Systematic Reviews and Meta-analyses.
The rising number of medical publications makes it difficult to keep up-to-date on scientific knowledge. In recent years, reviews in the form of narrative or systematic publications and meta-analyses have increased. These can only be interpreted and evaluated if the reader understands the techniques used. This review article describes the differences between narrative and systematic reviews, together with the characteristics of meta-analysis, and discusses their interpretation. The concept of systematic reviews and meta-analysis includes a systematic literature search and summary, together with an appraisal of the quality of the publications. Systematic reviews are often considered to be original studies due to their structure and ability to reduce bias.


Factors influencing the food choices of Irish children and adolescents: a qualitative investigation.
Food choices established during childhood and adolescence tend to persist into adulthood with consequences for long-term health. Yet, to date, relatively little research has examined factors that influence the food choices of children and adolescents from their perspectives. In this article, previous research is extended by examining developmental differences between children's and adolescents' perceptions of factors influencing their food choices. Focus group discussions were conducted with 29 young people from three age groups (9-10, 13-14 and 16-18 years). An inductive thematic analysis identified three key factors as influencing food choices. These factors included intra-individual factors: the link between food preferences and awareness of healthy eating; intra-familial factors: the role of the home food environment; and extra-familial factors: eating away from the home. Findings indicate that there were developmental differences between children's and adolescents' perceptions of factors influencing food choice. Among adolescents, parental control began to diminish and adolescents exercised increased autonomy over their food choices compared with children. To develop effective nutrition interventions, it is important to gather child and adolescent input regarding factors perceived as influencing their food choices.


Experimental methods of molecular matter-wave optics.
We describe the state of the art in preparing, manipulating and detecting coherent molecular matter. We focus on experimental methods for handling the quantum motion of compound systems from diatomic molecules to clusters or biomolecules.Molecular quantum optics offers many challenges and innovative prospects: already the combination of two atoms into one molecule takes several well-established methods from atomic physics, such as for instance laser cooling, to their limits. The enormous internal complexity that arises when hundreds or thousands of atoms are bound in a single organic molecule, cluster or nanocrystal provides a richness that can only be tackled by combining methods from atomic physics, chemistry, cluster physics, nanotechnology and the life sciences.We review various molecular beam sources and their suitability for matter-wave experiments. We discuss numerous molecular detection schemes and give an overview over diffraction and interference experiments that have already been performed with molecules or clusters.Applications of de Broglie studies with composite systems range from fundamental tests of physics up to quantum-enhanced metrology in physical chemistry, biophysics and the surface sciences.Nanoparticle quantum optics is a growing field, which will intrigue researchers still for many years to come. This review can, therefore, only be a snapshot of a very dynamical process.


Online extraction and single trial analysis of regions contributing to erroneous feedback detection.
Understanding how the brain processes errors is an essential and active field of neuroscience. Real time extraction and analysis of error signals provide an innovative method of assessing how individuals perceive ongoing interactions without recourse to overt behaviour. This area of research is critical in modern Brain-Computer Interface (BCI) design, but may also open fruitful perspectives in cognitive neuroscience research. In this context, we sought to determine whether we can extract discriminatory error-related activity in the source space, online, and on a trial by trial basis from electroencephalography data recorded during motor imagery. Using a data driven approach, based on interpretable inverse solution algorithms, we assessed the extent to which automatically extracted error-related activity was physiologically and functionally interpretable according to performance monitoring literature. The applicability of inverse solution based methods for automatically extracting error signals, in the presence of noise generated by motor imagery, was validated by simulation. Representative regions of interest, outlining the primary generators contributing to classification, were found to correspond closely to networks involved in error detection and performance monitoring. We observed discriminative activity in non-frontal areas, demonstrating that areas outside of the medial frontal cortex can contribute to the classification of error feedback activity.


Extractive microbial fermentation in cloud point system.
Extractive microbial fermentation of organic compounds in liquid-liquid two-phase systems is a potential strategy to overcome the limitations of microbial fermentation in an aqueous solution, such as low substrate solubility, substrate/product inhibition and product further degradation. A conventional aqueous-organic solvent two-phase system is inaccessible to extractive fermentation of a relatively high polar bioproduct as the confliction between the biocompatibility and the extraction ability of the corresponding organic solvent. An exploitation of cloud point system as a novel medium engineering method for extractive microbial fermentation is reviewed in present work. The relationship between phase separation of nonionic surfactant aqueous solution forming cloud point system and its corresponding biocompatibility to microorganisms, and the relationship between solubilization and bioavailability of organic compounds in a cloud point system are discussed. Paradigms of extractive microbial fermentation in cloud point system are highlighted with some cases in our lab. The downstream processing for nonionic surfactant recovery and product separation with microemulsion extraction is also presented.


The cAMP analog 8-Cl-cAMP inhibits growth and induces differentiation and apoptosis in retinoblastoma cells.
Retinoblastomas appear to be derived from a multipotential stem cell of the retina, due to alterations of the Rb1 gene. These tumors arise only within a discrete time frame during childhood, prior to terminal differentiation of the retinal precursor cells. Treatment of retinoblastoma cells with certain agents can induce a partial differentiation of cell types resembling those of the mature retina, such as rod and cone photoreceptors, glia, conventional neurons and pigment epithelia. We have tested the effects of 8-Cl-cAMP, a synthetic analog of cAMP which preferentially binds to and activates the RII subunit of protein kinase A on the Y-79 retinoblastoma cell line in vitro. Y-79 cells treated with 8-Cl-cAMP produced short, branching processes and showed a substantial increase in staining for neuron-specific enolase, a marker for conventional neuronal differentiation. In contrast, dibutyryl-cAMP gives a strong increase in the glial marker glial acidic fibrillary protein. Y-79 cell proliferation was strongly inhibited by 8-Cl-cAMP at concentrations as low as 5-25 microM. 8-Cl-cAMP significantly increased the rate of apoptosis of Y-79 cells in a dose-dependent manner. It also modulated expression of the RI regulatory subunit of intracellular cAMP-dependent protein kinase A, which is produced in abnormal quantities by Y-79 cells. A decrease in protein production was observed, with no clear effect on the RI subunit mRNA expression, suggesting that RI regulation occurs post-transcriptionally.


A brief review of the biochemistry and pharmacology of Eglin c, an elastase inhibitor.
Eglin c is a small protein inhibitor of human leucocyte elastase and cathepsin G which has potential as a therapeutic agent for the treatment of disease states associated with inflammation. In particular, Eglin c has been proven effective in experimental animal models of emphysema and shock. While initial studies indicate that Eglin c is virtually non-toxic and does not produce deleterious effects to the cardiovascular system, central nervous system or basic metabolism (unpublished results), an allergenic potential of the protein has not yet been ruled out.


Heterotypic Stressors Unmask Behavioral Influences of PMAT Deficiency in Mice.
Certain life stressors having enduring physiological and behavioral consequences, in part by eliciting dramatic signaling shifts in monoamine neurotransmitters. High monoamine levels can overwhelm selective transporters like the serotonin transporter. This is when polyspecific transporters like plasma membrane monoamine transporter (PMAT, Slc29a4) are hypothesized to contribute most to monoaminergic signaling regulation. Here, we employed two distinct counterbalanced stressors-fear conditioning and swim stress-in mice to systematically determine how reductions in PMAT function affect heterotypic stressor responsivity. We hypothesized that male heterozygotes would exhibit augmented stressor responses relative to female heterozygotes. Decreased PMAT function enhanced context fear expression, an effect unexpectedly obscured by a sham stress condition. Impaired cued fear extinction retention and enhanced context fear expression in males were conversely unmasked by a sham swim condition. Abrogated corticosterone levels in male heterozygotes that underwent swim stress after context fear conditioning did not map onto any measured behaviors. In sum, male heterozygous mouse fear behaviors proved malleable in response to preceding stressor or sham stress exposure. Combined, these data indicate that reduced male PMAT function elicits a form of stress-responsive plasticity. Future studies should assess how PMAT is differentially affected across sexes and identify downstream consequences of the stress-shifted corticosterone dynamics.


A novel SERCA inhibitor demonstrates synergy with classic SERCA inhibitors and targets multidrug-resistant AML.
Drug resistance exists as a major obstacle in the treatment of cancer, and drug molecules that retain effectiveness against resistant cancers are a high clinical priority. Ethyl 2-amino-6-(3,5-dimethoxyphenyl)-4-(2-ethoxy-2-oxoethyl)-4H-chromene-3-carboxylate (CXL017) was recently identified as a promising lead for the treatment of multidrug-resistant leukemia, which elicits its cytotoxic effect, in part, through inhibition of the sarco/endoplasmic reticulum Ca(2+)-ATPase (SERCA). Herein initial experiments with SERCA1a and CXL017 demonstrated no significant effect on calcium affinity, competed with ATP, and induced a dose-dependent decrease in ATPase activity. Among all CXLs tested, (-)-CXL017 exhibited the greatest SERCA inhibition with an IC50 = 13.5 ± 0.5 μM. Inhibitor combination studies were used to assess potential interactions between (-)-CXL017 and well-known SERCA inhibitors: thapsigargin, cyclopiazonic acid, and 2,5-di-tert-butylhydroquinone. Surprisingly, (-)-CXL017 exhibited marked synergy with each of the known SERCA inhibitors, whereas all combinations of the known inhibitors yielded additive effects, indicating that (-)-CXL017 may bind at a unique allosteric site. Treatment of parental (HL60) and multidrug-resistant (HL60/MX2) acute myeloid leukemia cells with the known SERCA inhibitors revealed that all of these inhibitors demonstrate selective cytotoxicity (7.7-400-fold) for the resistant cell line. Within the CXL series, a positive correlation exists between SERCA inhibition and cytotoxicity in HL60/MX2 but not HL60. (-)-CXL017 was also shown to enhance the cytotoxicity of thapsigargin in HL60/MX2 cells. Given the elevated SERCA levels and ER calcium content in HL60/MX2, SERCA likely plays a significant role in the collateral sensitivity of this multidrug-resistance cell line to CXL molecules as well as known SERCA inhibitors.


Inactivation of FAM20B causes cell fate changes in annulus fibrosus of mouse intervertebral disc and disc defects via the alterations of TGF-β and MAPK signaling pathways.
Intervertebral disc (IVD) disorder is often caused by the defect of annulus fibrosus (AF), especially that of the outer AF. Studies about the mechanisms governing the development of the outer AF are needed for a better understanding of pathogenesis of IVD defects. Glycosaminoglycans (GAGs) are essential components of extracellular matrix (ECM) in AF. FAM20B is a newly identified xylose kinase that catalyzes the biosynthesis of GAGs. In this study, we created Fam20B conditional knockout (cKO) mice in which FAM20B was inactivated in type I collagen-expressing cells, the main type of cells in the outer AF of IVD. The cKO mice showed severe spine deformity and remarkable IVD defects associated with AF malformation. The AF of cKO mice had a lower level of chondroitin sulfate and heparan sulfate, and the outer AF cells lost their normal fibroblast-like morphology and acquired chondrocyte phenotypes, expressing a higher level of Sox 9 and type II collagen along with a reduced level of type I collagen. The level of phospho-Smad 2 and phospho-Smad 3, and that of scleraxis, a downstream target molecule of canonical TGF-β signaling pathway were significantly lower in the AF of cKO mice. The AF in cKO mice also manifested altered levels in the molecules associated with the activations of MAPK pathway; the changes included the increase of phospho-P38 and phospho-ERK and a decrease of phospho-JNK. These results indicate that FAM20B plays an essential role in the development of AF by regulating the TGF-β signaling and MAPK signaling pathways.


Effects of (+/-) 3,4-methylenedioxymethamphetamine (MDMA) on sleep and circadian rhythms.
Abuse of stimulant drugs invariably leads to a disruption in sleep-wake patterns by virtue of the arousing and sleep-preventing effects of these drugs. Certain stimulants, such as 3,4-methylenedioxymethamphetamine (MDMA), may also have the potential to produce persistent alterations in circadian regulation and sleep because they can be neurotoxic toward brain monoaminergic neurons involved in normal sleep regulation. In particular, MDMA has been found to damage brain serotonin (5-HT) neurons in a variety of animal species, including nonhuman primates, with growing evidence that humans are also susceptible to MDMA-induced brain 5-HT neurotoxicity. 5-HT is an important modulator of sleep and circadian rhythms and, therefore, individuals who sustain MDMA-induced 5-HT neurotoxicity may be at risk for developing chronic abnormalities in sleep and circadian patterns. In turn, such abnormalities could play a significant role in other alterations reported in abstinent in MDMA users (e.g., memory disturbance). This paper will review preclinical and clinical studies that have explored the effects of prior MDMA exposure on sleep, circadian activity, and the circadian pacemaker, and will highlight current gaps in knowledge and suggest areas for future research.


Biosynthesis of zinc oxide nanoparticles using anjbar (root of Persicaria bistorta) extract and their cytotoxic effects on human breast cancer cell line (MCF-7).
Biosynthesis of nanoparticles (NPs) using biomass is now one of the best methods for synthesising NPs due to their nontoxic and biocompatibility. Plants are the best choice among all biomass to synthesise large-scale NPs. The objectives of this study were to synthesise zinc oxide nanoparticles (ZnO-NPs) using Anjbar (root of Persicaria bistorta) [An/ZnO-NPs] and investigate the cytotoxic and anti-oxidant effects. For this purpose, the An/ZnO-NPs were synthesised by using Bistort extract and characterised using UV-Visible spectroscopy, transmission electron microscope, field emission scanning electron microscope, x-ray diffraction and Fourier-transform infrared spectroscopy. The cytotoxic effects of the An/ZnO-NPs on MCF-7 cells were followed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays at 24, 48, and 72 h. Nuclear morphology changed and apoptosis in cells was investigated using acridine orange/propodium iodide (AO/PI) staining and flow cytometry analysis. The pure biosynthesised ZnO-NPs were spherical in shape and particles sizes ranged from 1 to 50 nm. Treated MCF-7 cells with different concentrations of ZnO-NPs inhibited cell viability in a time- and dose-dependent manner with IC50 about 32 μg/ml after 48 h of incubation. In flow cytometry analysis the sub-G1 population, which indicated apoptotic cells, increased from 12.6% at 0 μg/ml (control) to 92.8% at 60 μg/ml, 48 h after exposure. AO/PI staining showed that the treated cells displayed morphologic evidence of apoptosis, compared to untreated groups.


How we do it: a combined method of traditional curette and power-assisted endoscopic adenoidectomy.
After conventional curette adenoidectomy, a significant mass of residual adenoid tissue is observed in about 50% of the cases. The combined approach of conventional curette and endoscopic adenoidectomy with microdebrider assures a complete and accurate removal of the mass. It is an effective and safe method without significantly prolonging the operative time. To describe the surgical technique of conventional curette adenoidectomy completed with the microdebrider under endoscopic vision in case of residual adenoid tissue. This was a retrospective review of 143 consecutive patients (age range 2-16 years, mean 6.2) who underwent endoscopic adenoidectomy for adenoid hypertrophy at an academic hospital-based institution. A classic transoral curette adenoidectomy was performed initially. Then a rigid fibre-optic endoscope was used transnasally to assess the completeness of surgery. Residual adenoid tissue was removed using a microdebrider under endoscopic vision. After conventional curette adenoidectomy 70 children (48.9%) underwent residual adenoid tissue transnasal removal. Mean curette adenoidectomy time with endoscopic visualization of the nasopharynx was 9.1 min and, in the case of completion with the microdebrider, 14.6 min. No significant intraoperative bleeding, postoperative haemorrhage or other complications have occurred. After a mean follow-up of 24.4 months, no patients have been readmitted for symptoms of adenoid regrowth.


Effects of ECG gating and postprocessing techniques on 3D MDCT of the bronchial tree.
Our goal was to determine the impact of ECG gating and different postprocessing techniques on 3D imaging of the bronchial tree. SUBJECTS AND METHODS. Retrospective ECG-gated MDCT and non-ECG-gated MDCT of the chest were performed in 25 patients. ECG-gated MDCT data were reconstructed mid diastole using a fixed interval of -400 msec in 25 patients and then additionally at -200, -300, and -500 msec in 10 of those patients. Shaded surface display and volume rendering of the bronchial tree combined with virtual bronchoscopy were performed using all data sets. The extent of bronchial tree visualization in shaded surface display-virtual bronchoscopy and volume rendering-virtual bronchoscopy and the presence of artifacts in volume-rendered images were scored by three blinded reviewers. The effective radiation doses of the ECG-gated and nongated acquisitions were compared. The summary scores of all bronchial segments for gated shaded surface display-virtual bronchoscopy and gated volume rendering-virtual bronchoscopy did not differ significantly. The summary scores for nongated shaded surface display-virtual bronchoscopy and nongated volume rendering-virtual bronchoscopy were not significantly different. Non-gated acquisition yielded significantly better visualization of the bronchial tree for both post-processing techniques, regardless of the time interval used for reconstruction of the ECG-gated series. Artifact scores in volume-rendered images were significantly higher for ECG-gated MDCT compared with nongated MDCT. Effective radiation dose was significantly higher for the ECG-gated acquisition. Given the advantage of volume rendering for representing the entire data set and given the lower radiation dose and better 3D image quality of nongated acquisition, volume rendering performed on nongated MDCT data is the method of choice for 3D visualization of the bronchial tree.


[Comparison of the protein spectra of typical and atypical mycobacteria using the disc electrophoresis method in polyacrylamide gel].
Protein spectra of 19 strains of typical and atypical mycobacteria were studied. The advantage of protein extracts obtained from the living cells destroyed by press was shown in experiments of fractionation in polyacrylamide gel. On the basis of similarity indices of the protein spectra a conditioned scheme reflecting the extent of similarity of the cultures under study was drawn. there were revealed protein fractions common for the majority of the cultures studied; It was impossible to isolate the fractions strictly specific of individual groups.


Tumor-specific recognition of human myeloma cells by idiotype-induced CD8(+) T cells.
Immunoglobulin secreted by myeloma cells contains a unique antigenic determinant (idiotype [Id]) that may serve as a tumor-specific antigen. Although Id-protein-specific T-cell responses have been reported in patients with myeloma, it is not known whether primary myeloma tumor cells can present naturally processed Id peptides on their surface as a target. We immunized 2 healthy human stem-cell donors with Id proteins from their recipients. T cells from the immunized donors released high levels of T-helper 1-type cytokines in response to stimulation with myeloma cells from their recipients. The T-cell-mediated cytokine response to tumor cells was blocked by a major histocompatibility complex (MHC) class I monoclonal antibody, whereas the response to soluble Id protein was dependent on MHC class II. To investigate whether Id-specific CD8(+) T cells can recognize and kill autologous myeloma cells, we generated T cells from peripheral blood mononuclear cells from a third patient with myeloma by means of in vitro stimulation with autologous dendritic cells pulsed with Id protein. Tumor-specific lysis of myeloma cells was demonstrated by the lack of killing of autologous nonmalignant B cells or natural killer-sensitive K562 cells. Lysis of autologous myeloma targets was restricted by MHC class I molecules. These data represent the first report of class I-restricted T-cell recognition of fresh autologous myeloma targets and formally demonstrate that human myeloma cells can serve as targets of an Id-specific T-cell response. (Blood. 2000;96:2828-2833)


Antineuronal antibodies against neurotransmitter receptors and synaptic proteins in schizophrenia: current knowledge and clinical implications.
When Eugen Bleuler coined the term 'schizophrenia' he believed that various causes of illness may underlie this disease. Currently, neurodevelopmental abnormalities and consecutive impairments in dopaminergic and glutamatergic neurotransmission are considered as major causes of schizophrenia. However, there are various indications for involvement of immune processes, at least in subgroups of patients. Circulating antineuronal antibodies provide a promising link between the well-described disturbances in neurotransmission and the immune hypothesis of schizophrenia. This review summarizes important studies that have examined the role of glutamate, dopamine, acetylcholine and serotonin receptor autoantibodies, and other antineuronal antibodies against synaptic proteins in the serum of patients diagnosed with schizophrenia. Currently, it is not known whether the presence of antineuronal antibodies in blood should be considered as a causal or disease-modulating factor in schizophrenia. Due to emerging evidence regarding the important role of the blood-brain barrier, combined testing of serum and cerebrospinal fluid is likely to be more appropriate to answer this question than pure serum analyses. We suggest implementation of such testing in first-onset and treatment-resistant patients as part of the diagnostic process. In addition, future clinical trials should evaluate if immunotherapy (e.g. cortisone pulse therapy, intravenous immunoglobulins, plasmapheresis, rituximab, or cyclophosphamide) is helpful in cases with a neuroinflammatory component.


Molecular identification of Knops blood group polymorphisms found in long homologous region D of complement receptor 1.
Complement receptor 1 (CR1) has been implicated in rosetting of uninfected red blood cells to Plasmodium falciparum-infected cells, and rosette formation is associated with severe malaria. The Knops blood group (KN) is located on CR1 and some of these antigens, ie, McCoy (McC) and Swain-Langley (Sl(a)), show marked frequency differences between Caucasians and Africans. Thus, defining the molecular basis of these antigens may provide new insight into the mechanisms of P falciparum malaria. Monoclonal antibody epitope mapping and serologic inhibition studies using CR1 deletion constructs localized McC and Sl(a) to long homologous repeat D of CR1. Direct DNA sequencing of selected donors identified several single nucleotide polymorphisms in exon 29 coding for complement control protein modules 24 and 25. Two of these appeared to be blood group specific: McC associated with K1590E and Sl(a) with R1601G. These associations were confirmed by inhibition studies using allele-specific mutants. A sequence-specific oligonucleotide probe hybridization assay was developed to genotype several African populations and perform family inheritance studies. Concordance between the 1590 mutation and McC was 94%; that between Sl(a) and 1601 was 88%. All but 2 samples exhibiting discrepancies between the genotype and phenotype were found to be due to low red cell CR1 copy numbers, low or absent expression of some alleles, or heterozygosity combined with low normal levels of CR1. These data further explain the variability observed in previous serologic studies of CR1 and show that DNA and protein-based genetic studies will be needed to clarify the role of the KN antigens in malaria.


Cytotoxicity and oxidative stress induced by different metallic nanoparticles on human kidney cells.
Some manufactured nanoparticles are metal-based and have a wide variety of applications in electronic, engineering and medicine. Until now, many studies have described the potential toxicity of NPs on pulmonary target, while little attention has been paid to kidney which is considered to be a secondary target organ. The objective of this study, on human renal culture cells, was to assess the toxicity profile of metallic nanoparticles (TiO2, ZnO and CdS) usable in industrial production. Comparative studies were conducted, to identify whether particle properties impact cytotoxicity by altering the intracellular oxidative status. Nanoparticles were first characterized by size, surface charge, dispersion and solubility. Cytotoxicity of NPs was then evaluated in IP15 (glomerular mesangial) and HK-2 (epithelial proximal) cell lines. ZnO and CdS NPs significantly increased the cell mortality, in a dose-dependent manner. Cytotoxic effects were correlated with the physicochemical properties of NPs tested and the cell type used. Analysis of reactive oxygen species and intracellular levels of reduced and oxidized glutathione revealed that particles induced stress according to their composition, size and solubility. Protein involved in oxidative stress such as NF-κb was activated with ZnO and CdS nanoparticles. Such effects were not observed with TiO2 nanoparticles. On glomerular and tubular human renal cells, ZnO and CdS nanoparticles exerted cytotoxic effects that were correlated with metal composition, particle scale and metal solubility. ROS production and oxidative stress induction clearly indicated their nephrotoxic potential.


Adipose tissue-derived mesenchymal stem cells and hepatic differentiation: old concepts and future perspectives.
Mesenchymal stem cells (MSCs) are multipotent cells, able to differentiate into elements of the mesodermal lineage. Bone marrow and adipose tissue represent the main sources for MSC isolation. In the last decade, several studies have reported the plasticity of MSCs toward a hepatocyte-like phenotype. The use of MSCs to generate hepatocyte-like cells holds great promises to overcome the scarcity of available organs for transplantation. However, little is known about the molecular pathways involved in lineage cross-differentiation and several issues remain to be answered before MSC application in clinical settings. Aim of this review is to critically analyze the possible sources of MSCs suitable for liver repopulation and the molecular mechanisms underlying MSC hepatic differentiation.


gamma-tocopherol and its major metabolite, in contrast to alpha-tocopherol, inhibit cyclooxygenase activity in macrophages and epithelial cells.
Cyclooxygenase-2 (COX-2)-catalyzed synthesis of prostaglandin E(2) (PGE(2)) plays a key role in inflammation and its associated diseases, such as cancer and vascular heart disease. Here we report that gamma-tocopherol (gammaT) reduced PGE(2) synthesis in both lipopolysaccharide (LPS)-stimulated RAW264.7 macrophages and IL-1beta-treated A549 human epithelial cells with an apparent IC(50) of 7.5 and 4 microM, respectively. The major metabolite of dietary gammaT, 2,7,8-trimethyl-2-(beta-carboxyethyl)-6-hydroxychroman (gamma-CEHC), also exhibited an inhibitory effect, with an IC(50) of approximately 30 microM in these cells. In contrast, alpha-tocopherol at 50 microM slightly reduced (25%) PGE(2) formation in macrophages, but had no effect in epithelial cells. The inhibitory effects of gammaT and gamma-CEHC stemmed from their inhibition of COX-2 activity, rather than affecting protein expression or substrate availability, and appeared to be independent of antioxidant activity. gamma-CEHC also inhibited PGE(2) synthesis when exposed for 1 h to COX-2-preinduced cells followed by the addition of arachidonic acid (AA), whereas under similar conditions, gammaT required an 8- to 24-h incubation period to cause the inhibition. The inhibitory potency of gammaT and gamma-CEHC was diminished by an increase in AA concentration, suggesting that they might compete with AA at the active site of COX-2. We also observed a moderate reduction of nitrite accumulation and suppression of inducible nitric oxide synthase expression by gammaT in lipopolysaccharide-treated macrophages. These findings indicate that gammaT and its major metabolite possess anti-inflammatory activity and that gammaT at physiological concentrations may be important in human disease prevention.


Development and validation of open-source deep neural networks for comprehensive chest x-ray reading: a retrospective, multicentre study.
Artificial intelligence (AI) systems for automated chest x-ray interpretation hold promise for standardising reporting and reducing delays in health systems with shortages of trained radiologists. Yet, there are few freely accessible AI systems trained on large datasets for practitioners to use with their own data with a view to accelerating clinical deployment of AI systems in radiology. We aimed to contribute an AI system for comprehensive chest x-ray abnormality detection. In this retrospective cohort study, we developed open-source neural networks, X-Raydar and X-Raydar-NLP, for classifying common chest x-ray findings from images and their free-text reports. Our networks were developed using data from six UK hospitals from three National Health Service (NHS) Trusts (University Hospitals Coventry and Warwickshire NHS Trust, University Hospitals Birmingham NHS Foundation Trust, and University Hospitals Leicester NHS Trust) collectively contributing 2 513 546 chest x-ray studies taken from a 13-year period (2006-19), which yielded 1 940 508 usable free-text radiological reports written by the contemporary assessing radiologist (collectively referred to as the "historic reporters") and 1 896 034 frontal images. Chest x-rays were labelled using a taxonomy of 37 findings by a custom-trained natural language processing (NLP) algorithm, X-Raydar-NLP, from the original free-text reports. X-Raydar-NLP was trained on 23 230 manually annotated reports and tested on 4551 reports from all hospitals. 1 694 921 labelled images from the training set and 89 238 from the validation set were then used to train a multi-label image classifier. Our algorithms were evaluated on three retrospective datasets: a set of exams sampled randomly from the full NHS dataset reported during clinical practice and annotated using NLP (n=103 328); a consensus set sampled from all six hospitals annotated by three expert radiologists (two independent annotators for each image and a third consultant to facilitate disagreement resolution) under research conditions (n=1427); and an independent dataset, MIMIC-CXR, consisting of NLP-annotated exams (n=252 374). X-Raydar achieved a mean AUC of 0·919 (SD 0·039) on the auto-labelled set, 0·864 (0·102) on the consensus set, and 0·842 (0·074) on the MIMIC-CXR test, demonstrating similar performance to the historic clinical radiologist reporters, as assessed on the consensus set, for multiple clinically important findings, including pneumothorax, parenchymal opacification, and parenchymal mass or nodules. On the consensus set, X-Raydar outperformed historical reporter balanced accuracy with significance on 27 of 37 findings, was non-inferior on nine, and inferior on one finding, resulting in an average improvement of 13·3% (SD 13·1) to 0·763 (0·110), including a mean 5·6% (13·2) improvement in critical findings to 0·826 (0·119). Our study shows that automated classification of chest x-rays under a comprehensive taxonomy can achieve performance levels similar to those of historical reporters and exhibit robust generalisation to external data. The open-sourced neural networks can serve as foundation models for further research and are freely available to the research community. Wellcome Trust.


Projecting Month of Birth for At-Risk Infants after Zika Virus Disease Outbreaks.
The marked increase in infants born with microcephaly in Brazil after a 2015 outbreak of Zika virus (Zika virus) disease suggests an association between maternal Zika virus infection and congenital microcephaly. To project the timing of delivery of infants born to mothers infected during early pregnancy in 1 city in Bahia State, Brazil, we incorporated data on reported Zika virus disease cases and microcephaly cases into a graphical schematic of weekly birth cohorts. We projected that these births would occur through February 2016. Applying similar projections to a hypothetical location at which Zika virus transmission started in November, we projected that full-term infants at risk for Zika virus infection would be born during April-September 2016. We also developed a modifiable spreadsheet tool that public health officials and researchers can use for their countries to plan for deliveries of infants to women who were infected with Zika virus during different pregnancy trimesters.


Support vector machines in DSC-based glioma imaging: suggestions for optimal characterization.
Dynamic susceptibility contrast magnetic resonance perfusion imaging (DSC-MRI) is a useful method to characterize gliomas. Recently, support vector machines (SVMs) have been introduced as means to prospectively characterize new patients based on information from previous patients. Based on features derived from automatically segmented tumor volumes from 101 DSC-MR examinations, four different SVM models were compared. All SVM models achieved high prediction accuracies (>82%) after rebalancing the training data sets to equal amounts of samples per class. Best discrimination was obtained using a SVM model with a radial basis function kernel. A correct prediction of low-grade glioma was obtained at 83% (true positive rate) and for high-grade glioma at 91% (true negative rate) on the independent test data set. In conclusion, the combination of automated tumor segmentation followed by SVM classification is feasible. Thereby, a powerful tool is available to characterize glioma presurgically in patients.


A Preclinical Validation of Iron Oxide Nanoparticles for Treatment of Perianal Fistulizing Crohn's Disease.
Fistulizing anoperineal lesions are severe complications of Crohn's disease (CD) that affect quality of life with a long-term risk of anal sphincter destruction, incontinence, permanent stoma, and anal cancer. Despite several surgical procedures, they relapse in about two-thirds of patients, mandating innovative treatments. Ultrasmall particles of iron oxide (USPIO) have been described to achieve in vivo rapid healing of deep wounds in the skin and liver of rats thanks to their nanobridging capability that could be adapted to fistula treatment. Our main purpose was to highlight preclinical data with USPIO for the treatment of perianal fistulizing CD. Twenty male Sprague Dawley rats with severe 2,4,6-trinitrobenzenesulfonic acid solution (TNBS)-induced proctitis were operated to generate two perianal fistulas per rat. At day 35, two inflammatory fistulas were obtained per rat and perineal magnetic resonance imaging (MRI) was performed. After a baseline MRI, a fistula tract was randomly drawn and topically treated either with saline or with USPIO for 1 min (n = 17 for each). The rats underwent a perineal MRI on postoperative days (POD) 1, 4, and 7 and were sacrificed for pathological examination. The primary outcome was the filling or closure of the fistula tract, including the external or internal openings. USPIO treatment allowed the closure and/or filling of all the treated fistulas from its application until POD 7 in comparison with the control fistulas (23%). The treatment with USPIO was safe, permanently closed the fistula along its entire length, including internal and external orifices, and paved new avenues for the treatment of perianal fistulizing Crohn's disease.


[Hand port-assisted laparoscopic surgery].
The dramatic benefits of laparoscopic cholecystectomy or appendectomy for patients encouraged surgeons to use minimal access operative technique in the treatment of other more complex surgical procedures [2, 3]. The authors report on their experiences with 14 patients operated by hand-port assisted laparoscopic technique for benign colorectal diseases. The mortality rate was zero. The operation times compared to the laparoscopically operated previous 14 patients with equal diagnosis did not differ significantly (hand-port: 124-186 min; lap.: 121-176 min). The begin of bowel movements and the postoperative hospital stay were comparable to those of laparoscopic surgery. There were no major complications. No conversion to an open procedure was necessary. As the hand-port device allows the surgeon to insert his (usually non-dominant) hand into the abdominal cavity during the procedure, the hand-port device seems to combine the laparoscopic benefits with the advantages of a conventional open approach (manual exploration, blunt dissection, control of hemostasis) without loss of pneumoperitoneum [10]. Even though hand-assisted operations have not gained widespread acceptance, they recently demonstrated their value especially in more complex laparoscopic procedures like splenic and gastric resections, nephrectomy and colorectal surgery [6, 10, 18, 21, 23-24]. The regaining of tactile sensation which is an essential surgical tool may encourage less experienced colleagues to perform more complex operations. The authors suggest that the hand-port device could be a useful tool in the armentarium for colorectal surgery. Further randomized trials are needed to evaluate the benefits of this technique.


The association between parental smoking and the diet quality of low-income children.
To assess the association between parental smoking and the diet quality of children residing in low-income housholds in the United States. MTHODS: Data from 515 low-income children (less than or equal to 185% of the poverty line), ages 2 to 17, who participated in the 1989 and 1990 United States Department of Agriculture Continuing Survey of Food Intakes of Individuals were examined. Diet quality was assessed by examining the average daily amount of nutrients consumed per 1000 kcal for protein, fiber, and 14 essential vitamins and minerals as well as total energy, percent of energy from total fat and saturated fat, and cholesterol and sodium intakes using the 3-day average of one 24-hour recall and 2 days of diet records. Parental smoking was categorized as four levels (nonsmoker; 1 to 10, 11 to 20, and more than 20) on the basis of the average number of cigarettes smoked per day by the sample child's parents. Analysis of covariance examined differences in the children's nutrient intake among the four smoking categories while controling for race, mother's age and occupation, child age, and sex. Low-income children with parents who smoked (n = 235) were more likely to be white (P <.001), had younger mothers (P <.05) and were more likely to have mothers employed in blue-collar occupations (P <.001) than children whose parents were nonsmokers (N = 280). Children whose parents smoked more than 20 cigarettes per day had a higher level of energy from saturated fat, and children whose parents smoked 11 to 20 cigarettes per day had the highest cholesterol intakes in comparison with the rest of the sample. Parental smoking was also related to total fiber intake per 1000 kcal, with children of smokers having lower fiber intakes than children of nonsmokers. On average, low-income children of smokers had a poorer diet quality than low-income children of nonsmokers, thus increasing their future risk of chronic disease.


Functional and metabolic properties of polymorphonuclear leucocytes. II. The influence of a lipopolysaccharide endotoxin.
The effects of a purified bacterial lipopolysaccharide endotoxin on homogenous populations of rabbit polymorphonuclear leucocytes have been studied in vitro under defined conditions. Employing a 500-fold range of concentration (0.1 to 50.0 microg./ml.), it was shown that endotoxin enhanced the rate at which staphylococci were killed by leucocytes. The mechanism underlying the increased killing was found to be a direct stimulation of the phagocytic activity of the leucocyte and not mediated by the release of bactericidins or opsonins from the treated cells. In the presence of 10 per cent serum all concentrations of endotoxin enhanced phagocytosis, whereas at lower serum concentrations, the higher doses of lipopolysaccharide inhibited the phagocytic activity of the cells. Similar concentrations of endotoxin were capable of increasing the utilization of glucose and the production of lactic acid. Endotoxin treated leucocytes exhibited no change in oxygen consumption, and only a slight depression in glycogen synthesis. It appeared that endotoxin could interact and alter the functional and metabolic properties of leucocytes in the absence of serum. The demonstration of enhanced phagocytic activity of endotoxin-treated cells was dependent upon the particular opsonic requirements for the organism under study.


Exercise echocardiographic comparison of pulmonary autograft and aortic homograft replacements for aortic valve disease in adults.
The pulmonary autograft, or Ross procedure, has theoretical hemodynamic benefits over other aortic valve replacements. The hemodynamic performance of the pulmonary autograft and pulmonary homograft components of this procedure have not been well defined. Twenty patients with pulmonary autograft replacement of the aortic valve and six with aortic homografts underwent exercise echocardiography with assessment of exercise duration, left ventricular dimensions, mass, and function. Hemodynamics at rest and maximal exercise, including Doppler gradients and effective orifice area (EOA), were measured across the pulmonary autograft and aortic homograft valves. Doppler gradients across the pulmonary homograft valves were compared to native pulmonary valve gradients at rest and maximal exercise. Both groups of patients had excellent self-reported and measured exercise capacity. In comparison to the aortic homograft, the pulmonary autograft had lower peak Doppler gradients across the neoaortic valve at rest (5 +/- 2 versus 11 +/- 4 mmHg; p = 0.027) and maximal exercise (10 +/- 5 versus 15 +/- 5 mmHg; p = 0.003) and larger indexed EOA. However, the Ross procedure patients had higher gradients across the pulmonary homograft both at rest (14 +/- 10 versus 3 +/- 1 mmHg; p < 0.001) and maximal exercise (25 +/- 22 versus 5 +/- 4 mmHg; p = 0.004). Two patients in the Ross procedure group had significant pulmonary homograft stenosis in short- or mid-term follow up. In comparison to aortic homograft replacement of the aortic valve, pulmonary autograft replacement has superior hemodynamics at rest and during exercise. However, the pulmonary homograft replacement may develop hemodynamically significant stenosis after the Ross procedure.


Comparing different estimates of cochlear compression in listeners with normal and impaired hearing.
A loss of cochlear compression may underlie many of the difficulties experienced by hearing-impaired listeners. Two behavioral forward-masking paradigms that have been used to estimate the magnitude of cochlear compression are growth of masking (GOM) and temporal masking (TM). The aim of this study was to determine whether these two measures produce within-subjects results that are consistent across a range of signal frequencies and, if so, to compare them in terms of reliability or efficiency. GOM and TM functions were measured in a group of five normal-hearing and five hearing-impaired listeners at signal frequencies of 1000, 2000, and 4000 Hz. Compression values were derived from the masking data and confidence intervals were constructed around these estimates. Both measures produced comparable estimates of compression, but both measures have distinct advantages and disadvantages, so that the more appropriate measure depends on factors such as the frequency region of interest and the degree of hearing loss. Because of the long testing times needed, neither measure is suitable for clinical use in its current form.


Vasculitis associated with infectious agents.
The HIV infection epidemic and, with it, the resurgence and emergence of old and new opportunistic microbes, have sparked off a renewed interest in vasculitis associated with infectious agents. Current investigations focus on identification of "newer" infectious agents, unusual clinical manifestations, and methodologic refinement in diagnosis and treatment. This article briefly reviews the background of infection-associated vasculitis and highlights some of the newer and unusual aspects of vasculitis known to be associated with nonviral and viral infections.


Conventional standard-sized bra with prosthesis or patient's bra with customized hand-knitted external prosthesis after mastectomy: Mixed-methods evaluation of patients' preferences.
In breast cancer patients undergoing mastectomy without reconstruction, an external breast prosthesis could aid patients' recovery, improve body image and confidence by helping to regain a symmetrical chest appearance when dressed. However, external breast prosthesis preferences among Asian breast cancer patients were not widely studied. We aimed to compare patients' experience with the conventional commercially manufactured standard-sized (small, medium, large, extra-large) bra and prosthesis versus customized hand-knitted external breast prosthesis with patient's bra after unilateral mastectomy at a tertiary hospital. This is the first such study in Asian women, to our knowledge. In this prospective study, participants used the conventional bra-prosthesis followed by the customized one consecutively, each for at least 3 months before they were administered an identical questionnaire at 3 and 6 months respectively. The questionnaire assessed the patients' experience with the prosthesis on the aspects of comfort, body image and satisfaction etc. Patients were also invited for in-depth interviews. Of 155 eligible patients, 148 patients participated with a response rate of 95.5%. 99 (67%) participants preferred the customized prosthesis, while 38 (25.7%) did not. 11 (7.4%) participants were undecided. Seventeen participants underwent in-depth interviews until data saturation on major qualitative themes was achieved. More patients experienced excessive sweating (p < 0.0001), greater discomfort (p = 0.0195) and higher rates of prosthesis dislodgement (p = 0.0269) with the conventional bra-prosthesis. Customized external breast prostheses could be an alternative to the conventional ones for breast cancer patients with mastectomy, with additional benefits of less sweating, more comfort and less dislodgement perceived.


Clinical case series: treatment of PTSD with transcendental meditation in active duty military personnel.
Active duty U.S. Army Service Members previously diagnosed with post-traumatic stress disorder (PTSD) were selected from review of patient records in the Traumatic Brain Injury Clinic at the Department of Defense Eisenhower Army Medical Center at Fort Gordon in Augusta, Georgia. Patients agreed to practice the Transcendental Meditation (TM) technique for 20 minutes twice a day for the duration of a 2-month follow-up period. Three cases are presented with results that show the feasibility of providing TM training to active duty soldiers with PTSD in a Department of Defense medical facility. Further investigation is suggested to determine if a TM program could be used as an adjunct for treatment of PTSD. Impact of this report is expected to expand the complementary and alternative evidence base for clinical care of PTSD.


Naturally occurring insulin autoantibodies in neonates of normal pregnancies and their relationship to insulinemia and birth weight.
The objectives of this study were to determine whether insulin autoantibodies are present in umbilical cord blood from normal pregnancies, determine whether cord blood insulin autoantibody levels correlate with respective maternal levels at delivery, determine whether cord blood insulin autoantibody levels are related to cord blood or maternal insulin levels, and to determine what relationship neonatal birth weight has with either cord blood insulin autoantibody and insulin levels or maternal insulin autoantibody and insulin levels. Paired umbilical cord and maternal serum samples were taken from 70 normal subjects at delivery. Measurements of serum insulin autoantibody (competitive charcoal radiobinding assay) and insulin (radioimmune inhibition assay) levels were performed. Multiple linear regression analysis and paired t tests were used for data analyses. Neonatal insulin autoantibody levels (120 nU/ml) were more than two times higher than maternal levels (49 nU/ml) (p < 0.001). No correlation was observed between neonatal and maternal insulin autoantibody levels (r = 0.14, p = 0.25). A positive correlation of both neonatal and maternal insulin with birth weight was observed (r = 0.28, p < 0.02; and r = 0.36, p < 0.01, respectively). These results suggest that the insulin autoantibody levels in fetal cord blood are not related to maternal levels in normal uncomplicated pregnancies. In addition, insulin levels in both maternal and neonatal circulations were positively correlated with increased birth weight in the normal pregnancies studied.


Sex Disparities in Ischemic Stroke Care: FL-PR CReSD Study (Florida-Puerto Rico Collaboration to Reduce Stroke Disparities).
Sex-specific disparities in stroke care including thrombolytic therapy and early hospital admission are reported. In a large registry of Florida and Puerto Rico hospitals participating in the Get With The Guidelines-Stroke program, we sought to determine sex-specific differences in ischemic stroke performance metrics and overall thrombolytic treatment. Around 51 317 (49% women) patients were included from 73 sites from 2010 to 2014. Multivariable logistic regression with generalized estimating equations evaluated sex-specific differences in the prespecified Get With The Guidelines-Stroke metrics for defect-free care in ischemic stroke, adjusting for age, race-ethnicity, insurance status, hospital characteristics, individual risk factors, and the presenting stroke severity. As compared with men, women were older (73±15 versus 69±14 years; P<0.0001), more hypertensive (67% versus 63%, P<0.0001), and had more atrial fibrillation (19% versus 16%; P<0.0001). Defect-free care was slightly lower in women than in men (odds ratio, 0.96; 95% confidence interval, 0.93-1.00). Temporal trends in defect-free care improved substantially and similarly for men and women, with a 29% absolute improvement in women (P<0.0001) and 28% in men (P<0.0001), with P value of 0.13 for time-by-sex interaction. Women were less likely to receive thrombolysis (odds ratio, 0.92; 95% confidence interval, 0.86-0.99; P=0.02) and less likely to have a door-to-needle time <1 hour (odds ratio, 0.83; 95% confidence interval, 0.71-0.97; P=0.02) as compared with men. Women received comparable stroke care to men in this registry as measured by prespecified Get With The Guidelines metrics. However, women less likely received thrombolysis and had door-to-needle time <1 hour, an observation that calls for the implementation of interventions to reduce sex disparity in these measures.


Ebf1 or Pax5 haploinsufficiency synergizes with STAT5 activation to initiate acute lymphoblastic leukemia.
As STAT5 is critical for the differentiation, proliferation, and survival of progenitor B cells, this transcription factor may play a role in acute lymphoblastic leukemia (ALL). Here, we show increased expression of activated signal transducer and activator of transcription 5 (STAT5), which is correlated with poor prognosis, in ALL patient cells. Mutations in EBF1 and PAX5, genes critical for B cell development have also been identified in human ALL. To determine whether mutations in Ebf1 or Pax5 synergize with STAT5 activation to induce ALL, we crossed mice expressing a constitutively active form of STAT5 (Stat5b-CA) with mice heterozygous for Ebf1 or Pax5. Haploinsufficiency of either Pax5 or Ebf1 synergized with Stat5b-CA to rapidly induce ALL in 100% of the mice. The leukemic cells displayed reduced expression of both Pax5 and Ebf1, but this had little effect on most EBF1 or PAX5 target genes. Only a subset of target genes was deregulated; this subset included a large percentage of potential tumor suppressor genes and oncogenes. Further, most of these genes appear to be jointly regulated by both EBF1 and PAX5. Our findings suggest a model whereby small perturbations in a self-reinforcing network of transcription factors critical for B cell development, specifically PAX5 and EBF1, cooperate with STAT5 activation to initiate ALL.


Fusaproliferin, a Fungal Mycotoxin, Shows Cytotoxicity against Pancreatic Cancer Cell Lines.
As a part of our ongoing research on endophytic fungi, we have isolated a sesterterpene mycotoxin, fusaproliferin (FUS), from a Fusarium solani strain, which is associated with the plant Aglaonema hookerianum Schott. FUS showed rapid and sub-micromolar IC50 against pancreatic cancer cell lines. Time-dependent survival analysis and microscopy imaging showed rapid morphological changes in cancer cell lines 4 h after incubation with FUS. This provides a new chemical scaffold that can be further developed to obtain more potent synthetic agents against pancreatic cancer.


Prevention of Noise-Induced Hearing Loss from Recreational Firearms.
In the United States and other parts of the world, recreational firearm shooting is a popular sport that puts the hearing of the shooter at risk. Peak sound pressure levels (SPLs) from firearms range from ∼140 to 175 dB. The majority of recreational firearms (excluding small-caliber 0.17 and 0.22 rifles and air rifles) generate between 150 and 165 dB peak SPLs. High-intensity impulse sounds will permanently damage delicate cochlear structures, and thus individuals who shoot firearms are at a higher risk of bilateral, high-frequency, noise-induced hearing loss (NIHL) than peer groups who do not shoot. In this article, we describe several factors that influence the risk of NIHL including the use of a muzzle brake, the number of shots fired, the distance between shooters, the shooting environment, the choice of ammunition, the use of a suppressor, and hearing protection fit and use. Prevention strategies that address these factors and recommendations for specialized hearing protectors designed for shooting sports are offered. Partnerships are needed between the hearing health community, shooting sport groups, and wildlife conservation organizations to develop and disseminate accurate information and promote organizational resources that support hearing loss prevention efforts.


The personal and social consequences of Alzheimer disease.
Alzheimer disease (AD) has a drastic effect on the lives of those affected by the disease. Surprisingly little research has studied the personal and social consequences for AD patients. Many appear to go through six stages of change as the disease progresses, but any individual's path through the disease may vary substantially. AD also has a drastic effect on family members who serve as caregivers for people with the disease. These effects are much more studied and better documented. The burdens and, in some cases, benefits of the caregiver role differ from person to person. Race and ethnicity are among the variables that seem to play a role in the caregiver's response. The economic costs of AD are considerable, both for "formal" services, paid for in the money economy, and "informal" services, provided without monetary payment. The direct costs, both formal and informal, are largely, but not entirely, borne by the patient and the patient's family. The overall costs to society clearly amount to scores of billions of dollars.


[Effect of lysosomal cathepsin D and acid phosphatase on collagen protein catabolism in cirrhotically altered liver after stimulating regeneration with choriogonin].
Dependence of collagen fractions content from the activities of cathepsin D and acid phosphatase as well as dynamics of their alteration under the influence of choriogonine were studied in cirrhotic liver tissue of rats. Content of the collagen proteins was decreased, with simultaneous increase in amount of their soluble fractions, after treatment of the animals by choriogonine. At the first step of the experiment the activity of the hydrolytic enzymes was decreased but it was normalized to the end of the experiment; at the same time, activity of cathepsin D remained high in the non-sedimentable fraction. Correlation between the activity of cathepsin D and content of collagen fractions was observed in the liver tissue. This enzyme appears to participate in catabolism of collagen.


The pharmacokinetics of proguanil in human subjects following a single oral dose.
The pharmacokinetics of orally administered 200-mg dose of proguanil in volunteers, one African and one Caucasian, is described. The drug was rapidly absorbed reaching a peak concentration in the blood within 3 h, and declining slowly thereafter to give a terminal phase elimination half life of 11.20 +/- 4.10 h and a systemic clearance of 1.270 +/- 0.020 l/h/kg. The small apparent volume of distribution shows that the drug is confined mainly to the blood and is not extensively bound to tissues; it undergoes cyclic oxidation in the liver to cycloguanil--the active metabolite responsible for antimalarial activity. Cycloguanil was detected in the plasma 3 h after proguanil ingestion and reached peak concentration between 5 h and 6 h. Excretion of proguanil was rapid, 60% of the single dose passing through the renal system within 24 h.


Tempo and mode of evolutionary radiation in iguanian lizards.
Identification of general properties of evolutionary radiations has been hindered by the lack of a general statistical and phylogenetic approach applicable across diverse taxa. We present a comparative analytical framework for examining phylogenetic patterns of diversification and morphological disparity with data from four iguanian-lizard taxa that exhibit substantially different patterns of evolution. Taxa whose diversification occurred disproportionately early in their evolutionary history partition more of their morphological disparity among, rather than within, subclades. This inverse relationship between timing of diversification and morphological disparity within subclades may be a general feature that transcends the historically contingent properties of different evolutionary radiations.


Global identifiability of nonlinear models of biological systems.
A prerequisite for well-posedness of parameter estimation of biological and physiological systems is a priori global identifiability, a property which concerns uniqueness of the solution for the unknown model parameters. Assessing a priori global identifiability is particularly difficult for nonlinear dynamic models. Various approaches have been proposed in the literature but no solution exists in the general case. In this paper, we present a new algorithm for testing global identifiability of nonlinear dynamic models, based on differential algebra. The characteristic set associated to the dynamic equations is calculated in an efficient way and computer algebra techniques are used to solve the resulting set of nonlinear algebraic equations. The algorithm is capable of handling many features arising in biological system models, including zero initial conditions and time-varying parameters. Examples of usage of the algorithm for analyzing a priori global identifiability of nonlinear models of biological and physiological systems are presented.


Dendrites impact the encoding capabilities of the axon.
This study highlights a new and powerful direct impact of the dendritic tree (the input region of neurons) on the encoding capability of the axon (the output region). We show that the size of the dendritic arbors (its impedance load) strongly modulates the shape of the action potential (AP) onset at the axon initial segment; it is accelerated in neurons with larger dendritic surface area. AP onset rapidness is key in determining the capability of the axonal spikes to encode (phase lock to) rapid changes in synaptic inputs. Hence, our findings imply that neurons with larger dendritic arbors have improved encoding capabilities. This "dendritic size effect" was explored both analytically as well as numerically, in simplified and detailed models of 3D reconstructed layer 2/3 cortical pyramidal cells of rats and humans. The cutoff frequency of spikes phase locking to modulated inputs increased from 100 to 200 Hz in pyramidal cells of young rats to 400-600 Hz in human cells. In the latter case, phase locking reached close to 1 KHz in in vivo-like conditions. This work highlights new and functionally profound cross talk between the dendritic tree and the axon initial segment, providing new understanding of neurons as sophisticated nonlinear input/output devices.


Y3IP1, a nucleus-encoded thylakoid protein, cooperates with the plastid-encoded Ycf3 protein in photosystem I assembly of tobacco and Arabidopsis.
The intricate assembly of photosystem I (PSI), a large multiprotein complex in the thylakoid membrane, depends on auxiliary protein factors. One of the essential assembly factors for PSI is encoded by ycf3 (hypothetical chloroplast reading frame number 3) in the chloroplast genome of algae and higher plants. To identify novel factors involved in PSI assembly, we constructed an epitope-tagged version of ycf3 from tobacco (Nicotiana tabacum) and introduced it into the tobacco chloroplast genome by genetic transformation. Immunoaffinity purification of Ycf3 complexes from the transplastomic plants identified a novel nucleus-encoded thylakoid protein, Y3IP1 (for Ycf3-interacting protein 1), that specifically interacts with the Ycf3 protein. Subsequent reverse genetics analysis of Y3IP1 function in tobacco and Arabidopsis thaliana revealed that knockdown of Y3IP1 leads to a specific deficiency in PSI but does not result in loss of Ycf3. Our data indicate that Y3IP1 represents a novel factor for PSI biogenesis that cooperates with the plastid genome-encoded Ycf3 in the assembly of stable PSI units in the thylakoid membrane.


Premarital sex behaviors among college youths of Kathmandu, Nepal.
Unhealthy premarital sex behavior leads to several health problems namely; sexually transmitted diseases (STDs), Human Immune Virus and Acquired Immune Deficiency Syndrome (HIV/AIDS), unwanted pregnancies, abortions and maternal deaths. Unwanted pregnancies and unsafe abortions are rampant in Nepal despite introduction of legal provision for safe abortion since 2003. Lately, unsafe sex and sex without condoms and drinking before sex is increasing in trend in youth population. The primary aim of this study was to explore the factors associated with premarital sex behaviors. The main objective of the study was to identify the premarital sex behaviors and related factors among college youths in Kathmandu, Nepal. Descriptive and explorative study of size 230 college youths aged between 18- 24 years. Data were collected by self-administered questionnaire from February 26 to March 15, 2012. The reliability of the questionnaire was ensured by using Cronbach's alpha. Late youths of age 20-24 were more likely to experience premarital sex than early youths of age 18-19 years old. Urban youths were less involved in premarital sex behavior than rural youth. Youths having negative attitudes towards premarital sex were more likely to experience premarital sex than a positive attitude. Youths who had good relationship with their parents had less premarital sex experience than youths having poor relationship with their parents. One fifth of college youth had premarital sex experience where alcohol drinker had higher premarital sex experience than non-drinker. Youths having good peer norms were significantly less likely to experience premarital sex behaviors than youths having poor peer norms.


Impact of Surgical Resection on Survival Outcomes After Chemoradiotherapy in Anal Adenocarcinoma.
Anal adenocarcinoma is a rare malignancy with a poor prognosis, and no randomized data are available to guide management. Prior retrospective analyses offer differing conclusions on the benefit of surgical resection after chemoradiotherapy (CRT) in these patients. We used the National Cancer Database (NCDB) to analyze survival outcomes in patients undergoing CRT with and without subsequent surgical resection. Patients with adenocarcinoma of the anus diagnosed in 2004 through 2015 were identified using the NCDB. Patients with metastatic disease and survival <90 days were excluded. We analyzed patients receiving CRT and stratified by receipt of surgical resection. Logistic regression was used to evaluate predictors of use of surgery and to form a propensity score-matched cohort. Overall survival (OS) was compared between treatment strategies using Cox proportional hazards regression. We identified 1,747 patients with anal adenocarcinoma receiving CRT, of whom 1,005 (58%) received surgery. Predictors of increased receipt of surgery included age <65 years, private insurance, overlapping involvement of the anus and rectum, N0 disease, and external-beam radiation dose ≥4,000 cGy. With a median follow-up of 3.5 years, 5-year OS was 61.1% in patients receiving CRT plus surgery compared with 39.8% in patients receiving CRT alone (log-rank P<.001). In multivariate analysis, surgery was associated with significantly improved OS (hazard ratio, -0.59; 95% CI, 0.50-0.68; P<.001). This survival benefit persisted in a propensity score-matched cohort (log-rank P<.001). In the largest series of anal adenocarcinoma cases to date, treatment with CRT followed by surgery was associated with a significant survival benefit compared with CRT alone in propensity score-matching analysis. Our findings support national guideline recommendations of neoadjuvant CRT followed by resection for patients with anal adenocarcinoma.


Type I interferon induced by TLR2-TLR4-MyD88-TRIF-IRF3 controls Mycobacterium abscessus subsp. abscessus persistence in murine macrophages via nitric oxide.
Mycobacterium abscessus (MAB) is an emerging, rapidly growing non-tuberculous Mycobacterium causing therapy-resistant pulmonary disease especially in patients with cystic fibrosis (CF). Smooth and rough colony type MAB can be isolated from infected patients whereby rough colony type MAB are more often associated with severe disease. Disease severity is also associated with an alternated type I interferon (IFN-I) response of the MAB-infected patients. However the relevance of this response for the outcome of MAB infection is still unknown. In this study, we analyzed the IFNβ expression of murine macrophages infected with a MAB rough colony strain (MAB-R) isolated from a patient with progressive CF and compared it to macrophages infected with the MAB smooth colony type reference strain (MAB-S). We found that MAB-R infected macrophages expressed significantly more IFNβ mRNA and protein than MAB-S infected macrophages. Higher IFNβ induction by MAB-R was associated with higher TNF expression and intracellular killing while low IFNβ induction was associated with lower TNF expression and persistence of MAB-S. IFNβ induction was independent of the intracellular cGAS-STING recognition pathway. MAB appeared to be recognized extracellularly and induced IFNβ expression via TLR2-TLR4-MyD88-TRIF-IRF3 dependent pathways. By using macrophages lacking the IFN-I receptor we demonstrate that MAB induced IFN-I response essentially contributed to restricting MAB-R and MAB-S infections by activating macrophage Nos2 expression and nitric oxide production. Thus IFN-I seem to influence the intrinsic ability of macrophages to control MAB infections. As MAB persists over long time periods in susceptible patients, our findings suggest that virulence of MAB strains is promoted by an insufficient IFN-I response of the host.


Transforming growth factor-beta1-induced Smad signaling, cell-cycle arrest and apoptosis in hepatoma cells.
Transforming growth factor-beta1 (TGFbeta) is involved in the regulation of liver cell proliferation and apoptosis, and escape of hepatoma cells from the growth restraining signals of TGFbeta has been suggested to contribute to tumor development. TGFbeta modulates gene transcription by receptor-mediated activation of Smad proteins which act as transcription factors. TGFbeta-mediated primary signaling responses as well as effects on the cell cycle and apoptosis were investigated in the human hepatoblastoma line HepG2, the rat hepatoma line FTO-2B and the mouse hepatoma line 55.1c. Activation of a Smad (Sma and Mad homolog) response-element-driven luciferase reporter by TGFbeta was very similar in all three cell lines, indicating functionality of the primary TGFbeta signaling pathway. Moreover, TGFbeta-inducible early gene was transiently activated by TGFbeta in all cell lines as shown by RT-PCR. HepG2 cells, however, were completely resistant to TGFbeta-induced growth arrest and apoptosis and 55.1c cells were only slightly susceptible to TGFbeta-induced apoptosis. By contrast, treatment of FTO-2B cells with TGFbeta led to a partial G0/G1 arrest and a strong induction of apoptosis. TGFbeta-induced apoptosis of FTO-2B cells was inhibited by dexamethasone, insulin, phenobarbital and dieldrin. Of these agents, only insulin led to a significant reduction of TGFbeta-stimulated Smad-reporter activity, suggesting that the other compounds interfere with TGFbeta-induced apoptosis downstream of Smad-mediated primary transcriptional responses at a level that may be constitutively altered in apoptosis-resistant hepatoma cell lines.


Specificity and 6-month durability of immune responses induced by DNA and recombinant modified vaccinia Ankara vaccines expressing HIV-1 virus-like particles.
Clade B DNA and recombinant modified vaccinia Ankara (MVA) vaccines producing virus-like particles displaying trimeric membrane-bound envelope glycoprotein (Env) were tested in a phase 2a trial in human immunodeficiency virus (HIV)-uninfected adults for safety, immunogenicity, and 6-month durability of immune responses. A total of 299 individuals received 2 doses of JS7 DNA vaccine and 2 doses of MVA/HIV62B at 0, 2, 4, and 6 months, respectively (the DDMM regimen); 3 doses of MVA/HIV62B at 0, 2, and 6 months (the MMM regimen); or placebo injections. At peak response, 93.2% of the DDMM group and 98.4% of the MMM group had binding antibodies for Env. These binding antibodies were more frequent and of higher magnitude for the transmembrane subunit (gp41) than the receptor-binding subunit (gp120) of Env. For both regimens, response rates were higher for CD4(+) T cells (66.4% in the DDMM group and 43.1% in the MMM group) than for CD8(+) T cells (21.8% in the DDMM group and 14.9% in the MMM group). Responding CD4(+) and CD8(+) T cells were biased toward Gag, and >70% produced 2 or 3 of the 4 cytokines evaluated (ie, interferon γ, interleukin 2, tumor necrosis factor α, and granzyme B). Six months after vaccination, the magnitudes of antibodies and T-cell responses had decreased by <3-fold. DDMM and MMM vaccinations with virus-like particle-expressing immunogens elicited durable antibody and T-cell responses.


Immunohistochemical evaluation of CD31 in human cystic radicular lesions and in keratocysts.
Platelet-endothelial cell adhesion molecule-1 protein (PECAM-1/CD31) is expressed in numerous physiological and pathological processes characterized by an increase of vascular permeability, and in normal and tumour tissues. CD31, member of the immunoglobulin super-family that mediates cell-to-cell adhesion, is a transmembrane glycoprotein, 130-140 kDa, also know as platelet-endothelium cell adhesion molecule (PECAM-1). CD31 is a ligand for CD38 and plays a role in thrombosis and angiogenesis. CD31 is strongly expressed in endothelial cells and weakly expressed in megakaryocytes, platelets, occasional plasma cells, lymphocytes (marginal zone B-cells, peripheral T-cells) and neutrophils. The present study evaluates the angiogenetic processes which are accompanied by an expansion of cystic radicular and keratocystic lesions of the jaw bone. Twelve subjects with maxillary cysts (8 males and 4 females) with an average age of 43 years were selected by the Chieti University Oral Surgery Department. The surgical samples taken were subjected to histological and immunohistochemical analysis. The histological evaluation confirmed the diagnosis of radicular cystisis and keratocystisis. The immunohistochemical analyses were positive for CD31 protein in all the lesions analysed, even though they had different intensities. Using a semiquantive analysis it was possible to highlight, in the radicular cyst samples, an intense expression of the vascular component both in the inflamed area and the adjacent stroma. The lesions with cheratin content showed newly-formed, rather modest, vascularity both in the area showing slight inflammation, where the cellular component is prevalent, and in the adjacent areas showing no sign of inflammation. Therefore, in our observations, angiogenesis could take on a primary role in the development of cystic lesions of the jaw bones. The differences of CD31 expression, in all samples, would advise for a wider monitoring able to evaluate the possible use of such a protein as a diagnostic marker.


Receptor for bombesin with associated tyrosine kinase activity.
The neuropeptide bombesin is known for its potent mitogenic activity on murine 3T3 fibroblasts and other cells. Recently it has been implicated in the pathogenesis of small cell lung carcinoma, in which it acts through an autocrine loop of growth stimulation. Phosphotyrosine (P-Tyr) antibodies have been successfully used to recognize the autophosphorylated receptors for known growth factors. In Swiss 3T3 fibroblasts, phosphotyrosine antibodies identified a 115,000-Mr cell surface protein (p115) that became phosphorylated on tyrosine as a specific response to bombesin stimulation of quiescent cells. The extent of phosphorylation was dose dependent and correlated with the mitogenic effect induced by bombesin, measured by [3H]thymidine incorporation. Tyrosine phosphorylation of p115 was detectable minutes after the addition of bombesin, and its time course paralleled that described for the binding of bombesin to its receptor. Immunocomplexes of phosphorylated p115 and phosphotyrosine antibodies bound 125I-labeled [Tyr4]bombesin in a specific and saturable manner and displayed an associated tyrosine kinase activity enhanced by bombesin. Furthermore, the 125I-labeled bombesin analog gastrin-releasing peptide, bound to intact live cells, was coprecipitated with p115. These data strongly suggest that p115 participates in the structure and function of the surface receptor for bombesin, a new member of the family of growth factor receptors with associated tyrosine kinase activity.


Intensive care unit antimicrobial resistance and the role of the pharmacist.
Over the past 20 yrs, pharmacists have successfully integrated their services and expertise to gain acceptance as full members of pediatric, surgical, medical, and intensive care unit (ICU) patient care teams. The pharmacists' training in pharmacology, pharmacokinetics, pharmacodynamics, and pharmacoeconomics complements the expertise of other members of the patient care team. Generally, a strong background in infectious diseases and critical care also provides a focal point for clinical pharmacy service intervention. Although practitioners often focus on issues exclusively related to their specific hospital or ICU, the issues surrounding antibiotic resistance are more global and societal in nature. Medical, surgical, and pharmaceutical practices inside the hospital and ICU extend their influence into the community. Customs and practices of daily living in our society coupled with use of agents capable of altering microbial flora impact our hospital and ICU when patients from the community are admitted. The misuse of antibiotics and the lack of effective infection control programs are often identified as key components in the perpetuation of these phenomena. The focus for the pharmacist and the ICU team must be on the optimization of antibiotic use and infection control guidelines. This review will address the many issues that surround the appropriate use of antibiotics and what role the pharmacist can play in ensuring the optimal use of infection control measures in the ICU and hospital.


Fresh frozen plasma transfusion after cardiac surgery.
Fresh frozen plasma (FFP) transfusion in the intensive care unit (ICU) is commonly used to treat coagulopathy and bleeding in cardiac surgery, despite suggestion that it may increase the risk of morbidity and mortality through mechanisms such as fluid overload and infection. We retrospectively studied consecutive adults undergoing cardiac surgery from the Medical Information Mart for Intensive Care III and IV databases. We applied propensity score matching to investigate the independent association of within-ICU FFP transfusion with mortality and other key clinical outcomes. Of our 12,043 adults who met inclusion criteria, 1585 (13.2%) received perioperative FFP with a median of 2.48 units per recipient (interquartile range [IQR]: 2.04, 4.33) at a median time of 1.83 h (IQR: 0.75, 3.75) after ICU admission. After propensity matching of 952 FFP recipients to 952 controls, we found no significant association between FFP use and hospital mortality (odds ratio (OR): 1.58; 99% confidence interval (CI): 0.57, 3.71), suspected infection (OR: 0.72; 99% CI: 0.49, 1.08), or acute kidney injury (OR: 1.23; 99% CI: 0.91, 1.67). However, FFP was associated with increased days in hospital (adjusted mean difference (AMD): 1.28; 99% CI: 0.27, 2.41; p = .0050), days in intensive care (AMD: 1.28; 99% CI: 0.27, 2.28; p = .0011), and chest tube output in millilitres up to 8 h after transfusion (AMD: 92.98; 99% CI: 52.22, 133.74; p < .0001). After propensity matching, FFP transfusion was not associated with increased hospital mortality, but was associated with increased length of stay and no decrease in bleeding in the early post-transfusion period.


Derivation of smooth muscle cells with neural crest origin from human induced pluripotent stem cells.
The heterogeneity of vascular smooth muscle cells (SMCs) is related to their different developmental origins such as the neural crest and mesoderm. Derivation of SMCs from different origins will provide valuable in vitro models for the investigation of vascular development and diseases. From the perspective of regenerative medicine and tissue engineering, an expandable cell source of SMCs is required for the construction of tissue-engineered blood vessels. In this study, we developed a robust protocol to derive neural crest stem cells (NCSCs) from human embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs). NCSCs derived from ESCs and iPSCs were expandable with similar cell doubling times. NCSCs were capable of differentiating into neural and mesenchymal lineages. TGF-β1 induced the expression of SMC markers calponin-1, SM22α, and smooth muscle myosin heavy chain and resulted in the assembly of smooth muscle α-actin, calponin-1, and SM22α into stress fibers. This work provides a basis for using iPSCs to study SMC biology and deriving vascular cells for tissue engineering.


Changing spectrum of primary pyomyositis in North India.
Recent data have demonstrated the changing epidemiology of primary pyomyositis worldwide. Our hospital-based retrospective study investigated the clinical and microbiological spectrum of primary pyomyositis between 2013 and 2021 in PGIMER (Chandigarh), India. Over a quarter had predisposing conditions, mainly diabetes mellitus and immunosuppressive therapy. Fever, muscle pain, local swelling and breathlessness were the usual presentations, with quadriceps, iliopsoas and gluteal muscles commonly affected. Staphylococcus aureus was the predominant cause, with c.50% methicillin-resistant strains. Almost two-thirds presented with metastatic infection (stage 3 pyomyositis), frequently with septic lung emboli. Patients with methicillin-sensitive and resistant Staphylococcus aureus had a similar incidence of metastatic infection. In-hospital mortality was c.10% and was strongly associated with a high international normalised ratio. Primary pyomyositis remains a significant problem, with a dramatic increase in community-associated methicillin-resistant Staphylococcus aureus.


Acute health hazards of firefighters after fighting a department store fire.
The purpose of this study was to evaluate the health hazards of firefighters after fighting a department store fire which lasted for 40 hours. Respiratory symptoms of 168 firefighters were collected and the pulmonary functions of 149 firefighters were measured by spirometer two days after fighting the fire and compared to 32 controls. The principal symptoms manifested by firefighters were burning eyes and mucous membrane irritation. Cross-sectional study of the pulmonary functions showed that FEV1.0, FEV1.0/FVC and the flow rates (MMF, FEF25%, and FEF75%) in smoking exposed firefighters were significantly lower than those in smoking controls. However, only FEV1.0 (%) was significantly lower in nonsmoking exposed firefighters than in nonsmoking controls. Both FVC and FEV1.0 showed a declining trend with the duration of fire fighting. This study provides further evidence that firefighters are exposed to irritants during fighting a fire. Exposure to the combustion products could lead to pulmonary function defects, especially in smoking firefighters.


Tobacco use among teachers [corrected] in Uttar Pradesh & Uttaranchal, India.
Information about tobacco use prevalence, knowledge and attitude was assessed among school personnel in Uttar Pradesh. A single cluster sample design with probability proportional to the enrolment in grades 8-10 was used. Statistical analysis was done using SUDAAN and the C-sample procedure in Epi-Info. The school response rate was 100%. School personnel response rate ranged from 72-80%, the proportion of men being 84-92%. Current cigarette smoking and smokeless tobacco use reported by all teachers was 21.9% and 75.6% respectively. The prevalence of daily cigarette smoking was ranged by 12.6-15.1%; bidi and other smoking 4.8-13.4%; smokeless tobacco use 16.3-19.8%. Existing school policy on four measures were reported poor however over 72% school personnel felt need for policy prohibiting tobacco use among students and school personnel. Tobacco prevention instruction by teachers did not fare much better on six different measures (4.9-30.9%). However over 2/3rd school personnel were very supportive on tobacco control issues. There was no training among school personnel on tobacco use prevention skills (3.7%). However most of the school personnel (67.1%) were curious about getting such trainings. A positive environment for tobacco use prevention needs to be created by adopting comprehensive tobacco control policies for schools. First step towards this may be training of school personnel on tobacco use prevention skill and supply of teaching materials.


Acute effects of mifepristone on emotional processing related brain activity: A functional MRI study.
The Hypothalamic-pituitary-adrenal (HPA) axis plays an important role in the pathophysiology of mood disorders, and preliminary data suggests that glucocorticoid receptor (GR) antagonism may be an important therapeutic mechanism. The effects of modulating HPA axis function on emotional processing related brain activity, which may be abnormal in depressed mood, is poorly understood. This study used a pharmacological functional magnetic resonance imaging (fMRI) design to determine the effects of the GR and progesterone receptor antagonist mifepristone on emotional faces processing task related brain activations in 19 right-handed healthy male participants. Each participant received 600 mg mifepristone or placebo on two separate imaging days and then performed an emotional processing fMRI task four hours later. The effect of mifepristone on task related brain activations was determined using Region-of-Interest (ROI) analyses and an exploratory whole brain voxel-wise analyses. No significant changes were observed in the defined ROIs (amygdala, anterior cingulate cortex, insula) or in the exploratory whole brain analyses that was associated with mifepristone administration in either the angry vs happy faces or angry and happy faces vs implicit baseline contrasts. Task reaction times and accuracy were similar in both mifepristone and placebo conditions (all p > 0.05). Our study failed to show significant evidence of modulation of emotional processing related brain activity associated with acute mifepristone administration. Future research should use fMRI to investigate the longer-term administration effects of mifepristone on mood in healthy participants and people with mood disorders to provide a deeper understanding of the potential effects on depressive symptoms.


Genetic diagnosis of band 3 deficiency using a quenching probe (QProbe)-PCR assay in bovine embryos.
The present study was conducted to develop a simple and rapid procedure to determine the genotype of band 3 deficiency in bovine embryos by a novel real-time PCR method using a fluorescent quenching-based probe (QProbe-PCR). QProbe-PCR successfully distinguished wild type and R664X mutant alleles by melting curve analysis. Minimal amounts of DNA template were required for the detection of wild type/wild type alleles, mutant/mutant alleles, and wild type/mutant alleles; their amounts were 10 pg, 25 pg, and 50 pg, respectively. When 10 blastomeres were used as a DNA sample, accuracies of genotyping by QProbe-PCR were 100% and 89% in embryos homozygous for the wild type allele and heterozygous for the wild type and mutant alleles, respectively. QProbe-PCR takes approximately 2 h for genotyping and requires lesser time than the conventional method using PCR-RFLP, which requires digestion with a restriction enzyme and electrophoresis. Our data showed that QProbe-PCR is a useful method for rapid analysis of the genetic deficiency in preimplantation embryos. Reduction in the time required for genotyping enabled the transfer of genetically selected embryos to recipient cows on the day of embryo collection. These results suggest that determination of the genotype for the genetic deficiency in embryos is useful to select animals free from the genetic disease, and it also makes it possible to produce an animal model homozygous for the mutation.


Mammalian recombination hot spots: properties, control and evolution.
Recombination, together with mutation, generates the raw material of evolution, is essential for reproduction and lies at the heart of all genetic analysis. Recent advances in our ability to construct genome-scale, high-resolution recombination maps and new molecular techniques for analysing recombination products have substantially furthered our understanding of this important biological phenomenon in humans and mice: from describing the properties of recombination hot spots in male and female meiosis to the recombination landscape along chromosomes. This progress has been accompanied by the identification of trans-acting systems that regulate the location and relative activity of individual hot spots.


PPARγ and chronic kidney disease.
Peroxisome proliferator-activated receptor-γ (PPARγ) agonists, exemplified by the thiazolidinediones (TZDs), have been used extensively for their beneficial effects to improve insulin sensitivity and lipid metabolism in type 2 diabetic patients. PPARγ receptors are part of the steroid hormone nuclear receptor family and, when activated by agonist binding, can affect numerous target genes expressing PPAR response elements. Results from experimental studies and a limited number of studies in humans suggest that PPARγ agonists have manifold effects beyond those on dysmetabolic syndrome. These potentially beneficial actions are mediated via renal parenchymal and infiltrating cells and modulate fibrotic, inflammatory, immune, proliferative, reactive oxygen and mitochondrial injury pathways. Thus, the potential benefits of TZDs in chronic kidney disease impact numerous pathogenic pathways. This review will focus on evidence of the effects of TZDs in nondiabetic chronic kidney disease in experimental and human disease settings.


[The influence of initial values of donor platelets count on the efficiency of plateletpheresis with the cell separator Fenval Baxter Amicus s.v.2.51].
The collection of platelets by apheresis is considered as a very great progress in transfusional medicine. A larger yield (total number of collected platelets) is obtained if the donor has a greater number of initial platelets and if the separation is done in a shorter time. One of the parameters is also the efficiency of the platelet collection (expressed in percentage) on the value of which different factors may have direct or indirect influence. To calculate the efficiency of platelet collection with the separator Fenval Baxter AMICUS s.v.2.51 and to compare the efficiency of platelet collection with this separator in relation to the initial value of donor platelets. The donors who participated in this study were divided into groups according to the value of the donors' number of platelets before separation. Group A consisted of donors whose initial value of the platelet number was higher than 200 but lower than 300 x 109/L Group B consisted of donors whose initial platelet number was higher than 300 but less than 450 x 109/L. The process was carried out on Fenval Baxter Amicus s.v.2.51. The expected efficiency of the collection was obtained by dividing the total number of collected cells by the expected total number of processed cells, i.e., the total number of cells passed through the equipment. In the 258 separations which satisfied the fixed criteria the donors were men in 226 cases (87.6%) and women in 32 (12.4%). The efficiency of platelet collection in our material gave 64.14 +/- 7.92 in the range of minimal and maximal value from 36.65 to 85.22. There is no relation in the correlation between the efficiency of the initial value of the number of platelets. There is no statistical significance in the difference in efficiency of the collection of platelets in the groups A and B. The separations lasted for 44.72 (+/- 9.35) minutes, in group A 49.40 (7.30) minutes and (+/- 8.27) in group B. There is a significant statistical difference (p 0.050; Equal Variance Test P = 0.527). The values we have obtained from our material are similar to results from other studies of this procedure with one important difference which is that our results were obtained from a larger number of samples. We did not find any relation in the correlation in the variables, the efficiency of platelet separation or in the value of initial number of platelets. The efficiency of platelet collection is almost the same in groups A and B (64.65 +/- 7.99 in group A and 63.50 +/- 7.83 in group B and there is no statistically significant difference). The difference in time of the separation is statistically significant in the groups A and B. The initial value of the number of the donor's platelets has no effect on the efficiency of platelet collection by apheresis but they have effect on the duration of the separation and the value of the yield. With a higher number of the value of platelets we can cut the time of the separation and obtain a greater yield. It is preferable to choose donors with a greater value of the initial number of platelets. In this way it is possible to obtain a greater yield, cut the time of separation and improve the efficiency of platelet collection. This is beneficial both from the medical point of view because of the greater yield and from the financial point of view because of the shorter time of separation and greater efficiency.


Batch-Mode Clinical-Scale Optical Hyperpolarization of Xenon-129 Using an Aluminum Jacket with Rapid Temperature Ramping.
We present spin-exchange optical pumping (SEOP) using a third-generation (GEN-3) automated batch-mode clinical-scale 129Xe hyperpolarizer utilizing continuous high-power (∼170 W) pump laser irradiation and a novel aluminum jacket design for rapid temperature ramping of xenon-rich gas mixtures (up to 2 atm partial pressure). The aluminum jacket design is capable of heating SEOP cells from ambient temperature (typically 25 °C) to 70 °C (temperature of the SEOP process) in 4 min, and perform cooling of the cell to the temperature at which the hyperpolarized gas mixture can be released from the hyperpolarizer (with negligible amounts of Rb metal leaving the cell) in approximately 4 min, substantially faster (by a factor of 6) than previous hyperpolarizer designs relying on air heat exchange. These reductions in temperature cycling time will likely be highly advantageous for the overall increase of production rates of batch-mode (i.e., stopped-flow) 129Xe hyperpolarizers, which is particularly beneficial for clinical applications. The additional advantage of the presented design is significantly improved thermal management of the SEOP cell. Accompanying the heating jacket design and performance, we also evaluate the repeatability of SEOP experiments conducted using this new architecture, and present typically achievable hyperpolarization levels exceeding 40% at exponential build-up rates on the order of 0.1 min-1.


Facial nerve repair accomplished by the interposition of a collagen nerve guide.
Facial nerve paralysis due to a surgical procedure or trauma is a frequently observed complication. The authors evaluated facial nerve repair achieved by the interposition of a collagen nerve guide. Ten cats were divided into three groups. Group 1 consisted of six animals in which a 5-mm facial nerve segment on one side was resected and replaced by a collagen tube that was sutured to bridge both nerve stumps. On the opposite side a 5-mm segment of facial nerve was resected, reversed 180 degrees, and sutured to the stumps as an autograft nerve. Group 2 consisted of two cats in which the collagen nerve guide was interposed on one side and the nerve on the other side was left intact. Group 3 consisted of two cats in which a reversed autograft nerve was placed on one side and the nerve on the other side was left intact. Histological, electrophysiological, and horseradish peroxidase labeling examinations were performed starting 3 weeks after surgery. Light and electron microscopic examinations of collagen tube-implanted specimens revealed a well-vascularized regenerated nerve. The electrophysiological study confirmed the recovery of electrical activity in regenerated axons. Horseradish peroxidase labeling also confirmed restoration of the whole facial nerve tract. The collagen nerve guide shows great promise as a nerve conduit.