Protective Effects of Ferulic Acid against Chronic Cerebral Hypoperfusion-Induced Swallowing Dysfunction in Rats.
Ferulic acid (FA), a phenolic phytochemical, has been reported to exert antioxidative and neuroprotective effects. In this study, we investigated the protective effects of FA against the dysfunction of the swallowing reflex induced by ligation of bilateral common carotid arteries (2VO) in rats. In 2VO rats, topical administration of water or citric acid to the pharyngolaryngeal region evoked a diminished number of swallowing events with prolonged latency compared to sham-operated control rats. 2VO rats had an increased level of superoxide anion radical, and decreased dopamine and tyrosine hydroxylase enzyme levels in the striatum, suggesting that 2VO augmented cerebral oxidative stress and impaired the striatal dopaminergic system. Furthermore, substance P (SP) expression in the laryngopharyngeal mucosa, which is believed to be positively regulated by dopaminergic signaling in the basal ganglia, was decreased in 2VO rats. Oral treatment with FA (30 mg/kg) for 3 weeks (from one week before 2VO to two weeks after) improved the swallowing reflex and maintained levels of striatal dopamine and laryngopharyngeal SP expression in 2VO rats. These results suggest that FA maintains the swallowing reflex by protecting the dopamine-SP system against ischemia-induced oxidative damage in 2VO rats.


First steps towards effective methods in exploiting high-throughput technologies for the determination of human protein structures of high biomedical value.
The EC 'Structural Proteomics In Europe' contract is aimed specifically at the atomic resolution structure determination of human protein targets closely linked to health, with a focus on cancer (kinesins, kinases, proteins from the ubiquitin pathway), neurological development and neurodegenerative diseases and immune recognition. Despite the challenging nature of the analysis of such targets, approximately 170 structures have been determined to date. Here, the impact of high-throughput technologies, such as parallel expression of multiple constructs, the use of standardized refolding protocols and optimized crystallization screens or the use of mass spectrometry to assist sample preparation, on the structural biology of mammalian protein targets is illustrated through selected examples.


Development of a placebo effect model combined with a dropout model for bipolar disorder.
The aim of this study was to develop a placebo model for bipolar disorder to help optimize clinical trial designs for studies targeting manic episodes in bipolar disorder. A bipolar disease database was built based on individual longitudinal data collected from over 3,000 patients in 11 clinical trials for 5 approved bipolar drugs. An empirical placebo effect model with an exponential decay process plus a linear progression process was developed to quantify the time course of the Young Mania Rating Scale total score based on only placebo data from the database. In order to describe the dropout pattern during the trials, a parametric survival model was developed and the Weibull distribution was identified to be the best distribution to describe the data. Based on the likelihood ratio test, it was found that patients with higher baseline score, slower disease improvement and more rapid disease progression tended to dropout earlier, and the trial features such as trial starting year and trial site were also significant covariates for dropout. A combination of the placebo effect model and the dropout model was applied to simulate new clinical trials through Monte-Carlo simulation. Both the placebo effect model and dropout model described the observed data reasonably well based on various diagnostic plots. The joint placebo response and dropout models can serve as a tool to simulate the most likely level of placebo response with the expected dropout pattern to help design a new clinical trial.


A nasal osteoma with an acute course in a Japanese Black heifer.
A 14-month-old Japanese Black heifer presented with unilateral epistaxis and mild swelling of the right face. Radiography revealed a mass with increased radiopacity on the right side of the nasal bridge, extending to the left side. Intranasal endoscopy confirmed a large tumor-like structure protruding into the nasal cavity. Following euthanasia, cranial computed tomography (CT) was performed, revealing a tumor 24.3 × 17.5 × 14.8 cm in size. The tumor occupied the entire right nasal cavity and the frontal and sphenoid sinuses. Histopathological examination revealed that the tumor consisted of well-differentiated trabecular bones and loose connective tissue. Based on these findings, a diagnosis of osteoma was established. This report describes a case of osteoma with an acute course in a Japanese Black heifer.


Nanostructured peptide fibrils formed at the organic-aqueous interface and their use as templates to prepare inorganic nanostructures.
Formation of fibril-type nanostructures of the Alzheimer's beta-amyloid diphenylalanine (l-Phe-l-Phe, FF) at the organic-aqueous interface and the factors affecting their structures have been investigated. Such nanostructures are also formed by bovine serum albumin and bovine pancreas insulin. The concentration of the precursor taken in the aqueous layer plays an important role in determining the morphology of the nanostructures. The addition of curcumin to the organic layer changes the structure of the self-assembled one-dimensional aggregates of diphenylalanine. By coating the diphenylalanine dipeptide fibrils with appropriate precursors followed by calcination in air, it has been possible to obtain one-dimensional nanostructures of inorganic materials.


[Soil organic carbon dynamics and the prediction of their spatial changes in response to anthropogenically introduced shrub encroachment in desert steppe of the Eastern Ningxia, China.]
With the 14-year enclosed grassland and the grazed grassland as control, the impacts of anthropogenic shrublands (Caragana korshinskii) with the different planting years (3, 12, 22 a) and planting spaces (2, 8, 40 m) on soil organic carbon (SOC) contents were examined in the desert steppe of Eastern Ningxia, China. We further analyzed the spatial pattern and heterogeneity of SOC in 0-40 cm soil layer of the grassland area with introduced shrubs. The results showed that SOC in C. korshinskii shrublands had an increase trend with increased planting years and decreased spaces. The mean SOC with different planting years and spaces was 42.7% and 32.8% more than that in grazing land, respectively. There was no significant difference of SOC between shrublands and the 14-year enclosed grassland. The increase trend of SOC decreased by 27.0% in 22-year planting shrubland. The SOC content of 0-40 cm soil layer varied from 0.21 g·kg-1 to 26.04 g·kg-1 (with a mean of 3.75 g·kg-1), and the coefficient of SOC variation ranged from 90.9% to 114.7%. The SOC in 0-5 cm and 15-40 cm soil layers fitted the optimal theory formulation of Gaussian model, while that in 5-15 cm soil layer fitted a spherical model. The ranges (A0) of spatial autocorrelation in the 0-5 and 5-15 cm soil layers were smaller (3.11, 3.00 km) than that in 15-40 cm soil layer (10.10 km). The nugget/sill C0/(C0+C) of SOC in 0-5, 5-15 cm soil layer was 0.2% and 16.3%, indicating a strong spatial correlation, while that in 15-40 cm soil layer was 36.9%, with a moderate correlation. The shrub introdution could significantly accelerate the accumulation and fixation of SOC in top 40 cm soil layer in degraded desert steppe, but also intensified the spatial heterogeneity and SOC fragmentation. The SOC content in the anthropogenic shrublands had no significant difference from that in the enclosed desert steppe (14 years). The SOC spatial heterogeneity and the degree of fragmentation were weakened and decreased with the increasing soil layer depth.


Histone post-translational modifications to target memory-related diseases.
Epigenetic mechanisms, i.e. the control gene of expression without changing DNA sequence, include DNA methylation, histone post-translational modifications (PTMs) and microRNAs (miRNAs). Aberrant epigenetic modifications are associated with several pathological conditions, including brain diseases, resulting from environmental causes, ageing or genetic factors. The role of histone PTMs, including acetylation, phosphorylation, methylation and ubiquitylation, has been demonstrated in learning and memory, both in physiological conditions and in neuropathologies. Abnormalities in these modifications or in the machinery that control them are associated with several neurodegenerative, neuro-developmental and psychiatric diseases. Therefore, these epigenetic marks are promising targets to address memory-related diseases and strong efforts are presently focused on pharmacological and genetic approaches to this field.


NIHSS training and certification using a new digital video disk is reliable.
NIH Stroke Scale certification is required for participation in modern stroke clinical trials and as part of good clinical care in stroke centers. The existing training and certification videotapes, however, are more than 10 years old and do not contain an adequate balance of patient findings. After producing a new NIHSS training and demonstration DVD, we selected 18 patients representing all possible scores on 15 scale items for a new certification DVD. Patients were divided into 3 certification groups of 6 patients each, balanced for lesion side, distribution of scale item findings, and total score. We sought to measure interrater reliability of the certification DVD using methodology previously published for the original videotapes. Raters were recruited from 3 experienced stroke centers. Each rater watched the new training DVD and then evaluated one of the 3 certification groups. Responses were received from 112 raters: 26.2% of all responses came from stroke nurses, 34.1% from emergency departments/other physicians, and 39.6% from neurologists. One half (50%) of raters were previously NIHSS-certified. Item responses were tabulated, scoring performed as previously published, and agreement measured with unweighted kappa coefficients for individual items and an intraclass correlation coefficient for the overall score. kappa ranged from 0.21+/-0.05 (ataxia) to 0.92+/-0.09 (LOC-C questions). Of 15 items, 2 showed poor, 11 moderate, and 2 excellent agreement based on kappa scores. The intraclass correlation coefficient for total score was 0.94 (95% confidence interval, 0.84 to 1.00). Reliability scores were similar among specialists and centers, and there were no differences between nurses and physicians. kappa scores trended higher among raters previously certified. These certification DVDs are reliable for NIHSS certification, and scoring sheets have been posted on a web site for real-time, online certification.


Downsizing health coverage.
The crushing cost of insurance is fundamentally changing U.S. health care. Companies shift costs to employees, but low-wage workers often can't absorb them. Hospitals struggle to provide equitable care to all in the face of a payment system that seems to encourage rationing by income and insurance status. Have we reached the tipping point?


The Impact of Single Session Intermittent Theta-Burst Stimulation over the Dorsolateral Prefrontal Cortex and Posterior Superior Temporal Sulcus on Adults with Autism Spectrum Disorder.
Intermittent theta burst stimulation (iTBS), a patterned repetitive transcranial magnetic stimulation, was applied over the posterior superior temporal sulcus (pSTS) or dorsolateral prefrontal cortex (DLPFC) to explore its impact in adults with autism spectrum disorder (ASD). Among 25 adults with ASD, 19 (mean age: 20.8 years) completed the randomized, sham-controlled, crossover trial. Every participant received iTBS over the bilateral DLPFC, bilateral pSTS and inion (as a sham control stimulation) in a randomized order with a 1-week interval. Neuropsychological functions were assessed using the Conners' Continuous Performance Test (CCPT) and the Wisconsin Card Sorting Test (WCST). Behavioral outcomes were measured using the Yale-Brown Obsessive Compulsive Scale (Y-BOCS) and the Social Responsiveness Scale (SRS). In comparison to that in the sham stimulation, the reaction time in the CCPT significantly decreased following single DLPFC session (p = 0.04, effect size = 0.71) while there were no significant differences in the CCPT and WCST following single pSTS session. Besides, the results in behavioral outcomes were inconsistent and had discrepancy between reports of parents and patients. In conclusion, a single session of iTBS over the bilateral DLPFC may alter the neuropsychological function in adults with ASD. The impacts of multiple-sessions iTBS over the DLPFC or pSTS deserve further investigations.


Identification of Greb1l as a genetic determinant of crisscross heart in mice showing torsion of the heart tube by shortage of progenitor cells.
Despite their burden, most congenital defects remain poorly understood, due to lack of knowledge of embryological mechanisms. Here, we identify Greb1l mutants as a mouse model of crisscross heart. Based on 3D quantifications of shape changes, we demonstrate that torsion of the atrioventricular canal occurs together with supero-inferior ventricles at E10.5, after heart looping. Mutants phenocopy partial deficiency in retinoic acid signaling, which reflect overlapping pathways in cardiac precursors. Spatiotemporal gene mapping and cross-correlated transcriptomic analyses further reveal the role of Greb1l in maintaining a pool of dorsal pericardial wall precursor cells during heart tube elongation, likely by controlling ribosome biogenesis and cell differentiation. Consequently, we observe growth arrest and malposition of the outflow tract, which are predictive of abnormal tube remodeling in mutants. Our work on a rare cardiac malformation opens novel perspectives on the origin of a broader spectrum of congenital defects associated with GREB1L in humans.


[Molecular characterization of vancomycin-resistant Enterococci].
To investigate the homology and resistant mechanism of vancomycin-resistant Enterococci (VRE) isolates. A total of 9 VRE isolates were collected from 2006 to 2007 at PUMC hospital. The susceptibility of these isolates to 10 different antibiotics including vancomycin was tested by E-test. These strains were processed by brain heart infusion agar screening in the presence of vancomycin (6 microg/ml), and were analyzed for genotypic characteristics using the multiplex PCR. The homology of the isolates was determined by pulsed-field gel electrophoresis (PFGE). All the 9 VRE isolates were identified as Enterococci faecium. The visual analysis of PFGE patterns revealed 6 different PFGE types. The vanA gene was confirmed by PCR and sequencing in 9 VRE isolates, which were consistent between phenotype and genotype for glycopeptides resistance. Only vanA genotype was detected in PUMC hospital. Clonal dissemination, horizontal gene transfer, and the selective pressure of antimicrobial agents may contribute to the increase of VRE.


Mechanisms of intravitreal toxicity of indocyanine green dye: implications for chromovitrectomy.
Indocyanine green (ICG) dye was shown to improve the visualization of preretinal tissues during chromovitrectomy. However, controversy arose regarding the safety of intravitreal ICG application, because worse functional outcomes and a higher incidence of retinal pigment epithelium (RPE) changes and visual field defects were reported. The mechanisms of ICG-related toxicity and their relevance for chromovitrectomy are reviewed. A literature search was performed from 1998 through 2005 for relevant information related to the mechanisms of intravitreal ICG toxicity. Animal and clinical data on intravitreal ICG-related toxicity were collected to clarify the mechanisms of the risk of intravitreal ICG injection. Over 80 controversial in vitro, ex vivo, and in vivo animal investigations as well as clinical reports on intravitreal ICG staining were found in the literature. The main postulated mechanisms of intravitreal ICG-related toxicity were as follows: biochemical direct injury to the ganglion cells/neuroretinal cells, RPE cells, and superficial retinal vessels; apoptosis and gene expression alterations to either RPE cells or neuroretinal cells; osmolarity effect of ICG solution on the vitreoretinal interface; light-induced injury; and mechanical cleavage effect to the internal limiting membrane/inner retina. Whereas the exact mechanism of intravitreal ICG-related damage remains yet to be determined, most animal experiments proposed that ICG dye has a dose-dependent toxic effect on retinal tissue. This hypothesis was supported by clinical data, because better functional outcomes were obtained when low dye concentrations and short incubation times were reported. Much evidence supports that ICG dye has a dose-dependent toxic effect on the retina. Therefore, the following recommendations to minimize toxic effects on the retina are proposed: dye injection in concentrations as low as possible; avoidance of repeated ICG injections onto bare retina; dye injection far from the macular hole to prevent direct dye contact with the RPE; short incubation time of ICG in the vitreous cavity to diminish the concentration in contact with the retinal tissue; and the light pipe kept far from the retina throughout the whole surgical procedure.


COVID-19 Hyperinflammation: What about Neutrophils?
COVID-19 is often related to hyperinflammation that drives lung or multiorgan injury. The immunopathological mechanisms that cause excessive inflammation are under investigation and constantly updated. Here, a gene network approach was used on recently published data sets to identify possible COVID-19 inflammatory mechanisms and bioactive genes. First, network analysis of putative SARS-CoV-2 cellular receptors led to the mining of a neutrophil-response signature and relevant inflammatory genes. Second, analysis of RNA-seq data sets of lung cells infected with SARS-CoV-2 revealed that infected cells expressed neutrophil-attracting chemokines. Third, analysis of RNA-seq data sets of bronchoalveolar lavage fluid cells from COVID-19 patients identified upregulation of neutrophil genes and chemokines. Different inflammatory genes mined here, including TNFR, IL-8, CXCR1, CXCR2, ADAM10, GPR84, MME, ANPEP, and LAP3, might be druggable targets in efforts to limit SARS-CoV-2 inflammation in severe clinical cases. The possible role of neutrophils in COVID-19 inflammation needs to be studied further.


Carriage of methicillin-resistant Staphylococcus aureus at hospital admission.
To measure the prevalence of, and to establish predictors for, the nasal carriage of methicillin-resistant Staphylococcus aureus (MRSA) at hospital admission. To evaluate mannitol-salt agar with oxacillin for the simultaneous detection and identification of MRSA from nasal swabs. Three-month prospective case-control survey, with data collected from interviews and computerized databases. The criterion standard for MRSA detection was culture on Mueller-Hinton agar with oxacillin 6 microg/mL (National Committee for Clinical Laboratory Standards method). 320-bed tertiary-care hospital. 387 patients screened within 24 hours after admission, including 10 MRSA carriers (cases), 291 patients with no S aureus, and 86 patients with methicillin-susceptible S aureus. The prevalence of MRSA nasal carriage was 2.6%, whereas the prevalence of carriage was 3.1% when both nasal and wound cultures were performed. The significant predictors of carriage were a prior detection of MRSA, open wounds, diabetes mellitus, treatments by injection, prior nursing home stays, visits at home by a nurse, and prior antibiotic treatments. Cases had stayed for longer periods in hospitals and had received longer antibiotic treatments within a year. Eighty patients (including the 10 cases) had diabetes, had been exposed to healthcare facilities within a year, and had antibiotics within 6 months. The sensitivity and negative predictive value of nasal swabs on mannitol-salt agar with oxacillin were 60% and 71%, respectively. MRSA carriage on admission to the hospital may be an increasing and underestimated problem. Further studies are needed to develop and validate a sensitive and specific prediction rule.


Bioimpedance monitoring: better than chest x-ray for predicting abnormal pulmonary fluid?
Bioimpedance monitoring may aid in treating heart failure. Mean thoracic electrical impedance (Zo) is inversely proportional to thoracic fluid volume and may offer greater sensitivity for detecting thoracic fluid. OBJECTIVE. Compare bioimpedance monitoring thoracic fluid detection to that of chest x-ray. METHOD. Prospective convenience sample. SETTING. 1000 bed teaching hospital. PARTICIPANTS. Patients with suspected heart failure and shortness of breath. A single blinded radiologist interpreted chest x-rays as: normal, cardiomegaly, or abnormal pulmonary fluid. STATISTICS. General linear model with post hoc Bon Ferroni pairwise comparisons. RESULTS. 131 patients, mean age 66.8 years, 64.3% male, with an initial mean Zo=18 ohms. There was a significant difference (p<0.0002) between patients with cardiomegaly (Zo=17.5+/-5.5) or abnormal pulmonary fluid on chest x-ray (Zo=17.2+/-4.2) compared to normals (Zo=23.4+/-5.4). There was no difference between cardiomegaly and abnormal pulmonary fluid patients. CONCLUSION. Bioimpedance measurement may detect pulmonary fluid not apparent on chest radiograph. (c)2000 by CHF, Inc.


Self-connected 3D architecture of microwires.
A defect or disclination line, prepared at a designed place in a nematic liquid crystal, is used as a template for realizing a microwire directly connected to the electrodes with an accuracy of a few microm. The line attracts and traps silica particles, until self-assembling a complete micronecklace. We then fix the colloids in the necklace by means of pyrrole electropolymerization. The process may be extended to connect many microwires simultaneously. This provides a new route for manufacturing automatic 3D connections, which could be important for electronic applications as Moore's law now seems to reach some limit in 2D.


Metal ion effects on ion channel gating.
Metal ions affect ion channels either by blocking the current or by modifying the gating. In the present review we analyse the effects on the gating of voltage-gated channels. We show that the effects can be understood in terms of three main mechanisms. Mechanism A assumes screening of fixed surface charges. Mechanism B assumes binding to fixed charges and an associated electrostatic modification of the voltage sensor. Mechanism C assumes binding and an associated non electrostatic modification of the gating. To quantify the non-electrostatic effect we introduced a slowing factor, A. A fourth mechanism (D) is binding to the pore with a consequent pore block, and could be a special case of Mechanisms B or C. A further classification considers whether the metal ion affects a single site or multiple sites. Analysing the properties of these mechanisms and the vast number of studies of metal ion effects on different voltage-gated on channels we conclude that group 2 ions mainly affect channels by classical screening (a version of Mechanism A). The transition metals and the Zn group ions mainly bind to the channel and electrostatically modify the gating (Mechanism B), causing larger shifts of the steady-state parameters than the group 2 ions, but also different shifts of activation and deactivation curves. The lanthanides mainly bind to the channel and both electrostatically and non-electrostatically modify the gating (Mechanisms B and C). With the exception of the ether-à-go-go-like channels, most channel types show remarkably similar ion-specific sensitivities.


Biomimetic cardiac microsystems for pathophysiological studies and drug screens.
Microfabricated organs-on-chips consist of tissue-engineered 3D in vitro models, which rely on engineering design and provide the physiological context of human organs. Recently, significant effort has been devoted to the creation of a biomimetic cardiac system by using microfabrication techniques. By applying allometric scaling laws, microengineered cardiac systems simulating arterial flow, pulse properties, and architectural environments have been implemented, allowing high-throughput pathophysiological experiments and drug screens. In this review, we illustrate the recent trends in cardiac microsystems with emphasis on cardiac pumping and valving functions. We report problems and solutions brought to light by existing organs-on-chip models and discuss future directions of the field. We also describe the needs and desired design features that will enable the control of mechanical, electrical, and chemical environments to generate functional in vitro cardiac disease models.


Neofunctionalization of a Duplicate dachshund Gene Underlies the Evolution of a Novel Leg Segment in Arachnids.
The acquisition of a novel function, or neofunctionalization, protects duplicated genes from redundancy and subsequent loss, and is a major force that drives adaptive evolution. Neofunctionalization has been inferred for many duplicated genes based on differences in regulation between the parental gene and its duplicate. However, only few studies actually link the new function of a duplicated gene to a novel morphological or physiological character of the organism. Here we show that the duplication of dachshund (dac) in arachnids (spiders and allies) is linked with the evolution of a novel leg segment, the patella. We have studied dac genes in two distantly related spider species, the entelegyne spider Parasteatoda tepidariorum and the haplogyne spider Pholcus phalangioides. Both species possess two paralogous dac genes that duplicated before the split between entelegyne and haplogyne spiders. In contrast to the evolutionarily highly conserved dac1, its duplicate dac2 is strongly expressed in the patella leg segment during embryogenesis in both species. Using parental RNA interference in P. tepidariorum we show that dac2 is required for the development of the patella segment. If dac2 function is impaired, then the patella is fused with the tibia into a single leg segment. Thus, removing the function of dac2 experimentally reverts P. tepidariorum leg morphology into a stage before the duplication of dac and the evolution of the patella segment. Our results indicate that the origin of the patella is the result of the duplication and subsequent neofunctionalization of dac in the arachnid lineage.


Clinical and epidemiological aspects of cases of tuberculosis associated with diabetes in Salvador, Bahia, Brazil.
This study compares the clinical and epidemiological features of patients with TB, with and without DM. New cases of active pulmonary TB that occurred in Salvador, Bahia, Brazil between 2008 and 2010 were included. DM was diagnosed by based on diagnostic criteria established by the American Diabetes Association. S: Of the 323 cases of TB, 44 (13.6%) were diabetic . Patients with TB and DM were older (44.7%), with a high level of education (34.%); had slow wound healing (23.4%) and fatigue (61.7%). Clinical symptoms differed between patients with TB, with and without DM.


Prognostic significance of pleural/pericardial effusion and treatment optimization of PMBL.
The optimal treatment strategy for primary mediastinal large B-cell lymphoma (PMBL) remains unknown. We retrospectively analyzed 345 patients with newly diagnosed PMBL to identify prognostic factors and optimal treatments. Focusing on patients treated with cyclophosphamide, doxorubicin, vincristine, prednisolone, and rituximab (R-CHOP) (N=187), a higher International Prognostic Index (IPI) and the presence of pleural or pericardial effusion were identified as adverse prognostic factors for overall survival (OS) in patients treated with R-CHOP without consolidative radiation therapy (RT) [IPI: hazard ratio (HR), 4.23; 95% confidence interval (CI), 1.48-12.13; P=0.007; effusion: HR, 4.93; 95% CI, 1.37-17.69; P=0.015]. Combined with IPI and the presence of pleural or pericardial effusion for the stratification of patients treated with R-CHOP without RT, those with lower IPI and the absence of effusion comprised approximately onehalf of these patients and could be identified as curable [OS and progression free survival (PFS) at 4 years, 95% and 87%, respectively)] Taken together, our simple indicators of IPI and the presence of effusion could stratify patients with PMBL and thereby facilitate treatment selection.


Ligamentous balancing in rotating-platform knees.
The primary objectives of total knee arthroplasty are to reduce pain, maximize range of motion (ROM), and provide stability through the gait cycle. Pain relief depends on proper ligamentous balancing and solid implantation. Solid implantation is typically assured through cementation, whereas soft-tissue balancing is vital to ensure maximum ROM and stability. Special consideration should be given to obtaining proper axial alignment, symmetric tension on the ligaments in both flexion and extension, and equalizing the flexion extension gaps. Strategies to ensure proper alignment and ligamentous balancing intraoperatively are discussed.


Is lower uterine segment involvement a prognostic factor in endometrial cancer?
The purpose of this study is to investigate the prognostic significance of lower uterine segment (LUS) involvement in endometrial cancer (EC). We reviewed the patients who were operated at our institution between July 2007 and March 2015 with the diagnosis of EC. Tumors localized in the corpus and involving the LUS or localized entirely in the LUS formed Group A, while tumors in the uterine corpus without LUS involvement formed Group B. Clinicopathological characteristics and survival of the patients were compared in both groups. A total of 500 patients were included in the study. There were 139 patients who had tumors involving the LUS and formed Group A, while 361 patients with endometrial tumors in the uterine corpus without LUS involvement formed Group B. We did not detect a significant difference between survival of the patients in group A and group B (78 months vs. 87 months, respectively; P > 0.05). We found that LUS involvement was not an independent prognostic factor for poor survival, but it is associated with other poor prognostic factors such as deep myometrial invasion, uterine serosal involvement, lymphovascular space invasion, lymph node metastasis and higher FIGO grade.


Vitamin D and hippocampal development-the story so far.
Epidemiological studies suggest that vitamin D insufficiency may be prevalent in young as well as older populations. The pleiotropic effects of vitamin D are now beyond dispute and a growing number of studies provide accumulating evidence of a role for vitamin D in brain development and function. A number of studies to date have investigated the effects of early-life vitamin D deprivation on adult hippocampus in animals and humans, and there is a growing body of evidence to suggest a role for this hormone in the development of selected hippocampal functions such as latent inhibition and hole board habituation in rats. There are few studies to date of vitamin D deprivation or supplementation on early hippocampal development in vivo. However, a small number of studies, mostly in vitro, point to a role for vitamin D in differentiation and development of hippocampal neurons. There is also limited evidence that supplementation with vitamin D following a period of deprivation is capable of restoring cellular activity and later function. Further avenues of future research are outlined including animal studies on the effects of vitamin D deprivation and inadequacy on early hippocampal biochemistry and function, e.g., measurement of BDNF levels, GABAergic activity, long-term potentiation (LTP) and spatial navigation. It also remains to be established if there are critical developmental windows during which vitamin D is required. In light of the importance of the hippocampus in LTP and spatial learning, further investigations on the early effects of vitamin D deprivation on hippocampal development are warranted.


Adaptive correction of craniofacial defects in pre-metamorphic Xenopus laevis tadpoles involves thyroid hormone-independent tissue remodeling.
Although it is well established that some organisms can regenerate lost structures, the ability to remodel existing malformed structures has been less well studied. Therefore, in this study we examined the ability of pre-metamorphic Xenopus laevis tadpoles to self-correct malformed craniofacial tissues. We found that tadpoles can adaptively improve and normalize abnormal craniofacial morphology caused by numerous developmental perturbations. We then investigated the tissue-level and molecular mechanisms that mediate the self-correction of craniofacial defects in pre-metamorphic X. laevis tadpoles. Our studies revealed that this adaptive response involves morphological changes and the remodeling of cartilage tissue, prior to metamorphosis. RT-qPCR and RNA-seq analysis of gene expression suggests a thyroid hormone-independent endocrine signaling pathway as the potential mechanism responsible for triggering the adaptive and corrective remodeling response in these larvae that involves mmp1 and mmp13 upregulation. Thus, investigating how malformed craniofacial tissues are naturally corrected in X. laevis tadpoles has provided valuable insights into the maintenance and manipulation of craniofacial morphology in a vertebrate system. These insights may help in the development of novel therapies for developmental craniofacial anomalies in humans.


Investigating the Relationship between Structural Features of Built Environment and Physical Activity using Geographic Information Systems (GIS).
Built environment determinants of engaging in physical activity are dependent on the properties of societies. This study investigates the relationship between structural variables of the environment using geographic information systems (GIS) and the level of physical activity in 22 districts of Tehran. This cross-sectional study was based on Urban Health Equity Assessment and Response Tool (Urban HEART-2). Physical activity level was assessed via the Global Physical Activity Questionnaire (GPAQ). The characteristics of the neighborhood environment, including land use, street pattern, population density, and traffic, were determined via ArcGIS software. Walkability index (population density, street pattern, land use) was calculated to assess the effect of the main variables of living environment on physical activity level. Among the built environmental variables, land use was associated with the total physical activity and travel-related physical activity level (r: 0.155, P value: 0.001, and r: 0.122, P value: 0.007, respectively). The walkability index indicated an association with the total physical activity level and travel-related physical activity level (r: 0.126, P value: 0.006, and r: 0.135, P value: 0.001, respectively). Higher levels of the walkability index were associated with an improved level of physical activity (OR: 2.04). Walkability index and land use positively correlate with total physical activity level, and providing action plans that improve walkability index and land use might lead to increased physical activity level.


The zinc binding site of the Shaker channel KDC1 from Daucus carota.
KDC1 is a voltage-dependent Shaker-like potassium channel subunit cloned from Daucus carota which produces conductive channels in Xenopus oocytes only when coexpressed with other plant Shaker potassium subunits, such as KAT1 from Arabidopsis thaliana. External Zn(2+) determines a potentiation of the current mediated by the dimeric construct KDC1-KAT1, which has been ascribed to zinc binding at a site comprising three histidines located at the S3-S4 (H161, H162) and S5-S6 (H224) linkers of KDC1. Here we demonstrate that also glutamate 164, located in close proximity of the KDC1 S4 segment, is an essential component of the zinc-binding site. On the contrary, glutamate 159, located in symmetrical position with respect to E164 in the sequence E(159)XHHXE(164) but more distant from the voltage sensor, does not play any role in zinc binding. The effects of Zn(2+) can be expressed as a "shift" of the gating parameters along the voltage axis. Kinetic modeling shows that Zn(2+) slows the closing kinetics of KDC1-KAT1 without affecting the opening kinetics. Possibly, zinc affects the movement of the voltage sensor in and out of the membrane phase through electrostatic modification of a site close to the voltage sensor.


Leishmania-Specific Promiscuous Membrane Protein Tubulin Folding Cofactor D Divulges Th1/Th2 Polarization in the Host via ERK-1/2 and p38 MAPK Signaling Cascade.
Visceral leishmaniasis (VL)-related mortality and morbidity imposes a great deal of health concern across the globe. The existing anti-leishmanial drug regimen generally fails to eliminate newly emerging resistant isolates of this dreadful parasite. In such circumstances, the development of a prophylactic strategy to impart protection against the disease is likely to take center stage. In order to develop a promising prophylactic vaccine, it is desirable to identify an adequately potential vaccine candidate. In silico analysis of Leishmania tubulin folding cofactor D protein predicted its potential to activate both B- and T-cell repertoires. Furthermore, the ELISA employing anti-peptide27 (a segment of tubulin folding cofactor D) antibody revealed its proficiency in VL diagnosis and treatment monitoring. The peptide27 and its cocktail with another Leishmania peptide (peptide23) prompted the up-regulation of pro-inflammatory cytokines, such as IFN-γ, TNF-α, IL-2, IL-17, etc., and the down-regulation of immune-regulatory cytokines, such as IL-10, in the immunized BALB/c mice. Coherent to the consequence of peptide-specific humoral immune response, peptide cocktail-based immunization ensued in the predominant amplification of pathogen-specific IgG2a over the IgG1 isotype, up-regulated proliferation of T lymphocytes, and enhanced production of nitric oxide, reactive oxygen species, etc. We also established that the peptide cocktail modulated host MAPK signaling to favor the amplification of Th1-dominated immune response in the host. The peptide cocktail mediated the activation of the host immune armory, which was eventually translated into a significant decline in parasitic load in the visceral organs of experimental animals challenged with Leishmania donovani.


Secondary Chondrosarcoma from an Osteochondroma of the Proximal Tibia Involving the Fibula.
There are few reports on the surgical treatment of secondary malignancy arising from an osteochondroma on the lateral side of the proximal tibia. From March 2008 to December 2011, 3 patients were treated for a secondary chondrosarcoma from an osteochondroma of the proximal tibia involving the fibula. The operative procedure can be summed up as follows: (1) resection of the tumor including the fibula; (2) preservation of the peroneal nerve and the fibular head; and (3) arthrodesis of the proximal tibiofibular joint. Serial radiological studies showed successful fusion in the proximal tibiofibular joint in all patients. The Musculoskeletal Tumor Society functional scores were excellent in all 3 patients. No patients showed instability of the ipsilateral knee joint in any direction. All 3 patients could return to sports activities. Until the last follow-up, there was no evidence of disease recurrence. We suggest that the operative procedure described in this article would provide satisfactory oncological and functional outcomes.


Apolipoprotein E structural requirements for the formation of SDS-stable complexes with beta-amyloid-(1-40): the role of salt bridges.
Of the three major isoforms of human apolipoprotein E (apoE), apoE4 is a risk factor for the development of Alzheimer's disease. Among possible neurologically relevant differences in the properties of apoE3 and apoE4 is the fact that apoE3 forms an SDS-stable complex with beta-amyloid-(1-40) (Abeta40) with greater avidity than does apoE4. This interaction may sequester potentially toxic species of Abeta or facilitate clearance. To understand more about this difference, we examined whether differences in salt bridges between apoE domains influence the capacity of apoE isoforms to form complexes with Abeta. In apoE3 there is a salt bridge between Arg-61 and Asp-65, while in apoE4 there are salt bridges between Arg-61 and Glu-255, and Arg-112 and Glu-109. Mutation of position 112, which is Cys in apoE3 and Arg in apoE4, to Ala or Lys abolished complex formation, while mutant apoE with Ser at this position retained the capacity to form complex. Substituting Ala for Glu-109 had no effect on the ability of either apoE4 or apoE3 to form complexes. On the other hand, substitution of Thr for Arg-61 in apoE3 abolished, and truncation of apoE3 at position 201 substantially lowered, but did not abolish, complex formation. Neither of these mutations within apoE4 had any affect on its complex formation with Abeta. These results suggest that the nature of the cysteine residue in apoE3 and interactions between the N-terminal and C-terminal domains of human apoE are important for the ability of apoE3 to form an SDS-stable complex with Abeta40.


From gene to dose: Long-read sequencing and *-allele tools to refine phenotype predictions of CYP2C19.
Background: Inter-individual differences in drug response based on genetic variations can lead to drug toxicity and treatment inefficacy. A large part of this variability is caused by genetic variants in pharmacogenes. Unfortunately, the Single Nucleotide Variant arrays currently used in clinical pharmacogenomic (PGx) testing are unable to detect all genetic variability in these genes. Long-read sequencing, on the other hand, has been shown to be able to resolve complex (pharmaco) genes. In this study we aimed to assess the value of long-read sequencing for research and clinical PGx focusing on the important and highly polymorphic CYP2C19 gene. Methods and Results: With a capture-based long-read sequencing panel we were able to characterize the entire region and assign variants to their allele of origin (phasing), resulting in the identification of 813 unique variants in 37 samples. To assess the clinical utility of this data we have compared the performance of three different *-allele tools (Aldy, PharmCat and PharmaKU) which are specifically designed to assign haplotypes to pharmacogenes based on all input variants. Conclusion: We conclude that long-read sequencing can improve our ability to characterize the CYP2C19 locus, help to identify novel haplotypes and that *-allele tools are a useful asset in phenotype prediction. Ultimately, this approach could help to better predict an individual's drug response and improve therapy outcomes. However, the added value in clinical PGx might currently be limited.


Outer Membrane Permeability of Cyanobacterium Synechocystis sp. Strain PCC 6803: Studies of Passive Diffusion of Small Organic Nutrients Reveal the Absence of Classical Porins and Intrinsically Low Permeability.
The outer membrane of heterotrophic Gram-negative bacteria plays the role of a selective permeability barrier that prevents the influx of toxic compounds while allowing the nonspecific passage of small hydrophilic nutrients through porin channels. Compared with heterotrophic Gram-negative bacteria, the outer membrane properties of cyanobacteria, which are Gram-negative photoautotrophs, are not clearly understood. In this study, using small carbohydrates, amino acids, and inorganic ions as permeation probes, we determined the outer membrane permeability of Synechocystis sp. strain PCC 6803 in intact cells and in proteoliposomes reconstituted with outer membrane proteins. The permeability of this cyanobacterium was >20-fold lower than that of Escherichia coli The predominant outer membrane proteins Slr1841, Slr1908, and Slr0042 were not permeable to organic nutrients and allowed only the passage of inorganic ions. Only the less abundant outer membrane protein Slr1270, a homolog of the E. coli export channel TolC, was permeable to organic solutes. The activity of Slr1270 as a channel was verified in a recombinant Slr1270-producing E. coli outer membrane. The lack of putative porins and the low outer membrane permeability appear to suit the cyanobacterial autotrophic lifestyle; the highly impermeable outer membrane would be advantageous to cellular survival by protecting the cell from toxic compounds, especially when the cellular physiology is not dependent on the uptake of organic nutrients.IMPORTANCE Because the outer membrane of Gram-negative bacteria affects the flux rates for various substances into and out of the cell, its permeability is closely associated with cellular physiology. The outer membrane properties of cyanobacteria, which are photoautotrophic Gram-negative bacteria, are not clearly understood. Here, we examined the outer membrane of Synechocystis sp. strain PCC 6803. We revealed that it is relatively permeable to inorganic ions but is markedly less permeable to organic nutrients, with >20-fold lower permeability than the outer membrane of Escherichia coli Such permeability appears to fit the cyanobacterial lifestyle, in which the diffusion pathway for inorganic solutes may suffice to sustain the autotrophic physiology, illustrating a link between outer membrane permeability and the cellular lifestyle.


Hyperthyroidism--a disease of old age?
In a three-year project in which thyroid studies were undertaken for broad indications 49 cases of thyrotoxicosis were diagnosed; 28 of these patients were 60 years of age or older. Sixteen of these older patients had different forms of atypical thyrotoxicosis, and most were completely without the usual symptoms of the condition. It is concluded that thyrotoxicosis is more frequent in persons over 60 than in younger persons. Many older patients have various forms of masked thyrotoxicosis, and in many cases the correct diagnosis can be made only by undertaking thyroid studies on very wide indications.


Inhibition of nuclear factor-kappa B enhances the tumor growth of ovarian cancer cell line derived from a low-grade papillary serous carcinoma in p53-independent pathway.
NF-kB can function as an oncogene or tumor suppressor depending on cancer types. The role of NF-kB in low-grade serous ovarian cancer, however, has never been tested. We sought to elucidate the function of NF-kB in the low-grade serous ovarian cancer. The ovarian cancer cell line, HOC-7, derived from a low-grade papillary serous carcinoma. Introduction of a dominant negative mutant, IkBαM, which resulted in decrease of NF-kB function in ovarian cancer cell lines. The transcription ability, tumorigenesis, cell proliferation and apoptosis were observed in derivative cell lines in comparison with parental cells. Western blot analysis indicated increased expression of the anti-apoptotic proteins Bcl-xL and reduced expression of the pro-apoptotic proteins Bax, Bad, and Bid in HOC-7/IĸBαM cell. Further investigations validate this conclusion in KRAS wildtype cell line SKOV3. Interesting, NF-kB can exert its pro-apoptotic effect by activating mitogen-activated protein kinase (MAPK) phosphorylation in SKOV3 ovarian cancer cell, whereas opposite changes detected in p-MEK in HOC-7 ovarian cancer cell, the same as some chemoresistant ovarian cancer cell lines. In vivo animal assay performed on BALB/athymic mice showed that injection of HOC-7 induced subcutaneous tumor growth, which was completely regressed within 7 weeks. In comparison, HOC-7/IĸBαM cells caused sustained tumor growth and abrogated tumor regression, suggesting that knock-down of NF-kB by IĸBαM promoted sustained tumor growth and delayed tumor regression in HOC-7 cells. Our results demonstrated that NF-kB may function as a tumor suppressor by facilitating regression of low grade ovarian serous carcinoma through activating pro-apoptotic pathways.


Study of the Size and Shape of Synapses in the Juvenile Rat Somatosensory Cortex with 3D Electron Microscopy.
Changes in the size of the synaptic junction are thought to have significant functional consequences. We used focused ion beam milling and scanning electron microscopy (FIB/SEM) to obtain stacks of serial sections from the six layers of the rat somatosensory cortex. We have segmented in 3D a large number of synapses (n = 6891) to analyze the size and shape of excitatory (asymmetric) and inhibitory (symmetric) synapses, using dedicated software. This study provided three main findings. Firstly, the mean synaptic sizes were smaller for asymmetric than for symmetric synapses in all cortical layers. In all cases, synaptic junction sizes followed a log-normal distribution. Secondly, most cortical synapses had disc-shaped postsynaptic densities (PSDs; 93%). A few were perforated (4.5%), while a smaller proportion (2.5%) showed a tortuous horseshoe-shaped perimeter. Thirdly, the curvature was larger for symmetric than for asymmetric synapses in all layers. However, there was no correlation between synaptic area and curvature.


[Malformed pygopagus conjoined twins in a spontaneous triplet pregnancy].
This is a case study of malformed conjoined twins in a spontaneous triplet pregnancy. A 20-year-old primigravida spontaneously delivered at 22 weeks gestation a normal 425-gram male newborn followed by a delivery of a 525 grams pygopagus conjoined male twins. One twin had omphalocele, missing ears, cleft palate and dysmorphic figures while the other had imperforated anus and dysmorphic figures. The Apgar scores were 2/0. The placenta was monochorionic/diamniotic with single umbilical artery supplying both conjoined twins. Conjoined twin in a triplet pregnancy may be diagnosed by ultrasound as early as the first trimester. Failure in diagnosis of the condition resulted in early delivery of three premature infants. All three infants died after delivery.


[Complications of intramedullary nailing and their prevention].
Interlocking nailing technique offers a wide variety of indications which are associated with an increased risk of complications. A consecutive series of 208 cases were analyzed and possible complications, how to avoid them and recommendations for treatment are given. The complications were divided into common complications--like infection, pseudarthrosis, malrotation, compartment-syndrome, disorders of the arterial and venous system, neurological deficts and fat embolism--as well as into specific complications--like malreduction of the fracture, malpositioning of the guide wire, splitting fragments and problems with nail insertion and with the locking bolts. There was a 8.7% common complication rate. Specific complications occurred intraoperatively in 3.5% and postoperatively in 11% of call cases. This seems to be a high rate which is decreasing by progressive experience of the surgeon. In many cases of shaft fractures interlocking nailing technique is the method of choice.


Redescription of Yphthimoides patricia (Hayward, 1957), with taxonomic notes on the names Euptychia saltuensis Hayward, 1962 and Yphthimoides manasses (C. Felder R. Felder, 1867) (Nymphalidae: Satyrinae).
Euptychia saltuensis Hayward, 1962, new synonym, currently regarded as a nomen dubium and possibly a junior subjective synonym of Yphthimoides manasses (C. Felder R. Felder, 1867), is here treated as a junior subjective synonym of Yphthimoides patricia (Hayward, 1957), based on morphological characters of the male genitalia and the DNA barcode. The taxonomic status of Y. patricia is re-examined, and a detailed redescription of the adult morphology, including the male genitalia, is presented. Information on the distribution, habitat and immature stages of Y. patricia is also provided. Yphthimoides patricia is clearly a distinct species from Y. manasses based on the analysis of DNA barcode sequences and the morphology of the male genitalia.


Growth performance and feed utilization of keureling fish Tor tambra (Cyprinidae) fed formulated diet supplemented with enhanced probiotic.
Background The objective of the present study was to determine the optimum dosage of probiotic in the diet of keureling fish ( Tor tambra) fry. MethodsLactobacillus casei from Yakult® was used as a starter, and enhanced with Curcuma xanthorrhiza, Kaempferia galanga and molasses. The mixture was fermented for 7 days prior to use as probiotic in a formulated diet containing 30% crude protein. Four levels of probiotic dosage; 0 ml kg -1 (control), 5 ml kg -1, 10 ml kg -1 and 15 ml kg -1 were tested in this study. The fish was fed twice a day at 08.00 AM and 06.00 PM at the ration of 5% body weight for 80 days. Results The results showed that growth performance and feed efficiency increased with increasing probiotic dosage in the diet from control (no probiotic) to 10 ml kg -1 of probiotic dosage and then decreased when the dosage was increased up to 15 ml kg -1. Conclusions The best values for all measured parameters were recorded at the dosage of 10 ml kg -1. Therefore, it was concluded that the optimum dosage of enhanced probiotic for T. tambra fry was 10 ml kg -1 of feed.


Predicting Adverse Perioperative Events in Patients Undergoing Primary Cleft Palate Repair.
This study aimed to identify risk factors for adverse perioperative events (APEs) after cleft palatoplasty to develop an individualized risk assessment tool. Retrospective cohort. Tertiary institutional. Patients younger than 2 years with cleft palate. Primary Furlow palatoplasty between 2008 and 2011. Adverse perioperative event, defined as laryngo- or bronchospasm, accidental extubation, reintubation, obstruction, hypoxia, or unplanned intensive care unit admission. Three hundred patients averaging 12.3 months old were included. Cleft distribution included submucous, 1%; Veau 1, 17.3%; Veau 2, 38.3%; Veau 3, 30.3%; and Veau 4, 13.0%. Pierre Robin (n = 43) was the most prevalent syndrome/anomaly. Eighty-three percent of patients received reversal of neuromuscular blockade, and total morphine equivalent narcotic dose averaged 0.19 mg/kg. Sixty-nine patients (23.0%) had an APE, most commonly hypoventilation (10%) and airway obstruction (8%). Other APEs included reintubation (4.7%) and laryngobronchospasm (3.3%). APE was associated with multiple intubation attempts (odds ratio [OR] = 6.6, P = .001), structural or functional airway anomaly (OR = 4.5, P < .001), operation >160 minutes (OR = 2.2, P = .04), narcotic dose >0.3 mg/kg (OR = 2.3, P = .03), inexperienced provider (OR = 2.1, P = .02), and no paralytic reversal administration (OR = 2.0, P = .049); weight between 9 and 13 kg was protective (OR = 0.5, P = .04). Patients were risk-stratified according to individual profiles as low, average, high, or extreme risk (APE 2.5%-91.7%) with excellent risk discrimination (C-statistic = 0.79). APE incidence was 23.0% after palatoplasty, with a 37-fold higher incidence in extreme-risk patients. Individualized risk assessment tools may enhance perioperative clinical decision making to mitigate complications.


High-pressure liquid chromatographic analysis of ceftazidime in serum and urine.
A rapid and sensitive high-pressure liquid chromatographic procedure was developed for quantitative analysis of a new semisynthetic cephalosporin, ceftazidime, in serum and urine. A good linear relationship was established between peak height and the amount of ceftazidime injected over a concentration range of 1.9 to 30 micrograms/ml. Recovery was approximately 90% at concentrations of 3, 15, and 30 micrograms/ml. The method is specific for ceftazidime, with no interference noted from 11 other beta-lactam antibiotics tested. The assay is accurate, reproducible, and useful for pharmacokinetic studies in humans as demonstrated in two subjects.


Freeze-fracture replica studies of effects of ACTH treatment and hypophysectomy on the cell surface of the rat adrenal inner cortex.
The morphological change of the cell surface in the inner zone of the adrenal cortex was studied in intact, hypophysectomized, and ACTH-treated hypophysectomized rats, using freeze-fracture replicas. In both intact and hypophysectomized ACTH-treated animals, the P fracture face of the plasma membranes of the cortical cells facing capillaries shows groups of characteristic finger-shaped microvilli, which arise horizontally from the edges of a small cavity on the cell surface to form various configurations ranging from incompletely organized, flower bud-like to complicated, arboraceous ones. In the non-treated hypophysectomized animals, however, the microvilli in such groups decrease in number and shorten in length but increase in width, looking like a blade of a prickly-pear cactus or an irregular-shaped plate. On the other hand, presumptive exocytotic and endocytotic specializations of intra-membraneous and experimental animals.


Controls that expedite first-passage times in disordered systems.
First-passage time statistics in disordered systems exhibiting scale invariance are studied widely. In particular, long trapping times in energy or entropic traps are fat-tailed distributed, which slow the overall transport process. We study the statistical properties of the first-passage time of biased processes in different models, and we employ the big-jump principle that shows the dominance of the maximum trapping time on the first-passage time. We demonstrate that the removal of this maximum significantly expedites transport. As the disorder increases, the system enters a phase where the removal shows a dramatic effect. Our results show how we may speed up transport in strongly disordered systems exploiting scale invariance. In contrast to the disordered systems studied here, the removal principle has essentially no effect in homogeneous systems; this indicates that improving the conductance of a poorly conducting system is, theoretically, relatively easy as compared to a homogeneous system.


Targeted disruption of the neurofibromatosis type-1 gene leads to developmental abnormalities in heart and various neural crest-derived tissues.
The neurofibromatosis (NF1) gene shows significant homology to mammalian GAP and is an important regulator of the ras signal transduction pathway. To study the function of NF1 in normal development and to try and develop a mouse model of NF1 disease, we have used gene targeting in ES cells to generate mice carrying a null mutation at the mouse Nf1 locus. Although heterozygous mutant mice, aged up to 10 months, likely attributable to a severe malformation of the heart. Interestingly, mutant embryos also display hyperplasia of neural crest-derived sympathetic ganglia. These results identify new roles for NF1 in development and indicate that some of the abnormal growth phenomena observed in NF1 patients can be recapitulated in neurofibromin-deficient mice.


Hydration and energy dissipation measurements of biomolecules on a piezoelectric quartz oscillator by admittance analyses.
By using a 27-MHz piezoelectric quartz oscillator connected with a vector network analyzer, we obtained resonance frequency decreases (-DeltaFwater) and energy dissipation increases (DeltaDwater) during binding of biotinylated bovine serum albumin, biotinylated ssDNA, biotinylated dsDNA, and biotinylated pullulan to a NeutrAvidin-immobilized 27-MHz quartz crystal microbalance (QCM) plate in aqueous solution, as well as in the wet air phase (98% humidity, -DeltaFwet and DeltaDwet) and in the dry air phase (-DeltaFair and DeltaDair). -DeltaFwater indicates the total mass of the molecule, bound water, and vibrated water in aqueous solutions. -DeltaFwet indicates the total mass of the molecule and bound water. -DeltaFair simply shows the real mass of the molecule on the QCM. In terms of results, (-DeltaFwet)/(-DeltaFair) values indicated the bound water ratios per unit biomolecular mass were on the order of pullulan (2.1-2.2) > DNAs = proteins (1.4-1.6) > polystyrene (1.0). The (-DeltaFwater)/(-DeltaFair) values indicated the hydrodynamic water (bound and vibrated water) ratios per unit biomolecular mass were on the order of dsDNA (6.5) > ssDNA = pullulan (3.5-4.4) > proteins (2.4-2.5) > polystyrene (1.0). Energy dissipation parameters per unit mass in water (DeltaDwater/(-DeltaFair)) were on the order of pullulan > dsDNA > ssDNA > proteins > polystyrene. Energy dissipation in the wet and dry air phases (DeltaDwet and DeltaDair) were negligibly small, which indicates even these biomolecules act as elastic membranes in the air phase (without aqueous solution). We obtained a good linear relationship between [(-DeltaFwater)/(-DeltaFair) - 1], which is indicative of hydration and DeltaDwater/(-DeltaFair) of proteins. The aforementioned values suggest that the energy dissipation of proteins was mainly caused by hydration and that proteins themselves are elastic molecules without energy dissipation in aqueous solutions. On the contrary, plots in cases of denatured proteins, DNAs, and pullulans were relatively deviant toward the large hydration and energy dissipation from the theoretical line as perfect elastic materials, meaning that the large energy dissipation occurs because of viscoelastic properties of denatured proteins, linear DNAs, and pullulans in the water phase, in addition to energy dissipation due to the hydration of molecules. These two parameters could characterize various biomolecules with structural properties in aqueous solutions.


Amyloidogenic behavior of different intermediate state of stem bromelain: A biophysical insight.
Stem bromelain, a cysteine proteases from Ananas comosus is a widely accepted therapeutic drug with broad medicinal application. It exists as intermediate states at pH 2.0 and 10.0, where it encountered in gastrointestinal tract during adsorption (acidic pH) and in gut epithelium (alkaline pH), respectively. In this study, we monitored the thermal aggregation/amyloid formation of SB at different pH intermediate states. Thermal treatment of stem bromelain at pH 10.0 favors the fibrillation in which the extent of aggregation increases with increase in protein concentration. However, no fibril formation in stem bromelain at pH 2.0 was found at all the concentration used at pH 10.0. The fibril formation was confirmed by various techniques such as turbidity measurements, Rayleigh light scattering, dye binding assays and far UV circular dichroism. The Dynamic light scattering confirmed the formation of aggregates by measuring the hydrodynamic radii pattern. Moreover, microscopic techniques were performed to analyze the morphology of fibrils. The aggregation behavior may be due to variation in number of charged amino acid residues. The less negative charge developed at pH 10.0 may be responsible for aggregation. This work helps to overcome the aggregation related problems of stem bromelain during formulations in pharmaceutical industry.


Stereoselective total synthesis of (-)-nupharamine utilizing an α-chlorosulfide and a sulfinimine for C-C bond formation.
An efficient stereoselective synthesis of the nuphar alkaloid, (-)-nupharamine, is reported. The key features include the Lewis acid catalyzed reaction of an α-chlorosulfide with a silyl ketene acetal for C-C bond formation, creation of the stereocenter at C2 by a diastereoselective reaction of allyl indium with a sulfinimine and reductive amination for the introduction of the C6 stereocenter of the piperidine ring.


Detection and characterization of lung cancer using cell-free DNA fragmentomes.
Non-invasive approaches for cell-free DNA (cfDNA) assessment provide an opportunity for cancer detection and intervention. Here, we use a machine learning model for detecting tumor-derived cfDNA through genome-wide analyses of cfDNA fragmentation in a prospective study of 365 individuals at risk for lung cancer. We validate the cancer detection model using an independent cohort of 385 non-cancer individuals and 46 lung cancer patients. Combining fragmentation features, clinical risk factors, and CEA levels, followed by CT imaging, detected 94% of patients with cancer across stages and subtypes, including 91% of stage I/II and 96% of stage III/IV, at 80% specificity. Genome-wide fragmentation profiles across ~13,000 ASCL1 transcription factor binding sites distinguished individuals with small cell lung cancer from those with non-small cell lung cancer with high accuracy (AUC = 0.98). A higher fragmentation score represented an independent prognostic indicator of survival. This approach provides a facile avenue for non-invasive detection of lung cancer.


Evidence for adaptive evolution at the divergence between lymphoid and brain HIV-1 nef genes.
Human immunodeficiency virus type 1 (HIV) infection of the central nervous system frequently causes HIV-associated neurocognitive disorders (HAND). The role of HIV Nef and other accessory proteins in HAND pathogenesis is unclear. To determine whether HIV nef undergoes adaptive selection in brain, we cloned 100 nef sequences (n = 30 brain and n = 70 lymphoid) from four patients with AIDS and HIV-associated dementia (HAD). Normalized nonsynonymous substitutions were more frequent at the divergence of lymphoid and brain sequences, indicating stronger adaptive selection in brain compared to lymphoid tissue. Brain-specific nonsynonymous substitutions were found within an NH(3)-terminal CTL epitope, the PACS1 binding motif, or positions predicted to be important for activation of the myeloid-restricted Src family tyrosine kinase Hck. These results suggest that adaptive selection of HIV nef in brain may reflect altered requirements for efficient replication in macrophages and brain-specific immune selection pressures.


All India Ophthalmological Society - Preferred practice in refractive surgery during the COVID-19 pandemic.
Coronavirus pandemic has strained the healthcare system with mortality and morbidity. A number of elective surgeries have come to standstill due to lockdown and movement restrictions. Refractive surgery being a purely elective procedure and quite a fresh subset of ophthalmology, there is a lack of unanimity as to what precautions should be followed to resume the practice of same. This article attempts to highlight simple guidelines in accordance with an expert panel, which can be followed by all those involved directly or indirectly in refractive surgery services while addressing safety of doctors, supporting staff as well as patients as a primary concern.


Acid phosphatase activity in mouse brain infected with Venezuelan equine encephalomyelitis virus.
The mode of development of Venezuelan equine encephalomyelitis virus and the activity of acid phosphatase in the central nervous system of newborn mice were investigated. Precursor particles appeared to be formed in masses of viroplasm, migrating to the membrane of the Golgi cisterns and vacuoles or to the plasma membrane and being transformed into mature viral particles by budding. Mature viral particles were also found in the lumen of the blood vessels and around the myelin sheath of axons. Increased number of Golgi complexes and depletion of polysomes were the main ultrastructural alterations of the nerve cells. Acid phosphatase activity was found to be increased in the Golgi cisterns, vacuoles, and lysosomes of nerve cells. The presence of acid phosphatase activity in the rough endoplasmic reticulum and perinuclear cisterns suggests increased production of the enzyme in the nerve cells infected with Venezuelan equine encephalomyelitis virus.


The risk of post-ERCP pancreatitis and the protective effect of rectal indomethacin in cases of attempted but unsuccessful prophylactic pancreatic stent placement.
It is believed, based on limited observational data, that an unsuccessful attempt to place a prophylactic pancreatic stent substantially increases the risk of post-ERCP pancreatitis (PEP). To better understand the risk of PEP in patients with failed pancreatic stent placement (FPS) and the impact of rectal indomethacin on this risk. Secondary analysis of randomized, controlled trial data. University of Michigan and Indiana University. A total of 577 clinical trial participants at elevated risk for PEP. Pancreatic stent placement. Within the placebo group, we compared PEP rates in patients with FPS, patients who underwent successful stent placement, and in those without a stent attempt. We also performed a regression analysis evaluating the association between FPS and PEP. To define the protective effect of indomethacin, we repeated these analyses in the indomethacin group and in the full study cohort. The incidence of PEP among patients in the placebo group who experienced FPS was 34.7%, significantly exceeding rates in patients who underwent successful stent placement (16.4%) and in those without a stent attempt (12.1%). After we adjusted for known PEP risk factors, FPS was found to be independently associated with PEP. Among the indomethacin group and in the full cohort, FPS was not associated with a higher risk of PEP. Low event rate, FPS not prospectively captured. FPS appears to confer an increased risk of PEP, which is attenuated by rectal indomethacin administration. These findings highlight the importance of adequate training and proficiency before endoscopists attempt pancreatic stent placement and the routine use of rectal indomethacin in high-risk ERCP cases.


Definition of regulatory network elements for T cell development by perturbation analysis with PU.1 and GATA-3.
PU.1 and GATA-3 are transcription factors that are required for development of T cell progenitors from the earliest stages. Neither one is a simple positive regulator for T lineage specification, however. When expressed at elevated levels at early stages of T cell development, each of these transcription factors blocks T cell development within a different, characteristic time window, with GATA-3 overexpression initially inhibiting at an earlier stage than PU.1. These perturbations are each associated with a distinct spectrum of changes in the regulation of genes needed for T cell development. Both transcription factors can interfere with expression of the Rag-1 and Rag-2 recombinases, while GATA-3 notably blocks PU.1 and IL-7Ralpha expression, and PU.1 reduces expression of HES-1 and c-Myb. A first-draft assembly of the regulatory targets of these two factors is presented as a provisional gene network. The target genes identified here provide insight into the basis of the effects of GATA-3 or PU.1 overexpression and into the regulatory changes that distinguish the developmental time windows for these effects.


Synthesis, structure, and magnetism of non-planar heptanuclear lanthanide(III) complexes.
The reaction of the hydrazone, 2-methoxy-6-(pyridin-2-yl-hydrazonomethyl) phenol (LH) with lanthanide(iii) nitrate salts in the presence of excess triethylamine afforded the heptanuclear Ln(iii) complexes: [Gd7(L)6(μ3-OH)8(NO3)4(H2O)]·(NO3)3·8CH3CN·H2O (), [Tb7(L)6(μ3-OH)8(NO3)4]·(NO3)3·9CH3CN·2CH3OH·3H2O (), [Dy7(L)6(μ3-OH)8(NO3)4(H2O)]·(NO3)3·7CH3CN·3H2O (), [Ho7(L)6(μ3-OH)8(NO3)4]·(NO3)3·11CH3CN·2CH3OH·2H2O () and [Er7(L)6(μ3-OH)8(NO3)4]·(NO3)3·8CH3CN·2CH3OH (). Single crystal X-ray diffraction studies reveal that these complexes are tri-cationic, possessing three nitrate counter anions. The heptanuclear ensemble is non-planar and consists of six [L](-) and eight μ3-OH ligands. These compounds show an interesting structural motif with two incomplete cubes fused to each other through a common Ln(iii) ion. Compound exhibits a magnetocaloric effect, with (-ΔSm(T) = 27.7 J kg(-1) K(-1) at 3 K and under a field change of 0-7 T), while compound shows slow magnetic relaxation at very low temperatures.


Expression of the Notch signaling pathway and effect on exocrine cell proliferation in adult rat pancreas.
When pancreatic tissue is injured after duct obstruction, acinoductal metaplasia is observed. Similar metaplastic changes occur when exocrine pancreatic cells are isolated and cultured. We demonstrate that under these experimental conditions the exocrine acinar cells lose their differentiated characteristics: expression of the acinar transcription factors p48/Ptf1alpha and Mist1 is decreased or lost, whereas expression of the embryonic transcription factor Pdx1 is increased. The receptors Notch1 and Notch2, members of the DSL family of Notch ligands, and the target genes in the Notch-signaling pathway Hes1, Hey1, and Hey2 become strongly up-regulated. We noted also reduced expression of Sel1L, a Notch repressor that is normally highly expressed in exocrine pancreas. Stimulation of Notch by its ligand Jagged1 diminished the proliferation of cultured metaplastic exocrine cells. Chemical inhibition of Notch signaling resulted in increased proliferation and induction of the cell-cycle regulator p21Cip1. This effect seems to be Hes1-independent and mainly coincides with decreased Hey1 and Hey2 mRNA expression. In conclusion, we demonstrate that during acinoductal metaplasia the Notch-signaling pathway is activated concomitantly with changes in transcription factor expression of pancreatic acinar cells. In addition, we show that Notch signaling is implicated in the suppression of proliferation of these metaplastic exocrine cells. The latter may be important in protection from neoplastic transformation.


Next generation dengue vaccines: A review of the preclinical development pipeline.
Dengue represents a significant and growing public health problem across the globe, with approximately half of the world's population at risk. The increasing and expanding burden of dengue has highlighted the need for new tools to prevent dengue, including development of dengue vaccines. Recently, the first dengue vaccine candidate was evaluated in Phase 3 clinical trials, and other vaccine candidates are under clinical evaluation. There are also a number of candidates in preclinical development, based on diverse technologies, with promising results in animal models and likely to move into clinical trials and could eventually be next-generation dengue vaccines. This review provides an overview of the various technological approaches to dengue vaccine development with specific focus on candidates in preclinical development and with evaluation in non-human primates.


What are the remaining limitations of TAVI?
Transcatheter aortic valve implantation (TAVI) is a recognized therapy for patients with symptomatic severe aortic stenosis (AS). TAVI is superior compared to medical therapy as for mortality in extreme-risk patients, is non-inferior or superior to surgery in high-risk patients and non-inferior to surgery in intermediate-risk patients. However, several limitations affect outcomes after TAVI. Adverse events related to this procedure, like vascular complications, need for pacemaker implantation, paravalvular regurgitation, can be factors limiting TAVI treatment in younger patients at lower risk, as well as uncertainties regarding valve durability. This review tries to figure out some of the main complications still unsolved after TAVI.


[On the usefulness of inscription registries in the health professions for local history of pharmacy: the pharmacists of the arrondissement of Dreux from 1847-1941].
The author calls attention to an unacknowledged source of information for local history of pharmacy in the 19th and 20th centuries: the inscription registries of members of the health professions authorized to practice. He takes as example the arrondissement of Dreux (Eure-et-Loir) from which he draws up a list of pharmacists from 1847, their dates and places of birth, the dates and sites of the granting of their diplomas and the towns in which they practiced.


Evaluation of older persons with mild cognitive deficits: potential utility of magnetic resonance imaging.
This study evaluated brain volumes in healthy older subjects without dementia who presented with memory complaints. The objective was to examine cortical volumes in relation to cognitive performance among patients who do not have dementia, but who do have mild cognitive deficits. Fifteen participants were evaluated (mean age = 71.8 +/- 6.2). Brain structure was measured via high-resolution magnetic resonance imaging to quantify gray and white matter volumes. Volumetric measures were assessed relative to cognitive function in separate regression models controlling for total cerebral volume. Reported here are measures of global cognitive performance using the Mattis Dementia Rating Scale (DRS) in relation to volumetric measures. Baseline MMSE scores ranged from 27 to 30 (mean = 29.3; SD = 0.9). After controlling for total cerebral volume, we observed that lower white matter volume in the temporal lobe [F(1,14) = 5.72, p = 0.03] was associated with lower performance on the Mattis Dementia Rating Scale (DRS). Structural imaging may help provide useful clinical information in the context of mild cognitive decline. Currently, the diagnosis of dementia relies on longitudinal measures of cognition. Future studies will help determine whether the addition of brain imaging may enhance diagnostic certainty as well as predict long-term outcome.


Polyamine biosynthesis enzymes in the induction and inhibition of differentiation in Friend erythroleukemia cells.
O-Ornithine decarboxylase (ODC) activity in cultures of Friend erythroleukemia cells induced to differentiate with various compounds was examined. Based on ability to stimulate ODC activity, the inducers tested could be divided into two categories. Inducers of the first class, among which were dimethyl sulfoxide and hexamethylene bisacetamide, stimulated ODC activity and were inhibited by dexamethasone and the phorbol diester, 12-O-tetradecanoylphorbol-13-acetate. Specific inhibitors of polyamine biosynthesis, such as methylglyoxal bis(guanylhydrazone), an inhibitor of S-adenosylmethionine decarboxylase, and the ornithine analogs, alpha-methylornithine and alpha-hydrazinoornithine, also inhibited the induction of Friend erythroleukemia cell differentiation. The inhibition of induced differentiation by this class of compounds could be abrogated by spermine, spermidine, or putrescine. Inducers of the second class, among which were sodium butyrate, actinomycin D, and aminonucleoside of puromycin, had little or no stimulatory effect on ODC and were inhibited only by bromodeoxyuridine. The effect of bromodeoxyuridine, which inhibits inducers of both classes, was not abrogated by polyamines.


Basic residues in azurocidin/HBP contribute to both heparin binding and antimicrobial activity.
Azurocidin/CAP37/HBP is an antimicrobial and chemotactic protein that is part of the innate defenses of human neutrophils. In addition, azurocidin is an inactive serine protease homolog with binding sites for diverse ligands including heparin and the bovine pancreatic trypsin inhibitor (BPTI). The structure of the protein reveals a highly cationic domain concentrated on one side of the molecule and responsible for its strong polarity. To investigate the role of this highly basic region, we produced three recombinant azurocidin mutant proteins that were altered in either one or both of two clusters of 4 basic residues located symmetrically on each side of a central cleft in the cationic domain. Two of the mutant proteins (Loop 3: R5Q, K6Q, R8Q, and R10Q; Loop 4: R61Q, R62Q, R63Q, and R65Q) exhibited little or no change in heparin and BPTI binding or in antimicrobial function. In contrast, the Loop 3/Loop 4 mutant (R5Q, K6Q, R8Q, R10Q, R61Q, R62Q, R63Q, and R65Q) in which all 8 basic residues were replaced showed greatly decreased ability to bind heparin and to kill Escherichia coli and Candida albicans. Thus, we report that the 8 basic residues that were altered in the Loop 3/Loop 4 mutant contribute to the ability of the wild-type azurocidin molecule to bind heparin and to kill E. coli and C. albicans. Because BPTI binding was comparable in wild-type and Loop 3/Loop 4 mutant protein, we conclude that the same 8 basic residues are not involved in the binding of BPTI to azurocidin, supporting the notion that the binding site for BPTI is distinct from the site involved in heparin binding and antimicrobial activity. Finally, we show that removal of all 4 positively charged amino acids in the 20-44 azurocidin sequence (DMC1: R23Q,H24S,H32S,R34Q), a region previously thought to contain an antimicrobial domain, does not affect the activity of the protein against E. coli, Streptococcus faecalis, and C. albicans.


Hybrid functionals with local range separation.
Range-separated (screened) hybrid functionals provide a powerful strategy for incorporating nonlocal exact (Hartree-Fock-type) exchange into density functional theory. Existing implementations of range separation use a fixed system-independent screening parameter. Here, we propose a novel method that uses a position-dependent screening function. These locally range-separated hybrids add substantial flexibility for describing diverse electronic structures and satisfy a high-density scaling constraint better than the fixed screening approximation does.


Predictors of pregnancy among young people in sub-Saharan Africa: a systematic review and narrative synthesis.
Sub-Saharan Africa has among the highest prevalence of teenage pregnancy in the world. Teenage mothers and their children are at risk to a host of medical, social and economic challenges. Adolescent pregnancy is a significant cost to the mother and newborn child, and also to their family and the wider society. Despite measures taken by some sub-Saharan nations to tackle the issue of adolescent pregnancy, the phenomenon remains a public health concern that is widespread throughout the region. Currently, there are few studies that examine the predictors of teenage pregnancy in the sub-Saharan region. The objective of the present study was to systematically review predictors of pregnancy among young people in sub-Saharan Africa. A literature search was conducted using Medline, CINAHL and EMBASE electronic databases. Following duplicate removal, abstract and full-text screening, 15 studies were ultimately included in the final review. Narrative synthesis was used to synthesise the qualitative and quantitative findings. This review followed the Preferred Reporting Items for Systematic Reviews and Meta-analyses. Twenty-seven predictors of teenage pregnancy were identified and grouped into six themes (Partner and peer-related predictors; Sexual health knowledge, attitude and behaviour-related predictors; Parenting and family-related predictors; Economic, environmental and cultural predictors; Personal predictors; and Quality of healthcare services predictors). The most obvious predictors included sexual coercion and pressure from male partners, low or incorrect use of contraceptives, and poor parenting or low parental communication and support. This review emphasises that the large prevalence of adolescent pregnancy in sub-Saharan Africa is attributable to multiple predictors that our study was able to group into six themes. Policy changes and programmes must be implemented in sub-Saharan Africa to address these determinants in order to reduce adolescent pregnancy within the region.


Triglycerides and coronary risk in women and the elderly.
There seems little doubt that triglycerides are causally related to the progress of atherogenesis. Mechanisms for this effect include adverse quantitative and qualitative changes in circulating lipoproteins. In particular, the effects of lower high-density lipoprotein levels and the production by hypertriglyceridemia of small, dense low-density lipoproteins are of great significance. The role of triglyceride-rich remnant particles in atherogenesis is likely important. These remnants, which are lipoproteins rich in both cholesterol and triglycerides, can be shown to produce cholesteryl ester-laden macrophages in vitro and are probably atherogenic in vivo. Triglyceride levels are a significant risk factor for coronary artery disease in women, more so than in men. Triglyceride levels also increase in older patients and continue to be predictors of coronary risk in both men and women older than 65 years. It is unclear whether triglyceride intervention efforts should be directed at lowering triglyceride levels (such as is accomplished with niacin or fibric acid derivatives) or lowering low-density lipoprotein levels in patients with high triglyceride levels, assuming triglyceride levels are only a passive marker of atherosclerotic risk. Until more is known about the precise role of hypertriglyceridemia in atherogenesis in women and older patients, use of triglyceride-lowering drugs should be conservative and limited to those individuals with high triglyceride levels (> 4.5 mmol/L [> 400 mg/dL]) who do not respond to diet modifications and who are at risk of coronary disease either because of a history of vascular disease or the presence of other risk factors.


In vivo effects of cortisone on the B cell line in chickens.
The effects of a single dose of cortisone acetate (5 or 10 mg/100 g body weight) on B cells were examined in young chickens. A dose-dependent increase in numbers of circulating B lymphocytes and a change in their Ig-class distribution were followed by parallel increase in splenic plasma cells and serum immunoglobulins. The higher dose of cortisone produced changes in Bmu and Bgamma cells, whereas the lower dose primarily affected Bmu cells. These steroid-induced changes were preceded by lymphocyte depletion in the cortical regions of bursal follicles, and prior bursectomy prevented steroid-induced increases in circulating B lymphocytes and tissue plasma cells. The results suggest that cortisone can induce bursal lymphocytes to migrate from the bursa and to settle subsequently in peripheral lymphoid tissues where they become mature plasma cells.


T Follicular Regulatory Cell Suppression of T Follicular Helper Cell Function Is Context-Dependent in vitro.
The production of antibody-secreting plasma cells and memory B cells requires the interaction of T follicular helper (Tfh) cells with B cells in the follicle and is modulated by T follicular regulatory (Tfr) cells. We compare the effects of Tfr cells in an in vitro model of bystander Tfh function in the absence of BCR engagement and in a model in which mimics cognate T-B interactions in which the BCR is engaged. In the absence of Tfr cells, Tfh cells from primed mice induce naive B cell differentiation into GC B cells and class switch recombination (CSR) in the presence of anti-CD3 alone or anti-CD3/IgM in a contact-dependent manner. Addition of primed Tfr cells efficiently suppressed GC B cell proliferation, differentiation and CSR in the anti-CD3 alone cultures, but only moderately suppressed BCR-stimulated B cells. When stimulated with anti-CD3 alone, IL-4 is critical for the induction of GC B cells and CSR. IL-21 plays a minimal role in GC B cell differentiation, but a greater role in switching. When the BCR is engaged, IL-4 is primarily required for switching and IL-21 only modestly affects switching. CD40L expression was critical for Tfh-mediated B cell proliferation/differentiation in the absence of B cell engagement. When the BCR was engaged, proliferation of CD40 deficient B cells was partially restored, but was susceptible to suppression by Tfr. These studies suggest that in vitro Tfr suppressor function is complex and is modulated by BCR signaling and CD40-CD40L interactions.


Agnostic-Histology Approval of New Drugs in Oncology: Are We Already There?
Over the last several years, several molecular aberrations have been unevenly described across cancers, although the distinct functional relevance in each biological context is not yet fully understood. Novel discoveries have led to the development of drugs tailored to the molecular profile of patients, thus increasing the likelihood of response among biomarker-selected patients. In this context, there has been a progressive redefinition of a precision medicine framework where evidence-based development and earlier approvals might now be driven by this molecular information. Innovative trial designs have greatly facilitated the evaluation and approval of new drugs in small cohorts of orphan cancers in which histology-dependent molecularly defined trials might be logistically difficult. However, accelerated approvals based on this agnostic-histology development model have brought new clinical, regulatory, and reimbursement challenges. In this article, we will highlight many of the biologic issues and clinical trial design challenges characterizing the development of tissue-agnostic compounds. Also, we will review some of the key factors involved in the development of pembrolizumab and larotrectinib, the first two drugs that have been approved by the U.S. Food and Drug Administration in an histology-agnostic manner. Because we anticipate that agnostic-histology approvals will continue to grow, we aim to provide insight into the current panorama of targeted drugs that are following this strategy and some premises to take into consideration. Clinicians and regulators should be prepared to overcome the associated potential hurdles, ensuring that uncertainties are dealt with properly and allowing new, promising agents to arrive faster to the market.


Retinal vascular caliber changes after intravitreal triamcinolone treatment for diabetic macular edema.
To describe the changes in retinal vascular caliber in response to a single injection of intravitreal triamcinolone (IVTA) in patients with refractory diabetic macular edema. Prospective data from a randomized clinical trial were used. The effects of IVTA versus sham injections in patients with refractory diabetic macular edema were evaluated in a randomized clinical trial involving 69 eyes of 43 patients. Of these, 28 eyes (15 IVTA and 13 sham) of 21 patients had gradable retinal photographs at the baseline and 3-month follow-up visit for analysis in the present study. Retinal vascular caliber was measured from digital fundus photographs and summarized as central retinal artery (CRAE) and vein (CRVE) equivalents in all eyes at baseline and at the 3-month follow-up visit. Over the 3 months of the study, there was a significant reduction compared to baseline in retinal arteriolar (147.8 microm vs. 140.0 microm, P = 0.047) and venular (219.5 microm vs. 198 microm, P = 0.039) caliber in eyes treated with IVTA. There was no change in retinal arteriolar (139.9 microm vs. 139.2 microm, P = 0.878) or venular (220.3 microm vs. 217.6 microm, P = 0.534) caliber in those treated with sham injections. IVTA has a significant narrowing effect on both retinal arteriolar and venular diameter in eyes with diabetic macular edema (ClinicalTrials.gov number, NCT00148330).


Persistence of antibiotic resistance and plasmid-associated genes in soil following application of sewage sludge and abundance on vegetables at harvest.
Sewage sludge recovered from wastewater treatment plants contains antibiotic residues and is rich in antibiotic resistance genes, selected for and enriched in the digestive tracts of human using antibiotics. The use of sewage sludge as a crop fertilizer constitutes a potential route of human exposure to antibiotic resistance genes through consumption of contaminated crops. Several gene targets associated with antibiotic resistance (catA1, catB3, ereA, ereB, erm(B), str(A), str(B), qnrD, sul1, and mphA), mobile genetic elements (int1, mobA, IncW repA, IncP1 groups -α, -β, -δ, -γ, -ε), and bacterial 16S rRNA (rrnS) were quantified by qPCR from soil and vegetable samples obtained from unamended and sludge-amended plots at an experimental field in London, Ontario. The qPCR data reveals an increase in abundance of gene targets in the soil and vegetables samples, indicating that there is potential for additional crop exposure to antibiotic resistance genes carried within sewage sludge following field application. It is therefore advisable to allow an appropriate delay period before harvesting of vegetables for human consumption.


Visual orientation and self-perceptions of children in an external-cue-relevant or cue-irrelevant task situation.
On the basis of visual orientation on a block design task, 180 kindergarten through fifth-grade children were assigned to 3 categories of task approaches: (a) inner directed or task oriented (TO), (b) information seeking (IS), and (c) approval seeking or cue dependent (CD). The performance and self-perceptions of the children were then examined as they worked on a concept-identification task in which an external cue was either relevant or irrelevant. The results showed that performance of the children in the 3 categories on the task was differentially affected by the manipulated cue conditions, though not consistently across grade levels. The relationship between task-approach categories and self-perceptions also differed over grade levels.


Alkaline extraction of human plasma proteins from nondenaturing micro-2-D gels for protein/polypeptide mass measurement and peptide mass fingerprinting using MALDI-TOF MS.
Previously, we have reported a high-efficiency method of protein extraction from CBB-stained polyacrylamide gels for molecular mass measurement with MALDI-TOF MS [1]. In the present work, the alkaline extraction method was applied to CBB-stained 2-DE gels on which human plasma proteins were separated in the absence of denaturant. In order to examine the performance of the method, ten spots with apparent molecular masses (MMapp) in the range of 65 to 1000 kDa were selected and the proteins were extracted from the gel pieces. The extracts were subjected to whole-mass measurement by MALDI-TOF MS, with and without DTT treatment. In addition, the extracts were subjected to in-solution trypsin digestion followed by MALDI-TOF MS and PMF analysis. Successful extraction of proteins from the ten spots, up to MMapp 1000 kDa, has been ascertained by the significant PMF assignment (MASCOT) with high sequence coverage of the respective proteins or polypeptides. When direct mass measurement of the extracted proteins was attempted, three spots in MMapp range 65-100 kDa provided mass peaks. Five spots in MMapp range 150-400 kDa did not give mass peaks of the intact proteins, but showed those of the constituent polypeptides after the DTT treatment. Extraction of proteins prior to trypsin digestion enabled the procedure of PMF analysis to be much simpler than the conventional in-gel digestion method, providing comparable protein scores and sequence coverage. The technique presented here suggests a new strategy for the characterization of proteins separated by nondenaturing 2-DE.


Müllerian duct anomalies: embryological development, classification, and MRI assessment.
Müllerian duct anomalies (MDA) occur due to abnormal development of the uterus, cervix, and vagina, many times affecting a woman's ability to conceive and carry a pregnancy to term. The spectrum of possible abnormalities are related to the development of two separate Müllerian systems, which then fuse and subsequently undergo degeneration of the fused segments. This multiphasic development explains the multiple variations within the scheme of MDA classification. The purpose of this article is to review the embryologic development of the Müllerian ducts, relate the development to the most commonly used classification system, and review the magnetic resonance imaging (MRI) assessment of Müllerian duct anomalies. A brief review of the treatment options, as they relate to the imaging diagnosis, will be provided as well.


Mechanism of Action of Monoclonal Antibodies That Block the Activity of the Lethal Toxin of Bacillus Anthracis.
Neutralization of the lethal toxin of Bacillus anthracis is an important topic of both fundamental medicine and practical health care, regarding the fight against highly dangerous infections. We have generated a neutralizing monoclonal antibody 1E10 against the lethal toxin of Bacillus anthracis and described the stages of receptor interaction between the protective antigen (PA) and the surface of eukaryotic cells, the formation of PA oligomers, assembly of the lethal toxin (LT), and its translocation by endocytosis into the eukaryotic cell, followed by the formation of a true pore and the release of LT into the cell cytosol. The antibody was shown to act selectively at the stage of interaction between Bacillus anthracis and the eukaryotic cell, and the mechanism of toxin-neutralizing activity of the 1E10 antibody was revealed. The interaction between the 1E10 monoclonal antibody and PA was found to lead to inhibition of the enzymatic activity of the lethal factor (LF), most likely due to a disruption of true pore formation by PA, which blocks the release of LF into the cytosol.


Surviving Parents' Influence on Adult Children's Depressive Symptoms Following the Death of a First Parent.
Parents and children are linked across the life course, and they share common experiences. This article focuses on the bereavement experience of adult children's loss of a first parent during adulthood and examines the downward influence of emotional closeness with a surviving parent on adult children's depressive symptoms following loss. Analyses are based on adult children who experienced the death of a first parent (N = 227), drawn from the Longitudinal Study of Generations, a study of three-and four-generation families from Southern California. Multilevel lagged dependent variable models indicate that an emotionally close relationship with a surviving parent is related with fewer post-bereavement depressive symptoms when a mother survives a father, but not vice versa. This analysis extends the theory of linked lives and highlights the mutual influence parents and children exert, as well as the complex role of gender in shaping family relationships.