IL-6, IL-10, sFas, granulysin and indicators of intestinal permeability as early biomarkers for a fatal outcome in COVID-19.

The clinical presentation of coronavirus disease 2019 (COVID-19), which is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), ranges between mild respiratory symptoms and a severe disease that shares many of the features of sepsis. Sepsis is a deregulated response to infection that causes life-threatening organ failure. During sepsis, the intestinal epithelial cells are affected, causing an increase in intestinal permeability and allowing microbial translocation from the intestine to the circulation, which exacerbates the inflammatory response. Here we studied patients with moderate, severe and critical COVID-19 by measuring a panel of molecules representative of the innate and adaptive immune responses to SARS-CoV-2, which also reflect the presence of systemic inflammation and the state of the intestinal barrier. We found that non-surviving COVID-19 patients had higher levels of low-affinity anti-RBD IgA antibodies than surviving patients, which may be a response to increased microbial translocation. We identified sFas and granulysin, in addition to IL-6 and IL-10, as possible early biomarkers with high sensitivity (>73 %) and specificity (>51 %) to discriminate between surviving and non-surviving COVID-19 patients. Finally, we found that the microbial metabolite d-lactate and the tight junction regulator zonulin were increased in the serum of patients with severe COVID-19 and in COVID-19 patients with secondary infections, suggesting that increased intestinal permeability may be a source of secondary infections in these patients. COVID-19 patients with secondary infections had higher disease severity and mortality than patients without these infections, indicating that intestinal permeability markers could provide complementary information to the serum cytokines for the early identification of COVID-19 patients with a high risk of a fatal outcome.

Avaliação do efeito da quercetina em ratos Wistar com síndrome metabólica / Evaluation of the effect of quercetin in Wistar rats with metabolic syndrome

Introdução: O tratamento da síndrome metabólica (SM) é um desafio, uma vez que terapias não medicamentosas são de difícil implementação e o tratamento farmacológico ideal não está totalmente estabelecido. Objetivo: Avaliar o efeito da quercetina na pressão arterial (PA), dislipidemia e acúmulo de gordura visceral em modelo experimental de SM induzida por dieta hiperlipídica. Material e Métodos: Ratos Wistar receberam ração hiperlipídica a partir da quarta semana de vida, por 20 semanas. O grupo tratado recebeu quercetina a partir da oitava semana de vida. Avaliou-se semanalmente o peso corporal e a PA de cauda por pletismografia. Ao final do experimento foram realizados testes de perfil glicêmico e lipídico. Resultados:A administração de dieta hiperlipídica se associou ao desenvolvimento de SM, caracterizada por acúmulo central de gordura, hipertensão arterial, hiperglicemia e hipertrigliceridemia. A quercetina não apresentou eficácia no tratamento das comorbidades que compõem a SM. Conclusão: A administração crônica diária da quercetina em modelo experimental de SM induzida por dieta hiperlipídica não alterou de forma significante o perfil nutricional, metabólico e pressórico dos animais.

Introduction: The treatment of the metabolic syndrome (MetS) is a challenge, since nonpharmacologic therapies are difficult to implement and the ideal pharmacologic treatment has not been completely established. Aim: To evaluate the effect of quercetin in blood pressure (BP), dyslipidemia, visceral fat accumulation, in an experimental model of MetS induced by a hyperlipidic diet. Material and Methods: Wistar rats received high fat diet feed from the fourth week of life for 20 weeks. The treatment group received quercetin from the eighth week of life. Body weight and tail BP through pletysmography were evaluated weekly. At the end of the experiment, tests of glucose and lipid profile. Results: The administration of a high fat diet was associated to the development of MetS, characterized by an accumulation of central fat, arterial hypertension, hyperglycemia, and hypertriglyceridemia. Quercetin was not effective in the treatment of comorbidities associated with MetS. Conclusion: Chronic daily administration of quercetin in an experimental model of MetS induced by a hyperlipidic diet did not significantly alter the nutritional, metabolic, and pressure profile of the animals.

Evaluation of the anti-osteoporotic effect of Ginkgo biloba L. in Wistar rats with glucocorticoid-induced-osteoporosis by bone densitometry using dual-energy x-ray absorptiometry (DEXA) and mechanical testing

ABSTRACT Evaluate the effect of the extract of Ginkgo biloba in the bone alkaline phosphatase, bone mineral density, in the mechanical properties of the tibia in rats with glucocorticoid-induced-osteoporosis. After osteoporosis induction, the rats were divided into five groups: Osteoporosis; EGb1 (28 mg/Kg); EGb2 (56 mg/Kg); alendronate (0.2 mg/animal) and control. The animals were treated during 20 and 30 days. The control group was compared with the osteoporosis's (Student's t-test), while the other were analyzed by ANOVA test followed by Tukey/Dunnett'T3 (p<0.05). In the osteoporosis group the bone alkaline phosphatase, bone mineral density, the bone stiffness, the maximum load and the resilience were reduced. The bone alkaline phosphatase values increased in the EGb1 and EGb2 groups (30 days). In addition, in the EGb2 and alendronate groups (20 and 30 days) the bone mineral density increased. The extract of Ginkgo biloba restored bone alkaline phosphatase and bone mineral density using dual-energy x-ray absorptiometry.

Rats treated with Hypericum perforatum during pregnancy generate offspring with behavioral changes in adulthood

Abstract Drugs used in the treatment of depression can cross the placenta giving rise to questions regarding the effects these drugs exert on the fetus. Hypericum perforatum L., Hypericaceae, is a natural product used to treat depression. However, information about its toxicity and the occurrence of alterations in the central nervous system development of the offspring is scarce. This work assessed the behavior of adult male rats born from mothers treated with Hypericum extract during gestation and analyzed the fluorescence of the extract in different organs of mothers and fetuses. Male pups were divided into three treated groups, corresponding to the administration of the Hypericum extract to mothers at the dose levels of 36 mg/kg, 72 mg/kg and 144 mg/kg, and one control group in which the mothers received distilled water. At 90 days of age, the offspring underwent the following tests: rotarod, pentobarbital-induced sleep time, elevated plus maze, hole-board and forced swimming test. The observed fluorescence indicated the presence of the extract in all tissues analyzed. The obtained results suggest lasting changes in the performances displayed in the CNS, depression and anxiety tests, indicating that the use of Hypericum during gestation could interfere with the behavioral development of the offspring reducing anxiety and depression when they become adults. We suggest that these alterations are associated with the reprogramming of the brain regions related to changes in emotional reactivity.

Ausência de toxicidade reprodutiva e sistêmica em ratos wistar tratados com extrato seco de gingko biloba durante o período de desmame à puberdade / Absence of reproductive and sistemic toxicity in rats treated with Gingko biloba extract from weaning to puberty

Introdução: O Gingko biloba (EGb) é um fitoterápico usado há séculos, porém com poucos estudos referentes a seus efeitos sobre o período pós-natal. Estudos dessa natureza vêm sendo preconizados pela Agência Europeia de Medicina, visto que muitos órgãos completam seu desenvolvimento nesse período, inclusive o sistema reprodutor. Objetivo: Avaliar o efeito do extrato seco de EGb sobre o desenvolvimento do sistema reprodutor de ratos, tratados desde o desmame até o fim da puberdade. Métodos. Ratos Wistar foram tratados com 25mg/kg/massa corporal (EGb 25); 50 mg/kg (EGb 50) e 100 mg/kg (EGb 100). Controle (C ­ 0,1ml água destilada), por gavage dos 25 aos 45 dias de vida pós-natal. Variáveis observadas: indícios clínicos de toxicidade sistêmica, peso corporal, descida dos testículos, evolução da morfologia da glande, peso de rins, baço e fígado e dos órgãos do sistema reprodutor. Hematimetria, Concentração de hemoglobina. Concentração de espermatozoides na secreção epididimária. Resultados: Não foram encontradas diferenças significativas em quaisquer das variáveis. Conclusão: A exposição ao extrato seco de EGb durante o período pré-puberal e puberal em ratos Wistar não altera o desenvolvimento do sistema reprodutor masculino.

Introduction: Gingko biloba extract (EGb) is a phytotherapic that has been used for centuries but there is no studies concerning their effects during the postnatal period. This kind of research had been suggested by the European Medicine Agency since there are organs that complete their development in this period, including reproductive organs. Purpose: To evaluate the effect of EGb dry extract upon the rat reproductive system from weaning to 45 postnatal days. Methods: Wistar rats were treated with 25mg/kg/body weight (EGb 25); 50 mg/kg (EGb 50) and 100 mg/kg (EGb 100). Control (C 0,1ml distilled water). Variables: clinical signs of systemic toxicity, body weight, testicles descent, evolution of glans morphology, kidneys, liver, spleen and reproductive organs weights. Hematimetry. Haemoglobin concentration. Sperm concentration in the epidydimal secretion. Results: No significant differences were observed in none of the observed variables. Conclusion: The EGb dry extract exposition to prepuberal and puberal rats do not alter the reproductive system development.

Toxicological evaluation of the flavonoid rutin on the reproductive system of wistar rats / Avaliação toxicológica do flavanoide rutina no sistema reprodutor de ratos wistar

Introduction: Rutin, a flavonoid commonly found in nature, has anti-mitotic, vasoprotective, and antihyperlipidemic activity. When hydrolyzed as quercetin, it promotes inhibition of spermatozoa motility, alterations in the prostate, and in the levels of testosterone and dihydrotestosterone. Objective: This study aimed to evaluate the toxicity of rutin in Wistar rats. Methods: Animals were divided into Control (1 ml of distilled water), Treated I, II and III, respectively receiving 5, 10 and 20 mg/kg/day of rutin for seven consecutive days. When euthanasia was performed after 10, 42 and 60 days into the experiment, a laparotomy was performed and the testicles, prostate, seminal vesicles, epididymis, epididymal spermatozoa to be counted, as well as the liver, spleen and kidneys were removed. Hematological and biochemical tests were performed. Results: Hepatomegaly was observed and in the reproductive system, the weight of the epididymis was reduced, not affecting any other organ examined. Conclusion: Except by the reduction of the weight of the epididymis, which is reversible at 42 days of completion of treatment, no suggestive data of the toxicity of rutin on the reproductive system of adult rats were found.

Efeito do extrato de ginkgobiloba (EGb) sobre a toxicidade sistêmica e órgãos do sistema reprodutor masculino de ratos Wistar adultos / Effect of ginkgobiloba extract (EGb) on systemic toxicity and organs of the male reproductive system of adult Wistar rats

O Extrato de Ginkgobiloba (EGb) é um dos fitoterápicos mais consumidos no mundo. Entretanto ainda há escassez de ensaios toxicológicos em animais e o risco à exposição humana principalmente pelos compostos alquilfenóis, representados pelos ácidos ginkgólicos, que podem causar quadros alérgicos e serem compostos mutagênicos e carcinogênicos. O presente trabalho teve o objetivo de avaliar a toxicidade sistêmica do EGb. Oitenta ratos Wistar de três meses de idade foram tratados com água destilada (Grupo Controle) e extrato aquoso de Ginkgobilobanas seguintes doses: 3,5 (EGb 3,5); 7,0 (EGb 7,0) e 14,0mg/kg (EGb 14,0) uma vez ao dia, por 56 dias consecutivos. Foram avaliados semanalmente, o peso dos animais (g) e a estimativa de consumo diário de ração (g). Indícios de sinais de toxicidade sistêmica como perda de peso, piloereção, diarreia, cromodacriorreia, estereotipias, alterações da atividade locomotora e comportamentais e mortes também foram monitorados. Após anestesia, o sangue dos animais foi coletado para avaliação de hemograma completo e dosagem bioquímica de ureia, creatinina e alanina aminotransferase (ALT). Após a eutanásia, os animais foram submetidos à necropsia e os testículos esquerdo e direito, epidídimo esquerdo, vesícula seminal repleta, próstata ventral, rins esquerdo e direito, fígado e baço foram removidos e pesados em balança de precisão. Durante todo o procedimento experimental não foram observados nos animais sinais clínicos de toxicidade sistêmica e mortes. Houve diferenças estatísticas da estimativa de consumo de ração na sexta semana e oitava semanas de avaliação, embora sem diferença no peso corporal. Não houve diferença no peso dos órgãos e na análise bioquímica sérica. Na avaliação hematológica dos animais, houve diferença estatística significativa na hemoglobinometria em que o grupo EGb 14,0 apresentou-se estatisticamente superior ao grupo EGb 3,5.A concentração de hemoglobina globular média também apresentou diferença estatística significativa, em que o EGb 3,5 apresentou médias inferiores aos grupos EGb 7,0 e EGb 14,0 e o grupo controle apresentou média inferior ao grupo EGb 14,0. Sugere-se que o EGb no presente trabalho, e com as doses utilizadas, não causou toxicidade sistêmica e nem provocou alterações em órgãos de ratos Wistar.

The Ginkgobiloba Extract (EGb) is one of the most commonly consumed herbal in the world. However there are still few toxicity tests on animals and the risk of human exposure mainly by alkyl compounds, represented by acids, which can cause allergies and are mutagenic and carcinogenic compounds. This study had the objective of evaluate the systemic toxicity of EGb. Eighty Wistar rats, three months of age were treated with distilled water (Control Group) and aqueous extract Ginkgobilobanas following doses: 3.5 (EGb 3.5); 7.0 (EGb 7.0) and 14,0mg / kg (14.0 EGb) once a day for consecutive 56 days. Were evaluated weekly animal weight (g) and the estimated daily intake (g). Evidence of systemic signs of toxicity such as weight loss, piloerection, diarrhea, stereotypies and behavioral changes in motor activity and deaths were also monitored. After anesthesia, the animals were collected for evaluation of complete blood count and biochemical analysis of urea, creatinine and alanine aminotransferase (ALT). After euthanasia, the animals were autopsied and the left and right testis, left epididymis, seminal vesicle filled, ventral prostate, left and right kidneys, liver and spleen were removed and weighed on a precision scale. Throughout the experimental procedure were not observed in animals clinical signs of systemic toxicity and deaths. Were no statistical differences in the estimate of feed intake in the sixth week and eighth week evaluation, although no difference in body weight. There were no differences in organ weight and serum biochemical analysis. Hematological evaluation of the animals, there was a statistically significant difference in Hemoglobinometry where 14.0 EGb group was statistically higher than the EGb group 3,5. A mean corpuscular hemoglobin concentration also showed a statistically significant difference in the EGb 3 5 showed an average lower than 7.0 and EGb groups EGb 14.0 and the control group showed less than 14.0 EGb group. It is suggested that EGb in this work, and the doses used, did not cause systemic toxicity nor caused changes in organs of Wistar rats.

Evaluation of postpartum behaviour in rats treated with Hypericum perforatum during gestation

Gestational depression is detrimental to the health of the mother and the offspring and contributes to the appearance of depressive and anxiety symptoms during the postnatal period. Traditional antidepressants have undesirable side effects when utilised during gestation, but Hypericum perforatum has been characterised as an efficient and safe antidepressant that prevents the recurrence of symptoms. This study verified the effects of Hypericum perforatum on the behaviour of Wistar rats that were treated during gestation and evaluated 10 and 60 days post-treatment. Pregnant Wistar rats were divided into four groups of ten animals each: one control group that received distilled water and three treatment groups that were treated orally with 36, 72 or 144 mg/kg Hypericum perforatum extract. At 10 and 60 days after parturition and post-treatment, the rats were submitted to the holeboard, the tail suspension, and the forced swim tests. The animals treated with 144 mg/kg Hypericum perforatum exhibited greater head-dipping activity in the hole-board test and reduced immobility in the tail suspension and forced swim tests, suggesting less anxiety and depression 10 and 60 days post-treatment.The results indicated that treating rats with Hypericum perforatum during the gestational period decreased depressive behaviour and anxiety 10 and 60 days post-treatment.

The effect of the Ginkgo biloba extract in the expression of Bax, Bcl-2 and bone mineral content of Wistar rats with glucocorticoid-induced osteoporosis.

OBJECTIVE: Evaluate the effects of the extract of Ginkgo biloba (EGb) in the glucocorticoid-induced-osteoporosis through the Bax and Bcl-2 expressions by osteoblast cells, the x-ray and bone density of the tibia. METHOD: Rats were divided into five groups: osteoporosis; EGb1 (28 mg/kg); EGb2 (56 mg/kg); alendronate (0.2 mg/animal) and control. The treatments were conducted for 20 (n = 30) and 30 days (n = 30). The Bax and Bcl-2 expressions were evaluated in osteoblasts of the mandibular alveolar bone. The tibias were radiographed to evaluate the X-ray and bone density. The control group was compared with the osteoporosis' (Student's t-test/Mann-Whitney). The other groups were analyzed by analysis of variance test followed by Dunnett/Dunnett T3 (p < 0.05). RESULTS: When compared the osteoporosis to the control group (p <0.05): Bax and x-ray density increased; Bcl-2 and the bone density reduced. When compared with the osteoporosis group (p < 0.05), alendronate (30 days), EGb1 and EGb2 (20/30 days) increased the Bcl-2 expression; EGb2 and alendronate (20 days) EGb1 and EGb2 (30 days) reduced the Bax expression; and EGb1 and EGb2 (20/30 days) reduced the X-ray density. CONCLUSIONS: The EGb improved the Bcl-2 and reduced the Bax expression by osteoblasts in the mandibular alveolar bone and recovered the mineral content in the tibia of rats with glucocorticoid-induced-osteoporosis.

Effects of a continuous-combined regimen of low-dose hormone therapy (oestradiol and norethindrone acetate) and tibolone on the quality of life in symptomatic postmenopausal women: a double-blind, randomised study.

OBJECTIVE: This study compared the effects of a continuous-combined regimen of low-dose hormone therapy (LD-HT) versus tibolone and supplemental calcium/vitamin D3 (control) on quality of life (QoL) in symptomatic postmenopausal women. DESIGN: This study was a prospective, randomised, double-blind, comparative trial with a control group. SETTING: The study was conducted in a climacteric outpatient clinic in the University Hospital of Federal University of Juiz de Fora, Brazil. POPULATION: A total of 174 postmenopausal women under 60 years of age who attended the climacteric outpatient clinic between June 2009 and June 2011 were recruited. These women complained of moderate or intense vasomotor symptoms and exhibited no contraindications for the use of hormone therapy. INTERVENTIONS: The patients were randomised into three groups: (1) daily treatment with 2.5mg tibolone (n=64), (2) 50mg calcium carbonate+200 IU vitamin D3 (Ca/Vit D3, n=54) or (3) 1mg oestradiol+0.5mg norethindrone acetate (E2/NETA, n=56) for 12 weeks. PRIMARY OUTCOME MEASURES: The primary outcome was the evaluation of QoL using the Women's Health Questionnaire (WHQ) in all subjects at baseline and after 4, 8 and 12 weeks of treatment. RESULTS: A total of 130 women in the following groups completed the study: tibolone (n=42), Ca/Vit D3 (n=44) and E2/NETA (n=44). An improved QoL based on the WHQ was observed at T0 (80.12±14.04, 77.73±15.3, 77.45±15.4) and T12 (57.0±15.5, 55.7±16.7, 58.4±12.6) for the tibolone, E2+NETA and Ca/Vit D3 groups, respectively (p values <0.05). The three groups exhibited significantly different scores at T12 for sexual behaviour and vasomotor symptoms. The tibolone group exhibited better sexual function compared with the E2/NETA and Ca/Vit D3 groups (4.2±26, 5.6±2.8, 5.4±2.8, respectively, p values <0.05). LD-HT was superior to tibolone and Ca/Vit D3 treatment for improvements in vasomotor symptoms (3.2±1.5, 4.0±1.8, 4.3±2.0, respectively, p values <0.05). Adverse effects were few and mild. CONCLUSIONS: An improved QoL was observed in the three study groups. Tibolone primarily improved sexual function, and E2/NETA exhibited a superior response for vasomotor symptoms.

Dissociação da resposta anti-hipertensiva e metabólica à losartana e espironolactona em ratos com síndrome metabólica experimental / Dissociation of antihypertensive and metabolic response to losartan and spironolactone in experimental rats with metabolic sindrome

INTRODUÇÃO: O tratamento da hipertensão arterial (HA) em indivíduos com síndrome metabólica (SM) é um desafio, uma vez que terapias não medicamentosas são de difícil implementação e o tratamento farmacológico ideal não está totalmente estabelecido. OBJETIVO: Avaliar o bloqueio do sistema renina angiotensina aldosterona (SRAA) na pressão arterial (PA), na função e na morfologia renais em modelo experimental de SM induzida por dieta hiperlipídica. MÉTODOS: Ratos Wistar receberam ração hiperlipídica a partir da quarta semana de vida, por 20 semanas. Os grupos tratados receberam Losartana ou Espironolactona a partir da oitava semana de vida. Avaliou-se semanalmente o peso corporal e a PA de cauda por pletismografia. Ao final do experimento, foram realizados testes de tolerância oral à glicose, perfil lipídico, clearance de creatinina, medida direta da PA, análise morfométrica renal. RESULTADOS: A administração de dieta hiperlipídica se associou ao desenvolvimento de SM, caracterizada por acúmulo central de gordura, hipertensão arterial, hiperglicemia e hipertrigliceridemia. Nesse modelo não foram observadas alterações da histomorfometria renal. O bloqueio do receptor AT1 da angiotensina II (Ang II) preveniu o desenvolvimento da HA. O bloqueio mineralocorticoide não apresentou eficácia anti-hipertensiva, porém, associou-se à redução da gordura abdominal. CONCLUSÃO: A dissociação da resposta anti-hipertensiva aos bloqueios dos receptores da Ang II e mineralocorticoide indica a participação da Ang II na gênese da HA associada à obesidade. A redução da obesidade central com a Espironolactona sugere a presença de efeito adipogênico mineralocorticoide.

INTRODUCTION: The treatment of arterial hypertension (AH) in patients with metabolic syndrome (MS) is a challenge, since non drug therapies are difficult to implement and optimal pharmacological treatment is not fully established. OBJECTIVE: To assess the blockade of the rennin angiotensin aldosterone system (RAAS) in blood pressure (BP) in renal function and morphology in an experimental model of MS induced by high fat diet. METHODS: Wistar rats were fed on high fat diet from the fourth week of life, for 20 weeks. The groups received Losartan or Spironolactone from the eighth week of life. We weekly evaluated the body weight and BP by tail plethysmography. At the end of the experiment oral glucose tolerance, lipid profile, creatinine clearance tests, and the direct measurement of BP were performed. A morphometric kidney analysis was performed. RESULTS: The administration of high-fat diet was associated with the development of MS, characterized by central fat accumulation, hypertension, hyperglycemia and hypertriglyceridemia. In this model there were no changes in renal histomorphometry. The blockade of angiotensin II (Ang II) receptor AT1 prevented the development of hypertension. The mineralocorticoid blockage did not have antihypertensive efficacy but was associated with reduction of abdominal fat. CONCLUSION: The dissociation of the antihypertensive response to the blockades of Ang II receptors and mineralocorticoid indicates the involvement of Ang II in the pathogenesis of hypertension associated with obesity. Reduction of central obesity with Spironolactone suggests the presence of mineralocorticoid adipogenic effect.

Efeito da ipriflavona sobre ratas Wistar e suas ninhadas / Effect of ipriflavone on Wistar rats and their litters

OBJETIVO: Avaliar os efeitos da ipriflavona durante a fetogênese, já que não foram encontrados estudos visando avaliar seu efeito durante este período. MÉTODOS: Foram utilizadas 60 ratas prenhes, distribuídas aleatoriamente em quatro grupos (n=15). G-controle (1 mL de água destilada) e três grupos tratados com ipriflavona, via intragástrica, do 16º ao 20º dia pós-coito (PC): G-300 (300 mg/kg), G-1500 (1.500 mg/kg) e G-3000 (3.000 mg/kg). Os animais foram pesados e sacrificados no 21º dia por exsanguinação total sob anestesia (xilazina (10 mg/kg) e quetamina (90 mg/kg) via intraperitoneal. Foi realizado hemograma completo e dosagens séricas de colesterol, triglicérides, AST, ALT, ureia, creatinina e glicose das ratas prenhes. Após laparotomia foram removidos e pesados fígado, rim, suprarrenais, baço e ovários; os fetos e placentas foram pesados obtendo-se o peso médio das ninhadas. Quatro fetos (dois machos e duas fêmeas) por mãe foram aleatoriamente designados para obter-se o comprimento e peso de cérebro, fígado, rins e pulmões. Para a análise estatística utilizou-se o teste ANOVA seguido do teste de Dunnet; para dados não homocedásticos e sem distribuição normal, foi usado o teste de Kruskal-Wallis, seguido de Mann-Whitney; as proporções foram analisadas pelo teste do χ² (p<0,05) RESULTADOS: Níveis de triglicérides (mg/dL): G-Controle (138,8±21,8); G-300 (211,2±63,9); G-1500 (251,5±65,2); G-3000 (217,7±49,6); p<0.05. Peso corporal dos fetos (g): G-Controle (machos 3,3±0,3; fêmeas 3,1±0,3); G-300 (machos 3,4±0,2; fêmeas 3,1±0,4); G-1500 (machos 3,5±0,3; fêmeas 3,2±0,3); G-3000 (machos 3,4±0,5; fêmeas 3,1±0,4). CONCLUSÃO: A ipriflavona não causou toxicidade materna, mas elevou níveis de triglicérides e reduziu o hematócrito em doses elevadas, o tamanho, peso corporal e de órgãos fetais não foram alterados. Não foram observadas malformações externas nem mortes fetais.

PURPOSE: Evaluate the effects of ipriflavone during fetogenesis, since no studies have been conducted to assess its effect during this period. METHODS: 60 pregnant rats were divided randomly into four groups (n=15). G-control (1 mL of distilled water) and three groups treated intragastrically with ipriflavone from the 16th to the 20th post coitus (PC) day: G-300 (300 mg/kg), G-1,500 (1,500 mg/kg) and G-3,000 (3,000 mg/kg). The animals were weighed, anaesthetized intraperitoneally with xylazine and ketamine at doses of 180 mg/kg and 10 mg/kg, respectively, and sacrificed by total exsanguination on the 21st day. A complete blood count was performed and serum cholesterol, triglycerides, AST, ALT, urea, creatinine, and glucose were determined in pregnant rats. After laparotomy, the liver, kidneys, adrenals, spleen and ovaries were removed and weighed; fetuses and placentas were also weighed to obtain the average weight of the litters. Four fetuses (two males and two females) were chosen at random for the determination of the length and weight of brain, liver, kidneys and lungs. Statistical analysis: ANOVA followed by Dunnett's test. For raw data without normal distribution and homoscedasticity, we used the Kruskal-Wallis test followed by the Mann-Whitney test. Proportions were analyzed by the χ² test (p<0.05). RESULTS: Triglyceride levels (mg/dL) were: Control-G (138.8±21.8), G-300 (211.2±63.9) G-1,500 (251.5±65.2) G-3,000 (217.7±49.6); p<0.05. The body weight of fetuses (g) was: G-Control (male 3.3±0.3; female 3.1±0.3), G-300 (male 3.4±0.2; female 3.1±0.4), G-1,500 (male 3.5±0.3; female 3.2±0.3), G-3,000 (male 3.4±0.5; female 3.1±0.4). CONCLUSION: Ipriflavone did not cause maternal toxicity, but increased triglyceride levels and reduced hematocrit at higher doses. The body and organ weights of the fetuses did not change with dam treatment. There were no external malformations or fetal deaths.

Efficacy of Morus nigra L. on reproduction in female Wistar rats.

Morus nigra L. is a plant employed as a substitute for the conventional hormonal replacement therapy. This work analyzes the estrogenic effect of M. nigra on the reproductive system and embryonic development of Wistar rats. Female rats were orally treated with M. nigra hydroalcoholic extract (MnHE) at the dose levels of 25, 50, 75, 350 and 700 mg/kg of body weight over 15 days, and continued through mating until the 14th day of gestation. Vaginal smears were performed daily and the body weight of the females was recorded at 5 days intervals. On day 15 of gestation, the females were killed and their kidneys, liver, spleen and ovaries were removed and weighed. The number of implants, resorptions, and live and dead fetuses were evaluated. Histological sections of ovaries, measurement of the height of the uterine epithelium and vaginal smears were performed to assess the estrogenic activity. The results showed that the administration of MnHE did not significantly alter the analyzed variables. Therefore, considering the experimental model used in this study, the data obtained indicate that M. nigra did not exhibit any estrogenic activity nor did exert a toxic effect on the female reproductive system and on the embryonic development of rats.

Avaliação do potencial interceptivo da ipriflavona em ratas wist / Evaluation of the potential of interceptive ipriflavone in wistar rats

A ipriflavona possui efeito inibitório sobre o citocromo p450 e sobre a proliferação do DNA, interferindo na síntese de hormônios estereoidianos, e consequente interferência no transporte e desenvolvimento pré-implantacional e na implantação do blastocisto. Consumidos em altas doses pode acarretar abortamentos e malformações. Para verificar a embriotoxicidade da ipriflavona durante as fases de transporte tubário e implantação do blastocisto de ratas, ratas Wistar prenhes foram tratadas duas vezes ao dia com 1mL de suspensão aquosa de ipriflavona, nas doses de 0 (C), 300 (T1), 1500 (T2) e 3000 (T3) mg/kg/dose, durante os oito primeiros dias de prenhez com eutanásia no 14o dia. Indícios de toxicidade foram avaliados por análises hematimétricas, bioquímicas e comportamentais. Foram contados fetos vivos, mortos e reabsorvidos (inicial e tardiamente). Os animais não apresentaram sinais clínicos de toxicidade. Índice de implantação e perdas pré-embrionária sofreram interferência do uso da ipriflavona. Dados apontam para um efeito estrogênico da ipriflavona, observados na interferência da implantação do blastocisto de ratas Wistar.

Ipriflavone has inhibitory effect on the cytochrome p450 and the proliferation of DNA, interfering with hormone synthesis estereoidianos, and consequent interference with the transport and preimplantation development and implantation of the blastocyst. Consumed in high doses can cause miscarriages and malformations. To verify the embryotoxicity of ipriflavone during the phases of tubal transport and implantation of the blastocyst in rats, Wistar rats were treated twice daily with 1 mL of aqueous suspension of ipriflavone at doses of 0 (C), 300 (T1), 1500 (T2) and 3000 (T3) mg / kg / dose during the first eight days of pregnancy with euthanasia on day 14. Signs of toxicity were characterized by hematological, biochemical and behavioral. Live fetuses were counted, dead and resorbed (early and late). The animals showed no clinical signs of toxicity. Index pre-implantation embryo and losses suffered interference from the use of ipriflavone. Data point to an estrogenic effect of ipriflavone, due to interference in blastocyst implantation in Wistar rats.

Preliminary assessement of the estrogenic potential of Apuleia leiocarpa (Vogel) J.F. Macbr., Fabaceae, Platypodium elegans Vogel, Fabaceae, and Brosimum guianense (Aubl.) Huber, Moraceae, on the wistar rat reproductive system / Avaliação preliminar do potencial estrogênico de Apuleia leiocarpa (Vogel) J.F. Macbr., Fabaceae, Platypodium elegans Vogel, Fabaceae, and Brosimum guianense (Aubl.) Huber, Moraceae, no sistema reprodutor de ratos Wistar

A group of primates (Alouatta guariba) was studied in its natural habitat, where a drastic populational reduction was detected. It is suspected that this reduction is due to the inhibition of fertility caused by the consumption of Apuleia leiocarpa (Vogel) J.F. Macbr., Fabaceae, Platypodium elegans Vogel, Fabaceae, and Brosimum guianense (Aubl.) Huber, Moraceae. These plants are reported to have cumarins, which have been shown to affect ovarian follicular development in rats. This work investigates the estrogenic activity of these plants on the uterus and vagina using castrated rats as the biological model. Pubescent castrated rats were treated for five days with A. leiocarpa, P. elegans and B. guianense hydroalcoholic extracts (50 mg/rat). Uterus and pituitary gland weight, vaginal cornification and opening were evaluated. The results showed that the administration of the extracts did not significantly alter the parameters analyzed. This preliminary investigation indirectly indicates the absence of estrogenic effect on the rat reproductive system and no relation to the populational reduction of this particular group of primates.

Um grupo de primatas da espécie Alouatta guariba foi estudado em seu habitat natural na Mata Atlântica, onde foi observada uma drástica redução populacional dessa espécie. Suspeita-se que essa redução se deve à inibição da fertilidade das fêmeas devido ao consumo de Apuleia leiocarpa (Vogel) J.F. Macbr., Fabaceae, Platypodium elegans Vogel, Fabaceae e Brosimum guianense (Aubl.) Huber, Moraceae. Estudos fitoquímicos indicaram a presença de cumarinas, especialmente em B. guianense e P. elegans, cujo efeito adverso no desenvolvimento de folículos ovarianos foi previamente relatado em ratas. Este trabalho investiga a atividade estrogênica dessas plantas no útero e vagina utilizando ratas castradas como modelo experimental. Ratas Wistar pubescentes castradas foram tratadas por cinco dias com os extratos hidroalcoólicos de A. leiocarpa, P. elegans and B. guianense (50 mg/rata). Foram analisados os seguintes parâmetros: peso de útero e hipófise, cornificação e abertura vaginal. Os resultados preliminares obtidos mostraram que a administração dos extratos não alterou significativamente as variáveis analisadas, indicando, indiretamente, a ausência de efeito estrogênico no sistema reprodutor das ratas tratadas com as plantas citadas. Esses dados sugerem que o consumo dessas plantas não está relacionado com a redução populacional observada no grupo de primatas da espécie A. guariba.

Effect of interventions with ingestion of legumes and/or supervised exercise on the lipid profile of young, healthy sedentary women / Efecto de las intervenciones con ingesta de leguminosas y/o ejercicio supervisado sobre el perfil lipídico de mujeres jóvenes, sanas y sedentarias

Objective: To contribute to the knowledge of some aspects of the Healthy Life Style by studying the effects of including legumes in the diet and exercise at two intensity levels, along with the lipid profile of young sedentary women living at 2640 meters above sea level. Materials and methods: The study included a non-randomized clinical trial with four intervention groups: exercise at 45% VO2 peak plus legumes in diet, exercise at 65% VO2 peak plus legumes in diet, only exercise at 65% VO2 peak, and only inclusion of legumes in diet. In each group, 20 to 23 sedentary women were included. The intervention was carried out for four weeks, three days a week. Exercise prescription was based on measurement of VO2 peak by ergospirometry; and the current intervention was monitored with heart-rate monitors. The outcome variables were total serum cholesterol, LDL cholesterol, HDL cholesterol, and triacylglycerols measured at baseline and after two and four weeks of intervention. Results: The measurements taken of participants in the group of exercise at 65% VO2 peak plus legumes in diet showed a reduction of 19.8 mg/dl in total cholesterol, of 21.8 mg/dl of LDL cholesterol, of 20.7 mg/dl of tracylglycerol, and an increase of 6.2 mg/dl of HDL cholesterol. The serum levels of HDL also increased in the group of only exercise at 65% VO2 peak. No significant changes in serum levels were documented for participants in the group with only dietary modifications. Conclusions: The results suggest that different interventions that meet some of the criteria for healthy eating and life style show different effects with regards to the level of change in the lipid profile components.

Objetivo: Contribuir al conocimiento de algunos aspectos prácticos para llevar a cabo una alimentación y estilo de vida saludable, mediante el estudio de los efectos en el perfil lipídico de la inclusión de leguminosas (fríjol, lenteja y garbanzos) en la dieta de mujeres sanas pero sedentarias, acompañada de ejercicio con dos niveles de intensidad, moderada y alta, efectuados a una altura de 2,640 metros sobre el nivel del mar (msnm). Material y métodos: Estudio de intervención clínica no aleatorizada con cuatro grupos de intervención: ejercicio a 45%VO2 pico + leguminosas, ejercicio a 65%VO2 pico + leguminosas, sólo ejercicio a 45%VO2 pico y sólo inclusión de leguminosas en la dieta. En cada grupo se incluyeron de 20 a 23 mujeres jóvenes sedentarias evaluadas mediante el International Physical Activity Questionnaire (IPAQ), y la intervención se llevó a cabo durante 4 semanas. La prescripción del ejercicio se hizo previa determinación del VO2 pico por ergo espirometría, y su realización se monitoreó con sensores de frecuencia cardíaca. El efecto de la intervención se evaluó mediante los cambios en los niveles séricos de colesterol total (CT), colesterol LDL (cLDL), colesterol HDL (cHDL) y triacilgliceroles (TAG), tomados a las semanas cero, dos y cuatro. Resultados: En el grupo ®ejercicio a 65% del VO2 pico, más inclusión de leguminosas en la dieta¼ se obtuvieron los siguientes resultados significantes: una reducción promedio del colesterol total de 19.8 mg/dl (p=0.0015), del colesterol LDL de 21.8 mg/dl (p=0.0001), un aumento de colesterol HDL de 6.2 mg/dl (p=0.0001) y una disminución de los triacilgliceroles de 20.7 mg/dl (p=0.0001). En el grupo de sólo ejercicio a 65% del VO2 pico se obtuvo un aumento del HDL. No hubo modificaciones al perfil lipídico en el grupo correspondiente a sólo inclusión de leguminosas en la alimentación...

Avaliação dos parâmetros hematológicos e bioquímicos de ratas no segundo terço da gestação submetidas à ação do extrato metanólico de Cereus jamacaru DC., Cactaceae / Evaluation hematological and biochemical parameters of rats in the second thirds of the gestation submitted to the action of the extract methanol of Cereus jamacaru DC., Cactaceae

Os efeitos da administração oral do extrato metanólico (EM) de Cereus jamacaru DC., Cactaceae, foram investigados sobre os parâmetros hematológicos e bioquímicos em ratas Wistar adultas grávidas. Vinte ratas (n = 5 por grupo) foram tratadas durante quatro dias consecutivos com EM por via oral nas doses de 100, 250 e 500 mg/kg de peso e, em seguida, determinados os perfis bioquímico e hematológico. Os resultados mostraram que durante o período do tratamento não foi observado efeitos nocivos ou óbitos. Os parâmetros hematológicos e bioquímicos não foram modificados pela administração oral do EM, excetuando-se o aumento significativo de 45,7 por cento e de 41,9 por cento para alanina transferase (ALT) nas doses de 250 e 500 mg/kg além do aumento significativo, respectivo de 48,2 por cento, 39,8 por cento e 41,8 por cento para aspartato aminotransferase (AST). Em relação aos valores hematológicos, registrou-se flutuação dentro dos valores de referência na contagem diferencial de hemoglobina, de neutrófilo e de linfócito. Dessa forma, a administração do extrato metanólico de C. jamacaru não apresentou reações tóxicas sobre a maioria dos parâmetros hematológicos e bioquímicos estudados em ratas Wistar adultas grávidas. Entretanto, o aumento dos níveis séricos de AST e ALT em doses elevadas sugere uma sobrecargas hepática, as quais devem ser investigadas em maiores detalhes.

The effects of oral administration of methanol extract (ME) prepared from stems of Cereus jamacaru DC., Cactaceae were investigated on the biochemical and hematological parameters in pregnant adult Wistar rats. Twenty rats (n = 5 per group) have been treated orally for four consecutive days with ME in doses of 100, 250 and 500 mg/kg weight, and then, it was determined the biochemical and hematological profiles. The results showed that during the period of treatment there was no signs of toxicity or death. The hematological and biochemical parameters were not modified by oral administration of ME, except for a significant increase of 45.7 percent and 41.9 percent for alanine transaminase (ALT) in doses of 250 and 500 mg/kg in addition to the significant increase, of to 48.2 percent, 39.8 percent and 41.8 percent for aspartate transaminase (AST). In hematology, it was registered a fluctuation within the reference values of hemoglobin in the differential count of neutrophil and lymphocyte. In this way the administration of methanol extract of C. jamacaru does not produce toxic effects or alters the majority of biochemical and hematological studies in pregnant adult Wistar rats. However, the increase of serum ALT and AST in high doses suggests a liver overload, which must be investigated in more detail.

Avaliação hematológica e bioquímica em ratas prenhes tratadas com extrato de ginkgo bilob / Hematologic and biochemistry evaluation on pregnant rat treated with gingko biloba extract

Introdução: O extrato de Gingko biloba (GBE) é um fitoterápico usado no tratamento de doenças degenerativas e em estudos recentes tem sido demonstrado efeito nefro e hepatoprotetor de seus componentes. Material e métodos: No presente estudo, 120 ratas Wistar prenhes foram distribuídas em dois grupos experimentais ­ GB 15 e GB 21 ­ tratadas, respectivamente, do primeiro ao oitavo dia da prenhez e do oitavo ao vigésimo dia com 0, 3,5; 7,0 ou 14mg/kg/dia de extrato aquoso de GBE, via gavagem. Os animais foram eutanaziados por exsanguinação total, sob anestesia, no 15º dia ou no 20º dia de prenhez. Os seguintes parâmetros foram avaliados no sangue coletado: eritrograma, leucograma, dosagens séricas de ureia, creatinina, ALT, AST, colesterol e triglicérides. Resultados: Não foram encontradas alterações significativas no padrão hematológico de ratas tratadas nos grupos GB 15 e GB 21. Em relação ao perfil bioquímico, o grupo GB 15, tratado com as doses de 7 e 14mg/kg, evidenciou aumento dos níveis de colesterol e redução de ALT, ureia e creatinina. No grupo GB 21, tratado com as mesmas doses, não se observou aumento de colesterol, mas sim de ureia, enquanto que ALT e creatinina reduziram-se da mesma maneira que no grupo GB 15. Conclusões: Os resultados sugerem que o GBE não altera os padrões hematológicos, porém, no início da gestação aumenta os níveis de colesterol, enquanto que no final da gestação não altera o colesterol, aumentando a ureia, e durante os dois períodos de gestação reduz creatinina e ALT, o que parece confirmar os efeitos nefro e hepatoprotetor.

Introduction: The Ginkgo biloba extract (GBE) is a phytotherapic used in the treatment of neurodegenerative diseases and recent studies have demonstrated nephro and hepatoprotector effects of its components. Material and methods: In this study 120 pregnant Wistar rats were distributed among two experimental groups - GB15 e GB21 ­ treated respectively from the first to eight day of pregnancy and to the eight to de 20th day, with zero, 3.5, 7 or 14mg/kg/day of aqueous extract of Gingko biloba by gavagem. Animals were euthanized by exsanguinations under anesthesia on 15th or 21th pregnancy day. The following parameters were analyzed in the blood hemogram, hematocrit, hemoglobin, total leukocytes, cholesterol, triglycerides, urea, creatinine, aspartato aminotransferase (AST/TGO), alanina aminotransferase (ALT/TGP). Results: No hematological alteration was observed in either group. With respect to biochemistry profile the GB15, group treated with 7 and 14mg/kg, showed higher level of cholesterol and lower level of ALT, urea and creatinin. In the group GB21, treated with the same dose, there was no cholesterol alteration but higher level of urea whereas ALT and creatinin where lower than control as in GB15 group. Conclusions: GBE seems do not alter hematological profile but at early gestation increase the cholesterol level. At latter gestation do not alter cholesterol but increase urea levels. At all period of the gestation the GBE decrease creatinine and ALT seems to confirm possible nepro and hepatic protector effect.

Efeito do extrato de Ginkgo biloba L., Ginkgoaceae, na osteoporose induzida em ratas Wistar / Effect of the aqueous extract of Ginkgo biloba L., Ginkgoaceae, in induced osteoporosis in Wistar rats

Este trabalho investigou os efeitos do tratamento por vinte dias com extrato de Ginkgo biloba (EGb) na osteoporose induzida por glicocorticóides. Foram utilizadas 36 ratas divididas em seis grupos (n=6): Controle, osteoporose, controle positivo, EGb1 (14 mg EGb/mg/kg/dia), EGb2 (28 mg EGb/kg/dia) e EGb3 (56 mg EGb/kg/dia). Os tratamentos foram realizados por vinte dias, após a indução da osteoporose. Após a eutanásia foram removidos o fêmur e a mandíbula de todos os animais. A mandíbula esquerda foi radiografada digitalmente para avaliação da cortical e do suporte ósseo periodontal (SOP). A análise histomorfométrica foi realizada no fêmur e mandíbula direitos. O grupo controle foi comparado ao grupo osteoporose (Teste t de Student) e os demais grupos foram submetidos a ANOVA, seguido do teste post-hoc de Dunnett. Houve redução significava do SOP mesial, percentual ósseo alveolar (POA) mandibular, percentual ósseo trabecular (POT) do fêmur no grupo osteoporose. Houve aumento do SOP mesial no grupo controle positivo, EGb2 e EGb3. O POA da mandíbula e o POT do fêmur aumentaram nos grupos EGb2 e EGb3. O EGb nas doses de 28 mg/kg e 56 mg/kg recuperou de forma significativa o SOP mesial, o POA da mandíbula e o POT do fêmur.

The objective of this study was to investigate the effect of a 20 day treatment with extract of Ginkgo biloba (EGb) in glucocorticoid-induced-osteoporosis. 36 rats were divided into six groups (n=6): control, osteoporosis, positive control, EGb1 (14 mg EGb/kg/day), EGb2 (28 mg EGb/kg/day) and EGb3 (56 mg EGb/kg/day). Treatments were conducted for twenty days, after osteoporosis was induced. Following euthanasia the femur and mandible of all animals were removed. The left mandible was radiographed to evaluate the cortical and the periodontal bone support (PBS). The histomorphometric analysis was performed on the right mandible and the right femur. The control group was compared with the osteoporosis group (Student's t-test). The other groups were analyzed through ANOVA test followed by Dunnett post-hoc test. There was a significantly reduction in the mesial PBS, in the percentage of the alveolar bone (PAB) of the mandible and percentage of the trabecular bone (PTB) of the femur in the osteoporosis group. There was an increase in the mesial PBS in the positive control group, EGb2 and EGb3. The PAB of the mandible and the PTB of the femur increased in the EGb2 and EGb3 groups. The EGb in the 28 mg/kg and 56 mg/kg doses were effective to increase the mesial PBS, the PAB of the mandible and the PTB of the femur.

Considerações e questionamentos sobre as evidências da ovogênese pós-natal em mamíferos / Considerations and questionaments about evidence of post-natal ovogenesis in mammalian

É apresentada uma revisão de literatura sobre a origem dos ovócitos primários em ovários de fêmeas adultas de mamíferos. Apesar de ser considerado um processo exclusivamente embrionário, vem se questionando a produção dessas células germinativas após o nascimento, processo denominado neo-ovogênese. Diferentes estudos vêm sendo apresentados para demonstrar a ocorrência da nova teoria, como a existência de células-tronco germinativas no epitélio do ovário ou de células-tronco de medula óssea. Dessa forma, é importante o estudo da neo-ovogênese, já que a mesma poderá proporcionar um benefício inigualável para a área de reprodução assistida.

A literature review about the origin of primary oocytes in ovaries of female adult mammals is shown. Although it was considered only an embryonic process, the production of the germ cells after birth has been questionated, a process called neoovogenesis... Different studies have been shown to prove the occurrence of this new theory, like the existence of germ stem cells in ovarian germ epithelial or stem cells from bone marrow. Like this, it is important to study the neoovogenesis, since it may provide a unique benefit to the area of assisted reproduction.