Analyzing the Impact of Image Denoising and Segmentation on Melanoma Classification Using Convolutional Neural Networks.
Early skin cancer detection and its treatment are crucial for reducing death rates worldwide. Deep learning techniques have been used successfully to develop an automatic lesion detection system. This study explores the impact of pre-processing steps such as data augmentation, contrast enhancement, and segmentation on improving the convolutional neural network (CNN) performance for lesion classification. The classification network was designed from scratch by uniquely organizing its layers and using a different number of kernels, depth of the network, size, and hyperparameters. In addition, the network's performance was improved by pre-processing and segmentation steps. The proposed network was compared with the current state-of-the-art to demonstrate its best performance on the benchmark HAM10000 lesion dataset. The experimental study revealed that the classification network using denoised+segmented data achieved an accuracy (ACC), precision (PRE), recall (REC), specificity (SPE), and F-score of 93.40%, 93.45%, 94.51%, 92.08%, and 93.98%, respectively. To conclude, classification performance can be improved by incorporating pre-processing and segmentation steps.


Conformational composition, molecular structure and decomposition of difluorophosphoryl azide in the gas phase.
The conformational composition, molecular structure and decomposition of difluorophosphoryl azide F2P(O)N3 in the gas phase were studied using gas electron diffraction (GED), matrix isolation IR spectroscopy, and quantum-chemical calculations, respectively. While computational methods predict only minor differences in the total energy between the two possible conformers (syn and anti), the analysis of electron diffraction data reveals the dominating abundance of the syn conformer in the gas phase at room temperature. Ab initio frequency analyses suggest that a low-frequency large-amplitude motion of the N3 group with respect to the P-N-N-N torsion is to be expected for the syn conformer. The large amplitude motion was included explicitly into the GED structure refinement procedure. It presumably contributes to a thermodynamic stabilization of the syn-conformer with respect to the anti-conformer in the gas phase at ambient temperature. Upon flash vacuum pyrolysis, this syn conformer undergoes a stepwise decomposition via the difluorophosphoryl nitrene, F2P(O)N, which features as the first experimentally observed phosphoryl nitrene that can be thermally produced in the gas phase. To reveal the reaction mechanism, quantum-chemical calculations on the potential energy surface (PES) of F2P(O)N3 were performed. Both B3LYP/6-311+G(3df) and CBS-QB3 calculation results strongly support a stepwise decomposition into the singlet F2P(O)N, which prefers intersystem crossing to the thermally persistent triplet ground state instead of a Curtius rearrangement into FP(O)NF.


Differences in autonomic activity in individuals with optimal, normal, and high-normal blood pressure levels.
We investigated differences in autonomic activity in normotensive individuals having optimal, normal and high-normal blood pressure (BP) levels according to the guidelines of the European Society of Hypertension and European Society of Cardiology (ESH/ESC). The study included 294 normotensive subjects (135 males, 159 females; age range 16 to 75 years) with similar clinical, morphometric, biochemical, electrocardiographic, and echocardiographic features. The subjects were classified into the following BP groups: group 1 (n=113) with optimal BP (<120/80 mmHg); group 2 (n=104) with normal BP (120-129/80-84 mmHg), and group 3 (n=77) with high-normal BP (130-139/85-89 mmHg). All the subjects underwent 24-hour Holter monitoring to obtain heart rate variability (HRV) parameters of 24-hour, daytime, and nighttime periods. Normalized low (LFn) and high (HFn) frequency powers, and logarithmic (Log) values of HRV parameters were also calculated. On 24-hour Holter monitoring, heart rates were similar in three groups. Compared to group 1 and 2, group 3 exhibited significantly higher LF/HF (p<0.001) and LFn (p=0.001) values, and significantly lower HFn (p=0.001), pNN50 (p=0.001), and rMSSD (p=0.005) values. There were no significant differences between the groups with respect to daytime HRV parameters; however, nighttime LF/HF, LFn, and HFn values were significantly different between the groups. Log LF/HF values obtained during the 24-hour and nighttime periods showed significant differences between group 1 and group 3 (for 24 hours, p<0.001; for night, p=0.001) and between group 2 and group 3 (for 24 hours, p<0.001; for night, p=0.009), but group 1 and group 2 did not differ significantly in this respect (p>0.05). These findings suggest that subjects with high-normal BP have increased sympathetic activity and decreased parasympathetic activity, possibly making them more liable to hypertension.


[Hepatic lesion caused by halogenated anesthetics].
Two patients has been presented to whom general anaesthesia consecutive with halogenated anesthetics has been administrated. In one of the observations the first anaesthesia was administrated with enfluorane and halothane was for the others. The two patients inured with jaundice and in one of them accompanied with hepatic insufficiency; both of them had favourable evolution. The histology of one of them show us a lesion compatible with injury caused by halogenated anaesthesia. In one of the patients appears evidences of hypersensitivity. It is request the possibility that the enfluorane could be acting as a sensibilisant drug of the hepatic injury produced by the halothane. This experience suggest us the danger already described of the consecutive use at short intervals of halogenated anesthetics. So long as fat patients are more sensible to the toxic action of these anesthetics it is suggested prevent in their cases.


Efficacy and safety of twice-daily aclidinium bromide in COPD patients: the ATTAIN study.
The efficacy and safety of two doses of aclidinium bromide were evaluated in patients with moderate to severe chronic obstructive pulmonary disease (COPD). In this 24-week, double-blind trial, patients were randomised to twice-daily aclidinium (200 μg or 400 μg) or placebo. The primary efficacy end-point was change in trough forced expiratory volume in 1 s (FEV(1)) at week 24. Other end-points included peak FEV(1), health status (St George's Respiratory Questionnaire; SGRQ) and dyspnoea (Transitional Dyspnoea Index; TDI). Overall, 828 patients were randomised. At week 24, significant improvements from baseline were observed with aclidinium 200 μg and 400 μg versus placebo for trough FEV(1) (99 and 128 mL; both p<0.0001) and peak FEV(1) (185 and 209 mL; both p<0.0001). Peak FEV(1) improvements on day 1 were comparable with week 24. Aclidinium 200 μg and 400 μg produced significant improvements over placebo in baseline-adjusted mean SGRQ total score (-3.8 and -4.6 units; p<0.001 and p<0.0001) and TDI focal score (0.6 and 1.0 units; p<0.05 and p<0.001) at week 24. With both aclidinium doses, the incidence of anticholinergic adverse events was low, and similar to placebo. Twice-daily aclidinium significantly improved bronchodilation, health status and dyspnoea, and was well tolerated in patients with COPD.


Multi-unit floating alginate system: effect of additives on ciprofloxacin release.
In an attempt to fabricate floating beads of ciprofloxacin, drugloaded alginate beads were prepared by simultaneous external and internal gelation. The effect of blending of alginate with gellan, hydroxypropyl methylcellulose, starch, and chitosan on the bead properties were evaluated. Beads were spherical with incorporation efficiency in the range of 52.81 +/- 2.64 to 78.95 +/- 1.92%. Beads exhibited buoyancy over a period of 7-24 hr based on the formulation variables. In vitro release of ciprofloxacin from the alginate beads in simulated gastric fluid (SGF) (0.1 N HCl, pH 1.2), was influenced significantly (p < 0.001) by the properties and concentration of additives. Among the polymers incorporated into alginate beads. Hydroxy propyl methylcellulose (HPMC) provided an extended release over 7 hr. The drug release predominately followed Higuchi's square root model.


Risk of falls associated with antiepileptic drug use in ambulatory elderly populations: A systematic review.
Falls are a major cause of morbidity and mortality in older adults. About a third of those aged 65 years or older fall at least once each year, which can result in hospitalizations, hip fractures and nursing home admissions that incur high costs to individuals, families and society. The objective of this clinical review was to assess the risk of falls in ambulatory older adults who take antiepileptic drugs, medications that can increase fall risk and decrease bone density. PubMed, EMBASE, MEDLINE and the Cochrane Library electronic databases were searched from inception to July 2014. Case-control, quasi-experimental and observational design studies published in English that assessed quantifiable fall risk associated with antiepileptic drug use in ambulatory patient populations with a mean or median age of 65 years or older were eligible for inclusion. One author screened all titles and abstracts from the initial search. Two authors independently reviewed and abstracted data from full-text articles that met eligibility criteria. Searches yielded 399 unique articles, of which 7 met inclusion criteria-4 prospective or longitudinal cohort studies, 1 cohort study with a nested case-control, 1 cross-sectional survey and 1 retrospective cross-sectional database analysis. Studies that calculated the relative risk of falls associated with antiepileptic drug use reported a range of 1.29 to 1.62. Studies that reported odds ratios of falls associated with antiepileptic drug use ranged from 1.75 to 6.2 for 1 fall or at least 1 fall and from 2.56 to 7.1 for more frequent falls. Health care professionals should monitor older adults while they take antiepileptic drugs to balance the need for such pharmacotherapy against an increased risk of falling and injuries from falls.


Surface plasmon resonance imaging of limited glycoprotein samples.
A surface plasmon resonance imaging method has been developed for high throughput recognition and determination of low level glycoproteins with limited sample volume at least down to 50 nL. Chicken ovalbumin and immunoglobulin G were chosen as model compounds while bovine serum albumin and lysozyme were used as control. Each protein, at a concentration of 0.0080-1.0 mg mL(-1), was printed on one gold sensing film, and the films were simultaneously reacted with a probe solution and viewed using a laboratory-built surface plasmon resonance imaging system. The imaging signals were dependent on the concentration and the type of analyte, with a limit of detection down to at least 0.5 ng. The glycoproteins dotted at either 1.0 mg mL(-1) or 0.010 mg mL(-1) were easily differentiated from the non-glycoproteins by reaction with 200 nM concanavalin A (con A), giving a limit of recognition down also to 0.5 ng glycoprotein. This imaging method was hence considered a new tool for analyzing glycoproteins.


IoT Federated Blockchain Learning at the Edge.
IoT devices are sorely underutilized in the medical field, especially within machine learning for medicine, yet they offer unrivaled benefits. IoT devices are low cost, energy efficient, small and intelligent devices [1].In this paper, we propose a distributed federated learning framework for IoT devices, more specifically for IoMT (In-ternet of Medical Things), using blockchain to allow for a decentralized scheme improving privacy and efficiency over a centralized system; this allows us to move from the cloud based architectures, that are prevalent, to the edge.The system is designed for three paradigms: 1) Training neural networks on IoT devices to allow for collaborative training of a shared model whilst decoupling the learning from the dataset [2] to ensure privacy [3]. Training is performed in an online manner simultaneously amongst all participants, allowing for training of actual data that may not have been present in a dataset collected in the traditional way and dynamically adapt the system whilst it is being trained. 2) Training of an IoMT system in a fully private manner such as to mitigate the issue with confidentiality of medical data and to build robust, and potentially bespoke [4], models where not much, if any, data exists. 3) Distribution of the actual network training, something federated learning itself does not do, to allow hospitals, for example, to utilize their spare computing resources to train network models.


Pharmacological stabilization of intracranial aneurysms in mice: a feasibility study.
An increasing number of unruptured intracranial aneurysms are being detected, partly due to the increased use of brain imaging techniques. Pharmacological stabilization of aneurysms for the prevention of aneurysmal rupture could potentially be an attractive alternative approach to clipping or coiling in patients with unruptured intracranial aneurysms. We have developed a mouse model of intracranial aneurysm that recapitulates key features of intracranial aneurysms. In this model, subarachnoid hemorrhage from aneurysmal rupture causes neurological symptoms that can be easily detected by a simple neurological examination. Using this model, we tested whether anti-inflammatory agents such as tetracycline derivatives, or a selective inhibitor of matrix metalloproteinases-2 and -9 (SB-3CT), can prevent the rupture of intracranial aneurysms. Aneurysms were induced by a combination of induced hypertension and a single injection of elastase into the cerebrospinal fluid in mice. Treatment with minocycline, doxycycline, or SB-3CT was started 6 days after aneurysm induction. Aneurysmal rupture was detected by neurological symptoms and confirmed by the presence of intracranial aneurysms with subarachnoid hemorrhage. Minocycline and doxycycline significantly reduced rupture rates (vehicle versus doxycycline=80% versus 35%, P<0.05; vehicle versus minocycline=73% versus 24%, P<0.05) without affecting the overall incidence of aneurysms. However, SB-3CT did not affect the rupture rate (62% versus 55%, P=0.53). Our data established the feasibility of using a mouse model of intracranial aneurysm to test pharmacological stabilization of aneurysms. Tetracycline derivatives could be potentially effective in preventing aneurysmal rupture.


Experiences of traumatic events and associations with PTSD and depression development in urban health care-seeking women.
Posttraumatic stress disorder (PTSD) is an anxiety disorder that occurs after a traumatic event and has been linked to psychiatric and physical health declines. Rates of PTSD are far higher in individuals with low incomes and who reside in urban areas compared to the general population. In this study, 250 urban health care-seeking women were interviewed for a diagnosis of PTSD, major depressive disorder, and also the experience of traumatic events. Multivariate logistic regressions were used to determine the associations between traumatic events and PTSD development. Survival analysis was used to determine if PTSD developed from assaultive and nonassaultive events differed in symptom duration. Eighty-six percent of women reported at least one traumatic event, 14.8% of women were diagnosed with current PTSD, and 19.6% with past PTSD. More than half of women with PTSD had comorbid depression. Assaultive traumatic events were most predictive of PTSD development. More than two thirds of the women who developed PTSD developed chronic PTSD. Women who developed PTSD from assaultive events experienced PTSD for at least twice the duration of women who developed PTSD from nonassaultive events. In conclusion, PTSD was very prevalent in urban health care-seeking women. Assaultive violence was most predictive of PTSD development and also nonremittance.


A calcineurin- and NFAT-dependent pathway is involved in α-synuclein-induced degeneration of midbrain dopaminergic neurons.
Parkinson's disease (PD), the most common degenerative movement disorder, is caused by a preferential loss of midbrain dopaminergic (mDA) neurons. Both α-synuclein (α-syn) missense and multiplication mutations have been linked to PD. However, the underlying intracellular signalling transduction pathways of α-syn-mediated mDA neurodegeneration remain elusive. Here, we show that transgenic expression of PD-related human α-syn A53T missense mutation promoted calcineurin (CN) activity and the subsequent nuclear translocation of nuclear factor of activated T cells (NFATs) in mDA neurons. α-syn enhanced the phosphatase activity of CN in both cell-free assays and cell lines transfected with either human wild-type or A53T α-syn. Furthermore, overexpression of α-syn A53T mutation significantly increased the CN-dependent nuclear import of NFATc3 in the mDA neurons of transgenic mice. More importantly, a pharmacological inhibition of CN by cyclosporine A (CsA) ameliorated the α-syn-induced loss of mDA neurons. These findings demonstrate an active involvement of CN- and NFAT-mediated signalling pathway in α-syn-mediated degeneration of mDA neurons in PD.


Graft vs. host disease and graft vs. myeloma effect after non-myeloablative allogeneic transplantation.
For many years progress in event free and overall survival in patients diagnosed with multiple myeloma have been modest, however recently newer therapeutic options have become available and, for a small but increasing subset of patients, an "operational cure" can be offered. Although autologous transplantation is associated with a prolongation in event free and overall survival as compared to conventional chemotherapy, there is no plateau in the survival curves. By contrast, the use of allogeneic hematopoietic stem cells provides a tumor-free stem cell source and graft-vs.-myeloma activity leading to a higher frequency of long term survivors in molecular remission. Unfortunately, allogeneic transplantation has been associated with high transplant-related mortality (TRM). Non-myeloablative or reduced intensity conditioning (RIC) regimens, designed to be immunosuppressive rather than myeloablative, in an effort to reduce the toxicity and TRM associated with high dose chemotherapy, but maintaining the GVM effect, have been developed showing up to 90% overall response rate and low TRM. Interestingly, in a significant proportion of patients disease response is preceded by GVHD, suggesting a clear relationship between GVHD and graft vs. myeloma effect. Nevertheless these patients are at risk of developing life threatening complications and, on the contrary, some patients who reach disease control after GVHD and respond to GVHD therapy may finally relapse. Thus, efforts to separate GVM and GVHD are still required in order to improve the outcome of myeloma patients receiving allogeneic transplantation.


[Distribution of monoamines in the hypothalamus and hypophysis of carp, Cyprinus carpio].
Monoaminergic fibres exhibiting green fluorescence have been revealed by the Falck histochemical fluorescence technique in the area of the hypothalamic preoptic nuclei and nucleus lateralis tuberis of the carp. Monoaminergic cerebrospinal fluid-contacting nerve cells with an intensive greenish and yellowish fluorescence were observed in the walls of the lateral and posterior recesses of the third ventricle. Monoaminergic fibres are few in the pro-and meta-adenohypophysis. Yellowish-green fluorescent fibres seem to be much more frequent in the meta-adenohypophysis. Fluorescence of the fibres is unstabel and of a weak intensity.


Overexpression of the Na(+)/K(+)/Cl(-) cotransporter gene induces cell proliferation and phenotypic transformation in mouse fibroblasts.
Na(+)/K(+)/Cl(-) cotransporter activity is stimulated in early G(1) phase of the cell cycle and this stimulation was shown to be an essential event in fibroblast cell proliferation. In order to elucidate further the role of the Na(+)/K(+)/Cl(-) cotransporter in cell proliferation, we overexpressed the gene encoding the Na(+)/K(+)/Cl(-) cotransporter in mouse fibroblasts, and analyzed cellular phenotypic changes. Mouse Balb/c 3T3 cells were stably transfected with the cDNA of the shark rectal gland Na(+)/K(+)/Cl(-) cotransporter gene (NKCC1), and expressed in a mammalian vector under the cytomegalovirus promoter (Balb/c-NKCC1 cells). The transfected cells exhibited up to 10-fold greater bumetanide-sensitive Rb(+) influx compared to the control cells. The Balb/c-NKCC1 cells have acquired a typical transformation phenotype indicated by: (1) Loss of contact inhibition exhibited by growth to a higher cell density in confluent cultures, and formation of cell foci; (2) proliferation in low serum concentrations; and (3) formation of cell colonies in soft agar. The control cells transfected with the NKCC1 gene inserted in the opposite orientation in the vector retained their normal phenotype. Furthermore, the two specific inhibitors of the Na(+)/K(+)/Cl(-) cotransporter activity; bumetanide and furosemide inhibited the clonogenic efficiency in the NKCC1 transfected cells. These control experiments indicate that the apparent transformation phenotype acquired by the Balb/c-NKCC1 cells was not merely associated with the process of transfection and selecting for the neomycin-resistant clones, but rather with the overexpression of the Na(+)/K(+)/Cl(-) cotransporter gene. In order to ascertain that the regulated and normal expression of the Na(+)/K(+)/Cl(-) cotransporter control cell proliferation, the effect of bumetanide a specific inhibitor of the cotransporter, was tested on Balb/c 3T3 cell proliferation, induced by fibroblasts growth factor (FGF) and fetal calf serum (FCS). Bumetanide inhibited synchronized Balb/c 3T3 cell exit from the G(0)/G(1) arrest and entering S-phase. The inhibition was reversible, as removal of bumetanide completely released cell proliferation. Taken together, these results propose that the NKCC1 gene is involved in the control of normal cell proliferation, while its overexpression results in apparent cell transformation, in a manner similar to some protooncogenes.


The Interglomerular Circuit Potently Inhibits Olfactory Bulb Output Neurons by Both Direct and Indirect Pathways.
Sensory processing shapes our perception. In mammals, odor information is encoded by combinatorial activity patterns of olfactory bulb (OB) glomeruli. Glomeruli are richly interconnected by short axon cells (SACs), which form the interglomerular circuit (IGC). It is unclear how the IGC impacts OB output to downstream neural circuits. We combined in vitro and in vivo electrophysiology with optogenetics in mice and found the following: (1) the IGC potently and monosynaptically inhibits the OB output neurons mitral/tufted cells (MTCs) by GABA release from SACs: (2) gap junction-mediated electrical coupling is strong for the SAC→MTC synapse, but negligible for the SAC→ETC synapse; (3) brief IGC-mediated inhibition is temporally prolonged by the intrinsic properties of MTCs; and (4) sniff frequency IGC activation in vivo generates persistent MTC inhibition. These findings suggest that the temporal sequence of glomerular activation by sensory input determines which stimulus features are transmitted to downstream olfactory networks and those filtered by lateral inhibition. Odor identity is encoded by combinatorial patterns of activated glomeruli, the initial signal transformation site of the olfactory system. Lateral circuit processing among activated glomeruli modulates olfactory signal transformation before transmission to higher brain centers. Using a combination of in vitro and in vivo optogenetics, this work demonstrates that interglomerular circuitry produces potent inhibition of olfactory bulb output neurons via direct chemical and electrical synapses as well as by indirect pathways. The direct inhibitory synaptic input engages mitral cell intrinsic membrane properties to generate inhibition that outlasts the initial synaptic action.


Propranolol: A "Pick and Roll" Team Player in Benign Tumors and Cancer Therapies.
Research on cancer therapies focuses on processes such as angiogenesis, cell signaling, stemness, metastasis, and drug resistance and inflammation, all of which are influenced by the cellular and molecular microenvironment of the tumor. Different strategies, such as antibodies, small chemicals, hormones, cytokines, and, recently, gene editing techniques, have been tested to reduce the malignancy and generate a harmful microenvironment for the tumor. Few therapeutic agents have shown benefits when administered alone, but a few more have demonstrated clear improvement when administered in combination with other therapeutic molecules. In 2008 (and for the first time in the clinic), the therapeutic benefits of the β-adrenergic receptor antagonist, propranolol, were described in benign tumors, such as infantile hemangioma. Propranolol, initially prescribed for high blood pressure, irregular heart rate, essential tremor, and anxiety, has shown, in the last decade, increasing evidence of its antitumoral properties in more than a dozen different types of cancer. Moreover, the use of propranolol in combination therapies with other drugs has shown synergistic antitumor effects. This review highlights the clinical trials in which propranolol is taking part as adjuvant therapy at single administration or in combinatorial human trials, arising as a good pick and roll partner in anticancer strategies.


Suppression of p16 alleviates the senescence-associated secretory phenotype.
Oncogene-induced senescence (OIS) is characterized by increased expression of the cell cycle inhibitor p16, leading to a hallmark cell cycle arrest. Suppression of p16 in this context drives proliferation, senescence bypass, and contributes to tumorigenesis. OIS cells are also characterized by the expression and secretion of a widely variable group of factors collectively termed the senescence-associated secretory phenotype (SASP). The SASP can be both beneficial and detrimental and affects the microenvironment in a highly context-dependent manner. The relationship between p16 suppression and the SASP remains unclear. Here, we show that knockdown of p16 decreases expression of the SASP factors and pro-inflammatory cytokines IL6 and CXCL8 in multiple models, including OIS and DNA damage-induced senescence. Notably, this is uncoupled from the senescence-associated cell cycle arrest. Moreover, low p16 expression in both cancer cell lines and patient samples correspond to decreased SASP gene expression, suggesting this is a universal effect of loss of p16 expression. Together, our data suggest that p16 regulates SASP gene expression, which has implications for understanding how p16 modulates both the senescent and tumor microenvironment.


The prevalence of physical activity or inactivity in a rural African American community.
A modified version of the Minnesota Leisure Time Physical Activity Questionnaire was used to survey the adult residents of a rural African American community in North Carolina regarding the frequency, duration, and intensity of a variety of typical physical activities throughout the year. Variation in activity levels was analyzed with respect to age, gender, marital status, income, educational level, and smoking. Some seasonal differences in activity levels between men and women and between married and unmarried individuals were detected. One finding of significance was the total median amount of physical activity among this rural sample of 1500 minutes per week, 12 times the minimal recommendation. Advantages and disadvantages of this type of instrument and the importance of cultural aspects of physical activity are presented.


Quorum sensing signal production and inhibition by coral-associated vibrios.
Corals are inhabited by complex communities of microbes that affect their growth and survival. Several studies suggest that coral disease may be attributed to the success of vibrios in out-competing other bacteria in the mucus and tissues of corals. Vibrios utilize a variety of quorum sensing (QS) signal molecules to regulate processes that could be used to colonize corals during adverse environmental conditions. We therefore screened a range of Vibrios isolated from a variety of healthy and diseased corals, for the production of the QS signal molecules, N-acylhomoserine lactones (AHLs) and the AI-2 (autoinducer-2) small furanone signal molecule. All 29 strains examined activated the AI-2 biosensor, but only 17 activated an AHL biosensor. Using reverse phase thin-layer chromatography, we showed that the effect of temperature on AHL production varied considerably among the isolates. For the first time, the QS inhibition by Vibrio harveyi is reported. This only occurred at higher temperatures and does not appear to be due to degradation of AHLs. The large diversity of vibrios and the different effects of temperature on signal production may partly explain the complexity of coral-associated community changes in response to environmental factors.


Analogous mechanisms of resistance to benzothiazinones and dinitrobenzamides in Mycobacterium smegmatis.
Tuberculosis is still a leading cause of death worldwide. The selection and spread of Mycobacterium tuberculosis multidrug-resistant (MDR-TB) and extensively drug-resistant strains (XDR-TB) is a severe public health problem. Recently, two different classes of chemical series, the benzothiazinones (BTZ) and the dinitrobenzamide (DNB) derivatives have been found to be highly active against M. tuberculosis, including XDR-TB strains. The target of BTZs is DprE1 protein which works in concert with DprE2 to form the heteromeric decaprenylphosphoryl-β-D-ribose 2'-epimerase, involved in Decaprenyl-Phospho-Arabinose (DPA) biosynthesis. Interestingly, it has been shown that the DNBs block the same pathway thus suggesting that both drugs could share the same target. Moreover, in Mycobacterium smegmatis the overexpression of the NfnB nitroreductase led to the inactivation of the BTZs by reduction of a critical nitro-group to an amino-group. In this work several spontaneous M. smegmatis mutants resistant to DNBs were isolated. Sixteen mutants, showing high levels of DNB resistance, exhibited a mutation in the Cys394 of DprE1. Using fluorescence titration and mass spectrometry it has been possible to monitor the binding between DprE1 and DNBs, achieving direct evidence that MSMEG_6382 is the cellular target of DNBs in mycobacteria. Additionally, M. smegmatis mutants having low levels of resistance to DNBs harbor various mutations in MSMEG_6503 gene encoding the transcriptional repressor of the nitroreductase NfnB. By LC/MS analysis it has been demonstrated that NfnB is responsible for DNB inactivation. Taken together, our data demonstrate that both DNB and BTZ drugs share common resistance mechanisms in M. smegmatis.


MEDiastinal Irradiation and CArdio-Toxic Effects (MEDICATE): Exploring the Relationship between Cardiac Irradiation and High Sensitivity Troponins.
Radiation-induced heart disease is a late effect of cardiac irradiation and has been shown in patients with lymphoma and thoracic cancers. There is no established measurement tool to detect acute cardiac damage. However, high sensitivity troponin I and T (HsTnI and HsTnT) and echocardiograms have shown promise in some studies. A pilot trial was conducted to characterise whether these instruments may detect subclinical radiotherapy-induced cardiac damage. Eligible patients received high cardiac doses defined by either at least 30 Gy to 5% of cardiac volume or a mean dose of 4 Gy. HsTnI and HsTnT were measured before radiotherapy and after 2 and 4 weeks of radiotherapy; three-dimensional echocardiograms were completed before and 1 year after radiotherapy. Of 19 patients, the median 'mean left ventricular dose' was 3.1 Gy and the 'mean cardiac dose' was 8.6 Gy. Significant positive associations between HsTnI and HsTnT were observed at all time points, but there was no significant association with cardiac dose. The mean left ventricular dose and the maximum left ventricular dose were, however, associated with a decrease in ejection fraction (P = 0.054, 0.043) as well as an increase in left ventricular strain (P = 0.058). This study suggests that HsTnI and HsTnT are intimately related, but detection of acute cardiac damage was not shown, potentially due to limitations of these markers or low radiotherapy doses using conformal techniques. Our results also suggest subacute damage at 1 year may depend on the dose to the left ventricle. Further studies are needed, as identification of early damage could facilitate the ability to closely monitor and intervene in patients at risk for radiation-induced heart disease.


Asynchronous accumulation of lettuce infectious yellows virus RNAs 1 and 2 and identification of an RNA 1 trans enhancer of RNA 2 accumulation.
Time course and mutational analyses were used to examine the accumulation in protoplasts of progeny RNAs of the bipartite Crinivirus, Lettuce infectious yellow virus (LIYV; family Closteroviridae). Hybridization analyses showed that simultaneous inoculation of LIYV RNAs 1 and 2 resulted in asynchronous accumulation of progeny LIYV RNAs. LIYV RNA 1 progeny genomic and subgenomic RNAs could be detected in protoplasts as early as 12 h postinoculation (p.i.) and accumulated to high levels by 24 h p.i. The LIYV RNA 1 open reading frame 2 (ORF 2) subgenomic RNA was the most abundant of all LIYV RNAs detected. In contrast, RNA 2 progeny were not readily detected until ca. 36 h p.i. Mutational analyses showed that in-frame stop codons introduced into five of seven RNA 2 ORFs did not affect accumulation of progeny LIYV RNA 1 or RNA 2, confirming that RNA 2 does not encode proteins necessary for LIYV RNA replication. Mutational analyses also supported that LIYV RNA 1 encodes proteins necessary for replication of LIYV RNAs 1 and 2. A mutation introduced into the LIYV RNA 1 region encoding the overlapping ORF 1B and ORF 2 was lethal. However, mutations introduced into only LIYV RNA 1 ORF 2 resulted in accumulation of progeny RNA 1 near or equal to wild-type RNA 1. In contrast, the RNA 1 ORF 2 mutants did not efficiently support the trans accumulation of LIYV RNA 2. Three distinct RNA 1 ORF 2 mutants were analyzed and all exhibited a similar phenotype for progeny LIYV RNA accumulation. These data suggest that the LIYV RNA 1 ORF 2 encodes a trans enhancer for RNA 2 accumulation.


[Primary treatment of ovarian cancer].
Patients with epithelial ovarian cancer are often diagnosed with advanced disease; hence they have a generally poor survival rate. The main objective was to assess the clinical effectiveness of the four main treatment options in the primary treatment of epithelial ovarian cancer: i) adjuvant chemotherapy and/or adjuvant radiotherapy, ii) cytoreductive surgery, iii) neoadjuvant chemotherapy in advanced disease, and iv) postoperative chemotherapy in advanced disease. The scientific literature was identified by searches in Medline, Embase and Cochrane CCTR and by additional manual searches. Using criteria defined by protocol, two reviewers assessed each study according to relevance, quality and validity. Among 2227 publications identified, 635 were read as full-text articles, and 90 studies were critically assessed as relevant publications. All included studies were systematised in three subgroups according to the quality of the study design in question and the validity of the results: high, moderate, or low. A total of 45 studies of high or moderate quality form the documentary basis for this review. Current data are inconclusive regarding the effect of adjuvant chemotherapy. Retrospective data show a survival advantage for patients who have had maximum cytoreductive surgery. The effect of neoadjuvant chemotherapy is uncertain. Several questions remain unanswered despite wide acceptance of paclitaxel-carboplatin as the current standard in first-line treatment. This systematic review demonstrates the need for more clinical studies.


Loss of myoepithelial calponin-1 characterizes high-risk ductal carcinoma in situ cases, which are further stratified by T cell composition.
A hallmark of ductal carcinoma in situ (DCIS) progression is a loss of the surrounding ductal myoepithelium. However, whether compromise in myoepithelial differentiation, rather than overt cellular loss, can be used to predict the risk of DCIS progression is unknown. Here we address this question utilizing pure and mixed DCIS cases (N = 30) as surrogates for DCIS at low and high risk for progression, respectively. We used multiplex immunohistochemical staining to evaluate the relationship between myoepithelial cell differentiation and lymphoid immune cell types associated with poor prognostic DCIS. Our results show that myoepithelial calponin-1 discriminates between pure and mixed DCIS lesions better than histological subtype, presence of necrosis, or nuclear grade. Additionally, focal loss of myoepithelial cells associated with increased PD-1+CD8+ T cells, which suggests a link between the myoepithelium and immune surveillance. To identify associations between calponin-1 expression and immune response, we performed unsupervised hierarchical clustering of myoepithelial and immune cell biomarkers on 219 DCIS lesions from 30 cases. Notably, the majority of pure (low-risk) DCIS lesions clustered in a high calponin-1, T cell low group, whereas the majority of mixed (high-risk) DCIS lesions clustered in a low calponin-1, T cell high group, specifically with CD8+ and PD-1+CD8+ T cells. However, a subset of pure DCIS lesions had a similar calponin-1 and immune signature as the majority of mixed DCIS lesions, which have low calponin-1 and T cell enrichment-raising the possibility that these pure DCIS lesions might be at a high risk for progression.


Elevated Behavioral Economic Demand for Alcohol in Co-Users of Alcohol and Cannabis.
Co-use of cannabis and alcohol is associated with increased drinking and other negative consequences relative to use of alcohol alone. One potential explanation for these differences is overvaluation of alcohol (e.g., alcohol demand) among co-users, similar to established overvaluation of alcohol among tobacco and alcohol co-users. This study examined differences in alcohol demand between an alcohol and cannabis co-user group and an alcohol-only group. A large sample of adult drinkers (n = 1,643, 54% female) was recruited through an online crowdsourcing site (Amazon Mechanical Turk). Of the full sample, 476 participants reported weekly or greater cannabis use in the past 6 months (co-user group); 888 reported never using cannabis in the past 6 months (alcohol-only group). Assessments included a validated alcohol purchase task and self-report measures of alcohol and cannabis use. Co-users reported significantly higher alcohol consumption across the elastic portion of the alcohol demand curve (i.e., $1.50-$9.00/drink). Analyses of covariance controlling for alcohol use and demographics revealed significantly higher breakpoint (p = .025) and Omax (p = .002) and significantly lower elasticity (p < .003) in the co-user group. Intensity and Pmax did not significantly differ between groups. Co-users of cannabis and alcohol overvalue alcohol compared with individuals who drink alcohol but do not use cannabis. This study is generally consistent with prior studies on alcohol and tobacco co-users, providing converging evidence that polysubstance use is associated with overvaluation of alcohol. These findings have important implications for treatment and prevention, particularly in the context of changes in cannabis legalization.


Anthelminthic activity of the cyclotides (kalata B1 and B2) against schistosome parasites.
The risk of reduced sensitivity of the human schistosomes to praziquantel has led to efforts to find new therapies. Here, the cyclotides kalata B1 (kB1), kalata B2 (kB2), MCoCC-1, and MCoTI-II, cyclic peptides extracted from plants and shown to be potent against nematodes and insects, were tested for antischistosome activity. In vitro assays showed that high concentrations (500-1000 μg/mL) of either kB1 or kB2 killed Schistosoma japonicum and Schistosoma mansoni adults within 5 min, whereas MCoTI-II and MCoCC-1 had no effect. Lethal concentrations to kill 50% of the population for kB2 was 15.5 ± 7.4 μg/mL at 1 h for male S. japonicum (Philippine strain). Males were more susceptible than females. kB2 showed higher antischistosome activity than kB1 and killing time was concentration-dependent. Mode of action studies revealed that kB1 and 2 lysed the tegument of adult worms. Lysis of myofibrils was not demonstrated, but longitudinal and radial muscle fibers were distorted, an observation consistent with strong coiling of the parasites after drug exposure. A single dose of kB2 administered either orally or intravenously, reduced worm burdens in S. japonicum-infected mice from 15% to 60%. However, treatment of S. mansoni-infected mice did not result in reduction in worm burdens. Our studies show that kB2 acts as a promising antischistosomal against Philippine S. japonicum, and it or other cyclotides may be developed further as general anthelminthics. With thousands of cyclotides predicted to occur in plants, and the amenability of these peptides to combinatorial variation, there is potential for their exploitation as wide-spectrum anthelminthics.


Amyloid myopathy: an underdiagnosed entity.
Amyloidosis can involve multiple organs, including kidney, heart, peripheral nerve, skin, joints, and skeletal muscle, but rarely presents as a myopathy. We studied 13 adults with muscle weakness for between 3 months and 4 years in whom the diagnosis of systemic amyloidosis was unsuspected before or until just before the time of the muscle biopsy. All muscle specimens demonstrated congophilic deposits around blood vessels and muscle fibers, some necrotic and regenerating fibers, and signs of mild denervation. Immunostains in 10 patients revealed immunoglobulin amyloidosis in 7 and gelsolin amyloidosis in 1. Apolipoprotein E co-localized with the congophilic deposits in all 10, and a C-terminal epitope of the beta-amyloid precursor protein was detected in 6. The frequency of the diagnosis of amyloid myopathy increased 10-fold when we adopted the fluorescent Congo red stain as a routine procedure in assessing muscle biopsy specimens.


Physiologic changes of compound muscle action potentials related to voluntary contraction and muscle length in carpal tunnel syndrome.
The affect of muscle length and voluntary contraction upon compound muscle action potentials (CMAPs) in subjects with carpal tunnel syndrome (CTS) has been evaluated. Twenty-five hands in a CTS patient group and 29 hands in a normal subject control group were studied. The CMAPs from the abductor pollicis brevis induced by median nerve stimulation at the wrist were obtained for five thumb positions: neutral, abduction for shortening with and without contraction, and adduction for lengthening with and without contraction. Upon muscle shortening with relaxation, CMAP duration decreased in both groups, whereas waveform amplitude increased in the control group and showed no significant change in the CTS group. Muscle shortening with contraction afforded decreased CMAP duration and increased CMAP amplitude in both groups. Upon muscle lengthening with relaxation, both groups showed a reduction in CMAP amplitude and an increase in CMAP duration. Upon lengthening with contraction, CMAP duration decreased in the control group; in contrast, the CTS group showed further amplitude reduction and the waveform duration returned to the neutral value. These results demonstrate that, in patients with CTS, physiologic CMAP summations by muscle shortening or contraction may be less effective, whereas decreases in amplitude and increases in duration may be accentuated by lengthening and contraction.


Using a theoretical framework to develop postgraduate health professions education research and practice.
This article was migrated. The article was marked as recommended. Healthcare education is complex and multifaceted, requiring study from different angles and with different lenses. We propose that the use of a meta-framework can help those teaching and researching postgraduate health professions education make holistic sense of their practice and findings from different projects. We discuss how we have employed Bronfenbrenner's ecological systems theory (EST) as an overarching theoretical framework for the scholarship of learning and teaching in the context of postgraduate health professions education. Taking a structured approach to pedagogical thinking and research through the use of a meta-framework opens up useful ways of framing findings and further questions, locating research projects within a bigger picture, and communicating to others the focus of a research programme. We address the problem of the under-theorizing of educational research in postgraduate health professions education, advocating both theoretical frameworks for individual research projects, and an overarching theoretical "meta-framework" to interrogate and draw together multiple studies. In doing so we build on, critique and further develop Bronfenbrenner's ecological systems theory.


Development of a new mouth opening force test using an indirect cervical traction device.
There have been few reports of objective jaw opening tests, and such studies have involved devices specifically designed for research. In the present study, in order to conveniently and objectively assess mouth opening movements, we replaced the manual resistance used in Daniels and Worthingham's muscle test (DMT) with an indirect cervical traction device. We examined whether the maximum mouth opening force (MOF) could be reliably quantified using this device. The subjects were 12 healthy individuals with a mean age of 28.8 years. The MOF measurement procedure was as follows: 1) the subject sat in the chair, and a head belt was placed under the chin so that a traction force was applied almost parallel to the body axis; 2) the researcher instructed the subject to maintain the maximum mouth opening; 3) as maximum resistance was approached, the rate of increase in the traction force decreased. Maximum opening force was recorded; 4) one measurement was taken per session, for a total of two measurements per subject. Pearson's correlation coefficients were used to assess the reproducibility of MOF values. The average MOF (mean +/- SD) in the first and second tests was 24.2 +/- 1.9 and 24.5 +/- 2.0 kg. There was an extremely high correlation between first and second measurements (r = 0.969). The presently described indirect cervical traction device can be used to reliably quantify MOF.


Patient-reported outcome measures in the NHS: new methods for analysing and reporting EQ-5D data.
In a landmark move, the UK Department of Health (DH) has introduced the routine collection of patient-reported outcome measures (PROMs) to measure the performance of health-care providers. From April 2009, generic (EQ-5D) and condition-specific PROMs are being collected from patients before and after four surgical procedures; eventually this will be extended to include a wide range of other NHS services. The aim of this article is to report analysis of the EQ-5D data generated from a pilot study commissioned by the DH and to consider the implications for the use of EQ-5D data in performance indicators and measures of patient benefit. We present two new methods that we have developed for analysing and displaying EQ-5D profile data: a Paretian Classification of Health Change and a health profile grid. We show that EQ-5D profile data can be readily analysed to generate insights into the nature of changes in patient-reported health that would be obscured by summarising these profiles by their index scores, or focusing just on the post operative outcomes. Our methods indicate differences between providers and between sub-groups of patients. Our results also show striking differences in changes in EQ-5D profiles between surgical procedures, which require further investigation.


Protective effects of deep sea water against doxorubicin‑induced cardiotoxicity in H9c2 cardiac muscle cells.
Doxorubicin (DOX) is one of the most effective chemotherapeutic agents in the treatment of a variety of tumors. However, its clinical use has been compromised by the risk of cardiotoxicity. Thus, many efforts have been focused on exploring new strategies to prevent or reverse DOX-induced cardiotoxicity. Recently, deep sea water (DSW) has drawn much scientific interest for therapeutic intervention due to its enrichment in nutrients and minerals. In this study, we investigated whether DSW has protective effects against DOX-induced cardiotoxicity. Pre-treatment with DSW significantly increased the viability of DOX-treated rat H9c2 cardiac muscle cells. This protective effect of DSW appears to be mediated through the inhibition of DNA damage rather than suppression of reactive oxygen species (ROS) production in DOX‑treated H9c2 cardiac muscle cells. The inhibitory effect of DSW on DOX-induced DNA damage subsequently attenuated apoptotic signaling such as activation of cysteine-aspartic acid protease-3 (caspase-3) and fragmentation of poly(ADP-ribose) polymerase (PARP), whereas the expression of anti-apoptotic protein B-cell lymphoma-extra large (Bcl-xL) was increased. Moreover, DSW treatment rescued the activation of protein kinase B (Akt) to protect cells from DOX-triggered apoptosis. Taken together, our data showed that DSW has protective effects against DOX-induced cardiotoxicity, suggesting that DSW has some promise as a novel protective supplement for promoting the successful use of DOX in clinical regimen.


MR_CHIROD v.2: magnetic resonance compatible smart hand rehabilitation device for brain imaging.
This paper presents the design, fabrication, and testing of a novel, one degree-of-freedom, magnetic resonance compatible smart hand interfaced rehabilitation device (MR_CHIROD v.2), which may be used in brain magnetic resonance (MR) imaging during handgrip rehabilitation. A key feature of the device is the use of electrorheological fluids (ERFs) to achieve computer controlled, variable, and tunable resistive force generation. The device consists of three major subsystems: 1) an ERF based resistive element, 2) handles, and c) two sensors, one optical encoder and one force sensor, to measure the patient induced motion and force. MR_CHIROD v.2 is designed to resist up to 50% of the maximum level of gripping force of a human hand and be controlled in real time. Our results demonstrate that the MR environment does not interfere with the performance of the MR_CHIROD v.2, and, reciprocally, its use does not cause fMR image artifacts. The results are encouraging in jointly using MR_CHIROD v.2 and brain MR imaging to study motor performance and assess rehabilitation after neurological injuries such as stroke.


Insulin/IGF-I binding ratio in skeletal and cardiac muscles of vertebrates: a phylogenetic approach.
Insulin and insulin-like growth factor (IGF-I) receptor binding and tyrosine kinase activity were characterized in cardiac and skeletal muscles of several vertebrates. Specific insulin binding per unit weight of skeletal muscle was clearly higher in pigeon and rat than in ectothermic vertebrates (32 +/- 5 and 25 +/- 2.7%/100 mg initial tissue in pigeon and rat, respectively, vs. 4.4 +/- 0.2%/100 mg in carp samples). Insulin binding clearly predominated over IGF-I binding in skeletal muscle of endotherms (IGF-I binding was 7.7 +/- 0.5%/100 mg in rat). In ectothermic vertebrates the situation was reversed, and IGF-I binding was higher than insulin binding. In cardiac muscle, specific binding of both insulin and especially IGF-I was higher than the values found in skeletal muscle of the same species (IGF-I binding was 60 +/- 4, 103 +/- 2, and 20 +/- 3%/100 mg in carp, turtle, and rat, respectively). The tyrosine kinase activity of insulin and IGF-I receptors of all species studied presented basal phosphotransferase rates (250-1,600 fmol P.micrograms protein-1.30 min-1) and percentage of stimulation (150-520%) with clear differences between species. The present data suggest that insulin and IGF-I binding to skeletal and cardiac muscles change through the vertebrate scale in both quantity and activity.


Correlation between anti-mutated citrullinated vimentin and bone turnover markers (CTX-1 and P1NP) in patients with rheumatoid arthritis in remission and low-disease activity.
Rheumatoid arthritis is associated with bone loss and the risk of osteoporotic fracture. Bone loss in this disease is mediated by inflammation and autoimmunity. Many studies have shown that anti-citrullinated protein antibody is capable of inducing bone loss through several mechanisms.This study aimed to determine the relationship between autoimmunity, represented by anti-mutated citrullinated vimentin (anti-MCV) in this study, and bone loss, represented by C-terminal cross-linking telopeptide of type I collagen (CTX-1), and N-terminal pro-peptide of type 1 procollagen (P1NP) in this study, in patients with rheumatoid arthritis in remission and low disease activity. This study enrolled 38 rheumatoid arthritis patients with disease remission and low disease activity in Cipto Mangunkusumo Hospital between August and September 2019. We collected the patients' demographic data, Disease Activity Score 28 (DAS28), and treatment history. All participants underwent blood work for anti-MCV, CTX-1, and P1NP. Thirty-four of the subjects were women (89.5%), with the mean age of 40 ±7.6 years and the median disease duration of 36 months. Among the subjects, 26 patients (68.4%) were anti-MCV positive. There was no correlation between anti-MCV and CTX-1 levels (r = 0.101, p = 0.274). There was a moderate negative correlation between anti-MCV and P1NP (r = -0.449, p = 0.001). The mean difference of P1NP according to anti-MCV level also showed a significant difference (p = 0.019). The anti-MCV levels are not directly correlated with CTX-1 levels, indicating heterogeneity in the disease course even after inflammation has ceased. The anti-MCV and P1NP levels are moderately correlated, indicating that bone formation is resumed during the suppression of autoimmunity.


A case-control study of non-Hodgkin's lymphoma and the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) in eastern Nebraska.
To evaluate the role of the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) in the development of non-Hodgkin's lymphoma (NHL), we conducted a population-based, case-control study in 66 counties in eastern Nebraska. Telephone interviews were conducted with 201 white men diagnosed with NHL between July 1, 1983, and June 30, 1986, and with 725 controls. There was a 50% excess of NHL among men who mixed or applied 2,4-D (odds ratio [OR] = 1.5; 95% confidence interval = 0.9, 2.5). The risk of NHL increased with the average frequency of use to over threefold for those exposed 20 or more days per year (p for trend = 0.051). Adjusting for use of organophosphate insecticides lowered the risk estimate for frequent users (OR = 1.8), but adjustment for fungicide use increased the risk estimate (OR = 4.5). Simultaneous adjustment for organophosphates and fungicides yielded an OR of 3.1 for farmers who mixed or applied 2,4-D more than 20 days per year. Risk also increased with degree of exposure, as indicated by application method and time spent in contaminated clothing, but not with the number of years of 2,4-D use or failure to use protective equipment. Although other pesticides, especially organophosphate insecticides, may be related to NHL, the risk associated with 2,4-D does not appear to be explained completely by these other exposures.


Serum somatomedin C concentrations following pneumonectomy.
Because a somatomedinlike substance appears in rabbit serum 9 days after pneumonectomy, which has the capacity to make cultured type II cells incorporate 3H thymidine into DNA, we measured serum immunoreactive somatomedin C concentrations following pneumonectomy, nephrectomy, and sham operation (thoracotomy). We found no striking differences in somatomedin C levels between the 3 groups at 5, 9, 13, 17, and 21 days postsurgery, indicating that somatomedin-C and the serum postpneumonectomy factor are unlikely to be the same. Serum somatomedin-C concentrations, however, rose following surgery in all groups, suggesting that serum somatomedin-C responds to surgical wounding. We speculate that the increase in serum somatomedin-C reflects production of somatomedin-C by the healing tissue.


Characterization of pseudorabies virus antibody responses in young swine after infection and vaccination by using an immunoglobulin M antibody capture enzyme-linked immunosorbent assay.
An immunoglobulin M (IgM) antibody capture enzyme-linked immunosorbent assay (MACELISA) was developed for the detection of pseudorabies virus (PRV)-specific IgM antibody in swine sera because false-positive reactions frequently occurred when sera from older swine were tested with an indirect IgM enzyme-linked immunosorbent assay. Monoclonal mouse anti-swine IgM was used as the capturing antibody, and rabbit anti-PRV hyperimmune gamma globulin was used as the indicating antibody. Sera from non-PRV-infected, experimentally infected, vaccinated and challenged, passively immune and challenged, and naturally infected swine were evaluated. The PRV MACELISA had a specificity of 95% and was as sensitive and reproducible as previously reported in direct assays. An antibody response was still detectable with the MACELISA 21 days after inoculation. The PRV MACELISA did not detect a consistent antibody response in sera from swine vaccinated with either killed-PRV or modified live-virus vaccines but did detect an antibody response in sera from passively immune pigs after challenge with virulent PRV. These results indicated that the PRV MACELISA may be useful for the rapid serodiagnosis of recent PRV infection in swine.


Relationship of HLA antigens and cryoglobulinaemia in hepatitis C virus infected patients.
To find the relationship between human leukocyte antigens (HLA) and cryoglobulin positivity in hepatitis C virus (HCV) infected individuals. Eligible individuals selected from pre and post renal transplant settings were divided into three groups. Group A (n = 301) consisted of normal controls, while group B (n = 200) comprised of pathological controls that were HCV antibody (anti-HCV) positive but negative for cryoglobulins. Group C comprised of 56 anti-HCV positive, cryoglobulin positive patients. HLA-A, -B and -DRB1 loci were typed by polymerase chain reaction (PCR) method and relationship between HLA antigens, anti-HCV status and cryoglobulinaemia was analyzed. HLA-A*02, -B*57 and -DRB1*03 were more frequently found among group C members as compared to groups A and B. Only HLA-B* 57 occurrence reached statistical significance (14.3% versus 6% and 4%, corrected P-value = 0.045 and 0.012 and OR = 2.6 and 4 respectively) No differences in the distribution of HLA antigens were seen among healthy and pathological controls. The presence of HLA-B*57 confers susceptibility to cryoglobulinaemia in HCV infected patients in our population. HCV positive renal transplant recipients with these alleles should be monitored for cryoglobulin formation).


Piecemeal recruitment of left-lateralized brain areas during reading: a spatio-functional account.
Neuroimaging studies of reading converge to suggest that linguistically elementary stimuli are confined to the activation of bilateral posterior regions, whereas linguistically complex stimuli additionally recruit left hemispheric anterior regions, raising the hypotheses of a gradual bilateral-to-left and a posterior-to-anterior recruitment of reading related areas. Here, we tested these two hypotheses by contrasting a repertoire of eight categories of stimuli ranging from simple orthographic-like characters to words and pseudowords in a single experiment, and by measuring BOLD signal changes and connectivity while 16 fluent readers passively viewed the stimuli. Our results confirm the existence of a bilateral-to-left and posterior-to-anterior recruitment of reading related areas, straightforwardly resulting from the increase in stimuli's linguistic processing load, which reflects reading processes: visual analysis, orthographic encoding and phonological decoding. Connectivity analyses strengthened the validity of these observations and additionally revealed an enhancement of the left parieto-frontal information trafficking for higher linguistic processing. Our findings clearly establish the notion of a gradual spatio-functional recruitment of reading areas and demonstrate, to the best of our knowledge, the first evidence of a robust and staged link between the level of linguistic processing, the spatial distribution of brain activity and its information trafficking.


ASTROPHYSICS: Neutron Stars Imply Relativity's a Drag.
A new finding, based on x-rays from distant neutron stars, could be the first clear evidence of a weird relativistic effect called frame dragging, in which a heavy chunk of spinning matter wrenches the space-time around it like an eggbeater. Using data from NASA's Rossi X-ray Timing Explorer, three astronomers in Amsterdam found circumstantial evidence for frame dragging in the flickering of three neutron stars in binary systems. They announced their results in the 1 September issue of The Astrophysical Journal.


mtDNA-Server: next-generation sequencing data analysis of human mitochondrial DNA in the cloud.
Next generation sequencing (NGS) allows investigating mitochondrial DNA (mtDNA) characteristics such as heteroplasmy (i.e. intra-individual sequence variation) to a higher level of detail. While several pipelines for analyzing heteroplasmies exist, issues in usability, accuracy of results and interpreting final data limit their usage. Here we present mtDNA-Server, a scalable web server for the analysis of mtDNA studies of any size with a special focus on usability as well as reliable identification and quantification of heteroplasmic variants. The mtDNA-Server workflow includes parallel read alignment, heteroplasmy detection, artefact or contamination identification, variant annotation as well as several quality control metrics, often neglected in current mtDNA NGS studies. All computational steps are parallelized with Hadoop MapReduce and executed graphically with Cloudgene. We validated the underlying heteroplasmy and contamination detection model by generating four artificial sample mix-ups on two different NGS devices. Our evaluation data shows that mtDNA-Server detects heteroplasmies and artificial recombinations down to the 1% level with perfect specificity and outperforms existing approaches regarding sensitivity. mtDNA-Server is currently able to analyze the 1000G Phase 3 data (n = 2,504) in less than 5 h and is freely accessible at https://mtdna-server.uibk.ac.at.


Impact of introducing the International Association of Diabetes and Pregnancy Study Groups (IADPSG) criteria on pregnancy outcomes in Japan.
To estimate pregnancy complications in women newly diagnosed with gestational diabetes mellitus (GDM) according to the new International Association of Diabetes and Pregnancy Study Groups (IADPSG) criteria and verify the validity of introducing the IADPSG criteria in Japan. This retrospective study included data from women with singleton delivery at 22 weeks or later at a tertiary hospital during 2000-2009. We compared pregnancy outcomes between women who would now be diagnosed with GDM according to the IADPSG criteria but not by the old JSOG criteria (IGT group, n = 503) and women with normal glucose tolerance according to both the criteria (NGT group, n = 2,789). Multivariate analysis was performed and adjusted for background factors. Maternal age at delivery and pre-pregnancy BMI were significantly higher in the IGT group than in the NGT group, while gestational weeks at delivery did not differ between the groups. No difference was observed in the rates of GDM-related composite complications (defined as cases with at least one of the following: macrosomia, shoulder dystocia, neonatal hypoglycemia, neonatal hyperbilirubinemia, or neonatal respiratory distress syndrome) at 11.9% and 8.8% (adjusted odds ratio (OR) 1.30, 95% confidence interval (CI) 0.90-1.87, p = 0.16). Pregnancy outcomes did not differ significantly between the IGT and NGT groups, except for frequencies of total neonatal admissions at 10.5% and 7.1%, respectively (adjusted OR 1.55, 95% CI 1.12-2.13, p < 0.01).


Multi-level protection of rights: India's Mental Healthcare Act, 2017 and the United Nations' Convention on the Rights of Persons with Disabilities (Articles 12 and 14).
Mental health law is a feature of mental health systems around the world, so legislative reform is an important way to advance compliance with the United Nations' Convention on the Rights of Persons with Disabilities (CRPD). This paper provides an overview of India's Mental Healthcare Act, 2017 in the context of Articles 12 ('Equal recognition before the law') and 14 ('Liberty and security of person'). Notwithstanding the potential of law to produce change, exclusive or disproportionate emphasis on law draws attention from other areas in need of equal if not greater reform, such as public education, mental health services, physical healthcare, social care networks, criminal justice systems, and gathering political backing for the profound reforms needed to transform the socio-economic landscapes in which mental illness develops, diagnoses are made, treatment is provided (or not provided), recovery occurs (or does not occur), and social citizenship is protected and promoted (or eroded and obstructed). There is a particular issue with misunderstandings of psychiatry in academic discussions of mental health legislation which appears to stem from lack of engagement with the full diversity of service-user views and the evidence base for psychiatric care. Greater recognition of upstream factors that shape both suffering and services would acknowledge that the decisions of politicians and service managers have a much greater impact on rights, especially the right to treatment and support, than the decisions of individual clinicians whose options are often very limited at the level of individual care. India's legislation seeks to address not only individual-level care, but also this higher level of reform through its articulation of an explicit 'right to access mental healthcare and treatment'. Similar multi-level legislative reform elsewhere would increase cooperation from all stakeholders, improve compliance with the CRPD, and help move towards zero coercion in mental health services.


The role of the natural environment in the emergence of antibiotic resistance in gram-negative bacteria.
During the past 10 years, multidrug-resistant Gram-negative Enterobacteriaceae have become a substantial challenge to infection control. It has been suggested by clinicians that the effectiveness of antibiotics is in such rapid decline that, depending on the pathogen concerned, their future utility can be measured in decades or even years. Unless the rise in antibiotic resistance can be reversed, we can expect to see a substantial rise in incurable infection and fatality in both developed and developing regions. Antibiotic resistance develops through complex interactions, with resistance arising by de-novo mutation under clinical antibiotic selection or frequently by acquisition of mobile genes that have evolved over time in bacteria in the environment. The reservoir of resistance genes in the environment is due to a mix of naturally occurring resistance and those present in animal and human waste and the selective effects of pollutants, which can co-select for mobile genetic elements carrying multiple resistant genes. Less attention has been given to how anthropogenic activity might be causing evolution of antibiotic resistance in the environment. Although the economics of the pharmaceutical industry continue to restrict investment in novel biomedical responses, action must be taken to avoid the conjunction of factors that promote evolution and spread of antibiotic resistance.


Parkour LIMS: high-quality sample preparation in next generation sequencing.
This paper presents Parkour, a software package for sample processing and quality management of next generation sequencing data and samples. Starting with user requests, Parkour allows tracking and assessing samples based on predefined quality criteria through different stages of the sample preparation workflow. Ideally suited for academic core laboratories, the software aims to maximize efficiency and reduce turnaround time by intelligent sample grouping and a clear assignment of staff to work units. Tools for automated invoicing, interactive statistics on facility usage and simple report generation minimize administrative tasks. Provided as a web application, Parkour is a convenient tool for both deep sequencing service users and laboratory personal. A set of web APIs allow coordinated information sharing with local and remote bioinformaticians. The flexible structure allows workflow customization and simple addition of new features as well as the expansion to other domains. The code and documentation are available at https://github.com/maxplanck-ie/parkour. Supplementary data are available at Bioinformatics online.


Study on the possible association of brain-derived neurotrophic factor polymorphism with the developmental course of symptoms of attention deficit and hyperactivity.
Several studies have, with conflicting results, investigated the relationship between the Val⁶⁶Met polymorphism in brain-derived neurotrophic factor (BDNF) and attention deficit hyperactivity disorder (ADHD). We assessed longitudinal, quantitative phenotypes of hyperactivity-impulsivity and inattention in order to determine whether the Val⁶⁶Met polymorphism is associated with age-specific and/or persistent symptoms of hyperactivity-impulsivity and/or inattention in a community-based cohort of 1236 Swedish individuals for which ADHD symptom data were collected when the participants were aged 8-9, 13-14 and 16-17 yr. The Met allele was associated with symptoms of ADHD at ages 8-9 and 13-14 yr. A multivariate regression analysis revealed that the observed effect of the Met allele on ADHD symptoms reflects an influence on persistent hyperactivity-impulsivity symptoms. The present findings support the hypothesis that BDNF is involved in the pathogenesis of ADHD. The results highlight the importance of distinguishing between hyperactivity-impulsivity and inattention, respectively, and demonstrate the value of using a longitudinal approach in genetic studies of ADHD symptoms.


In vivo profile of ICA-27243 [N-(6-chloro-pyridin-3-yl)-3,4-difluoro-benzamide], a potent and selective KCNQ2/Q3 (Kv7.2/Kv7.3) activator in rodent anticonvulsant models.
Openers or activators of neuronal KCNQ2/Q3 potassium channels decrease neuronal excitability and may provide benefit in the treatment of disorders of neuronal excitability such as epilepsy. In the present study, we evaluate the effects of ICA-27243 [N-(6-chloro-pyridin-3-yl)-3,4-difluoro-benzamide], an orally bioavailable, potent, and selective KCNQ2/Q3 opener, in a broad range of rodent seizure models. ICA-27243 was effective against maximal electroshock (MES) and pentylenetetrazole (PTZ)-induced seizures in both rats (MES, ED(50) = 1.5 mg/kg p.o.; PTZ, ED(50) = 2.2 mg/kg p.o.) and mice (MES, ED(50) = 8.6 mg/kg p.o.; PTZ, ED(50) = 3.9 mg/kg p.o.) in the rat amygdala kindling model of partial seizures (full protection from seizure at 9 mg/kg p.o.) and in the 6-Hz model of psychomotor seizures in mice (active at 10 mg/kg i.p.). Antiseizure efficacy in all models was observed at doses significantly less than those shown to effect open-field locomotor activity (rat ED(50) = 40 mg/kg p.o.) or ability to remain on a Rotorod (no effect in rat at doses up to 100 mg/kg p.o.). There was no evidence of cognition impairment as measured in the Morris water maze in the rat (10 and 30 mg/kg p.o.), nor was there evidence of the development of tolerance after multiple doses of ICA-27243. Our findings suggest that selective KCNQ2/Q3 opening activity in the absence of effects on KCNQ3/Q5 or GABA-activated channels may be sufficient for broad-spectrum antiepileptic activity in rodents.


System-agnostic clinical decision support services: benefits and challenges for scalable decision support.
System-agnostic clinical decision support (CDS) services provide patient evaluation capabilities that are independent of specific CDS systems and system implementation contexts. While such system-agnostic CDS services hold great potential for facilitating the widespread implementation of CDS systems, little has been described regarding the benefits and challenges of their use. In this manuscript, the authors address this need by describing potential benefits and challenges of using a system-agnostic CDS service. This analysis is based on the authors' formal assessments of, and practical experiences with, various approaches to developing, implementing, and maintaining CDS capabilities. In particular, the analysis draws on the authors' experience developing and leveraging a system-agnostic CDS Web service known as SEBASTIAN. A primary potential benefit of using a system-agnostic CDS service is the relative ease and flexibility with which the service can be leveraged to implement CDS capabilities across applications and care settings. Other important potential benefits include facilitation of centralized knowledge management and knowledge sharing; the potential to support multiple underlying knowledge representations and knowledge resources through a common service interface; improved simplicity and componentization; easier testing and validation; and the enabling of distributed CDS system development. Conversely, important potential challenges include the increased effort required to develop knowledge resources capable of being used in many contexts and the critical need to standardize the service interface. Despite these challenges, our experiences to date indicate that the benefits of using a system-agnostic CDS service generally outweigh the challenges of using this approach to implementing and maintaining CDS systems.


Measuring and decomposing socioeconomic inequality in healthcare delivery: A microsimulation approach with application to the Palestinian conflict-affected fragile setting.
Socioeconomic-related inequalities in healthcare delivery have been extensively studied in developed countries, using standard linear models of decomposition. This paper seeks to assess equity in healthcare delivery in the particular context of the occupied Palestinian territory: the West Bank and the Gaza Strip, using a new method of decomposition based on microsimulations. Besides avoiding the 'unavoidable price' of linearity restriction that is imposed by the standard methods of decomposition, the microsimulation-based decomposition enables to circumvent the potentially contentious role of heterogeneity in behaviours and to better disentangle the various sources driving inequality in healthcare utilisation. Results suggest that the worse-off do have a disproportinately greater need for all levels of care. However with the exception of primary-level, utilisation of all levels of care appears to be significantly higher for the better-off. The microsimulation method has made it possible to identify the contributions of factors driving such pro-rich patterns. While much of the inequality in utilisation appears to be caused by the prevailing socioeconomic inequalities, detailed analysis attributes a non-trivial part (circa 30% of inequalities) to heterogeneity in healthcare-seeking behaviours across socioeconomic groups of the population. Several policy recommendations for improving equity in healthcare delivery in the occupied Palestinian territory are proposed.


Amplified spontaneous emission in thin films of quasi-2D BA3MA3Pb5Br16 lead halide perovskites.
Quasi-2D (two-dimensional) hybrid perovskites are emerging as a new class of materials with high photoluminescence yield and improved stability compared to their three-dimensional (3D) counterparts. Nevertheless, despite their outstanding emission properties, few studies have been reported on amplified spontaneous emission (ASE) and a thorough understanding of the photophysics of these layered materials is still lacking. In this work, we investigate the ASE properties of multilayered quasi-2D BA3MA3Pb5Br16 films through the dependence of the photoluminescence on temperature and provide a novel insight into the emission processes of quasi-2D lead bromide perovskites. We demonstrate that the PL and ASE properties are strongly affected by the presence, above 190 K, of a minor fraction of the high temperature (HT) phase. This phase dominates the PL spectra at low excitation density and strongly affects the ASE properties. In particular, ASE is only present between 13 K and 230 K, and, at higher temperatures, it is suppressed by absorption of charge transfer states of the HT phase. Our results improve the understanding of the difficulties to obtain ASE at room temperature from these quasi-2D materials and are expected to guide possible materials improvement in order to exploit their excellent emission properties also for the realization of low threshold optically pumped lasers.


Increased Frequency of Heterozygous Alpha-1-Antitrypsin Deficiency in Liver Explants From Nonalcoholic Steatohepatitis Patients.
Cirrhotic explanted livers occasionally have unexpected periodic acid-Schiff-diastase (PASD)-positive globules within the hepatocyte cytoplasm. It is often unclear whether this finding is a nonspecific consequence of cirrhosis or is indicative of an underlying alpha-1-antitrypsin deficiency (A1ATD) contributing to the cirrhosis. In this study, explanted livers were retrospectively evaluated for histopathology (including PASD status with confirmatory alpha-1-antitrypsin [A1AT] immunohistochemistry [IHC]), and chart review provided etiology of liver failure and general clinical parameters. Real-time polymerase chain reaction was used to detect A1AT genotype (SERPINA1 S and Z alleles) by melting curve analysis on liver explant tissue from selected cases. Of 196 explanted livers, 21 (11%) had PASD+ globules, which were significantly enriched in patients with a clinical diagnosis of nonalcoholic steatohepatitis (NASH; 47%) compared with other causes (P < 0.001). IHC confirmed all PASD+ globules were A1AT+, with 20 of 21 cases demonstrating diffuse A1AT staining. In an expanded NASH cohort, 42% (14/33) of explants had PASD+ globules, 92% of which were homozygous (n = 1) or heterozygous (n = 11) for the SERPINA1 Z allele, corresponding to nearly 40% of all NASH patients. Overall, the Z allele was present in 10% of all tested liver explants, with 85% of PASD+ cases genotyping homozygous (n = 2) or heterozygous (n = 20), which is far in excess of the estimated 2% in the general population. These results indicate PASD+ A1AT globules (with confirmatory genotyping showing at least 1 Z allele) are commonly observed in NASH, suggesting a synergistic relationship toward liver fibrosis. In addition, the high frequency of SERPINA1 Z alleles in liver transplantation patients supports the utility of pretransplant genotyping.


Evaluating the implementation of a person-centred transition programme for adolescents and young adults with long-term conditions: the role of context and organisational behaviour.
Drawing on the experiences of healthcare professionals in one paediatric hospital, this paper explores the influence of context and organisational behaviour on the implementation of a person-centred transition programme for adolescents and young adults (AYA) with long-term conditions. A single embedded qualitative case study design informed by a realist evaluation framework, was used. Participants who had experience of implementing the transition programme were recruited from across seven individual services within the healthcare organisation. The data were gathered through semi-structured interviews (n = 20) and analysed using thematic analysis. Implementation of the transition programme was influenced by the complex interaction of macro, meso and micro processes and contexts. Features of organisational behaviour including routines and habits, culture, organisational readiness for change and professional relationships shaped professional decision-making around programme implementation. There exists a significant body of research relating to the role of context and its influence on the successful implementation of complex healthcare interventions. However, within the area of healthcare transition there is little published evidence on the role that organisational behaviour and contextual factors play in influencing transition programme implementation. This paper provides an in-depth understanding of how organisational behaviour and contextual factors affect transition programme implementation.


An exploratory study of BDNF and oxidative stress marker alterations in subacute and chronic stroke patients affected by neuropathic pain.
Patients affected by stroke, particularly subacute stroke patients, often complain of neuropathic pain (NP), which may severely impair their quality of life. The aim of this exploratory study was to characterize NP and to investigate whether there is a correlation between NP and serum brain-derived neurotrophic factor (BDNF) and serum markers of oxidative stress. We enrolled 50 patients divided in subacute (< 90 days from stroke onset) and chronic (> 90 and 180 < days from stroke onset). The Barthel Index, Deambulation Index, and Short Form 36 were used to assess the patients' degree of disability and quality of life. Pain-specific tools, namely the Numeric Rating Scale (NRS), Neuropathic Pain Diagnostic questionnaire (DN4), and Neuropathic Pain Symptom Inventory (NPSI), were also used. Serum levels of BDNF and markers of oxidative stress and of metal status were evaluated: copper, iron, transferrin, ferritin, ceruloplasmin concentration (iCp) and activity (eCp), Total Antioxidant status (TAS), Cp/Tf ratio, eCp/iCp ratio, and non-ceruloplasmin bound copper (Non-Cp Cu). We found the highest value of BDNF in subacute with NP (DN4 score ≥ 4). The TAS, Cp/Tf ratio, and eCp/iCp not only fell outside the normal reference range in a high percentage of subacute and chronic patients, but were also found to be related to specific NP symptoms. These preliminary results reveal altered BDNF and oxidative stress indices in subacute stroke patients who complain of NP. These investigative findings may shed more light on the mechanisms underlying NP and consequently lead to a more tailored therapeutic and/or rehabilitation procedure of subacute stroke patients.


Pole-vaulting: identification of the pole local bending rigidities by an updating technique.
The development of composite material poles since 1960 has played a prominent part in performance improvement in pole-vaulting. Previous studies devoted to pole-vaulting models were based on constant mechanical characteristics. It is thus necessary to identify the local bending rigidities of the pole to build realistic pole-vaulting models. Updating methods developed for dynamic structure studies allow us to describe local mechanical characteristics. These methods are based on the comparison between experimental results and those determined numerically by finite element models. This study presents an adaptation of these methods to determine the local bending rigidities of the pole. The updating technique is validated by a deflection test of a homogeneous beam. Then, a study of the model sensitivity is carried out to investigate the procedure robustness. Finally, the updating method is applied to an old design pole and to a recent one. The results obtained vary greatly from one pole to the other; they highlight the evolutions in pole design.


Post-transcriptional expression regulation in the yeast Saccharomyces cerevisiae on a genomic scale.
Based on large-scale data for the yeast Saccharomyces cerevisiae (protein and mRNA abundance, translational status, transcript length), we investigate the relation of transcription, translation, and protein turnover on a genome-wide scale. We elucidate variations between different spatial cell compartments and functional modules by comparing protein-to-mRNA ratios, translational activity, and a novel descriptor for protein-specific degradation (protein half-life descriptor). This analysis helps to understand the cell's strategy to use transcriptional and post-transcriptional regulation mechanisms for managing protein levels. For instance, it is possible to identify modules that are subject to suppressed translation under normal conditions ("translation on demand"). In order to reduce inconsistencies between the datasets, we compiled a new reference mRNA abundance dataset and we present a novel approach to correct large microarray signals for a saturation bias. Accounting for ribosome density based on transcript length rather than ORF length improves the correlation of observed protein levels to translational activity. We discuss potential causes for the deviations of these correlations. Finally, we introduce a quantitative descriptor for protein degradation (protein half-life descriptor) and compare it to measured half-lives. The study demonstrates significant post-transcriptional control of protein levels for a number of different compartments and functional modules, which is missed when exclusively focusing on transcript levels.


Don't Joke About Me: Student Identities and Perceptions of Instructor Humor in College Science Courses.
Humor is a popular tool used by instructors to engage students. However, some instructor jokes may be perceived as less funny and more offensive by particular groups of students. Previous studies have shown that student gender impacts student perception of instructor humor; however, to our knowledge no studies have explored whether there are differences in how other identity groups interpret instructor humor. In this study, we surveyed 1,637 students across 25 different college science courses at a research-intensive institution in the Southwest United States. Students evaluated a set of topics that science instructors might joke about in class as to whether they were funny and offensive. Using binary logistic regression, we analyzed whether students of different identities, including race/ethnicity, political affiliation, LGBTQ+ status, religious affiliation, and native language, differentially perceived joke topics to be funny and offensive if told by an instructor in class. We identified that topics which tended to be perceived by students as funny rather than offensive were generally less likely to be perceived as funny to non-native English language speakers compared with native English speakers. We also found that students were more likely to be offended by jokes about their own identity group. This work identifies potentially humorous topics that instructors should avoid because they could be offensive to groups of students. This study also highlights topics that tend to be perceived as funny to most students, which indicates that instructors who joke about such topics may be universally benefitting college science students.


[Genetic polymorphism in three indigenous ethnic populations of Dagestan].
Genetic, demographic, and biochemical studies of three isolated ethnic populations of Dagestan were performed. Two populations lived in their historical (highland and lowland) habitats, and one population migrated from highlands to lowlands, i.e., changed its environment. A substantial genetic differentiation with respect to eight biochemical systems and marital structure was found. Altitudinal variation was revealed in allelic frequencies of group-specific component (GC) and haptoglobin (HP). The frequency of allele GC * 1 decreased and that of allele HP * 1 increased as the altitude increased in the studied localities. The studied populations differed with respect to the average heterozygosity levels per locus. Both absolute heterozygosity levels and the pattern of interpopulation differences estimated for all of the eight biochemical markers considerably differed from those estimated for all markers except for GC and HP. These data indicate that the latter two biochemical systems are important for adaptation of the studied Dagestan populations. In surviving migrants from highlands to lowlands, the levels of inbreeding and genetic polymorphism were intermediate between these values for highlanders and native lowlanders. Earlier, the 30% selective mortality was revealed in migrants from highlands to lowlands during the first years of adaptation to new conditions. Those individuals who died had a higher inbreeding level, homozygosity, and physiological sensitivity. In this study, only surviving migrants were examined. Therefore, it may be suggested that the inadaptive (dying) part of migrants also differed from the adaptive (surviving) part in the higher HP * 1 and GC * 2 allelic frequencies.


Production, characterization, and reconstitution of recombinant quinoprotein glucose dehydrogenase (soluble type; EC 22.214.171.124) apoenzyme of Acinetobacter calcoaceticus.
Soluble, periplasmic quinoprotein glucose dehydrogenase of Acinetobacter calcoaceticus (sGDH; EC 22.214.171.124) was produced in good yield in the apoenzyme form (without the cofactor pyrroloquinoline quinone, PQQ) by an Escherichia coli recombinant strain provided with a plasmid containing the gene under control of a lac promoter. Structural analysis of the purified apoenzyme revealed that the E. coli strain used produces the correct mature protein. Titration of the apoenzyme with PQQ in the presence of Ca2+ showed that a linear relation exists between the amount of added PQQ and activity observed, and that the subunit and PQQ associate in a molar ratio of 1:1. Based on spectral and enzymatic criteria, it is concluded that the present holoenzyme preparation has a better quality than the previously described preparations of authentic holoenzyme. As isolated here, the recombinant apoenzyme was in the dimeric form. Partial monomerization occurred upon gel filtration in a buffer with chelator and the process could be reversed with Ca2+. PQQ binds to the dimer in the presence of chelator, not to the monomer. However, the PQQ-containing dimer was not active and showed an unusual absorption spectrum which was slowly converted into a PQQH2-like spectrum when glucose was added. Full restoration of activity was achieved upon addition of Ca2+ and the spectra were immediately converted into those of normal holoenzyme in the oxidized and reduced form, respectively. Addition of chelator to holoenzyme did not lead to inactivation or monomerization. It is concluded, therefore, that Ca2+ has a dual role in this enzyme, being required for dimerization of the subunits as well as for functionalization of the bound PQQ, and that it is more firmly attached to the holoenzyme than to the apoenzyme.


Distribution in rat brain of binding sites of kaliotoxin, a blocker of Kv1.1 and Kv1.3 alpha-subunits.
The distribution of the binding sites for kaliotoxin (KTX), a blocker of voltage-dependent K(+) channels, was studied with quantitative autoradiography in adult rat brain and during postnatal brain maturation. Iodinated KTX bound specifically to tissue sections with a high affinity (K(d) = 82 pM) and a maximal binding capacity of 13.4 fmol/mg protein. The distribution of KTX binding sites within the central nervous system was heterogeneous. The highest densities were found in the neocortex, hypothalamus, dentate gyrus, bed nucleus of the stria terminalis, and parabrachial nuclei. The lowest level was observed in the white matter. From postnatal day 5 onward, KTX binding sites were detectable only in the hindbrain. The density of KTX binding sites in whole brain drastically increased after postnatal day 15 to achieve adult levels at postnatal day 60 in the whole brain. Bath application of KTX to Xenopus laevis oocytes blocked recombinant Kv1.3 and Kv1.1 channels potently and Kv1.2 channels less potently, with respective K(d) values of 0.1, 1.5, and 25 nM. KTX affinities for each of these channels expressed in mammalian cells were about 10-fold lower. A comparison of the distribution of KTX binding sites with that of Kv1 channel polypeptides, together with the pharmacology of KTX block, suggests that the principal targets for KTX in rat brain are K(+) channels containing Kv1.1 and Kv1.3 alpha-subunits.


Contemporary Patients Have Better Perioperative Outcomes Following Cytoreductive Nephrectomy: A Multi-institutional Analysis of 1272 Consecutive Patients.
To evaluate factors associated with perioperative outcomes in a multi-institutional cohort of patients treated with cytoreductive nephrectomy (CN). Data were analyzed for metastatic renal cell carcinoma patients treated with CN at 6 tertiary academic centers from 2005 to 2019. Outcomes included: Clavien-Dindo complications, mortality, length of hospitalization, 30-day readmission rate, and time to systemic therapy. Univariate and multivariable models evaluated associations between outcomes and prognostic variables including the year of surgery. A total of 1272 consecutive patients were treated with CN. Patients treated in 2015-2019 vs 2005-2009 had better performance status (P<.001), higher pathologic N stage (P = .04), more frequent lymph node dissections (P<.001), and less frequent presurgical therapy (P = .02). Patients treated in 2015-2019 vs 2005-2009 had lower overall and major complications from surgery, 22% vs 39%, P<.001% and 10% vs 16%, P = .03. Mortality at 90days was higher for patients treated 2005-2009 vs 2015-2019; 10% vs 5%, P = .02. After multivariable analysis, surgical time period was an independent predictor of major complications and 90-day mortality following cytoreductive surgery. Postoperative major complications and mortality rates following CN are significantly lower in patients treated within the most recent time period.


Association of Atrial Fibrillation Without Cardiovascular Comorbidities and Stroke Risk: From the REGARDS Study.
Background Atrial fibrillation (AF) is associated with a 5-fold increased stroke risk. While most patients with AF warrant anticoagulation, optimal treatment remains uncertain for patients with AF without cardiovascular comorbidities because the risk of stroke in this population has not been well-characterized. Methods and Results Participants (N=28 253; 55% women, mean age 64.6±9.4 years), from the REGARDS (Reasons for Geographic and Racial Differences in Stroke) study (2003-present) were classified into 1 of 4 groups based on the presence or absence of AF and the presence or absence of cardiovascular comorbidities. Cox proportional hazards analysis was used to compare the risk of stroke between groups. During 244 560 person-years of follow-up (median 8.7 years), 1206 strokes occurred. Compared with patients with neither AF nor cardiovascular comorbidities, we did not find an increased stroke risk (hazard ratio [HR], 1.23; 95% CI, 0.62-2.18 [P=0.511]) among participants with AF alone. Participants without AF but with cardiovascular comorbidities had both an elevated stroke risk (HR, 1.77; 95% CI, 1.48-2.18 [P<0.0001]) and an increased risk of cardioembolic stroke (HR, 2.34; 95% CI, 1.48-3.90 [P=0.0002]). Conclusions In this large cohort of participants with AF without cardiovascular comorbidities, we found that AF itself, without cardiovascular comorbidities, did not confer increased risk of stroke. Cardiovascular comorbidities, however, were associated with an increased risk of both stroke of any type and cardioembolic stroke, even in the absence of AF.


Sex differences in function and distribution of alpha 1- and alpha 2-adrenoceptors in rabbit urethra.
To define the functional role of postsynaptic alpha-adrenoceptors in urethral smooth muscle, we examined the effects of various alpha-adrenergic agonists and antagonists on the contractile properties of the isolated urethra of male and female rabbits and quantified the population of alpha 1-and alpha 2-adrenoceptors, using radioligand receptor binding techniques. Norepinephrine (NE), epinephrine, and phenylephrine, an alpha 1-adrenergic agonist, induced increases in contractile force in both the male and female urethra. Clonidine, an alpha 2-adrenergic agonist, caused a relatively large contractile response in the female urethra but only a small contractile response in the male urethra. Receptor binding studies indicated that the male urethra contains almost equal amounts of alpha 1- and alpha 2-adrenoceptors (32 vs. 34 fmol/mg, respectively), whereas the female urethra contains a significantly greater density of alpha 2- than alpha 1-adrenoceptors (122 vs. 34 fmol/mg, respectively). Our studies indicate that both alpha 1- and alpha 2-adrenoceptors cause contractile responses in male and female rabbit urethra; and the greater response to alpha 2-agonist in female than male urethra is correlated with a higher density of alpha 2-adrenoceptors in this tissue.


Medium-term scenarios of COVID-19 as a function of immune uncertainties and chronic disease.
As the SARS-CoV-2 trajectory continues, the longer-term immuno-epidemiology of COVID-19, the dynamics of Long COVID, and the impact of escape variants are important outstanding questions. We examine these remaining uncertainties with a simple modelling framework that accounts for multiple (antigenic) exposures via infection or vaccination. If immunity (to infection or Long COVID) accumulates rapidly with the valency of exposure, we find that infection levels and the burden of Long COVID are markedly reduced in the medium term. More pessimistic assumptions on host adaptive immune responses illustrate that the longer-term burden of COVID-19 may be elevated for years to come. However, we also find that these outcomes could be mitigated by the eventual introduction of a vaccine eliciting robust (i.e. durable, transmission-blocking and/or 'evolution-proof') immunity. Overall, our work stresses the wide range of future scenarios that still remain, the importance of collecting real-world epidemiological data to identify likely outcomes, and the crucial need for the development of a highly effective transmission-blocking, durable and broadly protective vaccine.