Progesterone receptors in extratesticular ducts of the Amazonian stingray Potamotrygon wallacei: A potential role in sperm maturation and aggregate formation.

In cururu stingray (Potamotrygon wallacei Carvalho, Rosa and Araújo 2016) males, plasma progesterone (P4) levels appear to be associated with spermiation events. However, the specific contribution of P4 in sperm maturation via extratesticular ducts in this stingray species is unknown. With the aim of filling this knowledge gap, this study examined the morphology and the presence of progesterone receptors (PR) in the ducts, and analyzed the relationship of progesterone (P4) with sperm maturation and formation of aggregates. Morphological analysis showed that a columnar pseudostratified epithelium with stereocilia lined all the attached ducts. In active males, the secretory cells of the epididymis and the Leydig glands presented PR; however, these receptors were not found in the distal region of the epididymis (essential for nurturing and capacitation events) of regressing males. In the seminal vesicles of active males, the spermatozoa are parallelly aligned and embedded in a matrix to form the spermatozeugmata. The matrixes are formed by proteins secreted by the ducts and Sertoli cell cytoplasts. These structures presented PR, which suggests that P4 engages in sperm metabolism during storage. Our findings allude to the potential role of P4 in regulating the development and function of the attached ducts in different reproductive phases. Furthermore, P4 seems to be an essential component for regulating sperm progress, protein secretion, aggregate formation, and maintenance of sperm during storage in this freshwater stingray.

Diagnostic test accuracy of novel biomarkers for lupus nephritis-An overview of systematic reviews.

INTRODUCTION: Systemic lupus erythematosus (SLE) is a chronic autoimmune disease with multiorgan inflammatory involvement and a mortality rate that is 2.6-fold higher than individuals of the same age and sex in the general population. Approximately 50% of patients with SLE develop renal impairment (lupus nephritis). Delayed diagnosis of lupus nephritis is associated with a higher risk of progression to end-stage renal disease, the need for replacement therapy, and mortality. The initial clinical manifestations of lupus nephritis are often discrete or absent and are usually detected through complementary tests. Although widely used in clinical practice, their accuracy is limited. A great scientific effort has been exerted towards searching for new, more sensitive, and specific biomarkers in recent years. Some systematic reviews have individually evaluated new serum and urinary biomarkers tested in patients with lupus nephritis. This overview aimed to summarize systematic reviews on the accuracy of novel serum and urinary biomarkers for diagnosing lupus nephritis in patients with SLE, discussing how our results can guide the clinical management of the disease and the direction of research in this area. METHODS: The research question is "What is the accuracy of the new serum and urinary biomarkers studied for the diagnosis of LN in patients with SLE?". We searched for systematic reviews of observational studies evaluating the diagnostic accuracy of new serum or urinary biomarkers of lupus nephritis. The following databases were included: PubMed, EMBASE, BIREME/LILACS, Scopus, Web of Science, and Cochrane, including gray literature found via Google Scholar and PROQUEST. Two authors assessed the reviews for inclusion, data extraction, and assessment of the risk of bias (ROBIS tool). RESULTS: Ten SRs on the diagnostic accuracy of new serum and urinary BMs in LN were selected. The SRs evaluated 7 distinct BMs: (a) antibodies (anti-Sm, anti-RNP, and anti-C1q), (b) cytokines (TWEAK and MCP-1), (c) a chemokine (IP-10), and (d) an acute phase glycoprotein (NGAL), in a total of 20 review arms (9 that analyzed serum BMs, and 12 that analyzed BMs in urine). The population evaluated in the primary studies was predominantly adults. Two SRs included strictly adults, 5 reviews also included studies in the paediatric population, and 4 did not report the age groups. The results of the evaluation with the ROBIS tool showed that most of the reviews had a low overall risk of bias. CONCLUSIONS: There are 10 SRs of evidence relating to the diagnostic accuracy of serum and urinary biomarkers for lupus nephritis. Among the BMs evaluated, anti-C1q, urinary MCP-1, TWEAK, and NGAL stood out, highlighting the need for additional research, especially on LN diagnostic panels, and attempting to address methodological issues within diagnostic accuracy research. This would allow for a better understanding of their usefulness and possibly validate their clinical use in the future. REGISTRATION: This project is registered on the International Prospective Registry of Systematic Reviews (PROSPERO) database (CRD42020196693).

Anti-hyperglycemic, lipid-lowering, and anti-obesity effects of the triterpenes α and ß-amyrenones in vivo.

OBJECTIVE: Diabetes, obesity, and their associated metabolic disorders are public health problems that require prevention and new efficient drugs for treatment. We evaluated the anti-hyperglycemic, lipid-lowering, and anti-obesity effects of semisynthetic α, ß-amyrenones (ABA). MATERIALS AND METHODS: BALB/c mice were used for performing an acute model of oral carbohydrate and triglyceride tolerance, and in a streptozotocin-induced diabetes model, where glycemia and body weight changes were measured during ten days. C57BL/6 strain mice were used in the diet-induced obesity model, where lipidemia and body weight were measured during four weeks, and biochemical and histological parameters were analyzed after euthanasia. The doses considered in this study were 25, 50, and 100 mg/kg of ABA, used following some criteria for each experiment. RESULTS: ABA 25 mg/kg reduced the postprandial glycemia peak higher than acarbose 50 mg/kg (p<0.05). ABA 50 mg/kg significantly reduced glycemia in diabetic mice compared to acarbose 50 mg/kg (p<0.05). There was a reduction in the weight of the obese animals treated with ABA 25 and 50 mg/kg (p<0.05). ABA 50 mg/kg also significantly reduced lipidemia in these animals compared to orlistat 50 mg/kg. CONCLUSION: This study presents evidence of ABA's action in reducing postprandial glycemia and obesity in mice.

Eugenol and Lippia alba essential oils as effective anesthetics for the Amazonian freshwater stingray Potamotrygon wallacei (Chondrichthyes, Potamotrygonidae).

This study assessed the potential of eugenol and the essential oil of Lippia alba (EOLA) in providing suitable anesthetic induction and recovery times, and their consequent effects on the blood and respiratory physiology, as well as the gill architecture of an Amazonian freshwater stingray, Potamotrygon wallacei, shortly after reaching the recovery and 48 h later. Juveniles of P. wallacei were exposed to increasing concentrations of eugenol (75, 100, 125, and 150 µL L-1) and EOLA (150, 175, 200, and 225 µL L-1) in an immersion bath. Anesthetic induction was found to be faster with the use of eugenol compared to EOLA. On the other hand, the stingrays anesthetized with eugenol displayed a longer recovery time than those exposed to EOLA. The highest concentrations of eugenol caused moderate to severe histological changes in the gills. No significant changes were found for hematocrit and plasma energy metabolites in the stingrays anesthetized with all concentrations of both eugenol and EOLA shortly after reaching the recovery from the ansthesia, when compared to those recovered after 48 h. Investigations regarding the potential use of these natural anesthetics are unprecedented for freshwater stingray species, and 200 µL L-1 EOLA is recommended as the most suitable anesthetic for use in juveniles of P. wallacei.

Sex steroid hormones and the associated morphological changes in the reproductive tract of free-living males of the cururu stingray Potamotrygon wallacei.

This study analyzed sex steroid hormones and morphology of the reproductive tract in free-living adult males of the cururu stingray (Potamotrygon wallacei). These stingrays were captured during the transition between drought and rising waters in the middle Rio Negro basin, which coincides with the reproductive period (spermatogenesis and copulation) of this endemic species. Levels of the plasma steroid hormones testosterone (T), 17 ß-estradiol (E2), and progesterone (P4), the gonadosomatic and hepatosomatic indices, as well as histological characteristics of the testis and epididymis were evaluated. Three reproductive maturational stages were observed: regression males, capable to reproduce males, and active males. Plasma T levels were increasing and parallel with the rising water period in the middle Rio Negro basin and were associated with the progress of spermatogenesis. This was not observed for E2, which did not vary significantly among the different reproductive stages. The high P4 concentrations in males in regression and males capable of reproducing appeared to regulate the progression of spermatogenesis. Increases in this hormone in the plasma of active males was associated with spermiogenesis and spermiation events. These findings have not been previously observed in Amazonian freshwater stingrays and demonstrate that the reproductive activity of P. wallacei males is concomitant to the period of rising waters in the middle Rio Negro basin, and is regulated by the action of gonadal steroids, mainly testosterone.

Non-significant association between - 330 T/G polymorphism in interleukin-2 gene and chronic periodontitis: findings from a meta-analysis.

BACKGROUND: Chronic periodontitis (CP) is an immune-inflammatory disease that promotes tissue damage around the teeth. Among the several inflammatory mediators that orchestrate the periodontitis, there is the interleukin (IL)-2. Genetic variations in IL2 gene may be associated with the risk and severity of the disease. Contrary results are available in the literature with inconclusive findings and none meta-analysis to gather these data. METHODS: A literature search was performed for studies published before June 11, 2019 in diverse scientific and educational databases. The data was extracted by two investigators and the statistical evaluation was performed by Review Manager statistical program with heterogeneity (I2) and Odds Ratio (OR) with 95% of Confidence Intervals (CI) calculations and a sensitive analysis to assess the accuracy of the obtained results. The publication bias was evaluated by Begg' and Egger's test with Comprehensive meta-analysis software. The value of P < 0.05 was considered as significant. RESULTS: Five studies were identified in diverse ethnical groups with 1425 participants. The - 330 T/G polymorphism in IL2 gene was not significantly associated with CP in allelic evaluation (P > 0.05) as well as in the genotypic comparisons (P = 0.15). The Begg's test and the linear regression Egger's test did not show any evidence of publication bias risk (P > 0.05) which was corroborated by the absence of obvious asymmetry in Funnel plot graphic. CONCLUSIONS: This meta-analysis showed a non-significant association between - 330 T/G polymorphism in IL2 gene and CP in any allelic evaluation.

Determinantes sociais da saúde e prevenção secundária do câncer do colo do útero no Estado do Amazonas, Brasil / Social determinants of health and secondary prevention of cervical cancer in the State of Amazonas, Brazil

Modelo do estudo: este foi um estudo ecológico transversal e descritivo. Objetivo do estudo: conhecer os determinantes sociais de saúde associados com a prevenção secundária do câncer do colo do útero no período de 2010 a 2014. Metodologia: a unidade amostral foi os 62 municípios do estado do Amazonas. Os dados secundários utilizados foram do Sistema de Informação do Câncer do Colo do Útero (SISCOLO), do Sistema de Informação da Atenção Básica (SIAB), do Instituto Brasileiro de Geografia e Estatística (IBGE) e do Atlas de Desenvolvimento Humano do Brasil. Resultados: baixa cobertura do exame Preventivo do câncer do colo do útero para mulheres de 25 a 64 anos, sendo a faixa etária com menor adesão ao rastreio, a de 60 a 64 anos. A média de analfabetismo feminino na idade de 25 anos ou mais, no Estado, foi de 25,01%, e este indicador apresentou correlação com a baixa cobertura do exame Preventivo para o câncer do colo do útero em todos os anos. A pouca cobertura da Estratégia Saúde da Família também se associou a baixa adesão ao rastreio. Além disso, a Proporção de Municípios com Amostras Insatisfatórias dos exames citopatológicos apresentou-se acima da meta preconizada pelo Ministério da Saúde. Conclusão: estratégias de promoção da saúde voltadas a adesão ao rastreio devem considerar o nível de escolaridade das mulheres e o fortalecimento da Atenção Primária à Saúde pautada no rastreio não oportunístico; estas devem ser uma das prioridades das políticas públicas nos municípios do Estado.(AU)

Study design: This was a cross-sectional and ecological descriptive study. Study objective: Knowing the social determinants of health associated with the secondary prevention of cervical cancer from 2010 to 2014. Methods: The sample included the 62 counties of the state of Amazonas, and secondary data were used from the Cervical Cancer Information System, the Primary Care Information System, the Brazilian Institute of Geography and Statistics and the Atlas of Human Development of Brazil. Results: Low coverage of the preventive exam for cervical cancer for women aged 25 to 64 years was found. Also, the age group with the lowest adherence to screening was 60 to 64 years old. The average illiteracy rate among women aged 25 years and over, in the state, was 25.01%, and this indicator correlated with the low coverage of the preventive exam for cervical cancer every year. The low coverage of the Family Health Strategy was also as-sociated with low adherence to screening. The proportion of counties with unsatisfactory samples from the cytopathological examinations was above the goal recommended by the Ministry of Health. Conclusion: Health promotion strategies regarding adherence to screening should consider women's level of education, and the reinforcement of Primary Health Care based on non-opportunistic screening should be one of the priorities of public policies in the counties of the state. (AU)

Antileishmanial activity of extracts from Libidibia ferrea: development of in vitro and in vivo tests / Atividade anti-leishmania de extratos de Libidibia ferrea: desenvolvimento de testes in vitro e in vivo

ABSTRACT Treatment of cutaneous leishmaniasis (CL) is difficult due to the scarce number of drugs able to eliminate completely the intracellular form of the parasite. In the present study, the aim was to evaluate: i) phytochemical properties of extracts from Libidibia ferrea; ii) antileishmanial activity of extracts from L. ferrea against promastigotes and amastigotes of Leishmania (Leishmania) amazonensis and Leishmania (Viannia) guyanensis; iii) the effects of topical treatment using hydrogel containing active extract of L. ferrea on golden hamsters infected with L. (L.) amazonensis. Extracts from leaves, branches and fruits of L. ferrea were obtained with hexane and methanol and were tested by in vitro assays in promastigotes and murine macrophages J774 experimentally infected with amastigotes of Leishmania. Groups of hamsters with CL received topical treatment with a formulation of extract (10%) hydrogels, 50 mg.day-1 for 40 days. In vitro activity of FrMeOH (methanolic extract from fruits without seeds) resulted in significant reduction of viable promastigotes of L. (L.) amazonensis (IC50 of 15.4 µg.mL-1) and demonstrated inhibition potential of amastigote forms of L. (L.) amazonensis and L. (V.) guyanensis and low cytotoxicity in macrophages. The overall data of topical treatment with extract hydrogels (GelFrMeOH) showed that lesion sizes were significantly reduced (42.78%), with low parasite burden by RT-qPCR and culture analysis by microscopy examination, and with histopathological findings such as lower inflammatory cell infiltration 40 days after treatment. Chemical analysis demonstrated FrMeOH contains high levels of phenolic compounds. The results indicate a possible alternative therapy for CL using phytotherapics.

RESUMO O tratamento da leishmaniose cutânea (LC) é de difícil resultado, devido ao escasso número de fármacos capazes de eliminar completamente a forma intracelular do parasita. No presente estudo, objetivou-se avaliar: i) propriedades fitoquímicas dos extratos de Libidibia ferrea; ii) a atividade antileishmania de extratos de L. ferrea contra promastigotas e amastigotas de Leishmania (Leishmania) amazonensis e Leishmania (Viannia) guyanensis; iii) os efeitos do tratamento tópico utilizando hidrogel contendo extrato ativo de L. ferrea em hamsters dourados (Mesocricetus auratus) infectados com L. (L.) amazonensis. Extratos de folhas, galhos e frutos de L. ferrea foram obtidos com hexano e metanol e foram testados por ensaios in vitro contra promastigotas e macrófagos J774 infectados com amastigotas de Leishmania. Grupos de hamsters infectados receberam hidrogel tópico com extrato (10%), 50 mg.dia-1 durante 40 dias. A atividade in vitro de FrMeOH (extrato metanólico dos frutos) mostrou redução significativa de promastigotas de L. (L.) amazonensis (IC50 de 15,4 μg.mL-1), potencial de inibição de formas amastigotas de L. (L.) amazonensis e L. (V.) guyanensis (IC50 303,36 μg.mL-1) e baixa citotoxicidade em células de macrófagos J774. Os resultados destacaram que as lesões cutâneas de animais que receberam tratamento com hidrogel + extrato (GelFrMeOH) apresentaram redução significativa (42,78%), menos úlceração e redução da carga parasitária detectada por RT-qPCR, microscopia e análise em cultura e alterações histopatológicas, como menor infiltrado inflamatório após 40 dias de tratamento. A análise química demonstrou que FrMeOH contém altos níveis de compostos fenólicos. Os resultados apontam para uma possível terapia alternativa para CL utilizando fitoterápicos.

The mammary glands of the Amazonian manatee, Trichechus inunguis (Mammalia: Sirenia): morphological characteristics and microscopic anatomy.

The mammaries from carcasses of two female Amazonian manatees were examined. Trichechus inunguis possesses two axillary mammaries beneath the pectoral fins, one on each side of the body. Each papilla mammae has a small hole on its apex--the ostium papillare. The mammaries are covered by a stratified squamous keratinized epithelium. The epithelium of the mammary ducts became thinner more deeply in the tissue and varied from stratified to simple cuboidal. There was no evidence of glandular activity or secretion into the ducts of the mammary glands.

Attenuation of adjuvant-induced arthritis in rats by phonophoresis with an aqueous gel of the Amazonian plant Elaeoluma nuda (Sapotaceae).

BACKGROUND: Various species of the genus Pouteria (Elaeoluma) are used by the native population of Brazil because of, among other factors, their anti-inflammatory properties. The anti-inflammatory properties of the extract of the Amazonian plant Elaeoluma nuda were recently identified in prospective pharmacological studies. OBJECTIVES: The objective of this study was to assess the anti-inflammatory effect of phonophoresis with aqueous gel extract of E. nuda in rat adjuvant-induced arthritis. METHODOLOGY: Arthritis was induced in Lewis rats with an adjuvant. Phonophoresis with E. nuda gel was then administered daily and the results compared with those obtained with phonophoresis of diclofenac diethylammonium gel and ultrasound therapy without phonophoresis. Arthritis in the different groups was evaluated by plethysmometry. Proinflammatory cytokines TNF-α and IL-1α were quantified by cytometric bead array (CBA). RESULTS: The effect of phonophoresis of aqueous gel with E. nuda extract on arthritis in rats' paws (a 33% reduction compared with the controls) was the same as that produced by phonophoresis with diclofenac diethylammonium. Ultrasound therapy without phonophoresis produced no significant effect on the 21st day of therapy. There was a significant reduction in TNF-α and IL-1α levels in the group treated with phonophoresis with E. nuda gel (p=0.0042; p=0.0003, respectively). CONCLUSION: Our results demonstrate the anti-inflammatory effect of phonophoresis with E. nuda gel on cytokines TNF-α, IL-1α and adjuvant-induced arthritis.

Macroscopic aspects of Saimiri sciureus dura mater / Aspectos macroscópicos da dura mater de Saimiri sciureus

Saimiri sciureus is a small New World primate (NHP) commonly called macaco-de-cheiro that inhabits the tropical forests of the Amazon basin. Anatomical features are not well studied in most primates, and the encephalic morphology and related structures are still quite unknown. Comparative anatomy of the meninges in South American primates is still scarce. Dura mater, arachnoid and pia mater are a group of stratified layers that surrounds and promotes protection to the medulla spinalis. This study aimed to shed light on the anatomy of dura mater in Samiri sciureus in order to contribute to the neuroscience in primates. We investigated three young females and two males of S. sciureus. Specimens were fixed through perfusion with a 10 percent formaldehyde aqueous solution. In S. sciureus encephalus few gyrus and circunvolutions, and a very delicate system consisting of eight sinus venosus was found between the dura mater layers. Based on our findings, we can conclude that the Saimiri sciureus dura mater is quite similar to other mammals, however we detected a new sinus venosus formation at the level of parietal bone, named sinus parietalis, what appears to be its first description.

Saimiri sciureus, com nome comum de macaco-de-cheiro, é um pequeno primata do Novo Mundo (PNM) que habita as florestas tropicais da Bacia Amazônica. Os detalhes anatômicos de primatas são pouco conhecidos e a anatomia comparada de animais selvagens da América do Sul é escassa, mais especificamente, sobre as meninges de PNM. Meninges pertencem a um sistema de membrana responsável por envolver e proteger o Sistema Nervoso Central; consiste em três membranas: dura mater, aracnoide e pia mater. Este estudo objetivou a elucidação do comportamento anatômico da dura mater de S. sciureus com o intuito de contribuir para a área de neurociências em primatas. Foram investigados três fêmeas e dois machos jovnes de S. sciureus. Os espécimes de meninges foram fixados por perfusão usando solução aquosa 10 por cento de formaldeído. O encéfalo de S. sciureus demonstrou um número baixo de giros e circunvoluções, e um sistema delicado de oito seios venosos foi verificado entre as camadas da dura mater. Baseados em nossos achados podemos concluir que a dura mater de S. sciureus é similar aos de outros mamíferos, no entanto, apresenta a formação de um novo seio venoso em nível ao osso parietal, sendo denominado de seio parietal. Isto parece ser a primeira descrição deste seio.

Reproductive anatomy of the female Amazonian manatee Trichechus inunguis Natterer, 1883 (Mammalia: Sirenia).

The Amazonian manatee (Trichechus inunguis) is uniparous and has a slow reproduction cycle due to a long gestation period and long interval between births. Even though protected by law, hunting remains one of the main causes hindering the natural population growth of this species in the wild. The histology and reproductive anatomy provide information on the history and reproductive status of the female and offer a tool for the conservation of the species. The present study describes the anatomy of the female reproductive tract in T. inunguis. It is based on materials from three reproductive tracts fixed in 10% buffered formalin. The ovaries, uterine tubes, uterus, vagina, and external genitalia are described. The hymen presents two tiny openings separated by a segment that, upon rupturing during the first copulation, should make up a single vaginal opening. A still intact hymen and the absence of placental scars in the uterus were found in one specimen. Additionally, the presence of a hemorrhagic body and Graafian follicles on the right ovary were observed, as well as whitish scars and among them, possible corpora albicantia. These findings suggest that T. inunguis undergoes infertile estrus cycles before its first gestation. Macroscopically, counting of the whitish scars is hindered by the small diameter of these structures. It is not possible to differentiate between the scars resulting from ruptured (corpora albicantia) and nonruptured follicles (regressed corpora atretica). The presence of whitish scars on both ovaries of the same specimen suggests their bilateral function in T. inunguis.

Peripheral blood cells of the armored catfish Hoplosternum littorale Hancock, 1828: a morphological and cytochemical study

Cytochemical studies are used to identify fish leukocytes and as a basis for studying the functions of these cells in cellular immune responses. In this work, we investigated the morphological features and cytochemical properties of the blood cells in the armored catfish (Hoplosternum littorale), a South American teleost. Reticulocytes, which accounted for 8-24.6% of the red blood cell population, stained with brilliant cresyl blue and contained a granular material similar to residual RNA. Thrombocytes, lymphocytes, monocytes, neutrophils, heterophils and eosinophils were identified and characterized in blood smears stained with May Grünwald-Giemsa-Wright. The lymphocytes were small, round cells with a basophilic cytoplasm and contained no periodic acid-Schiff (PAS), peroxidase or non-specific esterase activity. The thrombocytes were usually fusiform, with a hyaline cytoplasm that was acidophilic when stained with alkaline toluidine blue. The monocytes were round, with a basophilic and sometimes vacuolated cytoplasm that contained non-specific esterase activity. The neutrophils were large and round, with typical neutrophilic granules that sometimes showed moderate staining. The nuclei were rod-shaped and occasionally segmented, with PAS-positive granules that gave a weak reaction for peroxidase. The heterophils were large and round with coarse eosinophilic and basophilic, PAS-positive granules. The eosinophils were round and medium-sized, with eosinophilic granules that generally gave a negative reaction in all cytochemical stainings. The marked variation in the granulocyte morphology of H. littorale meant that a standard analysis based only on the morphology of these cells was insufficient for identifying all of the cell types.