Intensity-Dependent Changes in Quantified Resting Cerebral Perfusion With Multiple Sessions of Transcranial DC Stimulation.
Transcranial direct current stimulation (tDCS) to the left prefrontal cortex has been shown to produce broad behavioral effects including enhanced learning and vigilance. Still, the neural mechanisms underlying such effects are not fully understood. Furthermore, the neural underpinnings of repeated stimulation remain understudied. In this work, we evaluated the effects of the repetition and intensity of tDCS on cerebral perfusion [cerebral blood flow (CBF)]. A cohort of 47 subjects was randomly assigned to one of the three groups. tDCS of 1- or 2-mA was applied to the left prefrontal cortex on three consecutive days, and resting CBF was quantified before and after stimulation using the arterial spin labeling MRI and then compared with a group that received sham stimulation. A widespread decreased CBF was found in a group receiving sham stimulation across the three post-stimulation measures when compared with baseline. In contrast, only slight decreases were observed in the group receiving 2-mA stimulation in the second and third post-stimulation measurements, but more prominent increased CBF was observed across several brain regions including the locus coeruleus (LC). The LC is an integral region in the production of norepinephrine and the noradrenergic system, and an increased norepinephrine/noradrenergic activity could explain the various behavioral findings from the anodal prefrontal tDCS. A decreased CBF was observed in the 1-mA group across the first two post-stimulation measurements, similar to the sham group. This decreased CBF was apparent in only a few small clusters in the third post-stimulation scan but was accompanied by an increased CBF, indicating that the neural effects of stimulation may persist for at least 24 h and that the repeated stimulation may produce cumulative effects.


Bacterial adherence to fibronectin and endothelial cells: a possible mechanism for bacterial tissue tropism.
In the pathophysiology of endocarditis, bacteria must initially adhere to the endothelial surface components of the cardiac valve before invasion and colonization. The attachment of bacteria to endothelial cell surfaces is dependent on surface characteristics of both the bacteria and the endothelial cell. Fibronectin, a glycoprotein produced by endothelial cells, binds to some bacteria but not all. This report examines bacterial adherence to human endothelial cells and to fibronectin-coated surfaces. Radiolabeled Staphylococcus aureus (Cowan I strain) and viridans streptococci avidly bound to fibronectin-coated surfaces and endothelial cell monolayers. In contrast, gram-negative organisms such as Escherichia coli bound poorly to both substrates. The adherence of S. aureus was dependent on time as well as on the concentration of fibronectin or the endothelial cell number. Bacterial adherence was specific for endothelial cells or fibronectin, since none occurred to plastic or to wells coated with collagen or albumin. The binding of S. aureus to fibronectin or endothelial cells appeared dependent on a bacterial surface protein, since treatment of the bacteria with proteases markedly decreased adherence. S. aureus was not dependent on the protein A or teichoic acid content of the cell wall, but adherence was markedly decreased in bacterial strains that had a carbohydrate capsule. S. aureus pretreated with serum or purified fibronectin manifested enhanced adherence to endothelial cells, suggesting fibronectin-fibronectin interactions. Bacteria specifically attach to endothelial cells and to fibronectin-coated surfaces, which suggests that the ability of a bacterium to attach to these substrates may reflect the propensity to invade and colonize vascular tissues such as cardiac valves.


Pulmonary infections in liver transplant recipients receiving tacrolimus. Changing pattern of microbial etiologies.
Pulmonary infections are a significant cause of morbidity after liver transplantation; Gram-negative bacilli, cytomegalovirus, and Pneumocystis carinii were the usual pulmonary pathogens in the earlier studies in liver transplant recipients receiving cyclosporine. We prospectively assessed the impact of pulmonary infection in 101 consecutive liver transplant recipients receiving the new immunosuppressive agent tacrolimus (FK506). Fifteen percent (15/101) of the patients had 19 episodes of pneumonia; 58% (11/19) of the pneumonias were bacterial, 37% (7/19) were fungal, and 5% (1/19) were protozoal (Toxoplasma gondii). Twenty-seven percent of the bacterial pneumonias were due to Legionella. None of the patients had cytomegalovirus or P carinii pneumonia. Seven percent (7/10) of the study patients had fungal pneumonitis; 4% had invasive aspergillosis and 3% had cryptococcosis. Mortality was significantly higher (53%, 8/15) for patients with pneumonia than for patients without pneumonia (10%, 9/86, P = 0.0004). Only fungal pneumonias were the direct cause of death; 63% (5/8) of the deaths were in patients with fungal pneumonitis. Our data suggest a changing pattern of microbial etiologies of pneumonitis in the era of modern immunosuppressive agents. We show that P carinii pneumonia and cytomegalovirus can be effectively curtailed with appropriate prophylaxis. Fungal infections, on the contrary, not only constituted a major proportion of the pneumonia, but also carried the highest pneumonia-associated mortality. Legionella infections can be overlooked unless specialized laboratory methodology (cultured on selective media, urinary antigen) are applied routinely on all cases of pneumonia. We recommend routine culture on the water supply for Legionella in all transplant centers.


Variation in outpatient oral antimicrobial use patterns among Canadian provinces, 2000 to 2010.
The volume and patterns of antimicrobial drug use are key variables to consider when developing guidelines for prescribing, and programs to address stewardship and combat the increasing prevalence of antimicrobial resistant pathogens. Because drug programs are regulated at the provincial level, there is an expectation that antibiotic use may vary among provinces. To assess these potential differences according to province and time. Provincial antimicrobial prescribing data at the individual drug level were acquired from the Canadian Integrated Program for Antimicrobial Resistance Surveillance for 2000 to 2010. Data were used to calculate two yearly metrics: prescriptions per 1000 inhabitant-days and the average defined daily doses per prescription. The proportion of liquid oral prescriptions of total prescriptions was also calculated as a proxy measure for the proportion of prescriptions given to children versus adults. To assess the significance of provincial antimicrobial use, linear mixed models were developed for each metric, accounting for repeated measurements over time. Significant differences among provinces were found, as well as significant changes in use over time. Newfoundland and Labrador was found to have significantly higher prescribing rates than all other provinces (P<0.001) in 2010, as well as the mean of all other provinces (P<0.001). In contrast, Quebec exhibited significantly lower prescribing than all other provinces (P<0.001 for all provinces except British Columbia, where P=0.024) and the mean of all other provinces (P<0.001). Reports of reductions in antimicrobial use at the Canadian level are promising, especially prescribing to children; however, care must be taken to avoid the pitfall of the ecological fallacy. Reductions are not consistent among the provinces or among the classes of antimicrobial drugs dispensed in Canada.


LncRNA PSMB8-AS1 Instigates Vascular Inflammation to Aggravate Atherosclerosis.
Increasing evidence suggests that long noncoding RNAs play significant roles in vascular biology and disease development. One such long noncoding RNA, PSMB8-AS1, has been implicated in the development of tumors. Nevertheless, the precise role of PSMB8-AS1 in cardiovascular diseases, particularly atherosclerosis, has not been thoroughly elucidated. Thus, the primary aim of this investigation is to assess the influence of PSMB8-AS1 on vascular inflammation and the initiation of atherosclerosis. We generated PSMB8-AS1 knockin and Apoe (Apolipoprotein E) knockout mice (Apoe-/-PSMB8-AS1KI) and global Apoe and proteasome subunit-β type-9 (Psmb9) double knockout mice (Apoe-/-Psmb9-/-). To explore the roles of PSMB8-AS1 and Psmb9 in atherosclerosis, we fed the mice with a Western diet for 12 weeks. Long noncoding RNA PSMB8-AS1 is significantly elevated in human atherosclerotic plaques. Strikingly, Apoe-/-PSMB8-AS1KI mice exhibited increased atherosclerosis development, plaque vulnerability, and vascular inflammation compared with Apoe-/- mice. Moreover, the levels of VCAM1 (vascular adhesion molecule 1) and ICAM1 (intracellular adhesion molecule 1) were significantly upregulated in atherosclerotic lesions and serum of Apoe-/-PSMB8-AS1KI mice. Consistently, in vitro gain- and loss-of-function studies demonstrated that PSMB8-AS1 induced monocyte/macrophage adhesion to endothelial cells and increased VCAM1 and ICAM1 levels in a PSMB9-dependent manner. Mechanistic studies revealed that PSMB8-AS1 induced PSMB9 transcription by recruiting the transcription factor NONO (non-POU domain-containing octamer-binding protein) and binding to the PSMB9 promoter. PSMB9 (proteasome subunit-β type-9) elevated VCAM1 and ICAM1 expression via the upregulation of ZEB1 (zinc finger E-box-binding homeobox 1). Psmb9 deficiency decreased atherosclerotic lesion size, plaque vulnerability, and vascular inflammation in Apoe-/- mice in vivo. Importantly, endothelial overexpression of PSMB8-AS1-increased atherosclerosis and vascular inflammation were attenuated by Psmb9 knockout. PSMB8-AS1 promotes vascular inflammation and atherosclerosis via the NONO/PSMB9/ZEB1 axis. Our findings support the development of new long noncoding RNA-based strategies to counteract atherosclerotic cardiovascular disease.


[Growth factors as neuroprotective treatment in Parkinson disease?].
One of several probable causation theories of Parkinson disease postulates that brain tissue cannot generate sufficient levels of various growth factors required to sustain the viability of dopamine-producing nerve cells in the presence of as yet unknown toxic factors. The study reported here evaluates the ability of externally applied growth factors to protect the dopamine fibres in the basal ganglia in a toxin-induced animal model of the disease. All animals (rats) were subjected to selective destruction of the dopamine-producing cells in substantia nigra. The rats were divided into three groups. Two groups received intracerebral treatment with either glia-cell derived neurotrophic factor (GDNF) or a combination of brain-derived neurotrophic factor (BDNF) and GDNF. The third group acted as untreated controls and were given sterile saline. The growth factors were infused directly into the brain by an osmotic pump over a period of 28 days. Brain sections taken from all three groups were evaluated by immunocytochemistry. The two groups of rats that received growth factor infusion displayed a significant improvement in their motor behaviour compared to control animals. Immunocytochemistry studies demonstrated that the group receiving a combination of GDNF and BDNF had an increased number of surviving active fibres in the dopamine system striatum in comparison to the control and GDNF groups. In addition the infusion of growth factors resulted in a proliferation of subventricular cells in the basal ganglia. The improved motor function following growth factor treatment in this rat model might be due to a delayed retrograde degeneration of the nigrostriatal nerve fibers. Growth factor infusion also clearly stimulated endogenous stem cells and caused their migration towards the striatum. Our observations indicate that the infusion of growth factors into the brain have a symptomatic and neuroprotective effect in this model.


Weight, inflammation, cancer-related symptoms and health related quality of life among breast cancer survivors.
Maintaining weight is important for better prognosis of breast cancer survivors. The associations between weight and cancer-related symptoms are not known. We examined associations among weight, weight change, inflammation, cancer-related symptoms, and health-related quality of life (HRQOL) in a cohort of stage 0-IIIA breast cancer survivors. Participants were recruited on average 6 months (2–12 months) after diagnosis. Height, weight, and C-reactive protein (CRP) were assessed at approximately 30 months post-diagnosis; cancer-related symptoms (chest wall and arm symptoms, vasomotor symptoms, urinary incontinence, vaginal symptoms, cognition/mood problems, sleep, sexual interest/function), and HRQOL (SF-36) were assessed at approximately 40 months post-diagnosis. Weight was measured at baseline in a subset. Data on 661 participants were evaluable for body mass index (BMI); 483 were evaluable for weight change. We assessed associations between BMI (<25.0, 25.0–29.9, ≥30.0 kg/m2), post-diagnosis weight change (lost ≥5 %, weight change <5 %, gained ≥5 %), and CRP (tertile) with cancer-related symptoms and HRQOL using analysis of covariance. Higher symptoms scores indicate more frequent or severe symptoms. Higher HRQOL scores indicate better HRQOL. Compared with those with BMI <25 kg/m2, women with BMI ≥30 kg/m2 had the following scores: increased for arm symptoms (+25.0 %), urinary incontinence (+40.0 %), tendency to nap (+18.9 %), and poorer physical functioning (−15.6 %, all p < 0.05). Obese women had lower scores in trouble falling asleep (−9.9 %; p < 0.05). Compared with weight change <5 %, participants with ≥5 % weight gain had lower scores in physical functioning (−7.2 %), role-physical (−15.5 %) and vitality (−11.2 %), and those with weight loss ≥5 % had lower chest wall (−33.0 %) and arm symptom scores (−35.5 %, all p < 0.05). Increasing CRP tertile was associated with worse scores for chest wall symptoms, urinary incontinence, physical functioning, role-physical, vitality and physical component summary scores (all P trend < 0.05). Future studies should examine whether interventions to maintain a healthy weight and reduce inflammation could alleviate cancer-related symptoms and improve HRQOL.


Nanoscale Membrane Domain Formation Driven by Cholesterol.
Biological membranes generate specific functions through compartmentalized regions such as cholesterol-enriched membrane nanodomains that host selected proteins. Despite the biological significance of nanodomains, details on their structure remain elusive. They cannot be observed via microscopic experimental techniques due to their small size, yet there is also a lack of atomistic simulation models able to describe spontaneous nanodomain formation in sufficiently simple but biologically relevant complex membranes. Here we use atomistic simulations to consider a binary mixture of saturated dipalmitoylphosphatidylcholine and cholesterol - the "minimal standard" for nanodomain formation. The simulations reveal how cholesterol drives the formation of fluid cholesterol-rich nanodomains hosting hexagonally packed cholesterol-poor lipid nanoclusters, both of which show registration between the membrane leaflets. The complex nanodomain substructure forms when cholesterol positions itself in the domain boundary region. Here cholesterol can also readily flip-flop across the membrane. Most importantly, replacing cholesterol with a sterol characterized by a less asymmetric ring region impairs the emergence of nanodomains. The model considered explains a plethora of controversial experimental results and provides an excellent basis for further computational studies on nanodomains. Furthermore, the results highlight the role of cholesterol as a key player in the modulation of nanodomains for membrane protein function.


Virulence gene regulation by CvfA, a putative RNase: the CvfA-enolase complex in Streptococcus pyogenes links nutritional stress, growth-phase control, and virulence gene expression.
Streptococcus pyogenes, a multiple-auxotrophic human pathogen, regulates virulence gene expression according to nutritional availability during various stages in the infection process or in different infection sites. We discovered that CvfA influenced the expression of virulence genes according to growth phase and nutritional status. The influence of CvfA in C medium, rich in peptides and poor in carbohydrates, was most pronounced at the stationary phase. Under these conditions, up to 30% of the transcriptome exhibited altered expression; the levels of expression of multiple virulence genes were altered, including the genes encoding streptokinase, CAMP factor, streptolysin O, M protein (more abundant in the CvfA(-) mutant), SpeB, mitogenic factor, and streptolysin S (less abundant). The increase of carbohydrates or peptides in media restored the levels of expression of the virulence genes in the CvfA(-) mutant to wild-type levels (emm, ska, and cfa by carbohydrates; speB by peptides). Even though the regulation of gene expression dependent on nutritional stress is commonly linked to the stringent response, the levels of ppGpp were not altered by deletion of cvfA. Instead, CvfA interacted with enolase, implying that CvfA, a putative RNase, controls the transcript decay rates of virulence factors or their regulators according to nutritional status. The virulence of CvfA(-) mutants was highly attenuated in murine models, indicating that CvfA-mediated gene regulation is necessary for the pathogenesis of S. pyogenes. Taken together, the CvfA-enolase complex in S. pyogenes is involved in the regulation of virulence gene expression by controlling RNA degradation according to nutritional stress.


Responsiveness to acidity via metal ion regulators mediates virulence in the gastric pathogen Helicobacter pylori.
The virulence of pathogenic bacteria is dependent on their adaptation to and survival in the stressful conditions encountered in their hosts. Helicobacter pylori exclusively colonizes the acid stomach of primates, making it an ideal study model. Little is known about how H. pylori responds to the moderately acidic conditions encountered at its colonization site, the gastric mucus layer. Thus, we compared gene expression profiles of H. pylori 26695 grown at neutral and acidic pH, and validated the data for a selection of genes by real-time polymerase chain reaction, dot-blots or enzymatic assays. During growth in acidic conditions, 56 genes were upregulated and 45 genes downregulated. We found that acidity is a signal modulating the expression of several virulence factors. Regulation of genes related to metal ion homeostasis suggests protective mechanisms involving diminished transport and enhanced storage. Genes encoding subunits of the F0F1 ATPase and of a newly identified Na+/H+ antiporter (NhaC-HP0946) were downregulated, revealing that this bacterium uses original mechanisms to control proton entry. Five of the upregulated genes encoded proteins controlling intracellular ammonia synthesis, including urease, amidase and formamidase, underlining the major role of this buffering compound in the protection against acidity in H. pylori. Regulatory networks and transcriptome analysis as well as enzymatic assays implicated two metal-responsive transcriptional regulators (NikR and Fur) and an essential two-component response regulator (HP0166, OmpR-like) as effectors of the H. pylori acid response. Finally, a nikR-fur mutant is attenuated in the mouse model, emphasizing the link between response to acidity, metal metabolism and virulence in this gastric pathogen.


The effects of surface vegetation coverage on the spatial distribution of PM2.5 in the central area of Nanchang City, China.
The frequent occurrence of haze has caused widespread concern in China, and PM2.5 is thought to be the main cause. Previous research showed that PM2.5 was not only influenced by meteorological conditions but also by land cover especially surface vegetation. It was concluded that PM2.5 concentration is significantly influenced by surface vegetation, but spatially how and in what manner are still unanswered. Taking the central area of Nanchang City, China, as a case, this study firstly applied land use regression (LUR) model to simulate the distribution of PM2.5 in 2020. Then, the dichotomous model was used to determine vegetation coverage. A statistical regression model was used to analyze the influence of vegetation cover on PM2.5 and the scale effects. The results showed that (1) vegetation coverage and PM2.5 concentration were both significantly negatively correlated at the spatial scales selected for this study. (2) The effect of vegetation coverage on PM2.5 varied with season and the 500 m had the best correlation. (3) The non-linear regression model fits better than the linear model, and the effect of vegetation coverage on PM2.5 was complex. (4) The effect of vegetation coverage on PM2.5 concentration was different with PM2.5 concentration level. The higher the PM2.5 concentration, the more pronounced the effect of vegetation coverage on it. This study proposed the idea and method of coupling vegetation coverage with PM2.5 concentration at the regional scale from gradient landscape's point of view and provided some references for mitigating PM2.5 pollution through optimizing urban vegetation patterns.


Enzymatic and Chemoenzymatic Three-Step Cascades for the Synthesis of Stereochemically Complementary Trisubstituted Tetrahydroisoquinolines.
Chemoenzymatic and enzymatic cascade reactions enable the synthesis of complex stereocomplementary 1,3,4-trisubstituted tetrahydroisoquinolines (THIQs) with three chiral centers in a step-efficient and selective manner without intermediate purification. The cascade employs inexpensive substrates (3-hydroxybenzaldehyde and pyruvate), and involves a carboligation step, a subsequent transamination, and finally a Pictet-Spengler reaction with a carbonyl cosubstrate. Appropriate selection of the carboligase and transaminase enzymes enabled the biocatalytic formation of (1R,2S)-metaraminol. Subsequent cyclization catalyzed either enzymatically by a norcoclaurine synthase or chemically by phosphate resulted in opposite stereoselectivities in the products at the C1 position, thus providing access to both orientations of the THIQ C1 substituent. This highlights the importance of selecting from both chemo- and biocatalysts for optimal results.


[The latent structure of psychodynamic and psychological personality models: Conjoint investigation of OPD-CQ, OPD-SQS and BFI-2].
The latent structure of psychodynamic and psychological personality models: Conjoint investigation of OPD-CQ, OPD-SQS and BFI-2 Objectives: Meanwhile, psychodynamic self-report questionnaires for the assessment of structure and conflict as aspects of personality have been developed. References to the psychological Big Five model, which is becoming more important due to the ongoing paradigm shift in the medical classification systems of personality disorders, are still unclear. Methods: The hierarchical factor structure of the psychodynamic scales of the OPD Conflict Questionnaire (OPD-CQ) and the short version of the OPD Structure Questionnaire (OPDSQS) as well as the psychological scales of the Big Five Inventory 2 (BFI-2) was investigated. The study was based on a representative sample of the German working population considering gender, age and place of residence (N = 545). Results: Factor analysis suggested the seven factors Negative Emotionality, Extraversion, Structural Impairment, Conscientiousness, Openness, Dissociality and Oedipal Conflict (explained variance = 56.84 %). Overlaps between psychodynamic and psychological scales were found with regard to Negative Emotionality, Extraversion and Agreeableness. Psychodynamic scales did not cover Conscientiousness and Openness, while the level of personality functioning and the role of sexuality were not covered by psychological scales. Discussion: The study provides important information regarding the relationship between psychodynamic concepts and the Big Five.


Anti-basal ganglia antibodies in PANDAS.
An autoimmune-mediated mechanism involving molecular mimicry has been proposed for a variety of pediatric movement disorders that occur after a streptococcal infection. In this study, anti-basal ganglia antibodies (ABGA) were measured in 15 children with the diagnosis of pediatric autoimmune neuropsychiatric disorder associated with streptococcal infection (PANDAS) and compared with those in 15 controls. ELISA and Western immunoblotting (WB) methods were used to detect ABGA against supernatant (S1), pellet (P2), and synaptosomal preparations from adult postmortem caudate, putamen, and globus pallidus. ELISA optical density values did not differ between PANDAS patients and controls across all preparations. Immunoblotting identified multiple bands in all subjects with no differences in the number of bands or their total density. Discriminant analysis, used to assess mean binding patterns, showed that PANDAS patients differed from controls only for the caudate S1 fraction (Wilks' lambda = 0.0236, P < 0.0002), with PANDAS-primarily tic subjects providing the greatest discrimination. Among the epitopes contributing to differences between PANDAS and control in the caudate S1 fraction, mean binding to the epitope at 183 kDa was the most different between groups. In conclusion, ELISA measurements do not differentiate between PANDAS and controls, suggesting a lack of major antibody changes in this disorder. Further immunoblot analyses using a caudate supernatant fraction are required to completely exclude the possibility of minor antibody repertoire differences in PANDAS subjects, especially in those who primarily have tics.


The interplay between determinism and stochasticity in childhood diseases.
An important issue in the history of ecology has been the study of the relative importance of deterministic forces and processes noise in shaping the dynamics of ecological populations. We address this question by exploring the temporal dynamics of two childhood infections, measles and whooping cough, in England and Wales. We demonstrate that epidemics of whooping cough are strongly influenced by stochasticity; fully deterministic approaches cannot achieve even a qualitative fit to the observed data. In contrast, measles dynamics are extremely well explained by a deterministic model. These differences are shown to be caused by their contrasting responses to dynamical noise due to different infectious periods.


Effect of liver injury on prognosis and treatment of hospitalized patients with COVID-19 pneumonia.
Liver injury is common in patients with coronavirus disease 2019 (COVID-19), although its effect on patient outcomes has not been well studied. This study aimed to evaluate the effect of liver injury on the prognosis and treatment of patients with COVID-19 pneumonia. In this retrospective, single-center study, data on 109 hospitalized patients with COVID-19 pneumonia were extracted and analyzed. The primary composite end-point event was the use of mechanical ventilation or death. At admission, of the 109 patients enrolled, 56 patients (51.4%) were diagnosed with severe disease, and 39 (35.8%) presented with liver injury, which mainly manifested as elevated levels of alanine aminotransferase (ALT) or aspartate aminotransferase (AST) accompanied simultaneously by an increase in the level of γ-glutamyl transferase. A primary composite end-point event occurred in 21 patients (19.3%). Liver injury was more prevalent in patients with severe disease than in those with non-severe disease (46.4% vs. 24.5%, P=0.017). However, there was no significant difference found between severe and non-severe patients in the use of mechanical ventilation, mortality, hospital stay, or use and dosage of glucocorticoids between individuals with and without liver injury (all P>0.05). The degree of disease severity (OR =7.833, 95% CI, 1.834-31.212, P=0.005) and presence of any coexisting illness (OR =4.736, 95% CI, 1.305-17.186, P=0.018) were predictable risk factors for primary composite end-point events, whereas liver injury had no significance in this aspect (OR =0.549, 95% CI, 0.477-5.156, P=0.459). Liver injury was more common in severe cases of COVID-19 pneumonia than in non-severe cases. However, liver injury had no negative effect on the prognosis and treatment of COVID-19 pneumonia.


Participation of two members of the very long-chain acyl-CoA synthetase family in bile acid synthesis and recycling.
Bile acids are synthesized de novo in the liver from cholesterol and conjugated to glycine or taurine via a complex series of reactions involving multiple organelles. Bile acids secreted into the small intestine are efficiently reabsorbed and reutilized. Activation by thioesterification to CoA is required at two points in bile acid metabolism. First, 3alpha,7alpha,12alpha-trihydroxy-5beta-cholestanoic acid, the 27-carbon precursor of cholic acid, must be activated to its CoA derivative before side chain cleavage via peroxisomal beta-oxidation. Second, reutilization of cholate and other C24 bile acids requires reactivation prior to re-conjugation. We reported previously that homolog 2 of very long-chain acyl-CoA synthetase (VLCS) can activate cholate (Steinberg, S. J., Mihalik, S. J., Kim, D. G., Cuebas, D. A., and Watkins, P. A. (2000) J. Biol. Chem. 275, 15605-15608). We now show that this enzyme also activates chenodeoxycholate, the secondary bile acids deoxycholate and lithocholate, and 3alpha,7alpha,12alpha-trihydroxy-5beta-cholestanoic acid. In contrast, VLCS activated 3alpha,7alpha,12alpha-trihydroxy-5beta-cholestanoate, but did not utilize any of the C24 bile acids as substrates. We hypothesize that the primary function of homolog 2 is in the reactivation and recycling of C24 bile acids, whereas VLCS participates in the de novo synthesis pathway. Results of in situ hybridization, topographic orientation, and inhibition studies are consistent with the proposed roles of these enzymes in bile acid metabolism.


Gallionellaceae in rice root plaque: metabolic roles in iron oxidation, nutrient cycling, and plant interactions.
In waterlogged soils, iron plaque forms a reactive barrier between the root and soil, collecting phosphate and metals such as arsenic and cadmium. It is well established that iron-reducing bacteria solubilize iron, releasing these associated elements. In contrast, microbial roles in plaque formation have not been clear. Here, we show that there is a substantial population of iron oxidizers in plaque, and furthermore, that these organisms (Sideroxydans and Gallionella) are distinguished by genes for plant colonization and nutrient fixation. Our results suggest that iron-oxidizing and iron-reducing bacteria form and remodel iron plaque, making it a dynamic system that represents both a temporary sink for elements (P, As, Cd, C, etc.) as well as a source. In contrast to abiotic iron oxidation, microbial iron oxidation results in coupled Fe-C-N cycling, as well as microbe-microbe and microbe-plant ecological interactions that need to be considered in soil biogeochemistry, ecosystem dynamics, and crop management.


Translabyrinthine removal of large acoustic tumors.
The translabyrinthine technique was used to remove tumors of various sizes in 50 cases. Outcome is presented stressing immediate and eventual facial function, for tumors measuring 20, 25, 30, 35, and 40 mm. Immediate facial function is a good predictor of permanent facial activity; regeneration of the facial nerve gives good but never perfect function. Except for preservation of hearing, translabyrinthine surgery gives equally good and probably better results than other techniques.


[Neonatal lupus in an infant of a mother followed up for dermatomyositis: medical images].
Neonatal lupus is rare. It is due to the transmission of maternal autoantibodies across the placenta during pregnancy. We here report the case of a 2-month old female infant treated for erythematous macular cutaneous lesions on the face and the trunk. Her mother was followed up for dermatomyositis diagnosed on the basis of clinical lesions, muscle weakness and elevation in muscle enzyme levels. However she had not underwent antinuclear antibody test (ANA). Clinical examination showed atrophic erythematous lesions distributed like butterfly wings on both sides of the nasal pyramid, satellite lesions on the front, with red hair. The remainder of the physical examination was unremarkable. Laboratory tests were not performed because infant's parents also refused the biopsy. These lesions suggested seborrheic dermatitis, rosacea or atopic dermatitis. However, rosacea is very rare in infants and usually affects fair-skinned people. In seborrheic dermatitis, lesions are not atrophic. The age of onset of atopic dermatitis is usually 3 months. Lesions regressed in 15 days under dermocorticoid therapy.


Biphasic survival analysis of trypanotolerance QTL in mice.
A marker-assisted introgression (MAI) experiment was conducted to transfer trypanotolerance quantitative trait loci (QTL) from a donor mouse strain, C57BL/6, into a recipient mouse strain, A/J. The objective was to assess the effect of three previously identified chromosomal regions on mouse chromosomes 1 (MMU1), 5 (MMU5) and 17 (MMU17) in different genetic backgrounds on the survival pattern following infection with Trypanosoma congolense. An exploratory data analysis revealed a biphasic pattern of time to death, with highly distinct early and late mortality phases. In this paper, we present survival analysis methods that account for the biphasic mortality pattern and results of reanalyzing the data from the MAI experiment. The analysis with a Weibull mixture model confirmed the biphasic pattern of time to death. Mortality phase, an unobserved variable, appears to be an important factor influencing survival time and is modeled as a binary outcome variable using logistic regression analysis. Accounting for this biphasic pattern in the analysis reveals that a previously observed sex effect on average survival is rather an effect on proportion of mice in the two mortality phases. The C57BL/6 (donor) QTL alleles on MMU1 and MMU17 act dominantly in the late mortality phase while the A/J (recipient) QTL allele on MMU17 acts dominantly in the early mortality phase. From this study, we found clear evidence for a biphasic survival pattern and provided models for its analysis. These models can also be used when studying defense mechanisms against other pathogens. Finally, these approaches provide further information on the nature of gene actions.


Chitosan/siRNA functionalized titanium surface via a layer-by-layer approach for in vitro sustained gene silencing and osteogenic promotion.
Titanium surface modification is crucial to improving its bioactivity, mainly its bone binding ability in bone implant materials. In order to functionalize titanium with small interfering RNA (siRNA) for sustained gene silencing in nearby cells, the layer-by-layer (LbL) approach was applied using sodium hyaluronate and chitosan/siRNA (CS/siRNA) nanoparticles as polyanion and polycation, respectively, to build up the multilayered film on smooth titanium surfaces. The CS/siRNA nanoparticle characterization was analyzed first. Dynamic contact angle, atomic force microscopy, and scanning electron microscopy were used to monitor the layer accumulation. siRNA loaded in the film was quantitated and the release profile of film in phosphate-buffered saline was studied. In vitro knockdown effect and cytotoxicity evaluation of the film were investigated using H1299 human lung carcinoma cells expressing green fluorescent protein (GFP). The transfection of human osteoblast-like cell MG63 and H1299 were performed and the osteogenic differentiation of MG63 on LbL film was analyzed. The CS/siRNA nanoparticles exhibited nice size distribution. During formation of the film, the surface wettability, topography, and roughness were alternately changed, indicating successful adsorption of the individual layers. The scanning electron microscope images clearly demonstrated the hybrid structure between CS/siRNA nanoparticles and sodium hyaluronate polymer. The cumulated load of siRNA increased linearly with the bilayer number and, more importantly, a gradual release of the film allowed the siRNA to be maintained on the titanium surface over approximately 1 week. In vitro transfection revealed that the LbL film-associated siRNA could consistently suppress GFP expression in H1299 without showing significant cytotoxicity. The LbL film loading with osteogenic siRNA could dramatically increase the osteogenic differentiation in MG63. In conclusion, LbL technology can potentially modify titanium surfaces with specific gene-regulatory siRNAs to enhance biofunction.


Effects of sedation, anesthesia, and endotracheal intubation on respiratory mechanics in adult horses.
To determine the effects of endotracheal intubation on respiratory mechanics during xylazine sedation and xylazine-diazepam-ketamine anesthesia in adult horses. 5 healthy adult horses. Measurements were derived from recordings of respiratory gas flow, and transpulmonary and transtracheal pressures. Total pulmonary resistance (RT) was partitioned into upper airway resistance (extrathoracic portion of trachea, larynx, pharynx, nasal cavity, nares; RUA) and lower airway resistance (intrathoracic portion of trachea, bronchi, bronchioles). Baseline measurements were obtained in unsedated horses, after xylazine administration, and following nasotracheal intubation (ID, 18 mm). Measurements were obtained following induction of xylazine-diazepam-ketamine anesthesia and subsequent to endotracheal intubations (ID, 22, 20, and 16 mm). During recovery, horses were nasotracheally intubated (ID, 18 mm). Measurements were obtained upon standing, and repeated after extubation. Data were examined by use of ANOVA with repeated measures. Significant increases in mean work of breathing (W), RT, and RUA observed with xylazine sedation were variably attenuated by nasotracheal intubation. During xylazine-diazepam-ketamine anesthesia, the highest mean values for W, RT, RUA, transpulmonary and transtracheal pressures developed during non-intubation periods. The magnitudes of resistance and pressure values were inversely proportional to the internal diameter of the endotracheal tube. At recovery, values of the W and all measurements of resistances and pressures were significantly increased, compared with presedation values. Extubation resulted in further increases in these measurements. Work of breathing in horses is substantially increased when RUA is increased during xylazine sedation and xylazine-diazepam-ketamine anesthesia. Endotracheal intubation reduces W by reducing RUA.


An analysis of selected factors that affect the work lives of athletic trainers employed in accredited educational programs.
To gain insight into selected factors that affect certified athletic trainers who work in accredited athletic train- ing educational programs. Specifically, the demands and ex- pectations placed upon certified athletic trainers relative to workloads, elements of compensation such as salary and benefits, and performance evaluation criteria were studied. We designed the Survey of Athletic Trainer Workloads, Criteria for Retention and Promotion, and Salary Levels. Three copies were sent to all undergraduate curriculum directors for accredited athletic training education programs (n = 88). One hundred fifty-three usable surveys were returned (58%). The subjects were 153 certified athletic trainers who were employed in accredited athletic training education programs. Simple descriptive data including frequency distributions and measures of central tendency were used to compile most of the results and findings. A 2-way analysis of variance was performed relating to salary, years of experience, and sex. Most certified athletic trainers (78%) worked on staffs of at least 3 certified athletic trainers. Almost universally, responsibilities included some combination of teaching, athletic service, and student supervision. Certified athletic trainers reported being certified an average of 12.5 years and typically worked more than 50 hours per week. The greatest percentage of those responding (56%) earned between $25,000 and $39,999 annually. Salaries were significantly lower for females than for their male counterparts of equal experience (P < .05). Chairperson evaluations and student evaluations on teaching were rated as the most important performance evaluation criteria for athletic trainers regarding merit and promotion. Although there was a wide range of responses for most categories surveyed, the data revealed clear trends. With more information of this nature, realistic assessments and comparisons can be made among certified athletic trainers working in accredited curriculum programs.


Pregnancy-associated glycoproteins in cows with retained fetal membranes.
In cows, retained fetal membranes (RFM) are a major problem in reproduction. The timely detachment of fetal membranes after parturition requires well coordinated maturation processes in the placenta. One feature of placental maturation in cows is a prepartal decline in the number of binucleate trophoblast giant cells (BNC) in the fetal chorion. Pregnancy associated glycoproteins (PAGs) are a group of proteins, produced by trophoblast cells in artiodactyls. We studied aspects of PAG expression in cows with and without RFM. The numerical density of PAG-positive immunostained BNC in placentomal samples, collected from cows with normal expulsion of fetal membranes (n = 20) and cows with RFM (n = 20) was determined. The number of PAG-positive BNCs was significantly higher in cows with RFM, compared to controls. The concentration of PAGs in maternal serum in prepartum, intrapartum, and postpartum cows was measured (RFM n = 20; controls n = 68). No significant differences between RFM and controls were detected. Microarray analysis of placental PAG mRNA expression was done with two types of microarrays: Affymetrix (RFM n = 20; controls n = 20) and Agilent (RFM n = 8; controls n = 8). Both microarrays showed a significantly higher expression of modern PAGs in RFM cases. Our results show that the expression of modern PAGs, which are produced by BNCs and are secreted into the maternal organism, are differentially expressed in RFM. Although the concentration in peripheral maternal blood did not differ between RFM and controls, the local concentration in the placenta is likely to be higher in RFM cases. This suggests the possibility of local regulatory roles of PAG in the release of bovine fetal membranes.


Impaired heart rate variability in euthymic bipolar patients.
Power spectral analysis of heart rate variability (HRV) has recently been shown to be a reliable non-invasive test for quantitative assessment of the central sympathovagal interaction that modulates cardiovascular autonomic function. We studied 39 euthymic bipolar patients and 39 controls, matched for age and sex. A high-resolution electrocardiogram was obtained during complete rest. Spectral analysis of R-R intervals was performed by the fast Fourier transform algorithm. Euthymic bipolar patients at rest are characterized by markedly low HRV, independent of specific drug treatments. While these results of autonomic stability in euthymic bipolar patients are counter-intuitive, the findings may be important in understanding pathophysiology of bipolar affective illness.


Exploiting apoptosis for therapeutic tolerance induction.
Immune tolerance remains the most promising yet elusive strategy for treating immune-mediated diseases. An experimental strategy showing promise in phase 1 clinical studies is the delivery of Ag cross-linked to apoptotic leukocytes using ethylene carbodiimide. This approach originated from demonstration of the profound tolerance-inducing ability of i.v. administered Ag-coupled splenocytes (Ag-SP) in mice, which has been demonstrated to treat T cell-mediated disorders including autoimmunity, allergy, and transplant rejection. Recent studies have defined the intricate interplay between the innate and adaptive immune systems in Ag-SP tolerance induction. Innate mechanisms include scavenger receptor-mediated uptake of Ag-SP by host APCs, Ag representation, and the required upregulation of PD-L1 expression and IL-10 production by splenic marginal zone macrophages leading to Ag-specific T cell regulation via the combined effects of cell-intrinsic anergy and regulatory T cell induction. In this paper, we discuss the history, advantages, current mechanistic understanding, and clinical potential of tolerance induction using apoptotic Ag-coupled apoptotic leukocytes.


Masculinity, infertility, stigma and media reports.
There is growing concern about the health of men in the developed West. Compared with women they have higher rates of morbidity and mortality and are less likely to seek out and employ medical services. Several authors have drawn on social constructionist models, such as the concept of hegemonic masculinity, to account for these gender differences in risk and behaviour. One might anticipate that certain conditions, such as male infertility, would be perceived as posing a particular threat to conventional views of masculinity. There is some support for this, although there is little research into the social construction of male infertility. In this study Discourse Analysis was employed to analyse newspaper accounts of a reported decline in sperm counts in order to study the way in which infertility and masculinity were represented and constructed in the media. The results indicate a construction of fertility as being in crisis and of male infertility as conflated with impotence. Men were positioned as vulnerable and threatened by forces outside their control. The accounts drew on a range of stereotypically masculine reference points, such as warfare and mechanical analogies. These results are consistent with concepts of hegemonic masculinity and suggest that men are offered a highly restricted set of options in terms of perceiving and representing their bodies and their health.


Tmem100 Is a Regulator of TRPA1-TRPV1 Complex and Contributes to Persistent Pain.
TRPA1 and TRPV1 are crucial pain mediators, but how their interaction contributes to persistent pain is unknown. Here, we identify Tmem100 as a potentiating modulator of TRPA1-V1 complexes. Tmem100 is coexpressed and forms a complex with TRPA1 and TRPV1 in DRG neurons. Tmem100-deficient mice show a reduction in inflammatory mechanical hyperalgesia and TRPA1- but not TRPV1-mediated pain. Single-channel recording in a heterologous system reveals that Tmem100 selectively potentiates TRPA1 activity in a TRPV1-dependent manner. Mechanistically, Tmem100 weakens the association of TRPA1 and TRPV1, thereby releasing the inhibition of TRPA1 by TRPV1. A Tmem100 mutant, Tmem100-3Q, exerts the opposite effect; i.e., it enhances the association of TRPA1 and TRPV1 and strongly inhibits TRPA1. Strikingly, a cell-permeable peptide (CPP) containing the C-terminal sequence of Tmem100-3Q mimics its effect and inhibits persistent pain. Our study unveils a context-dependent modulation of the TRPA1-V1 complex, and Tmem100-3Q CPP is a promising pain therapy.


Asymmetric inheritance of centrosomes maintains stem cell properties in human neural progenitor cells.
During human forebrain development, neural progenitor cells (NPCs) in the ventricular zone (VZ) undergo asymmetric cell divisions to produce a self-renewed progenitor cell, maintaining the potential to go through additional rounds of cell divisions, and differentiating daughter cells, populating the developing cortex. Previous work in the embryonic rodent brain suggested that the preferential inheritance of the pre-existing (older) centrosome to the self-renewed progenitor cell is required to maintain stem cell properties, ensuring proper neurogenesis. If asymmetric segregation of centrosomes occurs in NPCs of the developing human brain, which depends on unique molecular regulators and species-specific cellular composition, remains unknown. Using a novel, recombination-induced tag exchange-based genetic tool to birthdate and track the segregation of centrosomes over multiple cell divisions in human embryonic stem cell-derived regionalised forebrain organoids, we show the preferential inheritance of the older mother centrosome towards self-renewed NPCs. Aberration of asymmetric segregation of centrosomes by genetic manipulation of the centrosomal, microtubule-associated protein Ninein alters fate decisions of NPCs and their maintenance in the VZ of human cortical organoids. Thus, the data described here use a novel genetic approach to birthdate centrosomes in human cells and identify asymmetric inheritance of centrosomes as a mechanism to maintain self-renewal properties and to ensure proper neurogenesis in human NPCs.


Estimating time-varying nonlinear autoregressive model parameters by minimizing hypersurface distance.
A nonleast-squares (non-LS) based method is presented for modeling time-varying (TV) nonlinear systems. The proposed method combines basis function technique and minimization of hypersurface distance (MHD) to combat TV and nonlinear dynamics, respectively. The performance of TVMHD is compared to the LS and total LS methods using simulation examples as well as human heart rate data recorded during different body positions. With all data, TVMHD significantly outperforms the two other methods by a factor of one order of magnitude; the LS-based methods require double the number of parameters than TVMHD requires to obtain similar residual error values. The significance of TVMHD is that due to its accurate parameter estimates concomitant with a fewer number of parameters, we now have the possibility of pinpointing parameters that may be of physiological importance, where such application will be especially useful in discriminating diseased conditions. Furthermore, our algorithm allows discrimination of model terms, which are TV or time invariant, by examining those basis function coefficients that are designed to capture TV dynamics. However, it should be noted that the main disadvantage of TVMHD is that it requires significantly greater computational time than the LS-based methods.


COVID-19 outbreak control, example of ministry of health of Turkey
Our first COVID-19 case in Turkey was a 44-year-old male who referred to the hospital on March 9, 2020. The first related death occurred on March 17, 2020. Preparedness for the pandemic has been ongoing before the first case was detected. The National Pandemic Plan was published in 2006. The Pandemic Influenza National Preparedness Plan was available after being updated in light of experiences gained during the 2009 Influenza A pandemic. Accordingly, Pandemic Coordination Boards and Operation Centers have been established on the national and provincial levels. This was an adaptable plan to the Novel Coronavirus Disease (COVID-19). We formed teams to work on a 24/7 basis and established a Scientific Committee at the Public Health Emergency Operation Center within the General Directorate of Public Health. "COVID-19 Risk Assessment", "COVID-19 Guideline" and "Case Report Form", regulations of personal protective equipment along with need-based guidelines, treatment algorithms, brochures and related documents have been released. For the case-based follow-up, Public Health Management System (HSYS) is being used. PCR and rapid diagnostic kits are being used to analyze the samples at the central Microbiology Reference Laboratory and the authorized laboratories in several provinces. Various preventive measures were implemented including flight restrictions to certain countries, gradually expanded to suspending all flights and prohibiting the entry of foreign nationals, 14-day isolation and symptom monitoring for those that came from countries under risk. Persons with chronic diseases have been granted an administrative leave, on campus education at schools and activities of public rest and entertainment areas were temporarily suspended. The measures have been implemented for penitentiary institutions, dormitories, nursing homes, public transport and intercity buses, and also weekend curfews are implemented. In accordance with the pandemic plan, actions have been carried out with a multi-sectoral approach, and preventive measures have been implemented to cover the society as a whole.


[Solearhynchus rhytidotes nov. comb. (Palaeacanthocephala: Echinorhynchidae), a parasite of soles (Soleidae)].
On the base of literature and the original data in has been established that acanthocephalan species known as Acanthocephaloides kostylewi Meyer, 1933, and described from the Black Sea sole Solea lascaris nasuta (Pallas) is to be transferred to the genus Solearhynchus with new species name Solearhynchus rhytidotes nov. comb. (syn. Echinorhynchus rhytidotes Monticelli, 1905; E. aurantiacus Monticelli, 1887; E. corrugatus Monticelli, 1887; Acanthocephaloides kostylewi Meyer, 1933; A. rhytidotes (Monticelli, 1904) Belofastova et Korniychuc, 2000; Solearhynchus soleae (Porta, 1905) de Buron et Moillard, 1985). Detailed descriptions, measurements and original figures are given.


Risk of Cardiovascular Complication Among Type 2 Diabetes Mellitus Patients in Medan, Indonesia. A Cross-sectional Study.
Cardiovascular disease (CVD) is a disease associated with impaired heart and blood vessel function. The occurrence of Diabetes Mellitus (DM) is due to an imbalance in blood glucose that could cause damage to large and small blood vessels and increase platelet aggregation, contributing to atherosclerosis and cardiovascular disease. It could be attributed to high blood pressure, smoking, diabetes, lack of movement, obesity, high cholesterol, unbalanced diet. CVD can be prevented, especially in at-risk groups, with risk assessment utilizing the Framingham Risk Score (FRS). The study aimed to to analyze the risk of CVD according to the Framingham Risk Score (FRS) in type 2 DM patients in Medan. The data were collected with the Framingham Risk Score, which consisted of sub-variables of age, total cholesterol, HDL cholesterol, systolic blood pressure (SBM), history of DM, and history of smoking, which were assessed by gender. It was a cross-sectional analytic study with consecutive sampling on 252 respondents, namely DM patients who came for treatment or control to the health centers in Medan and fulfilled the inclusion and exclusion criteria. The data were then analyzed with SPSS. The respondents consisted of 197 women (78.2%) and 55 men (21.8%). The results were predominated with high risk patients (139, 55.2%), followed by those with medium risks (80, 31.7%), and low risks (33, 13.1%). Chi-square test yielded a significant association between each sub-variable of CVD risk factors and the risks based on FRS in type 2 DM patients. Each sub-variable of CVD risk factors and the risks on FRS among type 2 DM patients. In future studies, it is recommended to include that a larger number of samples in the community with a balanced proportion of men and women.


Sports betting incentives encourage gamblers to select the long odds: An experimental investigation using monetary rewards.
Incentives for wagering products can provide extra value to gamblers. However, there is no financial reason why this added value should lead people to take greater gambling risks. This study aimed to experimentally test if wagering incentives cause gamblers to choose higher-risk (long odds) bets than un-incentivized bets. An online experiment was conducted with wagering customers (N = 299, female = 12). Participants bet $4 on each of six video game simulations of a sport that they had wagered on in the past 12 months (Australian Football League, Cricket, or Soccer). Each game offered different common wagering incentives: Bonus bet, Better odds/winnings, Reduced risk, Cash rebate, Player's choice of inducement, or No-inducement. For each game, participants could bet on long, medium, or short odds, and subsequently viewed a highlight reel of the simulated game outcome and bet outcome. Participants selected significantly longer odds (i.e., riskier) bets on games when an incentive was offered compared to the No-inducement condition. Better odds/winnings was the most attractive incentive, followed by Bonus bet, Cash rebate, Reduced risk, and No-incentive, respectively. No significant differences were observed based on demographics or problem gambling severity. The choice of long odds with incentivized bets increases the volatility of player returns. Increased volatility results in more gamblers in a losing position and fewer gamblers with larger wins. Moreover, if long odds bets are priced to provide poorer value to bettors compared to short odds, they would increase gamblers' losses and equivalently increase operators' profits.


The ABCs for deciding on a decision support system in the health care industry.
As the information superhighway becomes clustered with various organizations sharing the hope of staying solvent with the aid of technology, managers in the health care industry are more and more looking to efficient Decision Support Systems (DSS). The focus of DSS selection has often been on evaluations that measure decision performance for determining the influence and efficacy of the tool. Such evaluations, in many instances, ignore pertinent measurements for a successful selection and implementation of a DSS. In this article, the authors present a simple but important set of evaluation factors that can "make or break" a DSS implementation in the health care industry.


[Severe post-COVID-19 dialysis dependence and inpatient acute kidney injury].
COVID-19 challenged our health system, within the broad clinical spectrum acute kidney injury was presented as a catastrophic event, acute kidney injury and the risk of dependency after dialysis constitute a clinical problem with high repercussions in the funcionality. To identify risk factors for dialysis dependence after acute kidney injury from COVID-19. A retrospective observational cohort study was carried out at the Hospital de Especialidades del Centro Médico Nacional Siglo XXI, of the Mexican Institute of Social Security, from March 2020 to March 2021. 317 patients were included, we performed descriptive statistics, we compared differences between the stages of acute kidney injury, finding a difference in obesity with a frequency of 2.2% in stage 1, 20.82% stage 2 and 14.51% stage 3, with p value = 0.018. We found dialysis dependence one year after hospital-acquired acute kidney injury induced by COVID-19 in 58 patients (18.9%), we analyzed by KDIGO stage, in those patients who had AKI KDIGO 1 (2.83%) it depended on dialysis at one year, in the KDIGO stage 2 (3.78%), in the KDIGO stage 3 (11.67%). Our study allowed us to identify that the risk factors associated with dialysis dependence are: male gender, type 2 diabetes mellitus, obesity, cardiovascular disease.


Phase I dose escalation study and pilot efficacy analysis of LXI-15029, a novel mTOR dual inhibitor, in Chinese subjects with advanced malignant solid tumors.
The mammalian target of rapamycin (mTOR) kinase, a central component of the PI3K/AKT/mTOR pathway, plays a critical role in tumor biology as an attractive therapeutic target. We conducted this first-in-human study to investigate the safety, pharmacokinetics (PK), and pilot efficacy of LXI-15029, an mTORC1/2 dual inhibitor, in Chinese patients with advanced malignant solid tumors. Eligible patients with advanced, unresectable malignant solid tumors after failure of routine therapy or with no standard treatment were enrolled to receive ascending doses (10, 20, 40, 60, 80, 110, and 150 mg) of oral LXI-15029 twice daily (BID) (3 + 3 dose-escalation pattern) until disease progression or intolerable adverse events (AEs). The primary endpoints were safety and tolerability. Between June 2017 and July 2021, a total of 24 patients were enrolled. LXI-15029 was well tolerated at all doses. Only one dose-limiting toxicity (grade 3 increased alanine aminotransferase) occurred in the 150 mg group, and the maximum tolerated dose was 110 mg BID. The most common treatment-related AEs were leukocytopenia (41.7%), increased alanine aminotransferase (20.8%), increased aspartate aminotransferase (20.8%), prolonged electrocardiogram QT interval (20.8%), and hypertriglyceridemia (20.8%). No other serious treatment-related AEs were reported. LXI-15029 was absorbed rapidly after oral administration. The increases in the peak concentration and the area under the curve were greater than dose proportionality over the dose range. Eight patients had stable disease. The disease control rate was 40.0% (8/20; 95% CI 21.7-60.6). In evaluable patients, the median progression-free survival was 29 days (range 29-141). LXI-15029 demonstrated reasonable safety and tolerability profiles and encouraging preliminary antitumor activity in Chinese patients with advanced malignant solid tumors, which warranted further validation in phase II trials. NCT03125746(24/04/2017), http://ClinicalTrials.gov/show/NCT03125746.


Distance measures and evolution of polymer chains in their topological space.
Conformational transitions are ubiquitous in biomolecular systems, have significant functional roles and are subject to evolutionary pressures. Here we provide a first theoretical framework for topological transition, i.e. conformational transitions that are associated with changes in molecular topology. For folded linear biomolecules, arrangement of intramolecular contacts is identified as a key topological property, termed as circuit topology. Distance measures are proposed as reaction coordinates to represent progress along a pathway from initial topology to final topology. Certain topological classes are shown to be more accessible from a random topology. We study dynamic stability and pathway degeneracy associated with a topological reaction and found that off-pathways might seriously hamper evolution to desired topologies. Finally we present an algorithm for estimating the number of intermediate topologies visited during a topological reaction. The results of this study are relevant to, among others, structural studies of RNA and proteins, analysis of topologically associated domains in chromosomes, and molecular evolution.


Immune response in preleukemic mice.
Humoral and cellular immunity was assessed serially in preleukemic AKR mice and Gross virus-injected C3H/HeJ mice over a period of 10 to 12 months. Quantitative gamma globulin, hemagglutinin and hemolysin titers after immunization with sheep red blood cells, and macrophage-migration inhibition were determined. Normal production of gamma globulins as well as humoral antibodies was found in both of these experimental groups of mice, whereas macrophage-migration inhibition, which is believed to be a correlate of cellular immunity, appeared to be abnormal throughout the life of these animals, which were destined to develop a high incidence of lymphoma. Some aspects of this apparent dissociation between humoral and cellular immune response are discussed with regard to the mouse virus lymphoma system, primarily involving the thymus, which was used in these studies.


Aqua-{2-(pyridin-2-yl)-N-[(pyridin-2-yl)methyl-idene]ethanamine-κ(3) N,N',N''}(sulfato-κ(2) O,O')copper(II) tetra-hydrate.
The title complex, [Cu(SO4)(C13H13N3)(H2O)]·4H2O, was obtained by mixing copper sulfate penta-hydrate and 2-(pyridin-2-yl)-N-(pyridin-2-yl-methyl-idene)ethanamine in eth-anol under reflux conditions. The Cu(II) ion shows a Jahn-Teller-distorted octa-hedral geometry, with equatorial positions occupied by three N atoms from the tridentate ligand (average Cu-N = 2.004 Å) and one O atom from a bidentate sulfate anion [Cu-O = 1.963 (2) Å]. The axial positions are occupied by one O atom from a coordinating water mol-ecule [Cu-O = 2.230 (3) Å] and one weakly bonded O atom [Cu-O = 2.750 (2) Å] from the bidentate sulfate ion. The complex mol-ecules are connected through O-H⋯O hydrogen bonds between the coordinating water mol-ecules and sulfate ions from neighboring complexes, forming a double chain parallel to the c axis. The chains are stabilized through additional hydrogen bonds by one of the non-coordinating water mol-ecules bridging between neighboring strands of the double chains. The remaining three water mol-ecules fill the inter-stitial space between the double chains and are involved in an intricate hydrogen-bonding network that consolidates the structure.


A double-blind pilot study of the effect of Prokarin on fatigue in multiple sclerosis.
In this 12-week study with 29 subjects, the effect of Prokarin (n=22), a proprietary blend of histamine and caffeine, was compared to placebo group (n=7) for the following outcomes: 1) fatigue as measured by the Modified Fatigue Impact Scale (MFIS); 2) lower limb function as measured by timed walk test; 3) upper limb function as measured by the pegboard test; 4) cognitive function as measured by the Paced Auditory Serial Additions Test (PASAT); 5) serum caffeine level; 6) change in brain chemistry as measured by quantitative magnetic resonance spectroscopy assay of N-acetyl aspartate (NAA); and 7) safety as measured by routine blood chemistry, TSH and urinalysis. Data were acquired at baseline, 4, 8 and 12 weeks. The Prokarin group MFIS mean was significantly different rom the mean of the placebo group at 12 weeks (df=24, t=2.08, P=<0.02), with respective means of 37.40, SD=15.18, for the Prokarin group and 53.2, SD=11.39 for the controls. For the secondary endpoints (PASAT, 25 foot timed walk, peg test, and magnetic resonance spectroscopy [MRS]), there were no significant differences between the Prokarin-treated group and the placebo group. However, there were significant improvements within the Prokarin group for each of these measures for the pre- versus posttreatment comparison at 12 weeks. Serum caffeine data indicated that caffeine exerted no independent effect on performance. No laboratory abnormalities were seen, and the treatment was well tolerated. There was a modest-size statistical effect of Prokarin on fatigue in multiple sderosis (MS) compared with the placebo group. A larger trial is warranted, based on this pilot study.


[Toll-like receptor 4 expression and function of respiratory syncytial virus-infected airway epithelial cells].
To observe the epithelial Toll like receptor (TLR)4 expression changes and the signaling pathway function after respiratory syncytial virus (RSV) infection and to explore the mechanisms of RSV-induced airway inflammation. 9HTEo-human tracheal epithelial cell line was infected by RSV (MOI = 10), and TLR1-10 mRNA were detected by RT-PCR assay at 3 h post RSV infection. TLR4 mRNA was detected by real time Q-PCR assay at 3 h, 6 h and 9 h post RSV infection, and TLR4 protein expression and cell apoptosis were determined by flow cytometry at 24 h post RSV infection. IL-8 in supernatant was detected by ELISA after RSV-infected cells exposed to lipopolysaccharide (LPS). A normal control group and a RSV infection group were set up for the RT-PCR and flow cytometry experiments, and the data were analyzed by paired t test using GraphPad 4.0 software. A normal group, a RSV group and a UV-inactivated RSV group were set up for the real time Q-PCR, experiments, and the data were analyzed by Kruskal-Wallis test. The ELISA experiments were divided into 4 groups including a normal control, a RSV, a LPS stimulation, and a RSV plus LPS co-stimulation groups, and the data were analyzed by One-way ANOVA test. (1) TLR2-10 mRNA level was significantly up-regulated (t value of TLR2-10: 3.49 -14.47, P < 0.05), especially TLR-2, 6 enhanced expression, compared with the normal epithelial cells. Real time Q-PCR assay showed that TLR4 mRNA started to increase at 3hr (Kruskal-Wallis test value = 8.82, P < 0.05, n = 6) and significantly elevated at 9 hour (Kruskal-Wallis test value = 6.62, P < 0.05, n = 6). UV inactivated-RSV had no effect on the TLR4 mRNA level. (2) Flow cytometry showed that membrane TLR4 mean fluorescence intensity (MFI) increased (RSV: 1.27 +/- 0.48, normal: 0.97 +/- 0.25; t = 2.39, P > 0.05, n = 10) while cytoplasmic TLR4 MFI simultaneously decreased (RSV: 3.08 +/- 1.38, normal: 3.36 +/- 1.31, t = 2.92, P = 0.225, n = 10). Percentage of membrane TLR4-positive cells was higher in RSV infected population [RSV: (11.99 +/- 7.74)%, normal: (1.16 +/- 0.47)%, Mann-Whitney t value = 0.001, P < 0.01, n = 8], most (93.32 +/- 1.7)% of which were Annexin V positive. IL-8 was significantly induced in the RSV plus LPS costimulation group compared with RSV group (F = 59.29, P < 0.01, n = 3). RSV induced epithelial TLR4 up-regulation, localization changes from cytoplasm to membrane, IL-8 secretion through TLR4 signaling pathway and epithelial cell apoptosis in membrane TLR4 positive population. These results indicate TLR4 is involved in RSV-induced acute or chronic epithelial-dependent inflammation, which might contribute to acute or chronic airway inflammation.


Unlocking loxP to Track Genome Editing In Vivo.
The development of CRISPR-associated proteins, such as Cas9, has led to increased accessibility and ease of use in genome editing. However, additional tools are needed to quantify and identify successful genome editing events in living animals. We developed a method to rapidly quantify and monitor gene editing activity non-invasively in living animals that also facilitates confocal microscopy and nucleotide level analyses. Here we report a new CRISPR "fingerprinting" approach to activating luciferase and fluorescent proteins in mice as a function of gene editing. This system is based on experience with our prior cre recombinase (cre)-detector system and is designed for Cas editors able to target loxP including gRNAs for SaCas9 and ErCas12a. These CRISPRs cut specifically within loxP, an approach that is a departure from previous gene editing in vivo activity detection techniques that targeted adjacent stop sequences. In this sensor paradigm, CRISPR activity was monitored non-invasively in living cre reporter mice (FVB.129S6(B6)-Gt(ROSA)26Sortm1(Luc)Kael/J and Gt(ROSA)26Sortm4(ACTB-tdTomato,-EGFP)Luo/J, which will be referred to as LSL-luciferase and mT/mG throughout the paper) after intramuscular or intravenous hydrodynamic plasmid injections, demonstrating utility in two diverse organ systems. The same genome-editing event was examined at the cellular level in specific tissues by confocal microscopy to determine the identity and frequency of successfully genome-edited cells. Further, SaCas9 induced targeted editing at efficiencies that were comparable to cre, demonstrating high effective delivery and activity in a whole animal. This work establishes genome editing tools and models to track CRISPR editing in vivo non-invasively and to fingerprint the identity of targeted cells. This approach also enables similar utility for any of the thousands of previously generated loxP animal models.


A rapid liquid biopsy of lung cancer by separation and detection of exfoliated tumor cells from bronchoalveolar lavage fluid with a dual-layer "PERFECT" filter system.
Separation and detection of exfoliated tumor cells (ETCs) from bronchoalveolar lavage fluid (BALF), namely the liquid biopsy of BALF, has been proved to be a valuable tool for the diagnosis of lung cancer. Herein, we established a rapid liquid biopsy of BALF based on a dual-layer PERFECT (precise, efficient, rapid, flexible, easy-to-operate, controllable and thin) filter system for the first time. Methods: The dual-layer PERFECT filter system consists of an upper-layer filter with large micropores (feature size of 49.4 ± 0.5 μm) and a lower-layer filter with small micropores (9.1 ± 0.1 μm). The upper-layer filter contributes to the isolation of cell clusters and removal of mucus from BALF samples, meanwhile the lower-layer one targets for the separation of single ETCs. First, separation of 10000 spiked A549s (cultured lung cancer cells) from 10 mL clinical BALF samples (n=3) were performed to investigate the performance of the proposed system in rare cell separation. Furthermore, separation and detection of ETCs and ETC clusters from clinical BALF samples were performed with this system to test its efficacy and compared with the routine cytocentrifuge. The clinical BALF samples were collected from 33 lung cancer-suspected patients with visible lesions under bronchoscope. The final histopathological results showed that 20 samples were from lung cancer positive patients while the other 13 cases were from lung cancer negative patients. Results: The recovery rate of spiked A549 cells from clinical BALF samples with the developed system (89.8 ± 5.2%) is significantly higher than that with the cytocentrifuge (13.6 ± 7.8%). In the preliminary clinical trial, although 33 clinical BALF samples with volume ranging from 6 mL to 18 mL showed greatly varied turbidity, filtrations could be finished within 3 min for 54.6% of samples (18/33), and 10 min at most for the rest. The dual-layer PERFECT filter system is proved to have a much higher sensitivity (80.0%, 95% CI: 55.7%-93.4%) than the routine cytocentrifuge (45.0%, 95% CI: 23.8%-68.0%), p=0.016 (McNemar test, two-tail). Moreover, the sensitivity of this platform is neither interfered by the variations of turbidity of the BALF samples, nor associated with the types of lung cancer. Conclusions: The easy and rapid processing of BALF samples with varying volume and turbidity, competitive sensitivity and good versatility for different lung cancer types will make the established dual-layer PERFECT filter system a promising approach for the liquid biopsy of BALF. The high-performance BALF-based liquid biopsy will improve the cytopathological identification and diagnosis of lung cancer.


Multilevel predictors of clinic adoption of state-supported trainings in children's services.
Characteristics associated with participation in training in evidence-informed business and clinical practices by 346 outpatient mental health clinics licensed to treat youths in New York State were examined. Clinic characteristics extracted from state administrative data were used as proxies for variables that have been linked with adoption of innovation (extraorganizational factors, agency factors, clinic provider-level profiles, and clinic client-level profiles). Multiple logistic regression models were used to assess the independent effects of theoretical variables on the clinics' participation in state-supported business and clinical trainings between September 2011 and August 2013 and on the intensity of participation (low or high). Interaction effects between clinic characteristics and outcomes were explored. Clinic characteristics were predictive of any participation in trainings but were less useful in predicting intensity of participation. Clinics affiliated with larger (adjusted odds ratio [AOR]=.65, p<.01), more efficient agencies (AOR=.62, p<.05) and clinics that outsourced more clinical services (AOR=.60, p<.001) had lower odds of participating in any business-practice trainings. Participation in business trainings was associated with interaction effects between agency affiliation (hospital or community) and clinical staff capacity. Clinics with more full-time-equivalent clinical staff (AOR=1.52, p<.01) and a higher proportion of clients under age 18 (AOR=1.90, p<.001) had higher odds of participating in any clinical trainings. Participating clinics with larger proportions of youth clients had greater odds of being high adopters of clinical trainings (odds ratio=1.54, p<.01). Clinic characteristics associated with uptake of business and clinical training could be used to target state technical assistance efforts.


Stage III and Metastatic Lymph Node Ratio Are the only Independent Prognostic Factors in Colorectal Signet-ring Cell Carcinoma Patients.
Signet-ring cell carcinoma (SRCC) is an uncommon histological variant of colorectal cancer (CRC). Knowledge is scarce due to its rarity. Our aim was to better evaluate the clinicopathologic and prognostic features of this little-known malignancy. Thirty-nine consecutive patients with non-metastatic colorectal SRCC undergoing curative resection at University Hospital of Parma between 2000 and 2018 were examined in this retrospective analysis. Mean overall (OS) and disease-free survival (DFS) were 33.6 and 31.5 months, respectively. At univariate analysis, the lymph-related parameters (nodal status, Stage III, metastatic lymph node ratio and lymphovascular invasion) were significantly associated with shorter OS and poorer DFS. At multivariate analysis, Stage III and a metastatic lymph node ratio ≥25% were found to be the only independent prognostic factors significantly correlated with worse OS and DFS. Nodal and lymphatic status should be carefully pondered when planning the most appropriate management of patients with colorectal SRCC.


Picornaviral polymerase domain exchanges reveal a modular basis for distinct biochemical activities of viral RNA-dependent RNA polymerases.
Picornaviral RNA-dependent RNA polymerases (RdRPs) have low replication fidelity that is essential for viral fitness and evolution. Their global fold consists of the classical "cupped right hand" structure with palm, fingers, and thumb domains, and these RdRPs also possess a unique contact between the fingers and thumb domains. This interaction restricts movements of the fingers, and RdRPs use a subtle conformational change within the palm domain to close their active sites for catalysis. We have previously shown that this core RdRP structure and mechanism provide a platform for polymerases to fine-tune replication rates and fidelity to optimize virus fitness. Here, we further elucidated the structural basis for differences in replication rates and fidelity among different viruses by generating chimeric RdRPs from poliovirus and coxsackievirus B3. We designed these chimeric polymerases by exchanging the fingers, pinky finger, or thumb domains. The results of biochemical, rapid-quench, and stopped-flow assays revealed that differences in biochemical activity map to individual modular domains of this polymerase. We found that the pinky finger subdomain is a major regulator of initiation and that the palm domain is the major determinant of catalytic rate and nucleotide discrimination. We further noted that thumb domain interactions with product RNA regulate translocation and that the palm and thumb domains coordinately control elongation complex stability. Several RdRP chimeras supported the growth of infectious poliovirus, providing insights into enterovirus species-specific protein-protein interactions required for virus replication.


[Clinical underestimation of the prevalence of approximal dentin lesions and inadequate restorations].
The aim of this study was to determine the additional diagnostic value of bitewing radiographs compared to clinical examination, with respect to the presence of dentine lesions and inadequate restorations for different age groups. Subjects were 14-, 17-, 20-, 23-years-old, or were in the age groups of 25-34 and 35-54 years, who participated in 2 clinical epidemiological surveys. After obtaining informed consent, bitewing radiographs were made of approximately 25% of the participants (n = 663). The extra diagnostic yield of the bitewing radiographs varied between a factor 1.6 and 7.2 for approximal dentine caries diagnosis of untreated surfaces, and between 2.3 and 5.9 for inadequate restorations. In conclusion, a considerable increase in the numbers of approximal dentinal lesions and inadequate restorations for all age groups was found.


R Loops: From Physiological to Pathological Roles.
DNA-RNA hybrids play a physiological role in cellular processes, but often, they represent non-scheduled co-transcriptional structures with a negative impact on transcription, replication and DNA repair. Accumulating evidence suggests that they constitute a source of replication stress, DNA breaks and genome instability. Reciprocally, DNA breaks facilitate DNA-RNA hybrid formation by releasing the double helix torsional conformation. Cells avoid DNA-RNA accumulation by either preventing or removing hybrids directly or by DNA repair-coupled mechanisms. Given the R-loop impact on chromatin and genome organization and its potential relation with genetic diseases, we review R-loop homeostasis as well as their physiological and pathological roles.


Magnetic Resonance Elastography of Rodent Brain.
Magnetic resonance elastography (MRE) is a non-invasive imaging technique, using the propagation of mechanical waves as a probe to palpate biological tissues. It consists in three main steps: production of shear waves within the tissue; encoding subsequent tissue displacement in magnetic resonance images; and extraction of mechanical parameters based on dedicated reconstruction methods. These three steps require an acoustic-frequency mechanical actuator, magnetic resonance imaging acquisition, and a post-processing tool for which no turnkey technology is available. The aim of the present review is to outline the state of the art of reported set-ups to investigate rodent brain mechanical properties. The impact of experimental conditions in dimensioning the set-up (wavelength and amplitude of the propagated wave, spatial resolution, and signal-to-noise ratio of the acquisition) on the accuracy and precision of the extracted parameters is discussed, as well as the influence of different imaging sequences, scanners, electromagnetic coils, and reconstruction algorithms. Finally, the performance of MRE in demonstrating viscoelastic differences between structures constituting the physiological rodent brain, and the changes in brain parameters under pathological conditions, are summarized. The recently established link between biomechanical properties of the brain as obtained on MRE and structural factors assessed by histology is also studied. This review intends to give an accessible outline on how to conduct an elastography experiment, and on the potential of the technique in providing valuable information for neuroscientists.


Comparison of hyperkalemic risk in hospitalized patients treated with different angiotensin receptor blockers: a retrospective cohort study using a Korean clinical research database.
Clinical use of angiotensin II type 1 receptor antagonists (angiotensin receptor blockers [ARBs]) is associated with hyperkalemia as an adverse drug reaction. Although it has significant clinical implications, the incidence and relative risks of hyperkalemia with various ARBs have not yet been fully evaluated. The purpose of this study was to determine the risk of hyperkalemic events in hospitalized patients treated with different ARBs and to compare the risk among them. We constructed a retrospective cohort composed of hospitalized adult patients who took ARBs in a single tertiary teaching hospital between April 2004 and March 2010. We estimated the incidence of hyperkalemia (serum potassium level >5.5 mEq/L) with various ARBs, and then compared the risk between them using a multivariate Cox proportional hazard model based on age, sex, Charlson co-morbidity score, baseline serum potassium, underlying diseases, and concomitant drugs. We identified 6992 evaluable intervals from 5449 patients treated with one of the seven ARBs during hospitalization over the 71-month study period with 2521.6 patient-months. We found 381 hyperkalemic events (5.4%) during the study period and an overall event rate of 15.1/100 patient-months. Moderate to fatal hyperkalemia was relatively rare (>6.0 mEq/L, 2.1% [moderate]; >6.5 mEq/L, 0.9% [severe]; >7.0 mEq/L, 0.3% [fatal]). After adjustment for covariates, telmisartan showed a lower risk of hyperkalemia (hazard ratio 0.67; 95% confidence interval 0.51, 0.89) compared with all other ARBs. The risk of hyperkalemic events in hospitalized patients treated with different ARBs was defined. Telmisartan showed a relatively lower hyperkalemic risk profile in hospitalized patients compared with other ARBs.


Biodiversity of Asian rice gall midge (Orseolia oryzae Wood Mason) from five countries examined by AFLP analysis.
Amplified fragment length polymorphism (AFLP) analysis was used to assess the biodiversity of one of the most important dipteran pests of cereals, the Asian rice gall midge (Orseolia oryzae Wood Mason). Larvae and pupae were collected at 15 locations in five Asian countries and preserved in 95% ethanol for storage, shipment, and DNA extraction using cetyltrimethylammonium bromide (CTAB). Although only approximately 1 microg of DNA was extracted from a single pupa or larva, the use of several AFLP primers in various combinations meant that this amount of DNA was sufficient to allow many DNA fingerprints to be made per individual. Fingerprints were sufficiently reproducible, especially during selective amplification, to allow the genetic diversity within a field population to be characterized. Extraction of DNA from a pool of 20 insects yielded AFLP fingerprints in which variation among individuals was sacrificed in favor of detecting differences among populations. For each location, pooled DNA was amplified with three primer pairs. A total of 261 distinct AFLP bands were identified for the 45 fingerprints. Cluster analysis, performed by the unweighted pair-group method (UPGMA), separated the populations into two distinct groups. Group I included two populations from Guangdong province of southern China and one each from Laos and Imphal in northeastern India, while group II was comprised of eleven populations from elsewhere in India (Assam, Orissa, Madhya Pradesh, Andhra Pradesh, and Kerala) and from Nepal and Sri Lanka. AFLP analysis provided insight into the origins of gall midge biotypes. In 1992, the prevailing biotype in Imphal changed from Indian biotype 3 to a new biotype 3M. Our data show that biotype 3M belongs to group I and did not arise by a recent mutation from biotype 3, which belongs to group II. By contrast, Indian biotypes 2 and 4 are likely to have diverged through recent mutation and selection, as are Chinese biotypes 1 and 4. The almost simultaneous emergence of new biotypes in Kerala and Sri Lanka during 1985-1988 was most probably coincidental, because these biotypes are not closely related. AFLP fingerprints were also able to detect sexual dimorphism in the DNA of adult gall midges and to distinguish gall midge from its major parasite Platygaster oryzae.


A small-RNA-mediated negative feedback loop controls quorum-sensing dynamics in Vibrio harveyi.
The bioluminescent marine bacterium Vibrio harveyi uses a cell-to-cell communication process called quorum sensing (QS) to co-ordinate behaviours in response to changes in population density. QS is accomplished through the secretion and detection of extracellular signalling molecules called autoinducers. At the centre of the V. harveyi QS circuit are five small regulatory RNAs called Qrr1-5 which destabilize the mRNA of luxR, encoding LuxR, the master transcriptional regulator of QS target genes. Here we show that LuxR directly activates transcription of qrr2, qrr3 and qrr4, leading to the rapid downregulation of luxR. The LuxR-binding sites in the promoters of qrr2, qrr3 and qrr4 were identified and mutated to determine the consequences of this regulatory loop on QS dynamics. Disruption of the loop delays the transition from high to low cell density, and more significantly, decreases the cell density at which the population reaches a quorum. Our results suggest that feedback is essential for optimizing the dynamics of the transitions between individual and group behaviours.


Health of herring gulls (Larus argentatus) in relation to breeding location in the early 1990s. III. Effects on the bone tissue.
Health effects associated with the Great Lakes environment were assessed in adult herring gulls (Larus argentatus) in the early 1990s, including the size and quality of their bones. Femurs were excised from 140 individuals from 10 colonies distributed throughout the Great Lakes and 2 reference colonies in Lake Winnipeg (freshwater) and the Bay of Fundy (marine). Femurs of gulls from the Great Lakes differed from the freshwater or marine reference for 9 of 12 variables of size, composition, and strength assessed using peripheral quantitative computed tomography (pQCT) and biomechanical testing. Femurs of Great Lakes gulls were significantly smaller in length (-2.9%), periosteal circumference (-2.4%), and cross-sectional area (-5.4%) than freshwater reference birds. Femurs of the Great Lakes gulls had a lower significant cortical bone mineral content (-8.1%) and density (-2%) than the marine reference. A significant increase in the amount the bone could bend before it broke (+34%) and the energy required to break it (+44%) and a significant decrease (-16.3%) in stiffness during three-point biomechanical bending test were also detected in Great Lakes versus the freshwater gulls. These differences are indicative of impaired mineralization. When divided into high and low 2,3,7,8-tetrachlorodibenzo-p-dioxin toxicity equivalent (TCDD-TEQ) colonies, the amount the bone could bend before it broke and the energy required to break it were significantly higher in the high TEQ colonies, but not high polychlorinated biphenyl (PCB) colonies. Breeding location and dietary choices of Great Lakes herring gulls in the early 1990s resulted in modulations of physiological processes that affected the size, mineralization, and biomechanical properties of bone.


Changes in the levels of inositol phosphates after agonist-dependent hydrolysis of membrane phosphoinositides.
The formation of inositol phosphates in response to agonists was studied in brain slices, parotid gland fragments and in the insect salivary gland. The tissues were first incubated with [3H]inositol, which was incorporated into the phosphoinositides. All the tissues were found to contain glycerophosphoinositol, inositol 1-phosphate, inositol 1,4-bisphosphate and inositol 1,4,5-trisphosphate, which were identified by using anion-exchange and high-resolution anion-exchange chromatography, high-voltage paper ionophoresis and paper chromatography. There was no evidence for the existence of inositol 1:2-cyclic phosphate. A simple anion-exchange chromatographic method was developed for separating these inositol phosphates for quantitative analysis. Stimulation caused no change in the levels of glycerophosphoinositol in any of the tissues. The most prominent change concerned inositol 1,4-bisphosphate, which increased enormously in the insect salivary gland and parotid gland after stimulation with 5-hydroxytryptamine and carbachol respectively. Carbachol also induced a large increase in the level of inositol 1,4,5-trisphosphate in the parotid. Stimulation of brain slices with carbachol induced modest increase in the bis- and tris-phosphate. In all the tissues studied, there was a significant agonist-dependent increase in the level of inositol 1-phosphate. The latter may be derived from inositol 1,4-bisphosphate, because homogenates of the insect salivary gland contain a bisphosphatase in addition to a trisphosphatase. These results suggest that the earliest event in the stimulus-response pathway is the hydrolysis of polyphosphoinositides by a phosphodiesterase to yield inositol 1,4,5-trisphosphate and inositol 1,4-bisphosphate, which are subsequently hydrolysed to inositol 1-phosphate and inositol. The absence of inositol 1:2-cyclic phosphate could indicate that, at very short times after stimulation, phosphatidylinositol is not catabolized by its specific phosphodiesterase, or that any cyclic derivative liberated is rapidly hydrolysed by inositol 1:2-cyclic phosphate 2-phosphohydrolase.


Bullying and sexual harassment of junior doctors in New South Wales, Australia: rate and reporting outcomes.
Objective The aim of this study was to describe rates of exposure to bullying and sexual harassment in junior doctors in first- or second-year prevocational medical training (PGY1 or PGY2 respectively) positions in New South Wales (NSW) and the Australian Capital Territory (ACT), and to explore the types of actions taken in response. Methods A cross-sectional survey of junior doctors in PGY1 or PGY2 positions was undertaken in 2015 and 2016 (n=374 and 440 respectively). Thematic analysis was undertaken on free-text responses to describe the reporting process and outcomes in more depth. Results The estimated response rate was 17-20%. Results from both surveys followed almost identical trends. Most respondents in 2015 and 2016 reported being bullied (n=203 (54.3%) and 253 (57.5%) respectively), 16-19% reported sexual harassment (n=58 and 82 respectively) and 29% of females reported sexual harassment. Qualitative analysis elucidated reasons for not taking action in response to bullying and harassment, including workplace normalisation of these behaviours, fear of reprisal and lack of knowledge or confidence in the reporting process. For respondents who did take action, most reported ineffective or personally harmful outcomes when reporting to senior colleagues, including being dismissed or blamed, and an intention not to trust the process in the future. Conclusions The findings suggest that interventions targeted at the level of junior doctors to improve the culture of bullying and harassment in medicine are unlikely to be helpful. Different approaches that address the problem in a more systemic way are needed, as is further research about the effectiveness of such interventions. What is known about the topic? Bullying and sexual harassment are common workplace experiences in the medical profession. What does this paper add? Over half the junior doctors in the present study experienced bullying and nearly one-fifth experienced sexual harassment. Junior doctors are reluctant to speak out, not only for fear of reprisal, but also because they do not believe it is worth doing so. What are the implications for practitioners? The data confirm a systemic problem of bullying in NSW. Primarily focusing on interventions with junior doctors (e.g. resilience training) is unlikely to solve the problem. Different and multipronged approaches (e.g. raising awareness in senior colleagues and training bystanders to intervene) should be tried and studied.


Possible mechanisms of cholesterol elevation aggravating COVID-19.
Importance: Despite the availability of a vaccine against the severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2), humans will have to live with this virus and the after-effects of the coronavirus disease 2019 (COVID-19) infection for a long time. Cholesterol plays an important role in the infection and prognosis of SARS-CoV-2, and the study of its mechanism is of great significance not only for the treatment of COVID-19 but also for research on generic antiviral drugs. Observations: Cholesterol promotes the development of atherosclerosis by activating NLR family pyrin domain containing 3 (NLRP3), and the resulting inflammatory environment indirectly contributes to COVID-19 infection and subsequent deterioration. In in vitro studies, membrane cholesterol increased the number of viral entry sites on the host cell membrane and the number of angiotensin-converting enzyme 2 (ACE2) receptors in the membrane fusion site. Previous studies have shown that the fusion protein of the virus interacts with cholesterol, and the spike protein of SARS-CoV-2 also requires cholesterol to enter the host cells. Cholesterol in blood interacts with the spike protein to promote the entry of spike cells, wherein the scavenger receptor class B type 1 (SR-B1) plays an important role. Because of the cardiovascular protective effects of lipid-lowering therapy and the additional anti-inflammatory effects of lipid-lowering drugs, it is currently recommended to continue lipid-lowering therapy for patients with COVID-19, but the safety of extremely low LDL-C is questionable. Conclusions and Relevance: Cholesterol can indirectly increase the susceptibility of patients to SARS-CoV-2 and increase the risk of death from COVID-19, which are mediated by NLRP3 and atherosclerotic plaques, respectively. Cholesterol present in the host cell membrane, virus, and blood may also directly participate in the virus cell entry process, but the specific mechanism still needs further study. Patients with COVID-19 are recommended to continue lipid-lowering therapy.


Mortality of late-preterm (near-term) newborns in Utah.
The purpose of this work was to determine the relative risk for mortality and the causes and ages of death for late-preterm newborns (gestational age of 34-36 weeks) compared with those born at term. We reviewed data from birth and death certificates of infants born in Utah between 1999 and 2004. We calculated early neonatal (first week), neonatal (first 28 days), and infant (first year) mortality rates for each weekly estimated gestational age cohort from 34 to 42 weeks and, using 40 weeks as the reference, risk ratios for each cohort. Causes of death were grouped into 8 categories and compared for near term and term newborns. Crude mortality rates and risk ratios for death from all causes and for infants dying of all causes other than birth defects were measured. Birth defects were the single-most common cause of death for both term and late-preterm newborns. Mortality rates for late-preterm newborns remained significantly higher after excluding those who died of birth defects from the comparisons. Compared with those born at term, late-preterm (near-term) newborns have significantly higher mortality rates. Each weekly increase in estimated gestational age is associated with a decreasing risk of death. Birth defects are the leading cause of death among late-preterm newborns but do not entirely account for their higher risk of death.