Assessment of SARS-CoV-2 infection in a basic health service unit in Araçatuba, São Paulo, Brazil / Incidência de infecção por SARS-Cov-2 em uma unidade básica de Saúde de Araçatuba, São Paulo, Brasil

Objective: to describe risk factors associated with SARS-CoV-2 infection. Methods: this is a retrospective descriptive cross-sectional study aimed at describing the epidemiological profile of laboratory and clinical diagnosis of unvaccinated patients seen at a basic health unit in Araçatuba ­ SP, infected with SARS-CoV-2 between June 2020 and January 2021.The results were analyzed through inferential and descriptive statistics. Additionally, Chi-square and Fisher exact tests were used (p<0.05). Results: of 313 patients, 128 were positive for COVID-19, with 68.75% diagnosed by RT-PCR and the others by immunochromatography. Women were 51.56% of those infected with adults corresponding to the main age group (76.56%), and 57% of patients had only a basic educational level concluded. A total of 88.26% of the patients progressed to cure without complications; eight patients died, most of whom were men and elderly. Of the variables analyzed for positive/negative outcomes, only "basic educational level" was significant for a positive result(p=0.0019). Conclusion: the deaths of infected patients may be associated with the existence of at least one comorbidity and advanced age of men.

Objetivo: descrever os fatores de risco associados com a infecção por SARS-CoV-2. Métodos: trata-se de um estudo descritivo retrospectivo e transversal, voltado a descrever o perfil epidemiológico de diagnóstico laboratorial e clínico, de pacientes não vacinados, atendidos em uma unidade básica de saúde de Araçatuba-SP, infectados por SARS-CoV-2, no período entre junho de 2020 e janeiro de 2021. Os resultados foram analisados por estatística inferencial e descritiva. Adicionalmente, foram aplicados os testes de Qui-quadrado e exato de Fisher (p<0.05). Resultados: dos 313 pacientes, 128 apresentaram resultado positivo para COVID-19, com 68,75% diagnosticados por RT-PCR e o restante por imunocromatografia. Mulheres foram 51,56% dos infectados, com adultos correspondendo à principal faixa etária (76,56%), 57% dos pacientes apresentavam apenas o nível educacional básico concluído. O quadro de 88.26%dos pacientes evoluiu para cura sem complicações;8 pacientes foram a óbito, sendo, em sua maioria, homens e idosos. Das variáveis analisadas para grau de dependência de resultado positivo/negativo, apenas "nível escolar básico" apresentou resultado significante para resultado positivo (p=0.0019). Conclusão: os óbitos dos pacientes infectados podem ser associados à existência de, pelo menos, uma comorbidade e à idade avançada de homens.

Cytotoxicity of Structurally Diverse Anthranoids and Correlation with Mechanism of Action and Side Effects.

The purpose of this contribution is to evaluate the cytotoxicity and apoptosis inducing ability of structurally diverse anthraquinones to establish a relationship between structure and toxicity. Besides the wide spread use of anthraquinones in pharmacological drugs for constipation and non-prescription dietary supplements for weight loss, extracts are still commercialized as crude extracts and long-term side effects are still relevant. In this work we developed a method to quantify the cascarosides isolated from Rhamnus purshiana (Cascara Sagrada) using LC-MS/MS and evaluated the effects of this extract and isolated compounds on cellular viability using NOK-SI, HeLa, and T98G cell lineages. Apoptosis inducing ability was also analyzed via evaluating key-proteins involved in apoptosis pathways. Using cascarosides isolated from bark extracts, we found that the presence of glucose moieties in the chemical structure reduced the toxicity. This communication reviewed the mechanisms of action, toxicity of anthraquinones and correlated the toxicity with chemical structures of cascarosides. Results indicate that cascarosides-enriched cascara extract, as well as glycosylated anthraquinones, may have some beneficial effects for laxative action of herbal medicines. Considering our results, a cascarosides-enrichment in cascara extract is recommended.

Oxytocin effects on osteoblastic differentiation of Bone Marrow Mesenchymal Stem Cells from adult and aging female Wistar rats.

Recently, it has been suggested that oxytocin (OT) might play a role in the control of bone remodeling and in bone health of young and adult females. The purpose of this study was to evaluate the effect of osteogenic medium (OM) plus OT (OM + OT; 100 nmol/L) on osteoblastic differentiation of bone marrow mesenchymal stem cells (BMMSCs) from cyclic adult (12 months old) and acyclic aging (24 months old) female Wistar rats. After 14 days, OM + OT increased the oxytocin and oxytocin receptor in the BMMSCs from animals of both age groups relative to OM controls. Alkaline phosphatase activity was higher in the OM + OT than OM group in BMMSCs from 24-month-old female rats. OM + OT improved osteogenic differentiation, observed by anticipated mineralization and increased gene expression of bone morphogenetic protein 2, bone sialoprotein, osteopontin and osteocalcin in both aged relative to OM controls. These findings suggest a role for OT as an adjuvant to induce osteoblastic differentiation of BMMSCs from aged female rat.

Effects of strength training on osteogenic differentiation and bone strength in aging female Wistar rats.

The effects of strength training (ST) on the mechanical bone strength and osteogenic differentiation of bone marrow mesenchymal stromal cells (BMSCs) from adult, aged and exercised aged rats were determined. The exercised aged animals displayed higher values of areal bone mineral density, compression test, alkaline phosphatase activity (ALP) and biological mineralization, while oil red O staining for adipocytes was lower. ST increased gene expression of runt-related transcription factor 2 (Runx2), osterix (Osx) as well as bone matrix protein expression, and reduced expression of peroxisome proliferator-activated receptor gamma (Pparγ). The production of pro-inflammatory cytokine tumor necrosis factor alpha (TNF-α) was lower in BMSCs of the aged exercised group. The ST practice was able to improve the bone mechanical properties in aged female rats, increasing the potential for osteogenic differentiation of BMSCs, reducing the adipogenic differentiation and pro-inflammatory cytokine level. In summary, the data achieved in this study showed that strength training triggers physiological responses that result in changes in the bone microenvironment and bring benefits to biomechanical parameters of bone tissue, which could reduce the risk of fractures during senescent.

The effects of strength training and raloxifene on bone health in aging ovariectomized rats.

The aim of this study was to investigate the effects of strength training (ST) and raloxifene (Ral), alone or in combination, on the prevention of bone loss in an aging estrogen-deficient rat model. Aging Wistar female rats were ovariectomized at 14months and allocated to four groups: (1) non-trained and treated with vehicle, NT-Veh; (2) strength training and treated with vehicle, ST-Veh; (3) non-trained and treated with raloxifene, NT-Ral; and (4) strength training and treated with raloxifene, ST-Ral. ST was performed on a ladder three times per week and Ral was administered daily by gavage (1mg/kg/day), both for 120days. Areal bone mineral density (aBMD), strength, microarchitecture, and biomarkers (osteocalcin, OCN; osteoprotegerin, OPG; and tartrate-resistant acid phosphatase, TRAP) were assessed. Immunohistochemistry was performed for runt-related transcription factor 2 (RUNX2), osterix (OSX), OCN, OPG, TRAP, and receptor activator of nuclear factor kappa-B ligand (RANKL). The rats that performed ST (ST-Veh) or were treated with Ral (NT-Ral) showed significant improvements in aBMD (p=0.001 and 0.004), bone strength (p=0.001), and bone microarchitecture, such as BV/TV (%) (p=0.001), BS/TV (mm(2)/mm(3)) (p=0.023 and 0.002), Conn.Dn (1/mm(3)) (p=0.001), Tb.N (1/mm) (p=0.012 and 0.011), Tb.Th (1/mm) (p=0.001), SMI (p=0.001 and 0.002), Tb.Sp (p=0.001), and DA (p=0.002 and 0.007); there was also a significant decrease in plasma levels of OCN (p=0.001 and 0.002) and OPG (p=0.003 and 0.014), compared with animals in the NT-Veh group. Ral, with or without ST, promoted an increased immunolabeling pattern for RUNX2 (p=0.0105 and p=0.0006) and OSX (p=0.0105), but a reduced immunolabeling pattern for TRAP (p=0.0056) and RANKL (p=0.033 and 0.004). ST increased the immunolabeling pattern for RUNX2 (p=0.0105), and association with Ral resulted in an increased immunolabeling pattern for OPG (p=0.0034) and OCN (p=0.0024). In summary, ST and Ral administration in aged, estrogen-deficient Wistar female rats is associated with a decrease in bone turnover marker plasma levels, increased activity of cells that promote osteoblastogenesis, and decreased activity of cells that promote osteoclastogenesis; these are correlated with higher aBMD, bone strength, and bone microarchitecture at the femoral neck. The results indicate that strength training and Ral are potential tools to reduce the risk of fractures at clinically relevant sites.

Atuação da ocitocina no processo de diferenciação osteoblástica de ratas senis / Action of oxytocin in the process of osteoblast differentiation in senescent rats

Ocitocina (OT) é hormônio peptídico sintetizado no hipotálamo e armazenado na neurohipófise. A literatura evidencia atuação anabólica da OT no esqueleto de animais jovens. Em nosso laboratório estamos realizando pesquisas para estudar a ação da OT no metabolismo ósseo, durante o envelhecimento. Analisamos inicialmente o processo de remodelação óssea alveolar em ratas com 24 meses e detectamos ação importante da OT na maior formação óssea nesses animais, o que não foi observado em animais com 12 meses. Esses resultados evidenciaram a necessidade de investigação da ação da OT, visando à compreensão dos mecanismos envolvidos na maior formação óssea em organismos senis, no período caracterizado pela aciclicidade estral (estropausa – 24 meses) comparado com organismos adultos com ciclo estral regular (12 meses). Nesse estudo foi avaliada a influência da idade (12 e 24 meses) e a ação da OT associada a indutores osteogênicos clássicos durante a diferenciação em linhagem osteogênica de células-tronco mesenquimais (CTMs) a partir do estroma da medula óssea de fêmures de ratas Wistar, com 12 e 24 meses. Quatro grupos de cada idade foram formados, sendo: MC (meio de crescimento); MO (meio de crescimento suplementado com indutores osteogênicos); MCO (meio de crescimento + OT); MOO (meio de crescimento suplementado com indutores osteogênicos + OT). O ensaio com vermelho de alizarina demonstrou mineralização biológica a partir do 17º dia no grupo com 12 meses em ambos os grupos osteogênicos (MO e MOO), porém a adição de OT (MOO) promoveu a mineralização a partir do 14º dia. No grupo de 24 meses não foi detectada mineralização em nenhum dia experimental no grupo MO, porém a adição de OT no meio (MOO) promoveu a mineralização a partir do 17º dia. As escalas de expressão gênica de osterix, ocitocina e receptor de ocitocina foram menores no grupo 24 meses que no grupo 12 meses, assim como as proteínas de matriz analisadas (sialoproteína óssea, osteopontina e osteocalcina)...

Peripheral oxytocin (OT) has been shown to have direct effects on the skeleton, and to provide stimulus for the activity of osteoblasts, as an anabolic factor affecting bone mass. The effects of OT can vary with age. Here we evaluated the effects of OT on the differentiation of bone marrow mesenchymal stem cells, in 12- and 24-months-old Wistar female rats. Four treatment groups were formed for each age group: control (C) (growth medium); osteogenic medium (OM); control + oxytocin (C+OT) (growth medium + oxytocin) and osteogenic medium + oxytocin (OM+OT). Alizarin Red assay was performed and in cells from 12-month-old rats mineralization was detected on day 17 in OM. However, the addition of OT to osteogenic medium (OM+OT) provided biological mineralization on day 14. In cells from 24-month-old acyclic rats, the OM subgroup showed no mineralization; in the OM+OT subgroup mineralization was detected on day 17. Osterix, matrix protein (bone sialoprotein, osteopontin and osteocalcin) and oxytocin gene expression was lower in the 24-month-old than the 12-month-old group; these proteins were increased by OT. The oxytocin receptor was up-regulated only in the 12-month-old group and PGE2 showed an increase in the OM group than in the OM + OT group in both age groups. These findings suggest that OT shows different, differentiation-dependent actions, and is important in the biological mineralization performed by osteoblasts, mainly in cells of bone marrow mesenchymal stem cells from acyclic 24-month-old rats highlighting oxytocin as a potential therapy for control of osteoporosis...

Atuação da ocitocina no processo de diferenciação osteoblástica de ratas senis / Action of oxytocin in the process of osteoblast differentiation in senescent rats

Ocitocina (OT) é hormônio peptídico sintetizado no hipotálamo e armazenado na neurohipófise. A literatura evidencia atuação anabólica da OT no esqueleto de animais jovens. Em nosso laboratório estamos realizando pesquisas para estudar a ação da OT no metabolismo ósseo, durante o envelhecimento. Analisamos inicialmente o processo de remodelação óssea alveolar em ratas com 24 meses e detectamos ação importante da OT na maior formação óssea nesses animais, o que não foi observado em animais com 12 meses. Esses resultados evidenciaram a necessidade de investigação da ação da OT, visando à compreensão dos mecanismos envolvidos na maior formação óssea em organismos senis, no período caracterizado pela aciclicidade estral (estropausa – 24 meses) comparado com organismos adultos com ciclo estral regular (12 meses). Nesse estudo foi avaliada a influência da idade (12 e 24 meses) e a ação da OT associada a indutores osteogênicos clássicos durante a diferenciação em linhagem osteogênica de células-tronco mesenquimais (CTMs) a partir do estroma da medula óssea de fêmures de ratas Wistar, com 12 e 24 meses. Quatro grupos de cada idade foram formados, sendo: MC (meio de crescimento); MO (meio de crescimento suplementado com indutores osteogênicos); MCO (meio de crescimento + OT); MOO (meio de crescimento suplementado com indutores osteogênicos + OT). O ensaio com vermelho de alizarina demonstrou mineralização biológica a partir do 17º dia no grupo com 12 meses em ambos os grupos osteogênicos (MO e MOO), porém a adição de OT (MOO) promoveu a mineralização a partir do 14º dia. No grupo de 24 meses não foi detectada mineralização em nenhum dia experimental no grupo MO, porém a adição de OT no meio (MOO) promoveu a mineralização a partir do 17º dia. As escalas de expressão gênica de osterix, ocitocina e receptor de ocitocina foram menores no grupo 24 meses que no grupo 12 meses, assim como as proteínas de matriz analisadas (sialoproteína óssea, osteopontina e osteocalcina)...

Peripheral oxytocin (OT) has been shown to have direct effects on the skeleton, and to provide stimulus for the activity of osteoblasts, as an anabolic factor affecting bone mass. The effects of OT can vary with age. Here we evaluated the effects of OT on the differentiation of bone marrow mesenchymal stem cells, in 12- and 24-months-old Wistar female rats. Four treatment groups were formed for each age group: control (C) (growth medium); osteogenic medium (OM); control + oxytocin (C+OT) (growth medium + oxytocin) and osteogenic medium + oxytocin (OM+OT). Alizarin Red assay was performed and in cells from 12-month-old rats mineralization was detected on day 17 in OM. However, the addition of OT to osteogenic medium (OM+OT) provided biological mineralization on day 14. In cells from 24-month-old acyclic rats, the OM subgroup showed no mineralization; in the OM+OT subgroup mineralization was detected on day 17. Osterix, matrix protein (bone sialoprotein, osteopontin and osteocalcin) and oxytocin gene expression was lower in the 24-month-old than the 12-month-old group; these proteins were increased by OT. The oxytocin receptor was up-regulated only in the 12-month-old group and PGE2 showed an increase in the OM group than in the OM + OT group in both age groups. These findings suggest that OT shows different, differentiation-dependent actions, and is important in the biological mineralization performed by osteoblasts, mainly in cells of bone marrow mesenchymal stem cells from acyclic 24-month-old rats highlighting oxytocin as a potential therapy for control of osteoporosis