Adaptation of bacterial community in maize rhizosphere for enhancing dissipation of phthalic acid esters in agricultural soil.
Rhizospheric degradation is a green and in situ strategy to accelerate dissipation of organic pollutants in soils. However, the mechanism on microbial degradation of phthalic acid esters (PAEs) in rhizosphere is still unclear. Here, the bacterial community and function genes in bulk and rhizospheric soils of maize (Zea mays L.) exposed to gradient concentrations of di-(2-ethylhexyl) phthalate (DEHP) were analyzed with 16 S rRNA, metagenomic sequencing and quantitative PCR (qPCR). Maize rhizosphere significantly increased the dissipation of DEHP by 4.02-11.5% in comparison with bulk soils. Bacterial community in rhizosphere exhibited more intensive response and shaped its beneficial structure and functions to DEHP stress than that in bulk soils. Both rhizospheric and pollution effects enriched more PAE-degrading bacteria (e.g., Bacillus and Rhizobium) and function genes in rhizosphere than in bulk soil, which played important roles in degradation of PAEs in rhizosphere. The PAE-degrading bacteria (including genera Sphingomonas, Sphingopyxis and Lysobacter) identified as keystone species participated in DEHP biodegradation. Identification of PAE intermediates and metagenomic reconstruction of PAE degradation pathways demonstrated that PAE-degrading bacteria degraded PAEs through cooperation with PAE-degrading and non-PAE-degrading bacteria. This study provides a comprehensive knowledge for the microbial mechanism on the superior dissipation of PAEs in rhizosphere.


Effects of fluid flow shear rate and surface roughness on the calcification of polymeric heart valve leaflet.
Surface defects, blood flow shear rates and mechanical stresses are contributing factors in the calcification process of polymeric devices exposed to the blood flow. A number of experiments were performed to evaluate the effect of surface defects such as roughness and cracks and flow shear rate on the calcification process of a polyurethane material used in the design of prosthetic heart valves. Results showed that polyurethane surface gets calcified and the calcification is more pronounced at the lower shear rates. Roughness and cracks both increase the calcification levels. The results also suggest very little diffusion of calcium to the subsurface indicating that calcification of a polyurethane material, is a surface phenomenon. Based on a simple peeling test, the bond strength between the calcified layer and polyurethane was found to be extremely weak, suggesting that the bonding is in the form of Van-der-Waals. A limited set of experiments with polycarbonate showed that polycarbonate is less prone to calcification compared to polyurethane (p values less than 0.05), indicating its potential application in medical devices exposed to blood flow.


RGB-D fusion models for construction and demolition waste detection.
The development of urbanization has brought a large amount of construction and demolition waste (CDW), which occupy land and cause adverse ecological effects. To effectively solve the negative impact of CDW, it needs to be recycled. Accurate waste classification is key to successful waste management. However, the current waste classification methods mainly use color images to classify, which cannot meet the needs of accurate classification. This paper built an RGB-depth (RGB-D) detection platform, using a color camera and a laser line-scanning sensor to collect RGB images and depth images. In order to use RGB images and depth images for feature fusion more effectively, this paper proposed three fusion models: RGB-D concat、RGB-D Ci-add and RGB-D Ci-concat. All these models based on an instance segmentation network called mask region convolutional neural network (Mask R-CNN), which can accurately segment the contours of each object while classifying them. The experimental results show that the mAPs of the RGB-D Ci-add / concat model are 1.33% to 1.72% higher than those of the RGB model, and the classification accuracy is 1.92% ∼ 2.27% higher. In addition, all the proposed models can meet the real-time requirement of online detection. Due to the excellent comprehensive performance of the RGB-D Ci-concat model, it can be regarded as the final detection model of the robot, which can improve the sorting efficiency of CDW further.


The effectiveness and safety of beta antagonist in burned patients: A systematic review and meta-analysis.
Beta antagonist is one of the most effective and the least toxic pharmacological treatments to attenuate the raised catecholamine effects for burned patients. To evaluate the effectiveness and safety of beta blocker compared with placebo or usual care in burned patients, a meta-analysis of randomised controlled trials (RCTs) was conducted. We searched the database of PubMed, Embase, the Cochrane Library, and Web of Science to 10 April 2020. Two investigators independently assessed articles for inclusion and exclusion criteria and selected studies for the final analysis. We performed the meta-analysis using a random-effect model. A total of 12 RCTs were included in the study, including 1887 patients. Propranolol-treated patients have a decrease in length of hospital stay in adults (weighted mean difference [WMD] = -9.06, 95% CIs = [-12.88, -5.24]) and prepare time of graft (WMD = -7.88, 95% CIs = [-12.27, -3.50]). Similarly, the use of propranolol could significantly decrease heart rate (WMD = -15.16, 95% CIs = [-20.37, -9.94]), rate pressure product (WMD = -1.32, 95% CIs = [-1.67, -0.97]), and mean arterial pressure (WMD = -2.75, 95% CIs = [-4.23, -1.26]). Moreover, there is no significant difference between propranolol and placebo with respect to mortality (risk difference [RD] = 0.00, 95% CIs [-0.03, 0.04]), sepsis (RD = -0.03, 95% CIs [-0.09, 0.03]), and events of post-traumatic stress disorder (PTSD) and acute stress disorder (RD = -0.01, 95% CIs [-0.07, 0.05]), and also, there is no significant difference in subgroup analysis based on age. The use of beta antagonist in burned patients does reduce length of hospital stay in adults, shorten the preparation time for graft, and reduce heart burden, without increasing mortality, sepsis, or PTSD compared with those who had usual care or placebo. So beta antagonist can be considered as an appropriate treatment strategy in burned patients. More prospective, randomised-controlled, multi-centre studies were needed to define their place in therapeutic algorithms.


Three-screw versus two-screw fixation of distal fragment in fifth metacarpal neck fractures stabilized with locking plate.
Fifth metacarpal neck fracture commonly requires open reduction and internal fixation. Locking plate was widely adopted in the treatment of fifth metacarpal neck fracture as first-line choice for fixation. Patients with fifth metacarpal neck fracture receiving locking plate fixation were included for analysis. Features of internal fixation including number of distal and proximal locking screws, diameter of the screws and usage of lag screws were recorded. Clinical and radiographic outcomes included final volar angulation, grip strength, Michigan Hand Outcomes Questionnaire (MHQ) and range of motion (ROM) of fifth metacarpophalangeal joint. Three-screw fixation was less frequently presented in the group with increased volar angulation (≥30 degrees). Consistently, three-screw fixation of distal fragment could improve the prognosis compared with two-screw fixation (MHQ 95.4 ± 5.1 versus 80.4 ± 12.3, ROM 83.5 ± 7.2 versus 69.6 ± 7.7). In conclusion, the metacarpal head should be fixed by three locking screws instead of two locking screws.


Dissemination and implementation of prolonged exposure therapy for posttraumatic stress disorder.
Posttraumatic stress disorder (PTSD) is a highly prevalent, often chronic and disabling psychiatric disorder that is associated with significant adverse health and life consequences. Although several evidence-based treatments (EBTs), including Prolonged Exposure therapy (PE), have been found effective and efficacious in reducing PTSD symptomology, the majority of individuals with this disorder receive treatments of unknown efficacy. Thus, it is imperative that EBTs such as PE be made available to PTSD sufferers through widespread dissemination and implementation. We will review some of the efforts to increase the availability of PE and the common barriers to successful dissemination and implementation. We also discuss novel dissemination strategies that are harnessing technology to overcome barriers to dissemination.


Recognizing the intensity of strength training exercises with wearable sensors.
In this paper we propose a system based on a network of wearable accelerometers and an off-the-shelf smartphone to recognize the intensity of stationary activities, such as strength training exercises. The system uses a hierarchical algorithm, consisting of two layers of Support Vector Machines (SVMs), to first recognize the type of exercise being performed, followed by recognition of exercise intensity. The first layer uses a single SVM to recognize the type of the performed exercise. Based on the recognized type a corresponding intensity prediction SVM is selected on the second layer, specializing in intensity prediction for the recognized type of exercise. We evaluate the system for a set of upper-body exercises using different weight loads. Additionally, we compare the most important features for exercise and intensity recognition tasks and investigate how different sliding window combinations, sensor configurations and number of training subjects impact the algorithm performance. We perform all of the experiments for two different types of features to evaluate the feasibility of implementation on resource constrained hardware. The results show the algorithm is able to recognize exercise types with approximately 85% accuracy and 6% intensity prediction error. Furthermore, due to similar performance using different types of features, the algorithm offers potential for implementation on resource constrained hardware.


Clinical analysis of 156 cases of multiple organ failure caused by fish bile.
To analyze the clinical features and prognosis of patients suffering from fish bile poisoning. A total of 156 multiple organ failure (MOF) patients caused by fish bile poisoning were hospitalized in our department over the past 28 years. The patients' symptoms, examination results, treatment and outcomes were collected and analyzed. All patients' first symptom was gastrointestinal discomfort, including unbearable nausea and intractable vomiting. The symptoms that followed were oliguria or anuria, edema, headache, fatigue, jaundice, palpitation, alimentary tract hemorrhage, gross hematuria, dyspnea, shock, tachycardia, bradycardia, arrhythmia, coma, and cardiac arrest. The symptom severity and cohort were different among different patients. Twenty-one cases received gastroscopy, which exhibited diffuse gastric mucosal bleeding. Twelve patients received renal biopsy, which exhibited focal necrosis of tubular epithelial cells. One patient received a liver biopsy, which exhibited extensive hepatocyte necrosis. All patients received blood purification therapy. Of the four patients who died, 4 out of 5 organs had failed. The general mortality rate was 2.6%. Compared with the MOF caused by trauma and sepsis, the fish bile poisoning MOF has a much lower morality rate. However, patients with higher age, more underlying diseases, and more organ failure tended to have a worse prognosis.


Modelling the extra and intracellular uptake and discharge of heavy metals in Fontinalis antipyretica transplanted along a heavy metal and pH contamination gradient.
Samples of the aquatic bryophyte Fontinalis antipyretica Hedw. were transplanted to different sites with the aim of characterizing the kinetics of the uptake and discharge of heavy metals in the extra and intracellular compartments. The accumulation of metals in extracellular compartments, characterized by an initial rapid accumulation, then a gradual slowing down over time, fitted perfectly to a Michaelis-Menten model. The discharge of metals from the same compartment followed an inverse linear model or an inverse Michaelis-Menten model, depending on the metal. In intracellular sites both uptake and discharge occurred more slowly and progressively, following a linear model. We also observed that the acidity of the environment greatly affected metal accumulation in extracellular sites, even when the metals were present at relatively high concentrations, whereas the uptake of metals within cells was much less affected by pH.


The haematogenous spread of tumor: a prevental process.
The establishment of a secondary tumor is the outcome of a series of biological events. Cells detach from the primary tumor and penetrate vascular channels, which they leave again at a distant site to implant and proliferate in an organ or tissue remote from the primary tumor. These processes depend on the adhesion of tumor cells to each other, on the permeability of the venous or lymphatic channels and on the susceptibility of the substrate. Present experiments show that these properties are modifiable and that haematogenous tumor dissemination is a preventable process.


Genotoxicity and inactivation of catechol metabolites of the mycotoxin zearalenone.
Zearalenone (ZEN) is a highly estrogenic mycotoxin produced by Fusarium species. The adverse effects of ZEN and its reductive metabolite α-zearalenol (α-ZEL) are often compared to those of 17β-estradiol (E2) and estrone (E1). These endogenous steroidal estrogens are associated with an increased risk for cancer, which may be mediated by two mechanisms, i.e. (1) hormonal activity and (2) genotoxic effects after cytochrome P450-catalyzed metabolic activation to catechols. Like E1 and E2, ZEN and α-ZEL exhibit marked estrogenicity and also undergo aromatic hydroxylation to catechol metabolites. The subsequent methylation of catechols by catechol-O-methyltransferase (COMT) is generally considered as a detoxifying pathway. Imbalances between the activation and inactivation reactions can lead to the formation of reactive semiquinones and quinones, which can alkylate DNA or produce reactive oxygen species by redox cycling. In the present study, the genotoxicity of the catechol metabolites of ZEN, α-ZEL, E1 and E2 was determined in a cell-free system by measuring 8-oxo-2'-deoxyguanosine using a LC-DAD-MS(2) method. Each of the individual catechols of ZEN, α-ZEL, E1 and E2 induced oxidative DNA damage in calf thymus DNA. The ranking order of the DNA damaging activity was 15-hydroxy-ZEN/α-ZEL ≈ 2/4-hydroxy-E1/E2 > 13-hydroxy-ZEN/α-ZEL. When hepatic microsomes from different species were incubated with ZEN, the rat had the highest activity for catechol formation, followed by human, mouse, pig and steer. The amount of catechol metabolites correlated directly with the amount of oxidative damage in calf thymus DNA. The ranking order for the rate of methylation by human hepatic COMT was 2-hydroxy-E1/E2 >> 4-hydroxy-E1/E2 >> 13/15-hydroxy-ZEN/α-ZEL. Thus, the catechol metabolites of the mycoestrogen ZEN and its reductive metabolite α-ZEL exhibit a DNA-damaging potential comparable to that of the catechol metabolites of E1 and E2, but are much poorer substrates for inactivation by human COMT.


Characterization of a thermostable Bacillus stearothermophilus alpha-amylase.
Liquefying-type Bacillus stearothermophilus alpha-amylase was characterized. The coding gene was cloned in Bacillus subtilis and the enzyme was produced in three different host organisms: B. stearothermophilus, B. subtilis, and Escherichia coli. Properties of the purified enzyme were similar irrespective of the host. Temperature optimum was at 70-80 degrees C and pH optimum at 5.0-6.0. The enzyme was stable for 1 h in the pH range 6.0-7.5 at 80 degrees C. The enzyme was stabilized by Ca2+, Na+, and bovine serum albumin. About 50% of the activity remained after heating at 70 degrees C for 5 days or 45 min at 90 degrees C. Metal ions Cd2+, Cu2+, Hg2+, Pb2+, and Zn2+ were inhibitory, whereas EDTA, ethylene glycol bis(beta-aminoethyl ether) N,N,N',N'-tetraacetic acid, and Tendamistat were without effect. The enzyme was fully active after treatment in acetone or ethanol at 55 or 70 degrees C, respectively, for 30 min. Sodium dodecyl sulfate (1%) did not affect stability, whereas 6 M urea denatured totally at 70 degrees C. The Km value for soluble starch was 14 mg/ml. Mr is 59,000 and pI 8.8. The only difference between the enzymes produced in different hosts was in signal peptide processing.


Preferences of patients, clinicians, and healthy controls for the management of a Bethesda III thyroid nodule.
Active surveillance is propagated as an alternative for hemithyroidectomy in the management of Bethesda III thyroid nodules. A cross-sectional survey questioned respondents on their willingness to accept risks related to active surveillance and hemithyroidectomy. In case of active surveillance, respondents (129 patients, 46 clinicians, and 66 healthy controls) were willing to accept a risk of 10%-15% for thyroid cancer and 15% for needing more extensive surgery in the future. Respondents were willing to accept a risk of 22.5%-30% for hypothyroidism after hemithyroidectomy. Patients and controls were willing to accept a higher risk on permanent voice changes compared with clinicians (10% vs. 3%, p < 0.001). Real-life risks associated which active surveillance and hemithyroidectomy for Bethesda III nodules are equivalent or less than the risks people are willing to accept. Clinicians accepted less risk for permanent voice changes.


[What is the risk of Takotsubo in women?]
Takotsubo cardiomyopathy preferentially affects postmenopausal women (# 90%). Takotsubo cardiomyopathy mimics an acute coronary syndrome and is defined as a transient left ventricular systolic dysfunction. Diagnosis is based on the criteria of the Mayo Clinic. Stress is found in around 80% of cases: physical stress, emotional stress, medical stress. Physical stress is less found in women. Incidence of Takotsubo cardiomyopathy is 54.9/1,000,000 inhabitants in women versus only 3.6/1,000,000 inhabitants in men. Diagnosis is based on electrocardiogram, biomarkers, echocardiography, coronary angiography and left ventricular angiography, and cardiac magnetic imaging. The complete recovery of left ventricular systolic function must be assessed, allowing to confirm the diagnosis of Takotsubo cardiomyopathy.


Ion selectivity in potassium channels.
Potassium channels are tetrameric membrane-spanning proteins that provide a selective pore for the conduction of K(+) across the cell membranes. One of the main physiological functions of potassium channels is efficient and very selective transport of K(+) ions through the membrane to the cell. Classical views of ion selectivity are summarized within a historical perspective, and contrasted with the molecular dynamics (MD) simulations free energy perturbation (FEP) performed on the basis of the crystallographic structure of the KcsA phospholipid membrane. The results show that the KcsA channel does not select for K(+) ions by providing a binding site of an appropriate (fixed) cavity size. Rather, selectivity for K(+) arises directly from the intrinsic local physical properties of the ligands coordinating the cation in the binding site, and is a robust feature of a pore symmetrically lined by backbone carbonyl groups. Further analysis reveals that it is the interplay between the attractive ion-ligand (favoring smaller cation) and repulsive ligand-ligand interactions (favoring larger cations) that is the basic element governing Na(+)/K(+) selectivity in flexible protein binding sites. Because the number and the type of ligands coordinating an ion directly modulate such local interactions, this provides a potent molecular mechanism to achieve and maintain a high selectivity in protein binding sites despite a significant conformational flexibility.


Determination of ²³⁷Np in environmental and nuclear samples: a review of the analytical method.
A number of analytical methods has been developed and used for the determination of neptunium in environmental and nuclear fuel samples using alpha, ICP-MS spectrometry, and other analytical techniques. This review summarizes and discusses development of the radiochemical procedures for separation of neptunium (Np), since the beginning of the nuclear industry, followed by a more detailed discussion on recent trends in the separation of neptunium. This article also highlights the progress in analytical methods and issues associated with the determination of neptunium in environmental samples.


Topographical display and interpretation of event-related desynchronization during a visual-verbal task.
Multichannel EEG recordings were performed during a visual-verbal judgement task with a verbal response using an event-related paradigm. EEG trials of 7 sec were first digitally band pass filtered (10-12 Hz, 10-11 Hz, 11-12 Hz), and the amplitudes were then squared and averaged over all trials. This processing method results in a time course of alpha power and allows us to quantify the event-related desynchronization (ERD) in each EEG channel and to compute series of ERD maps in intervals of 125 msec. Analyzing the 10-11 Hz band, it was possible to study the time course and topographical pattern of cortical activation during visual encoding and cognition. Analysis of the 11-12 Hz band has enabled us, for the first time, to study the time course, spatial localization and extent of activation of speech, premotor and motor areas. We found that the speech centers were activated maximally 250 to 375 msec before speech onset and the SMA about 250 msec before speech onset. The results are preliminary, but demonstrate how much information can be extracted from the scalp EEG.


An update on nonsteroidal anti-inflammatory drugs (NSAIDs) in small animals.
There are several choices of nonsteroidal anti-inflammatory drugs (NSAIDs) for treating dogs that have osteoarthritis. However, fewer drugs are available for cats. Like people, there may be greater differences among individuals in their response than there are differences among the drugs. In past practice, veterinarians often selected aspirin or phenylbutazone as an initial drug, and then progressed to off-label human drugs or other agents as an alternative. Now we have the advantage of several approved NSAIDs for which there are excellent published studies and US Food and Drug Administration or foreign approval to guide clinical use and safe dosages.


Decadal forest dynamics in logged and unlogged sites at Uppangala, Western Ghats, India.
Selective logging disrupts forests, changing their structure and species composition. Long-term monitoring helps in identifying the factors influencing it and aids in designing management plans. We conducted a quantitative re-assessment of trees ≥ 30 cm girth at breast height in four 1 ha plots in logged and two 1 ha plots in adjacent unlogged compartments of Uppangala forest continuum in the Western Ghats, India to compare the structural and compositional changes after a decade (2010-2021). Altogether, four species disappeared and three species were newly recruited. Mean species richness and stem density of both the forest sites decreased. Logged plots showed a slight increase in basal area (2.5%) and biomass (5.1%), whereas unlogged plots showed a decline in basal area (3.92%) and biomass (2.9%). As compared to unlogged plots, all the demographic rates were higher for logged forest sites. Across the six individual plots, the growth rates varied significantly owing to wood density and forest strata categories. Non-metric multidimensional scaling (NMDS) identified three groups with significant difference in species composition, where logged and unlogged plots have a distinct composition except for one plot. Although species richness and stem diversity remained stable, the species composition is different 37 years after logging, and the impacts of logging are still evident in the forest.


Occurrence of oligosaccharides in feces of breast-fed babies in their first six months of life and the corresponding breast milk.
The characterization of oligosaccharides in the feces of breast-fed babies is a valuable tool for monitoring the gastrointestinal fate of human milk oligosaccharides (HMOs). In the present study we monitored fecal oligosaccharide profiles together with the HMO-profiles of the respective breast milks up to six months postpartum, by means of capillary electrophoresis-laser induced fluorescence detection and mass spectrometry. Eleven mother/child pairs were included. Mother's secretor- and Lewis-type included all combinations [Le(a-b+), Le(a+b-), Le(a-b-)]. The fecal HMO-profiles in the first few months of life are either predominantly composed of neutral or acidic HMOs and are possibly effected by the HMO-fingerprint in the respective breast milk. Independent of the initial presence of acidic or neutral fecal HMOs, a gradual change to blood-group specific oligosaccharides was observed. Their presence pointed to a gastrointestinal degradation of the feeding-related HMOs, followed by conjugation with blood group specific antigenic determinants present in the gastrointestinal mucus layer. Eleven of these 'hybrid'-oligosaccharides were annotated in this study. When solid food was introduced, no HMOs and their degradation- and metabolization products were recovered in the fecal samples.


Ureaplasma urealyticum and chronic lung disease.
Neonatal lower respiratory tract colonisation with mycoplasma organisms was examined for an association with chronic lung disease. Ureaplasma urealyticum colonised 9/70 (13%) infants less than 1500 g. Seven (78%) colonised and 33 (54%) non-colonised infants developed chronic lung disease. Logistic regression analyses revealed early gestation, but not mycoplasma colonisation, was independently associated with chronic lung disease.


Host-vs.-altered-host eruptions in patients on liposomal doxorubicin.
Anthracycline antitumor antibiotics are the most commonly used chemotherapeutic agents. One of these is doxorubicin. Liposomal doxorubicin was developed as a drug delivery system in order to deliver the active drug intracellularly while decreasing the systemic toxicity, particularly hematological and cardiac toxicity. The clinical and histologic findings of the cutaneous eruptions of associated with liposomal doxorubicin are reviewed. The eruptions occurred in three women with metastatic ovarian carcinoma who were treated with liposomal doxorubicin. These three patients developed erythematous macular/papular to plaque cutaneous lesions, and in one patient a vesicular component. The eruptions involved the trunk and extremities approximately 3-4 weeks after completions of therapy. None of the patients had any documented infections, and none of the patients had symptoms other than pruritus. The eruptions cleared over a period of weeks to months. Histologic features included an interface dermatitis with numerous apoptotic/dyskeratotic cells within the epidermis, with involvement the intra-epidermal sweat ducts and the infundibulum of hair follicles. We believe that these eruptions represented a chemotherapy induced host-vs.-altered host reaction.


Assailant and victim behaviors immediately preceding inpatient assault.
The goals of this study were to detect and define victim and assailant behaviors immediately before inpatient assault and to determine their specificity to assault, to assailants, and to more serious assaults. Assault was defined as physical contact such as hitting and slapping and detected using video-cameras installed on a specialized ward for violent patients. Antecedent behaviors, or cues, were identified in victim and assailant in the five minutes preceding assault and in control periods, five minute segments in which both patients were present but no assault occurred. Fifty-five assaults between 59 patients were detected. Threatening and intrusive behaviors in assailant and victim preceded 60% of assaults and 10% of control periods. They were more than ten times more numerous before assaults. Assault was often the end of a set of overt, recognizable interactions between patients. In a chronically ill inpatient population these behaviors can be useful predictors of assault.


Radiotherapy-Induced Neurocognitive Dysfunction in Brain Tumor Survivors: Burden and Rehabilitation.
Radiotherapy-induced neurocognitive dysfunction after cranial irradiation has an incidence of 40-100%. It may affect both children and adults, and represents a significant burden not only on ill individuals and their caregivers but also on the health care system and society in general. Multiple patient-, tumor-, and treatment-related factors may contribute to development of this complication, but its pathophysiological mechanisms are still not understood clearly. It is hoped that introduction of more advanced techniques for conformal irradiation, optimized dosimetry, and specific prophylactic measures will decrease the risk of neurocognitive decline in brain tumor survivors in the future.


Transgene-free direct conversion of murine fibroblasts into functional muscle stem cells.
Transcription factor-based cellular reprogramming provides an attractive approach to produce desired cell types for regenerative medicine purposes. Such cellular conversions are widely dependent on viral vectors to efficiently deliver and express defined factors in target cells. However, use of viral vectors is associated with unfavorable genomic integrations that can trigger deleterious molecular consequences, rendering this method a potential impediment to clinical applications. Here, we report on a highly efficient transgene-free approach to directly convert mouse fibroblasts into induced myogenic progenitor cells (iMPCs) by overexpression of synthetic MyoD-mRNA in concert with an enhanced small molecule cocktail. First, we performed a candidate compound screen and identified two molecules that enhance fibroblast reprogramming into iMPCs by suppression of the JNK and JAK/STAT pathways. Simultaneously, we developed an optimal transfection protocol to transiently overexpress synthetic MyoD-mRNA in fibroblasts. Combining these two techniques enabled robust and rapid reprogramming of fibroblasts into Pax7 positive iMPCs in as little as 10 days. Nascent transgene-free iMPCs proliferated extensively in vitro, expressed a suite of myogenic stem cell markers, and could differentiate into highly multinucleated and contractile myotubes. Furthermore, using global and single-cell transcriptome assays, we delineated gene expression changes associated with JNK and JAK/STAT pathway inhibition during reprogramming, and identified in iMPCs a Pax7+ stem cell subpopulation resembling satellite cells. Last, transgene-free iMPCs robustly engrafted skeletal muscles of a Duchenne muscular dystrophy mouse model, restoring dystrophin expression in hundreds of myofibers. In summary, this study reports on an improved and clinically safer approach to convert fibroblasts into myogenic stem cells that can efficiently contribute to muscle regeneration in vivo.


The 70 kD component of the heart sarcolemmal Na+/Ca(2+)-exchanger preparation is the C-terminal portion of the protein.
The cardiac sarcolemmal Na+/Ca(2+)-exchanger was expressed in COS-7 cells by the vaccinia virus system as a fusion protein with a poly-His tag at its C-terminus. Extracts of cells expressing the exchanger construct without the His-tag reacted with an antiserum against the C-terminal portion of the main intracellular loop of the exchanger: in agreement with the finding routinely made on heart sarcolemma and on plasma membrane of cells expressing the cardiac exchanger gene, three bands of about 160, 120, and 70 kD were detected in Western blots. All three bands shifted to higher molecular masses when the construct with the His-tag was expressed, indicating that the three proteins had the same C-terminus. Thus, the 70 kD protein, whose nature has always been obscure, appears to be a degradation product of one of the two larger proteins. N-terminal sequencing of partially purified exchanger preparations has identified the cleavage site(s) producing the 70 kD protein in the 257-269 residue region of the exchanger molecule.


Electronic nicotine delivery systems (ENDS): A convenient means of smoking?
Electronic nicotine delivery systems (ENDS), which are becoming increasingly popular in many parts of the world, have recently become more sophisticated in terms of their more active content and better controlled vaporisation. This review begins by describing how cigarette smoking led to the development of ENDS as a means of combatting nicotine addiction. ENDS are usually categorised as belonging to one of only three main generations, but a fourth has been added in order to differentiate the latest, most powerful, most advanced and innovative that have improved heating efficiency. Descriptions of the principal substances contained in ENDS are followed by considerations concerning the risk of toxicity due to the presence of albeit low concentrations of such a variety of compounds inhaled over a long time, and the increasingly widespread use of ENDS as a means of smoking illicit drugs. We also review the most widely used pharmacotherapeutic approaches to smoking cessation, and recent epidemiological data showing that ENDS can help some people to stop smoking. However, in order to ensure their appropriate regulation, there is a need for higher-quality evidence concerning the health effects and safety of ENDS, and their effectiveness in discouraging tobacco smoking.


Isolation of two novel candidate hormones using a chemical method for finding naturally occurring polypeptides.
Naturally occurring peptides with biological actions have in most cases been detected by observing their biological activities in crude extracts and their isolation has been followed using bioassays. As a complement to the classical biological detection systems, we have proposed a chemical detection system based on fragmentation of peptides in tissue extracts followed by identification of certain of these peptide fragments having distinct chemical features. One such chemical feature is the C-terminal amide structure which is characteristic of many biologically active peptides. We have devised a chemical assay method for peptides having such a structure and have found several previously unknown peptide amides in procine upper small intestinal tissues. We report here the isolation and characterization of two of them, designated PHI and PYY. PHI is related to secretin, vasoactive intestinal polypeptide (VIP, glucagon and gastric inhibitory polypeptide (GIP); PYY is related to the pancreatic polypeptide and to neurotensin. Both peptides exhibit biological activities and appear to be present not only in the intestine but also in brain.


Promoting optic nerve regeneration in adult mice with pharmaceutical approach.
Our previous research has suggested that lack of Bcl-2-supported axonal growth mechanisms and the presence of glial scarring following injury are major impediments of optic nerve regeneration in postnatal mice. Mice overexpressing Bcl-2 and simultaneously carrying impairment in glial scar formation supported robust optic nerve regeneration in the postnatal stage. To develop a therapeutic strategy for optic nerve damage, the combined effects of chemicals that induce Bcl-2 expression and selectively eliminate mature astrocytes--scar forming cells--were examined in mice. Mood-stabilizer, lithium, has been shown to induce Bcl-2 expression and stimulate axonal outgrowth in retinal ganglion cells in culture and in vivo. Moreover, astrotoxin (alpha-aminoadipate), a glutamate analogue, selectively kills astrocytes while has minimal effects on surrounding neurons. In the present study, we sought to determine whether concurrent applications of lithium and astrotoxin were sufficient to induce optic nerve regeneration in mice. Induction of Bcl-2 expression was detected in the ganglion cell layer (GCL) of mice that received a lithium diet in compared with control-treated group. Moreover, efficient elimination of astrocytes and glial scarring was observed in the optic nerve of mice treated with astrotoxin. Simultaneous application of lithium and astrotoxin, but not any of the drugs alone, induced robust optic nerve regeneration in adult mice. These findings further support that a combinatorial approach of concurrent activation of Bcl-2-supported growth mechanism and suppression of glial scarring is required for successful regeneration of the severed optic nerve in adult mice. They suggest a potential therapeutic strategy for treating optic nerve and CNS damage.


Mutated tyrosine kinases as therapeutic targets in myeloid leukemias.
Tyrosine kinases are commonly mutated and activated in both acute and chronic myeloid leukemias. Here, we review the functions, signaling activities, mechanism of transformation, and therapeutic targeting of two prototypic tyrosine kinase oncogenes, BCR-ABL and FLT3, associated with chronic myeloid leukemia (CML) and acute myeloid leukemia (AML), respectively. BCR-ABL is generated by the Philadelphia chromosome translocation between chromosomes 9 and 22, creating a chimeric oncogene in which the BCR and c-ABL genes are fused. The product of this oncogene, BCR-ABL, has elevated ABL tyrosine kinase activity and transforms hematopoietic cells by exerting a wide variety of biological effects, including reduction in growth factor dependence, enhanced viability, and altered adhesion of chronic myelocytic leukemia (CML) cells. Elevated tyrosine kinase activity of BCR-ABL is critical for activating downstream signalling cascades and for all aspects of transformation, explaining the remarkable clinical efficacy of the tyrosine kinase inhibitor, imatinib mesylate (STI571). By comparison, FLT3 is mutated in about one third of all cases of AML, most often through a mechanism that involves an internal tandem duplication (ITD) of a small number of amino acid residues in the juxtamembrane domain of the receptor. As is the case for BCR-ABL, these mutations activate the kinase activity constitutively, activate multiple signaling pathways, and result in an augmentation of proliferation and viability. Transformation by FLT3-ITD can readily be observed in murine models, and FLT3 cooperates with other types of oncogenes to create a fully transformed acute leukemia. FLT3 tyrosine kinase inhibitors are currently being evaluated in clinical trials and may be very useful therapeutic agents in AML.


Microbiological analysis of patients with first and recurrent episodes of acute cholangitis in a middle-sized hospital: A single-center retrospective study in rural North Kyoto, Japan.
Knowledge of the bacterial spectrum involved in acute cholangitis is essential for adequate empiric antibiotic treatment. There is a lack of published data comparative data between patients with first and recurrent episodes of acute cholangitis. This study aimed to analyze the microbial spectrum in patients with first and second episodes of acute cholangitis. We retrospectively assessed 251 patients with first episodes of acute cholangitis between January 2014 to September 2020. At the first episode of acute cholangitis, the predominant strains belonged to Escherichia coli (17.9%), followed by Klebsiella spp. (15.5%), Enterobacter spp. (6.4%), and Enterococcus spp. (5.6%). During follow-up, acute cholangitis recurred in 109 patients; at the second episode, the predominant strains belonged to Enterococcus spp. (35.8%), followed by Klebsiella spp. (27.5%), Enterobacter spp. (22.9%), and Escherichia coli (15.6%). Enterococcus spp. were the most common pathogen in patients with second episode of acute cholangitis, regardless of whether the cholangitis was caused by a malignant tumor or a benign disease. Unlike in patients with a first episode of acute cholangitis, clinicians should consider empirical treatment with anti-enterococcal antibiotics in patients with recurrent episodes of acute cholangitis.


Continuous versus routine EEG in patients after cardiac arrest: Analysis of a randomized controlled trial (CERTA).
Electroencephalography (EEG) is essential to assess prognosis in patients after cardiac arrest (CA). Use of continuous EEG (cEEG) is increasing in critically-ill patients, but it is more resource-consuming than routine EEG (rEEG). Observational studies did not show a major impact of cEEG versus rEEG on outcome, but randomized studies are lacking. We analyzed data of the CERTA trial (NCT03129438), including comatose adults after CA undergoing cEEG (30-48 hours) or two rEEG (20-30 minutes each). We explored correlations between recording EEG type and mortality (primary outcome), or Cerebral Performance Categories (CPC, secondary outcome), assessed blindly at 6 months, using uni- and multivariable analyses (adjusting for other prognostic variables showing some imbalance across groups). We analyzed 112 adults (52 underwent rEEG, 60 cEEG,); 31 (27.7%) were women; 68 (60.7%) patients died. In univariate analysis, mortality (rEEG 59%, cEEG 65%, p = 0.318) and good outcome (CPC 1-2; rEEG 33%, cEEG 27%, p = 0.247) were comparable across EEG groups. This did not change after multiple logistic regressions, adjusting for shockable rhythm, time to return of spontaneous circulation, serum neuron-specific enolase, EEG background reactivity, regarding mortality (cEEG vs rEEG: OR 1.60, 95% CI 0.43-5.83, p = 0.477), and good outcome (OR 0.51, 95% CI 0.14-1.90, p = 0.318). This analysis suggests that cEEG or repeated rEEG are related to comparable outcomes of comatose patients after CA. Pending a prospective, large randomized trial, this finding does not support the routine use of cEEG for prognostication in this setting. Continuous EEG Randomized Trial in Adults (CERTA); NCT03129438; July 25, 2019.


Tubb3 regulation by the Erk and Akt signaling pathways: a mechanism involved in the effect of arginine ADP-ribosyltransferase 1 (Art1) on apoptosis of colon carcinoma CT26 cells.
The influence of the most important classical mono-ADP-ribosyltransferase, arginine ADP-ribosyltransferase 1 (Art1), on survival and apoptosis of colon carcinoma cells and the potential mechanisms have been partly discussed in our previous study but still need to be further studied. In this present study, Art1 of colon carcinoma CT26 cells was silenced with lentiviral vector-mediated short hairpin RNA (shRNA) or overexpressed with lentiviral vector-mediated complementary DNA (cDNA) and allograft transplant tumors are established in Balb/c mice. We verified Art1 knockdown increases apoptosis of CT26 cells transplant tumor; Art1 overexpression acts oppositely. Accordingly, growth of transplant tumors is inhibited in Art1 knockdown transplant tumors and increases in Art1 overexpression transplant tumors. Furthermore, activity of Akt and Erk cell signal pathways and expression of an apoptosis biomarker, βIII-tubulin (Tubb3), decrease when Art1 was silenced and increase when Art1 was overexpressed. Inhibiting Akt pathway or Erk pathway both downregulates expression of Tubb3 on protein and messenger RNA (mRNA) level, indicating that Tubb3 could be regulated by both Akt and Erk pathways, and plays a role in the influence of Art1 on apoptosis of Balb/c mice allograft transplant tumor. We also demonstrated that Bcl-2 family is not the responsible downstream factor of the Erk pathway in colon carcinoma cells which is undergoing apoptosis. These findings enrich the molecular mechanism for the function of Art1 in colon carcinoma and provide a complementary support for Art1 to be a potential therapeutic target of the treatment of this kind of malignant tumor.


Inefficient licking during forced spout alternation in rats: violation of the law of effect?
The efficiency of the neural control of licking was assessed in rats (n = 6) allowed 30-min daily access to water in a box equipped with two retractable drinking spouts. After completion of a photoelectrically monitored lick, the contacted spout was withdrawn by an electromechanical device which simultaneously made the other spout available at an adjacent wall opening. Although economy of behavior required the animals to lick at each spout only once, computer monitored licking showed that the rats emitted 2.9 +/- 0.2 and 2.5 +/- 0.1 licks per spout presentation (LPSP) on the first and fifth days of training, respectively. The transition time between spouts and the interlick interval were not significantly changed. Separation of the spouts by a vertical partition extending 24 mm into the box prolonged the transition time and increased LPSP to 3.9 and 3.3 on Days 1 and 4 of training, respectively. These values were not changed when the length of the partition was increased to 40 mm. The failure of rats to maximize reward by emitting single licks probably reflects limited control of the generator of licking and/or the tendency to avoid its frequent switching on and off.


An integrated multifunctional hybrid cement (pRMGIC) for dental applications.
Glass-ionomer and resin-modified glass-ionomer cements are versatile materials with the ability to form a direct bond with tooth tissues. The aim of this study was to formulate a novel class of dental bio-interactive restorative material (pRMGIC) based on resin-modified glass-ionomer cements via the inclusion of an organophosphorus monomer, ethylene glycol methacrylate phosphate, with a potential to improve the mechanical properties and also function as a reparative restorative material. pRMGIC was formulated with modification of the resin phase by forming mixes of ethylene glycol methacrylate phosphate (EGMP; 0-40%wt) and 2-hydroxyethyl methacrylate monomer into the liquid phase of a RMGIC (Fuji II LC, GC Corp.). The physical properties of the cements were determined including setting characteristics, compressive strength and modulus (CS &CM), microhardness (MH) and biaxial flexural strength (BFS). Fluid uptake and fluoride release were assessed up to 60 days storage. Adhesion to sound dentine was measured using micro-tensile bond strength and surface integrity was analysed using SEM coupled with EDX. Statistical analysis was performed using ANOVA and Bonferroni post-hoc tests. The pRMGIC cements exhibited an increase in working time with increasing EGMP concentration however were within the limits of standard clinical requirements. Although the compressive strength of pRMGIC cements were comparable to control cements in the early stages of maturation, the higher EGMP-containing cements (EGMP30 and 40) exhibited significantly greater values (p < 0.05) after 4 weeks storage (141.0 ± 9 and 140.4 ± 8 MPa, respectively), in comparison to EGMP0 (128.8 ± 7 MPa). A dramatic two fold increase in biaxial flexural strength (p < 0.001) was observed for the pRMGIC's. Furthermore, the ability to decalcify tooth apatite resulted in enhanced interfacial adhesion due to chelation with calcium ions of tooth apatite. The inclusion of EGMP encouraged formation of reinforcing complexes within the RMGIC, thus improving physical properties, decreasing solubility and lower fluoride release. A dense microstructure was observed with increasing EGMP content. A novel universal bio-interactive adhesive repair material will enable clinicians to offer more effective repair of the tooth-restoration complex, thus future treatments will benefit both patient and a severely constrained healthcare budget.


[The effect of etimizol on the functional status of the blood coagulation systems and fibrinolysis in workers on expedition-duty watches working under the climatic conditions of northern Tyumen Province].
The study was undertaken to reveal the effects of ethymisole on the functional status of blood coagulation and fibrinolytic systems, the body's adaptative reserves of workers from expeditionary-duty teams. Ethymisole was found to normalize the functional status of blood coagulation and fibrinolytic systems, to restore conjunction of their functioning. The use of the drug was ascertained to normalize the activity of lipid peroxidation products, to enhance the adaptative reserves of expeditionary-duty workers by prolonging the resistance phase. It is concluded that ethymisole may be used as an agent that enhances the body's adaptative reserves.


Vocalizations associated with pectoral fin contact in bottlenose dolphins (Tursiops truncatus).
Pectoral fin contact in bottlenose dolphins represents one form of tactile communication. Acoustic communication associated with pectoral fin contact is an additional level of communication that may change or enhance the tactile message between two individuals. In this study, we examine vocalization types associated with pectoral fin contact in a group of captive bottlenose dolphins (Tursiops truncatus). From 2006 to 2009, vocalizations potentially associated with 748 pectoral fin contacts were examined: whistles, click trains and overlap of whistles and click trains were documented when associated with fin contact. Dolphins were also documented not vocalizing when exchanging pectoral fin contacts. Call type associated with pectoral fin contact was compared for the proportion of the type of pectoral fin contact, vocalizer sex, initiator and receiver roles, and gender pair. Overall, vocalizations differed significantly by vocalizer role as rubber or rubbee, initiator, and sex. Receivers and rubbees clicked and used overlap vocalizations more frequently, and males produced overlap vocalizations more frequently. These results suggest that whistles may be used to initiate pectoral fin contact or show preference for a particular partner, while click trains may be used to show disinterest in pectoral fin contact or to signal the end of a contact. Examining vocalizations produced in conjunction with tactile contact is a relatively new approach in the study of individual dolphin behavior and may be useful for understanding dolphin social alliances and social preferences for various individuals within a population.


Use and misuse of random forest variable importance metrics in medicine: demonstrations through incident stroke prediction.
Machine learning tools such as random forests provide important opportunities for modeling large, complex modern data generated in medicine. Unfortunately, when it comes to understanding why machine learning models are predictive, applied research continues to rely on 'out of bag' (OOB) variable importance metrics (VIMPs) that are known to have considerable shortcomings within the statistics community. After explaining the limitations of OOB VIMPs - including bias towards correlated features and limited interpretability - we describe a modern approach called 'knockoff VIMPs' and explain its advantages. We first evaluate current VIMP practices through an in-depth literature review of 50 recent random forest manuscripts. Next, we recommend organized and interpretable strategies for analysis with knockoff VIMPs, including computing them for groups of features and considering multiple model performance metrics. To demonstrate methods, we develop a random forest to predict 5-year incident stroke in the Sleep Heart Health Study and compare results based on OOB and knockoff VIMPs. Nearly all papers in the literature review contained substantial limitations in their use of VIMPs. In our demonstration, using OOB VIMPs for individual variables suggested two highly correlated lung function variables (forced expiratory volume, forced vital capacity) as the best predictors of incident stroke, followed by age and height. Using an organized analytic approach that considered knockoff VIMPs of both groups of features and individual features, the largest contributions to model sensitivity were medications (especially cardiovascular) and measured medical risk factors, while the largest contributions to model specificity were age, diastolic blood pressure, self-reported medical risk factors, polysomnography features, and pack-years of smoking. Thus, we reach very different conclusions about stroke risk factors using OOB VIMPs versus knockoff VIMPs. The near-ubiquitous reliance on OOB VIMPs may provide misleading results for researchers who use such methods to guide their research. Given the rapid pace of scientific inquiry using machine learning, it is essential to bring modern knockoff VIMPs that are interpretable and unbiased into widespread applied practice to steer researchers using random forest machine learning toward more meaningful results.


DNA sequesters endogenous mRNA during preparation of crude Escherichia coli extracts for protein synthesis; use of an S60 reduces the sequestered mRNA.
High backgrounds of endogenous incorporation of amino acids into protein in crude extracts of Escherichia coli are a consequence of endogenous messenger RNA. This RNA survives the preparation by virtue of protection or sequestering by endogenous DNA. Thus, DNase treatment in the crude extract leads to the elimination of this mRNA, while DNase treatment has no effect on the purified RNA. The endogenous mRNA also appears to be physically associated with DNA on CsCl gradients, and can be largely removed along with DNA by centrifugation of extracts at 60,000g. The top layer above a 60,000g centrifugation (S60) appears to be suitable for protein synthesis, and provides for lower background levels of endogenous messenger RNA.


Molecular cloning of a cDNA sequence for rat malic enzyme. Direct evidence for induction in vivo of rat liver malic enzyme mRNA by thyroid hormone.
Malic enzyme (EC 126.96.36.199) mRNA was partially purified to 12% of total mRNA activity (greater than 150-fold enrichment) from 3-5-3'-triiodo-L-thyronine-carbohydrate-stimulated rat liver by polysome immunopurification followed by oligo(dT)-cellulose chromatography. This preparation was used as the template for synthesis of cDNA on a pBR322-SV40 hybrid plasmid DNA primer which was then circularized with linker DNA and used to transform Escherichia coli. The resulting transformants were screened by in situ differential hybridization using 32P-labeled poly(A+)RNA prepared from uninduced and 3-5-3'-triiodo-L-thyronine-carbohydrate-induced rat livers. Of 750 transformants screened, 6 were found to hybridize differentially; 1 of these, prME, contained an insert of about 1250 base pairs and hybrid-selected an mRNA which directed synthesis of malic enzyme in a cell-free translation system. Using this cDNA as a probe, we demonstrated that the level of malic enzyme mRNA after thyroid hormone treatment was markedly increased and the size of the major malic enzyme mRNA was shown by Northern analysis to be about 2700 nucleotides in length.


The Impact of Chronic Pelvic Ischemia on LUTS and Urinary Levels of Neuroinflammatory, Inflammatory, and Oxidative Stress Markers in Elderly Men: A Case-control Study.
To investigate lower urinary tract symptoms (LUTS) and urinary levels of neuroinflammatory, inflammatory, and oxidative stress markers in elderly men with chronic pelvic ischemia (CPI) caused by significant aortoiliac disease. Thirteen men aged over 60 years, with aorta, unilateral or bilateral common/internal iliac artery occlusion documented by computed tomography angiography or angiography, were enrolled from the vascular surgery department. Twelve sex- and age-matched controls without significant aortoiliac disease were used for comparison. Exclusion criteria included neurogenic bladder dysfunction, bladder or prostate cancer, prostatic surgery, pelvic radiotherapy, or chronic treatment for LUTS. Participants underwent urological examination, including assessment of International Prostate Symptom Score (IPSS), uroflowmetry, postvoid residual (PVR), and prostate volume. Urine samples were collected, and levels of neuroinflammatory (nerve growth factor, NGF), inflammatory (cytokines), and oxidative stress markers (8-hydroxy-2'-deoxyguanosine) were determined by enzyme-linked immunosorbent assay. Groups were similar for age, PVR, prostate volume, and most cardiovascular risk factors. IPSS was higher in patients with CPI (11 ± 3 vs 8 ± 2, P = .02), with a significant mean difference between groups of three points. Urinary NGF was significantly higher in men with CPI (3.7 ± 0.8 vs 2.9 ± 0.7, P = .02), but no differences were found in inflammatory and oxidative biomarkers among groups. Severe CPI in elderly men is associated with a significant increase in LUTS and bladder neurogenic inflammation, as suggested by the increase of NGF release in urine, sensitizing bladder afferents. These findings confirm the relevance of ischemia in bladder function and appear to validate animal models of bilateral iliac artery occlusion.


Sentinels of chromatin: chromodomain helicase DNA-binding proteins in development and disease.
Chromatin is highly dynamic, undergoing continuous global changes in its structure and type of histone and DNA modifications governed by processes such as transcription, repair, replication, and recombination. Members of the chromodomain helicase DNA-binding (CHD) family of enzymes are ATP-dependent chromatin remodelers that are intimately involved in the regulation of chromatin dynamics, altering nucleosomal structure and DNA accessibility. Genetic studies in yeast, fruit flies, zebrafish, and mice underscore essential roles of CHD enzymes in regulating cellular fate and identity, as well as proper embryonic development. With the advent of next-generation sequencing, evidence is emerging that these enzymes are subjected to frequent DNA copy number alterations or mutations and show aberrant expression in malignancies and other human diseases. As such, they might prove to be valuable biomarkers or targets for therapeutic intervention.


Lamotrigine reduces spontaneous and evoked GABAA receptor-mediated synaptic transmission in the basolateral amygdala: implications for its effects in seizure and affective disorders.
Lamotrigine (LTG) is an antiepileptic drug that is also effective in the treatment of certain psychiatric disorders. Its anticonvulsant action has been attributed to its ability to block voltage-gated Na(+) channels and reduce glutamate release. LTG also affects GABA-mediated synaptic transmission, but there are conflicting reports as to whether inhibitory transmission is enhanced or suppressed by LTG. We examined the effects of LTG on GABA(A) receptor-mediated synaptic transmission in slices from rat amygdala, a brain area that is particularly important in epileptogenesis and affective disorders. In intracellular recordings, LTG (100 microM) reduced GABA(A) receptor-mediated IPSPs evoked by electrical stimulation in neurons of the basolateral nucleus. In whole-cell recordings, LTG (10, 50 and 100 microM) decreased the frequency and amplitude of spontaneous IPSCs, as well as the amplitude of evoked IPSCs, but had no effect on the kinetics of these currents. LTG also had no effects on the frequency, amplitude or kinetics of miniature IPSCs recorded in the presence of TTX. These results suggest that in the basolateral amygdala, LTG suppresses GABA(A) receptor-mediated synaptic transmission by a direct and/or indirect effect on presynaptic Ca(++) influx. The modulation of inhibitory synaptic transmission may be an important mechanism underlying the psychotropic effects of LTG.


Disappearance of dead plant material in a mixed grass prairie.
Disappearance of dead plant material was studied in ungrazed mixed grass prairie in Western North Dakota. Disappearance rates varied both within and between years as well as with method used. The highest rates (2.07 to 3.15 g/m2/day) were recorded early in the season and disappearance continued during the winter but at a slower rate (0.99 g/m2/day). Some differences were found in rates of disappearance of dead material of different species during the first two months. Annual decomposition rates ranged from 400 g/m2 to 526 g/m2. The ash content of dead material showed generally increasing values with advancing decomposition. The turnover time of above-ground biomass appeared to be approximately three years.


Post infectious CNS disorders: towards a unified approach.
The spectrum of post infectious (PI) central nervous system (CNS) conditions includes a range of grey and white-matter disorders which can occur after viral or bacterial infections or in response to vaccinations. The clinical, radiological and immunological phenomenology raises a number of issues regarding the nature of immune-mediated CNS abnormalities, their etiology, pathogenesis and therapy. Here we focus on crucial issues pertaining to pathogenesis and aim to identify where current knowledge is insufficient in order to suggest future avenues of clinical and experimental research that may help to devise optimal therapy for these conditions.


Effects of administration of testosterone on seminal vesicle activity in the catfish Clarias batrachus during preparatory phase: a study correlating changes in testis and serum sex steroid levels.
In the catfish Clarias batrachus administration of testosterone (0.25, 0.5 and 1.0 microgram/g BW for 30 days) during the preparatory phase increased significantly gonadosomatic index (GSI), seminal vesicle-somatic index (SV-SI) and concentrations of proteins, fructose, hexosamines, and sialic acid in the SV and testis in a dose-dependent manner. These stimulatory effects could be correlated positively with the increase of serum testosterone level. Serum estradiol-17 beta was also significantly high in the testosterone groups.


Caesarean section combined with temporary aortic balloon occlusion followed by uterine artery embolisation for the management of placenta accreta.
To determine the efficacy and safety of caesarean section combined with temporary aortic balloon occlusion followed by uterine artery embolisation (UAE) for the treatment of patients with placenta accreta. This retrospective study involved 42 patients with placenta accreta. All patients underwent caesarean section combined with temporary aortic balloon occlusion followed by UAE through the right femoral approach. All patients were confirmed to have placenta praevia and accreta, including five patients with placenta percreta, at the time of delivery. The technical success rate of the combined treatment was 97.6% (41/42). Forty-one patients underwent successful caesarean section with conservation of the uterus. Hysterectomy was required in one (3.1%) patient. The amount of blood loss and blood transfusion, and the operation time were was 586 ± 355 ml, 422 ± 83 ml and 65.5 ± 10.6 minutes, respectively. The mean postoperative hospital stay, occlusion time and fetal radiation dose were 5.5 ± 2.6 days, 22.4 ± 7.2 minutes and 4.2 ± 2.9 mGy, respectively. There were no significant changes before and 7 days after the endovascular procedure in creatinine levels (56.8 ± 13.8 μmol/l versus 63.4 ± 16.7 μmol/l, p = 0.09) or urea nitrogen (6.3 ± 2.5 μmol/l versus 7.4 ± 3.8 μmol/l, p = 0.17). There were no access-site complications after the endovascular procedure and no complications related to the intervention during follow-up. Temporary aortic balloon occlusion followed by UAE can effectively control postpartum haemorrhage during placental dissection, and reduce transfusion requirements, hysterectomy rate, and operation time in patients with placenta accreta.


Evaluating whether direct-to-consumer marketing can increase demand for evidence-based practice among parents of adolescents with substance use disorders: rationale and protocol.
Fewer than one in 10 adolescents with substance use disorders (ASUDs) will receive specialty treatment, and even fewer will receive treatment designated as evidence-based practice (EBP). Traditional efforts to increase the utilization of EBP by ASUDs typically focus on practitioners-either in substance use clinics or allied health settings. Direct-to-consumer (DTC) marketing that directly targets parents of ASUDs represents a potentially complementary paradigm that has yet to be evaluated. The current study is the first to evaluate the relevance of a well-established marketing framework (the Marketing Mix) and measurement approach (measurement of perceived service quality [PSQ]) with parents of ASUDs in need of treatment. A mixed-methods design is employed across three study phases, consistent with well-established methods used in the field of marketing science. Phase 1 consists of formative qualitative research with parents (and a supplementary sample of adolescents) in order to evaluate and potentially adapt a conceptual framework (Marketing Mix) and measure of PSQ. Phase 2 is a targeted survey of ASUD parents to elucidate their marketing preferences, using the adapted Marketing Mix framework, and to establish the psychometric properties of the PSQ measure. The survey will also gather data on parents' preferences for different targeted marketing messages. Phase 3 is a two-group randomized controlled trial comparing the effectiveness of targeted marketing messages versus standard clinical information. Key outcomes will include parents' ratings of PSQ (using the new measure), behavioral intentions to seek out information about EBP, and actual information-seeking behavior. The current study will inform the field whether a well-established marketing framework and measurement approach can be used to increase demand for EBP among parents of ASUDs. Results of this study will have the potential to immediately inform DTC marketing efforts by professional organizations, federal agencies, clinicians, and clinical researchers.


Compaction of, and drug release from, coated drug pellets mixed with other pellets.
Tablets have been prepared at known compaction force from three types of pellets in different proportions: (1) pellets containing 80% theophylline as a model drug coated with different thicknesses of a polymer film coat, (2) pellets containing glyceryl monostearate as a deformable material, and (3) pellets containing a disintegrant. The breaking load, friability, disintegration and drug release properties were evaluated, the latter as a mean dissolution time (MDT) and its variance (VDT). The mechanism of dissolution was assessed from the value of the relative dispersion (RD) of the mean dissolution time. The possible relationship between the properties of the pellets and those of the tablets was evaluated by canonical analysis followed by multiple regression analysis. It was found that only about 51% of the tablet properties could be predicted from the properties of the pellets. The quantitative relationship between pellet properties and tablet properties was found to vary in type and level of quality. Reduction of breaking load, friability and disintegration was less predictable than that of dissolution represented as the MDT. The values of RD for the different preparations clearly identified those preparations where the film coat still retained control of the dissolution process. Such formulations contained at least 40% of soft pellets and coated pellets with at least a weight gain of 8%.


Mobile Health Applications and Medication Adherence of Patients With Hypertension: A Systematic Review and Meta-Analysis.
Current evidence has revealed the beneficial effects of mobile health applications on systolic and diastolic blood pressure. However, there is still no solid evidence of the underlying factors for these outcomes, and hypertension treatment is performed mainly by medication intake. This study aims to analyze the impacts of health applications on medication adherence of patients with hypertension and understand the underlying factors. A systematic review and meta-analysis were conducted considering controlled clinical trials published, without year filter, through July 2020. The searches were performed in the electronic databases of Scopus, MEDLINE, and BVSalud. Study characteristics were extracted for qualitative synthesis. The meta-analysis examined medication-taking behavior outcomes using the generic inverse-variance method to combine multiple variables. A total of 1,199 records were identified, of which 10 studies met the inclusion criteria for qualitative synthesis, and 9 met the criteria for meta-analysis with 1,495 participants. The analysis of mean changes revealed significant improvements in medication adherence (standardized mean difference=0.41, 95% CI=0.02, 0.79, I2=82%, p=0.04) as well as the analysis of the values measured after follow-up (standardized mean difference=0.60, 95% CI=0.30, 0.90, I2=77%, p<0.0001). Ancillary improvements were also identified, such as patients' perceived confidence, treatment self-efficacy and self-monitoring, acceptance of technology, and knowledge about the condition and how to deal with health issues. There is evidence that mobile health applications can improve medication adherence in patients with hypertension, with broad heterogeneity between studies on the topic. The use of mobile health applications conceivably leads to ancillary improvements inherent to better medication adherence.


Residual and passive force enhancement in skinned cardiac fibre bundles.
In skeletal muscle, steady-state force is consistently greater following active stretch than during a purely isometric contraction at the same length (residual force enhancement; RFE). Similarly, when deactivated, the force remains higher following active stretch than following an isometric condition (passive force enhancement; PFE). RFE and PFE have been associated with the sarcomere protein titin, but skeletal and cardiac titin have different structures, and results regarding RFE in cardiac muscle have been inconsistent and contradictory. Therefore, the purpose of this study was to determine if cardiac muscle exhibits RFE and PFE. Skinned fibre bundles (n = 10) were activated isometrically at a sarcomere length of 2.2 μm and actively stretched by 15% of their length. The resultant active and passive forces were compared to the corresponding forces obtained for purely isometric contractions at the long length. RFE was observed in all fibre bundles, averaging 5.5 ± 2.5% (ranging from 2.3 to 9.4%). PFE was observed in nine of the ten bundles, averaging 11.1 ± 6.5% (ranging from -2.1 to 18.7%). Stiffness was not different between the active isometric and the force enhanced conditions, but was higher following deactivation from the force-enhanced compared to the isometric reference state. We conclude that there is RFE and PFE in cardiac muscle. We speculate that cardiac muscle has the same RFE capability as skeletal muscle, and that the most likely mechanism for the RFE and PFE is the engagement of a passive structural element during active stretching.


Suppression of Inner Mitochondrial Membrane Peptidase 2-Like (IMMP2L) Gene Exacerbates Hypoxia-Induced Neural Death Under High Glucose Condition.
It is known that diabetes hyperglycemia enhances cerebral ischemia and reperfusion induced damage. We have previously shown that mutation of inner mitochondrial membrane peptidase 2-like (IMMP2L) increases brain damage caused by transient cerebral ischemia. In this study, we attempt to examine the impact of IMMP2L deficiency on an in vitro model that mimics the diabetic hypoxic conditions. Normal IMMP2L wild type and IMMP2L gene deleted HT22 cells were cultured. Hypoxia was induced under high glucose and acidic conditions with 4 h of oxygen deprivation. Cell viability was assessed by CCK-8 assay and cell death was determined using Annexin V/7-AAD assay. Superoxide production was measured using dihydroethidium staining and mitochondrial membrane potential was detected using JC-1 probe. Suppression of IMMP2L reduced the cell viability, increased the ROS production and decreased the mitochondrial membrane potential. In conclusion, our study demonstrated that deficiency of IMMP2L in cells, cultured under hypoxia, high glucose and acidic conditions, exacerbated neuronal death under a condition that mimics in vivo cerebral ischemia in diabetic condition.


Understanding crashes involving roadway objects with SHRP 2 naturalistic driving study data.
Crashes involving roadway objects and animals can cause severe injuries and property damages and are a major concern for the traveling public, state transportation agencies, and the automotive industry. This project involved an in-depth investigation of such crashes based on the second Strategic Highway Research Program (SHRP 2) Naturalistic Driving Study (NDS) data including detailed information and videos about 2,689 events. The research team conducted a variety of logistic regression analyses, complemented by Support Vector Machine (SVM) analyses and detailed case studies. The logistic regression results indicated that driver behavior/errors, involvement of secondary tasks, roadway characteristics, lighting condition, and pavement surface condition are among the factors that contributed significantly to the occurrence and/or increased severity outcomes of crashes involving roadway objects and animals. Among these factors, improper turning movements (odds ratio = 88), avoiding animal or other vehicle (odds ratio = 38), and reaching/moving object in vehicle (odds ratio = 29) particularly increased the odds of crash occurrence. Factors such as open country roadways, sign/signal violation, unfamiliar with roadway, fatigue/drowsiness, and speeding significantly increased the severity outcomes when such crashes occurred. The sensitivity analysis of the three SVM classifiers confirmed that driver behavior/errors, critical speed, struck object type, and reaction time were major factors affecting the occurrence and severity outcomes of events involving roadway objects and animals. Practical Applications: The study provides insights on risk factors influencing safety events involving roadway objects, including their occurrence and the severity outcomes. The findings allow researchers and traffic engineers to better understand the causes of such crashes and therefore develop more effective roadway- and vehicle- based countermeasures.


Evaluation of the role of ultrasonography in diagnosis of autoimmune thyroiditis in goitrous children.
To evaluate the role of thyroid ultrasound in children with autoimmune thyroiditis diagnosed either on cytopathology or by the presence of thyroid peroxidase antibodies. Children presenting for the first time to the Thyroid Clinic at the Institute of Nuclear Medicine and Allied Sciences for the complaint of goiter over a two year period (January 2005-December 2006) were studied. 695 school children (244 boys and 451 girls) aged 5-18 year were studied. Children were subjected to thyroid ultrasound, cytopathology, thyroid peroxidase antibody and thyroid function tests. Overall, 16% of goitrous children had hypoechogenicity on ultrasound, 15.2% had cytopathological evidence of thyroiditis, 10.6% had positive thyroid peroxidase antibodies and 25.2% had abnormal thyroid function tests. Subjects with hypoechogenicity had higher percentage of thyroiditis on cytopathology (41.4% vs. 10.3%; P<0.01), thyroid peroxidase antibody positivity (30.6% vs. 6.8%; P<0.01) and thyroid dysfunction (46.8% vs. 21.2%; P<0.01) than those with normal echogenicity. Thyroid USG has a useful, though limited, role in excluding thyroid disease in children. The sensitivity of echogenicity for the diagnosis of autoimmune thyroiditis in children is less than that reported in adults.


Molecular and cellular mechanisms of the effects of Propolis in inflammation, oxidative stress and glycemic control in chronic diseases.
Propolis is a sticky, resinous material gather from plants and is blended with wax and other constituents. It is reported to have anti-inflammatory, anti-oxidative and blood glucose-lowering properties. This review aims to summarise evidences for the cellular and molecular mechanism of Propolis in inflammation, oxidative stress, and glycemic control. Propolis stimulate the production and secretion of anti-inflammatory cytokines and to inhibit the production of inflammatory cytokines and due to its various antioxidant and poly-phenolic compounds may has a role in control and treating some of the chronic diseases. Most studies have shown that Propolis may affect metabolic factors including plasma insulin levels, and it has proposed that it could be used in the prevention and treatment of T2D Mellitus. In general, to demonstrate the definite effects of Propolis on chronic diseases, more studies are required using larger sample sizes and various doses of Propolis, using better characterized and standardized agents.


Influence of calcium on phosphorylation of a synaptosomal protein.
Synaptosomal proteins isolated from rat cerebral cortex were phosphorylated endogeneously in the presence of [gamma-32P]ATP. The phosphorylated proteins were found to be membrane bound by differential and density gradient centrifugation. In contrast to the phosphorylation of all synaptosomal proteins, phosphorylation of one protein (C), 41 000--43 000 daltons, was inhibited by Mg2+ and stimulated by Ca2+. In addition, the ionophores X537A and A23187, as well as papaverine, selectively enhanced phosphorylation of protein C without affecting phosphorylation of the outer proteins. Cyclic AMP did not influence the phosphorylation of protein C but markedly affected the phosphorylation of other synaptosomal proteins. It appears that the phosphorylation of protein C is stimulated by agents which trigger the release of neurotransmitters (Ca2+, X537A, A23187 and papaverine), and is inhibited by Mg2+, which inhibits release. It is proposed that the phosphorylation of protein C is related to membranal events underlying the release of neurotransmitters.


Intention to use maternal waiting home and its predictors among pregnant women in Ethiopia: systematic review and meta-analysis.
A high maternal death rate is a result of maternal delays in seeking emergency obstetric care, particularly in countries with limited resources like Ethiopia. Utilizing maternity waiting homes is a strategy to overcome geographical barriers and improve maternal and neonatal health outcomes. Pregnant women must intend to use this service in addition to it being available. Therefore, the goal of this study was to assess pregnant women's intentions to use maternity waiting homes and associated characteristics. PubMed, Google Scholar, Scopus, Science Direct, and online institutional repository homes were searched. Data were extracted using Microsoft Excel and analyzed using STATA statistical software (v. 14). Publication bias was checked by forest plot, Begg's, and Egger's tests. To look for heterogeneity, I2 was computed, and an overall estimated analysis was carried out. Subgroup analysis was done by study region, sample size, and publication. The pooled odds ratio for associated factors was also computed. Out of 258 articles assessed, 8 studies with 4111 study participants met the criteria and were included in this study. The pooled prevalence of intention to use maternity waiting home was 52.25% (95% CI 45.88-58.66), I2 = 93.8%). Amhara region had a higher intention to use maternal waiting for home prevalence (63.5%), per subgroup analysis. In studies with sample sizes higher than 5000, the usage of maternity waiting homes was less prevalent (45.2%). Between published research (52.9%) and unpublished studies (51.3%), there was no appreciable difference in the intention to use a maternity waiting home. Experience of maternity waiting home (AOR = 3.337; 95% CI 2.038-5.463), direct subjective norm (AOR = 2.763; 95% CI 1.395-5.471), and direct perceived behavioral control (AOR = 23.147; 95% CI 2.341-4.231). In Ethiopia, the intention to use maternity waiting was low. There was an intentional variation in to use of maternity waiting homes across regions of Ethiopia. Improving behavioral perception through intervention programs such as antenatal education should have been strengthened.


Cost-effectiveness of minimally invasive partial nephrectomy and percutaneous cryoablation for cT1a renal cell carcinoma.
There is growing evidence that partial nephrectomy (PN) and percutaneous cryoablation (PCA) yield comparable outcomes for patients with cT1a renal cell carcinoma (RCC), although the cost-effectiveness of both treatments still needs to be assessed. To perform a cost-effectiveness analysis of PN and PCA for patients with cT1a RCC. A decision analysis was created over a 5-year span from a healthcare payer's perspective computing expected costs and outcomes of PN and PCA in terms of quality-adjusted life-years (QALYs) and incremental cost-effectiveness (ICER). After each treatment, the following states were modelled using data from the recent literature: procedural complications, no evidence of disease (NED), local recurrence, metastases, and death from RCC- or non-RCC-related causes. Probabilistic and deterministic sensitivity analyses were performed. PCA and PN yielded health benefits of 3.68 QALY and 3.67 QALY. Overall expected costs were $20,491 and $26,478 for PCA and PN. On probabilistic sensitivity analysis, PCA was more cost-effective than PN in 84.78% of Monte Carlo simulations. PCA was more cost-effective until its complication risk was at least 38% higher than PN. PCA was more cost-effective than PN when (i) PCAs annual local recurrence risk was < 3.5% higher than that of PN in absolute values; (ii) PCAs annual metastatic risk was < 1.0% higher than that of PN; or (iii) PCAs annual cancer-specific mortality risk < 0.65% higher than that of PN. PCA remained cost-effective until its procedural cost is above $13,875. PCA appears to be more cost-effective than PN for the treatment of cT1a RCC, although the currently available evidence is of limited quality. PCA may be the better treatment strategy in the majority of scenarios varying procedural complications, recurrence, metastatic risk, and RCC-mortality in clinically plausible ranges. • For patients with cT1a RCCs, PCA yields a comparable health benefit at lower costs compared to PN, making PCA the dominant and therefore more cost-effective treatment strategy over PN. • PCA was more cost-effective than PN when (i) PCAs annual local recurrence risk was < 3.5% higher than PN in absolute values; (ii) PCAs annual metastatic risk was < 1.0% higher than PN; or (iii) PCAs annual cancer-specific mortality risk < 0.65% higher than PN. • PCA is more cost-effective than PN for the treatment of cT1a RCC, and it remained so in the majority of scenarios varying procedural complications, recurrence, metastatic risk, and RCC mortality.


Behavioral and electrophysiological recovery following cryogenic nerve injury.
Postthoracotomy pain can be reduced by cryoanalgesia of intercostal nerves. The technique involves focal freezing of peripheral nerves to interrupt pain pathways, producing immediate functional changes that recover as the nerves regenerate. To assess the time-course of functional changes that follow nerve injury, unilateral freeze lesions of sciatic nerve were induced in rats with a cryosurgical unit. The contralateral nerves were used as sham-operated controls. Following nerve injury, behavioral and electrophysiologic tests were repeated to 90 days. The acute effect of nerve injury was a decrease in behavioral measures of hind limb function (P less than 0.05), an increase in electrical threshold to elicit hind limb contraction (P less than 0.005), and an absence of stimulus-evoked compound action potential (P less than 0.005). Morphologic changes included substantial endoneurial edema associated with Wallerian degeneration. Remyelination occurred subsequently during the following 35 days. Although all physiologic measures returned toward normal, nerve conduction velocities were still much slower in the experimental group. In a second study, the long-term effects of cryogenic injury were compared with neurolytic injury with 10% procaine HCl, both of which produced a conduction velocity deficit that persisted at least 90 days after the initial injury. These behavioral and electrophysiologic results complement previous reports of morphologic deficits in the nerves including incomplete recovery of nerve fiber diameter and increased thickness of the perineurial sheath.


Update on gastroenteropancreatic neuroendocrine tumors.
The incidence gastroenteropancreatic neuroendocrine neoplasms (GEP-NENs) has dramatically risen over the last three decades, probably due to the increased detection of asymptomatic lesions. The diagnostic work-up for patients with suspected GEP-NENs is based on conventional imaging, endoscopy, pathology, and functional imaging, including 68Gallium-DOTATATE PET and 18F-FDG PET. The choice of the best treatment strategy should be based on the evaluation of tumor-related features and patient's characteristics. A conservative management, consisting of active surveillance or endoscopic resection, has been advocated for patients with small, incidentally discovered, nonfunctioning tumors without features of aggressiveness. On the other hand, surgery with lymphadenectomy, also with a minimally invasive approach, represents the gold standard for the curative treatment of localized disease. Moreover, surgical resection plays an important role also in the context of a multimodal treatment strategy for patients with advanced GEP-NENs. Finally, a wide range of medical therapies, comprising somatostatin analogues, peptide receptor radionuclide therapy, target therapies and several chemotherapy regimens, can be offered to patients with advanced GEP-NENs not amenable of surgical resection, according to the biological and molecular features of their disease.


Inoculation of chicks with viable non-colonizing strains of Campylobacter jejuni: evaluation of protection against a colonizing strain.
We have treated chicks with viable non-colonizing mutant strains of Campylobacter jejuni to test these as a possible vaccine. We found that intramuscular inoculation with and without adjuvant, and with or without a concomitant oral dose of non-colonizing strains, failed to provoke protective immunity.


[Statistical Mistakes Commonly Made When Writing Medical Articles].
Statistical analysis is an essential component of the medical writing process for research-related articles. Although the importance of statistical testing is emphasized, statistical mistakes continue to appear in journal articles. Major statistical mistakes can occur in any of the three different stages of medical writing, including in the design stage, analysis stage, and interpretation stage. In the design stage, mistakes occur if there is a lack of specificity regarding the research hypothesis or data collection and analysis plans. Discrepancies in the analysis stage occur if the purpose of the study and characteristics of the data are not sufficiently considered, or when an inappropriate analytic procedure is followed. After performing the analysis, the results are interpreted, and an article is written. Statistical analysis mistakes can occur if the underlying methods are incorrectly written or if the results are misinterpreted. In this paper, we describe the statistical mistakes that commonly occur in medical research-related articles and provide advice with the aim to help readers reduce, resolve, and avoid these mistakes in the future.


Fast and easy method to culture and obtain large populations of male nematodes.
Caenorhabditis elegans is a model system widely used in fundamental research. Even though, nematodes are easy to maintain in the laboratory, obtaining large populations of worms require a lot of work and is time consuming. Furthermore, because C. elegans are mainly hermaphrodite it is even more complicated to obtain large amounts of males which make high-throughput experiments using C. elegans males very challenging. In order to overcome these limitations, we developed affordable and rapid methods to: (1) grow large synchronous worm populations (2) easily obtain large amounts of males We developed a culture method on plates to grow big synchronized worm populations with the standard incubators used on all worm labs. We also established an easy filtration method allowing to obtain large male populations in an hour. After filtering, the worm population contains more than 90% of adult males and no adult hermaphrodites since all the contaminants are larva and embryos. The culture and the filtering methods we developed are easy to implement and require a very limited investment in equipment and consumables beside the standard one present in worm labs. In addition, this filtering method could be applied to nematode's species similar in size to C. elegans.


Maximum oxygen uptake and objectively measured physical activity in Danish children 6-7 years of age: the Copenhagen school child intervention study.
To provide normative data on maximum oxygen uptake (Vo(2)max) and physical activity in children 6-7 years of age and analyse the association between these variables. Vo(2)max was measured in 366 boys (mean (SD) 6.8 (0.4) years of age) and 332 girls (6.7 (0.4) years of age) from preschool classes in two suburban communities in Copenhagen, during a progressive treadmill exercise. Habitual physical activity was measured with accelerometers. Boys had higher Vo(2)max both in absolute values (1.19 (0.18) v 1.06 (0.16) litres/min (+11%), p<0.001) and relative to body weight (48.5 (6.0) v 44.8 (5.6) ml/kg/min (+8%); p<0.001) than girls. The difference in Vo(2)max between boys and girls decreased to +2% when expressed relative to lean body mass (LBM). Absolute Vo(2)max was related to LBM, body mass, and stature (all p<0.001). Boys were more physically active than girls (mean counts +9.4%, p<0.001), and even when boys and girls with the same Vo(2)max were compared, boys were more active. The difference in physical activity between the sexes was higher when sustained activity of higher intensity was compared. Vo(2)max is higher in boys than girls (+11%), even when related to body mass (+8%) and LBM (+2%). Most of the difference in Vo(2)max relative to body mass was explained by the larger percentage body fat in girls. When boys and girls with the same Vo(2)max were compared, boys engaged in more minutes of exercise of at least moderate intensity.


Pharmacotherapies for Adults With Alcohol Use Disorders: A Systematic Review and Network Meta-analysis.
We aimed to determine medications' comparative efficacy and safety for adults with alcohol use disorders. We searched eleven electronic data sources for randomized clinical trials with at least 4 weeks of treatment reporting on alcohol consumption (total abstinence and reduced heavy drinking), dropouts, and dropouts due to adverse events. We conducted network meta-analyses using random-effects, frequentist models, and calculated summary rate ratios (RRs) with 95% confidence intervals (CIs). We included 156 trials (N = 27,334). Nefazodone (RR = 2.11; 95% CI, 1.42-3.13), aripiprazole (RR = 1.97; 95% CI, 1.36-2.88), carbamazepine (RR = 1.85; 95% CI, 1.03-3.32), and nalmefene (RR = 1.17; 95% CI, 1.01-1.35) were associated with the most dropouts. Baclofen (RR = 0.83; 95% CI, 0.70-0.97) and pregabalin (RR = 0.63; 95% CI, 0.43-0.94) caused fewer dropouts than placebo. Nalmefene (RR = 3.26; 95% CI, 2.34-4.55), fluvoxamine (RR = 3.08; 95% CI, 1.59-5.94), and topiramate (RR=2.18; 95% CI, 1.36-3.51) caused more dropouts from adverse events over placebo. Gamma-hydroxy-butyrate (RR = 1.90; 95% CI, 1.03-3.53), baclofen (RR = 1.80; 95% CI, 1.39-2.34), disulfiram (RR = 1.71; 95% CI, 1.39-2.10), gabapentin (RR = 1.66; 95% CI, 1.04-2.67), acamprosate (RR = 1.33; 95% CI, 1.15-1.54), and oral naltrexone (RR = 1.15; 95% CI, 1.01-1.32) improved total abstinence over placebo (Fig. 3C). For reduced heavy drinking, disulfiram (RR = 0.19; 95% CI, 0.10-0.35), baclofen (RR = 0.72; 95% CI, 0.57-0.91), acamprosate (RR = 0.78; 95% CI, 0.70-0.86), and oral naltrexone (RR = 0.81; 95% CI, 0.73-0.90) were efficacious against placebo. The current meta-analyses provide evidence that several medications for AUDs are effective and safe and encourage the expanded use of these medications in the clinical setting. Our review found that acamprosate (2-3 g/d), disulfiram (250-500 mg/d), baclofen (30 mg/d), and oral naltrexone (50 mg/d) had the best evidence for improving abstinence and heavy drinking for patients with AUD. CRD42020208946.


A microcarrier cell culture system for large scale production of hepatitis A virus.
Hepatitis A virus (HAV) was isolated from human faeces using a fetal rhesus monkey kidney cell line (Frhk-4). Infectious medium from passage 12 was used to inoculate a large (5000 cm2) microcarrier cell culture maintained in suspension. The microcarriers used were swollen, collagen-coated dextran beads on which it was easy to propagate Frhk-4 cells. Intra- and extra-cellular virus levels were assayed and compared with conventional cultures in 25 cm2 plastic flasks. The results show that virus production per cell was similar in both systems. The number of cells per area unit in confluent cultures was initially lower in the microcarrier culture but subsequently increased. Two to three weeks post inoculation the virus yield per area unit in the microcarrier system was half of that of the conventional culture. The lower cell density per area unit in the microcarrier system was compensated by the large growth area that could be maintained in a single vessel and the total production of virus was substantial. Weekly harvests of medium with HAV antigen titres around 10(-2) contained antigenic material sufficient for several thousands of anti-HAV IgM tests. Propagation of HAV in microcarrier cell cultures thus seems a safe and simple way to produce large amounts of HAV.


Butyrate inhibits pro-proliferative miR-92a by diminishing c-Myc-induced miR-17-92a cluster transcription in human colon cancer cells.
Compromised colonic butyrate production resulting from low dietary fiber or altered gut microbiota may promote colon neoplasia. Previous reports indicate these actions are mediated in part by altered levels of miRNAs, including suppressed expression of the oncogenic miR-17-92a cluster. Here, we sought to identify the mechanisms underlying these effects of butyrate in colon cancer. miR-92a levels were measured in archived human colon cancer and adjacent normal colon specimens by microarray and quantitative RT-PCR (qPCR). The effects of butyrate and other histone deacetylase inhibitors (suberoylanilide hydroxamic acid (SAHA) and valproic acid) on primary (pri-miR17-92a), precursor and mature miR-92a were analyzed in HCT-116 and HT-29 human colon cancer cells using qPCR. The effects of butyrate, SAHA and valproic acid on protein levels of c-Myc, Drosha and p57 were measured in HCT-116 cells using immunoblotting. Regulation of C13orf25 promoter activity by butyrate was analyzed by luciferase reporter assay using modified pGL3 constructs containing a wild-type or mutated c-Myc binding site. Expression of c-Myc was modulated using siRNA or adenovirus vectors. p57 mRNA and protein were measured before and after transfection with miR-92a-mimic molecules. Following butyrate treatment and miR-92a-mimic transfection, apoptosis was analyzed by TUNEL staining and caspase-3 immunoblotting. Microarray, confirmed by qPCR, revealed a seven-fold increase in miR-92a levels in sporadic human colon cancer tissue compared to adjacent normal colon. Treating human colon cancer cells with butyrate reduced the levels of pri-miR17-92a, precursor and mature miR-92a, as well as c-Myc. SAHA and valproic acid had similar effects. Mutation of the c-Myc binding site diminished butyrate's inhibitory effects on C13orf25 promoter activity. Silencing c-Myc expression reduced miR-92a levels. c-Myc over-expression neutralized butyrate-induced attenuation of pri-miR17-92a. Exogenous miR-92a inhibited butyrate-induced p57 expression and reversed the beneficial actions of butyrate on colon cancer cell proliferation and apoptosis. Our findings identify a novel cellular mechanism whereby butyrate inhibits miR-92a transcription by reducing c-Myc, thus augmenting p57 levels. These actions diminish colon cancer cell proliferation and stimulate apoptosis. This newly described regulation of oncogenic miRNA biogenesis expands our understanding of colon cancer cell biology and identifies novel therapeutic targets.


Adaptive multi-player pursuit-evasion games with unknown general quadratic objectives.
In this paper, an adaptive parameter estimation scheme is investigated for a class of multi-player pursuit-evasion games subject to unknown objectives with linear feedback and cross terms in control strategy. In virtue of the adaptive method, each player updates controller strategies to estimate opponent's control parameters before getting the Nash equilibrium solution. Moreover, the sufficient condition is provided to guarantee stability of the proposed adaptive procedure. Meanwhile, the stability behavior of closed-loop Nash strategy with adaptive estimator is guaranteed by the contraction mapping method. Finally, a simulation of an all-against-one game with one escaper and three pursuers is carried out to demonstrate the effectiveness of the proposed methodology.


The effect of end screw orientation on the stability of anterior instrumentation in cyclic lateral bending.
Screw pullout at the proximal or distal end of multilevel anterior instrumentation can occur clinically. Previous laboratory studies have shown that angulation of vertebral body screws increases screw pullout strength and stability in toggling. To determine the effect of end screw angulation on instrumentation construct stability after cyclic, lateral bending. A biomechanical study in calf spines comparing two anterior spinal instrumentation constructs, one with parallel polyaxial screws and the other with angled polyaxial end screws. Sixteen instrumented constructs were made from eight thoracic (T8-T12) and eight lumbar calf spines (L1-L5). Eight (four lumbar specimens and four thoracic specimens) had five bicortical screws inserted mid-body and parallel to the end plates. The other eight specimens had two screws angled toward the superior end plates of the top two vertebrae; the middle vertebra had a mid-body screw parallel to the end plate, and the bottom two vertebrae had screws angled towards their inferior end plates. The constructs were then cycled in lateral bending, and the displacements of the two instrumentations with a 10 N-m bending load were compared. After 10,000 cycles, constructs with parallel end screws exhibited twice the average displacement than those with angled screws: 5.4 mm versus 2.9 mm (p=.031). The use of angled screws at the ends of anterior constructs demonstrated increased construct stability after cycling compared with traditional transverse screws. Although angled screw insertion is technically more difficult and is possible only with specific screw designs, its use might increase instrumentation longevity.