency Ocean Wave Energy Harvesting.
Development of ocean energy conversion technique is a strategic requirement to optimize the energy structure and expand the "blue economic" space. Triboelectric nanogenerator (TENG) provides a potential approach for efficiently capturing wave energy with its unique advantages. Herein, a nodding duck structure multi-track freestanding triboelectric-layer nanogenerator (NDM-FTENG) is developed for ocean wave energy harvesting at a low-frequency range. Configuration parameters including track numbers, connection approach, oscillation frequency, and swing amplitude on electrical output performances of NDM-FTENG are systematically investigated and optimized; the maximum instantaneous power density of 4 W/m3 is obtained by one NDM-FTENG block with 320 LEDs lighted up simultaneously. Overall, NDM-FTENG is proved to be an efficient device for driving small electronics with excellent stability and durability in a real water wave environment, and the power potential can be further magnified by combining more NDM-FTENG devices in parallel to form a network toward large-scale blue energy harvesting.


Methylenetetrahydrofolate reductase (MTHFR) C677T polymorphism is associated with osteoporotic vertebral fractures, but is a weak predictor of BMD.
Osteoporosis is a common disease with a strong genetic component. Linkage studies have suggested linkage between BMD and loci on chromosome 1. The MTHFR gene is located on chromosome 1. MTHFR catalyzes the conversion of 5,10-methylenetetrahydrofolate to 5-methylenetetrahydrofolate, which is used for homocysteine methylation to methionine. The rare genotype (TT) of the C677T polymorphism has previously been demonstrated to be associated with increased plasma homocysteine levels in individuals with inadequate plasma folate levels. Recently, the TT genotype has been found to be associated with reduced bone mass. We therefore examined if the C677T polymorphism in the MTHFR gene is associated with changes in bone mass and risk of osteoporotic fractures in 388 osteoporotic patients and 336 normal individuals. The distributions of the genotypes CC, CT and TT in women with osteoporotic vertebral fractures and normal controls were 43.5%, 42.2% and 14.3% and 52.0%, 42.0% and 8.0%, respectively, chi2 = 5.62, P = 0.06. Since studies of the functionality of this polymorphism have revealed that only the TT genotype is associated with biochemical changes, we also compared the prevalence of the TT genotype versus the CT- and CC genotypes in patients and controls and found that the TT genotype is significantly more common in women with vertebral fractures (14.3%) compared with normal controls (8.0%), chi2 = 4.31, P < 0.05. Logistic regression analysis demonstrated that vertebral fractures were significantly associated with BMD (lumbar spine) and height but only marginally with the MTHFR genotype (P = 0.06). Multiple linear regression analysis revealed that weight, age and the MTHFR polymorphism were predictors of lumbar spine BMD in women. However, age- and gender-corrected BMD of the lumbar spine and the hip was not significantly different between MTHFR genotypes. Furthermore, individuals with the TT genotype did not have BMD significantly lower than the combined group of individuals with the CT- or CC genotypes. In conclusion, we have demonstrated that the rare TT genotype of the C677T polymorphism in the MTHFR gene is associated with increased risk of osteoporotic fractures in women and a weak predictor of lumbar spine BMD.


[Complications and mortality of extended gastrectomy for gastric cancer].
Extended gastrectomy allows a complete tumor excision in patients with advanced gastric cancer. To compare the surgical results of extended (ER) and non-extended gastrectomy (NER) among patients with gastric cancer, and determine factors associated with complications and mortality. Review of medical records of patients with gastric cancer who underwent complete resection between 2002 and 2008 in an oncological hospital. Demographics, patient-related and therapeutic features were compared between groups, and independent factors were established with multivariate analysis. Seventy four patients, (44 men, median age 62 years) underwent an ER and 103 patients, (56 men, median age 61 years) a NER. Specifically, ER included splenectomy alone in 27 patients, splenectomy associated with other procedure in 24, partial esophagectomy in 18, distal pancreatectomy in 13, hemicolectomy in 8, total esophagectomy in 7, partial hepatectomy in 4, and adrenalectomy in 1. Postoperative complications were observed in 19 patients treated with an ER (26%) and in 11 patients treated with a NER (11%), p < 0.05. Serious complications were higher in patients who underwent an ER compared with NER (6 patients (8%) vs. 4 (4%), respectively) p < 0.05. In the same way, mortality was higher in patients treated with ER when it was compared with NER (4 patients (5%) vs two (2%), respectively), p < 0.05. ER and serum albumin levels were independent factors associated to a higher risk of mortality and rate of complications. ER was associated with a higher rate of general and severe complications, and mortality.


Salmonella spp. contamination in commercial layer hen farms using different types of samples and detection methods.
The performance of detection methods (culture methods and polymerase chain reaction assay) and plating media used in the same type of samples were determined as well as the specificity of PCR primers to detected Salmonella spp. contamination in layer hen farms. Also, the association of farm characteristics with Salmonella presence was evaluated. Environmental samples (feces, feed, drinking water, air, boot-swabs) and eggs were taken from 40 layer hen houses. Salmonella spp. was most detected in boot-swabs taken around the houses (30% and 35% by isolation and PCR, respectively) follow by fecal samples (15.2% and 13.6% by isolation and PCR, respectively). Eggs, drinking water, and air samples were negative for Salmonella detection. Salmonella Schwarzengrund and S. Enteritidis were the most isolated serotypes. For plating media, relative specificity was 1, and the relative sensitivity was greater for EF-18 agar than XLDT agar in feed and fecal samples. However, relative sensitivity was greater in XLDT agar than EF-18 agar for boot-swab samples. Agreement was between fair to good depending on the sample, and it was good between isolation and PCR (feces and boot-swabs), without agreement for feed samples. Salmonella spp. PCR was positive for all strains, while S. Typhimurium PCR was negative. Salmonella Enteritidis PCR used was not specific. Based in the multiple logistic regression analyses, categorization by counties was significant for Salmonella spp. presence (P-value = 0.010). This study shows the importance of considering different types of samples, plating media and detection methods during a Salmonella spp. monitoring study. In addition, it is important to incorporate the sampling of floors around the layer hen houses to learn if biosecurity measures should be strengthened to minimize the entry and spread of Salmonella in the houses. Also, the performance of some PCR methods and S. Enteritidis PCR should be improved, and biosecurity measures in hen farms must be reinforced in the region of more concentrated layer hen houses to reduce the probability of Salmonella spp. presence.


Pharmacokinetics and pharmacodynamics of glycopyrrolate following a continuous-rate infusion in the horse.
Glycopyrrolate (GLY) is an antimuscarinic agent that is used in humans and domestic animals primarily to reduce respiratory tract secretions during anesthesia and to reverse intra-operative bradycardia. Although GLY is used routinely in veterinary patients, there is limited information regarding its pharmacokinetic (PK) and pharmacodynamic (PD) properties in domestic animals, and an improved understanding of the plasma concentration-effect relationship in racehorses is warranted. To accomplish this, we characterize the pharmacokinetic-pharmacodynamic (PK-PD) actions of GLY during and after a 2-h constant-rate intravenous infusion (4 μg/kg/h) and evaluate potential PK-PD models for cardiac stimulation in adult horses. Measurements of plasma GLY concentrations, heart and respiration rates, and frequency of bowel movements were performed in six Thoroughbred horses. The time course for GLY disposition in plasma followed a tri-exponential equation characterized by rapid disappearance of GLY from blood followed by a prolonged terminal phase. Physiological monitoring revealed significant (P < 0.01) increases in heart (>70 bpm) and respiratory rates accompanied by a marked and sustained delay in the frequency of bowel movements (1.1 ± 0.2 h [saline group] vs. 6.0 ± 2.0 h [GLY group]). Two of six horses showed signs of colic during the 8-h observation period after the end of the GLY infusion, but were treated and recovered without further complications. The relationship between plasma GLY concentration and heart rate exhibited counterclockwise hysteresis that was adequately described using an effect compartment.


Contribution of cytogenetics to the management of poorly differentiated sarcomas.
Sarcomas constitute a heterogeneous group of rare tumors that in recent years have been shown by cytogenetic analysis to have a remarkably high incidence of specific and primary alterations. These genetic alterations not only have guided molecular studies in establishing the underlying genes involved, thereby yielding important pathogenetic information, but have also provided clinicians with a valuable tool to add to their diagnostic armamentarium. The addition of molecular cytogenetic (fluorescence in situ hybridization [FISH]) and molecular approaches (reverse transcriptase-polymerase chain reaction [RT-PCR]) has further enhanced the sensitivity and accuracy of detecting nonrandom chromosomal imbalances and/or structural rearrangements in sarcomas, including assessment in formalin-fixed, paraffin-embedded tissues. Poorly differentiated sarcomas represent a significant challenge to the pathologist as these neoplasms lack an identifiable hematoxylin and eosin-stained phenotype and often have lost diagnostic immunohistochemical or ultrastructural features as well. In contrast, primary cytogenetic changes and associated molecular events such as the 11;22 translocation in Ewing sarcoma are retained as a given tumor metastasizes or becomes less differentiated, as their presence appears to be vital for sustaining neoplastic transformation. Consequently, demonstration of characteristic, tumor-specific chromosomal aberrations is especially useful in the management of poorly differentiated sarcomas.


Extant mammal body masses suggest punctuated equilibrium.
Is gradual microevolutionary change within species simultaneously the source of macroevolutionary differentiation between species? Since its first publication, Darwin's original idea that phenotypic differences between species develop gradually over time, as the accumulation of small selection-induced changes in successive generations has been challenged by palaeontologists claiming that, instead, new species quickly acquire their phenotypes to remain virtually unchanged until going extinct again. This controversy, widely known as the 'punctuated equilibrium' debate, remained unresolved, largely owing to the difficulty of distinguishing biological species from fossil remains. We analysed body masses of 2143 existing mammal species on a phylogeny comprising 4510 (i.e. nearly all) extant species to estimate rates of gradual (anagenetic) and speciational (cladogenetic) evolution. Our Bayesian estimates from mammals as well as separate sub-clades such as primates and carnivores suggest that gradual evolution is responsible for only a small part of body size variation between mammal species.


Detecting copy number variations from array CGH data based on a conditional random field model.
Array comparative genomic hybridization (aCGH) allows identification of copy number alterations across genomes. The key computational challenge in analyzing copy number variations (CNVs) using aCGH data or other similar data generated by a variety of array technologies is the detection of segment boundaries of copy number changes and inference of the copy number state for each segment. We have developed a novel statistical model based on the framework of conditional random fields (CRFs) that can effectively combine data smoothing, segmentation and copy number state decoding into one unified framework. Our approach (termed CRF-CNV) provides great flexibilities in defining meaningful feature functions. Therefore, it can effectively integrate local spatial information of arbitrary sizes into the model. For model parameter estimations, we have adopted the conjugate gradient (CG) method for likelihood optimization and developed efficient forward/backward algorithms within the CG framework. The method is evaluated using real data with known copy numbers as well as simulated data with realistic assumptions, and compared with two popular publicly available programs. Experimental results have demonstrated that CRF-CNV outperforms a Bayesian Hidden Markov Model-based approach on both datasets in terms of copy number assignments. Comparing to a non-parametric approach, CRF-CNV has achieved much greater precision while maintaining the same level of recall on the real data, and their performance on the simulated data is comparable.


The monocytosis during human leptospirosis is associated with modest immune cell activation states.
Leptospirosis is a life-threatening zoonotic disease and it has been hypothesized that the innate immune system fails to control the infection through ill-characterized mechanisms. The aim of this observational study was to better evaluate the activation processes of monocytes at the early stage of the disease. Blood samples were taken from healthy donors (n = 37) and patients hospitalized for either non-severe (n = 25) or severe (n = 32) leptospirosis. Monocyte cell counts and phenotypes were assessed by flow cytometry. We analysed the expression of several cell activation markers: CD14, CD16, HLA-DR, CD69, TLR2, TLR4, CD11b and CD11c. Although monocyte values at admittance were not significantly different from controls, patients experienced significant monocytosis at 1.33 × 109/L (p < 0.0001 compared to controls: 0.56 × 109/L) during their hospital stay. This monocytosis observed during hospital stay was correlated to several surrogate markers of organ injury. Non-classical (CD14-CD16+) and intermediate (CD14+CD16+) monocyte subsets increased compared to controls (p < 0.05). Accordingly, classical monocyte subset (CD14+CD16-) showed decreased percentages (p < 0.0001). Levels of several cell surface activation molecules were decreased: HLA-DR involved in MHC class II antigen presentation, integrins CD11b and CD11c implicated in phagocytosis and cell recruitment (p < 0.0001). None of these parameters had a prognostic value. Results from this study showed that during acute human leptospirosis, patients experienced monocytosis with a switch toward an inflammation-related phenotype contrasted by low expression levels of markers implicated in monocyte function.


A mouse ocular explant model that enables the study of living optic nerve head events after acute and chronic intraocular pressure elevation: Focusing on retinal ganglion cell axons and mitochondria.
We developed an explant model of the mouse eye and optic nerve that facilitates the study of retinal ganglion cell axons and mitochondria in the living optic nerve head (ONH) in an ex vivo environment. Two transgenic mouse strains were used, one expressing yellow fluorescent protein in selected axons and a second strain expressing cyan fluorescent protein in all mitochondria. We viewed an explanted mouse eye and optic nerve by laser scanning microscopy at and behind the ONH, the site of glaucoma injury. Explants from previously untreated mice were studied with the intraocular pressure (IOP) set artificially at normal or elevated levels for several hours. Explants were also studied from eyes that had undergone chronic IOP elevation from 14 h to 6 weeks prior to ex vivo study. Image analysis in static images and video of individual mitochondria or axonal structure determined effects of acute and chronic IOP elevation. At normal IOP, fluorescent axonal structure was stable for up to 3 h under ex vivo conditions. After chronic IOP elevation, axonal integrity index values indicated fragmentation of axon structure in the ONH. In mice with fluorescent mitochondria, the normal density decreased with distance behind the ONH by 45% (p = 0.002, t-test). Density increased with prior chronic IOP elevation to 21,300 ± 4176 mitochondria/mm2 compared to control 16,110 ± 3159 mitochondria/mm2 (p = 0.025, t-test), but did not increase significantly after 4 h, acute IOP elevation (1.5% decrease in density, p = 0.83, t-test). Mean normal mitochondrial length of 2.3 ± 1.4 μm became 13% smaller after 4 h of IOP elevation ex vivo compared to baseline (p = 0.015, t-test, N-10). Normal mitochondrial speed of movement was significantly slower in the anterograde direction (towards the brain) than retrograde, but there were more mitochondria in motion and traveling longer lengths in anterograde direction. The percent of mitochondria in motion decreased by >50% with acute IOP increase to 30 mm Hg after 60 min. A new ocular explant model implemented with eyes from transgenic mice with fluorescent cellular components provided real time measurement of the early events in experimental glaucoma and quantitative outcomes for neuroprotection therapy experiments.


Kinetics of deposition and clearance of inhaled mineral dusts during chronic exposure.
New inhalation studies have been carried out with rats exposed to UICC (Union International Contre le Cancer) amosite asbestos, with the main aim of further elucidating the factors the influence the accumulation of dust in the lung during prolonged chronic exposure. The results show that, for exposure times beyond a few weeks, the lung burden rises linearly and does not level off as predicted by simple models based on ideas taken from the 1966 report of the Task Group on Lung Dynamics. Furthermore, the lung burden is found to scale directly in proportion to the exposure concentration in a way that seems to contradict the overload hypothesis stated earlier. Nevertheless, the general pattern exhibited by our results for asbestos is markedly similar to that found elsewhere for rats inhaling diesel fume, leading to the suggestion that it is general (and not specific to fibrous dust); and the hypothesis that, whereas overload of clearance can take place at high lung burdens after exposure has ceased, it is cancelled by the sustained stimulus to clearance mechanisms provided by the continuous challenge of chronic exposure. The linearity of the increase in lung burden is explained in terms of a kinetic model involving sequestration of some inhaled material to parts of the lung where it is difficult to clear. The particular sequestration model favoured is one where, the longer a particle remains in the lung without being cleared, the more likely it will be sequestrated (and therefore less likely cleared). It is believed that such ideas may eventually be useful in forming exposure-dose relations for epidemiology.


Immune mechanisms in leukemia: suppression of cellular immunity by drugs and x-irradiation.
The relative suppressive effects of x-irradiation (XR), cyclophosphamide (CY), prednisolone (PRD), and methotrexate (MTX) on the primary and secondary cellular immune response of C58/wm mice to syngeneic line Ib transplantable leukemia (Ib cells) were quantified. An LD10 dose of each agent was used for immunosuppression. XR, CY, and PRD were markedly suppressive for the primary immune response if given 24 hr before mice were immunized to Ib cells but less immunosuppressive if given 24 hr later. MTX was only slightly immunosuppressive XR, CY, and PRD also suppressed the secondary immune response if given before but not after antigen. The immunosuppressive effect of these agents was evaluated by defining their median immunosuppressive dose or the median time in days required for mice to recover from graded doses of each immunosuppressive agent. For example, the median recovery time from an LD10 of XR, CY, and PRD was 29.3, 19.7, and 3.7 days, respectively. Immunologic competence remaining after XR or drug treatment was quantified in terms of the LD50 dose of Ib cells required to kill recipient mice. For XR, CY, PRD, and MTX it was 10(6.16), 10(2.15), 10(6.90) and greater than 10(7.0) viable Ib cells, respectively. The overall results provided evidence that the primary and secondary cellular immune responses to a weak syngeneic tumor antigen were resistant to immunosuppression once they were initiated. There was a good correlation between the relative immunosuppressive effect of the test agents and the amount that they reduced the number of immune spleen cells. The agents also impaired the immunocompetence of individual spleen cells. Mechanisms by which XR or drugs might exert their immunosuppressive effects were discussed.


Optic Nerve Sheath Diameter is not Related to Intracranial Pressure in Subarachnoid Hemorrhage Patients.
Intracranial pressure (ICP) monitoring is essential after subarachnoid hemorrhage (SAH) to prevent secondary brain insults and to tailor individualized treatments. Optic nerve sheath diameter (ONSD), measured using ultrasound (US), could serve as a noninvasive bedside tool to estimate ICP, avoiding the risks of hemorrhage or infection related to intracranial catheters. The aims of this study were twofold: first, to explore the reliability of US for measuring ONSD; second, to establish whether the US-ONSD can be considered a proxy for ICP in SAH patients early after bleeding. For the first aim, we compared the ONSD measurements given by magnetic resonance imaging (MRI-ONSD) with the US-ONSD findings. For the second aim, we analyzed the relationship between US-ONSD measurements and ICP values. Adult patients with diagnosis of aneurysmal SAH and external ventricular drainage system (EVD) were included. Ten patients were examined by MRI to assess ONSD, and the results were compared to the diameter given by US. In 20 patients, the US-ONSD values were related to ICP measured simultaneously through EVD. In ten of these patients, we explored the changes in the US-ONSD at the time of controlled and fairly rapid changes in ICP after cerebrospinal fluid (CSF) drainage. US-ONSD measurements at the bedside were accurate, very similar to the diameters measured by MRI (the mean difference in the Bland-Altman plot was 0.08 mm, 95% limits of agreement: - 1.13; + 1.23 mm). No clear relationship was detectable between the ICP and US-ONSD, and a linear regression model showed an angular coefficient very close to 0 (p > 0.05). US-ONSD and ICP values were in agreement after CSF drainage and shifts in ICP in a limited number of patients. US-ONSD measurement does not accurately estimate ICP in SAH patients in the intensive care unit.


A membrane-associated Mn-superoxide dismutase protects the photosynthetic apparatus and nitrogenase from oxidative damage in the Cyanobacterium Anabaena sp. PCC 7120.
We investigated the functions of a membrane-associated manganese superoxide dismutase (MnSOD) of the heterocystous cyanobacterium Anabaena sp. PCC 7120. The gene sodA encoding MnSOD was inactivated by interposon mutagenesis and it was confirmed by Southern hybridization and immunoblotting. The strain A17, lacking sodA, grew more slowly than the wild type, and the difference in growth rates between the two strains became larger with an increase in growth light intensity. More severe inhibition of growth of A17 was observed when the cells were grown in the absence of combined nitrogen. Complementation of A17 with a full copy of the sodA gene restored the wild-type phenotypes. Strain A17 produced more malondialdehyde than did the wild type, especially under high light intensity, indicating more lipid peroxidation in the absence of MnSOD. A17 was also more susceptible to photoinhibition by a high light, and it was shown that both PSII and PSI were more severely damaged by the photoinhibitory light in A17, suggesting that the MnSOD plays important roles in protection of both photosystems. Immunoblotting revealed that the MnSOD was present in vegetative cells and heterocysts. Light greatly stimulated nitrogenase activity in the wild type under both aerobic and anaerobic conditions, but stimulated nitrogenase activity in A17 only slightly in air. The results suggest that reactive oxygen species produced in heterocysts under aerobic conditions cause the inactivation of nitrogenase in the absence of MnSOD.


Transmission properties of rectangular hollow waveguides for CO(2) laser light.
Transmission losses of two types of dielectric-coated rectangular waveguide are analyzed theoretically to transmit linearly or circularly polarized CO(2) laser light. When ZnS and PbTe are chosen as coating materials, waveguide losses are reduced by ~ 1 order of magnitude by increasing each pair of coating materials. Phosphor bronze rectangular waveguides with small aspect ratios and various cross sections, whose inner walls are coated with a single PbF(2) layer, are fabricated by using vacuum-evaporation and assembly techniques. Transmission losses of straight and bent waveguides are measured for coherent CO(2) laser light as well as for incoherent infrared lights. Straight waveguide losses of 0.1 dB/m including a coupling loss from a laser, are obtained at a 10.6-microm wavelength for 1-m-long and small-core waveguides.


A Review of Extraction and Analysis of Bioactives in Oat and Barley and Scope for Use of Novel Food Processing Technologies.
Oat and barely are cereal crops mainly used as animal feed and for the purposes of malting and brewing, respectively. Some studies have indicated that consumption of oat and barley rich foods may reduce the risk of some chronic diseases such as coronary heart disease, type II diabetes and cancer. Whilst there is no absolute consensus, some of these benefits may be linked to presence of compounds such as phenolics, vitamin E and β-glucan in these cereals. A number of benefits have also been linked to the lipid component (sterols, fatty acids) and the proteins and bioactive peptides in oats and barley. Since the available evidence is pointing toward the possible health benefits of oat and barley components, a number of authors have examined techniques for recovering them from their native sources. In the present review, we summarise and examine the range of conventional techniques that have been used for the purpose of extraction and detection of these bioactives. In addition, the recent advances in use of novel food processing technologies as a substitute to conventional processes for extraction of bioactives from oats and barley, has been discussed.


Molecular basis of beta-galactosidase alpha-complementation.
In previous studies, a cyanogen bromide peptide derived from amino-acid residues 3-92 of beta-galactosidase (EC 18.104.22.168; beta-D-galactoside galactohydrolase) was shown to have alpha-donor activity in intracistronic alpha-complementation. We have now isolated the defective beta-galactosidase alpha-acceptor protein from the deletion mutant strain M15 of Escherichia coli and find that it lacks residues 11-41 of betal-galactosidase. This is demonstrated by the isolation and sequence determination of a cyanogen bromide peptide from the M15 protein, which is identical to the corresponding peptide from beta-galactosidase except for the missing amino acids. We conclude that the alpha-donor peptide restores the region missing in the M15 protein.


A second failure to replicate the Gottman et al. (1995) typology of men who abuse intimate partners...and possible reasons why.
The present study attempts to replicate Gottman et al.'s (1995; see record 1995-44075-001; and Jacobson & Gottman, 1998) psychophysiological study that classifies partner assaultive men into two distinct groups: heart rate (HR) decelerators (Type 1 batterers) and HR accelerators (Type 2 batterers). Current results indicate no significant differences between Type 1 and Type 2 batterers on the antisocial spectrum of behaviors. Resting HR, rather than HR change, was negatively related to the antisocial spectrum of behavior for batterers with severe, clinical levels of violence only. Reasons for subsequent failures to replicate the Type 1 versus Type 2 distinctions may be attributable to unusually high autonomic arousal during baseline in the original study. Consideration of resting HR and the use of dimensional as opposed to categorical approaches in analyzing the heterogeneity of batterers are proposed as possible solutions to clarifying inconsistencies across laboratories.


Murine coronaviruses: isolation and characterization of two plaque morphology variants of the JHM neurotropic strain.
Two plaque-size variants of the neurotropic JHM strain of mouse hepatitis virus have been isolated from the virus stock after eight serial passages in suckling mouse brain. One variant, JHM-DL, produces large plaques, while the other, JHM-DS, produces small plaques in tissue culture. DS replicates more slowly, has a lower virus yield in vitro, and is less virulent for mice than DL. They also differ in their pathogenicity for mice: JHM-DL infection results in acute encephalomyelitis while JHM-DS infection results in demyelination. Oligonucleotide fingerprint analysis of the RNA genomes of these two variants revealed that they had almost identical genetic sequences. Each variant, however, had a unique oligonucleotide spot not found in the other. The unique spot of the large plaque variant, JHM-DL, was localized at approximately 3 to 5 kb from the 3' end, while the JHM-DS unique spot was mapped at 14 to 15 kb from the 3' end of the genome. We have further shown that these oligonucleotide changes are not correlated with the plaque morphology. These two viruses may be useful for studying the molecular basis of virus-induced demyelination.


Soy glycinin: influence of pH and ionic strength on solubility and molecular structure at ambient temperatures.
This study describes the relationship between the solubility of glycinin, a major soy protein, and its structural properties at a quaternary, tertiary, and secondary folding level under conditions representative for food products. When the ionic strength is lowered from 0.5 to 0.2 or 0.03, the basic polypeptides shift more to the exterior of the glycinin complex, as determined at pH 7.6 by labeling solvent-exposed lysines, supported by the study of the proteolytic action of clostripain on glycinin. This structural reorganization caused the pH of minimal solubility to shift to higher values. Ultracentrifugational analysis shows that at pH 7.6 and an ionic strength of 0.5 glycinin forms hexameric complexes (11S), whereas at pH 3.8 and at an ionic strength of 0.03 glycinin exists as trimers (7S). Intermediate situations are obtained by modulation of pH and ionic strength. The observed quaternary dissociation correlates with an increased amount of nonstructured protein at a secondary level and with changes in tertiary folding as determined using circular dichroism. Tryptophan fluorescence shows no significant structural changes for different ionic strengths but demonstrates a more tightly packed fluorophore environment when the pH is lowered from 7.6 to 3.8.


bicoid mRNA localization signal: phylogenetic conservation of function and RNA secondary structure.
Transcripts of the bicoid (bcd) gene are localized to the anterior pole of the Drosophila oocyte, thereby allowing formation in the embryo of an anteroposterior gradient of the bcd protein morphogen. We previously showed that a 630 nucleotide portion of the 3' noncoding region of the bcd mRNA is necessary for this localization, and is sufficient to confer anterior localization on a heterologous transcript. Here I have used a comparative analysis to begin to more precisely define the cis-acting mRNA localization signal. The bcd genes from six additional Drosophila species were cloned, and DNA of the 3' noncoding regions sequenced. Three of these regions were tested interspecifically for mRNA localization in D. melanogaster and each functioned correctly; these regions must therefore contain the cis-acting signal. The primary sequences, which are up to 50% divergent from the D. melanogaster gene, show patchy homology throughout most of the region. Interestingly, all seven species can potentially form a large stereotypic secondary structure. This structure is a likely candidate for the localization signal and can be used for the rational design of mutations to test that possibility.


Classification of endometriosis.
The classification of endometriosis has evolved during this century under the influence of many factors. Although classifications initially paralleled the staging of pelvic malignancies, more modern systems follow the natural progression of this pathologic process. The diagnosis of endometriosis at earlier stages has been limited by the technology of the era. Previously, only patients with more severe disease or symptoms were evaluated and treated. Earlier therapies were more radical and centered on pain relief achieved with removal of the internal genitalia. The objectives of classification were to provide standardized documentation of the disease and--it was hoped--define those patients amenable to less radical therapy. As our diagnostic and therapeutic capabilities grew, so did our interest in infertile women with endometriosis and the treatment of such patients. The diagnosis of endometriosis was often made at an earlier stage of involvement, and therapy stressed repair rather than removal. Description of pelvic involvement, has become more detailed and accurate, and the stage of disease has become prognostic of future fertility. The basic tenet of the R-AFS classification system is to provide an objective and universal method for describing the severity of disease visualized at laparotomy or laparoscopy. Stage of disease will aid the physician in choosing the therapy most suited for the individual patient. If reparative surgery is undertaken, the stage of disease will reflect the extent of pelvic reconstruction required. Quantifying disease severity will, it is hoped, make it predictive of success in conception and prognostic of the need for future medical or surgical treatment. It will also serve to stratify patients with a similar degree and distribution of disease into somewhat homogeneous groups. This will permit reasonable comparisons of effectiveness of different therapies and investigators' methods. Refinement of this type of classification scheme will depend on critical analysis of the system's prognostic capabilities. Further studies will help elucidate the relative importance of individual variables on the potential for success in achieving pregnancy.


The combination of 9p21.3 genotype and biomarker profile improves a peripheral artery disease risk prediction model.
Peripheral artery disease (PAD) is a highly morbid condition affecting more than 8 million Americans. Frequently, PAD patients are unrecognized and therefore do not receive appropriate therapies. Therefore, new methods to identify PAD have been pursued, but have thus far had only modest success. Here we describe a new approach combining genomic and metabolic information to enhance the diagnosis of PAD. We measured the genotype of the chromosome 9p21 cardiovascular-risk polymorphism rs10757269 as well as the biomarkers C-reactive protein, cystatin C, β2-microglobulin, and plasma glucose in a study population of 393 patients undergoing coronary angiography. The rs10757269 allele was associated with PAD status (ankle-brachial index < 0.9) independent of biomarkers and traditional cardiovascular risk factors (odds ratio = 1.92; 95% confidence interval, 1.29-2.85). Importantly, compared to a previously validated risk factor-based PAD prediction model, the addition of biomarkers and rs10757269 significantly and incrementally improved PAD risk prediction as assessed by the net reclassification index (NRI = 33.5%; p = 0.001) and integrated discrimination improvement (IDI = 0.016; p = 0.017). In conclusion, a model including a panel of biomarkers, which includes both genomic information (which is reflective of heritable risk) and metabolic information (which integrates environmental exposures), predicts the presence or absence of PAD better than established risk models, suggesting clinical utility for the diagnosis of PAD.


Laboratory and Field Evaluation of Host-Related Foraging Odor-Cue Combinations to Attract Drosophila suzukii (Diptera: Drosophilidae).
The invasive spotted-wing drosophila, Drosophila suzukii (Matsumura), is a major pest of soft-skinned fruits. Since its introduction into North America and Europe, significant progress has been made in understanding the volatile cues used by this fly during food, oviposition site, and mate finding. Despite this progress, commercially available lures are non-selective. Here, we tested two Hanseniaspora uvarum (Niehaus) yeast compounds (isoamyl acetate and isobutyl acetate) and a leaf compound β-cyclocitral alone and in combination with a blend of four fermentation compounds ('Fermentation lure': acetic acid, ethanol, methionol, and acetoin) to improve D. suzukii attraction and selectivity. In laboratory assays, males and females were attracted to all seven individual compounds, although in electrophysiological assays, their antennae exhibited a dose-dependent response to only four of these compounds. In two-choice cage studies, the Fermentation lure was more attractive to D. suzukii than water controls, whereas β-cyclocitral and the mixture of isoamyl acetate and isobutyl acetate were not attractive in this larger-cage study. Moreover, adding the two-component H. uvarum compound blend to the Fermentation lure reduced D. suzukii attraction to the Fermentation blend. When these experiments were repeated in blueberry, raspberry, blackberry, and cherry orchards across several states in the United States over 2 yr, similar outcomes were observed: β-cyclocitral or the mixture of the H. uvarum blend did not improve the attractiveness of the Fermentation lure or its selectivity. This study demonstrates that cues from different sources may interfere with each other and reduce D. suzukii attraction to otherwise attractive odor combinations.


The phytological future of prostate cancer staging: PSMA-PET and the dandelion theory.
PSMA-PET is changing how we stage prostate cancer, both in the primary setting and with relapse after treatment. It allows us to identify lesions in the bones and lymph nodes that were not previously visible on conventional imaging with bone scan and CT/MRI. In this Special Report we review the 'state of the art' for PSMA imaging and discuss the implications for treatment decisions in prostate cancer. We liken early high risk or metastatic prostate to a common phytological problem: the dandelion. In this analogous situation, we consider the additional evidence needed for us to start plucking out the original dandelion and for us to focus attention on killing the seeded weeds that are identifiable elsewhere in the lawn.


Pediatric HIV+ Kaposi sarcoma exhibits clinical, virological, and molecular features different from the adult disease.
BACKGROUNDKaposi sarcoma (KS) is among the most common childhood cancers in Eastern and Central Africa. Pediatric KS has a distinctive clinical presentation compared with adult KS, which includes a tendency for primary lymph node involvement, a considerable proportion of patients lacking cutaneous lesions, and a potential for fulminant disease. The molecular mechanisms or correlates for these disease features are unknown.METHODSThis was a cross-sectional study. All cases were confirmed by IHC for KS-associated herpesvirus (KSHV) LANA protein. Baseline blood samples were profiled for HIV and KSHV genome copy numbers by qPCR and secreted cytokines by ELISA. Biopsies were characterized for viral and human transcription, and KSHV genomes were determined when possible.RESULTSSeventy participants with pediatric KS were enrolled between June 2013 and August 2019 in Malawi and compared with adult patients with KS. They exhibited high KSHV genome copy numbers and IL-6/IL-10 levels. Four biopsies (16%) had a viral transcription pattern consistent with lytic viral replication.CONCLUSIONThe unique features of pediatric KS may contribute to the specific clinical manifestations and may direct future treatment options.FUNDINGUS National Institutes of Health U54-CA-254569, PO1-CA019014, U54-CA254564, RO1-CA23958.


Is serum leptin level regulated by thyroid functions, lipid metabolism and insulin resistance in polycystic ovary syndrome?
Our aim was to determine whether serum leptin level is regulated by thyroid hormones, lipid metabolic products and insulin resistance status in women with polycystic ovary syndrome (PCOS). A prospective case-controlled study was carried out in Istanbul University, Cerrahpasa School of Medicine in 25 lean PCOS (L-PCOS) women, 19 obese PCOS (O-PCOS) women and 28 normal women. The diagnosis of PCOS was established according to the clinical, hormonal (elevated luteinizing hormone and serum androgens) and ultrasonographic findings. Fasting serum levels of thyroid stimulating hormone (TSH), free triiodothyronine (FT3), free thyroxine (FT4), fasting glucose, insulin, total cholesterol (TC), triglyceride (TG), high-density lipoprotein-cholesterol (HDL-C), low-density lipoprotein-cholesterol (LDL-C), very low-density lipoprotein-cholesterol (VLDL-C) and leptin were measured and compared in the three groups and the correlations between serum levels of leptin and other parameters were evaluated. Serum leptin levels were higher in the O-PCOS group, while its level was comparable between the L-PCOS and control groups. Serum levels of FT4 were significantly lower in both L-PCOS and O-PCOS groups than the control group. Women in both L-PCOS and O-PCOS groups were found to be significantly hyperinsulinemic and insulin resistant. Serum levels of TC, VLDL-C and TG were significantly higher in the O-PCOS group, while serum HDL-C level was lower. There was a poor correlation between serum leptin, and FT4, TC, TG, HDL-C and VLDL-C levels. A significant correlation was observed between serum leptin levels and both BMI and insulin resistance status in PCOS. We believe that, although thyroid hormones and lipid metabolic products do not seem to participate in the regulation of serum leptin levels, BMI and insulin resistance status may have a key role in women with PCOS.


Stereotaxic injection of IgG from patients with Alzheimer disease initiates injury of cholinergic neurons of the basal forebrain.
The participation of an immune/inflammatory process in the pathomechanism of sporadic Alzheimer disease (AD) has been suggested by evidence for activated microglia and the potential therapeutic benefit of anti-inflammatory medication. To define a possible role for IgG in the immune/inflammatory process of AD in humans, we assayed the ability of IgG samples from patients with AD to target the injury to cholinergic neurons in rat basal forebrain in vivo. IgG purified from the serum or plasma from patients with AD and patients with other neurological disease who were used as control (DC) patients was injected stereotaxically into the medial septum of adult rats. Four weeks later coronal sections of the whole medial septum-diagonal bands of Broca region were immunostained for choline acetyltransferase (ChAT) to identify cholinergic neuronal cells. University medical centers. Blood samples were collected from 8 patients with probable and definite AD and from 6 age-matched DC patients. Detection of changes in the number of ChAT immunopositive cell profiles in sections and statistical evaluation. Four weeks after the injections, IgG samples from patients with AD significantly reduced the number of ChAT-immunostained cell profiles in the whole medial septum-diagonal bands of Broca region compared with IgGs from DC patients. Neither DC IgGs nor saline solution significantly decreased the number of ChAT-immunopositive neuronal cell profiles. Data document that IgG from patients with AD can target a stereotaxically induced immune/inflammatory injury to cholinergic neurons in the rat basal forebrain in vivo.


Complex Formation of Lead(II) with Nucleotides and Their Constituents.
Lead is widely distributed in the environment; it is known to mankind for thousands of years and its toxicity is nowadays (again) well recognized, though on the molecular level only partly understood. One of the reasons for this shortcoming is that the coordination chemistry of the biologically important lead(II) is complicated due to the various coordination numbers it can adopt (CN = 4 to 10) as well as by the 6s2 electron lone pair which, with CN = 4, can shield one side of the Pb2+ coordination sphere. The chapter focuses on the properties of Pb2+ complexes formed with nucleotides and their constituents and derivatives. Covered are (among others) the complexes formed with hydroxy groups and sugar residues, the interactions with the various nucleobases occurring in nucleic acids, as well as complexes of phosphates. It is expeced that such interactions, next to those like with lipids and proteins, are responsible for the toxic properties of lead. To emphasize the special properties of Pb2+ complexes, these are compared as far as possible with the corresponding properties of the Ca2+, Fe2+, Cu2+, Zn2+, and Cd2+ species. It needs to be mentioned that the hard-soft rule fails with Pb2+. This metal ion forms complexes with ligands offering O donors of a stability comparable to that of Cu2+. In contrast, with aromatic N ligands, like imidazole or N7 sites of purines, complex stability is comparable to that of the corresponding Fe2+ complexes. The properties of Pb2+ towards S donor sites are difficult to generalize: On the one hand Pb2+ forms very stable complexes with nucleoside 5'-O-thiomonophosphates by coordinating to nearly 100% at S in the thiophosphate group; however, on the other hand, once a sulfur atom replaces one of the terminal oxygen atoms in the phosphodiester linkage, macrochelate formation of the phosphate-bound Pb2+ occurs with the O and not the S site. Quite generally, the phosphodiester linkage is a relatively weak binding site, but the affinity increases further to the mono- and then to the di- and triphosphate. The same holds for the corresponding nucleotides, though the Pb2+ affinity had to be estimated via that of the Cu2+ complexes for some of these ligands. Complex stability of the pyrimidine-nucleotides (due to their anti conformation) is solely determined by the coordinating tendency of the phosphate group(s); this also holds for the Pb2+ complex of adenosine 5'-monophosphate. For the other purinenucleotides macrochelate formation takes place by the interaction of the phosphate-coordinated Pb2+ with the N7/(C6)O site of, e.g., the guanine residue. The extents of the formation degrees of these chelates are summarized. Unfortunately, information about mixed ligand (ternary) or other higher order comlexes is missing, but still it is hoped that the present overview will help to understand the interaction of Pb2+ with nucleotides and nucleic acids, and especially that it will facilitate further research in this fascinating area.


Population psychiatric medication prescription rates following a terrorist attack.
While several population-based studies have documented behavioral health disturbances following terrorist attacks, a number of mental health service utilization analyses present conflicting conclusions. The purpose of this study was to determine if mental health service utilization increased following a terrorist attack by assessing changes in psychoactive drug prescription rates. The rate of selective serotonin reuptake inhibitor (SSRI) prescriptions was measured among New York State Medicaid enrollees before and after the terrorist attacks of 11 September 2001. The association between geographic proximity to the events and changes in the rate of SSRI prescriptions around 11 September 2001 was assessed. From September to December 2001, among individuals residing within three miles of the World Trade Center site, there was an 18.2% increase in the SSRI prescription rate compared to the previous eight-month period (p = 0.0011). While there was a 9.3% increase for non-New York City residents, this change was not statistically significant (p = 0.74). There was a quantifiable increase in the dispensing of psychoactive drugs following the terrorist attacks of 11 September 2001, and this effect varied by geographic proximity to the events. These findings build on the growing body of knowledge on the pervasive effects of disasters and terrorist events for population health, and demonstrate the need to include mental and behavioral health as key components of surge capacity and public health response to mass traumas.


Diaryl hydrazones as multifunctional inhibitors of amyloid self-assembly.
The design and application of an effective, new class of multifunctional small molecule inhibitors of amyloid self-assembly are described. Several compounds based on the diaryl hydrazone scaffold were designed. Forty-four substituted derivatives of this core structure were synthesized using a variety of benzaldehydes and phenylhydrazines and characterized. The inhibitor candidates were evaluated in multiple assays, including the inhibition of amyloid β (Aβ) fibrillogenesis and oligomer formation and the reverse processes, the disassembly of preformed fibrils and oligomers. Because the structure of the hydrazone-based inhibitors mimics the redox features of the antioxidant resveratrol, the radical scavenging effect of the compounds was evaluated by colorimetric assays against 2,2-diphenyl-1-picrylhydrazyl and superoxide radicals. The hydrazone scaffold was active in all of the different assays. The structure-activity relationship revealed that the substituents on the aromatic rings had a considerable effect on the overall activity of the compounds. The inhibitors showed strong activity in fibrillogenesis inhibition and disassembly, and even greater potency in the inhibition of oligomer formation and oligomer disassembly. Supporting the quantitative fluorometric and colorimetric assays, size exclusion chromatographic studies indicated that the best compounds practically eliminated or substantially inhibited the formation of soluble, aggregated Aβ species, as well. Atomic force microscopy was also applied to monitor the morphology of Aβ deposits. The compounds also possessed the predicted antioxidant properties; approximately 30% of the synthesized compounds showed a radical scavenging effect equal to or better than that of resveratrol or ascorbic acid.


Crystallization and prevention of supercooling of microencapsulated n-alkanes.
Microencapsulated n-alkanes (n-octadecane, n-nonadecane, and n-eicosane) were synthesized by in situ polymerization using urea-melamine-formaldehyde polymer as shells. Microcapsules 5.0 and 10.0 wt% of 1-tetradecanol, paraffin, and 1-octadecanol were used as nucleating agents. The fabrication was characterized using Fourier transform infrared, light microscopy, and scanning electron microscopy. The crystallization and prevention of supercooling of the microcapsules are studied using differential scanning calorimetry (DSC) and wide-angle X-ray diffraction. The crystal system of the microencapsulated n-alkane is the same as that of the bulk. The enthalpies of the microcapsules containing 70 wt% n-alkanes are approximately 160 J/g. The melting temperature of the n-alkanes in the microcapsule is the same as that in the bulk. There are multiple peaks on the DSC cooling curves that are attributed to liquid-rotator, rotator-crystal, and liquid-crystal transitions. The DSC cooling behavior of microencapsulated n-octadecane is affected by the average diameters. The measured maximum degree of supercooling of the microencapsulated n-octadecane is approximately 26.0 degrees C at a heating and cooling rate of 10.0 degrees C/min. The degree of supercooling of microencapsulated n-octadecane is decreased by adding 10.0 wt% of 1-octadecanol as a nucleating agent.


Influences on clinical practice: the case of glue ear.
A case study of clinical practice in children with glue ear is presented. The case is part of a larger project, funded by the North Thames Research and Development Programme, that sought to explore the part played by clinicians in the implementation of research and development into practice in two areas: adult asthma and glue ear in children. What is striking about this case is the differences found in every area of the analysis. That is, diversity was found in views about diagnosis and treatment of glue ear; the organisation of related services; and in the reported practice of our interviewees, both between particular groupings of clinical staff and within these groupings. The challenge inherent in the case is to go beyond describing the complexity and differences that were found, and look for patterns in the accounts of practice and tease out why such patterns may occur.


End-binding proteins EB3 and EB1 link microtubules to ankyrin G in the axon initial segment.
The axon initial segment (AIS) plays a key role in maintaining the molecular and functional polarity of the neuron. The relationship between the AIS architecture and the microtubules (MTs) supporting axonal transport is unknown. Here we provide evidence that the MT plus-end-binding (EB) proteins EB1 and EB3 have a role in the AIS in addition to their MT plus-end tracking protein behavior in other neuronal compartments. In mature neurons, EB3 is concentrated and stabilized in the AIS. We identified a direct interaction between EB3/EB1 and the AIS scaffold protein ankyrin G (ankG). In addition, EB3 and EB1 participate in AIS maintenance, and AIS disassembly through ankG knockdown leads to cell-wide up-regulation of EB3 and EB1 comets. Thus, EB3 and EB1 coordinate a molecular and functional interplay between ankG and the AIS MTs that supports the central role of ankG in the maintenance of neuronal polarity.


Using the Practical, Robust Implementation and Sustainability Model (PRISM) to qualitatively assess multilevel contextual factors to help plan, implement, evaluate, and disseminate health services programs.
There is consensus in dissemination and implementation (D&I) science that addressing contextual factors is critically important for understanding translation of health care delivery interventions but little agreement on which contextual factors are key determinants of implementation outcomes. We describe the application of the Practical Robust Implementation and Sustainability Model (PRISM), which expands the Reach, Effectiveness, Adoption, Implementation, Maintenance (RE-AIM) framework to identify contextual factors across four diverse programs. Multiple qualitative methods were used to collect multilevel, multistakeholder perspectives from the adopting organizations and staff. We identified measures for evaluating context through the various domains of PRISM to guide health services research across the phases of program implementation. The PRISM domains of Recipients, Implementation and Sustainability Infrastructure, and External Environment identified important multilevel contextual factors, including variability in operational processes and available resources. These domains helped to facilitate planning and implementation phases of the four interventions and guide purposeful adaptations. We found assessments of PRISM domains useful to systematically assess multilevel contextual factors across various content areas as well as phases of program implementation. Additionally, these contextual factors were found to be relevant to RE-AIM outcomes. Lessons learned can be applied to future research as there is a need to investigate the measurement properties of PRISM and continue to test which contextual factors are most important to successful implementation and for which outcomes.


Th22 cells as well as Th17 cells expand differentially in patients with early-stage and late-stage myelodysplastic syndrome.
Immunological mechanisms are increasingly recognized in the progression of myelodysplastic syndrome (MDS). Early-stage MDS (E-MDS) is characterized by autoimmune-mediated myelosuppression whereas late-stage MDS (L-MDS) involves immune evasion, giving dysplastic cells growth potential to progress into acute myeloid leukemia. T-helper (Th) 22 is involved in the pathogenesis of inflammatory autoimmunity and tumorigenesis. The roles of Th22 cells in the pathophysiology of E-MDS and L-MDS remain unsettled. We studied 37 MDS patients (E-MDS, n = 17; L-MDS, n = 20) and 20 healthy controls to characterize their peripheral blood (PB), as well as 25 MDS patients and 10 healthy controls to characterize their bone marrow(BM). The expression of Interleukin-22 (IL-22), IL-17 or interferon gamma (IFN-γ) was examined in E-MDS, L-MDS patients and controls by flow cytometry. The mRNA expression levels of RAR-related orphan receptor C (RORC), IL-6, tumor necrosis factor alpha (TNF-α) and IL-23 in peripheral blood mononuclear cells (PBMCs) were determined by real-time quantitative polymerase chain reaction. The levels of IL-22 and IL-17 both in PB and BM plasma were examined by enzyme-linked immunosorbent assay. In E-MDS, peripheral Th17 cells were significantly elevated and correlated with peripheral Th22 cells compared with healthy controls and L-MDS. Significantly higher levels of peripheral Th22 expansion, mRNA expression of IL-6, TNF-α and lower level of RORC mRNA expression were observed in L-MDS compared with E-MDS. No statistical difference was found in IL-23 mRNA expression or plasma IL-22, IL-17 levels among E-MDS, L-MDS and controls. Our data demonstrated that L-MDS cohort had increased frequencies of peripheral Th22 cells and higher mRNA expression levels of IL-6 and TNF-α, indicating that Th22 cells along with Th17 cells or not are involved in the dynamic immune responses of MDS.


[Prostitution and AIDS in the city of Mexico].
Since 1987 The National AIDS Council in Mexico (CONASIDA), has conducted research projects among female commercial sex workers (CSW), who are seen at the AIDS Information Centers. Through these studies, we have been able to find out how different issues influence commercial sex work in Mexico City. Since the beginning of our own studies in 1989, HIV prevalence in this particular group has remained low (0.04%-0.2%). Factors found to be associated with condom use were: educational level, not having children, more experience in sex work and younger age. Condom use was reported by 88% of the women; however, qualitative information and high STD prevalence rates suggest a much lower condom use. Mandatory HIV testing is not useful as a preventive measure among sex workers. Persuasive and structural interventions must be implemented in accordance with local and legal situations.


During development of rheumatoid arthritis, intermetatarsal bursitis may occur before clinical joint swelling: a large imaging study in patients with clinically suspect arthralgia.
Intermetatarsal bursitis (IMB) represents juxta-articular synovial inflammation of the intermetatarsal bursae. Recent MRI studies identified IMB as feature of early RA, but whether IMB already occurs in the pre-arthritic phase is unknown. We performed a large MRI study in clinically suspect arthralgia (CSA) to assess the occurrence and prognostic value of IMB. A total of 577 consecutive CSA patients underwent contrast-enhanced MRI of the forefoot, metacarpophalangeal joints and wrist. MRIs were evaluated for subclinical synovitis/tenosynovitis/osteitis in line with the RA MRI scoring system (summed as RAMRIS inflammation) and for IMB. IMB was considered present if uncommon in the general population at the same location (i.e. size scored above the 95th percentile in age-matched symptom-free controls). The relation of IMB with other MRI-detected subclinical inflammation (synovitis/tenosynovitis/osteitis) was studied. Cox-regression assessed the association with clinical arthritis development during median 25 months follow-up. ACPA stratification was performed. At presentation with CSA, 23% had IMB. IMB was more frequent in ACPA-positive than ACPA-negative CSA (47% vs 19%, P < 0.001). Patients with IMB were more likely to also have subclinical synovitis [OR 3.4 (95% CI 1.8, 6.5)] and tenosynovitis [5.9(2.8, 12.6)]. IMB conferred higher risk of developing arthritis [HR 1.6(1.0-2.7) adjusted for other subclinical inflammation]. IMB-presence predicted arthritis development in ACPA-positive CSA [adjusted HR 2.2(1.0-4.7)], but not in ACPA-negative CSA-patients [0.8(0.4-1.7)]. Approximately a quarter of CSA patients have IMB, which is frequently accompanied by subclinical synovitis and tenosynovitis. IMB precedes development of clinical arthritis, particularly in ACPA-positive CSA. These results reinforce the notion that juxta-articular synovial inflammation is involved in the earliest phases of RA development.


Antimicrobial Stewardship for Outpatients with Chronic Bone and Joint Infections in the Orthopaedic Clinic of an Academic Tertiary Hospital, South Africa.
Bone and joint infections are associated with prolonged hospitalizations, high morbidity and complexity of care. They are difficult to treat, and successful therapy requires organism-specific antimicrobial therapy at high doses for a prolonged duration as recommended in standard treatment guidelines (STGs). Adherence to the treatment plan is equally important, which is enhanced with knowledge of the condition as well as appropriate antibiotics. Consequently, the aim of this study was to provide antimicrobial stewardship (AMS) services to outpatients with chronic bone and joint infections presenting to the orthopaedic clinic at a public South African tertiary hospital. A total of 44 patients participated in this study. Chronic osteomyelitis was diagnosed in 39 (89%) patients and septic arthritis in 5 (11%). The majority (43%) of infections were caused by Staphylococcus aureus followed by Pseudomonas aeruginosa (14%). Seventy-one antibiotics were prescribed at baseline with rifampicin prescribed the most (39%), followed by ciprofloxacin (23%). The majority (96%) of the antibiotics were not prescribed according to the South African STG; however, interventions were only needed in 31% of prescribed antibiotics (n = 71) since the STG only recommends empiric therapy directed against Staphylococcus aureus. Seventy-seven percent of the patients obtained a high self-reported adherence score at baseline. Consequently, there is a need to improve AMS in bone and joint infections to improve future care.


Methods for parallel-detection-based ultrasound-modulated optical tomography.
The research reported here focuses on ultrasound-modulated optical tomography based on parallel speckle detection. Four methods were investigated for signal acquisition and analysis, in which laser speckle statistics was applied. The methods were compared with the previously used four-phase method in the imaging of all-biological-tissue samples, in which the buried objects were also biological tissues. The image quality obtained with these methods was comparable with that obtained with the four-phase method; in addition, these methods have advantages in reducing acquisition time and improving the signal-to-noise ratio.


Localization of pain and self-reported rape in a female community sample.
Studies suggest that rape increases risk of medically unexplained pain in women. At present it is not clear whether rape is associated with pain at specific locations or at multiple locations. In this study we tested the hypothesis that rape was associated with a preferential increase in risk of pelvic pain that was not explained by pain at other sites. We relied on an existing community study that oversampled women with fibromyalgia and major depression. Localization was assessed by asking about pain at four sites: pelvic region; jaw/face; headache; and lower back. Three groups were identified using a structured telephone interview: Abuse Only (sexual/physical abuse excluding rape); Rape+Abuse (rape in addition to other sexual/physical abuse); and No Abuse. Compared with the No Abuse group, the Rape+Abuse group was eight times more likely to have pelvic pain and 3.7 times more likely to have jaw/face pain after we controlled for the effect of widespread pain. Rape was not associated with lower back pain or headache. The Abuse Only group did not show a preferential increase in risk of pain at any of the four locations that were assessed. After controlling for pain at other locations, we found that the Rape + Abuse group was 10 times more likely to report pelvic pain than the No Abuse group (P<0.005). In accord with the localization hypothesis, self-reported rape was uniquely associated with pelvic pain. Future efforts to account for pain in the aftermath of rape must specify a mechanism that can simultaneously cause widespread pain as well as increase risk of localized pain.


Trajectories of Dating Violence Victimization and Perpetration among Rural Adolescents.
Research is inconclusive about the trajectory of dating violence during adolescence and whether there are differences across gender and race/ethnicity. We examined dating victimization and perpetration trajectories among a diverse sample of rural youth (N = 580, 52.7% female, 49% Black, 39% White, 11% Hispanic or other minorities) in middle and high school who were surveyed annually across four years and explored the influences of gender and ethnicity. The results based on cohort-sequential latent growth modeling revealed that for boys, victimization peaked at 11th grade, and then declined. For girls, victimization was stable throughout adolescence. Perpetration was reported less frequently and increased steadily for males and females. For White youth, victimization peaked at grades 9 and 10, followed by a decline. For Black youth, victimization followed a linear increase. Perpetration trajectory followed a linear increase for White and Black but not Hispanic youth. The findings indicate that the developmental progression of dating violence during adolescence varies by demographics. The discussion focuses on future directions for research on teen dating violence among rural youth and implications for prevention and interventions initiatives.


Enterotoxin production by coagulase-negative staphylococci in restaurant workers from Kuwait City may be a potential cause of food poisoning.
Staphylococcus aureus and coagulase-negative staphylococci (CNS) were isolated from the hands of food handlers in 50 restaurants in Kuwait City and studied for the production of staphylococcal enterotoxins, toxic shock syndrome toxin-1, slime and resistance to antimicrobial agents. One or a combination of staphylococcal enterotoxins A, B or C were produced by 6% of the isolates, with the majority producing enterotoxin B. Toxic shock syndrome toxin-1 was detected in c. 7% of the isolates; 47% produced slime. In all, 21% of the isolates were resistant to tetracycline and 11.2% were resistant to propamidine isethionate and mercuric chloride. There was no correlation between slime and toxin production or between slime production and antibiotic resistance. The detection of enterotoxigenic CNS on food handlers suggests that such strains may contribute to food poisoning if food is contaminated by them and held in conditions that allow their growth and elaboration of the enterotoxins. It is recommended that enterotoxigenic CNS should not be ignored when investigating suspected cases of staphylococcal food poisoning.


Direct administration of 2-Hydroxypropyl-Beta-Cyclodextrin into guinea pig cochleae: Effects on physiological and histological measurements.
2-Hydroxypropyl-Beta-Cyclodextrin (HPβCD) can be used to treat Niemann-Pick type C disease, Alzheimer's disease, and atherosclerosis. But, a consequence is that HPβCD can cause hearing loss. HPβCD was recently found to be toxic to outer hair cells (OHCs) in the organ of Corti. Previous studies on the chronic effects of in vivo HPβCD toxicity did not know the intra-cochlear concentration of HPβCD and attributed variable effects on OHCs to indirect drug delivery to the cochlea. We studied the acute effects of known HPβCD concentrations administered directly into intact guinea pig cochleae. Our novel approach injected solutions through pipette sealed into scala tympani in the cochlear apex. Solutions were driven along the length of the cochlear spiral toward the cochlear aqueduct in the base. This method ensured that therapeutic levels were achieved throughout the cochlea, including those regions tuned to mid to low frequencies and code speech vowels and background noise. A wide variety of measurements were made. Results were compared to measurements from ears treated with the HPβCD analog methyl-β-cyclodextrin (MβCD), salicylate that is well known to attenuate the gain of the cochlear amplifier, and injection of artificial perilymph alone (controls). Histological data showed that OHCs appeared normal after treatment with a low dose of HPβCD, and physiological data was consistent with attenuation of cochlear amplifier gain and disruption of non-linearity associated with transferring acoustic sound into neural excitation, an origin of distortion products that are commonly used to objectively assess hearing and hearing loss. A high dose of HPβCD caused sporadic OHC losses and markedly affected all physiologic measurements. MβCD caused virulent destruction of OHCs and physiologic responses. Toxicity of HPβCD to OHC along the cochlear length is variable even when a known intra-cochlear concentration is administered, at least for the duration of our acute studies.


Immunomodulatory Drugs: IMiDs in Acute Myeloid Leukemia (AML).
AML patients have an aberrant and dysfunctional immune state, paving the way for novel agents targeting pathways that integrate with immune signaling, function, and response. Small molecule immunomodulatory drugs (IMiDs) represent a class of agents derived from the parent compound, thalidomide. There are currently 3 IMiDs approved for a variety of malignancies: thalidomide, lenalidomide, and the newest agent, pomalidomide. IMiDs lead to a multitude of immunobiologic effects such as cytokine modulation, co-stimulation of T cells, down-regulation of co-inhibitory molecules, enhancing natural killer cell activity, inhibition of regulatory T cells, and repairing perturbed synapse formation on T cells. IMiDs have been extensively studied in various AML settings with promising clinical activity. This review discusses the immunologic effects of IMiDs, the rationale for studying IMiDs in AML, and the published and ongoing clinical trials investigating IMiD activity in AML.


Sensitivity analysis and potential uses of a novel gamma interferon release assay for diagnosis of tuberculosis.
Sputum smears for acid-fast bacilli (AFB) are the primary methods for diagnosis of tuberculosis (TB) in many countries. The tuberculin skin test (TST) is the primary method for diagnosis of latent TB infection (LTBI) worldwide. The poor sensitivity of the former and the poor specificity of the latter warrant the development of new tests and strategies to enhance diagnostic capabilities. We evaluated the sensitivity of an "in-tube" gamma interferon release assay (IGRA) using TB-specific antigens in comparison to the TST and the sputum smear for AFB in TB cases in South Africa. The sensitivity of the IGRA for TB was considered a surrogate of sensitivity in LTBI. Among 154 patients with a positive culture for Mycobacterium tuberculosis, the sensitivity of the IGRA for the diagnosis of TB varied by clinical subgroup from 64% to 82%, that of the TST varied from 85% to 94%, and that of two sputum smears for AFB varied from 35% to 53%. The sensitivity of the IGRA in human immunodeficiency virus (HIV)-infected TB cases was 81%. HIV-infected TB patients were significantly more likely to have indeterminate IGRA results and produced quantitatively less gamma interferon in response to TB-specific antigens than HIV-negative TB patients. The overall sensitivity of the TST in all TB cases was higher than that of the IGRA (90% versus 76%, respectively). The combined sensitivities of the TST plus IGRA and TST plus a single sputum smear were 96% and 93%, respectively. The TST combined with IGRA or with a single sputum smear may have a role in excluding the diagnosis of TB in some settings.


Crystal structure of a bacterial unsaturated glucuronyl hydrolase with specificity for heparin.
Extracellular matrix molecules such as glycosaminoglycans (GAGs) are typical targets for some pathogenic bacteria, which allow adherence to host cells. Bacterial polysaccharide lyases depolymerize GAGs in β-elimination reactions, and the resulting unsaturated disaccharides are subsequently degraded to constituent monosaccharides by unsaturated glucuronyl hydrolases (UGLs). UGL substrates are classified as 1,3- and 1,4-types based on the glycoside bonds. Unsaturated chondroitin and heparin disaccharides are typical members of 1,3- and 1,4-types, respectively. Here we show the reaction modes of bacterial UGLs with unsaturated heparin disaccharides by x-ray crystallography, docking simulation, and site-directed mutagenesis. Although streptococcal and Bacillus UGLs were active on unsaturated heparin disaccharides, those preferred 1,3- rather than 1,4-type substrates. The genome of GAG-degrading Pedobacter heparinus encodes 13 UGLs. Of these, Phep_2830 is known to be specific for unsaturated heparin disaccharides. The crystal structure of Phep_2830 was determined at 1.35-Å resolution. In comparison with structures of streptococcal and Bacillus UGLs, a pocket-like structure and lid loop at subsite +1 are characteristic of Phep_2830. Docking simulations of Phep_2830 with unsaturated heparin disaccharides demonstrated that the direction of substrate pyranose rings differs from that in unsaturated chondroitin disaccharides. Acetyl groups of unsaturated heparin disaccharides are well accommodated in the pocket at subsite +1, and aromatic residues of the lid loop are required for stacking interactions with substrates. Thus, site-directed mutations of the pocket and lid loop led to significantly reduced enzyme activity, suggesting that the pocket-like structure and lid loop are involved in the recognition of 1,4-type substrates by UGLs.


[Possible role of tRNA1Lys in recognition of pre-mRNA sequences involved in the control of splicing].
Previously we presented some evidence for involvement of the imported tRNA1Lys in splicing the mitochondrial mosaic genes transcripts in yeast. The new data provided here together with our previous results show that tRNA1Lys/transcripts complementarity permits accurate prediction of intron-exon donor boundaries for at least 7 (probably all 9) Dujonian 1st class introns. A computer-assisted search for homologies between mosaic genes transcripts and sequences complementary to tRNA1Lys reveals most of the control sequences previously determined as the sites of 1) cis-acting splicing blocks, 2) conserved RNA sequences and 3) conserved features of transcripts secondary structure. The sequences revealed in our study are as follows: sequences GGGGG and GGCTC in 2nd class introns and less convincing cases are R + E', E + P and IG in 1st class introns. The results obtained here together with the results of other authors suggest a series of (often weak) alternative RNA-RNA interactions needed to regulate and(or) provide a sufficiently error-free mechanism of splicing for the mosaic genes transcripts. A comparison was made of: 1) sequences near the sites of 2 different autocatalytic Tetrahymena pre-rRNA cleavage-ligation events (splicing and cyclization of the intron RNA) and 2) sequences of tRNA1Lys probably contacting donor boundaries of 1st class mitochondrial introns. The comparison reveals an obvious conservation of some basic structure regardless of the different chemical directions of the strands involved. The result suggests a possible catalytic function for the tRNA1Lys probably involved also in recognition and control processes of mitochondrial transcripts splicing.


Host interleukin-28B genetic variants versus viral kinetics in determining responses to standard-of-care for Asians with hepatitis C genotype 1.
Both interleukin-28B genetic variants and on-treatment virological responses are factors predictive of treatment outcome in hepatitis C virus genotype 1 (HCV-1) patients. We aimed to compare the clinical significance of the two factors. Rs8099917 genotype and on-treatment responses were determined in 182 HCV-1 patients with 48-week peginterferon/ribavirin. Comparing to patients with rs8099917 TG/GG genotype, those with TT genotype had significantly higher rapid virological response (RVR, 46.2% vs. 19.2%, P=0.01) and sustained virological response (SVR, 85.3% vs. 42.3%, P<0.001) rates. Logistic regression analysis revealed that the strongest factor predictive of a RVR was the carriage of rs8099917 TT genotype (odds ratio/95% confidence intervals [OR/CI]: 4.25/1.39-13.01). The most important factor predictive of an SVR was the attainment of a RVR (OR/CI: 57.22/6.23-525.37), followed by the carriage of rs8099917 TT genotype (OR/CI: 10.06/3.12-32.44). However, while on-treatment factors were taken into account, the cEVR was the most important determinant to an SVR (OR/CI:54.98/9.07-333.38), whereas the influence of rs8099917 genotype became non-significant in non-RVR patients. Rs8099917 TT genotype is significantly independently predictive of on-treatment virological responses, which were the major determinants of an SVR, in Asian HCV-1 patients.


Development of an online, patient-centred decision aid for patients with oropharyngeal cancer in the transoral robotic surgery era.
Radiotherapy (rt) has been the standard treatment for early oropharyngeal cancer, achieving excellent outcomes, but with significant toxicities. Transoral robotic surgery (tors) has emerged as a promising alternative. A decision aid (da) can help to establish patient treatment preferences. A da was developed and piloted in 40 healthy adult volunteers. Assuming equal oncologic outcomes of the treatments, participants indicated their preference. The treatment trade-off point was then established, and participant perceptions were elicited. More than 80% of participants initially selected tors for treatment, regardless of facilitator background. For all participants, the treatment trade-off point changed after an average 15% cure benefit. Treatment toxicities, duration, novelty, and perceptions all influenced treatment selection. All subjects valued the da. A da developed for early oropharyngeal cancer treatment holds promise in the era of shared decision-making. Assuming equal cure rates, tors was preferred over rt by healthy volunteers.


Clinical epidemiology and paraclinical findings in tuberculosis patients in north of Iran.
Mycobacterium tuberculosis (M.TB) causes a wide spectrum of clinical diseases. The prevalence of TB is different in various parts of Iran and throughout the world. The present study aimed to determine the clinical epidemiology and paraclinical findings of TB. A cross-sectional study was conducted from 2008 to 2013. Patient demographic, clinical, and radiologic characteristics, picked up from the TB patient's files, were collected using a standard questionnaire format. Data was entered and analyzed using the SPSS version 16 statistical software and P value < 0.05 was considered statistically significant. Out of 212 patients enrolled in this study 62% were male and the mean age was about 50 years old. 98.6% were Iranian, and 46.2% were rural. Prevalence of smear-positive TB was 66.4%. Prevalence of positive PPD was 50.7% with no significant difference between HIV-positive and -negative patients (P = 0.8). Prevalence of diabetes mellitus was 17%. 36% of the patients had history of smoking and about 29.3% were addicted to narcotics. Cough was the most common symptom (94.5%) and 84% had sputum. 15 cases (7%) had extrapulmonary TB. The mean time between the onset of symptoms and admission was 46.5 days. The delay for admission between urban and rural populations was not significantly different (P = 0.68); but for those who were in prison, the delay was significant (P = 0.02). About 46% of the patients had cavitary lesions in CXRs. Timely diagnosis of TB especially in prisoners by understanding its most important epidemiologic characteristics and clinical features can help to make an early treatment and prevent spread of mycobacteria and their complications.