PAI-1 Regulates the Cytoskeleton and Intrinsic Stiffness of Vascular Smooth Muscle Cells.

BACKGROUND: Plasma concentration of PAI-1 (plasminogen activator inhibitor-1) correlates with arterial stiffness. Vascular smooth muscle cells (SMCs) express PAI-1, and the intrinsic stiffness of SMCs is a major determinant of total arterial stiffness. We hypothesized that PAI-1 promotes SMC stiffness by regulating the cytoskeleton and that pharmacological inhibition of PAI-1 decreases SMC and aortic stiffness. METHODS: PAI-039, a specific inhibitor of PAI-1, and small interfering RNA were used to inhibit PAI-1 expression in cultured human SMCs. Effects of PAI-1 inhibition on SMC stiffness, F-actin (filamentous actin) content, and cytoskeleton-modulating enzymes were assessed. WT (wild-type) and PAI-1-deficient murine SMCs were used to determine PAI-039 specificity. RNA sequencing was performed to determine the effects of PAI-039 on SMC gene expression. In vivo effects of PAI-039 were assessed by aortic pulse wave velocity. RESULTS: PAI-039 significantly reduced intrinsic stiffness of human SMCs, which was accompanied by a significant decrease in cytoplasmic F-actin content. PAI-1 gene knockdown also decreased cytoplasmic F-actin. PAI-1 inhibition significantly increased the activity of cofilin, an F-actin depolymerase, in WT murine SMCs, but not in PAI-1-deficient SMCs. RNA-sequencing analysis suggested that PAI-039 upregulates AMPK (AMP-activated protein kinase) signaling in SMCs, which was confirmed by Western blotting. Inhibition of AMPK prevented activation of cofilin by PAI-039. In mice, PAI-039 significantly decreased aortic stiffness and tunica media F-actin content without altering the elastin or collagen content. CONCLUSIONS: PAI-039 decreases intrinsic SMC stiffness and cytoplasmic stress fiber content. These effects are mediated by AMPK-dependent activation of cofilin. PAI-039 also decreases aortic stiffness in vivo. These findings suggest that PAI-1 is an important regulator of the SMC cytoskeleton and that pharmacological inhibition of PAI-1 has the potential to prevent and treat cardiovascular diseases involving arterial stiffening.

Study of Salmonella spp. from Cage Papers Belonging to Pet Birds in an Argentinean Canary Breeder Championship.

Birds, including canaries and other birds, have become increasingly popular as pets. Bird fairs, where breeders gather and show their production in a championship setting, present a setting for possible Salmonella spp. contamination and transmission. Therefore, this study estimated the rate of Salmonella spp. isolation from cage papers, located in the bottom of cages of exotic pet birds, including canaries. Collected Salmonella isolates were used to determine the antimicrobial resistance profile to 52 antibiotics and 17 commercial disinfectants, based on pure or a mixture of acids, alcohols, aldehydes, alkalis, halogens, peroxygen, and quaternary ammonium compounds. The samples consisted of 774 cage papers taken in the 2015 Argentinean canary breeder championship, pooling three cage papers into one sterile sampling bag. Only one pool of the cage papers was positive for Salmonella spp. (0.4%), which belonged to the sample from three frill canary cages. Two strains of Salmonella serotype Glostrup were isolated, which were only resistant to sulfonamides and erythromycin and sensitive to alkali-based product PL301 AS. Although the rate of Salmonella spp. isolation from cage papers in an Argentinean canary breeder championship is low, it should not be discounted because Salmonella ser. Glostrup can be a source of human Salmonella outbreaks and they show high resistance to disinfecting products.

Hepatopancreatic metabolic disorders and their implications in the development of Alzheimer's disease and vascular dementia.

Dementia has been faced with significant public health challenges and economic burdens that urges the need to develop safe and effective interventions. In recent years, an increasing number of studies have focused on the relationship between dementia and liver and pancreatic metabolic disorders that result in diseases such as diabetes, obesity, hypertension and dyslipidemia. Previous reports have shown that there is a plausible correlation between pathologies caused by hepatopancreatic dysfunctions and dementia. Glucose, insulin and IGF-1 metabolized in the liver and pancreas probably have an important influence on the pathophysiology of the most common dementias: Alzheimer's and vascular dementia. This current review highlights recent studies aimed at identifying convergent mechanisms, such as insulin resistance and other diseases, linked to altered hepatic and pancreatic metabolism, which are capable of causing brain changes that ultimately lead to dementia.

Neuraminidase-induced externalization of phosphatidylserine activates ADAM17 and impairs insulin signaling in endothelial cells.

Endothelial insulin resistance represents a causal factor in the pathogenesis of type 2 diabetes (T2D) and vascular disease, thus the need to identify molecular mechanisms underlying defects in endothelial insulin signaling. We previously have shown that a disintegrin and metalloproteinase-17 (ADAM17) is increased while insulin receptor α-subunit (IRα) is decreased in the vasculature of patients with T2D, leading to impaired insulin-induced vasodilation. We have also demonstrated that ADAM17 sheddase activity targets IRα; however, the mechanisms driving endothelial ADAM17 activity in T2D are largely unknown. Herein, we report that externalization of phosphatidylserine (PS) to the outer leaflet of the plasma membrane causes ADAM17-mediated shedding of IRα and blunting of insulin signaling in endothelial cells. Furthermore, we demonstrate that endothelial PS externalization is mediated by the phospholipid scramblase anoctamin-6 (ANO6) and that this process can be stimulated by neuraminidase, a soluble enzyme that cleaves sialic acid residues. Of note, we demonstrate that men and women with T2D display increased levels of neuraminidase activity in plasma, relative to age-matched healthy individuals, and this occurs in conjunction with increased ADAM17 activity and impaired leg blood flow responses to endogenous insulin. Collectively, this work reveals the neuraminidase-ANO6-ADAM17 axis as a novel potential target for restoring endothelial insulin sensitivity in T2D.NEW & NOTEWORTHY This work provides the first evidence that neuraminidase, an enzyme increased in the circulation of men and women with type 2 diabetes (T2D), promotes anoctamin-6 (ANO6)-dependent externalization of phosphatidylserine in endothelial cells, which in turn leads to activation of a disintegrin and metalloproteinase-17 (ADAM17) and consequent shedding of the insulin receptor-α from the cell surface. Hence, this work supports that consideration should be given to the neuraminidase-ANO6-ADAM17 axis as a novel potential target for restoring endothelial insulin sensitivity in T2D.

Neuraminidase inhibition improves endothelial function in diabetic mice.

Neuraminidases cleave sialic acids from glycocalyx structures and plasma neuraminidase activity is elevated in type 2 diabetes (T2D). Therefore, we hypothesize circulating neuraminidase degrades the endothelial glycocalyx and diminishes flow-mediated dilation (FMD), whereas its inhibition restores shear mechanosensation and endothelial function in T2D settings. We found that compared with controls, subjects with T2D have higher plasma neuraminidase activity, reduced plasma nitrite concentrations, and diminished FMD. Ex vivo and in vivo neuraminidase exposure diminished FMD and reduced endothelial glycocalyx presence in mouse arteries. In cultured endothelial cells, neuraminidase reduced glycocalyx coverage. Inhalation of the neuraminidase inhibitor, zanamivir, reduced plasma neuraminidase activity, enhanced endothelial glycocalyx length, and improved FMD in diabetic mice. In humans, a single-arm trial (NCT04867707) of zanamivir inhalation did not reduce plasma neuraminidase activity, improved glycocalyx length, or enhanced FMD. Although zanamivir plasma concentrations in mice reached 225.8 ± 22.0 ng/mL, in humans were only 40.0 ± 7.2 ng/mL. These results highlight the potential of neuraminidase inhibition for ameliorating endothelial dysfunction in T2D and suggest the current Food and Drug Administration-approved inhaled dosage of zanamivir is insufficient to achieve desired outcomes in humans.NEW & NOTEWORTHY This work identifies neuraminidase as a key mediator of endothelial dysfunction in type 2 diabetes that may serve as a biomarker for impaired endothelial function and predictive of development and progression of cardiovascular pathologies associated with type 2 diabetes (T2D). Data show that intervention with the neuraminidase inhibitor zanamivir at effective plasma concentrations may represent a novel pharmacological strategy for restoring the glycocalyx and ameliorating endothelial dysfunction.

Leaf carbon isotope tracks the facilitation pattern of legume shrubs shaped by water availability and species replacement along a large elevation gradient in Trans-Himalayas.

BACKGROUND AND AIMS: Understanding patterns and mechanisms of nurse plant facilitation is important to predict the resilience of arid/semi-arid ecosystems to climate change. We investigate whether water availability and nurse species turnover interact to shape the facilitation pattern of widespread legume shrubs along a large elevation gradient. We also investigate whether leaf δ13C of nurse plants can track the facilitation pattern. METHODS: We measured the relative interaction index (RII) of the number of species within and outside the canopy of two widespread legume shrub species (Caragana gerardiana and Caragana versicolor) alternatively distributed along a large elevation gradient in the Trans-Himalayas. We also assessed the proportional increase of species richness (ISR) at the community level using the paired plot data. To determine site-specific water availability, we measured the leaf δ13C of nurse shrubs and calculated the Thornthwaite moisture index (MI) for each elevation site. KEY RESULTS: Elevational variations in RII, ISR and δ13C were mainly explained by the MI when the effects of soil nitrogen and plant traits (leaf nitrogen and shrub size) were controlled. Variations in RII and ISR across the two nurse species were explained better by δ13C than by smoothly changing climatic factors along elevation. At the transition zone between the upper limit of C. gerardiana (4100 m) and the lower limit of C. versicolor (4200 m), RII and ISR were much higher in C. versicolor than in C. gerardiana under a similar MI. Such an abrupt increase in facilitation induced by nurse species replacement was well tracked by the variation of δ13C. CONCLUSIONS: Water availability and nurse species replacement are crucial to shaping facilitation patterns by legume shrubs along a large elevation gradient in dry mountainous regions, such as the Trans-Himalayas. Turnover in nurse species under global change might significantly alter the pattern of nurse plant facilitation associated with water availability, which can be well tracked by leaf δ13C.

Prevalence of alcohol use during pregnancy, Brazil, 2011-2012. / Prevalência de uso de álcool na gestação, Brasil, 2011-2012.

This is a national cross-sectional, hospital-based study, which interviewed 23,894 postpartum women in 2011-2012 aiming to estimate the prevalence of alcohol use during pregnancy and identifying more vulnerable groups. Alcohol use during pregnancy was identified using the TWEAK scale, and women with a score of ≥ 2 were classified as having a "presumable diagnosis of inadequate alcohol use". The national prevalence of alcohol use and the prevalence in subgroups were calculated according to maternal characteristics, with respective 95% confidence intervals (95%CI). Coexistence of smoking, inadequate prenatal consultations, and alcohol use during pregnancy were graphically identified. The prevalence of alcohol use was 14% (95%CI: 13.3-14.7), with 10% (95%CI: 9.3-10.6) of women presenting presumable diagnosis of inadequate alcohol us during pregnancy. Higher prevalence of alcohol use and presumable diagnosis of inadequate alcohol us was observed in black women, aged 12-19 years, with lower educational level, from a lower economic class, without a partner, without paid work, with more than three previous births, who did not want to get pregnant, with inadequate prenatal care, with previous delivery in public services, and who reported smoking during pregnancy. Among the interviewees, 1.2% presented all three risk factors for negative perinatal outcomes at the same time: smoking, alcohol use, and inadequate prenatal care. The results showed a high prevalence of alcohol use during pregnancy and presumable diagnosis of inadequate alcohol us, especially among women with worse social conditions. These data are relevant for the formulation of public policies to prevent alcohol use and provide support services to help this population stop alcohol use during pregnancy.

Estudo transversal, de base hospitalar, nacional, com entrevista de 23.894 puérperas, em 2011-2012, com os objetivos de estimar a prevalência de consumo de álcool na gestação e identificar grupos mais vulneráveis. O uso de álcool na gestação foi identificado por meio da escala TWEAK, sendo classificadas como "diagnóstico presumível de uso inadequado de álcool" mulheres com pontuação ≥ 2. Calculou-se a prevalência nacional de uso de álcool e em subgrupos de acordo com características maternas, com respectivos intervalos de 95% de confiança (IC95%). Foram encontradas, de forma gráfica, coexistência de tabagismo, inadequação de consultas pré-natais e ingestão de bebidas alcoólicas na gestação. A prevalência de uso de álcool foi de 14% (IC95%: 13,3-14,7), com 10% (IC95%: 9,3-10,6) das mulheres apresentando diagnóstico presumível de uso inadequado de álcool na gestação. Maiores prevalências de uso de álcool e de diagnóstico presumível de uso inadequado foram observadas em mulheres pretas, com 12-19 anos de idade, com menor índice de escolaridade, de classe econômica mais baixa, sem companheiro, sem trabalho remunerado, com mais de três partos anteriores, que não queriam engravidar, com assistência pré-natal inadequada, com parto em serviços públicos e que referiram tabagismo na gestação. Estima-se que 1,2% das mulheres entrevistadas apresentavam concomitância dos três fatores de risco para desfechos perinatais negativos: fumo, álcool e assistência pré-natal inadequada. Os resultados demonstraram alta prevalência de uso de álcool na gestação e de diagnóstico presumível de uso inadequado, principalmente por mulheres em situação de vulnerabilidade social. São relevantes a elaboração de políticas públicas que contemplem ações de prevenção do uso de bebidas alcoólicas e a prestação de serviços de apoio para cessação do uso de álcool na gravidez.

Estudio transversal, de base hospitalaria, nacional, con entrevistas a 23.894 puérperas, en 2011-2012, con el objetivo de estimar la prevalencia de consumo de alcohol durante el embarazo e identificar grupos más vulnerables. El consumo de alcohol durante el embarazo se identificó mediante la escala TWEAK, y las mujeres con puntuación ≥ 2 fueron clasificadas como "diagnóstico presumible de uso inadecuado de alcohol¨. Se calculó la prevalencia nacional de consumo de alcohol y subgrupos según características maternas, con sus respectivos intervalos de 95% de confianza (IC95%). Se identificó gráficamente la coexistencia de tabaquismo, consultas prenatales inadecuadas y consumo de alcohol durante el embarazo. La prevalencia de consumo de alcohol fue del 14% (IC95%: 13,3-14,7), siendo el 10% (IC95%: 9,3-10,6) de mujeres con diagnóstico presumible de uso inadecuado durante el embarazo. Se observaron mayores prevalencias de consumo de alcohol y diagnóstico presumible de uso inadecuado en mujeres de color/raza negra, de 12-19 años, con menos años de escolaridad, de clase económica más baja, sin pareja, sin trabajo remunerado, con más de tres partos previos, que no querían quedar embarazadas, con control prenatal inadecuado, con parto en el sistema público de salud y que relataron fumar durante el embarazo. Se estima que el 1,2% de las mujeres entrevistadas presentaron la concomitancia de tres factores de riesgo en resultados perinatales negativos: tabaquismo, alcohol y atención prenatal inadecuada. Los resultados demuestran una alta prevalencia de consumo de alcohol durante el embarazo y de diagnóstico presumible de uso inadecuado, especialmente en mujeres con las peores condiciones sociales, siendo relevante para la elaboración de políticas públicas que incluyan acciones de prevención del consumo de alcohol y servicios de apoyo para el cese del consumo de alcohol en el embarazo.

Physical violence against women in Brazil: Findings from the 3rd Brazilian household survey on substance use.

Brazil has a historical gap regarding information on violence against women. Herein we aimed to evaluate the association of socioeconomic and demographic characteristics with physical violence against women in Brazil, as well as the possible escalation of violence to severe patterns of violence. We analysed data from the 3rd Brazilian Household Survey on Substance Use, in 2015. The main outcomes were reporting any physical violence and being stabbed/shot in the last 12-months. Logistic regressions were fitted to assess the association between socioeconomic and demographic variables with the outcomes. We estimated 3.8 million women reported any physical violence (5.52%): 3.79% reported threats to beat/ push/kick, 1.87% threats with knife/gun, 2.49% were beaten/pushed/kicked, 0.63% were spanked/ choked, and 0.21% were stabbed/shot. The higher the severity of violence, the higher the number of types of violence experienced. The likelihood of reporting any violence was higher among women 18-24 years, without a stable partner, who were at an informal job or unemployed, and who live in urban areas. The sociodemographic characteristics associated with reporting any violence reinforce the importance of addressing gender inequalities. Evidence of escalation violence reinforces the need to protect and care for women who report any type of violence.

Detección cualitativa de sulfitos en productos cárnicos comercializados en la provincia de Santa Fe, Argentina / Qualitative detection of sulfites in meat products marketed in the province of Santa Fe, Argentina

Los sulfitos son aditivos que se utilizan como conservantes y antioxidantes en ciertos alimentos. De acuerdo a lo establecido en el Código Alimentario Argentino, no está permitido su uso en carnes crudas debido a que enmascara el verdadero estado de conservación y, al descomponer la vitamina B1, disminuye su valor nutricional. Por ello, el objetivo de este trabajo fue detectar cualitativamente la presencia de agentes sulfitantes en productos cárnicos frescos de carnicerías de diferentes localidades de la provincia de Santa Fe, con el fin de fiscalizar el uso de este aditivo. Las muestras fueron tomadas a granel y recibidas en el laboratorio de la Agencia Santafesina de Seguridad Alimentaria a temperatura de refrigeración. En el período comprendido entre los años 2011 y 2018, ingresaron un total de 1071 muestras de carnes y preparados de carnes frescas, de las cuales 532 fueron de carne picada, 357 de chorizo fresco, 149 de hamburguesa y 33 de albóndiga. En primer lugar, se realizó un screening con la técnica del verde de malaquita (AOAC 961.09) y, con fines confirmatorios, las muestras que resultaron positivas fueron analizadas cualitativamente con el método de Monier-Williams optimizado (AOAC 990.28). El estudio integral demostró que 28 muestras de carne picada (5,3 %), 59 de chorizo fresco (16,0 %), 25 de hamburguesa (16,7 %) y 2 de albóndiga (6,1 %) contenían este aditivo. En conclusión, según los datos obtenidos, el principal uso de las sales de sulfitos en algunas industrias cárnicas se dio en la elaboración de chorizos frescos y hamburguesas; por ello, el control de estos alimentos es una tarea fundamental para los entes fiscalizadores.

Sulfites are additives that are used as preservatives and antioxidants in certain foods. According to the Argentine Food Code (CAA), their use is not allowed in raw meats because it masks the true state of preservation and diminishes their nutritional value by breaking down vitamin B1. Therefore, the objective of this work was to qualitatively detect the presence of sulfiting agents in fresh meat products from butcher shops in different locations in the province of Santa Fe, in order to control the use of this additive. The samples were taken in bulk and received in the laboratory of the Santa Fe Food Safety Agency (ASSAl) at refrigeration temperature. Between 2011 and 2018, a total of 1,071 samples of meat and fresh meat products were submitted, including minced meat (532), fresh sausages (357), hamburgers (149), and meatballs (33). First, a screening was conducted with the malachite green technique (AOAC 961.09), and for confirmatory purposes, the samples that tested positive were qualitatively analyzed using the optimized Monier-Williams method (AOAC 990.28). The comprehensive study showed that 28 samples of minced meat (5.3 %), 59 samples of fresh sausage (16.0 %), 25 samples of hamburger (16.7 %), and 2 samples of meatballs (6.1 %) contained this additive. In conclusion, based on the data obtained, the main use of sulfite salts in some meat industries was found in the production of fresh sausages and hamburgers; hence, the control of these foods is a crucial task for regulatory authorities.

Lifestyle changes during the COVID-19 pandemic in Brazil: results from three consecutive cross-sectional web surveys.

OBJECTIVE: The importance of a healthy lifestyle in preventing morbidity and mortality is well-established. The COVID-19 pandemic brought about significant lifestyle changes globally, but the extent of these changes in the Brazilian population remains unclear. The objective of this study was to evaluate changes in lifestyle among the Brazilian general population during the first year of the pandemic. DESIGN: Three consecutive anonymous web surveys were carried out: survey 1 (S1)-April 2020, S2-August 2020 and S3-January 2021. SETTING: Brazil. PARTICIPANTS: The study included 19 257 (S1), 1590 (S2) and 859 (S3) participants from the general population, who were ≥18 years, of both sexes, with access to the internet, self-reporting living in Brazil and who agreed to participate after reading the informed consent. PRIMARY OUTCOME: Lifestyle changes were assessed using the Short Multidimensional Instrument for Lifestyle Evaluation-Confinement (SMILE-C). The SMILE-C assesses lifestyle across multiple domains including diet, substance use, physical activity, stress management, restorative sleep, social support and environmental exposures. We used a combination of bootstrapping and linear fixed-effect modelling to estimate pairwise mean differences of SMILE-C scores overall and by domain between surveys. RESULTS: In all the surveys, participants were mostly women and with a high education level. Mean SMILE-C scores were 186.4 (S1), 187.4 (S2) and 190.5 (S3), indicating a better lifestyle in S3 as compared with S1. The pairwise mean differences of the overall SMILE-C scores were statistically significant (p<0.001). We also observed a better lifestyle over time in all domains except for diet and social support. CONCLUSIONS: Our findings indicate that individuals from a large middle-income country, such as Brazil, struggled to restore diet and social relationships after 1 year of the pandemic. These findings have implications for monitoring the long-term consequences of the pandemic, as well as future pandemics.

Differences in Muscle Lipogenic Gene Expression, Carcass Traits and Fat Deposition among Three Iberian Pig Strains Finished in Two Different Feeding Systems.

The Iberian pig breed includes several well-differentiated strains. The present study evaluated carcass traits, fat deposition and muscle expression of important lipogenic genes (SCD, ME1, ACACA, FASN, EGR1, ACOX and ACLY) using 65 male pigs of 3 Iberian strains (20 Lampiño, 23 Torbiscal, and 22 Retinto) finished either in a conventional, concentrate-based system (CF) or in montanera (MF), a traditional free-range system with acorn feeding. Torbiscal had the highest ham, Longissimus thoracis and prime cuts yields, and the thinnest subcutaneous adipose tissue (SAT). Retinto had the highest monounsaturated fatty acids (MUFA) and percentage of intramuscular fat (IMF), while Lampiño had the greatest content of saturated fatty acids (SFA), polyunsaturated fatty acids (PUFA), atherogenic (AI) and thrombogenic (TI) indexes in SAT. Conventionally finished pigs had the highest ham, L. thoracis and prime cuts yields, and SFA. Montanera-finished animals had the highest PUFA and MUFA contents, and the lowest AI, TI and n6/n3 ratio in SAT. In relation to gene expression, Retinto had the greatest SCD, FASN and ACLY levels. Most studied genes were overexpressed in CF pigs. In conclusion, MF pigs had healthier fat than CF pigs, and Retinto had the healthiest fat and the greatest lipogenic trend in muscle, supported by IMF and lipogenic gene expression.

A historical, economic, and technical-scientific approach to the current crisis in the development of antibacterial drugs: Promising role of antibacterial peptides in this scenario.

The emergence of antibiotic resistance (AMR) is a global public health problem. According to estimates, drug-resistant bacteria infect 2 million patients and perish 23,000 annually. To overcome this problem, antimicrobial peptides became a potential solution based on a new mechanism of action against bacteria. This article addresses the phenomenon of antibacterial resistance in most of its nuances, responding to historical, technical-scientific, and economic aspects. Likewise, it explores new therapeutic approaches to combat multi-resistant pathogens, specifically concerning antibacterial peptides, as a potential therapeutic tool to mitigate the current crisis of antibacterial drugs. It is expected that, with technological advances, especially with the advent and adoption of artificial intelligence, there will be an increase in diversified synthetic peptide production, which can face the challenges that we have in terms of antibacterial drugs.

Giving a Hand: Synthetic Peptides Boost the Antifungal Activity of Itraconazole against Cryptococcus neoformans.

Cryptococcus neoformans is a multidrug-resistant pathogen responsible for infections in immunocompromised patients. Here, itraconazole (ITR), a commercial antifungal drug with low effectiveness against C. neoformans, was combined with different synthetic antimicrobial peptides (SAMPs), Mo-CBP3-PepII, RcAlb-PepII, RcAlb-PepIII, PepGAT, and PepKAA. The Mo-CBP3-PepII was designed based on the sequence of MoCBP3, purified from Moringa oleifera seeds. RcAlb-PepII and RcAlb-PepIII were designed using Rc-2S-Alb, purified from Ricinus communis seed cakes. The putative sequence of a chitinase from Arabidopsis thaliana was used to design PepGAT and PepKAA. All SAMPs have a positive liquid charge and a hydrophobic potential ranging from 41-65%. The mechanisms of action responsible for the combined effect were evaluated for the best combinations using fluorescence microscopy (FM). The synthetic peptides enhanced the activity of ITR by 10-fold against C. neoformans. Our results demonstrated that the combinations could induce pore formation in the membrane and the overaccumulation of ROS on C. neoformans cells. Our findings indicate that our peptides successfully potentialize the activity of ITR against C. neoformans. Therefore, synthetic peptides are potential molecules to assist antifungal agents in treating Cryptococcal infections.

No Chance to Survive: Mo-CBP3-PepII Synthetic Peptide Acts on Cryptococcus neoformans by Multiple Mechanisms of Action.

Multidrug-resistant Cryptococcus neoformans is an encapsulated yeast causing a high mortality rate in immunocompromised patients. Recently, the synthetic peptide Mo-CBP3-PepII emerged as a potent anticryptococcal molecule with an MIC50 at low concentration. Here, the mechanisms of action of Mo-CBP3-PepII were deeply analyzed to provide new information about how it led C. neoformans cells to death. Light and fluorescence microscopies, analysis of enzymatic activities, and proteomic analysis were employed to understand the effect of Mo-CBP3-PepII on C. neoformans cells. Light and fluorescence microscopies revealed Mo-CBP3-PepII induced the accumulation of anion superoxide and hydrogen peroxide in C. neoformans cells, in addition to a reduction in the activity of superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT) in the cells treated with Mo-CBP3-PepII. In the presence of ascorbic acid (AsA), no reactive oxygen species (ROS) were detected, and Mo-CBP3-PepII lost the inhibitory activity against C. neoformans. However, Mo-CBP3-PepII inhibited the activity of lactate dehydrogenase (LDH) ergosterol biosynthesis and induced the decoupling of cytochrome c (Cyt c) from the mitochondrial membrane. Proteomic analysis revealed a reduction in the abundance of proteins related to energetic metabolism, DNA and RNA metabolism, pathogenicity, protein metabolism, cytoskeleton, and cell wall organization and division. Our findings indicated that Mo-CBP3-PepII might have multiple mechanisms of action against C. neoformans cells, mitigating the development of resistance and thus being a potent molecule to be employed in the production of new drugs against C. neoformans infections.

Shrub-mediated effects on soil nitrogen determines shrub-herbaceous interactions in drylands of the Tibetan Plateau.

Introduction: Shrub promotes the survival, growth and reproduction of understory species by buffering the environmental extremes and improving limited resources (i.e., facilitation effect) in arid and semiarid regions. However, the importance of soil water and nutrient availability on shrub facilitation, and its trend along a drought gradient have been relatively less addressed in water-limited systems. Methods: We investigated species richness, plant size, soil total nitrogen and dominant grass leaf δ13C within and outside the dominant leguminous cushion-like shrub Caragana versicolor along a water deficit gradient in drylands of Tibetan Plateau. Results: We found that C. versicolor increased grass species richness but had a negative effect on annual and perennial forbs. Along the water deficit gradient, plant interaction assessed by species richness (RIIspecies) showed a unimodal pattern with shift from increase to decrease, while plant interaction assessed by plant size (RIIsize) did not vary significantly. The effect of C. versicolor on soil nitrogen, rather than water availability, determined its overall effect on understory species richness. Neither the effect of C. versicolor on soil nitrogen nor water availability affected plant size. Discussion: Our study suggests that the drying tendency in association with the recent warming trends observed in drylands of Tibetan Plateau, will likely hinder the facilitation effect of nurse leguminous shrub on understories if moisture availability crosses a critical minimum threshold.

Impact of sex and diet-induced weight loss on vascular insulin sensitivity in type 2 diabetes.

Vascular insulin resistance, a major characteristic of obesity and type 2 diabetes (T2D), manifests with blunting of insulin-induced vasodilation. Although there is evidence that females are more whole body insulin sensitive than males in the healthy state, whether sex differences exist in vascular insulin sensitivity is unclear. Also uncertain is whether weight loss can reestablish vascular insulin sensitivity in T2D. The purpose of this investigation was to 1) establish if sex differences in vasodilatory responses to insulin exist in absence of disease, 2) determine whether female sex affords protection against the development of vascular insulin resistance with long-term overnutrition and obesity, and 3) examine if diet-induced weight loss can restore vascular insulin sensitivity in men and women with T2D. First, we show in healthy mice and humans that sex does not influence insulin-induced femoral artery dilation and insulin-stimulated leg blood flow, respectively. Second, we provide evidence that female mice are protected against impairments in insulin-induced dilation caused by overnutrition-induced obesity. Third, we show that men and women exhibit comparable levels of vascular insulin resistance when T2D develops but that diet-induced weight loss is effective at improving insulin-stimulated leg blood flow, particularly in women. Finally, we provide indirect evidence that these beneficial effects of weight loss may be mediated by a reduction in endothelin-1. In aggregate, the present data indicate that female sex confers protection against obesity-induced vascular insulin resistance and provide supportive evidence that, in women with T2D, vascular insulin resistance can be remediated with diet-induced weight loss.

The relaxation zone: Initial analysis of stress management services for university students.

Objective: We evaluated the Relaxation Zone (RZ), a room with "relaxation stations" designed to reduce stress in college students. Participants: Participants (N = 994) were enrolled during the 2019 calendar year. Most participants were female (62.8%), freshmen (59.3%), and African American (44.9%). Methods: Students visiting the RZ completed a brief pretest and posttest that measured self-reported stress. Results: Paired sample t-tests across class groups (e.g., freshman, sophomore) revealed a significant reduction in stress from pretest (Mpooled = 6.0) to posttest (Mpooled = 4.5), with medium to large effect sizes (Cohen's d ranging from 0.53 to 0.88) during their first visit. For students having at least two visits (N = 348), an exploratory dose-response analysis demonstrated that an optimal dose of self-guided relaxation ranged from two to eight visits. Conclusions: These findings provide initial evidence that programs like the RZ can reduce student stress within a few visits.

Functional Implications and Clinical Potential of MicroRNAs in Irritable Bowel Syndrome: A Concise Review.

MicroRNAs (miRNAs) are tiny (20-24 nucleotides long), non-coding, highly conserved RNA molecules that play a crucial role within the post-transcriptional regulation of gene expression via sequence-specific mechanisms. Since the miRNA transcriptome is involved in multiple molecular processes needed for cellular homeostasis, its altered expression can trigger the development and progression of several human pathologies. In this context, over the last few years, several relevant studies have demonstrated that dysregulated miRNAs affect a wide range of molecular mechanisms associated with irritable bowel syndrome (IBS), a common gastrointestinal disorder. For instance, abnormal miRNA expression in IBS patients is related to the alteration of intestinal permeability, visceral hyperalgesia, inflammatory pathways, and pain sensitivity. Besides, specific miRNAs are differentially expressed in the different subtypes of IBS, and therefore, they might be used as biomarkers for precise diagnosis of these pathological conditions. Accordingly, miRNAs have noteworthy potential as theragnostic targets for IBS. Hence, in this current review, we present an overview of the recent discoveries regarding the clinical relevance of miRNAs in IBS, which might be useful in the future for the development of miRNA-based drugs against this disorder.