The evolutionary conserved miR-137/325 tandem mediates obesity-induced hypogonadism and metabolic comorbidities by repressing hypothalamic kisspeptin.

BACKGROUND: Obesity-induced hypogonadism (OIH) is a prevalent, but often neglected condition in men, which aggravates the metabolic complications of overweight. While hypothalamic suppression of Kiss1-encoded kisspeptin has been suggested to contribute to OIH, the molecular mechanisms for such repression in obesity, and the therapeutic implications thereof, remain unknown. METHODS: A combination of bioinformatic, expression and functional analyses was implemented, assessing the role of the evolutionary-conserved miRNAs, miR-137 and miR-325, in mediating obesity-induced suppression of hypothalamic kisspeptin, as putative mechanism of central hypogonadism and metabolic comorbidities. The implications of such miR-137/325-kisspeptin interplay for therapeutic intervention in obesity were also explored using preclinical OIH models. RESULTS: MiR-137/325 repressed human KISS1 3'-UTR in-vitro and inhibited hypothalamic kisspeptin content in male rats, while miR-137/325 expression was up-regulated, and Kiss1/kisspeptin decreased, in the medio-basal hypothalamus of obese rats. Selective over-expression of miR-137 in Kiss1 neurons reduced Kiss1/ kisspeptin and partially replicated reproductive and metabolic alterations of OIH in lean mice. Conversely, interference of the repressive actions of miR-137/325 selectively on Kiss1 3'-UTR in vivo, using target-site blockers (TSB), enhanced kisspeptin content and reversed central hypogonadism in obese rats, together with improvement of glucose intolerance, insulin resistance and cardiovascular and inflammatory markers, despite persistent exposure to obesogenic diet. Reversal of OIH by TSB miR-137/325 was more effective than chronic kisspeptin or testosterone treatments in obese rats. CONCLUSIONS: Our data disclose that the miR-137/325-Kisspeptin repressive interaction is a major player in the pathogenesis of obesity-induced hypogonadism and a putative druggable target for improved management of this condition and its metabolic comorbidities in men suffering obesity. SIGNIFICANCE STATEMENT: Up to half of the men suffering obesity display also central hypogonadism, an often neglected complication of overweight that can aggravate the clinical course of obesity and its complications. The mechanisms for such obesity-induced hypogonadism remain poorly defined. We show here that the evolutionary conserved miR137/miR325 tandem centrally mediates obesity-induced hypogonadism via repression of the reproductive-stimulatory signal, kisspeptin; this may represent an amenable druggable target for improved management of hypogonadism and other metabolic complications of obesity.

Does facial structure explain differences in student evaluations of teaching? The role of fWHR as a proxy for perceived dominance.

Dominance is usually viewed as a positive male attribute, but this is not typically the case for women. Using a novel dataset of student evaluations of teaching in a school of Business and Economics of a selective university, we construct the face width-to-height ratio (fWHR) as a proxy for perceived dominance to assess whether individuals with a higher ratio obtain better evaluations. Our results show that a higher fWHR is associated with a better evaluation for male faculty, while the opposite is the case for female faculty. These results are not due to differences in teachers' quality or beauty. In terms of magnitude, the effect of the fWHR is much larger for female professors. To the extent that fWHR is a good proxy of perceived dominance, it appears that conformity to traditional gender norms pays off for both men and women. However, the cost of challenging these norms is much larger for women than for men.

Status of head and neck brachytherapy in Spain in 2022.

PURPOSE: Brachytherapy (BT) has been used for many years for disease control in tumours of the head and neck area (H&N). It is currently performed with high dose rate (HDR) or pulsed dose rate (PDR), but its use has been reduced due to the implementation of new non-invasive external beam radiotherapy techniques such as intensity modulation (IMRT) and volumetric modulated arc therapy (VMAT) and the improvement of surgical techniques. METHODS: The Spanish Brachytherapy Group (GEB) has carried out a survey to find out the number of centres in Spain that continue to use BT in H&N and its indications and expectations for the future. RESULTS: The results were presented at the XX GEB Consensus Meeting held on October 21, 2022, in Valencia (Spain) and it was confirmed that, although there are fewer and fewer centres that use BT in H&N, there are still units with extensive experience in this technique that should be positioned as referral centres. CONCLUSION: It is necessary to carry out continuous work with other specialities involved, such as H&N surgeons, and other radiation oncologists, to improve the training of residents, both oncologists and medical physicists.

Reflexiones sobre el desentrenamiento deportivo como competencia profesional específica

El desentrenamiento deportivo, como competencia profesional específica, constituye objeto de trabajo en la formación inicial del Licenciado en Cultura Física, al ser este un actor social en contextos sociales y desde esta perspectiva, se ha orientado la mirada reflexiva sobre su importancia; se precisa además, como una competencia que posibilita la asesoría para la salud desde el eslabón de base no solo en el alto rendimiento, de ahí la necesidad de desarrollarla con un enfoque salubrista, preventivo y profiláctico. Aunque varios estudios abordan esta temática, se detectaron carencias teóricas en los modelos de referencia; en consecuencia, se trazó como objetivo del trabajo reflexionar sobre el desentrenamiento deportivo como competencia profesional específica en la formación del Licenciado en Cultura Física, desde la actividad laboral investigativa. Se emplearon métodos como el análisis-síntesis y el sistémico estructural funcional que posibilitaron profundizar en investigaciones sobre las competencias profesionales con una óptica analítica, interpretativa y crítica. Como conclusiones, se corroboraron las limitaciones teóricas relacionadas con el desentrenamiento deportivo como competencia profesional específica, así como la posibilidad de complementar y enriquecer el modelo actuante, a partir de la práctica formativa desde el pregrado, lo que contribuyó a brindar mejor solución a los problemas del desentrenamiento en el eslabón de base.

A desformação desportiva, enquanto competência profissional específica, constitui objeto de trabalho na formação inicial do Licenciado em Cultura Física, por ser este um ator social em contextos sociais e nesta perspetiva tem orientado o olhar reflexivo sobre a sua importância; Exige-se também como uma competência que possibilite o aconselhamento em saúde desde a base, não apenas no alto rendimento, daí a necessidade de desenvolvê-la com enfoque sanitário, preventivo e profilático. Embora vários estudos abordem essa questão, foram detectadas deficiências teóricas nos modelos de referência; Diante disso, o objetivo do trabalho foi refletir sobre o destreinamento esportivo como competência profissional específica na formação do Bacharel em Cultura Física, a partir da atividade laboral investigativa. Foram utilizados métodos como análise-síntese e sistêmico-funcional estrutural, que possibilitaram o aprofundamento da pesquisa sobre competências profissionais a partir de uma perspectiva analítica, interpretativa e crítica. Como conclusões, foram corroboradas as limitações teóricas relacionadas à destreinamento esportivo como competência profissional específica, bem como a possibilidade de complementar e enriquecer o modelo de atuação, baseado na prática de treinamento desde a graduação, o que contribuiu para uma melhor solução dos problemas. o elo básico.

Detraining in sports, as a specific professional competence, constitutes an object of work in the initial training of the Bachelor of Physical Culture degree, as this is a social actor in social contexts and from this perspective, the reflective gaze on its importance has been oriented; it is also required as a competence that enables health advice from the base link, not only in high performance, hence the need to develop it with a health, preventive and prophylactic approach. Although several studies address this issue, theoretical deficiencies were detected in the reference models; consequently, the objective of the work was to reflect on sports detraining as a specific professional competence in the formation of the Bachelor of Physical Culture degree, from the research work activity. Methods such as analysis-synthesis and structural-functional systemic were used, which made it possible to deepen research on professional skills from an analytical, interpretative and critical perspective. As conclusions, the theoretical limitations related to sports detraining as a specific professional competence were corroborated, as well as the possibility of complementing and enriching the acting model, based on training practice from undergraduate, which contributed to providing a better solution to problems. detraining in the base link.

Seroprevalence of SARS-CoV-2 antibodies in schoolchildren in the city of São Paulo, 2020.

OBJECTIVE: To estimate seroprevalence of SARS-CoV-2 antibodies in schoolchildren aged 4 to 14 years living in the city of São Paulo, according to clinical, demographic, epidemiological, and social variables, during the school closure period as a measure against covid-19 spread. METHODS: A serological survey was made in September 2020 with a random sample stratified by school system (municipal public, state public and private) type. A venous blood sample was collected using the Wondfo SARS-CoV-2 Antibody Test (lateral flow method) for detection of total SARS-CoV-2 virus antibodies. Semi-structured questionnaires were applied to collect clinical, demographic, social, and epidemiological data. RESULTS: Seroprevalence of SARS-CoV-2 antibodies in schoolchildren was of 16.6% (95%CI 15.4-17.8). The study found higher seroprevalence in the municipal (18.5%; 95%CI 16.6-20.6) and state (16.2%; 95%CI 14.4-18.2) public school systems compared to the private school system (11.7; 95%CI 10.0-13.7), among black and brown students (18.4%; 95%CI 16.8-20.2) and in the most vulnerable social stratum (18.5 %;95%CI 16.9-20.2). Lower seroprevalence was identified in schoolchildren who reported following the recommended protective measures against covid-19. CONCLUSION: Seroprevalence of SARS-CoV-2 antibodies is found mainly in the most socially vulnerable schoolchildren. This study can contribute to support public policies that reinforce the importance of suspending face-to-face classes and developing strategies aimed at protective measures and monitoring of the serological status of those who have not yet been included in the vaccination schedule.

Dissecting the KNDy hypothesis: KNDy neuron-derived kisspeptins are dispensable for puberty but essential for preserved female fertility and gonadotropin pulsatility.

BACKGROUND: Kiss1 neurons in the hypothalamic arcuate-nucleus (ARC) play key roles in the control of GnRH pulsatility and fertility. A fraction of ARC Kiss1 neurons, termed KNDy, co-express neurokinin B (NKB; encoded by Tac2). Yet, NKB- and Kiss1-only neurons are also found in the ARC, while a second major Kiss1-neuronal population is present in the rostral hypothalamus. The specific contribution of different Kiss1 neuron sub-sets and kisspeptins originating from them to the control of reproduction and eventually other bodily functions remains to be fully determined. METHODS: To tease apart the physiological roles of KNDy-born kisspeptins, conditional ablation of Kiss1 in Tac2-expressing cells was implemented in vivo. To this end, mice with Tac2 cell-specific Kiss1 KO (TaKKO) were generated and subjected to extensive reproductive and metabolic characterization. RESULTS: TaKKO mice displayed reduced ARC kisspeptin content and Kiss1 expression, with greater suppression in females, which was detectable at infantile-pubertal age. In contrast, Tac2/NKB levels were fully preserved. Despite the drop of ARC Kiss1/kisspeptin, pubertal timing was normal in TaKKO mice of both sexes. However, young-adult TaKKO females displayed disturbed LH pulsatility and sex steroid levels, with suppressed basal LH and pre-ovulatory LH surges, early-onset subfertility and premature ovarian insufficiency. Conversely, testicular histology and fertility were grossly conserved in TaKKO males. Ablation of Kiss1 in Tac2-cells led also to sex-dependent alterations in body composition, glucose homeostasis, especially in males, and locomotor activity, specifically in females. CONCLUSIONS: Our data document that KNDy-born kisspeptins are dispensable/compensable for puberty in both sexes, but required for maintenance of female gonadotropin pulsatility and fertility, as well as for adult metabolic homeostasis. SIGNIFICANCE STATEMENT: Neurons in the hypothalamic arcuate nucleus (ARC) co-expressing kisspeptins and NKB, named KNDy, have been recently suggested to play a key role in pulsatile secretion of gonadotropins, and hence reproduction. However, the relative contribution of this Kiss1 neuronal-subset, vs. ARC Kiss1-only and NKB-only neurons, as well as other Kiss1 neuronal populations, has not been assessed in physiological settings. We report here findings in a novel mouse-model with elimination of KNDy-born kisspeptins, without altering other kisspeptin compartments. Our data highlights the heterogeneity of ARC Kiss1 populations and document that, while dispensable/compensable for puberty, KNDy-born kisspeptins are required for proper gonadotropin pulsatility and fertility, specifically in females, and adult metabolic homeostasis. Characterization of this functional diversity is especially relevant, considering the potential of kisspeptin-based therapies for management of human reproductive disorders.

Seroprevalence of SARS-CoV-2 antibodies in schoolchildren in the city of São Paulo, 2020 / Soroprevalência de anticorpos do vírus SARS-CoV-2 em escolares no município de São Paulo, 2020

ABSTRACT OBJECTIVE To estimate seroprevalence of SARS-CoV-2 antibodies in schoolchildren aged 4 to 14 years living in the city of São Paulo, according to clinical, demographic, epidemiological, and social variables, during the school closure period as a measure against covid-19 spread. METHODS A serological survey was made in September 2020 with a random sample stratified by school system (municipal public, state public and private) type. A venous blood sample was collected using the Wondfo SARS-CoV-2 Antibody Test (lateral flow method) for detection of total SARS-CoV-2 virus antibodies. Semi-structured questionnaires were applied to collect clinical, demographic, social, and epidemiological data. RESULTS Seroprevalence of SARS-CoV-2 antibodies in schoolchildren was of 16.6% (95%CI 15.4-17.8). The study found higher seroprevalence in the municipal (18.5%; 95%CI 16.6-20.6) and state (16.2%; 95%CI 14.4-18.2) public school systems compared to the private school system (11.7; 95%CI 10.0-13.7), among black and brown students (18.4%; 95%CI 16.8-20.2) and in the most vulnerable social stratum (18.5 %;95%CI 16.9-20.2). Lower seroprevalence was identified in schoolchildren who reported following the recommended protective measures against covid-19. CONCLUSION Seroprevalence of SARS-CoV-2 antibodies is found mainly in the most socially vulnerable schoolchildren. This study can contribute to support public policies that reinforce the importance of suspending face-to-face classes and developing strategies aimed at protective measures and monitoring of the serological status of those who have not yet been included in the vaccination schedule.

RESUMO OBJETIVO Estimar a soroprevalência de anticorpos do vírus SARS-CoV-2 em escolares de quatro a 14 anos de idade residentes no município de São Paulo, segundo variáveis clínicas, demográficas, epidemiológicas e sociais, durante o período de fechamento das escolas como medida de controle da covid-19. MÉTODOS Realizou-se um inquérito sorológico em setembro de 2020 com amostra aleatória estratificada por tipo de rede de ensino (pública municipal, pública estadual e privada). Foi coletada amostra de sangue venoso utilizando-se o teste de imunoensaio de fluxo lateral da fabricante Wondfo para detecção de anticorpos totais contra o vírus SARS-CoV-2. Aplicaram-se questionários semiestruturados para o levantamento de dados clínicos, demográficos, sociais e epidemiológicos. RESULTADOS A soroprevalência de anticorpos do vírus SARS-CoV-2 em escolares foi de 16,6% (IC95% 15,4-17,8). O estudo encontrou soroprevalências mais elevadas na rede pública municipal (18,5%; IC95% 16,6-20,6) e estadual (16,2%; IC95% 14,4-18,2) em relação à rede privada (11,7; IC95% 10,0-13,7) e entre escolares da raça/cor preta e parda (18,4%; IC95% 16,8-20,2) e no estrato social mais vulnerável (18,5%; IC95% 16,9-20,2). A pesquisa identificou menores soroprevalências nos escolares que relataram seguir as medidas recomendadas de proteção contra a covid-19. CONCLUSÃO A soroprevalência de anticorpos contra o vírus SARS-CoV-2 atinge principalmente os escolares socialmente mais vulneráveis. Este estudo pode contribuir para embasar políticas públicas que reforcem a importância da suspensão das aulas presenciais e da necessidade de estratégias de medidas de proteção e acompanhamento do status sorológico daqueles que ainda não foram contemplados no calendário vacinal.

Cannabidiol markedly alleviates skin and liver fibrosis.

Cannabidiol (CBD) has been suggested as a potential therapy for inflammatory and fibrotic diseases. Cannabidiol was demonstrated to reduce alcohol-induced liver inflammation and steatosis but its specific activity on the fibrotic process was not investigated. Herein, the antifibrotic effects of cannabidiol in the skin were analysed in vitro using NIH-3T3 fibroblasts and human dermal fibroblasts and in vivo using the bleomycin-induced model of skin fibrosis. In a second model, non-alcoholic liver fibrosis was induced in mice by CCl4 exposure. Cannabidiol was administered daily, intraperitoneally in mice challenged with bleomycin and orally in CCl4 mice, and skin and liver fibrosis and inflammation were assessed by immunochemistry. Cannabidiol inhibited collagen gene transcription and synthesis and prevented TGFß-and IL-4 induced fibroblast migration. In the bleomycin model, cannabidiol prevented skin fibrosis and collagen accumulation around skin blood vessels, and in the CCl4 model cannabidiol significantly attenuated liver fibrosis measured by picrosirius red and Tenascin C staining and reduced T cell and macrophage infiltration. Altogether, our data further support the rationale of the medicinal use of this cannabinoid, as well as cannabis preparations containing it, in the management of fibrotic diseases including Systemic Sclerosis and Non-Alcoholic Fatty Liver Disease.

Dicer ablation in Kiss1 neurons impairs puberty and fertility preferentially in female mice.

Kiss1 neurons, producing kisspeptins, are essential for puberty and fertility, but their molecular regulatory mechanisms remain unfolded. Here, we report that congenital ablation of the microRNA-synthesizing enzyme, Dicer, in Kiss1 cells, causes late-onset hypogonadotropic hypogonadism in both sexes, but is compatible with pubertal initiation and preserved Kiss1 neuronal populations at the infantile/juvenile period. Yet, failure to complete puberty and attain fertility is observed only in females. Kiss1-specific ablation of Dicer evokes disparate changes of Kiss1-cell numbers and Kiss1/kisspeptin expression between hypothalamic subpopulations during the pubertal-transition, with a predominant decline in arcuate-nucleus Kiss1 levels, linked to enhanced expression of its repressors, Mkrn3, Cbx7 and Eap1. Our data unveil that miRNA-biosynthesis in Kiss1 neurons is essential for pubertal completion and fertility, especially in females, but dispensable for initial reproductive maturation and neuronal survival in both sexes. Our results disclose a predominant miRNA-mediated inhibitory program of repressive signals that is key for precise regulation of Kiss1 expression and, thereby, reproductive function.

Kappa-Opioid Receptor Blockade Ameliorates Obesity Caused by Estrogen Withdrawal via Promotion of Energy Expenditure through mTOR Pathway.

Weight gain is a hallmark of decreased estradiol (E2) levels because of menopause or following surgical ovariectomy (OVX) at younger ages. Of note, this weight gain tends to be around the abdomen, which is frequently associated with impaired metabolic homeostasis and greater cardiovascular risk in both rodents and humans. However, the molecular underpinnings and the neuronal basis for these effects remain to be elucidated. The aim of this study is to elucidate whether the kappa-opioid receptor (k-OR) system is involved in mediating body weight changes associated with E2 withdrawal. Here, we document that body weight gain induced by OVX occurs, at least partially, in a k-OR dependent manner, by modulation of energy expenditure independently of food intake as assessed in Oprk1-/-global KO mice. These effects were also observed following central pharmacological blockade of the k-OR system using the k-OR-selective antagonist PF-04455242 in wild type mice, in which we also observed a decrease in OVX-induced weight gain associated with increased UCP1 positive immunostaining in brown adipose tissue (BAT) and browning of white adipose tissue (WAT). Remarkably, the hypothalamic mTOR pathway plays an important role in regulating weight gain and adiposity in OVX mice. These findings will help to define new therapies to manage metabolic disorders associated with low/null E2 levels based on the modulation of central k-OR signaling.

Connecting nutritional deprivation and pubertal inhibition via GRK2-mediated repression of kisspeptin actions in GnRH neurons.

BACKGROUND: Perturbations in the timing of puberty, with potential adverse consequences in later health, are increasingly common. The underlying neurohormonal mechanisms are unfolded, but nutritional alterations are key contributors. Efforts to unveil the basis of normal puberty and its metabolic control have focused on mechanisms controlling expression of Kiss1, the gene encoding the puberty-activating neuropeptide, kisspeptin. However, other regulatory phenomena remain ill-defined. Here, we address the putative role of the G protein-coupled-receptor kinase-2, GRK2, in GnRH neurons, as modulator of pubertal timing via repression of the actions of kisspeptin, in normal maturation and conditions of nutritional deficiency. METHODS: Hypothalamic RNA and protein expression analyses were conducted in maturing female rats. Pharmacological studies involved central administration of GRK2 inhibitor, ßARK1-I, and assessment of gonadotropin responses to kisspeptin or phenotypic and hormonal markers of puberty, under normal nutrition or early subnutrition in female rats. In addition, a mouse line with selective ablation of GRK2 in GnRH neurons, aka G-GRKO, was generated, in which hormonal responses to kisspeptin and puberty onset were monitored, in normal conditions and after nutritional deprivation. RESULTS: Hypothalamic GRK2 expression increased along postnatal maturation in female rats, especially in the preoptic area, where most GnRH neurons reside, but decreased during the juvenile-to-pubertal transition. Blockade of GRK2 activity enhanced Ca+2 responses to kisspeptin in vitro, while central inhibition of GRK2 in vivo augmented gonadotropin responses to kisspeptin and advanced puberty onset. Postnatal undernutrition increased hypothalamic GRK2 expression and delayed puberty onset, the latter being partially reversed by central GRK2 inhibition. Conditional ablation of GRK2 in GnRH neurons enhanced gonadotropin responses to kisspeptin, accelerated puberty onset, and increased LH pulse frequency, while partially prevented the negative impact of subnutrition on pubertal timing and LH pulsatility in mice. CONCLUSIONS: Our data disclose a novel pathway whereby GRK2 negatively regulates kisspeptin actions in GnRH neurons, as major regulatory mechanism for tuning pubertal timing in nutritionally-compromised conditions.

Preclinical Investigation in Neuroprotective Effects of the GPR55 Ligand VCE-006.1 in Experimental Models of Parkinson's Disease and Amyotrophic Lateral Sclerosis.

Cannabinoids act as pleiotropic compounds exerting, among others, a broad-spectrum of neuroprotective effects. These effects have been investigated in the last years in different preclinical models of neurodegeneration, with the cannabinoid type-1 (CB1) and type-2 (CB2) receptors concentrating an important part of this research. However, the issue has also been extended to additional targets that are also active for cannabinoids, such as the orphan G-protein receptor 55 (GPR55). In the present study, we investigated the neuroprotective potential of VCE-006.1, a chromenopyrazole derivative with biased orthosteric and positive allosteric modulator activity at GPR55, in murine models of two neurodegenerative diseases. First, we proved that VCE-006.1 alone could induce ERK1/2 activation and calcium mobilization, as well as increase cAMP response but only in the presence of lysophosphatidyl inositol. Next, we investigated this compound administered chronically in two neurotoxin-based models of Parkinson's disease (PD), as well as in some cell-based models. VCE-006.1 was active in reversing the motor defects caused by 6-hydroxydopamine (6-OHDA) in the pole and the cylinder rearing tests, as well as the losses in tyrosine hydroxylase-containing neurons and the elevated glial reactivity detected in the substantia nigra. Similar cytoprotective effects were found in vitro in SH-SY5Y cells exposed to 6-OHDA. We also investigated VCE-006.1 in LPS-lesioned mice with similar beneficial effects, except against glial reactivity and associated inflammatory events, which remained unaltered, a fact confirmed in BV2 cells treated with LPS and VCE-006.1. We also analyzed GPR55 in these in vivo models with no changes in its gene expression, although GPR55 was down-regulated in BV2 cells treated with LPS, which may explain the lack of efficacy of VCE-006.1 in such an assay. Furthermore, we investigated VCE-006.1 in two genetic models of amyotrophic lateral sclerosis (ALS), mutant SOD1, or TDP-43 transgenic mice. Neither the neurological decline nor the deteriorated rotarod performance were prevented with this compound, and the same happened with the elevated microglial and astroglial reactivities, albeit modest spinal motor neuron preservation was achieved in both models. We also analyzed GPR55 in these in vivo models and found no changes in both TDP-43 transgenic and mSOD1 mice. Therefore, our findings support the view that targeting the GPR55 may afford neuroprotection in experimental PD, but not in ALS, thus stressing the specificities for the development of cannabinoid-based therapies in the different neurodegenerative disorders.

Small extracellular vesicle-mediated targeting of hypothalamic AMPKα1 corrects obesity through BAT activation.

Current pharmacological therapies for treating obesity are of limited efficacy. Genetic ablation or loss of function of AMP-activated protein kinase alpha 1 (AMPKα1) in steroidogenic factor 1 (SF1) neurons of the ventromedial nucleus of the hypothalamus (VMH) induces feeding-independent resistance to obesity due to sympathetic activation of brown adipose tissue (BAT) thermogenesis. Here, we show that body weight of obese mice can be reduced by intravenous injection of small extracellular vesicles (sEVs) delivering a plasmid encoding an AMPKα1 dominant negative mutant (AMPKα1-DN) targeted to VMH-SF1 neurons. The beneficial effect of SF1-AMPKα1-DN-loaded sEVs is feeding-independent and involves sympathetic nerve activation and increased UCP1-dependent thermogenesis in BAT. Our results underscore the potential of sEVs to specifically target AMPK in hypothalamic neurons and introduce a broader strategy to manipulate body weight and reduce obesity.

Prevalence evolution of SARS-CoV-2 infection in the city of São Paulo, 2020-2021.

OBJECTIVE: To estimate the evolution of the prevalence of SARS-CoV-2 virus infection among residents aged 18 years or over in the municipality of São Paulo. METHODS: This is a population-based household survey conducted every 15 days, between June and September 2020, and January and February 2021. In total, the study comprised 11 phases. The presence of antibodies against SARS-CoV-2 was identified in venous blood using a lateral flow test, Wondfo Biotech. In the last phase, the researchers combined it with an immunoenzymatic test, Euroimmun. The participants also answered a semi-structured questionnaire on sociodemographic and economic factors, and on social distancing measures. Prevalence estimates and the 95% confidence interval were estimated according to regions, Human Development Index, sex, age group, ethnicity, education, income, and variables associated with risk or prevention of infection. To compare the frequencies among the categories of each variable, the chi-square test with Rao-Scott correction was used, considering a significance level of 5%. RESULTS: In total, 23,397 individuals were interviewed and had their samples collected. The estimated prevalence of antibodies against SARS-CoV-2 ranged from 9.7% (95%CI: 7.9-11.8%) to 25.0% (95%CI: 21.7-28.7). The prevalence of individuals with antibodies against the virus was higher among black and brown people, people with lower schooling and income, and among residents of regions with lower Human Development Index. The lowest prevalences were associated with recommended measures of disease protection. The proportion of asymptomatic infection was 45.1%. CONCLUSION: The estimated prevalence of the infection was lower than the cumulative incidence variation, except for the last phase of the study. The differences in prevalence estimates observed among subpopulations showed social inequality as a risk of infection. The lower prevalence observed among those who could follow prevention measures reinforce the need to maintain social distancing measures as a way to prevent SARS-CoV-2 infection.

Effects of substituting sweet potato flour for ground corn on performance, feeding behavior, and metabolism of dairy cows.

The objective of this study was to evaluate the effect of substituting sweet potato flour for ground corn in rations fed to lactating dairy on milk yield and composition, blood metabolites, and feeding behavior. Twenty lactating Holstein cows from 30 to 60 days postpartum were randomly assigned to one of two groups (n = 10 each) and used in a cross-over design trial with two treatments: a standard concentrate with ground corn as an energy source or experimental concentrate with sweet potato flour (SPF) replacing all of the ground corn. Each of the 35-day periods consisted of 14 days for adaptation to diet and 21 days for data and sample collection. Milk yield, dry matter intake (DMI), and feeding behavior were evaluated daily throughout the trial. Milk samples were collected weekly and blood samples were collected every 3 days. Milk was analyzed for fat, protein, lactose, and total solid constituents. Blood was analyzed for glucose, non-esterified fatty acid (NEFA), gamma-glutamyl transferase (GGT), aspartate aminotransferase (AST), total protein (TP), albumin, and urea concentrations. Milk yield (P = 0.62) and composition (fat: P = 0.71; protein: P = 0.12; lactose: P = 0.82; total solids: P = 0.56) were not affected by dietary treatments. There were no differences between treatments in DMI or meal frequency, but total eating time (P = 0.001), feeding time (P = 0.001), and meal duration (P = 0.001) was higher for control compared with SPF. However, feeding rate (P = 0.001) and serum urea concentration (P = 0.001) were higher for SPF compared with control. No differences were observed in serum metabolites and enzymes measured among treatments. The results of this trial indicate that SPF can be substituted for ground corn without impairing the performance, feeding behavior, and metabolism in dairy cows.

In1-Ghrelin Splicing Variant as a Key Element in the Pathophysiological Association Between Obesity and Prostate Cancer.

CONTEXT: Recent studies emphasize the importance of considering the metabolic status to develop personalized medicine approaches. This is especially relevant in prostate cancer (PCa), wherein the diagnostic capability of prostate-specific antigen (PSA) dramatically drops when considering patients with PSA levels ranging from 3 to 10 ng/mL, the so-called grey zone. Hence, additional noninvasive diagnostic and/or prognostic PCa biomarkers are urgently needed, especially in the metabolic-status context. OBJECTIVE: To assess the potential relation of urine In1-ghrelin (a ghrelin-splicing variant) levels with metabolic-related/pathological conditions (eg, obesity, diabetes, body mass index, insulin and glucose levels) and to define its potential clinical value in PCa (diagnostic/prognostic capacity) and relationship with PCa risk in patients with PSA in the grey zone. METHODS: Urine In1-ghrelin levels were measured by radioimmunoassay in a clinically, metabolically, pathologically well-characterized cohort of patients without (n = 397) and with (n = 213) PCa with PSA in the grey zone. RESULTS: Key obesity-related factors associated with PCa risk (BMI, diabetes, glucose and insulin levels) were strongly correlated to In1-ghrelin levels. Importantly, In1-ghrelin levels were higher in PCa patients compared to control patients with suspect of PCa but negative biopsy). Moreover, high In1-ghrelin levels were associated with increased PCa risk and linked to PCa aggressiveness (eg, tumor stage, lymphovascular invasion). In1-ghrelin levels added significant diagnostic value to a clinical model consisting of age, suspicious digital rectal exam, previous biopsy, and PSA levels. Furthermore, a multivariate model consisting of clinical and metabolic variables, including In1-ghrelin levels, showed high specificity and sensitivity to diagnose PCa (area under the receiver operating characteristic curve = 0.740). CONCLUSIONS: Urine In1-ghrelin levels are associated with obesity-related factors and PCa risk and aggressiveness and could represent a novel and valuable noninvasive PCa biomarker, as well as a potential link in the pathophysiological relationship between obesity and PCa.

Zootechnical and health performance of Holstein x Gir crossbred calves.

Crossbreeding is used to increase production and disease resistance in adult animals, and there is no research to assess the performance of animals in the early stages. The aim of this study was to evaluate the zootechnical and health performance of Holstein x Gir calves (½ HG: ½ Dutch ½ Gir and ¾ HG: ¾ Dutch » Gir), from birth to 80 days of age, and compare metabolic parameters between groups. In this sense, calves were monitored for zootechnical parameters; epidemiological indexes such as morbidity, mortality, recurrence of diarrhea, pneumonia, and other diseases; as well as serum concentrations of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, gamma-glutamyl transferase, total proteins, cholesterol, triglycerides, albumin, urea, and paraoxonase1 (PON1). ¾ HG calves showed higher morbidity for diarrhea and remained with diarrhea for longer compared to ½ HG calves, and this was reflected in the average daily weight gain until the 42nd day, with ½ HG calves performing better. There were no differences regarding passive immune transfer between groups, as well as no differences in morbidity and mortality from pneumonia. Regarding biochemical analyses, a difference was found only in the concentrations of PON1, which were higher in ¾ HG calves. The findings show that blood degree influences the occurrence and duration of diarrhea, negatively impacting the zootechnical performance of the animals. Crossbreeding bulls with zebu cattle can be an alternative to increase calf resistance and reduce diarrhea, thus lowering economic losses and improving animal performance.

Δ9-Tetrahydrocannabinolic Acid markedly alleviates liver fibrosis and inflammation in mice.

BACKGROUND: Non-alcoholic fatty liver disease (NAFLD) is the most common liver disease in the Western world, and it is closely associated to obesity, type 2 diabetes mellitus, and dyslipidemia. Medicinal cannabis and some neutral cannabinoids have been suggested as a potential therapy for liver diseases. HYPOTHESIS: Δ9-tetrahydrocannabinolic acid (Δ9-THCA), the non-psychotropic precursor of Δ9-THC, is one of the most abundant cannabinoids presents in Cannabis Sativa. However, its biological activities have been poorly investigated. Herein, we studied the antifibrotic and antiinflammatory activities of Δ9-THCA in two different animal models of liver injury, providing a rationale for additional studies on the medicinal use of this cannabinoid in the treatment of liver fibrosis and the management of NAFLD. STUDY DESIGN: The antifibrotic activity of Δ9-THCA in vitro was investigated in the cell lines LX-2 and NIH-3T3-Col1A2-luc. Non-alcoholic liver fibrosis was induced in mice by CCl4 treatment or, alternatively, by 23-week high fat diet (HFD) feeding. Δ9-THCA was administered daily intraperitoneally during the CCl4 treatment or during the last 3 weeks in HFD-fed mice. METHODS: TGFß-induced profibrotic gene expression was analyzed by luciferase and qPCR assays. Liver fibrosis and inflammation were assessed by immunochemistry and qPCR. Blood glucose, insulin, leptin and triglyceride levels were measured in HFD mice. RESULTS: Δ9-THCA inhibited the expression of Tenascin C (TNC) and Col3A1 induced by TGFß in LX-2 cells and the transcriptional activity of the Col1A2 promoter in fibroblasts. Δ9-THCA significantly attenuated CCl4-induced liver fibrosis and inflammation and reduced T cell and macrophage infiltration. Mice fed HFD for 23 weeks developed severe obesity (DIO), fatty liver and marked liver fibrosis, accompanied by immune cell infiltration. Δ9-THCA, significantly reduced body weight and adiposity, improved glucose tolerance, and drastically attenuated DIO-induced liver fibrosis and immune cell infiltration. CONCLUSIONS: Δ9-THCA prevents TGFß-induced fibrotic markers in vitro and liver inflammation and fibrogenesis in vivo, providing a rationale for additional studies on the medicinal use of this cannabinoid, as well as cannabis preparations containing it, for the treatment of liver fibrosis and the management of NAFLD.

Prevalence evolution of SARS-CoV-2 infection in the city of São Paulo, 2020-2021

ABSTRACT OBJECTIVE To estimate the evolution of the prevalence of SARS-CoV-2 virus infection among residents aged 18 years or over in the municipality of São Paulo. METHODS This is a population-based household survey conducted every 15 days, between June and September 2020, and January and February 2021. In total, the study comprised 11 phases. The presence of antibodies against SARS-CoV-2 was identified in venous blood using a lateral flow test, Wondfo Biotech. In the last phase, the researchers combined it with an immunoenzymatic test, Euroimmun. The participants also answered a semi-structured questionnaire on sociodemographic and economic factors, and on social distancing measures. Prevalence estimates and the 95% confidence interval were estimated according to regions, Human Development Index, sex, age group, ethnicity, education, income, and variables associated with risk or prevention of infection. To compare the frequencies among the categories of each variable, the chi-square test with Rao-Scott correction was used, considering a significance level of 5%. RESULTS In total, 23,397 individuals were interviewed and had their samples collected. The estimated prevalence of antibodies against SARS-CoV-2 ranged from 9.7% (95%CI: 7.9-11.8%) to 25.0% (95%CI: 21.7-28.7). The prevalence of individuals with antibodies against the virus was higher among black and brown people, people with lower schooling and income, and among residents of regions with lower Human Development Index. The lowest prevalences were associated with recommended measures of disease protection. The proportion of asymptomatic infection was 45.1%. CONCLUSION The estimated prevalence of the infection was lower than the cumulative incidence variation, except for the last phase of the study. The differences in prevalence estimates observed among subpopulations showed social inequality as a risk of infection. The lower prevalence observed among those who could follow prevention measures reinforce the need to maintain social distancing measures as a way to prevent SARS-CoV-2 infection.

Use of sulfonamides for the treatment of bovine neonatal diarrhea: clinical and performance parameters / Utilização de sulfonamidas para o tratamento de diarreia neonatal bovina: parâmetros clínicos e zootécnicos

This study was developed to evaluate the clinical, hematological, and performance parameters of calves affected by diarrhea caused by the bacterial enteric pathogen Escherichia coli, treated with three different protocols containing sulfonamides. Fourteen Holstein calves were monitored from birth to 60 days of age, and divided into 3 groups. Group 1 (n=5) animals were treated with 80 g of Kaopek®, Ibasa, Brazil, administered orally (BM), dissolved in 160 ml of water every 24 h; Group 2 (n=4) received IM sulfadiazine and trimethoprim (Ibatrim®, Ibasa, Brazil), and Group 3 (n=5) animals were treated with 80 g of Kaopek®, Ibasa, Brazil, which is made up of 16 g of phthalylsulfathiazole, associated with 2.28 g neomycin sulfate, 1.6 g of pectin, and 80 g of kaolin, dissolved in 160 ml of water and administered every 24 h by mouth, in addition to sulfadiazine and trimethoprim (Ibatrim®, Ibasa, Brazil), at a daily dose of 16 mg/kg live weight, through IM injection. During the study, the animals were clinically evaluated, and once they were diagnosed with diarrhea, feces samples were collected to identify the bacterial enteric pathogen, antibiogram, polymerase chain reaction (PCR), and coproparasitological exams. Blood samples were collected to evaluate the hematological profile, and the performance profile was monitored weekly. In the clinical examination, all calves presented a reduction in body temperature (<39.2°C) and some improvement in hydration after treatment (p=0,31). However, group 2 had a better concentration of lymphocytes and TP concerning the other animals, as well as better performance. Besides, E.coli was detected in 100% of feces samples. Thus, the therapeutic protocols with sulfonamides used to treat bovine neonatal diarrhea were effective in the clinical improvement of the animals. Nonetheless, the protocol with systemic therapy using sulfadiazine and trimethoprim (Ibatrim®, Ibasa, Brazil) intramuscularly, provided better performance, with better weight gain, and body development of the animals.(AU)

Este estudo foi desenvolvido com o objetivo de avaliar parâmetros clínicos, hematológicos e zootécnicos de bezerras acometidas por diarreia provocada pelo agente bacteriano Escherichia coli, tratadas com três diferentes protocolos contendo sulfonamidas. Quatorze bezerras da raça Holandês foram monitoradas do nascimento até os 60 dias de vida e divididas em 3 grupos: Grupo 1 (n=5), animais tratados com a dose de 80g de Kaopek®, Ibasa, Brasil, por via oral (VO), dissolvido em 160ml de água a cada 24 horas; Grupo 2 (n=4) receberam sulfadiazina e trimetropim (Ibatrim®, Ibasa, Brasil), na dose de 16mg/Kg de peso vivo, por dia, por via intramuscular (IM); Grupo 3 (n=5) 80g de Kaopek®, Ibasa, Brasil, por VO dissolvido em 160ml de água, a cada 24 horas e com 16mg/Kg de peso vivo, por dia, de sulfadiazina e trimetoprim (Ibatrim®, Ibasa, Brasil), por via IM. Durante o estudo, os animais foram avaliados clinicamente e a partir do diagnóstico de diarreia foram coletadas amostras de fezes para a identificação do agente bacteriano, antibiograma, reação em cadeia da polimerase (PCR) e exames coproparasitológicos. Amostras de sangue foram coletadas para avaliação do perfil hematológico e o perfil zootécnico foi acompanhado semanalmente. No exame clínico, todas as bezerras apresentaram redução na temperatura corporal (<39,2°C) e melhora na hidratação após o período de tratamento (p=0,31), porém o grupo 2 obteve maior concentração de linfócitos e PPT em relação aos demais, assim como melhor desempenho zootécnico, além disso em 100% das amostras de fezes foi detectado E. coli. Sendo assim, os protocolos terapêuticos com sulfonamidas utilizados para o tratamento da diarreia neonatal bovina foram eficazes na melhora clínica dos animais. Porém, o protocolo com terapia sistêmica com sulfadiazina e trimetoprim (Ibatrim®, Ibasa, Brasil) por via intramuscular proporcionou um melhor desempenho zootécnico, com melhor ganho de peso e desenvolvimento corporal dos animais.(AU)