Gastro-oesophageal reflux in the irritable bowel syndrome.
Symptomatic assessment and oesophageal investigations were done in 25 consecutive patients with the irritable bowel syndrome attending a gastroenterological clinic. Symptoms of gastro-oesophageal reflux, dysphagia, and a globus sensation were significantly commoner than in a control group of fracture clinic patients. Ambulatory oesophageal pH monitoring showed clearly abnormal reflux in 11 of 22 patients (50%). Nine patients had macroscopic endoscopic changes and a further 11 biopsy changes alone, of oesophagitis which was thus present in 80% overall. Lower oesophageal sphincter pressure was significantly less in irritable bowel patients than in age and sex matched controls, but upper oesophageal sphincter pressure was comparable in the two groups and disordered peristalsis was not found. Oesophageal symptoms in the irritable bowel syndrome are mainly caused by gastro-oesophageal reflux predisposed to by a subnormal lower oesophageal sphincter pressure, rather than by oesophageal spasm.


Simple and alpha,beta-unsaturated aldehydes: correct prediction of genotoxic activity through structure-activity relationship models.
Aldehydes are widespread environmental and industrial compounds, able to stimulate a range of adverse health effects (e.g., general toxicity, allergenic reactions, mutagenicity, and carcinogenicity). We have previously presented quantitative structure-activity relationships (QSARs) for the genotoxicity of simple and alpha,beta-unsaturated aliphatic aldehydes. In this study, we show that the QSAR models are able to correctly predict--based only on the knowledge of the chemical structure--the genotoxicity of other aldehydes, not considered in the development phase of the models. This adds confidence to the reliability of our QSAR models as tools for the theoretical assessment of the genotoxic hazard posed by aldehydes. The analysis of SOS Chromotest induction ability and the ease of formation of DNA adducts by the aldehydes provided further mechanistic insights.


Blood pressure changes in feeders to cerebral arteriovenous malformations during therapeutic embolization.
We recorded blood pressures in feeding arteries of cerebral arteriovenous malformations with microcatheters before, during, and after embolization in awake patients. Our embolization results corroborate reports of intraoperative measurements that show an immediate increase in blood pressure after ligation. Demonstration of abrupt hemodynamic changes gives additional credence to the theory of normal perfusion pressure breakthrough and to theories of neurologic change based on vascular steal phenomena. Furthermore, pressure changes occurred earlier than visible slowing of flow during the embolization, suggesting that this is a more sensitive guide with which to monitor the progress of the embolization than is serial angiography.


Elevated CO2 reduces copper accumulation and toxicity in the diatom Thalassiosira pseudonana.
The projected ocean acidification (OA) associated with increasing atmospheric CO2 alters seawater chemistry and hence the bio-toxicity of metal ions. However, it is still unclear how OA might affect the long-term resilience of globally important marine microalgae to anthropogenic metal stress. To explore the effect of increasing pCO2 on copper metabolism in the diatom Thalassiosira pseudonana (CCMP 1335), we employed an integrated eco-physiological, analytical chemistry, and transcriptomic approach to clarify the effect of increasing pCO2 on copper metabolism of Thalassiosira pseudonana across different temporal (short-term vs. long-term) and spatial (indoor laboratory experiments vs. outdoor mesocosms experiments) scales. We found that increasing pCO2 (1,000 and 2,000 μatm) promoted growth and photosynthesis, but decreased copper accumulation and alleviated its bio-toxicity to T. pseudonana. Transcriptomics results indicated that T. pseudonana altered the copper detoxification strategy under OA by decreasing copper uptake and enhancing copper-thiol complexation and copper efflux. Biochemical analysis further showed that the activities of the antioxidant enzymes glutathione peroxidase (GPX), catalase (CAT), and phytochelatin synthetase (PCS) were enhanced to mitigate oxidative damage of copper stress under elevated CO2. Our results provide a basis for a better understanding of the bioremediation capacity of marine primary producers, which may have profound effect on the security of seafood quality and marine ecosystem sustainability under further climate change.


Bone apposition to a titanium-zirconium alloy implant, as compared to two other titanium-containing implants.
Implants made of commercially pure titanium (cpTi) are widely and successfully used in dentistry. For certain indications, diameter-reduced Ti alloy implants with improved mechanical strength are highly desirable. The aim was to compare the osseointegration of titanium-zirconium (TiZr) and cpTi implants with a modified sandblasted and acid-etched (SLActive) surface and with a Ti6Al4V alloy that was sand-blasted and acid-washed. Cylindrical implants with two, 0.75 mm deep, circumferential grooves were placed in the maxilla of miniature pigs and allowed to heal for 1, 2, 4 and 8 weeks. Undecalcified toluidine blue-stained ground sections were produced. Surface topography, area fraction of tissue components, and bone-to-implant contact (BIC) were determined. All materials showed significantly different surface roughness parameters. The amount of new bone within the implant grooves increased over time, without significant differences between materials. However, BIC values were significantly related to the implant material and the healing period. For TiZr and cpTi implants, the BIC increased over time, reaching values of 59.38 % and 76.15 % after 2 weeks, and 74.50 % and 84.67 % after 8 weeks, respectively. In contrast, the BIC for Ti6Al4V implants peaked with 42.29 % after 2 weeks followed by a decline to 28.60 % at 8 weeks. Significantly more surface was covered by multinucleated giant cells on Ti6Al4V implants after 4 and 8 weeks. In conclusion, TiZr and cpTi implants showed faster osseointegration than Ti6Al4V implants. Both chemistry and surface topography might have influenced the results. The use of diameter-reduced TiZr implants in more challenging clinical situations warrants further documentation in long-term clinical studies.


Choroid plexus tissue perfusion and blood to CSF barrier function in rats measured with continuous arterial spin labeling.
The choroid plexus (ChP) of the cerebral ventricles is a source of cerebrospinal fluid (CSF) production and also plays a key role in immune surveillance at the level of blood-to-CSF-barrier (BCSFB). In this study, we quantify ChP blood perfusion and BCSFB mediated water exchange from arterial blood into ventricular CSF using non-invasive continuous arterial spin labelling magnetic resonance imaging (CASL-MRI). Systemic administration of anti-diuretic hormone (vasopressin) was used to validate BCSFB water flow as a metric of choroidal CSF secretory function. To further investigate the coupling between ChP blood perfusion and BCSFB water flow, we characterized the effects of two anesthetic regimens known to have large-scale differential effects on cerebral blood flow. For quantification of ChP blood perfusion a multi-compartment perfusion model was employed, and we discovered that partial volume correction improved measurement accuracy. Vasopressin significantly reduced both ChP blood perfusion and BCSFB water flow. ChP blood perfusion was significantly higher with pure isoflurane anesthesia (2-2.5%) when compared to a balanced anesthesia with dexmedetomidine and low-dose isoflurane (1.0 %), and significant correlation between ChP blood perfusion and BCSFB water flow was observed, however there was no significant difference in BCSFB water flow. In summary, here we introduce a non-invasive, robust, and spatially resolved in vivo imaging platform to quantify ChP blood perfusion as well as BCSFB water flow which can be applied to study coupling of these two key parameters in future clinical translational studies.


The effect of tobacco control policy on smoking cessation in relation to gender, age and education in Lithuania, 1994-2010.
This study aimed to evaluate the association between tobacco control policies and trends in smoking cessation according to gender, age and educational level in Lithuania in 1994-2010. The data were obtained from nine cross-sectional postal surveys conducted biennially within the framework of Finbalt Health Monitor project during 1994-2010. Each survey was based on a nationally representative random sample drawn from the National population register. The sample consisted of 3000 citizens aged 20-64 in 1994-2008 surveys and 4000 in the 2010 survey. In total, 17161 individuals participated in all surveys. The development of tobacco control policy in Lithuania was assessed using the Tobacco Control Scale (TCS). The association of the TCS scores with short-term and long-term quitting according to gender, age and education was examined using logistic regression analysis with control for secular trends. Over the last two decades, a large improvement in the development of tobacco control policy has been achieved in Lithuania. At the same time, this progress was associated with the increase in smoking cessation. A significant increase in both short-term and long-term quit ratios was found among people aged 20-44. An increase by 10 points on the TCS was associated with 17% increase in the odds of short-term quitting and with 15% increase in the odds of long-term quitting. The association between tobacco control policies and long-term quitting was stronger among younger than older people. No differential effect of tobacco control policies on smoking cessation was found in relation to gender and educational level. The improvement in Lithuanian tobacco control policies was associated with an increase in smoking cessation in long-term perspective. These policies have not only benefitted highly educated groups, but lower educated groups as well. Nonetheless, further development of comprehensive tobacco control policies is needed in order to decrease social inequalities in smoking cessation.


Evaluation of the "steal" phenomenon on the efficacy of hypoxia activated prodrug TH-302 in pancreatic cancer.
Pancreatic ductal adenocarcinomas are desmoplastic and hypoxic, both of which are associated with poor prognosis. Hypoxia-activated prodrugs (HAPs) are specifically activated in hypoxic environments to release cytotoxic or cytostatic effectors. TH-302 is a HAP that is currently being evaluated in a Phase III clinical trial in pancreatic cancer. Using animal models, we show that tumor hypoxia can be exacerbated using a vasodilator, hydralazine, improving TH-302 efficacy. Hydralazine reduces tumor blood flow through the "steal" phenomenon, in which atonal immature tumor vasculature fails to dilate in coordination with normal vasculature. We show that MIA PaCa-2 tumors exhibit a "steal" effect in response to hydralazine, resulting in decreased tumor blood flow and subsequent tumor pH reduction. The effect is not observed in SU.86.86 tumors with mature tumor vasculature, as measured by CD31 and smooth muscle actin (SMA) immunohistochemistry staining. Combination therapy of hydralazine and TH-302 resulted in a reduction in MIA PaCa-2 tumor volume growth after 18 days of treatment. These studies support a combination mechanism of action for TH-302 with a vasodilator that transiently increases tumor hypoxia.


Effect of Sofosbuvir on rats' ovaries and the possible protective role of vitamin E: biochemical and immunohistochemical study.
Hepatitis C virus (HCV) infection is a major health problem worldwide and its eradication is mandatory. Direct acting HCV polymerase inhibitors, such as Sofosbuvir (SOF), is an effective regimen. However, it has some side effects like mutagenesis, carcinogenesis, and the impairment of testicular function. It is important to evaluate the safety of SOF on the ovary, as there are no studies yet. Increasing the production of Reactive Oxygen Species (ROS), causes oxidative stress, which affects ovulation process, female reproduction, and fertility. Accumulation of SOF in the cells was demonstrated to promote ROS generation. Vitamin E (Vit E) is an antioxidant agent that has an essential role in the female reproductive system, its deficiency can cause infertility. We explored the effect of SOF treatment alone and co-treated with Vit E on ovarian ROS level and ovarian morphology experimentally using biochemical and immunohistochemical studies. Significant changes in oxidative stress markers; nitric oxide and malondialdehyde lipid peroxidation, antioxidant enzymes; catalase, super oxide dismutase, and reduced glutathione, proliferating markers; proliferation cell nuclear antigen and Ki-67 antigen and caspase 3 apoptotic marker were demonstrated. It was shown that where SOF induced oxidative stress, it also aggravated ovarian dysfunction. The essential role of Vit E as an antioxidant agent in protecting the ovarian tissue from the effect of oxidative stress markers and preserving its function was also displayed. This could be guidance to add Vit E supplements to SOF regimens to limit its injurious effect on ovarian function.


Combining Polygenic Hazard Score With Volumetric MRI and Cognitive Measures Improves Prediction of Progression From Mild Cognitive Impairment to Alzheimer's Disease.
Improved prediction of progression to Alzheimer's Disease (AD) among older individuals with mild cognitive impairment (MCI) is of high clinical and societal importance. We recently developed a polygenic hazard score (PHS) that predicted age of AD onset above and beyond APOE. Here, we used data from the Alzheimer's Disease Neuroimaging Initiative (ADNI) to further explore the potential clinical utility of PHS for predicting AD development in older adults with MCI. We examined the predictive value of PHS alone and in combination with baseline structural magnetic resonance imaging (MRI) data on performance on the Mini-Mental State Exam (MMSE). In survival analyses, PHS significantly predicted time to progression from MCI to AD over 120 months (p = 1.07e-5), and PHS was significantly more predictive than APOE alone (p = 0.015). Combining PHS with baseline brain atrophy score and/or MMSE score significantly improved prediction compared to models without PHS (three-factor model p = 4.28e-17). Prediction model accuracies, sensitivities and area under the curve were also improved by including PHS in the model, compared to only using atrophy score and MMSE. Further, using linear mixed-effect modeling, PHS improved the prediction of change in the Clinical Dementia Rating-Sum of Boxes (CDR-SB) score and MMSE over 36 months in patients with MCI at baseline, beyond both APOE and baseline levels of brain atrophy. These results illustrate the potential clinical utility of PHS for assessment of risk for AD progression among individuals with MCI both alone, or in conjunction with clinical measures of prodromal disease including measures of cognitive function and regional brain atrophy.


The prevalence, characteristics of and risk factors for eczema in Belgian schoolchildren.
Childhood eczema is common in infants, but its nature and extent during later childhood remains unclear. In this cross-sectional study we examined the prevalence and characteristics of eczema in an unbiased community population of 2,021 Belgian schoolchildren, aged 3.4 to 14.8 years with skin prick testing and parental questionnaires. Our study identified an eczema prevalence of 23.3% and a considerable allergic comorbidity, mainly in sensitized children. The reported prevalence of eczema in infancy was 18.5% and for current eczema 11.6%. The overall sensitization rate (33.2%) as well as sensitization rates for the individual allergens were significantly higher in children with "eczema ever." Sensitization to Dermatophagoides pteronyssinus (19.6%), mixed grass pollen (15.1%), and cat (9.1%) were most common. Until the age of 6 years, boys with eczema were significantly more sensitized than girls (p = 0.007). Children with both eczema in infancy and current eczema show a tendency to be more sensitized than children with eczema in infancy only or current eczema only, but significance was only noted for a few individual allergens. Analysis of factors associated with eczema revealed a predominantly atopic profile characterized by family or personal history of allergy. Breastfeeding and environmental factors seemed to assume little relevance except for a protective effect of prematurity and having a dog at birth.


Genome-wide detection of DNA double-stranded breaks induced by engineered nucleases.
Although great progress has been made in the characterization of the off-target effects of engineered nucleases, sensitive and unbiased genome-wide methods for the detection of off-target cleavage events and potential collateral damage are still lacking. Here we describe a linear amplification-mediated modification of a previously published high-throughput, genome-wide, translocation sequencing (HTGTS) method that robustly detects DNA double-stranded breaks (DSBs) generated by engineered nucleases across the human genome based on their translocation to other endogenous or ectopic DSBs. HTGTS with different Cas9:sgRNA or TALEN nucleases revealed off-target hotspot numbers for given nucleases that ranged from a few or none to dozens or more, and extended the number of known off-targets for certain previously characterized nucleases more than tenfold. We also identified translocations between bona fide nuclease targets on homologous chromosomes, an undesired collateral effect that has not been described previously. Finally, HTGTS confirmed that the Cas9D10A paired nickase approach suppresses off-target cleavage genome-wide.


Effects of temperature on intraspecific competition in ectotherms.
Understanding how temperature influences population regulation through its effects on intraspecific competition is an important question for which there is currently little theory or data. Here we develop a theoretical framework for elucidating temperature effects on competition that integrates mechanistic descriptions of life-history trait responses to temperature with population models that realistically capture the variable developmental delays that characterize ectotherm life cycles. This framework yields testable comparative predictions about how intraspecific competition affects reproduction, development, and mortality under alternative hypotheses about the temperature dependence of competition. The key finding is that ectotherm population regulation in seasonal environments depends crucially on the mechanisms by which temperature affects competition. When competition is strongest at temperatures optimal for reproduction, effects of temperature and competition act antagonistically, leading to more complex dynamics than when competition is temperature independent. When the strength of competition increases with temperature past the optimal temperature for reproduction, effects of temperature and competition act synergistically, leading to dynamics qualitatively similar to those when competition is temperature independent. Paradoxically, antagonistic effects yield a higher population floor despite greater fluctuations. These findings have important implications for predicting effects of climate warming on population regulation. Synergistic effects of temperature and competition can predispose populations to stochastic extinction by lowering minimum population sizes, while antagonistic effects can increase the potential for population outbreaks through greater fluctuations in abundance.


Functional relationships of FANCC to homologous recombination, translesion synthesis, and BLM.
Some of the restarting events of stalled replication forks lead to sister chromatid exchange (SCE) as a result of homologous recombination (HR) repair with crossing over. The rate of SCE is elevated by the loss of BLM helicase or by a defect in translesion synthesis (TLS). We found that spontaneous SCE levels were elevated approximately 2-fold in chicken DT40 cells deficient in Fanconi anemia (FA) gene FANCC. To investigate the mechanism of the elevated SCE, we deleted FANCC in cells lacking Rad51 paralog XRCC3, TLS factor RAD18, or BLM. The increased SCE in fancc cells required Xrcc3, whereas the fancc/rad18 double mutant exhibited higher SCE than either single mutant. Unexpectedly, SCE in the fancc/blm mutant was similar to that in blm cells, indicating functional linkage between FANCC and BLM. Furthermore, MMC-induced formation of GFP-BLM nuclear foci was severely compromised in both human and chicken fancc or fancd2 cells. Our cell survival data suggest that the FA proteins serve to facilitate HR, but not global TLS, during crosslink repair.


Importance of the CCR5-CCL5 axis for mucosal Trypanosoma cruzi protection and B cell activation.
Trypanosoma cruzi is an intracellular parasite and the causative agent of Chagas disease. Previous work has shown that the chemokine receptor CCR5 plays a role in systemic T. cruzi protection. We evaluated the importance of CCR5 and CCL5 for mucosal protection against natural oral and conjunctival T. cruzi challenges. T. cruzi-immune CCR5(-/-) and wild-type C57BL/6 mice were generated by repeated infectious challenges with T. cruzi. CCR5(-/-) and wild-type mice developed equivalent levels of cellular, humoral, and protective mucosal responses. However, CCR5(-/-)-immune mice produced increased levels of CCL5 in protected gastric tissues, suggesting compensatory signaling through additional receptors. Neutralization of CCL5 in CCR5(-/-)-immune mice resulted in decreased mucosal inflammatory responses, reduced T. cruzi-specific Ab-secreting cells, and significantly less mucosal T. cruzi protection, confirming an important role for CCL5 in optimal immune control of T. cruzi replication at the point of initial mucosal invasion. To investigate further the mechanism responsible for mucosal protection mediated by CCL5-CCR5 signaling, we evaluated the effects of CCL5 on B cells. CCL5 enhanced proliferation and IgM secretion in highly purified B cells triggered by suboptimal doses of LPS. In addition, neutralization of endogenous CCL5 inhibited B cell proliferation and IgM secretion during stimulation of highly purified B cells, indicating that B cell production of CCL5 has important autocrine effects. These findings demonstrate direct effects of CCL5 on B cells, with significant implications for the development of mucosal adjuvants, and further suggest that CCL5 may be important as a general B cell coactivator.


Tuberculosis. A chemotherapeutic triumph but a persistent socioeconomic problem.
There is evidence that man has suffered from tuberculosis for more than 5,000 years, and through crowded living conditions, debilitation, and malnutrition, tuberculosis became epidemic in Western civilization and was a major cause of mortality. Identification of the tubercle bacillus as the causative agent in 1882 firmly established the infectious nature of the disease and the development of sanatoriums soon followed. Before the advent of effective chemotherapeutic agents, treatment involved rest, diet, and various surgical procedures, which were of little or no benefit to the patient. The discovery of dihydrostreptomycin, aminosalicylic acid, and isoniazid in the late 1940s and early 1950s meant that tuberculosis was now entirely curable in virtually all patients. Despite these effective chemotherapeutic and preventive agents, tuberculosis has receded to socioeconomically disadvantaged urban and rural areas, where the incidence parallels that of developing countries. Conquest of the disease will require improved health care delivery to the indigent and dispossessed.


Design principles and performance of bioresorbable polymeric vascular scaffolds.
Bioresorbable polymeric vascular scaffolds may spawn a fourth revolution in percutaneous coronary intervention (PCI) and a novel treatment termed vascular restoration therapy. The principal design considerations for bioresorbable scaffolds are discussed in the context of physiological behaviour using the Bioabsorbable Vascular Solutions (BVS) ABSORB Cohort B scaffold (Abbott Vascular) as an example. The lifecycle of a bioresorbable scaffold is divided into three phases: (1) revascularisation; (2) restoration; and (3) resorption. In the revascularisation phase spanning the first three months after intervention, the bioresorbable scaffold should perform comparably to metallic drug-eluting stents (DES) in terms of deliverability, radial strength, recoil, and neointimal thickening. The ensuing restoration phase is characterised by gradual erosion of radial strength and a loss of structural continuity, where the time scale at which each occurs is related to the hydrolytic degradation rate of the polymer. Natural vasomotion in response to external stimuli is theoretically possible at the end of this phase. Finally, in the resorption phase, the passive implant is systematically resorbed and processed by the body. Limited clinical data speak to the potential of bioresorbable scaffolds as a new therapy, and future studies will prove critical to inspiring a fourth revolution in PCI.


Identification and evolutionary implications of neurotransmitter-ciliary interactions underlying the behavioral response to hypoxia in Lymnaea stagnalis embryos.
Acceleration of embryonic rotation is a common response to hypoxia among pond snails. It was first characterized in Helisoma trivolvis embryos, which have a pair of sensorimotor neurons that detect hypoxia and release serotonin onto postsynaptic ciliary cells. The objective of the present study was to determine how the hypoxia response is mediated in Lymnaea stagnalis, which differ from H. trivolvis by having both serotonergic and dopaminergic neurons, and morphologically distinct ciliated structures at comparative stages of embryonic development. Time-lapse video recordings of the rotational behavior in L. stagnalis revealed similar rotational features to those previously observed in H. trivolvis, including rotational surges and rotational responses to hypoxia. Serotonin and dopamine increased the rate of rotation with similar potency. In contrast, serotonin was more potent than dopamine in stimulating the ciliary beat frequency of isolated pedal cilia. Isolated apical plate cilia displayed an irregular pattern of ciliary beating that precluded the measurement of ciliary beat frequency. A qualitative assessment of ciliary beating revealed that both serotonin and dopamine were able to stimulate apical plate cilia. The ciliary responses to dopamine were reversible in both pedal and apical plate cilia, whereas the responses to serotonin were only reversible at concentrations below 100 μmol l(-1). Mianserin, a serotonin receptor antagonist, and SKF83566, a dopamine receptor antagonist, effectively blocked the rotational responses to serotonin and dopamine, respectively. The rotational response to hypoxia was only partially blocked by mianserin, but was fully blocked by SKF83566. These data suggest that, despite the ability of serotonin to stimulate ciliary beating in L. stagnalis embryos, the rotational response to hypoxia is primarily mediated by the transient apical catecholaminergic neurons that innervate the ciliated apical plate.


Uncertainty in fast task-irrelevant perceptual learning boosts learning of images in women but not men.
A key tenet of models of reinforcement learning is that learning is most desirable in the times of maximum uncertainty. Here we examine the role of uncertainty in the paradigm of fast task-irrelevant perceptual learning (fast-TIPL), where stimuli that are consistently presented at relevant points in times (e.g., with task targets or rewards) are better encoded than when presented at other times. We manipulated two forms of uncertainty, expected uncertainty and unexpected uncertainty (Yu & Dayan, 2005), and compared fast-TIPL under uncertainty with fast-TIPL under no uncertainty. Results indicate a larger fast-TIPL effect under uncertainty than under no uncertainty without a difference between expected and unexpected uncertainty. However, interestingly, this effect of uncertainty on fast-TIPL was found in women but not in men. In men, equivalent fast-TIPL was observed under no uncertainty and uncertainty, whereas in women, confirming previous results (Leclercq & Seitz, 2012b), no fast-TIPL was observed in the no-uncertainty condition, but fast-TIPL was observed in the uncertainty conditions. We discuss how these results imply differences in attention or neuromodulatory processes between men and women.


Performance of amplified DNA in an Illumina GoldenGate BeadArray assay.
Whole genome amplification (WGA) offers a means to enrich DNA quantities for epidemiologic studies. We used an ovarian cancer study of 1,536 single nucleotide polymorphisms (SNPs) and 2,368 samples to assess performance of multiple displacement amplification (MDA) WGA using an Illumina GoldenGate BeadArray. Initial screening revealed successful genotyping for 93.4% of WGA samples and 99.3% of genomic samples, and 93.2% of SNPs for WGA samples and 96.3% of SNPs for genomic samples. SNP failure was predicted by Illumina-provided designability rank, %GC (P < or = 0.002), and for WGA only, distance to telomere and Illumina-provided SNP score (P < or = 0.002). Distance to telomere and %GC were highly correlated; adjustment for %GC removed the association between distance to telomere and SNP failure. Although universally high, per-SNP call rates were related to designability rank, SNP score, %GC, minor allele frequency, distance to telomere (P < or = 0.01), and, for WGA only, Illumina-provided validation class (P < 0.001). We found excellent concordance generally (>99.0%) among 124 WGA:genomic replicates, 15 WGA replicates, 88 replicate aliquots of the same WGA preparation, and 25 genomic replicates. Where there was discordance, it was across WGA:genomic replicates but limited to only a few samples among other replicates suggesting the introduction of error. Designability rank and SNP score correlated with WGA:genomic concordance (P < 0.001). In summary, use of MDA WGA DNA is feasible; however, caution is warranted regarding SNP selection and analysis. We recommend that biological SNP characteristics, notably distance to telomere and GC content (<50% GC recommended), as well as Illumina-provided metrics be considered in the creation of GoldenGate assays using MDA WGA DNA.


Monkeys (macaca mulatta and cebus apella) and human adults and children (homo sapiens) compare subsets of moving stimuli based on numerosity.
Two monkey species (Macaca mulatta and Cebus apella) and human children and adults judged the numerousness of two subsets of moving stimuli on a computer screen. Two sets of colored dots that varied in number and size were intermixed in an array in which all dots moved in random directions and speeds. Participants had to indicate which dot color was more numerous within the array. All species performed at high and comparable levels, including on trials in which the subset with the larger number of items had a smaller total area of coloration. This indicated a similarity across species to use the number of items in the subsets, and not dimensions such as area or volume, to guide decision making. Discrimination performance was constrained by the ratio between the subsets, consistent with other reports of numerousness judgments of stationary stimuli. These results indicate a similarity in numerical estimation ability for moving stimuli across primate species, and this capacity may be necessary for naturally occurring experiences in which moving stimuli must be summed.


Fertility preservation in women with cervical cancer.
Fertility preservation in women with cervical cancer is a demanding but evolving issue. Some remarkable achievements have been reached, in particular the improvement of primary and secondary prevention and the broadening of the indications for conservative surgery up to FIGO 2018 stage IB2. Natural pregnancy rate and the rate of obstetrics complications following conservative approach is satisfactory even if not optimal. On the other hand, the use of classic strategies for fertility preservation such as oocytes or ovarian cortex freezing is extremely limited, being the uterus compromised by treatment in a high proportion of cases. In fact, the availability of uterine surrogacy can play a role in the counseling and the decision-making process. The recent advent of uterus transplantation is fascinating but, at present, cannot be viewed as a realistic solution.


Necroptosis: A Novel Cell Death Modality and Its Potential Relevance for Critical Care Medicine.
Cell death is intertwined with life in development, homeostasis, pathology, and aging. Until recently, apoptosis was the best known form of programmed cell death, whereas necrosis was for a long time considered accidental owing to physicochemical injury. However, identification of crucial signaling and execution molecules, which are highly regulated, revealed that necrosis encompasses several cell death modalities that can be therapeutically targeted. The best understood form of regulated necrosis is necroptosis, which is transduced by the kinase activities of receptor interacting protein kinase-1 and receptor interacting protein kinase-3, eventually leading to the activation of mixed lineage kinase domain-like and plasma membrane permeabilization. We are only beginning to appreciate the role of necroptosis in different pathological conditions, including critical illnesses. In this review, we discuss the molecular mechanisms of necroptosis and analyze the effect of inhibiting necroptosis in experimental models of critical illnesses. In view of the identification of an increasing number of cell death modalities, we also briefly discuss the simultaneous targeting of multiple cell death modalities because, depending on the cell type and cellular conditions, various types of cell death may contribute to the pathology.


A convenient metal-free reagent for the generation and capture of trifluoromethanethiol.
O-Octadecyl-S-trifluorothiolcarbonate is a cheap and storable crystalline source of trifluoromethanethiol that can be prepared in two steps on a multigram scale from trifluoroacetic anhydride and sodium O-octadecyl-dithiocarbonate (xanthate). It reacts directly with gramines or with α-bromoketones and -esters in the presence of KF and pyrrolidine to give the corresponding trifluoromethyl sulfides in generally high yield.


Crystal structure of the M-fragment of alpha-catenin: implications for modulation of cell adhesion.
The cytoskeletal protein alpha-catenin, which shares structural similarity with vinculin, is required for cadherin-mediated cell adhesion, and functions to modulate cell adhesive strength and to link the cadherins to the actin-based cytoskeleton. Here we describe the crystal structure of a region of alpha-catenin (residues 377-633) termed the M-fragment. The M-fragment is composed of a tandem repeat of two antiparallel four-helix bundles of virtually identical architectures that are related in structure to the dimerization domain of alpha-catenin and the tail region of vinculin. These results suggest that alpha-catenin is composed of repeating antiparallel helical domains. The region of alpha-catenin previously defined as an adhesion modulation domain corresponds to the C-terminal four-helix bundle of the M-fragment, and in the crystal lattice these domains exist as dimers. Evidence for dimerization of the M-fragment of alpha-catenin in solution was detected by chemical cross-linking experiments. The tendency of the adhesion modulation domain to form dimers may explain its biological activity of promoting cell-cell adhesiveness by inducing lateral dimerization of the associated cadherin molecule.


Lifestyle-Integrated Functional Exercise for People With Dementia: A Pilot Study.
The objective of this study was to explore the feasibility and preliminary effectiveness of the Lifestyle-Integrated Functional Exercise for People with Dementia (LiFE4D) on health-related physical fitness, cognitive function, physical activity, and respiratory and upper limb functions. A randomized controlled pilot study was conducted (control group: usual care; experimental group: usual care and LiFE4D). The feasibility of LiFE4D was determined considering recruitment, protocol acceptability, adherence, and safety. Measures of health-related physical fitness, cognitive function, physical activity, and respiratory and upper limb functions were assessed at the baseline and 3 months. Twelve participants (8 [66.7%] female, 82 [72.2-84] years) were included, six per group. Recruitment was challenging. LiFE4D was acceptable with excellent adherence and no major adverse events. Cardiorespiratory endurance (effect size = 1.64, 95% confidence interval [CI; 0.33, 2.95]) and balance (effect size = 1.46, 95% CI [0.19, 2.73]) improved after LiFE4D. LiFE4D seems to be feasible and safe, and it shows potential to significantly improve the health-related physical fitness of people with dementia.


Human albumin enriched St. Thomas Hospital cardioplegic solution increases reperfusion injury in isolated perfused rat hearts.
In open heart surgery it is very important to protect the heart during the ischaemic period in terms of mortality and morbidity. Many different cardioplegic solutions are in clinical use without being tested experimentally. In this study we intended to investigate the effects of albumin addition to St. Thomas Hospital cardioplegic solution on cardiac protection to ischaemia. Rat hearts were isolated and perfused in Langendorff apparatus (n = 6 for each group). After the stabilisation period, the hearts in the control group were arrested with St. Thomas Hospital cardioplegic solution for 3 min then subjected to 30 min of global ischaemia in cardioplegic solution, this is followed by reperfusion for 10 min. In albumin groups, the experimental protocol was the same but 2.25%, 4.5% or 9% human albumin was added to the cardioplegic solution. All of the hearts were compared for their pre-ischaemic and post-ischaemic contractility, heart rate, coronary flow, LDH and CK enzyme leakage, and wet/dry weight ratio values. The contraction, heart rate (P < 0.01 for both), and coronary flow (only for the 9% albumin group, P < 0.05) values in the albumin group were less than the control group during the reperfusion period. There was no difference between groups in LDH, and CK leakage, and wet/dry weight ratio. The circulation of ischaemic hearts in the albumin group were diminished, possibly due to protein precipitation. This condition negatively affected the performance of the heart. The fact that there is no difference in enzyme leakage and wet/dry weight ratio, indicates that this event is not irreversible.


Classical Cardiovascular Risk Markers in Pregnancy and Associations to Uteroplacental Acute Atherosis.
Uteroplacental acute atherosis (AA) is a pregnancy-specific arterial lesion resembling early stages of atherosclerosis. AA is frequent in preeclamptic pregnancies, which associate with increased long-term maternal risk of atherosclerotic cardiovascular disease. We hypothesized that AA in pregnant women associates with classical risk factors for cardiovascular disease, including hypertension, hyperlipidemia, glucose intolerance, elevated C-reactive protein, age, and body mass index. We included 237 women delivered by cesarean section (healthy pregnancies, n=94; preeclampsia, n=87; pregestational and gestational diabetes mellitus, n=39; diabetes mellitus with preeclampsia, n=17). They provided blood before delivery for biomarker analyses. AA was diagnosed by immunohistochemistry in uteroplacental (decidual) tissue collected after placental removal. Statistical analyses were performed with Mann-Whitney test. Levels of traditional cardiovascular markers were not associated with decidual AA within the groups of women with normotensive pregnancies, preeclampsia, diabetes mellitus, or diabetes mellitus superimposed with preeclampsia. However, the oldest patient age quartile (36-43 years old) with AA had significantly higher levels of LDL (low-density lipoprotein) and apolipoprotein B (both P<0.01) than women of the same age without AA. AA was associated with elevated median prepregnancy/early pregnancy systolic blood pressure ( P=0.01) in the total cohort, but as preeclampsia was strongly associated with this finding ( P<0.01), this was likely caused by a large proportion of preeclamptic pregnancies in the AA group (62.7%). Our findings demonstrate that dyslipidemia associated with cardiovascular risk is a feature of uteroplacental AA in older women, not of AA in pregnancy in general.


Making it stick: use of active learning strategies in continuing medical education.
Despite the known benefits of active learning (AL), the predominate educational format in higher education is the lecture. The reasons for slow adaptation of AL in medical education are not well understood. The purpose of this survey was to determine knowledge, usage, attitudes, and barriers to AL use in academic Continuing Medical Education (CME). A 20-item questionnaire was developed and sent with a link to an online questionnaire to the Society of Academic Continuing Medical Education (SACME) listserv of ~ 350 professionals representing academic medical centers, teaching hospitals, and medical specialty societies in the United States (U.S.) and Canada. Responses were collected with SurveyMonkey® from October-November, 2019. Data were analyzed using SPSS®. Responses from 146 SACME members in 91 CME units yielded a ~ 42% survey response rate. Many respondents reported their self-perceived knowledge of AL as high. Advanced training (e.g., certificate, Master of Education degree) was positively correlated with AL knowledge. AL methods were reportedly used in half of the CME activities in the majority (80%) of institutions. Higher levels of self-perceived knowledge were correlated with an increased percentage of AL-related CME activities. Commonly perceived barriers to use of AL were presenters' lack of familiarity and a need for more time-consuming preparation. More efforts are needed to increase innovation and incorporate evidence-based AL strategies in medical education, especially to foster learner engagement, critical thinking, and problem-solving ability.


Biotensegrity: a unifying theory of biological architecture with applications to osteopathic practice, education, and research--a review and analysis.
Since its inception, osteopathic medicine has sought to identify the mechanical causes of disease and to understand the body's structure-function relationship. Research conducted during the past 25 years has demonstrated that the architectural principles of tensegrity can be applied to biological organisms (termed biotensegrity) and that these principles can demonstrate the mechanical structure-function relationship at all size scales in the human body. Further, biotensegrity at the cellular level allows the cell to mechanically sense its environment and convert mechanical signals into biochemical changes. When applied to the principles of osteopathic medicine, biotensegrity provides a conceptual understanding of the hierarchical organization of the human body and explains the body's ability to adapt to change. Further, biotensegrity explains how mechanical forces applied during osteopathic manipulative treatment could lead to effects at the cellular level, providing a platform for future research on the mechanisms of action of osteopathic manipulative treatment.


Evaluation of hydrogen-bonding and enantiomeric P2-S2 hydrophobic contacts in dynamic aspects of molecular recognition by papain.
1. 2-(N'-Acetyl-D-phenylalanyl)hydroxyethyl 2'-pyridyl disulphide (compound IV) (m.p. 59 degrees C; [alpha]D20 -6.6 degrees (c 1.2 in methanol)) was synthesized. 2. The results of a study of the pH-dependence of the second-order rate constant (k) for its reaction with the catalytic-site thiol group (Cys-25) of papain (EC 126.96.36.199) together with analogous kinetic data for the reactions of related time-dependent inhibitors, notably the L-enantiomer of compound (IV) (compound III) and the L- and D-enantiomers of 2-(N'-acetylphenylalanylamino)ethyl 2'-pyridyl disulphide (compounds I and II respectively), were used to assess the contributions of the (P1)-NH ... O = C < (Asp-158) and (P2) > C = O ... H-N-(Gly-66) hydrogen bonds and enantiomeric P2-S2 hydrophobic contacts in two manifestations of dynamic molecular recognition in papain-ligand association: (a) signalling to the catalytic-site region to provide for a (His-159)-IM(+)-H-assisted transition state and (b) the dependence of P2-S2 stereoselectivity on hydrogen-bonding interactions outside the S2 subsite. The analysis involved determination of the reactivities of individual ionization states of the reactions (pH-independent rate constants, k) and associated macroscopic pKa values and difference kinetic specificity energies (delta delta GKS = -RT1n(k1/k2), where k1 is the pH-independent second-order rate constant for reaction with one inhibitor and k2 is the analogous rate constant in the same ionization state for reaction with another inhibitor so that, when the structural change provides that k2 > k1, delta delta GKS is positive. 3. The kinetic data further illuminate the nature of the interdependence of binding interactions in papain first noted by Kowlessur, Topham, Thomas, O'Driscoll, Templeton & Brocklehurst [(1989) Biochem. J. 258, 755-764] in the S2 subsite, S1-S2 intersubsite and catalytic-site regions. Of particular note is the apparent dependence of the binding of the N-Ac-D-Phe moiety on the binding of the leaving group to (His-159)-Im+H and the fact that the resulting rate enhancement is more effective when (P1)-N-H is absent than when it is present. This result revealed by kinetic analysis goes beyond the conclusion suggested by model building that it is possible to make all of the binding contacts in complexes involving the D-enantiomers [(II) and (IV)] as in those involving the L-enantiomers [(I) and (III)].(ABSTRACT TRUNCATED AT 400 WORDS)


Calciphylaxis associated with cholangiocarcinoma treated with low-molecular-weight heparin and vitamin K.
Calciphylaxis is a rare disorder of small-vessel calcification and cutaneous infarction associated with chronic renal failure. Rare cases of calciphylaxis not associated with chronic renal failure have been reported with breast cancer, hyperparathyroidism, and alcoholic cirrhosis. To our knowledge, we report the first case of calciphylaxis without chronic renal failure associated with cholangiocarcinoma and the first attempt to treat calciphylaxis with vitamin K. A 56-year-old woman presented with necrotic leg ulceration. She was treated initially with low-molecular-weight heparin, with no effect. A coagulation work-up showed vitamin K deficiency. During vitamin K therapy, the patient had fulminant progression of the calciphylaxis. She died, and an autopsy showed metastatic cholangiocarcinoma. Thrombosis and protein C deficiency have been implicated in the pathophysiology of calciphylaxis. Functional protein C deficiency may be one of several factors contributing to the development of calciphylaxis. Vitamin K therapy was ineffective in our patient and may have been detrimental.


Conserved miRNAs are candidate post-transcriptional regulators of developmental arrest in free-living and parasitic nematodes.
Animal development is complex yet surprisingly robust. Animals may develop alternative phenotypes conditional on environmental changes. Under unfavorable conditions, Caenorhabditis elegans larvae enter the dauer stage, a developmentally arrested, long-lived, and stress-resistant state. Dauer larvae of free-living nematodes and infective larvae of parasitic nematodes share many traits including a conserved endocrine signaling module (DA/DAF-12), which is essential for the formation of dauer and infective larvae. We speculated that conserved post-transcriptional regulatory mechanism might also be involved in executing the dauer and infective larvae fate. We used an unbiased sequencing strategy to characterize the microRNA (miRNA) gene complement in C. elegans, Pristionchus pacificus, and Strongyloides ratti. Our study raised the number of described miRNA genes to 257 for C. elegans, tripled the known gene set for P. pacificus to 362 miRNAs, and is the first to describe miRNAs in a Strongyloides parasite. Moreover, we found a limited core set of 24 conserved miRNA families in all three species. Interestingly, our estimated expression fold changes between dauer versus nondauer stages and infective larvae versus free-living stages reveal that despite the speed of miRNA gene set evolution in nematodes, homologous gene families with conserved "dauer-infective" expression signatures are present. These findings suggest that common post-transcriptional regulatory mechanisms are at work and that the same miRNA families play important roles in developmental arrest and long-term survival in free-living and parasitic nematodes.


The power of the web: a systematic review of studies of the influence of the internet on self-harm and suicide in young people.
There is concern that the internet is playing an increasing role in self-harm and suicide. In this study we systematically review and analyse research literature to determine whether there is evidence that the internet influences the risk of self-harm or suicide in young people. An electronic literature search was conducted using the PsycINFO, MEDLINE, EMBASE, Scopus, and CINAHL databases. Articles of interest were those that included empirical data on the internet, self-harm or suicide, and young people. The articles were initially screened based on titles and abstracts, then by review of the full publications, after which those included in the review were subjected to data extraction, thematic analysis and quality rating. Youth who self-harm or are suicidal often make use of the internet. It is most commonly used for constructive reasons such as seeking support and coping strategies, but may exert a negative influence, normalising self-harm and potentially discouraging disclosure or professional help-seeking. The internet has created channels of communication that can be misused to 'cyber-bully' peers; both cyber-bullying and general internet use have been found to correlate with increased risk of self-harm, suicidal ideation, and depression. Correlations have also been found between internet exposure and violent methods of self-harm. Internet use may exert both positive and negative effects on young people at risk of self-harm or suicide. Careful high quality research is needed to better understand how internet media may exert negative influences and should also focus on how the internet might be utilised to intervene with vulnerable young people.


NHERF1 and NHERF2 are necessary for multiple but usually separate aspects of basal and acute regulation of NHE3 activity.
Na(+)/H(+) exchanger 3 (NHE3) is expressed in the brush border (BB) of intestinal epithelial cells and accounts for the majority of neutral NaCl absorption. It has been shown that the Na(+)/H(+) exchanger regulatory factor (NHERF) family members of multi-PDZ domain-containing scaffold proteins bind to the NHE3 COOH terminus and play necessary roles in NHE3 regulation in intestinal epithelial cells. Most studies of NHE3 regulation have been in cell models in which NHERF1 and/or NHERF2 were overexpressed. We have now developed an intestinal Na(+) absorptive cell model in Caco-2/bbe cells by expressing hemagglutinin (HA)-tagged NHE3 with an adenoviral infection system. Roles of NHERF1 and NHERF2 in NHE3 regulation were determined, including inhibition by cAMP, cGMP, and Ca(2+) and stimulation by EGF, with knockdown (KD) approaches with lentivirus (Lenti)-short hairpin RNA (shRNA) and/or adenovirus (Adeno)-small interfering RNA (siRNA). Stable infection of Caco-2/bbe cells by NHERF1 or NHERF2 Lenti-shRNA significantly and specifically reduced NHERF protein expression by >80%. NHERF1 KD reduced basal NHE3 activity, while NHERF2 KD stimulated NHE3 activity. siRNA-mediated (transient) and Lenti-shRNA-mediated (stable) gene silencing of NHERF2 (but not of NHERF1) abolished cGMP- and Ca(2+)-dependent inhibition of NHE3. KD of NHERF1 or NHERF2 alone had no effect on cAMP inhibition of NHE3, but KD of both simultaneously abolished the effect of cAMP. The stimulatory effect of EGF on NHE3 was eliminated in NHERF1-KD but occurred normally in NHERF2-KD cells. These findings show that both NHERF2 and NHERF1 are involved in setting NHE3 activity. NHERF2 is necessary for cGMP-dependent protein kinase (cGK) II- and Ca(2+)-dependent inhibition of NHE3. cAMP-dependent inhibition of NHE3 activity requires either NHERF1 or NHERF2. Stimulation of NHE3 activity by EGF is NHERF1 dependent.


Necessity of molecular techniques to distinguish between Neisseria meningitidis strains isolated from patients with meningococcal disease and from their healthy contacts.
Serogroup C strains of Neisseria meningitidis were isolated from a Germany patient with severe meningococcal disease after a trip to the Czech Republic. These strains (case isolates) were characterized by classical and molecular techniques, as were other strains (carrier isolates) isolated from healthy contacts. Five of 10 carrier isolates had switched off the expression of capsular polysaccharide, as demonstrated by a serogroup-specific PCR. The two case isolates were indistinguishable by multilocus sequence typing and belonged to the ET-37 complex. The carrier isolates belonged to four different sequence types, all unrelated to that of the case strains. Pulsed-field gel electrophoresis showed that the case isolates differed from reference ET-37 complex strains from the Czech Republic and Canada as well as from all the carrier isolates. The isolate from the patient's nasopharynx was indistinguishable from the blood isolate except for a 40,000-bp chromosomal deletion that had occurred during systemic spread.


Correlation of Serum IL-17, VEGF, and Lactate Dehydrogenase (LDH) Levels with Prognosis of Gastric Cancer.
To investigate the correlation of serum interleukin-17 (IL-17), vascular endothelial growth factor (VEGF) and lactate dehydrogenase (LDH) levels with the prognosis of gastric cancer patients. From December 2018 to December 2020, 45 patients with gastric cancer treated in our hospital and 50 healthy individuals were assessed for eligibility and recruited. The eligible patients were assigned to an observation group, and the healthy subjects were assigned to a control group. Serum IL-17, LDH, and VEGF levels of the eligible participants were determined by the enzyme-linked immunosorbent assay (ELISA) and biochemical testing. The association of serum IL-7, LDH, and VEGF levels with their pathological characteristics was examined in the observation group. The correlation between serum IL-17 and VEGF was analyzed using the Pearson method, and regression models were established using COX proportional risk to explore the independent risk factors for gastric cancer. Gastric cancer was associated with higher levels of IL-17, LDH, and VEGF versus a healthy status (P < 0.05). There was no significant difference in serum IL-17, LDH, and VEGF levels between the two groups of patients with different clinical characteristics (P > 0.05). Higher tumor TNM stages resulted in significantly higher levels of IL-17, LDH, and VEGF (P < 0.05). Serum IL-17 level was positively correlated with VEGF level (P < 0.05). Cox regression multifactorial analysis showed that serum IL-17, LDH, VEGF, and tumor TNM stages could be independent high-risk influencing factors for gastric cancer (P < 0.05). Serum IL-17 was positively correlated with VEGF levels in patients with gastric cancer. Serum IL-17, LDH, and VEGF levels in gastric cancer patients are closely correlated with the TNM stage and patients' prognosis, both of which show great potential as effective indicators for evaluating the prognosis of gastric cancer.


Dietary omega-3 and polyunsaturated fatty acids modify fatty acyl composition and insulin binding in skeletal-muscle sarcolemma.
Feeding animals with diets high in saturated fat induces insulin resistance, and replacing saturated fat isocalorically with poly-unsaturated fat, especially long-chain omega-3 fatty acids, will prevent the development of insulin resistance in skeletal-muscle tissue. To investigate the mechanism, rats were fed on high-fat (20%, w/w) semipurified diets for 6 weeks. Diets containing ratios of polyunsaturated/saturated (P/S) fatty acid of 0.25 (low-P/S diet) and 1.0 (high-P/S diet) were used to study the effect of the level of saturated fat. To study the effects of omega-3 fatty acids, diets with a low-P/S ratio containing either 0 (low-omega-3 diet) or 3.3% (high-omega-3 diet) long-chain omega-3 fatty acids from fish oil were fed. Plasma membrane from skeletal muscle was purified. The content of fatty acids in sarcolemmal phospholipid was significantly related to the dietary composition. Insulin binding to intact sarcolemmal vesicles prepared from rats fed on diets high in omega-3 fatty acids increased 14-fold compared with animals fed on the low-omega-3 diet (P < 0.0001). Feeding rats on a diet with a high P/S ratio increased sarcolemmal insulin binding by 2.3-fold (P < 0.05). Increased insulin binding was due to increased receptor number at the low-affinity high-capacity binding site. Dietary effects on insulin binding were eliminated when studies were carried out on detergent-solubilized membranes, indicating the importance of the phospholipid fatty acyl composition for insulin binding. The results suggest that dietary omega-3 and polyunsaturated fatty acids increase insulin binding to sarcolemma by changing the fatty acyl composition of phospholipid surrounding the insulin receptor, and this might be the mechanism by which dietary fatty acids modify insulin action.


Beam-target double-spin asymmetry A{LT} in charged pion production from deep inelastic scattering on a transversely polarized {3}He target at 1.4<Q{2}<2.7 GeV{2}.
We report the first measurement of the double-spin asymmetry A{LT} for charged pion electroproduction in semi-inclusive deep-inelastic electron scattering on a transversely polarized {3}He target. The kinematics focused on the valence quark region, 0.16<x<0.35 with 1.4<Q{2}<2.7 GeV{2}. The corresponding neutron A{LT} asymmetries were extracted from the measured {3}He asymmetries and proton over {3}He cross section ratios using the effective polarization approximation. These new data probe the transverse momentum dependent parton distribution function g{1T}{q} and therefore provide access to quark spin-orbit correlations. Our results indicate a positive azimuthal asymmetry for π{-} production on {3}He and the neutron, while our π{+} asymmetries are consistent with zero.


Safety and practicality of high dose inhaled nitric oxide in emergency department COVID-19 patients.
Inhaled nitric oxide (iNO) is a selective pulmonary vasodilator and mild bronchodilator that has been shown to improve systemic oxygenation, but has rarely been administered in the Emergency Department (ED). In addition to its favorable pulmonary vascular effects, in-vitro studies report that NO donors can inhibit replication of viruses, including SARS Coronavirus 2 (SARS-CoV-2). This study evaluated the administration of high-dose iNO by mask in spontaneously breathing emergency department (ED) patients with respiratory symptoms attributed to Coronavirus disease 2019 (COVID-19). We designed a randomized clinical trial to determine whether 30 min of high dose iNO (250 ppm) could be safely and practically administered by emergency physicians in the ED to spontaneously-breathing patients with respiratory symptoms attributed to COVID-19. Our secondary goal was to learn if iNO could prevent the progression of mild COVID-19 to a more severe state. We enrolled 47 ED patients with acute respiratory symptoms most likely due to COVID-19: 25 of 47 (53%) were randomized to the iNO treatment group; 22 of 47 (46%) to the control group (supportive care only). All patients tolerated the administration of high-dose iNO in the ED without significant complications or symptoms. Five patients receiving iNO (16%) experienced asymptomatic methemoglobinemia (MetHb) > 5%. Thirty-four of 47 (72%) subjects tested positive for SARS-CoV-2: 19 of 34 were randomized to the iNO treatment group and 15 of 34 subjects to the control group. Seven of 19 (38%) iNO patients returned to the ED, while 4 of 15 (27%) control patients did. One patient in each study arm was hospitalized: 5% in iNO treatment and 7% in controls. One patient was intubated in the iNO group. No patients in either group died. The differences between these groups were not significant. A single dose of iNO at 250 ppm was practical and not associated with any significant adverse effects when administered in the ED by emergency physicians. Local disease control led to early study closure and prevented complete testing of COVID-19 safety and treatment outcomes measures.


Inhibition of dihydrofolate reductase: structure-activity correlations of quinazolines based upon molecular shape analysis.
A quantitative structure-activity relationship (QSAR) investigations of a set of 35 substituent-diverse quinazolines spanning an activity range of 4.16 log (1/I50) units was carried out using molecular shape analysis (MSA). I50 is the molar concentration necessary for 50% inhibiton of rat liver dihydrofolate reductase (DHFR). A correlation equation, analogous in descriptor form to those developed for two sets of 2,4-diaminotriazines which are also DHFR inhibitors, was constructed. The correlation coefficient, r, is 0.965, and the standard deviation, s, is 0.360. A second correlation equation was developed to explain the activities of the quinazolines on the basis of their shape similarity to a 2,4-diaminotriazine in its postulated active conformation as determined in a previous study. This correlation equation is again identical in descriptor form with those previously constructed for the triazines and quinazolines; r = 0.945 and s = 0.451 for this correlation equation. The ability to quantitatively explain activity in a congeneric set of compounds using a structurally diverse reference compound indicates the potential to design new lead compounds using MSA.


Prenatal immunization against experimental enteric colibacillosis in piglets.
To establish a model for the study of prenatal immunization against enteric colibacillosis a proportion of the litters of seven sows were immunized in utero 18 to 22 days before term by intra-amniotic and intramuscular injection of Escherichia coli antigen and the litters challenged at birth with either homologous or heterologous strains. Protection against homologous challenge was demonstrated in some but not all vaccinated piglets. The study was severely compromised by the occurrence of intrauterine death in a significant proportion of vaccinated piglets.


Internally displaced persons.
There were estimated to be over 20 million internally displaced persons (IDPs) at the end of 1999, a number that surpasses global estimates of refugees. Displacement exposes IDPs to new hazards and accrued vulnerability. These dynamics result in greater risk for the development of illness and death. Often, access of IDPs to health care and humanitarian assistance is excluded deliberately by conflicting parties. Furthermore, the arrival of IDPs into another community or region strains local health systems, and the host population ends up sharing the sufferings of the internally displaced. Health outcomes are dismaying. From a health perspective, the best option is to avoid human displacement. WHO contributes to the prevention of displacement by working for sustainable development. Placing health high on the political agenda helps maintain stability, and thereby, reduce the likelihood for displacement. Primary responsibility for assisting IDPs, irrespective of the cause, rests with the national government. However, where the government is unwilling or unable to provide the necessary aid, the international humanitarian community must step in, with WHO playing a major role in the health sector. There is consensus among the partners of the World Health Organization (WHO) that, in emergencies, the WHO must: 1) take the lead in rapid health assessment, epidemiological and nutritional surveillance, epidemic preparedness, essential drugs management, control of communicable diseases, and physical and psychosocial rehabilitation; and 2) provide guidelines and advice on nutritional requirements and rehabilitation, immunisation, medical relief items, and reproductive health. If the vital health needs of IDPs--security, food, water, shelter, sanitation and household items--are not satisfied, the provision of health services alone cannot save lives. Community participation is essential, and community participation implies bolstering the assets and capacities of the beneficiaries.


Genetic and environmental influences on focal brain density in bipolar disorder.
Structural neuroimaging studies suggest the presence of subtle abnormalities in the brains of patients with bipolar disorder. The influence of genetic and/or environmental factors on these brain abnormalities is unknown. To investigate the contribution of genetic and environmental factors on grey and white matter brain densities in bipolar disorder, monozygotic and dizygotic twins concordant and discordant for bipolar disorder were scanned using 1.5 Tesla magnetic resonance imaging and compared with healthy twin pairs. A total of 232 subjects: 49 affected twin pairs (8 monozygotic concordant, 15 monozygotic discordant, 4 dizygotic concordant, 22 dizygotic discordant) and 67 healthy twin pairs (39 monozygotic and 28 dizygotic) were included. After correcting for the effect of lithium, the liability for bipolar disorder was associated with decreased grey matter density in widespread areas of the brain, but most prominent in frontal and limbic regions, and with decreased white matter density in (frontal parts of) the superior longitudinal fasciculi. The genetic risk to develop bipolar disorder was related to decreased grey matter density in the right medial frontal gyrus, precentral gyrus and insula and with decreased white matter density in the superior longitudinal fasciculi bilaterally. In conclusion, pathology in the frontal lobe, especially in parts of the superior longitudinal fasciculus, may be central to the genetic risk to develop bipolar disorder, while widespread grey matter abnormalities appear related to the illness itself.


[Snow-white in danger. Women, socialization and health. Some messages, by means of allegory].
This is the modern-day allegory of Snow White, the most beautiful woman in the kingdom. Because of her beauty she is banished from the castle. The seven dwarfs who befriend her welcome her into their cottage. Little by little, they begin to impose their demands on her. They try to coerce her into submitting to life's little demands in order to attain beauty, health and happiness. Diet categorizes the foods filled with nasty calories that Snow White must learn to do without. Fitness outlines her spare time exercise regimen to keep her slim and trim. Mr. Clean provides her with specific household cleaning tips. Job tips advise her how to get a job outside the home to maintain her financial autonomy. Pills provide her with a way to assure a smooth transition between work and the household chores. Oh! And we almost forgot Baby. Who will care for Baby? Snow White of course--since she is the only woman in the house. Finally, to assure perfect co-ordination of all these tasks, Excel sends her off to take a time management course. Snow white responds enthusiastically to all the expectations. As anticipated, she loses weight. She then becomes exhausted and collapses and is transformed into Burnt-Out Beauty. She remains in a deep sleep for 100 days and nights. The dwarfs place her in a glass box in a forest clearing where she is discovered by Prince Conscience. He opens the glass box, kisses her lightly on the forehead, and immediately disappears. Snow White awakens and remembers the events preceding her deep sleep.(ABSTRACT TRUNCATED AT 250 WORDS)


Drosophila gastrulation: analysis of cell shape changes in living embryos by three-dimensional fluorescence microscopy.
The first event of Drosophila gastrulation is the formation of the ventral furrow. This process, which leads to the invagination of the mesoderm, is a classical example of epithelial folding. To understand better the cellular changes and dynamics of furrow formation, we examined living Drosophila embryos using three-dimensional time-lapse microscopy. By injecting fluorescent markers that visualize cell outlines and nuclei, we monitored changes in cell shapes and nuclear positions. We find that the ventral furrow invaginates in two phases. During the first 'preparatory' phase, many prospective furrow cells in apparently random positions gradually begin to change shape, but the curvature of the epithelium hardly changes. In the second phase, when a critical number of cells have begun to change shape, the furrow suddenly invaginates. Our results suggest that furrow formation does not result from an ordered wave of cell shape changes, contrary to a model for epithelial invagination in which a wave of apical contractions causes invagination. Instead, it appears that cells change their shape independently, in a stochastic manner, and the sum of these individual changes alters the curvature of the whole epithelium.


Medical Malpractice Claims Following Incidental Durotomy Due to Spinal Surgery.
Retrospective cohort study. Analyze medical malpractice verdicts and settlements associated with incidental durotomy. Incidental durotomy is a common complication of spine surgery. Although most intraoperative dural tears are repaired without sequelae, persistent Cerebrospinal Fluid leak, infection, or neurological injury can yield adverse outcomes. The medicolegal implications of incidental durotomy are poorly understood. Three separate, large legal databases were queried for cases involving incidental durotomy. Case, plaintiff, procedure, and outcome characteristics were analyzed. In total, 48 dural tear-related medical malpractice cases were analyzed. Most cases (56.3%) resulted in a ruling in favor of the defendant physician. Most cases alleged neurological deficits (86.7%). A large majority of cases without neurological sequelae had an outcome in favor of the defendant (83.3%). For cases involving a payment, the average amount was $2,757,298 in 2016 adjusted dollars. Additional surgery was required in 56.3% of cases, a delay in diagnosis/treatment of durotomy was present in 43.8%, and alleged improper durotomy repair was present in 22.9%. A favorable outcome for the plaintiff was more likely in cases with versus without alleged delay in diagnosis/treatment (61.9% vs. 29.6%, P = 0.025) and improper durotomy repair technique (72.7% vs. 35.1%, P = 0.040). Repeat surgery was not associated with favorable outcome for the plaintiff (42.8% cases with reoperation vs. 38.1% without, P = 0.486). This analysis of durotomy-associated closed malpractice claims after spine surgery is the largest yet conducted. Durotomy cannot always be considered an entirely benign event, and these findings have several direct implications for clinicians: late-presenting or dehiscent durotomy may be associated with adverse outcomes and subsequent risk of litigation, timely reoperation in the event of durotomy-related complications may not increase surgeon liability, and spine surgeons should be prepared to defend their choice of durotomy repair technique, should dehiscence occur. 3.


Active smoking causes oxidative stress and decreases blood melatonin levels.
Oxidative effects via free radical generation in smokers have been widely investigated. They cause lipid peroxidation, oxidation of proteins and damage to mainly lung and other tissues. In humans, antioxidative capacity of serum is related to antioxidant enzymes superoxide dismutase (SOD), glutathione peroxidase (GSH-Px) and melatonin. The effect of cigarette smoking on plasma levels of melatonin and antioxidant enzymes has not been established together yet. Also, it may not be clear if melatonin levels are affected by smoking and melatonin has a protective effect on cigarette smoking-induced free radical damage. The aim of this study is to investigate the relationship between smoking and antioxidant capacity including melatonin, a powerful endogenous antioxidant, and antioxidant enzymes in teenage girls who are active smokers. Additionally, malondialdehyde (MDA) levels were determined in those who have smoked at least one packet a day for three or more years. MDA levels have been used as a convenient index of the lipid peroxidation-related oxidative damage of tissues. Twenty-one young female active smokers who study at the School of Nursing and 21 nonsmoking students (as controls) at the same school were included in the study. The activities of two principal antioxidant enzymes SOD, GSH-Px and plasma levels of MDA were significantly increased but melatonin content of the blood was significantly decreased as compared to nonsmokers. In spite of an increase in antioxidant enzyme activities, MDA levels were slightly increased in smokers. This indicates that antioxidant self-defence mechanisms may not sufficiently protect the respiratory system from smoke-mediated oxidative injury. This result may be related to low melatonin levels in teenage female smokers. It seems that melatonin can reduce free radical damage to the respiratory system induced by cigarette smoke. Further experimental investigations with exogenous melatonin treatments will be needed.


Antimicrobial effect and physical properties of an injectable "active oxygen" gel for the treatment of periodontitis.
To evaluate an injectable gel, recently proposed for the controlled release of "active oxygen" in periodontal pockets, compared to an antibiotic or an antiseptic gel, respectively. The antimicrobial activity, injectability, texture properties, swelling and water uptake of the gels were studied. The "active oxygen" gel showed a bactericidal effect comparable to the two commercially available drug products (containing minocycline or chlorhexidine) on anaerobic periodontal pathogens and did not seem to affect aerobic strains. The gel was easy to inject and stable in an aqueous medium for several days. Texture analysis revealed potential gel fragility. The investigated gel for local delivery of oxygen can help to selectively eradicate anaerobic bacteria associated with periodontitis and promote the recovery of a healthy-compatible oral flora.


Preclinical evaluation of 18F-JNJ41510417 as a radioligand for PET imaging of phosphodiesterase-10A in the brain.
Phosphodiesterases are enzymes that inactivate the intracellular second messengers 3',5'-cyclic adenosine-monophosphate and/or cyclic guanosine-monophosphate. Of all 11 known phosphodiesterase families, phosphodiesterase-10A (PDE10A) has the most restricted distribution, with high expression in the striatum. PDE10A inhibitors are pursued as drugs for treatment of neuropsychiatric disorders. We have synthesized and evaluated (18)F-JNJ41510417 as a selective and high-affinity radioligand for in vivo brain imaging of PDE10A using PET. The biodistribution of (18)F-JNJ41510417 was evaluated in rats. Rat plasma and perfused brain homogenates were analyzed by high-performance liquid chromatography to quantify radiometabolites. Dynamic small-animal PET was performed in rats and in wild-type and PDE10A knock-out mice and compared with ex vivo autoradiography. Blocking and displacement experiments were performed using the nonradioactive analog and other selective PDE10A inhibitors. Tissue distribution studies showed predominant hepatobiliary excretion, sufficient brain uptake (0.56 ± 0.00 percentage injected dose at 2 min after tracer injection), and continuous accumulation of the tracer in the striatum over time; rapid washout of nonspecific binding from other brain regions was observed. Polar radiometabolites were detected in plasma and brain tissue. Dynamic small-animal PET showed continuous tracer accumulation in the striatum, with rapid decline in the cortex and cerebellum. Pretreatment and chase experiments with PDE10A inhibitors showed that the tracer binding to PDE10A was specific and reversible. Imaging in PDE10A knock-out and wild-type mice further confirmed that binding in the striatum was specific for PDE10A. Experiments in rats and PDE10A knock-out mice indicate that (18)F-JNJ41510417 binds specifically and reversibly to PDE10A in the striatum, suggesting that this new fluorinated quinoline derivative is a promising candidate for in vivo imaging of PDE10A using PET.


The shock of the old: hydrodynamics for the masses.
Hydrodynamic techniques such as analytical ultracentrifugation can provide key information about subunit stoichiometry and interaction strengths of protein-nucleic acid interactions. Analysis is complicated by (i) the need for low concentrations in order to observe both free and bound species and (ii) thermodynamic non-ideality. With the introduction of fluorescence optics, we are able to obtain data at lower concentrations, and improved understanding of the statistical thermodynamics of macromolecular solutions has allowed non-ideality to be accurately assigned. With these developments, it is possible now to assay protein-nucleic acid interactions at concentrations typically used in molecular biology assays.


Equilibrium sampling of persistent and bioaccumulative compounds in soil and sediment: comparison of two approaches to determine equilibrium partitioning concentrations in lipids.
The equilibrium sampling in silicone is increasingly applied to measure freely dissolved concentrations and chemical activities within bioaccumulation research of hydrophobic organic chemicals. Two equilibrium methods were applied to PCB-contaminated soil and sediment, and directly calibrated with respect to equilibrium partitioning concentrations in lipids (C(lipid,partitioning)): (i) Solid phase microextraction in the headspace above the sample (HS-SPME) required optimization for its application to PCBs, and it was calibrated above external partitioning standards in olive oil. (ii) Equilibrium sampling with internally coated glass jars with varying thicknesses of silicone (PDMS) resulted in proportionality between coating and analyte mass, which confirmed several validity criteria. C(lipid,partitioning) was here determined as product of PDMS concentration and PDMS to lipid partition ratio. The results of the two methods were in good agreement and thus validated each other. Finally, the coated glass jar method was applied to field sediment containing invertebrates, which lead to C(lipid,partitioning) that were about two times higher than measured lipid-normalized concentrations in the organisms. Temperature differences and animal lipid structure were discussed as possible reasons for this discrepancy. Both methods combine high analytical performance, reduced equilibration times and new calibration possibilities, which makes them suited for bioaccumulation research and environmental monitoring.


The influence of income on medical school admissions in Canada: a retrospective cohort study.
The socioeconomic status of applicants to Canadian medical schools has been understudied in the past two decades. Institutional efforts have been made to address the lack of socioeconomic diversity across Canada during this time. We investigated the income characteristics of medical school applicants, as well as the relationship between applicant income and offer of admission, to characterize the current state of socioeconomic diversity in medical admissions. We conducted a retrospective cohort study on 26,120 applicants at one Ontario medical school from 2013 to 2018. Characteristics of applicants who were offered admission were compared to the general population and applicants not offered admission. Regression analyses were used to investigate the association between median total neighborhood income and successful admission. The median total neighborhood income for medical school applicants was $98,816, which was approximately $28,480 higher than the Canadian general population. Those not admitted to the medical school had a median total neighborhood income of $98,304 compared to $105,984 for those offered admission (p < 0.001). This trend was seen in every province and territory in Canada. Median total neighborhood income was a predictor of an offer of admission; applicants in the >75th percentile income group had 54% increased odds of being offered admission when compared to applicants in the <25th percentile in our unadjusted model. Income was not significant in our adjusted models but showed that the income medians drastically shifted between pre-interview and post-interview periods, from $98,816 to $104,960 (p < 0.001). Medical school applicants are from higher economic strata compared to the general population. Despite already representing a high economic stratum, a higher median total neighborhood income relative to other applicants was associated with an offer of admission.


MicroRNA-770-5p is involved in the development of diabetic nephropathy through regulating podocyte apoptosis by targeting TP53 regulated inhibitor of apoptosis 1.
Diabetic nephropathy (DN) is a major diabetic micro-vascular complication, and podocyte apoptosis induced by high glucose (HG) is a typical early feature of DN. Studies have shown that microRNAs (miRNAs) play a crucial role in the pathogenesis of DN. The purpose of the current study was to explore the role and molecular mechanism of miR-770-5p in podocyte apoptosis in DN. In vitro podocyte model of DN was conducted by treatment conditionally immortalized mouse podocytes with HG (30 mM D-glucose). The level of miR-770-5p in podocytes was detected using quantitative real-time PCR (qRT-PCR), and protein levels were measured using Western blot assay in our current study. The relationship between miR-770-5p and TP53 regulated inhibitor of apoptosis 1 (TRIAP1) was revealed by TargetScan and dual luciferase reporter assay. Cell proliferation ability and cell apoptosis were determined by using cell counting kit-8 (CCK-8) assay and flow cytometer (FCM), respectively. We found that miR-770-5p was significantly upregulated in podocytes under HG condition. TRIAP1 was a target gene of miR-770-5p and it was down-regulated in podocytes by HG treatment. Further analysis indicated that HG induced cell proliferation ability reduction, cell apoptosis enhancement and apoptotic peptidase activating factor 1(APAF1)/Caspase9 pathway exaltation in podocytes were prevented by miR-770-5p down-regulation. More importantly, the results showed that all the effects of miR-770-5p inhibitor on HG induced podocytes were eliminated by TRIAP1 silencing. S.-Z. Zhang, X.-J. Qiu, S.-S. Dong, L.-N. Zhou, Y. Zhu, M.-D. Wang, L.-W. Jin We showed that miR-770-5p was upregulated in the in vitro model of DN, and it might promote the development of DN through regulating podocyte apoptosis by targeting TRIAP1.


Involvement of beta 2-adrenoceptor-mediated mechanisms in the cardiovascular responses to alpha 1- and alpha 2-adrenoceptor antagonism in conscious, unrestrained, Long Evans and Brattleboro rats.
1. Intra-arterial blood pressures and heart rates were recorded in conscious, unrestrained, Long Evans and Brattleboro rats receiving sequential, continuous administrations of selective alpha 1- (prazosin) and alpha 2- (idazoxan) adrenoceptor antagonists. The same protocols were also run in the presence of ICI 118551 (a selective antagonist of beta 2-adrenoceptors). 2. Prazosin and idazoxan caused large, but transient, hypotensions in Long Evans and Brattleboro rats. In the continued presence of both drugs there were marked, intermittent, depressor episodes and tachycardias in both strains of rat. 3. In the presence of low or high doses of ICI 118551 the hypotensive responses to prazosin and idazoxan were markedly reduced in both strains of rat and blood pressures showed little variability, although intermittent tachycardias still occurred. 4. In adrenal-demedullated Long Evans rats, the hypotensive responses to prazosin and idazoxan were attenuated and in the presence of both drugs, blood pressure was relatively steady, although intermittent tachycardias still occurred. 5. In the presence of prazosin and idazoxan, when a depressor episode was not occurring, administration of captopril caused hypotension in Long Evans and Brattleboro rats. In the latter, the reduction in blood pressure was sustained, whereas there was a recovery in blood pressure in Long Evans rats. This recovery was punctuated by depressor episodes, and was abolished by a V1-receptor antagonist (d(CH2)5DAVP). 6. Long Evans rats given two primed doses of the non-selective alpha-adrenoceptor antagonist, phentolamine, exhibited variation in blood pressure similar to that seen in the presence of prazosin and idazoxan. As in the latter case, blood pressure variability was inhibited by the beta 2-adrenoceptor antagonist, ICI 118551. 7. Administration of idazoxan into a lateral ventricle in Long Evans rats receiving phenoxybenzamine intravenously did not cause blood pressure instability. However, intravenous administration of idazoxan in the same animals produced intermittent depressor episodes and tachycardias similar to those seen in the presence of prazosin and idazoxan. 8. The simplest explanation of the results is that beta 2-adrenoceptor-mediated depressor mechanisms contribute to the hypotensive responses to alpha 1- and alpha 2-adrenoceptor antagonism. Furthermore, in the presence of adequate peripheral alpha 1- and alpha 2-adrenoceptor antagonism, blood pressure may be maintained by the renin-angiotensin system and vasopressin (although it is only when the former system has been antagonized that a clear-cut pressor action of vasopressin is apparent). Under these conditions, blood pressure maintenance is interrupted by intermittent depressor episodes that are largely due to adrenal medullary activation.


Free flap blood flow evaluated using two-dimensional laser speckle flowgraphy.
Objective. We investigated the efficiency of laser speckle flowgraphy for evaluating blood flow in free flaps used for plastic surgery. Methods. We measured blood flow using a visual laser meter capable of providing two-dimensional color graphic representations of flow distribution for a given area using a dynamic laser speckle effect. Using laser speckle flowgraphy, we examined the blood flow of 20 free flaps applied following the excision of head and neck tumors. Results. After anastomosis of the feeding and draining blood vessels and sewing the flap, musculocutaneous (MC) flaps showed significantly lower blood flow than jejunal or omental flaps (P < .05). The ratio of blood flow decrease from the edge to the center was significantly greater in MC flaps than in jejunal or omental flaps (P < .001). Conclusion. Laser speckle flowgraphy is useful for the perioperative measurement of blood flow in free flaps used in plastic surgery. This method is a highly useful, practical, and reliable tool for assessing cutaneous blood flow and is expected to be applicable to several clinical fields.


Genes of ACYL CARRIER PROTEIN Family Show Different Expression Profiles and Overexpression of ACYL CARRIER PROTEIN 5 Modulates Fatty Acid Composition and Enhances Salt Stress Tolerance in Arabidopsis.
Acyl carrier proteins (ACPs) are a group of small acidic proteins functioning as important cofactors in the de novo synthesis of fatty acids. In Arabidopsis, ACPs are encoded by a small gene family comprising five plastid members, AtACP1 to AtACP5, and three mitochondrial members. The biological functions and the transcriptional responses to abiotic stresses of most AtACPs have yet to be elucidated. The present study extends previous findings and provides new knowledge on the function of ACPs by examining the responses of AtACP-encoding genes to several abiotic stresses and, in particular, the role of AtACP5 in the adaptation to salt stress. Phylogenetic analysis showed that AtACP1, AtACP2, AtACP3, and AtACP5 can be classified into one group and separated from a group comprising AtACP4 and ACP homologs from related species. Quantitative RT-PCR analysis revealed that the expression of AtACP1, AtACP2, and AtACP3 was induced by drought. Both iron deficiency and nitrogen starvation resulted in down-regulation of AtACP4. The most pronounced response was observed for AtACP5, the expression of which was dramatically decreased by salt stress. Knock-out of AtACP5 showed increased sensitivity to NaCl stress, whereas transgenic lines overexpressing AtACP5 displayed increased salt tolerance relative to the wild-type. Overexpression of AtACP5 further led to an altered composition of fatty acids, mainly a decrease of oleic acid (C18:1) and an increase of palmitic acid (C16:0), and to a lower Na+/K+ ratio when compared to the salt stressed wild-type. The comprehensive transcriptional information on the small plastid AtACP gene family in response to various abiotic stresses and the further investigation of the AtACP5 indicate that AtACP5 might be critical for salt tolerance through alterations of the composition of fatty acids and, subsequently, the Na+/K+ ratio.


Distinguising features of left anterior fascicular block and inferior myocardial infarction as presented by body surface potential mapping.
Total body surface maps obtained from 19 patients with previous inferior myocardial infarction (IMI) were compared with maps obtained from 19 patients with left anterior fascicular block (LAFB) and six more patients in whom electrocardiographic changes were indistinguishable between IMI and LAFB. Three distinguishing features were detected: 1) abnormal high anterior positivity developed both in IMI and LAFB, but its onset was earlier in LAFB; 2) a broad rim of abnormal right lower negativity was seen in both groups, but in IMI it was within the first 40 msec, whereas in LAFB it was found in the middle and later parts of depolarization; 3) abnormal left lower negativity was seen in all the patients with LAFB, but was absent in IMI. Thus, despite similarities in the abnormalities detected, we found definite temporal and topographical differences that should aid in differentiating between IMI and LAFB in ambiguous cases.


Effect of hepatic dysfunction on oral cyclosporin pharmacokinetics in marrow transplant patients.
The effect of hepatic dysfunction, defined as abnormal serum bilirubin level, on oral cyclosporin (CSP) pharmacokinetics was examined in 28 marrow transplant patients who received CSP for prophylaxis of graft-v-host disease. Serum CSP concentrations were measured by radioimmunoassay. Forty-one concentration-time courses were studied, divided among patients with no (less than 1.2 mg/dL), mild (1.2 to 2.0 mg/dL), and moderate (2.0 to 5.0 mg/dL) hepatic dysfunction. CSP elimination, as determined by elimination rate constant and clearance, was delayed in patients with moderate hepatic dysfunction compared to those with no hepatic dysfunction (P less than .05). The volume of distribution, lag time for absorption, maximum serum concentration, and time at which the maximum concentration was achieved was not affected by hepatic function. These data indicate that patients with moderate hepatic dysfunction have delayed CSP or CSP metabolite elimination and may be at higher risk for developing CSP-related toxicity.


Peripheral blood dendritic cells in persons with AIDS and AIDS related complex: loss of high intensity class II antigen expression and function.
The antigen-specific immune response in HIV sero-positive individuals is depressed or absent. This may be due in part to abnormal co-operation between T lymphocytes and antigen presenting cells (APC). We have isolated from the peripheral blood of healthy heterosexuals and patients with persistent generalized lymphadenopathy (PGL) and AIDS, cells of low density (LDC) with dendritic morphology. These cells are known to be potent APC. The expression of two cell surface antigens on these cells, namely 63D3 (a monocyte related antigen) and Class II antigens was examined. LDC from controls and patients with benign, non-progressive PGL (type A) were found to show biphasic expression of Class II antigens. By contrast, the high intensity Class II expression seen on a small proportion of 63D3 negative cells from controls and patients with PGL type A was absent in patients with PGL type B (showing subtle signs of progressive immunodeficiency) and AIDS. The loss of this population of dendritic cells was reflected in the absence of stimulator activity in autologous and heterologous mixed lymphocyte reactions. Thus, it is possible that the loss of these dendritic cells may contribute to the profound immunological abnormalities seen in AIDS.


Glycerol Solvates DPPC Headgroups and Localizes in the Interfacial Regions of Model Pulmonary Interfaces Altering Bilayer Structure.
The inclusion of glycerol in formulations for pulmonary drug delivery may affect the bioavailability of inhaled steroids by retarding their transport across the lung epithelium. The aim of this study was to evaluate whether the molecular interactions of glycerol with model pulmonary interfaces provide a biophysical basis for glycerol modifying inhaled drug transport. Dipalmitoylphosphatidylcholine (DPPC) monolayers and liposomes were used as model pulmonary interfaces, in order to examine the effects of bulk glycerol (0-30% w/w) on their structures and dynamics using complementary biophysical measurements and molecular dynamics (MD) simulations. Glycerol was found to preferentially interact with the carbonyl groups in the interfacial region of DPPC and with phosphate and choline in the headgroup, thus causing an increase in the size of the headgroup solvation shell, as evidenced by an expansion of DPPC monolayers (molecular area increased from 52 to 68 Å2) and bilayers seen in both Langmuir isotherms and MD simulations. Both small angle neutron scattering and MD simulations indicated a reduction in gel phase DPPC bilayer thickness by ∼3 Å in 30% w/w glycerol, a phenomenon consistent with the observation from FTIR data, that glycerol caused the lipid headgroup to remain oriented parallel to the membrane plane in contrast to its more perpendicular conformation adopted in pure water. Furthermore, FTIR measurements suggested that the terminal methyl groups of the DPPC acyl chains were constrained in the presence of glycerol. This observation is supported by MD simulations, which predict bridging between adjacent DPPC headgroups by glycerol as a possible source of its putative membrane stiffening effect. Collectively, these data indicate that glycerol preferentially solvates DPPC headgroups and localizes in specific areas of the interfacial region, resulting in structural changes to DPPC bilayers which may influence cell permeability to drugs.


Novel Galectin-3 Roles in Neurogenesis, Inflammation and Neurological Diseases.
Galectin-3 (Gal-3) is an evolutionarily conserved and multifunctional protein that drives inflammation in disease. Gal-3's role in the central nervous system has been less studied than in the immune system. However, recent studies show it exacerbates Alzheimer's disease and is upregulated in a large variety of brain injuries, while loss of Gal-3 function can diminish symptoms of neurodegenerative diseases such as Alzheimer's. Several novel molecular pathways for Gal-3 were recently uncovered. It is a natural ligand for TREM2 (triggering receptor expressed on myeloid cells), TLR4 (Toll-like receptor 4), and IR (insulin receptor). Gal-3 regulates a number of pathways including stimulation of bone morphogenetic protein (BMP) signaling and modulating Wnt signalling in a context-dependent manner. Gal-3 typically acts in pathology but is now known to affect subventricular zone (SVZ) neurogenesis and gliogenesis in the healthy brain. Despite its myriad interactors, Gal-3 has surprisingly specific and important functions in regulating SVZ neurogenesis in disease. Gal-1, a similar lectin often co-expressed with Gal-3, also has profound effects on brain pathology and adult neurogenesis. Remarkably, Gal-3's carbohydrate recognition domain bears structural similarity to the SARS-CoV-2 virus spike protein necessary for cell entry. Gal-3 can be targeted pharmacologically and is a valid target for several diseases involving brain inflammation. The wealth of molecular pathways now known further suggest its modulation could be therapeutically useful.


Loss of inhibition in the spinal cord during barbiturate withdrawal.
Three types of inhibition in the spinal cord were studied during barbiturate withdrawal. Cats were made physically dependent on pentobarbital using a standard technique for chronic treatment developed in this laboratory. All animals treated this way developed severe withdrawal signs including spontaneous grand mal type convulsions, upon abrupt drug withdrawal. Animals were prepared for electrophysiological studies 48 hours after the last dose of chronic pentobarbital treatment, near peak intensity of the withdrawal syndrome. Under halothane anesthesia, the neuraxis was sectioned at C1 and both common carotid arteries were ligated. The animal was then respired on room air. The lumbar spinal cord was exposed by laminectomy, a hindleg was dissected to allow stimulation of peripheral nerves and action potentials were recorded from ventral roots L7 and S1 that had been cut near their exits through the dura mater. Recurrent (Renshaw cell-mediated) inhibition was almost totally absent during barbiturate withdrawal. Presynaptic inhibition was also decreased. Postsynaptic inhibition was little affected by barbiturate dependence. These findings indicate that loss of function in inhibitory pathways plays a role in barbiturate dependence and withdrawal.