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Introduction:Testicular histology constitutes one of the least explored aspects in frogs of the genus Atelopus. This taxonomic group shows an alarming population decline; therefore, its reproductive biology is one of the greatest topics of interest for its conservation. Objective:To describe the testicular morphology and the spermatogenetic lineage cells in adult males of Atelopus laetissimus, Atelopus nahumae, andAtelopus carrikeriin the Sierra Nevada de Santa Marta, Colombia. Methods:During June – July 2017 and 2018, sampling was conducted in the localities of San Lorenzo and Páramo Cebolletas, Sierra Nevada de Santa Marta (SNSM), to collect 15 adult males, 5 per species. Testes samples were fixed in Bouin to be processed by the standard paraffin-embedding technique. Histological sections (3 mμ) were stained with Hematoxylin-eosin andMallory-Heidenhain-Azan-Gomori's. For the description and photographicregister of the germ cells, the photonic microscopy technique was used with the differential interference contrast system. Results:The testes are oval organs, compact, light yellow color, and with little vascularization. Externally, they are surrounded by a thin albuginea tunic constituted by regular dense connective tissue.Inside this layer, they are composed of numerous seminiferous tubules of hexagonal contour, in which germ cell cystsare distinguished at different stages of spermatogenesis (spermatogonia I and II, spermatocyte I and II, and early and late spermatids) and spermiogenesis (spermatozoa in fascicles and free spermatozoa). Separating the seminiferous structures is the interstitial tissue in which Leydig cells and blood vessels stand out.Additionally, in the cranial part of the testis, the Bidder's organ was found, formed by two distinguishableregions, the cortex and the medulla. In the cortex, there are previtellogénic oocytes of different sizes surrounded by a monolayer of flat follicular cells. For its part, the medullary region is the connective tissue that nourishes the oocytes and is constituted by blood capillaries. Conclusions:The gonads of the three species analyzed present a cystic cellular organization similar to other anurans, where all stages of spermatogenesis and spermiogenesis were identified, possibly indicating a continuous reproductive activity. Likewise, the Bidder’s organ is reported for the first time in the three Atelopus species, which allows suggesting a possible sexual reversion in case of a population decrease of females as a reproductive strategy. Introducción: La histología testicular constituye uno de los aspectos menos explorados en las ranas del género Atelopus. Este grupo taxonómico ostenta un declive poblacional alarmarte, es por ello, que su biología reproductiva resulta uno de los temas de mayor interés para su conservación. Objetivo:Describir la morfología testicular y las células del linaje espermatogénico en machos adultos de Atelopus laetissimus, Atelopus nahumae yAtelopus carrikerien la Sierra Nevada de Santa Marta, Colombia. Métodos:Durante Junio - Julio de 2017 y 2018 se realizaron muestreos en las localidades de San Lorenzo y Páramo Cebolletas, Sierra Nevada de Santa Marta (SNSM), para recolectar 15 machos adultos, 5 por especie. Las muestras de testículo se fijaron en Bouin para ser procesadas mediante la técnica estándar de inclusión en parafina. Las secciones histológicas (3 mμ) se tiñeron con Hematoxilina-eosina y Mallory-Heidenhain-Azan-Gomori's. Para la descripción y registro fotográfico de las células germinales, se utilizó la técnica de microscopía fotónica con el sistema de contraste diferencial de interferencia. Resultados:Los testículos son órganos ovalados, compactos, de color amarillo claro y con poca vascularización. Externamente, están rodeados por una delgada túnica albugínea constituida por tejido conectivo denso regular. Al interior de esta capa hay numerosos túbulos seminíferos de contorno hexagonal, en los que se distinguen quistes de células germinativas en diferentes etapas de la espermatogénesis (espermatogonia I y II, espermatocito I y II y espermátidas tempranas y tardías) y espermiogénesis (espermatozoides en fascículos y espermatozoides libres). Separando las estructuras seminíferas se halla el tejido intersticial en el que se destacan las células de Leydig y los vasos sanguíneos. Adicionalmente, en la parte craneal del testículo se encontró el órgano de bidder formado por dos regiones diferenciables, la corteza y la medula. En la corteza se aprecian ovocitos previtelogénicos en diferente tamaño rodeados por una monocapa de células foliculares planas. Por su parte, la región medular es el tejido conectivo que nutre los ovocitos y está constituido por capilares sanguíneos. Conclusiones:Las gónadas de las tres especies analizadas presentan una organización celular quística de manera similar con otros anuros, donde se identificó todos los estadios de la espermatogénesis y espermiogénesis indicando posiblemente una actividad reproductiva continua. Así mismo, se reporta por primera vez el órgano de bidder en las tres especies de Atelopus, lo cual permite sugerir una posible reversión sexual en caso de una disminución poblacional de las hembras como una estrategia reproductiva.

This study aimed to evaluate the effects of different temperatures on incubation variables, performance, and morphometry of the duodenal mucosa of Japanese quails (Coturnix coturnix japonica) submitted to chronic heat stress after hatching. We distributed 540 eggs in three incubators with a temperature of 37.8°C and 60% of humidity. From the 6th day of incubation until hatching, the temperatures were adjusted to (37.8°C, 38.5°C and 39.5°C). After hatching, quails were evaluated for the quality score, weighed, and distributed in a completely randomized design with three incubation temperatures (37.8, 38.5, and 39.5°C) and two ambient temperatures (stress and thermoneutral). At 10, 20, 30, and 40 days they were weighed to determine the live weight (g) and weight gain(g). To collect the duodenum and determine morphometric parameters, we euthanized four quails of each treatment. The data were analyzed, and the differences between the means determined by the Tukey test at 5%. The incubation temperature of 39.5°C provided lower hatching rate and the live weight at birth; however, from the 10th day of age, increased live weight, weight gain, and positively influenced the morphological parameters of the duodenal mucosa in situations of chronic stress. RESUMO Objetivou-se avaliar os efeitos de diferentes temperaturas de incubação sobre as variáveis de incubação, desempenho e morfometria da mucosa duodenal de codornas japonesas (Coturnix coturnix japonica) submetidas ao estresse térmico crônico por calor após eclosão. Foram distribuídos 540 ovos em três incubadoras, com temperatura de 37,8°C e umidade 60%. A partir do sexto dia de incubação até a eclosão, as temperaturas foram ajustadas para 37,8°C, 38,5°C e 39,5°C. Após a eclosão, as codornas foram avaliadas quanto ao escore de qualidade, pesadas e distribuídas em um delineamento inteiramente ao acaso, com três temperaturas de incubação (37,8ºC, 38,5ºC e 39,5°C) e duas temperaturas ambientes (estresse e termoneutro). Aos 10, 20, 30 e 40 dias, foram pesadas para determinar o peso vivo (g) e o ganho de peso(g). Quatro codornas de cada tratamento foram eutanasiadas para coleta do duodeno, para determinar os parâmetros morfométricos. Os dados foram analisados e as diferenças entre as médias foram determinadas pelo teste de Tukey a 5%. A temperatura de incubação de 39,5°C proporcionou menor taxa de eclosão e menor peso vivo ao nascer, entretanto, a partir do 10° dia de idade, essa temperatura aumentou o peso vivo, o ganho de peso e influenciou positivamente os parâmetros morfológicos da mucosa duodenal em situações de estresse crônico.

The objective of this study was to conduct a survey of the analysis of the supply profile of the discipline of histology in Biological Sciences courses in public higher education institutions in the state of Paraíba. This is a cross-sectional, descriptive and quantitative study, carried out in the year 2022, based on data available on the websites of public higher education institutions in the state of Paraíba. Thus, for the analysis it was verified which institutions offer the discipline of Histology in the graduation course of Biological Sciences; for this, the curricular matrix of each course was observed. According to the research findings, there is a total of 17 Biological Science courses offered in the state of Paraíba, where all offer the discipline of histology with a workload variation between 30h and 67h. Thus, after the analyses, it can be concluded that most public higher education institutions in the state of Paraíba offer the discipline of histology in their curricula. El objetivo de este estudio fue realizar una encuesta de análisis del perfil de la oferta de la disciplina de histología en los cursos de Ciencias Biológicas en las instituciones públicas de educación superior en el estado de Paraíba. Se trata de un estudio transversal, descriptivo y cuantitativo, realizado en el año 2022, a partir de los datos disponibles en los sitios web de las instituciones públicas de enseñanza superior del estado de Paraíba. Así, para el análisis se verificó qué instituciones ofrecen la disciplina de Histología en el curso de pregrado de Ciencias Biológicas; para ello, se observó la matriz curricular de cada curso. De acuerdo con los resultados de la investigación, hay un total de 17 cursos de Ciencias Biológicas ofrecidos en el estado de Paraíba, donde todos ofrecen la asignatura de histología con una variación de la carga de trabajo entre 30h y 67h. Así, tras el análisis, se puede concluir que la mayoría de las instituciones públicas de enseñanza superior del estado de Paraíba ofrecen la asignatura de histología en sus planes de estudio. O objetivo deste trabalho foi realizar um levantamento da análise do perfil de oferta da disciplina de histologia nos cursos de Ciências Biológicas nas instituições de ensino superior públicas do estado da Paraíba. Trata-se de um estudo transversal, descritivo e quantitativo, realizado no ano de 2022, a partir dos dados disponíveis nos sites das instituições de ensino superior públicas do estado da Paraíba. Dessa forma, para a análise foi verificado quais as instituições oferecem a disciplina de Histologia no curso de graduação de Ciências Biológicas, para isto, foi observado a matriz curricular de cada curso. Conforme os achados da pesquisa, existe um total de 17 cursos de Ciências Biológicas ofertados no estado da Paraíba, onde todos oferecem a disciplina de histologia com variação de carga horária entre 30h e 67h. Dessa forma, após as análises, pode-se concluir que a maior parte das instituições de ensino superior públicas do estado da Paraíba oferece a disciplina de histologia na sua grade curricular.

Los ácaros suelen ser ectoparásitos comunes de los vertebrados. Su distribución cor poral puede variar entre especies y pueden ocasionar lesiones en la piel. Los ácaros de Liolaemus pacha pertenecen al género Neopterygosoma y se ubican por debajo de las escamas de su hospedador. Los escasos registros sobre la relación ectoparásito- hospedador nos llevó a proponernos como objetivos, definir cuantitativamente la distribución corporal y determinar los posibles daños ocasionados por los ácaros en los sitios de inserción mediante estudios histopatológicos. Para el conteo de ácaros se definieron cinco regiones corporales y se realizaron cortes histológicos de las zonas de la piel que presentaban ácaros. En total, se analizaron 52 ejemplares de ambos sexos. Se observó una mayor concentración de ácaros en la región ventral, específicamente en los laterales del vientre y en la región gular. El análisis histológico mostró que no hay daños epidérmicos ni reacciones inflamatorias en la piel del hospedador.Por lo tanto, concluimos que los ácaros de L. pacha no producen daños locales en la piel, e incluso, su forma característica de inserción, pone en duda el tipo de alimentación hematófaga, reportada en otros estudios. Los resultados obtenidos aportan nuev información sobre la biología de Liolaemus pacha y Neoptorygosoma sp., permitiendo mejorar la interpretación de su interacción.

Philodryas nattereriis a snake known popularly as field-runner or board, founded in regions of caatinga and in the Brazilian semiarid region. These snakes have a well-developed Duvernoy gland, connected to a grooved tooth, specialized for toxin inoculation. The work aimed to perform the histological characterization of the Duvernoy gland of the P. nattererisnake. For the study, four pairs of animal glands from research projects in natural history and distribution of herpetofauna were used, which were registered and deposited in the NUROF Herpetological Collection. After dissection, the glands underwent histologicalprocessing, where they were fixed in 10% buffered formaldehyde, dehydrated in increasing concentrations of alcohol and later diaphanized in xylol. After inclusion in paraffin, the blocks were microtomized, with 5 μm cuts to make the slides. Hematoxylin-eosin and toluidine blue were used for staining. For the histochemical study, Schiff's periodic acid (PAS) and PAS + salivary amylase were used. Macroscopically, the Duvernoy gland had an ovulated, lobulated and elongated shape, with a whitish color. The average length was 6 mm for the left gland and 7 mm for the right. Regarding height measurement, the left and right glands had an average of 3 mm each. Photomicrographs revealed that the Duvernoy gland is lined by a dense connective tissue capsule, divided into lobes by septic connective tissue. Each lobe contains secretory tubules formed by serous cells, which indicates production of proteins as enzymes. Glands with mucous cells were observed in the excretory duct, which may be related to the production of glycoproteins. We conclude that the Duvernoy gland of the snake P.nattererifalls into the second histological category according to Taub (1967), which consists of a predominantly serous gland, with intercalated mucous cells Philodryas nattererié uma serpente conhecida popularmente como corre-campo ou tabuleiro, sendo encontrada em regiões de caatinga e no semiárido brasileiro. Essas serpentes possuem uma glândula de Duvernoy bem desenvolvida, conectada a um dente sulcado, especializado para a inoculação de toxinas. O trabalho teve como objetivo realizar a caracterização histológica da glândula de Duvernoy da serpente P. nattereri. Para o estudo foram utilizados quatro pares de glândulasde animais oriundos de projetos de pesquisa em história natural e distribuição da herpetofauna,tombadas e depositadas na Coleção Herpetológica do NUROF. Após a dissecção, as glândulas passaram pelo processamento histológico, onde foram fixadas em formol tamponado a 10%, desidratadas em concentrações crescentes de álcool e posteriormente diafanizadas em xilol. Após a inclusão em parafina, realizou-se a microtomia dos blocos, com cortes de 5 μm para a confecção das lâminas. Utilizou-se para a coloração hematoxilina-eosina e azul de toluidina. Para o estudo histoquímico, utilizou-se ácido periódico de Schiff (PAS) e PAS + Amilase salivar. Macroscopicamente, a glândula de Duvernoy possuía forma ovulada, lobulada e alongada, apresentando coloração esbranquiçada. A média do comprimento foi de 6 mm para a glândula esquerdae 7 mm para a direita. Com relação a medição da altura, as glândulas esquerda e direita tiveram uma média de 3 mm cada. As fotomicrografias revelaram que a glândula de Duvernoy é revestida por uma cápsula de tecido conjuntivo denso, dividida em lóbulos por septos de tecido conjuntivo. Cada lóbulo contém túbulos secretores formados por células serosas, o que indica produção de proteínas como enzimas. Foi observado glândulas com células mucosas no ducto excretor, o que pode estar relacionado à produção de glicoproteínas. Concluímos que a glândula de Duvernoy da serpente P.nattererienquadra-se na segunda categoria histológica segundo Taub (1967), que consiste em uma glândula predominantemente serosa, com células mucosas intercaladasprodução de proteínas como enzimas. Foi observado glândulas com células mucosas no ducto excretor, o que pode estar relacionado à produção de glicoproteínas. Concluímos que a glândula de Duvernoy da serpente P.nattererienquadra-se na segunda categoria histológica segundo Taub (1967), que consiste em uma glândula predominantemente serosa, com células mucosas intercaladas

In vitro culture of twin-scaling explants of Galanthus transcaucasicus with different concentrations of plant growth regulators (PGRs) including 0.5, 1, 2, 3, 4, 6, 8, and 10 mg L-1 naphthaleneacetic acid (NAA) and 0.5, 1, 2, 3, and 4 mg L-1 benzyladenine (BA) was studied. After 18 weeks, the number of regenerated bulblets and intensity of callus was measured. Subsequently, bulblets were transferred to a medium with 0.5, 1, 2, 3, and 4 mg L-1 NAA and 0.5, 1, 2, 3, and 4 mg L-1 BA and, after 15 weeks, the bulblets length and diameter were measured. The highest intensity of callus was obtained on 4 mg L-1 NAA or 8 mg L-1 NAA with 1 mg L-1 BA. The highest number of regenerated bulblets was detected with 6 mg L-1 NAA and 2 mg L-1 BA. The highest diameter of bulblets occurred on four mgL-1 NAA (9.4 mm), while the lowest was observed on 0.5 mg L-1 BA (1.83 mm). The analysis of genetic variation using ISSR revealed that there was no somaclonal variation among the regenerated plants from BA and low level of NAA, but there was a significant somaclonal variation at high concentrations of NAA. Resumo Objetivou-se estudar o cultivo in vitro de explantes de dupla escala de Galanthus transcaucasicus com diferentes concentrações de reguladores de crescimento: 0,5; 1; 2; 3; 4; 6; 8 e 10 mg L-1 de ácido naftalenoacético (ANA) e 0,5; 1; 2; 3 e 4 mg L-1 de benziladenina (BA). Após 18 semanas foram avaliados o número de bulbilhos regenerados e a porcentagem do calo. Posteriormente, os bulbilhos foram transferidos para um meio com 0,5; 1; 2; 3 e 4 mg L-1 de ANA e 0,5; 1; 2; 3 e 4 mg L-1 de BA e após 15 semanas foram medidos o comprimento dos bulbos e diâmetro. A maior intensidade de calo foi obtida com 4 mg L-1 de ANA ou 8 mg L-1 de ANA com 1 mg L-1 de BA. O maior número de bulbilhos regenerados foi obtido com 6 mg L-1 de ANA e 2 mg L-1 de BA. O maior diâmetro dos bulbilhos ocorreu com 4 mg L-1 ANA (9,4 mm), enquanto o menor foi observado com 0,5 mg L-1 BA (1,83 mm). A análise da variação genética usando ISSR não revelou variação somaclonal entre as plantas regeneradas a partir de BA e baixa concentração de ANA, mas houve variação somaclonal significativa em altas concentrações de ANA.

The Siluriformes, popularly known as catfishes, are probably the vertebrate group with the highest diversity of venomous animals, even though only approximately a hundred venomous catfishes are reported to date. Venomous catfishes might present a delivery system apparatus, formed by an unbranched ray at the leading edge of pectoral and dorsal fins (spine), which can be stiffened and pungent, while venom glands can be present at the surface of such spines and/or the axillary region. This work investigated the presence, morphology and distribution of glands and pectoral-fin delivery apparatus in the heptapterid Brachyrhamdia genus. Pectoral-fin spine external morphology was compared across all valid species in the genus, histological sections of the pectoral-fin spine and axillary regions of B. heteropleura and B. marthae were produced, and dissections of the pectoral girdle region of the mentioned species were analyzed. The histological sections confirmed the presence of pectoral-fin glands at the surface of the pectoral-fin spine of Brachyrhamdia species, and cellular morphology indicates these glands are probably venomous. Also, we found a piriform gland at the axillary region, whose cell morphology is like the reported for other catfishes. However, we cannot currently confirm or deny axillary gland participation in the venom delivery apparatus. This work constitutes the first report of venom glands in Brachyrhamdia, and the first description of Heptapteridae axillary glands.

Rastrelliger brachysomais a mariculture candidate species, but reproduction in captive fish has been problematic. This report examines the difference in the HPG axis, the neuroendocrine system and the development of reproductive tissues between captive vs. wild maleR. brachysoma. The gonadosomatic index (GSI) of sexually mature male wildR. brachysomawas 1.12±0.34 and 1.94±0.26 during the non-breeding and breeding seasons, respectively. CaptiveR. brachysomahad a GSI of 1.88±0.17. All wildR. brachysomawere in the late spermatogenic stage irrespective of seasons. Immunostaining results showed that sbGnRH-immunoreactive neurons were distributed in three areas of the brain, namely the nucleus periventricularis, nucleus preopticus and nucleus lateralis tuberis. Follicle stimulating hormone and luteinizing hormone immunoreactivities were also observed in the pituitary gland. The levels of brain sbGnRH and GtH mRNA were not significantly different between the non-breeding and breeding seasons, but captive fish displayed (times or percent difference) lower mRNA levels than wild fish. These results suggest that these hormones control the testicular development inR. brachysomaand that the impaired reproduction in captivity may be due to their relative lower expression levels of follicle stimulating hormone and luteinizing hormone genes., Rastrelliger brachysomaes una especie candidata para la piscicultura marina, pero la reproducción de individuos cautivos de esta especie ha sido problemática. Este estudio examina las diferencias en el eje hipotálamo-hipófisis-gónada (eje HPG), junto con el desarrollo de tejidos reproductivos, entre machos cautivos y salvajes deR. brachysoma. El índice gonadosomático (IGS) de machos salvajes deR. brachysomasexualmente maduros fue de 1.12±0.34 y 1.94±0.26 durante la estación no-reproductiva y reproductiva, respectivamente. LosR. brachysomacautivos mostraron un IGS de 1.88±0.17. Todos losR. brachysomasalvajes se encontraban en un estado de espermatogénesis tardío, independientemente de la estación. Los resultados de inmunotinción mostraron que las neuronas sbGnRH-inmunoreactivas se distribuían en tres áreas del cerebro, nucleus periventricularis, nucleus preopticus y nucleus lateralis tuberis. Se detectó inmunoreactividad para Fsh y Lh también en la hipófisis. Los niveles de mRNA de sbgnrh y gths en cerebro no fueron significativamente diferentes entre las estaciones de reproducción y no-reproducción, aunque se observaron niveles de mRNA menores (diferencia en nivel o porcentaje) en individuos cautivos que en salvajes. Estos resultados sugieren que las hormonas analizadas controlan el desarrollo testicular enR. brachysomay que la inhibición de la reproducción en cautividad podría ser debida a unos menores niveles de expresión relativa de los genes de Fsh y Lh.

After the growth-promoting antibiotics prohibition, intestinal health became an increasing concern worldwide in poultry farming. The intestinal histological evaluation is an inexpensive technique that brings relevant information, but in poultry, the immediate process of intestinal post-mortem autolysis interferes directly on the samples quality for histological analysis hindering a precise diagnosis. This study aimed to standardize a technique for broilers’ intestines sample collection and fixation for histological analysis. Seven broiler chickens received a standard diet until 23 days of age when they were euthanized. Fragments of duodenum, jejunum, and ileum were collected using three methods: intestine strips, transverse section, and Swiss roll and posteriorly fixed in 10% buffered formalin and bouin solution. Tissue samples were submitted for histological (number of villi and viable villi per field) and morphometrical (villi height, crypt depth and villi:crypt ratio) evaluations and the results analyzed statistically. A significant high number of villi and viable villi per field in all regions was observed in the Swiss roll method. In the duodenum (p= 0.0066) and jejunum (p= 0.0058) an interaction between the Swiss roll method and the fixative buffered formalin was observed in the viable and number of villi per field, respectively. Regarding the morphometrical analysis significant differences were observed, in the jejunum villi height sampling by the methods Swiss roll (1,157.66 ± 148.25 µm, p= 0.0015) that showed the highest mean. Deeper crypt depths were observed in the jejunum (156.59 ± 15.68 µm, p= 0.0002) and ileum (131.13 ± 15.01 µm, p= 0.0006) collect by the Swiss roll method. An interaction between the bouin fixative was also observed in the jejunum (p= 0.0223) for this variable. Duodenum sampling by transversal section (12.68 ± 1.45 µm, p= 0.0076) was the only segment that had a significant difference for villi:crypt ratio, showing the highest mean. It can be concluded that the Swiss roll technique was the best method for morphometrical evaluation of the chickens’ intestines, since the highest counts of villi per field and viable villi per field were obtained, while buffered formalin was considered as the best fixative. Após a proibição da utilização dos antimicrobianos promotores de crescimento, saúde intestinal se tornou uma preocupação crescente na avicultura mundialmente. A avaliação histológica intestinal é uma técnica de baixo custo que traz informações relevantes, porém nas aves, o imediato processo de autólise intestinal post-mortem interfere diretamente na qualidade da amostra para histologia dificultando um diagnóstico preciso. Este estudo objetivou a padronização de uma técnica para coleta e fixação do intestino de frangos de corte para análise histológica. Sete frangos de corte receberam uma dieta padrão até 23 dias de vida, quando foram eutanasiados. Fragmentos de duodeno, jejuno e íleo foram coletados utilizando três métodos: intestino aberto, corte transversal e rocambole, e posteriormente fixados em formalina tamponada 10% e solução de bouin. Amostras de tecidos foram submetidas a avaliações histológicas (número de vilosidades e vilosidades viáveis por campo) e morfométricas (altura de vilosidade, profundidade de cripta e relação vilosidade:cripta) e os resultados analisados estatisticamente. Um significativo maior número de vilosidades e vilosidades viáveis por campo foi observado, em todos os segmentos intestinais, no método rocambole. No duodeno (p= 0,0066) e jejuno (p= 0,0058) uma interação entre o método rocambole e o fixador formalina tamponada foi observado para as variáveis vilosidades viáveis e número de vilosidades por campo, respectivamente. Referente a análise morfométrica diferenças significativas foram observadas na altura de vilosidade no jejuno amostrado pelo método rocambole (1.157,66 ± 148,25 µm, p= 0,0015) que apresentou a maior média. Criptas mais profundas foram observadas no jejuno (156,59 ± 15,68 µm, p= 0,0002) e íleo (131,13 ± 15,01 µm, p= 0,0006) coletados pelo método rocambole. Uma interação entre o fixador bouin também foi observado no jejuno (p= 0,0223) para esta variável. Amostragem do duodeno pelo método corte transversal (12,68 ± 1,45 µm, p= 0,0076) foi o único segmento que teve uma diferença significativa para relação vilosidade:cripta apresentando a maior média. Pode-se concluir que o método rocambole foi o melhor para avaliação morfométrica do intestino de frangos de corte, uma vez que as maiores contagens de vilosidades e vilosidades viáveis por campo foram obtidas, enquanto formalina tamponada foi considerada o melhor fixador.

Origanum vulgare has been of great interest in academia and pharma industry due to its antioxidant, antifungal and antitumor properties. The present study aimed to find the anti-MRSA potential and in vivo toxicity assessments of O. vulgare. O. vulgare extract was used to monitor anti-MRSA activity in mice. Following MRSA established infection in mice (Mus musculus), treatment with O. vulgare was continued for 7 days. Autopsies were performed and re-isolation, gross lesion scoring and bacterial load in various organs were measured. Additionally, blood sample was analysed for hematological assays. Toxicity assessment of O. vulgare potential as medicine was done at 200 mg/kg and 400 mg/kg by evaluating liver and kidney functions. Bacterial load and gross lesion in lungs and heart were significantly low compared to positive control following O. vulgare treatment. Likewise, O. vulgare treated groups had hematological, neutrophil and TLC values similar to control groups. Increased AST, ALP and total bilirubin alongwith marked hepatocellular degeneration and distortion around the central vein, inflammatory cell infiltration, and cytoplasmic vacuolization of hepatic cells was observed at higher dose. It is concluded that crude extract of O. vulgare may contain beneficial secondary metabolites and in future may be explored for curing infectious diseases. Resumo Origanum vulgare tem despertado grande interesse na academia e na indústria farmacêutica devido às suas propriedades antioxidantes, antifúngicas e antitumorais. O presente estudo teve como objetivo encontrar o potencial anti-MRSA e avaliações de toxicidade in vivo de O. vulgare. O extrato de O. vulgare foi usado para monitorar a atividade anti-MRSA em camundongos. Após infecção estabelecida por MRSA em camundongos (Mus musculus), o tratamento com O. vulgare foi continuado por 7 dias. As autópsias foram realizadas e o reisolamento, pontuação das lesões grosseiras e carga bacteriana em vários órgãos foram medidos. Além disso, a amostra de sangue foi analisada para ensaios hematológicos. A avaliação da toxicidade do potencial de O. vulgare como medicamento foi feita com 200 mg / kg e 400 mg / kg, avaliando as funções hepática e renal. A carga bacteriana e as lesões graves nos pulmões e no coração foram significativamente baixas em comparação com o controle positivo após o tratamento com O. vulgare. Da mesma forma, os grupos tratados com O. vulgare apresentaram valores hematológicos, de neutrófilos e de TLC semelhantes aos grupos de controle. Aumento de AST, ALP e bilirrubina total juntamente com degeneração hepatocelular marcada e distorção ao redor da veia central, infiltração de células inflamatórias e vacuolização citoplasmática de células hepáticas foram observados em doses mais altas. Conclui-se que o extrato bruto de O. vulgare pode conter metabólitos secundários benéficos e, no futuro, pode ser explorado para a cura de doenças infecciosas.

The genus Brachyspira corresponds to the group of bacteria formerly classified into the genus Serpulina and includes several commensal and pathogenic intestinal spirochetes that affect pigs, poultry, and other animal species, including humans. In birds, some pathogenic species of this genus causes a condition known as avian intestinal spirochetosis, which remains underdiagnosed, thereby causing serious economic losses. Brachyspira is a fastidious organism that necessitates the employment of fast and efficient identification techniques. The aim of this study was to identify Brachyspira spp. using histology, immunohistochemistry (IHC) and fluorescence in situ hybridization (FISH) in formalin-fixed paraffin embedded (FFPE) tissue samples from the cecum of commercial poultry. Samples were collected from 129 birds aged between 35 and 45 days from commercial broiler farms. For evaluation, routine histology processing (H&E) and the histochemical technique, periodic acid–Schiff (PAS) were done. Additionally, FFPE tissue samples were evaluated for FISH and IHC. The histological lesions were analyzed and graded after H&E staining, and the goblet cells were counted and compared using PAS staining with the positive and negative samples obtained through FISH and IHC. For FISH, probes labeled with Brachyspira spp., B. pilosicoli, B. hyodysenteriae, and B. intermedia were used, whereas rabbit polyclonal antibody specific for Brachyspira spp. was used for IHC. Of 129 samples, 82 were positive with IHC and 86 were positive with FISH. The samples positive for the genus Brachyspira in the FISH technique were tested for B. pilosicoli, B. hyodysenteriae, and B. intermedia in which 56 were positive for B. pilosicoli, 75 for B. hyodysenteriae and 80 for B. intermedia. There was an increase in goblet cells in the samples positive for FISH and IHC. The techniques used were effective and gave corresponding results, thus serving as a fast and efficient tool for diagnosis. O gênero Brachyspira corresponde ao grupo de bactérias anteriormente classificadas no gênero Serpulina e inclui várias espiroquetas intestinais comensais e patogênicas que afetam suínos, aves e outras espécies animais, incluindo humanos. Em aves, algumas espécies patogênicas desse gênero causam uma condição conhecida como espiroquetose intestinal aviária, que permanece sub-diagnosticada, causando sérios prejuízos econômicos. Brachyspira é um organismo fastidioso que necessita do emprego de técnicas de identificação rápidas e eficientes. O objetivo deste estudo foi identificar Brachyspira spp. usando histologia, imunohistoquímica (IHQ) e hibridização fluorescente in situ (FISH) em amostras de tecido fixado em formalina e embebido em parafina (TFEP) do ceco de aves comerciais. As amostras foram coletadas de 129 aves com idades entre 35 e 45 dias em granjas comerciais. Para avaliação, o processamento histológico de rotina (H&E) e a técnica histoquímica, ácido periódico-Schiff (PAS) foram realizados. Além disso, as amostras de tecido TFEP foram avaliadas para FISH e IHC. As lesões histológicas foram analisadas e graduadas após coloração H&E, e as células caliciformes contadas e comparadas pela coloração PAS com as amostras positivas e negativas obtidas por FISH e IHC. Para FISH, foram utilizadas sondas marcadas com Brachyspira spp., B. pilosicoli, B. hyodysenteriae e B. intermedia, enquanto o anticorpo policlonal de coelho específico para Brachyspira spp. foi usado para IHC. De 129 amostras, 82 foram positivas com IHC e 86 foram positivas com FISH. As amostras positivas para o gênero Brachyspira pela técnica de FISH foram testadas para B. pilosicoli, B. hyodysenteriae e B. intermedia, sendo 56 positivas para B. pilosicoli, 75 para B. hyodysenteriae e 80 para B. intermedia. Houve aumento de células caliciformes nas amostras positivas para FISH e IHC. As técnicas utilizadas foram eficazes e deram os resultados correspondentes, servindo assim como uma ferramenta de diagnóstico rápida e eficiente.

Urban agriculture is very common in the cities, as Rio de Janeiro and adjacencies, being important as economic activity. However, this practice can create appropriate conditions for establishment and maintenance of life cycle of many parasites. Lymneid snails can act as intermediate hosts of many trematodes species, including those that infect humans and wild and domestic. In the present study, the snail Lymnaea columella was collected from an urban watercress plantations and the morphology of cercariae released by them was analyzed by light and scanning electron microscopy. The specimens were identified as xiphidiocercariae, belonging to the Cercariae Ornatae group, of the order Plagiorchiiformes. Histological examination of the digestive gland of the infected snails revealed the presence of developing sporocysts, with hemocitary infiltration and metaplasia in the epithelial tissue. Resumo Agricultura urbana é muito comum nas cidades, como Rio de Janeiro e adjacências, sendo uma importante atividade econômica de subsistência. No entanto, esta prática pode criar condições adequadas para o estabelecimento e manutenção do ciclo de vida de muitos parasitos. Os moluscos limneídeos podem atuar como hospedeiros intermédios de muitas espécies de trematódeos, incluindo aqueles que infectam humanos e animais selvagens e domésticos. No presente estudo, o molusco Lymnaea columella foi coletado de plantações urbanas de agrião e a morfologia das cercárias liberadas por eles foi analisada por microscopia eletrônica de varredura leve e de varredura. As larvas foram identificadas como xifidiocercárias, pertencentes ao grupo das Cercariae Ornatae, Ordem Plagiorchiiformes. Análises histológicas da glândula digestiva dos moluscos infectados evidenciaram esporocistos, com formação de granuloma e metaplasia do tecido epitelial.

CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior CNPq - Conselho Nacional de Desenvolvimento Científico e Tecnológico FAPEG - Fundação de Amparo à Pesquisa do Estado de Goiás Infecções do trato reprodutivo (TR) feminino podem acometer até 40% das vacas no pós-parto e são os maiores responsáveis pelo descarte involuntário, constituindo grandes prejuízos econômicos ao setor agropecuário. A resposta imune inata do TR é amplamente estudada com o intuito de compreender os mecanismos responsáveis pela produção do padrão de resposta inflamatória desencadeada pelas infecções uterinas. Objetivou-se verificar a resposta imune inata mediada IL-1β, IL-6 e IL-8 em diferentes regiões do TR de novilhas mestiças, frente ao desafio com lipopolissacarídeo (LPS) (n = 43) nas fases folicular (n = 10) e luteínica (n = 33) do ciclo estral, bem como, avaliar a dinâmica das mudanças celulares entre as estruturas anatômicas do TR com foco na porção cervical e suas transições, por meio das avaliações macroscópica e histológica (n = 3 em fase folicular e n = 3 emluteínica). Para tanto, foram colhidos fragmentos da porção cranial da vagina, transição vagina-cervix, porção cervical média, transição cervix-endométrio, carúncula uterina e endométrio intercaruncular. Observou-se resposta imune frente ao desafio com LPS em todas as porções do TR, independente da fase do ciclo estral, com maior produção de citocinas pelas transições cervicais, comparado à porção caruncular do endométrio. Além disso, a presença de células caliciformes na cervix e suas transições apresentam contribuição fundamental para a imunidade inata uterina. Concluiu-se produção interleucinas 1β, 6 e 8 apresenta comportamento diferente entre as regiões no trato reprodutivo de novilhas mestiças quando desafiadas com LPS e não há efeito do ciclo estral na indução desta reposta. Além disso, que a produção de muco pela cervix suas transições constituem a principal barreira uterina contra infecções microbianas devido a suas propriedades físico-químicas. Infections of the female reproductive tract (RT) may occur in up to 40% of the postpartum cows and are the major responsible for involuntary disposal, constituting a huge economic loss in the agricultural sector. The innate immune response of the RT is widely studied to comprehend the mechanisms responsible for the production of the inflammatory response pattern triggered by the uterine infections. We aimed to verify the innate immune response mediated by IL-1β, IL-6, and IL-8 in different regions of the reproductive tract of crossbred heifers challenged with lipopolysaccharide (LPS) (n=43) at the follicular (n=10) and luteal (n=33) phases of the estrous cycle, as well as to evaluate the cellular change dynamic between the anatomic structures of the RT, focusing in the cervical portion and its transitions, using macroscopic and histologic evaluations (n=3 in the follicular phase and n=3 in the luteal phase). For that we collected samples of the cranial portion of the vagina, vagina-cervix transition, cervix medial portion, cervix-endometrium transition, uterine caruncles, and intercaruncular endometrium. Besides that, the presence of caliciform cells at the cervix and its transitions present a fundamental contribution to the innate uterine immunity. We concluded that the production of interleukins 1β, 6, and 8 present different behavior between the regions of the reproductive tract of crossbred heifers when challenged with LPS and has no effect of the estrous cycle at the induction of this response. Moreover, the mucous production made by the cervix and its transitional portions constitute the main uterine barrier against microbial infections because of its chemical-physical properties. Tese (Doutorado) 2024-04-17

RESUMO Esse estudo objetivou avaliar sazonalmente a histomorfometria do epidídimo de Dermanura cinerea (Gervais, 1856) em um fragmento de Mata Atlântica no nordeste do Brasil. Foram utilizados 18 espécimes machos adultos capturados por redes de neblina. As coletas ocorreram mensalmente ao longo de dezoito meses, durante duas noites consecutivas e os dados meteorológicos foram fornecidos pelo Instituto Nacional de Meteorologia. Depois de eutanasiados, os espécimes tiveram os epidídimos coletados e esses órgãos foram fixados e processados. As lâminas histológicas foram coradas por Hematoxilina - Eosina e analisadas em microscopia óptica. Os parâmetros morfométricos analisados foram as áreas do túbulo, do lúmen e do epitélio das regiões do segmento inicial, cabeça, corpo e cauda do epidídimo. Os dados histomorfométricos obtidos foram submetidos às análises no teste U de Mann-Whitney. Os resultados revelaram que D. cinerea apresentou espermatozoides em todas as regiões do epidídimo, exceto no segmento inicial. As maiores médias das áreas tubular, do lúmen e do epitélio nas quatro regiões, foram constatadas durante os meses secos. Portanto, D cinerea apresentou maior sensibilidade na região da cauda do epidídimo, ao longo dos meses com baixos índices pluviométricos. Isso indica que as condições ambientais apresentam considerável influência sobre a morfofisiologia epidídimária dessa espécie, sobretudo, em relação ao armazenamento de espermatozoides na cauda desse órgão, em área de Mata atlântica do nordeste brasileiro. ABSTRACT Description and seasonal variation in epididymal histomorphometry of Dermanuracinerea (Chiroptera: Phyllostomidae) in a fragment of the Atlantic Forest in the Northeastern Brazil. This study aimed to evaluate seasonal patterns in the histomorphometry of the epididymis of Dermanura cinerea (Gervais, 1856) in a fragment of the Atlantic Forest in the Northeastern Brazil. Eighteen adult male specimens captured by mist net were used. The field-work occurred monthly over 18 months, during two consecutive nights. Meteorological data (precipitation) were obtained from the National Institute of Meteorology. After euthanasia, specimens had the epididymis collected, which were fixed and processed. The histological slides produced were stained by Hematoxylin - Eosin and analyzed by optical microscopy. The morphometric parameters analyzed were the tubular, lumen and epithelium areas, of the regions of the initial segment, caput, corpus and cauda of the epididymis. The histomorphometric data were submitted to the Mann-Whitney U test analyzes. The results showed that D. cinerea presented spermatozoa in all regions of the epididymis, except in the initial segment. The highest averages of the tubular, lumen and epithelial areas in the four regions were observed during the dry months. Therefore, D cinerea presented greater sensibility in the region of the cauda of the epididymis, during the months with low rainfall indices. This indicates that environmental conditions have considerable influence on the epididymal morphophysiology of this species, especially in relation to the storage of sperm in the tail of this organ, in area of Atlantic forest in northeastern Brazil.

Locker is a fungicide used in different crops for biological control, affecting both insect pests and pollinators, such as bees. This study aimed to evaluate toxicity and establish the lethal concentration (LC50) of the fungicide Locker in stingless bees Scaptotrigona bipunctata, investigating changes in brain cells chromatin structure and in midgut morphology. This agrochemical is a combined fungicide that contains three active ingredients: benzimidazole, triazole and strobilurin. Adult forage bees were orally exposed to the following concentrations: 1.7 mg a.i./L; 2.55 mg a.i./L; 3.4 mg a.i./L and 4.25 mg a.i./L of fungicide, and submitted to histochemical and morphological analysis 24, 48 and 72 hours after ingestion. The analysis of S. bipunctata brain cells showed changes in chromatin condensation in all treatments. There were also several morphological changes in the midgut at all concentrations and exposure times. Thus, the fungicide Locker significantly affected bee survival, inducing changes in their internal morphology, which can lead to alterations in their activities, directly interfering in their foraging behavior and survival. Locker es un fungicida utilizado en diferentes cultivos para el control biológico, que afecta tanto a insectos plagas como a polinizadores, como las abejas. Este estudio tuvo como objetivo evaluar la toxicidad y establecer la concentración letal (CL50) del fungicida Locker en abejas sin aguijón Scaptotrigona bipunctata, investigando cambios en la estructura de la cromatina de las células cerebrales y en la morfología del intestino de la abeja. El producto utilizado es un fungicida combinado que contiene tres principios activos: benzimidazol, triazol y estrobilurina. Las abejas forrajeras adultas fueron expuestas por vía oral a concentraciones de 1,7 mg a.i./L; 2,55 mg a.i./L; 3,4 mg a.i./L y 4,25 mg a.i./L de fungicida, y sometidos a análisis histoquímico y morfológico 24, 48 y 72 horas después de la ingestión del producto. El análisis de células cerebrales de S. bipunctata mostró cambios en la condensación de cromatina en todos los tratamientos. También hubo varios cambios morfológicos en el intestino medio en todas las concentraciones y tiempos de exposición. Así, el fungicida Locker afectó significativamente la supervivencia de las abejas, induciendo cambios en su morfología interna, lo que puede conducir a cambios en sus actividades, interfiriendo directamente en su comportamiento de forrajeo y supervivencia. Locker é um fungicida utilizado em diferentes culturas para controle biológico, afetando tanto insetos pragas quanto polinizadores, como as abelhas. Este estudo teve como objetivo avaliar a toxicidade e estabelecer a concentração letal (CL50) do fungicida Locker em abelhas sem ferrão Scaptotrigona bipunctata, investigando alterações na estrutura da cromatina das células cerebrais e na morfologia do intestino médio das abelhas. O produto utilizado é um fungicida combinado que contém três ingredientes ativos: benzimidazol, triazol e estrobilurina. Abelhas forrageiras adultas foram expostas via oral às concentrações de 1,7 mg i.a./L; 2,55 mg a.i./L; 3,4 mg i.a./L e 4,25 mg i.a./L de fungicida, e submetidas à análises histoquímicas e morfológicas após 24, 48 e 72 horas da ingestão do produto. A análise das células cerebrais de S. bipunctata mostrou alterações na condensação da cromatina em todos os tratamentos. Houve também várias mudanças morfológicas no intestino médio em todas as concentrações e tempos de exposição. Dessa forma, o fungicida Locker afetou significativamente a sobrevivência das abelhas, induzindo alterações em sua morfologia interna, que podem levar a alterações em suas atividades, interferindo diretamente em seu comportamento de forrageamento e sobrevivência.

Introduction: Helicobacter pylori (H. pylori) is a Gram negative bacterium considered to be the etiologic agent of various gastric diseases. The prevalence of bacterial infection varies according to age, geographic location, ethnicity and socioeconomic status. The chronic infection caused by microorganism can favor the development of severe pathologies such as gastric adenocarcinoma. In this sense, early diagnosis is essential for a better prognosis and therapeutic success. Several diagnostic methods performed using invasive and non-invasive techniques, with different sensitivity and specificity, have been used in the detection of H. pylori. Objective: To compare the performance of the molecular and histopathological technique used in the diagnosis of H. pylori infection. Methods: 76 gastric tissue samples were collected from dyspeptic patients who underwent molecular and histopathological diagnosis. Molecular detection was performed using the ribosomal gene (16S rRNA) using the polymerase chain reaction (PCR) technique. Results: The PCR-based molecular diagnostic method detected the bacterium in 63.1% of the samples, while the histopathological test identified the microorganism in only 38.1% of gastric biopsies. The data demonstrated that the PCR technique was about 1.6 times more sensitive than the histopathological technique. Conclusion: The PCR technique was the most efficient diagnostic method for detecting H. pylori and can be implemented in the laboratory routine as a complementary test for the early detection of H. pylori. RESUMEN Introducción: Helicobacter pylori (H. pylori) es una bacteria Gram negativa considerada agente etiológico de diversas enfermedades gástricas. La prevalencia de la infección bacteriana varía según la edad, la ubicación geográfica, la etnia y el nivel socioeconómico. La infección crónica provocada por esos microorganismos puede favorecer el desarrollo de patologías graves como el adenocarcinoma gástrico. Por esa razón, el diagnóstico precoz es fundamental para un mejor pronóstico y éxito terapéutico. En la detección de H. pylori se han utilizado varios métodos de diagnóstico realizados mediante técnicas invasivas y no invasivas, con diferentes sensibilidad y especificidad. Objetivo: Comparar el desempeño de las técnicas moleculares e histopatológicas utilizadas en el diagnóstico de la infección por H. pylori. Métodos: Se recolectaron 76 muestras de tejido gástrico de pacientes dispépticos y se las sometieron a diagnóstico molecular e histopatológico. La detección molecular se realizó mediante el gen ribosómico (ARNr 16S) mediante la técnica de reacción en cadena de la polimerasa (PCR). Resultados: El método molecular de diagnóstico basado en PCR detectó la bacteria en el 63,1% de las muestras, mientras que la prueba histopatológica identificó el microorganismo en solo el 38,1% de las biopsias gástricas. Los datos demostraron que la técnica de PCR era aproximadamente 1,6 veces más sensible que la técnica histopatológica. Conclusión: La técnica de PCR fue el método diagnóstico más eficaz para la detección de H. pylori y puede implementarse en la rutina del laboratorio como prueba complementaria para la detección precoz de H. pylori. RESUMO Introdução: Helicobacter pylori (H. pylori) é uma bactéria Gram negativa considerada o agente etiológico de várias doenças gástricas. A prevalência da infecção bacteriana varia de acordo com idade, localização geográfica, etnia e status socioeconômico. A infecção crônica ocasionada por esse microrganismo pode favorecer o desenvolvimento de patologias severas, como o adenocarcinoma gástrico. Nesse sentido, o diagnóstico precoce é essencial para um melhor prognóstico e o sucesso terapêutico. Vários métodos de diagnóstico realizados com técnicas invasivas e não invasivas, com diferentes sensibilidade e especificidade, têm sido utilizados na detecção de H. pylori. Objetivo: Comparar o desempenho das técnicas molecular e histopatológica utilizadas no diagnóstico da infecção por H. pylori. Métodos: Setenta e seis amostras de tecido gástrico foram coletadas de pacientes dispépticos e submetidas ao diagnóstico molecular e histopatológico. A detecção molecular foi realizada utilizando o gene ribossomal (rRNA 16S) por meio da técnica de reação em cadeia da polimerase (PCR). Resultados: O método molecular de diagnóstico com base na PCR detectou a bactéria em 63,1% das amostras, enquanto o teste histopatológico identificou o microrganismo em apenas 38,1% das biópsias gástricas. Os dados demonstram que a técnica de PCR apresentou cerca de 1,6 vezes mais sensibilidade que a técnica histopatológica. Conclusão: A técnica de PCR foi o método de diagnóstico mais eficiente para detecção de H. pylori e pode ser implementada na rotina laboratorial como teste complementar para detecção precoce de H. pylori.

Fil: Wang, Jue. University of Texas Health Science Center at Tyler. Department of Cellular and Molecular Biology; Estados Unidos Fil: Hertz, Laura. Saarland University. Theoretical Medicine and Biosciences; Alemania Fil: Hertz, Laura. Saarland University. Experimental Physics, Dynamics of Fluids; Alemania Fil: Ruppenthal, Sandra. Saarland University. Experimental Physics, Dynamics of Fluids; Alemania Fil: Ruppenthal, Sandra. Saarland University Hospital. Gynaecology, Obstetrics and Reproductive Medicine; Alemania Fil: El Nemer, Wassim. Aix Marseille Université. Etablissement Français du Sang PACA-Corse; Francia Fil: El Nemer, Wassim. Laboratoire d’Excellence GR-Ex; Francia Fil: Connes, Philippe. Laboratoire d’Excellence GR-Ex; Francia Fil: Connes, Philippe. University Claude Bernard Lyon 1. Vascular Biology and Red Blood Cell Teal. Laboratory LIBM EA7424; Francia Fil: Goede, Jeroen S. Kantonsspital Winterthur. Division of Oncology and Hematology; Suiza Fil: Bogdanova, Anna. University of Zürich. Institute of Veterinary Physiology. Red Blood Cell Research Group; Suiza Fil: Birnbaumer, Lutz. Pontificia Universidad Católica Argentina. Facultad de Ciencias Médicas. Instituto de Investigaciones Biomédicas; Argentina Fil: Birnbaumer, Lutz. National Institute of Environmental Health Sciences. Neurobiology Laboratory; Estados Unidos Fil: Kaestner, Lars. Saarland University. Theoretical Medicine and Biosciences; Alemania Fil: Kaestner, Lars. Saarland University. Experimental Physics, Dynamics of Fluids; Alemania Abstract: (1) Background: It is known that sickle cells contain a higher amount of Ca2+ compared to healthy red blood cells (RBCs). The increased Ca2+ is associated with the most severe symptom of sickle cell disease (SCD), the vaso-occlusive crisis (VOC). The Ca2+ entry pathway received the name of Psickle but its molecular identity remains only partly resolved. We aimed to map the involved Ca2+ signaling to provide putative pharmacological targets for treatment. (2) Methods: The main technique applied was Ca2+ imaging of RBCs from healthy donors, SCD patients and a number of transgenic mouse models in comparison to wild-type mice. Life-cell Ca2+ imaging was applied to monitor responses to pharmacological targeting of the elements of signaling cascades. Infection as a trigger of VOC was imitated by stimulation of RBCs with lysophosphatidic acid (LPA). These measurements were complemented with biochemical assays. (3) Results: Ca2+ entry into SCD RBCs in response to LPA stimulation exceeded that of healthy donors. LPA receptor 4 levels were increased in SCD RBCs. Their activation was followed by the activation of Gi protein, which in turn triggered opening of TRPC6 and CaV2.1 channels via a protein kinase C and a MAP kinase pathway, respectively. (4) Conclusions: We found a new Ca2+ signaling cascade that is increased in SCD patients and identified new pharmacological targets that might be promising in addressing the most severe symptom of SCD, the VOC.

Computational histomorphometry is an available and easy tool that has been used in the assessment of morphophysiological tissue changes, offering greater scientific reliability to the data, as well as facilitating the automation process. The present work aimed to describe the application of the methodology of the free software ImageJ for morphological evaluation of fish tissues. For this, micrographs of histological sections of the intestinal tract of fish stained with Periodic Acid-Schiff (PAS) were used as a model. The images were analyzed for variables of length, width, and tissue area and, number of cells or molecules. The application of computational histomorphometry demonstrated efficiency in the evaluation of histological structures of the intestine of fish supplemented with probiotics, contributing to the improvement of image analysis techniques in animal tissue models. La histomorfometría computacional es una herramienta disponible y fácil que se ha utilizado en la evaluación de cambios morfofisiológicos tisulares, ofreciendo mayor confiabilidad científica a los datos, además de facilitar el proceso de automatización. El presente trabajo tenido como objetivo describir la aplicación de la metodología de uso del software libre ImageJ en la evaluación morfológicos en tejidos de peces. Para ello, se utilizaron como modelo micrografías de cortes histológicos del tracto intestinal de peces teñidos con Ácido Periódico-Schiff (PAS). Las imágenes se analizaron en busca de variables de longitud, ancho y área de tejido y número de células o moléculas. La aplicación de histomorfometría computacional demostró eficiencia en la evaluación de estructuras histológicas del intestino de peces, contribuyendo a la mejora de las técnicas de análisis de imagen en modelos de tejidos animales. A histomorfometria computacional é uma ferramenta disponível e de fácil utilização, que tem sido utilizada na avaliação de alterações morfofisiológicas dos tecidos, oferecendo maior confiabilidade científica aos dados, além de facilitar o processo de automação. O presente trabalho teve como objetivo descrever a aplicação da metodologia do software livre ImageJ para avaliação morfológica de tecidos de peixes. Para isso, utilizou-se fotomicrografias de cortes histológicos do trato intestinal de peixes corados com Periodic Acid-Schiff (PAS) foram utilizadas como modelo. As imagens foram analisadas quanto às variáveis de comprimento, largura e área do tecido e número de células ou moléculas. A aplicação da histomorfometria computacional demonstrou eficiência na avaliação das estruturas histológicas do intestino de peixes, contribuindo para o aprimoramento das técnicas de análise de imagens em modelos de tecidos animais.

Objetivo: o objetivo desse trabalho foi de identificar se o desmame precoce é capaz de ocasionar alterações morfológicas no tecido gástrico de ratos wistar. Método: Foram dividimos os filhotes de ratos em dois grupos, Grupo Desmame (n=6) e Grupo Controle (n=6). O Grupo Desmame foi desmamado no 15°dia de vida por separação materna, enquanto o Grupo Controle permaneceu com suas mães amamentando. Com 90 dias, os grupos foram submetidos a eutanásia e em seguida a retirada do estômago para o processamento histológico. As lâminas foram fotomicrografadas e analisadas. Resultado: Os dados mostraram que o desmame precoce aumenta a área da mucosa, aumentando assim a área das glândulas e da muscular da mucosa, com isso aumenta também a acidez estomacal. Também observamos que o desmame diminuiu a área da muscular longitudinal e submucosa, fazendo com que o organismo sofra adaptações para o funcionamento. Mostrando assim os riscos que o desmame precoce pode ocasionar a saúde.

Background Cratera is a genus of land planarians endemic to the Brazilian Atlantic forest. The species of this genus are distinguished from each other by a series of external and internal characters, nonetheless they represent a challenging taxonomic issue due to the extreme alikeness of the species analysed in the present work. To resolve these difficulties, we have performed morphological analyses and used three nuclear markers (ribosomal 18S and 28S, Elongation Factor, a new anonymous marker named Tnuc813) and two mitochondrial fragments (Cytochrome c oxidase subunit I gene, and a fragment encompasing NADH deshydrogenase subunit 4 gene, trnF and the beginning of the Cytochrome c oxidase I gene) in an integrative taxonomic study. Methods To unveil cryptic species, we applied a molecular species delimitation approach based on molecular discovery methods, followed by a validation method. The putative species so delimited were then validated on the basis of diagnostic morphological features. Results We discovered and described four new species, namely Cratera assu, C. tui, C. boja, and C. imbiri. A fifth new species, C. paraitinga was not highly supported by molecular evidence, but was described because its morphological attributes are unique. Our study documents for the genus Cratera the presence of a number of highly similar species, a situation that is present also in other genera of land planarians. The high number of poorly differentiated and presumably recent speciation events might be explained by the recent geological history of the area.

Our objective was the characterization and staging of histological lesions in different anatomical sites of the central nervous system (CNS) of rabid cattle. The severity of the lesions was compared with the clinical stages of the disease, the variants of viral isolates, and with the load of virus. Thirty-one spontaneously affected rabid cattle the state of Santa Catarina underwent clinical follow-up and were eventually necropsied. CNS tissues were sampled and submitted to direct fluorescent antibody technique (DFAT), immunohistochemistry (IHC), routine histopathology with hematoxylin and eosin stain (HE), reverse transcriptase polymerase chain reaction (RT-PCR), and polymerase chain reaction in quantitative reverse transcriptase in real time (qRT-PCR). Affected cattle were allotted in four groups according to their clinical stage when euthanized: G1, euthanized while standing; G2, euthanized when in sternal recumbence; G3, euthanized when in lateral recumbence; and G4, affected cattle with natural death. In order to evaluate the degree of severity of the lesions and the presence of Negri bodies (NBs), the brain was sectioned at 9 sites. Additionally, spinal cord and trigeminal ganglion sections were examined. The intensity of the lesions was graded as either absent, mild, moderate, or marked, and the presence or absence of the NBs was noted. Histological lesions were characterized by lymphocytic and monocytic meningoencephalitis with NBs in 28 cases. In all analyzed groups, intensities of histological lesions ranging from mild to severe were observed. Brain regions with the highest inflammatory lesion intensity were the medulla at the level of obex, followed by the colliculus and thalamus. NBs were observed in a higher percentage in the cerebellum, followed by medulla at the obex level, striatum complex, and frontal telencephalon. The duration of the clinical course of the disease did not influence the intensity of the inflammatory lesion, but it did influence the presence of NBs, with a higher percentage of these inclusions in cattle that died naturally than in euthanized cattle. All isolated rhabdovirus included in this study were genetically compatible with samples from hematophagous bats Desmodus rotundus. The evaluation by qRT-PCR did not demonstrate a correlation between lesion intensity and the amount of virus. RESUMO: Nosso objetivo foi a caracterização e estadiamento de lesões histológicas em diferentes locais anatômicos do sistema nervoso central (SNC) de bovinos raivosos. A gravidade das lesões foi comparada com os estágios clínicos da doença, as variantes dos isolados virais e com a quantidade de vírus. Trinta e um bovinos do estado de Santa Catarina, afetados naturalmente por raiva, foram acompanhados clinicalmente e, ao final, necropsiados. Os tecidos do SNC foram amostrados e submetidos a imunofluorescência direta, imunohistoquímica, histopatologia de rotina, reação em cadeia da polimerase via transcriptase reversa (RT-PCR) e reação em cadeia da polimerase em transcriptase reversa quantitativa em tempo real (qRT-PCR). Os bovinos afetados foram distribuídos em quatro grupos, de acordo com sua fase clínica: G1, eutanasiados quando ainda se mantinham em pé; G2, eutanasiados quando em decúbito esternal; G3, eutanasiados quando em decúbito lateral; e G4, bovinos afetados com morte natural. Para avaliar o grau de gravidade das lesões e a presença de corpúsculos de Negri (CNs), o cérebro foi seccionado em 9 locais. Além disso, seções da medula espinhal e do gânglio trigêmeo foram examinadas. A intensidade das lesões foi graduada como ausente, leve, moderada ou acentuada, e a presença ou ausência dos CNs foi anotada. Lesões histológicas foram caracterizadas por meningoencefalite linfocítica e monocítica com CNs em 28 casos. Em todos os grupos analisados foram observadas intensidades de lesões histológicas variando de leve a grave. As regiões cerebrais com maior intensidade de lesão inflamatória foram o bulbo no nível do obex, seguido do colículo e tálamo. CNs foram mais prevalentes no cerebelo, seguido pelo bulbo ao nível do óbex, corpo estriado e telencéfalo frontal. A duração do curso clínico da raiva não influenciou a intensidade da lesão inflamatória, mas influenciou a presença de CNs, com maior porcentagem dessas inclusões em bovinos que morreram naturalmente do que em bovinos sacrificados. Todos os isolados rabdovírus obtidos neste estudo eram geneticamente compatíveis com amostras provenientes de morcegos hematófagos Desmodus rotundus.

This study aimed to identify gross and microscopic changes, caused by high-energy diets, in the rumen environment and hoof of confined beef cattle. The study sample comprised 40 confined heifers (Bos taurus) with no disease history divided into four experimental groups using different diets: Group 1 (D1, control), 48:52 forage:concentrate ratio; Group 2 (D2), 30:70 forage:concentrate ratio; Group 3 (D3), 30:70 forage:concentrate ratio + sucrose; Group 4 (D4), 100% concentrate. All animals underwent clinical examination, assessment of ruminal fluid pH and lameness, and sample collection after slaughter for histopathology of the hoof laminae and digital cushion and ruminal tissue. All dependent variables of the study were compared using the SPSS 20.0 statistical software. The variables that did not show normality (HR, RM, ST, and CRT) were compared with application of the Kruskal-Wallis test followed by the Dunnet’s multiple comparison test. All other variables were submitted to analysis of variance (ANOVA) followed by Tukey’s test. The different diets had an impact on the rumen environment (p

Fish feeding habit associated to the morphology of their digestive tube can provide useful biological and ecological information. Between January and December 2012, we examined 16 fish species discarded by trawl in the Potiguar basin, northeastern Brazil, and classified them into four groups according to Alimentary Index - IAi. Our aims were to: (1) provide information on diet composition of one species represented by each group (Opisthonema oglinum, Conodon nobilis, Larimus breviceps and Sphoeroides testudineus), (2) examine the morphology of their digestive tube, and (3) associate ingested food items with the morphological features. Stomach contents were collected and classified according to IAi: main, accessory and secondary categories; fragments of esophagus, stomach, and small intestine were examined using paraffin section technique. Biometric data (total length-TL, standard length-SL and total weight-TW), stomach weight-SW, and gonadal weight-GW were recorded and applied in two indices: (1) the stomach Repletion Index - SRI (SRI = SW/TW × 100) and (2) the gonadosomatic index - GSI (GSI = GW/TW × 100) for the analyses of feeding and reproductive activities, respectively. The main food categories were (1) Teleostei for C. nobilis, (2) Crustacea for L. breviceps and O. oglinum, and (3) Mollusca for S. testudineus. The feeding activity was higher during the dry season, while the GSI was higher in the rainy season. The morphological analysis revealed a regular structure of the tissues analyzed. We highlight the presence of tubular gastric glands and strong muscular walls in the stomach (O. oglinum; L. breviceps); well-developed pyloric caecum in the intestine (O. oglinum; C. nobilis; L. breviceps); and presence of pseudo-stomach (S. testudineus). Our study confirms that the digestive tube of the examined species has morphological adequacy for carnivorous feeding habit, providing useful information on tissue structures associated to diet composition. Resumo: O hábito alimentar de peixes associado à morfologia do tubo digestivo pode fornecer informações biológicas e ecológicas úteis. Entre janeiro e dezembro de 2012, examinamos 16 espécies de peixes descartadas por pesca de arrasto na Bacia Potiguar, nordeste do Brasil, e as classificamos em quatro grupos, de acordo com o Índice Alimentar-IAi. Nossos objetivos foram: (1) fornecer informações sobre a composição da dieta de uma espécie representante de cada grupo (Opisthonema oglinum, Conodon nobilis, Larimus breviceps e Sphoeroides testudineus), (2) examinar a morfologia de seu tubo digestivo e (3) associar os itens alimentares ingeridos com as características morfológicas descritas. O conteúdo estomacal foi coletado e classificado de acordo com o IAi nas categorias: principal, acessória e secundária; fragmentos de esôfago, estômago e intestino delgado foram examinados pela técnica de secção em parafina. Os dados biométricos (comprimento total-CT, comprimento padrão-CP e peso total-PT), peso do estômago-PE e peso gonadal-PG foram registrados e aplicados em dois índices: (1) Índice de Repleção Estomacal - IR (IR = PE / PT × 100) e (2) Índice Gonadossomático - IGS (IGS = PG / PT × 100) para análises de atividade alimentar e reprodutiva, respectivamente. As principais categorias alimentares foram: (1) Teleostei para C. nobilis, (2) Crustacea para L. breviceps e O. oglinum, e (3) Mollusca para S. testudineus. Maior atividade alimentar ocorreu durante a estação seca, e o IGS foi maior na estação chuvosa. O exame morfológico revelou estrutura regular dos tecidos analisados, destacando-se a presença de glândulas gástricas tubulares e camadas musculares espessas no estômago (O. oglinum; L. breviceps); cecos pilóricos bem desenvolvidos no intestino (O. oglinum; C. nobilis; L. breviceps); e presença de pseudoestômago (S. testudineus). Nosso estudo confirma que o tubo digestivo das espécies examinadas possui adequação morfológica ao hábito alimentar carnívoro e fornece informações úteis sobre as estruturas dos tecidos associados à composição da dieta.

Cardinal tetra Paracheirodon axelrodi and bloodfin tetra Aphyocharax anisitsi are two species of characids with high trade value as ornamental fish in South America. Although both species inhabit middle water layers, cardinal neon exhibits a tropical distribution and bloodfin tetra a subtropical one. Generally, these species are difficult to grow, so it becomes essential to know some key components of the neuroendocrine system to achieve their reproduction in captivity. Considering the importance of deepening the knowledge of the reproductive physiology through functional morphology, for the first time in this work we performed an anatomical, morphological and immunohistochemical analysis of the pituitary gland of these two species. In both species, a leptobasic type pituitary is found in the ventral zone of the hypothalamus and it is characterized by a neurohypophysis which has a well-developed pituitary stalk and a globular adenohypophysis. The pituitary components, characterized by histochemistry and immunohistochemistry, shows a distribution pattern of cells types similar to other teleost species, with only slight differences in the distribution of βFSH and βLH for P. axelrodi. RESUMEN El cardenal tetra Paracheirodon axelrodi y el tetra Aphyocharax anisitsi son dos especies de carácidos con alto valor comercial como peces ornamentales en América del Sur. Aunque ambas especies habitan en las capas medias de agua, el neón cardenal exhibe una distribución tropical, mientras que el tetra cola roja una distribución subtropical. En general estas especies son difíciles de cultivar, por lo que es esencial conocer algunos componentes clave de los sistemas neuroendocrinos para lograr su reproducción en cautiverio. Considerando la importancia de profundizar en el conocimiento de la fisiología reproductiva a través de la morfología funcional, en este trabajo realizamos, por primera vez, un análisis anatómico, morfológico e inmunohistoquímico de la glándula pituitaria de estas dos especies. En ambas especies, la hipófisis, del tipo leptobásica, se encontró en la zona ventral del hipotálamo y se caracteriza por una neurohipófisis con un tallo hipofisario bien desarrollado y una adenohipófisis globular. Los componentes hipofisarios, caracterizados por la histoquímica y la inmunohistoquímica, mostraron un patrón de distribución de tipos de células similares a otras especies de teleósteos, con solo pequeñas diferencias en la distribución de βFSH y βLH para P. axelrodi.

In Gachantivá, Colombia, Rubus alutaceus has been used for commercial purposes for more than one decade but so far with unknown taxonomic identity. For its identification, the anatomical and morphological characteristics of its flowers, fruits, leaves, stems, and seeds, as well as the physicochemical characteristics of its fruits such as color, equatorial and polar diameter, fresh weight, pH and total soluble solids (TSS), were studied. Qualitative and quantitative morphological parameters were evaluated, and longitudinal and transverse cuts were made for histological studies. The plants were found to have trifoliate and pentafoliate leaves; whitish–yellow subglobose polydrupes, fully mature fruits with 8.8°Brix TSS; elliptical, subglobose seeds with ruminated endocarps; anatomical calcium oxalate crystals in the form of druses and raphides in the leaves and branches; and abundant hairs, simple and glandular (colleters) addition to vascular tissue with a more or less continuous appearance. High value in TSS, positively influences fruit agribusiness and the presence of simple hair and colleters can reduce the attack of pests, these variables identify the new species and give it agronomic potential to strengthen its cultivation and marketing. Resumo No município de Gachantivá, Colômbia, Rubus alutaceus é usada com fins comerciais, ha mais de uma década, sem que até o momento se conheça a sua identidade taxonômica. Com o propósito de identifica-la, estudaram–se as características anatômicas e morfológicas das suas flores, frutos, folhas, caules, sementes; características físico-químicas dos frutos, sua cor, segundo a escala CIELAB, o diâmetro equatorial e polar, peso fresco, pH e sólidos solúveis totais (SST). Avaliaram-se as variáveis morfológicas qualitativas e quantitativas, e realizaram-se cortes longitudinais e transversais com o objetivo de fazer estudos histológicos. Encontraram-se plantas que apresentam folhas trifoliadas e pentafoliadas; polidrupas subglobosas de cor amarela–esbranquiçada, SST de 8,8 °Brix em frutos completamente maduros; sementes elípticas, subglobosas com endocarpo ruminado; anatomicamente apresentam-se cristais de oxalato cálcico em forma de drusas e ráfides nas folhas e ramos e abundantes pelos simples e coléteres, além do tecido vascular de aspecto mais ou menos contínuo. O alto teor em TSS influencia positivamente o agronegócio dos frutos e a presença de estruturas secretoras incluindo tricomas e coléteres simples podem reduzir o ataque de pragas; essas variáveis caracterizam a nova espécie e conferem potencial agronômico para fortalecer seu cultivo e sua comercialização.

Polychaetes are common in coastal and estuarine environments worldwide and constitute one of the most complex groups of marine invertebrates. The morpho-physiology of the female reproductive system (FRS) can be understood by using histological tools to describe reproductive cycle and gametogenesis paths and, among other purposes, aiming to identify and differentiate polychaete species. However, this histology-based approach is rarely combined with molecular tools, which is known to accurately delimitate species. In the same way, the description and understanding of oogenesis and vitellogenesis paths within polychaetes are lacking for most families, narrowing the range of its utility. Therefore, the present study aims to describe the oogenesis in three polychaete species common and abundant on the South American Atlantic coast (Laeonereis culveri, Scolelepis goodbodyi and Capitella biota) and investigate the utility of reproductive features and gametogenesis as a relevant associate knowledge to discriminate species, particularly useful for putative cryptic species, integrated with morphological and molecular data. In a first attempt, the results obtained herein allow the authors to describe two new subtypes of oogenesis, dividing it in extraovarian oogenesis type I and II and intraovarian type I and II. The results also demonstrate that the following histological characters of the FRS can be relevant for the separation of related species: a) oogenesis type, b) occurrence or absence of a true ovary, c) ovary tissue organization, d) type of accessory cells present, and e) oocyte morphology. Additionally, these histological features of FRS, when compared with correlated species studied under this scope, converge with the genetic data. The analysis of cytochrome oxidase I (COI) barcode sequences differentiates between North and South American Atlantic populations of L. culveri (16.78% genetic distance), while in S. goodbodyi and C. biota it discriminates them from their congeneric species. These results highlight the, Os poliquetas são comuns em ambientes costeiros e estuarinos em todo o mundo e constituem um dos grupos mais complexos de invertebrados marinhos. A morfo-fisiologia do sistema reprodutor feminino (FRS) pode ser compreendida por meio de ferramentas histológicas para identificar e diferenciar estes anelídeos. No entanto, essa abordagem histológica raramente é combinada com ferramentas moleculares, amplamente conhecidas por delimitar espécies congenéricas ou crípticas com maior precisão. Do mesmo modo, a descrição e o entendimento da oogênese e vitelogênese dentre os poliquetas, para a maioria das famílias, é ainda limitado. Portanto, o presente estudo tem como objetivo descrever a oogênese em três espécies de poliquetas comuns e abundantes na costa sul-americana (Laeonereis culveri, Scolelepis goodbodyi e Capitella biota) e investigar a utilidade das características reprodutivas e da gametogênese como um conhecimento associado relevante para discriminar espécies, particularmente útil para espécies crípticas putativas, integradas a dados morfológicos e moleculares. Os resultados aqui obtidos permitiram descrever dois novos subtipos de oogênese, dividindo-a em oogênese extra-ovariana dos tipos I e II e intra-ovariana dos tipos I e II. Os resultados também demonstram que os seguintes caracteres histológicos do FRS podem ser relevantes para a separação de espécies relacionadas: a) tipo de oogênese, b) presença ou ausência de um ovário verdadeiro, c) organização tissular ovariana, d) tipo de células acessórias presentes e, e) morfologia do ovócito. Além disso, essas características histológicas do FRS, quando comparadas às espécies correlatas estudadas sob esse escopo, convergem com os dados genéticos separando espécies putativas e congenéricas. As análises com DNA barcode demonstraram que em L. culveri é possível diferenciar as populações atlânticas Norte e Sul-americanas (16,78% de distância genética), enquanto para S. goodbodyi e C. biota fica evidente sua distinção com espécies congenéricas. Esses resultados destacam a importância da abordagem com múltiplas ferramentas e mostram que tanto a histologia quanto a histo-fisiologia do FRS e o DNA barcode podem ser usados para identificar e discriminar espécies crípticas e potencialmente crípticas, o que geralmente não é possível quando se utilizam apenas caracteres morfológicos. Além disso, esses caracteres também podem ser úteis na diferenciação de espécies relacionadas e / ou populações geograficamente distintas desses poliquetas., The authors would like to thank IB/UNICAMP, IO/USP and CEBIMar/USP for providing logistic support. In addition, the authors would like to thank the CBMA and the IB-S for the technical support. This work was supported by the FAPESP (Grants no 2011/50317-5, 2015/25623-6, 2017/06167-5) and CNPq through a productivity grant to A.C.Z.A (306534/2015-0). M.A.L.T was supported by a PhD fellowship (SFRH/BD/131527/2017) from FCT. P.E.V. was supported by a Post-Doctoral Fellowships (BPD1/next-sea/2018, NORTE-01-0145-FEDER-000032). F.O.C. and the University of Minho contribution was supported by the strategic programme UID/BIA/04050/2013 POCI-01-0145-FEDER-007569.

We review recent developments in the understanding of the biomechanics of apicomedial actomyosin and how its contractility can tense and deform tissue. Myosin pulses are driven by a biochemical oscillator but how they are modulated by the mechanical context remains unclear. On the other hand, the emergence of tissue behaviour is highly dependent on the material properties of actin, on how strongly components are connected and on the influence of neighbouring tissues. We further review the use of constitutive equations in exploring the mechanics of epithelial apices dominated by apicomedial Myosin contractility. Sin financiación 5.288 JCR (2018) Q1, 22/174 Genetics & Heredity, 47/193 Cell Biology 3.954 SJR (2018) Q1, 5/87 Developmental Biology, 18/351 Genetics No data IDR 2018 UEM

Objective: To evaluate the neurotrophin mRNA expression and axon count in the median nerve of Wistar rats submitted to neural mobilization (NM) after nerve compression. Methods: Eighteen animals were randomly divided into G1 (nerve compression only), G2 (NM for 1 min), and G3 (NM for 3 min). For NM, the animals were anesthetized and the right scapula received the mobilization, adapted as indicated for humans, on alternate days, from the third to the 13th postoperative (PO) day, totaling six days of therapy. On the 14th PO day, animals were anesthetized and euthanized. Fragments of the median nerve, distal to the compression procedure, were removed for histomorphometric analysis and expression of neurotrophins, nerve growth factor (NGF), and brain-derived neurotrophic factor (BDNF) by RT-PCR. Results: Histomorphometric analysis revealed differences in the number of axons in the injured side, which was significantly lower in the injured limb nerve compared to the control limb, whereas the RT-PCR analysis showed no significant differences in the expression of NGF or BDNF. Conclusion: NM treatment did not affect median nerve regeneration, which maintained normal recovery rates. RESUMO Objetivo: Avaliar a expressão de RNAm de neurotrofinas e a contagem de axônios no nervo mediano de ratos Wistar submetidos à mobilização neural (MN) após compressão nervosa. Métodos: Foram divididos aleatoriamente 18 animais em G1 (apenas compressão nervosa), G2 (MN por 1 minuto) e G3 (MN por 3 minutos). Para a MN, os animais foram anestesiados e o membro escapular direito recebeu a mobilização, adaptada da forma indicada para humanos, em dias alternados, do terceiro ao 13° dia de pós-operatório (PO), em seis dias de terapia. No 14° dia PO, os animais foram anestesiados e eutanasiados. Fragmentos do nervo mediano, distais ao procedimento de compressão, foram retirados para análise histomorfométrica e de expressão das neutrotrofinas, fator de crescimento do nervo (NGF) e fator de crescimento derivado do cérebro (BNDF) por RT-PCR. Resultados: A análise histomorfométrica evidenciou diferenças no número de axônios nos lados lesionados, que foi significativamente menor no nervo do membro lesado comparado com o membro controle; por sua vez, a análise por RT-PCR não apontou diferenças significativas na expressão de NGF e nem de BNDF. Conclusão: O tratamento de MN não afetou a regeneração do nervo mediano, que manteve índices normais de recuperação.

Aedes aegypti (Diptera, Culicidae) is the predominant mosquito vector of several arboviruses such as dengue fever, yellow fever, chikungunya and zika. The larval stage is the major period of feeding and growth of these insects. Many of the products tested for Ae. aegypti control are ingested by the larvae in their natural environment or under laboratory conditions. In some situations, the identification of tissue lesions caused by the ingestion of such products serves as the primary way to understand their mechanism of action and confirm their effectiveness. Therefore, we aimed to conduct studies on the histological and histochemical characteristics of Ae. aegypti larvae in order to contribute to a better understanding of the pathogenicity caused by chemical or biological products for larval control. Therefore, we characterized tissues in the midgut of healthy Ae. aegypti larvae and present high quality images. These images are intended for comparison with results of morphohistochemical studies testing the effect of control substances in mosquito larvae.

Resumen La enfermedad de Hansen (EH) es una infección secundaria a Mycobacterium leprae, de curso crónico y que afecta preferentemente la piel y los nervios periféricos. Es una condición emergente en Chile debido a la migración humana. Presentamos un caso que afectó a un trabajador inmigrante desde Haití, que presentó antes de su arribo múltiples lesiones cutáneas en placas, con hipoestesia y madarosis unilateral. El diagnóstico de la forma multibacilar se estableció por los hallazgos clínicos, la presencia de bacilos ácido-alcohol resistentes en una muestra cutánea y la detección de células inflamatorias alrededor de los terminales nerviosos y granulomas en una biopsia de piel, desde donde se amplificaron segmentos específicos de los genes rpoB y hsp65 de M. leprae. El paciente fue tratado con el esquema combinado estándar de la OMS para la forma multibacilar durante un año, con regresión parcial de las lesiones. Durante el seguimiento se detectó por RPC una disminución del número de copias del gen rpoB de M. leprae en muestras nasofaríngeas hasta el sexto mes. El registro de enfermedades de notificación obligatoria demuestra un incremento reciente de casos de EH en Chile, todos relacionados con población migrante. La enfermedad de Hansen es una nueva condición clínica y los clínicos deben estar atentos a esta posibilidad diagnóstica. Las pruebas moleculares pueden ayudar en el diagnóstico y seguimiento.

Studies on reproduction in sea turtles are important due to its life cycle, migratory patterns, high juvenile mortality and environmental impacts. This study aimed to analyse histomorphometrically gonads of C. mydas from the coastline of the Espírito Santo State, Brazil. Ovaries and testicles were collected between 2014 and 2015 from stranded animals. The material was fixed in formalin 10%, assessed macroscopically and processed for histomorphometrical evaluation. Gonads from 34 individuals were evaluated, twenty-four females and ten males. Macroscopic sexual identification presented 100% accuracy confirmed by histology. Sexual dimorphism was observed in one individual, which was considered as adult (CCL=1.023 m). Microscopy of female gonads revealed predominant previtellogenic follicles; oocyte diameter ranged between 161µm and 750µm and a positive correlation between ovarian length, largest oocyte and CCL was found. In males, autolysis was verified in five individuals. Viable testicles revealed predominant spermatogonia, primary spermatocytes and Sertoli cells in the seminiferous tubules and, Leydig cells and fibroblasts in the stroma. There was a positive correlation between tubular diameter and CCL and testicle length and CCL. Maturation of stromal tissue and a positive correlation between tubular lumen and CCL were also observed. Gonad development is proportional to individual growth. RESUMO Estudos em reprodução de tartarugas marinhas são importantes devido ao ciclo de vida, ao padrão migratório, à alta mortalidade juvenil e aos impactos ambientais. Objetivou-se analisar histomorfometricamente gônadas de C. mydas no litoral do Espírito Santo. Foram coletados ovários e testículos dessa espécie, entre 2014 e 2015. O material foi fixado em formol a 10% e avaliado macroscopicamente. Em seguida, foi processado para avaliação histomorfométrica. Foram avaliadas gônadas de 34 indivíduos, 24 fêmeas e 10 machos. Verificaram-se 100% de acurácia na identificação sexual à macroscopia, confirmada pela histologia. Observou-se dimorfismo sexual em um macho, que foi considerado adulto (CCC=1,023m). A microscopia dos ovários revelou folículos pré-vitelogênicos, cujos ovócitos apresentaram diâmetro médio entre 161µm e 750µm. Houve correlação positiva entre comprimento ovariano e diâmetro do maior ovócito e CCC. Nos machos, verificou-se autólise em cinco indivíduos. Os testículos viáveis revelaram espermatogônias, espermatócitos primários e células de Sertoli nos túbulos seminíferos, além de células de Leydig e fibroblastos no estroma. Houve correlação positiva entre diâmetro tubular e CCC e comprimento testicular e CCC. Verificou-se maturação do tecido estromal e correlação positiva entre o diâmetro do lúmen tubular e o CCC. Verifica-se que o desenvolvimento das gônadas é proporcional ao crescimento dos indivíduos.

We aimed to evaluate the histomorphometry and proteomic profile of the canine uterus during all stages of the reproductive cycle. Eighteen healthy female dogs had their estrous cycle identified by clinical evaluation, vaginal cytology, and serum progesterone levels, which were allocated to the proestrus (n=5), estrus (n=5), diestrus (n=5), and anestrus (n=3) groups. All were submitted to elective ovariosalpingohysterectomy, and the uteri were collected for histomorphometric measurement (Image J software). For proteomic analysis, fragments of the uterine horns were subjected to protein measurement (Bradford method) and extraction by 2D electrophoresis (PDquest software). The results showed that the diestrus promoted greater values of thickness in the uterine structures (μm): uterine wall (2,223.8±229.8), endometrium (819.7±109.1), and myometrium (1,392.6±294.2). Uterus showed a protein profile with good reproducibility per phase (pI: 3.5–9.0; PM: 24–150 KDa), with 11 spots in all phases. Despite the greatest histomorphometric changes in the diestrus, we observed a greater number of spots in the estrus (253±45), followed by the proestrus (185±21), diestrus (113±39), and anestrus (80±21). This finding showed probable participation of these proteins in the uterine preparation for receiving gametes for fertilization. Our results showed greater uterine thickness in the diestrus, and greater protein secretion in the estrus, contributing to the prospection of identification of proteins responsible for the biological reproduction processes. Nuestro objetivo fue evaluar la histomorfometría y el perfil proteómico del útero canino durante todas las etapas del ciclo reproductivo. Se identificó el ciclo estral de dieciocho perras sanas mediante evaluación clínica, citología vaginal y niveles de progesterona sérica, que se asignaron al proestro (n = 5), estro (n = 5), diestro (n = 5) y anestro (n = 3) grupos. Todos se sometieron a una ovariosalpingohisterectomía electiva y se recogieron los úteros para la medición histomorfométrica (software Image J). Para el análisis proteómico, los fragmentos de los cuernos uterinos se sometieron a medición de proteínas (método de Bradford) y extracción por electroforesis 2D (software PDquest). Los resultados mostraron que el diestro promovió mayores valores de espesor en las estructuras uterinas (μm): pared uterina (2.223,8 ± 229,8), endometrio (819,7 ± 109,1) y miometrio (1.392,6 ± 294,2). El útero mostró un perfil proteico con buena reproducibilidad por fase (pI: 3,5-9,0; PM: 24-150 KDa), con 11 manchas en todas las fases. A pesar de los mayores cambios histomorfométricos en el diestro, observamos un mayor número de manchas en el estro (253 ± 45), seguido del proestro (185 ± 21), diestro (113 ± 39) y anestro (80 ± 21). Este hallazgo mostró una probable participación de estas proteínas en la preparación uterina para recibir gametos para la fertilización. Nuestros resultados mostraron mayor grosor uterino en el diestro y mayor secreción de proteínas en el estro, contribuyendo a la prospección de identificación de proteínas responsables de los procesos de reproducción biológica. Nós objetivamos avaliar a histomorfometria e o perfil proteômico do útero canino durante todas as fases do ciclo reprodutivo. Dezoito cadelas saudáveis tiveram seu ciclo estral identificado por avaliação clínica, citologia vaginal e níveis séricos de progesterona, que foram alocados para o proestro (n = 5), estro (n = 5), diestro (n = 5) e anestro (n = 3) grupos. Todas foram submetidas à ovariosalpingohisterectomia eletiva, e os úteros foram coletados para mensuração histomorfométrica (software Image J). Para a análise proteômica, fragmentos das trompas uterinas foram submetidos à dosagem de proteínas (método de Bradford) e extração por eletroforese 2D (software PDquest). Os resultados mostraram que o diestro promoveu maiores valores de espessura nas estruturas uterinas (μm): parede uterina (2.223,8 ± 229,8), endométrio (819,7 ± 109,1) e miométrio (1.392,6 ± 294,2). O útero apresentou um perfil proteico com boa reprodutibilidade por fase (pI: 3,5–9,0; PM: 24–150 KDa), com 11 manchas em todas as fases. Apesar das maiores alterações histomorfométricas no diestro, observamos um maior número de manchas no estro (253 ± 45), seguidas do proestro (185 ± 21), diestro (113 ± 39) e anestro (80 ± 21). Esse achado mostrou provável participação dessas proteínas no preparo uterino para recebimento de gametas para fertilização. Nossos resultados mostraram maior espessura uterina no diestro e maior secreção de proteínas no estro, contribuindo para a prospecção e identificação de proteínas responsáveis ​​pelos processos de reprodução biológica.

Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP) Gillson-Longenbaugh Foundation National Institutes of Health (NIH/NCATS) through the NIH Common Fund, Office of Strategic Coordination (OSC) Extracellular vesicles (EVs) are key mediators of intercellular communication. Part of their biological effects can be attributed to the transfer of cargos of diverse types of RNAs, which are promising diagnostic and prognostic biomarkers. EVs found in human biofluids are a valuable source for the development of minimally invasive assays. However, the total transcriptional landscape of EVs is still largely unknown. Here we develop a new method for total transcriptome profiling of plasma-derived EVs by next generation sequencing (NGS) from limited quantities of patient-derived clinical samples, which enables the unbiased characterization of the complete RNA cargo, including both small- and long-RNAs, in a single library preparation step. This approach was applied to RNA extracted from EVs isolated by ultracentrifugation from the plasma of five healthy volunteers. Among the most abundant RNAs identified we found small RNAs such as tRNAs, miRNAs and miscellaneous RNAs, which have largely unknown functions. We also identified protein-coding and long noncoding transcripts, as well as circular RNA species that were also experimentally validated. This method enables, for the first time, the full spectrum of transcriptome data to be obtained from minute patient-derived samples, and will therefore potentially allow the identification of cell-to-cell communication mechanisms and biomarkers. AC Camargo Canc Ctr, Lab Med Genom, Sao Paulo, SP, Brazil AC Camargo Canc Ctr, Lab Computat Biol, Sao Paulo, SP, Brazil Univ Sao Paulo, Inst Biomed Sci, Dept Cell & Dev Biol, Sao Paulo, SP, Brazil Univ Fed Sao Paulo, Electron Microscopy Ctr, Sao Paulo, SP, Brazil Univ Texas MD Anderson Canc Ctr, Dept Expt Therapeut, Houston, TX 77030 USA Univ Texas MD Anderson Canc Ctr, Ctr RNA Interference & Non Coding RNAs, Houston, TX 77030 USA Univ New Mexico, Comprehens Canc Ctr, Albuquerque, NM 87131 USA Univ New Mexico, Sch Med, Div Hematol Oncol, Dept Internal Med, Albuquerque, NM 87131 USA Univ New Mexico, Sch Med, Div Mol Med, Dept Internal Med, Albuquerque, NM 87131 USA Rockefeller Univ, Lab Mol Immunol, 1230 York Ave, New York, NY 10021 USA FMUSP, Lab Neurociencias Alzira Denise Hertzog Silva LIM, Inst Psiquiatria, Sao Paulo, SP, Brazil Univ Fed Sao Paulo, Electron Microscopy Ctr, Sao Paulo, SP, Brazil FAPESP: 2011/09172-3 FAPESP: 2014/26897-0 Web of Science

Breast cancer is an important public health problem, and its progression may be related to the extracellular matrix (ECM), which acts as a structural scaffold and instruction source for neoplastic cells. Laminins are ECM proteins regulating tumor biology. The laminin-derived peptide C16 regulates different properties of tumor cells. Here we analyzed C16-induced differential gene expression in MDA-MB-231 breast cancer cells. MCF-10A normal-like breast cells served as control. Among different cancer-related genes, C16 induced overexpression of GPNMB. This gene encodes a transmembrane protein GPNMB (glycoprotein non-metastatic B), involved with malignant phenotype of breast cancer cells. Immunoblot validated microarray results. To correlate gene and protein expression with cellular function, we investigated whether C16 would regulate invasion in breast cancer cells. siRNA experiments strongly suggested that C16 and GPNMB cooperate to regulate invasion of highly aggressive MDA-MB-231 cancer cells. We addressed regulatory mechanisms involved in C16-mediated increase of GPNMB protein levels in MDA-MB-231 cells, and observed that C16 stimulates β1 integrin and Src phosphorylation. Furthermore, Src inhibition decreases peptide-induced GPNMB expression levels. To contextualize in vivo our results in vitro, we addressed GPNMB immunostaining in breast cancer human tissue microarrays. Quantitative immunohistochemistry showed that GPNMB is significantly more expressed in breast cancer compared to normal tissue. We concluded that laminin-derived peptide C16 regulates gene and protein expression of GPNMB in breast cancer cells. C16 and GPNMB may cooperate to regulate invasion of highly aggressive MDA-MB-231 cells, probably through Src signaling. GPNMB presented increased expression in breast cancer in vivo compared to normal breast tissue.

Made available in DSpace on 2018-12-11T17:11:37Z (GMT). No. of bitstreams: 0 Previous issue date: 2017-08-01 Astyanax altiparanae is a Brazilian species of substantial commercial, environmental and scientific importance; however, existing studies on its reproduction do not seem to provide enough details. In light of the increasing use of this species in fish farming and the need for basic studies for the development of new production technologies, we describe the structural and ultrastructural characteristics of the ovaries of A. altiparanae, and characterize the species' reproductive cycle. Females were collected monthly from March 2013 to February 2014, and reproductive management began in October 2013. The ovaries were removed, fixed in Karnovsky's fixative, and prepared for light microscopy, transmission electron microscopy and immunohistochemistry anti-PCNA. These techniques enabled us to characterize the ovaries, the germ cells, and the somatic cells in detail, as well as their changes over time. The reproductive cycle was characterized based on the monthly variation in the gonadosomatic ratio, the proportion of germ cells, and the rate of oogonium proliferation. The macroscopic analysis of the ovaries suggests that the vascularization pattern and color of the ovaries vary according to development. There are new types of analyses that can be applied even in the fish farming industry, such as a comparison between ovaries staining and weight or the frequency distribution of these colors throughout the year. This study also provides details on microscopic characteristics that have never before been reported for species of Astyanax, such as the presence of annulate lamellae in oogonia, the development of the zona pellucida from oocytes in the one-nucleolus step, and the development of the micropylar apparatus in oocytes in the cortical alveolar step. When the reproductive cycle was analyzed, this species was found to have a long period of spawning, with a reproductive peak from October to February and multiple spawning events, confirming the period already described in the literature. Variations in reproductive periods and the ability to reproduce in lentic environments suggest that A. altiparanae has the ability to respond quickly to environmental changes and exhibits high reproductive flexibility. All of these characteristics confirm the great potential of this species in the fish farming industry. Fish Endocrinology Laboratory Department of Cell and Developmental Biology Institute of Biomedical Sciences University of Sao Paulo, Av. Prof Lineu Prestes n. 1524, lab 426 Hydrobiology and Aquaculture Station Sao Paulo Energy Company (CESP), Rod. Tamoios n. 21500 Fishery Institute APTA SAA, Av. Francisco Matarazzo n.455 Department of Morphology Biosciences Institute UNESP – Botucatu District of Rubião Junior s/n CEP Department of Morphology Biosciences Institute UNESP – Botucatu District of Rubião Junior s/n CEP

Visualization and clear understanding of the ovarian structures are important in determining the stage of oestrus, helping to diagnose several pathologies and supporting advances in reproductive technologies. In this research, computerized microtomography (microCT) was used to explore and characterize the ovarian structure of seven mammalian species. Ovaries of rats, female dog, queens, cows, mares, sows and a female donkey were used. After microCT scanning, the same samples were prepared for histologic evaluation, used here as a validation criterion. It was possible to distinguish regions of the cortex and medulla, visualize the morphology and distribution of blood vessels, clearly observe corpus luteum and antral follicles, and visualize oocytes inside some antral follicles. This is the first report using microCT to explore and compare ovarian structures in several domestic mammals. MicroCT revealed great potential for the evaluation of ovarian structures. This research open prospects for the use of computerized tomography (CT) as a non-invasive approach to studying ovarian structures in live animals, which may be especially attractive for scientific study of development of ovarian structures and/or ovarian pathologies in small animals’ models.

Federal University of Para. Institute of Biological Sciences. Laboratory of Parasitology and Laboratory of Structural Biology. Bel?m, PA, Brazil / National Institute of Science and Technology in Structural Biology and Bioimaging. Rio de Janeiro, RJ, Brazil. Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas. Ananindeua, PA, Brasil / National Institute of Science and Technology in Structural Biology and Bioimaging. Rio de Janeiro, RJ, Brazil. Federal University of Para. Institute of Biological Sciences. Laboratory of Parasitology and Laboratory of Structural Biology. Bel?m, PA, Brazil. University of S?o Paulo. Department,Institute of Biomedical Sciences. Cell and Developmental Biology. S?o Paulo, SP, Brazil. Butantan Institute. Laboratory of Pathophysiology. S?o Paulo, SP, Brazil / University of S?o Paulo. Institute of Biomedical Sciences. Department of Pharmacology. S?o Paulo, SP, Brazil. Federal University of Para. Institute of Biological Sciences. Laboratory of Parasitology and Laboratory of Structural Biology. Bel?m, PA, Brazil / National Institute of Science and Technology in Structural Biology and Bioimaging. Rio de Janeiro, RJ, Brazil. American tegumentary leishmaniasis is caused by different species of Leishmania. This protozoan employs several mechanisms to subvert the microbicidal activity of macrophages and, given the limited efficacy of current therapies, the development of alternative treatments is essential. Animal venoms are known to exhibit a variety of pharmacological activities, including antiparasitic effects. Crotoxin (CTX) is the main component of Crotalus durissus terrificus venom, and it has several biological effects. Nevertheless, there is no report of CTX activity during macrophage - Leishmania interactions. Thus, the main objective of this study was to evaluate whether CTX has a role in macrophage M1 polarization during Leishmania infection murine macrophages, Leishmania amazonensis promastigotes and L. amazonensis-infected macrophages were challenged with CTX. MTT [3-(4,5dimethylthiazol-2-yl)-2,5-diphenyl tetrasodium bromide] toxicity assays were performed on murine macrophages, and no damage was observed in these cells. Promastigotes, however, were affected by treatment with CTX (IC50 = 22?86 ?g mL-1) as were intracellular amastigotes. Macrophages treated with CTX also demonstrated increased reactive oxygen species production. After they were infected with Leishmania, macrophages exhibited an increase in nitric oxide production that converged into an M1 activation profile, as suggested by their elevated production of the cytokines interleukin-6 and tumour necrosis factor-? and changes in their morphology. CTX was able to reverse the L. amazonensis-mediated inhibition of macrophage immune responses and is capable of polarizing macrophages to the M1 profile, which is associated with a better prognosis for cutaneous leishmaniasis treatment.

Purpose We have previously shown that the development of complications in the early pregnant decidua and myometrium in mice correlates with diabetes progression. In the current study, we investigated the influence of diabetes progression on the placental extracellular matrix (ECM) and on fetal development at the end of pregnancy. Methods Alloxan-induced type 1 diabetic female mice were bred either 30–50 days after diabetes induction (D) or 90-110D. Fetal and placental weights were registered at the 19th day of pregnancy together with analysis of gene expression, deposition and turnover of the placental ECM. Results The short-term diabetic group (30-50D) showed elevated embryonic losses and underweight fetuses (89%) with normal weight placentas. In contrast, the long-term group (90-110D) had increased malformations/fetal deaths and underweight fetuses (42%) and heavy placentas (50%). Normal-weight fetuses from the long-term group had placentas with either regular weight and fetal/placental weight ratio or increased weight and low fetal/placental weight ratio. Furthermore, the placentas of the short-term group showed alterations in the synthesis and deposition of collagen types I and V and in the activity of MMP2 whereas placentas of the 90-110D group presented alterations in collagen type III and V and MMP9. Conclusions Diabetes progression promoted distinct outcomes in pregnancy. Modifications of both synthesis and turnover of ECM occurred even before changes of placental weight were detected. Adjustment of fetal/placental weight ratio or placental enlargement restored normal growth in part of the fetuses from the long-term group.

Recent studies seem to indicate that apoptosis and autophagy can act cooperatively in fish ovaries in order to achieve more effective ovarian regression after spawning. Considering the importance of tissue remodeling in ovarian functioning, we sought to morphologically characterize the involution processes of follicular atresia and post-ovulatory complexes using Astyanax altiparanae as an experimental model, and to determine the location of proteins involved in apoptosis and autophagy throughout this process. Fifteen females were collected after reproductive management. Fragments of the left ovaries were removed, fixed, and prepared for light microscopy and immunofluorescence analyses. The main characteristics of the involution processes were found to be consistent with previous descriptions. However, there were certain morphological peculiarities that do not appear to have been described for any other species thus far. These peculiarities may be related to the focus of this study on a single species, which allows for a more detailed investigation into morphological changes than studies on multiple species. Autophagy was also found to precede apoptosis in both involution processes in A. altiparanae. This may be related to the energy recycling process required before the removal of degenerated follicular cells by apoptosis. Thus, these results support the idea that there is crosstalk between autophagy and apoptosis pathways in ovarian involution processes, as well as the idea that the cell death pathways of these processes are conserved between teleost species with external fertilization.

Background Glioblastoma (GBM), a highly aggressive brain tumor, contains a subpopulation of glioblastoma stem-like cells (GSCs) that play roles in tumor maintenance, invasion, and therapeutic resistance. GSCs are therefore a promising target for GBM treatment. Our group identified the cellular prion protein (PrPC) and its partner, the co-chaperone Hsp70/90 organizing protein (HOP), as potential target candidates due to their role in GBM tumorigenesis and in neural stem cell maintenance. Methods GSCs expressing different levels of PrPC were cultured as neurospheres with growth factors, and characterized with stem cells markers and adhesion molecules markers through immunofluorescence and flow cytometry. We than evaluated GSC self-renewal and proliferation by clonal density assays and BrdU incorporation, respectively, in front of recombinant HOP treatment, combined or not with a HOP peptide which mimics the PrPC binding site. Stable silencing of HOP was also performed in parental and/or PrPC-depleted cell populations, and proliferation in vitro and tumor growth in vivo were evaluated. Migration assays were performed on laminin-1 pre-coated glass. Results We observed that, when GBM cells are cultured as neurospheres, they express specific stemness markers such as CD133, CD15, Oct4, and SOX2; PrPC is upregulated compared to monolayer culture and co-localizes with CD133. PrPC silencing downregulates the expression of molecules associated with cancer stem cells, upregulates markers of cell differentiation and affects GSC self-renewal, pointing to a pivotal role for PrPC in the maintenance of GSCs. Exogenous HOP treatment increases proliferation and self-renewal of GSCs in a PrPC-dependent manner while HOP knockdown disturbs the proliferation process. In vivo, PrPC and/or HOP knockdown potently inhibits the growth of subcutaneously implanted glioblastoma cells. In addition, disruption of the PrPC-HOP complex by a HOP peptide, which mimics the PrPC binding site, affects GSC self-renewal and proliferation indicating that the HOP-PrPC complex is required for GSC stemness. Furthermore, PrPC-depleted GSCs downregulate cell adhesion-related proteins and impair cell migration indicating a putative role for PrPC in the cell surface stability of cell adhesion molecules and GBM cell invasiveness, respectively. Conclusions In conclusion, our results show that the modulation of HOP-PrPC engagement or the decrease of PrPC and HOP expression may represent a potential therapeutic intervention in GBM, regulating glioblastoma stem-like cell self-renewal, proliferation, and migration. Electronic supplementary material The online version of this article (doi:10.1186/s13287-017-0518-1) contains supplementary material, which is available to authorized users.

O objetivo deste trabalho foi analisar a morfologia dos testículos de tuviras machos (Gymnotus spp.) submetidas à reprodução por diferentes indutores hormonais. Foram utilizadas quarenta tuviras macho de aproximadamente 40 g em um delineamento inteiramente casualizado, com quatro tratamentos e 10 repetições, sendo que cada animal foi considerado uma repetição. Para estabelecimento do protocolo hormonal foram testados três indutores: extrato bruto de hipófise de carpa (EBHC), gonadotrofina coriônica equina (ECG), análogo sintético do hormônio liberador de gonadotrofina (GnRH-a) e no grupo controle foi administrada solução salina a 2%. Os três hormônios e o controle foram aplicados em duas dosagens (preparatória e indução final). Após 12 horas da segunda aplicação, cinco machos de cada tratamento foram eutanasiados para avaliação macroscópica e microscópica. Foi determinada a proporção volumétrica ocupada pelos componentes do parênquima testicular (túbulos seminíferos e tecido intertubular) e quantificadas as populações de espermatogônias do tipo A, do tipo B, espermatócitos primários, espermatócitos secundários, espermátides, células de Sertoli (S) e proporção de Lúmen. Os resultados indicaram que o hormônio GnRH-a apresentou uma maior capacidade para promover a maturação final nos testículos, seguido pelo hormônio eCG. O hormônio EBHC não conseguiu promover a maturação final nas tuviras induzidas.

Objective In this study we performed a temporal analysis of the effects of Diabetes Mellitus on morphology and laminin deposition in salivary glands of young (2 months-old) and aging (12 months-old) male Wistar rats, using immunohistochemistry. Materials and methods The animals were divided in control and diabetic (Streptozotocin induced) groups and euthanized after short and long-term diabetes induction. Results Short-term induction led to vacuolization of parotid acinar cells and increased laminin deposition in both animal ages. In young rats, no difference was observed between short or long-term diabetes regarding laminin deposition, but parotid acinar cells vacuolization was more discrete after long-term diabetes. A slight decrease of submandibular gland convoluted granular ducts was observed in young and elder diabetic animal ages. In diabetic aging rats was observed an increase of laminin content only in the parotid gland. Conclusions These results suggest that some Diabetes Mellitus effects on salivary glands are not progressive over time, possibly due to the existence of adaptive mechanisms in response to chronic hyperglycemia. They also show that the duration of the disease was more relevant to the morphological effects than the age, although it is known that aging per se affects salivary gland morphology and function.

The nurse shark, Ginglymostoma cirratum (Bonnaterre, 1778) is one of the most studied species of elasmobranchs. However, the knowledge of their reproductive biology is still relatively rare, particularly in the western South Atlantic. This study aimed to describe the morphology of the uterus and the ovary of G. cirratum, based on specimens caught off at the Fortaleza/CE coast, northeast Brazil. Samples were collected from September 2012 to June 2013, from regular landings of artisanal fishing, which commercialize this species freely. A total of ten females were collected. The methodologies followed for analyzing the ovaries and uterus of those females included both macroscopic and histological analysis. G. cirratum has internal type ovary morphology, with invaginations of connective tissue, which defines compartments and separate oocyte groups in ovigerous lots. The epithelium lining the ovary changes from simple columnar ciliated in the area without ovigerous lots, which turns into a simple cubic epithelium in the coating portion of the epigonal organ where ovarian tissue is absent. The uterine mucosa has secretory cells denoted by Alcian Blue staining, indicating the production of mucopolysaccharides, even in immature individuals. This lecithotrophic shark has a uterine vascularized mucosa that is one characteristic of viviparous elasmobranch species. RESUMO: O tubarão-lixa, Ginglymostoma cirratum (Bonnaterre, 1778) é uma das espécies mais estudadas de elasmobrânquios. O conhecimento de sua biologia reprodutiva, no entanto, ainda é relativamente raro, particularmente no Oeste do Atlântico Sul. Este estudo objetivou a descrição morfológica do útero e ovário de G. cirratum, baseado em espécimes capturados na costa de Fortaleza/CE, no Nordeste do Brasil. Um total de dez fêmeas foi coletado de setembro de 2012 a junho de 2013, nos desembarques regulares da pesca artesanal, que comercializam essa espécie livremente. A metodologia foi de acordo com as analises macroscópica e histológica dos ovários e útero. G. cirratum tem o tipo de morfologia de ovário interno, com invaginações de tecido conjuntivo, que define compartimentos e separa grupos de oócitos em lotes ovígeros. O epitélio que reveste o ovário varia de epitélio simples colunar ciliado na área que não apresenta lotes ovígeros para epitélio simples cúbico na porção de revestimento do órgão epigonal, onde o tecido ovariano é ausente. A mucosa uterina possui células secretoras, evidenciadas pela coloração em Alcian Blue, indicando a produção de mucopolissacarídeos, mesmo em indivíduos imaturos. Este tubarão lecitotrófico apresenta a mucosa uterina vascularizada, sendo esta uma característica de espécies de elasmobrânquios vivíparos.

Objective To evaluate the β-hydroxy-βmethylbutyrate supplementation influence on the morphological and morphometric characteristics of the diaphragm muscle of rats and verify whether there are sex differences. Methods Experimental study with 48 Wistar rats (24 of each sex) divided into 3 groups: Control Group: in which a daily diet with saline solution was offered; Experimental Group: the same amount of food from the Control group consumed in the previous day and 0.3g/kg of β-hydroxy-βmethylbutyrate; Ad libitum Experimental Group: ad libitum feeding with the same dose of the supplement. The analysis consisted of histomorphometry and classification in diaphragm muscle fiber area bands. The procedures occurred 30 days after the start of the experiment. Data were analyzed using the one-way ANOVA and Tukey tests (p

Stable isotope analysis of baleen plates is a widespread technique for studying baleen whales. Typically, subsamples along the growth axis of the baleen plate are extracted and analysed to examine time-related variation in their stable isotope signals. However, baleen plate tissue is composed of two different tissues: a pair of cortex layers flanking an internal medulla. These two histological components exhibit differential development, and their consolidation as a tissue is therefore likely non-synchronic. This could influence stable isotope results because the stable isotope signal may differ in each subsample according to the proportion of the two histological components extracted from the tissue. In this study, stable isotope analysis was combined with optical microscopy examination of fin whale (Balaenoptera physalus) baleen plates to understand the ontogeny of the two histological components. In both of them, the δ15N values followed a sinewave pattern along the growth axis of the baleen plate. However, the δ15N values of the cortex appeared to be advanced compared to those of the medulla. Additionally, the amplitude of the δ15N values in the oscillations was higher in the cortex than in the medulla. The histological examination revealed that these differences are caused by earlier and faster synthesis of the cortex layer compared to that of the medulla. Because the stable isotope ratios of the two layers differ, we propose that in this type of studies only the outer-most part (closest to the surface) of the cortex should be subsampled and analysed. Additionally, to include the most recently formed tissue, this subsampling should start well below the zwischensubstanz, or baleen “gum”.

Macrobrachium amazonicum (Heller, 1862) is the most important species for aquaculture native to South America. This study evaluates the phenotypic plasticity of females of Macrobrachium amazonicum with respect to the gonadal development, and determines the spawning type. Our study consisted of macro and microscopic analysis of the reproductive tract. Ovaries exhibited the following reproductive cells in developmental sequence: oogonia (OO) - mean diameter and standard deviation of 25.4 ± 6.5 μm; previtellogenic oocyte (PVO) - 61.7 ± 10.7 μm, vitellogenic oocyte (VO) - 113.9 ± 24.5 μm; and mature oocyte (MO) - 308.7 ± 56.3 μm. Ovaries increased in volume due to vitellogenesis and changing of basophilic to acidophilic composition. Follicular cells, atresic oocytes and postovulatory follicles were also analyzed. By combining macro and microscopic analysis, the ovaries of M. amazonicum were classified into six stages: Immature, In maturation, Mature, Spawned in maturation, Spawned and Resting. The ovarian development of M. amazonicum followed a standard pattern even among different populations. Considering our results and the evidence in literature, we conclude that spawning for this species is parceled or multiple, that is, synchronous in more than two groups (clutches of oocytes). Such observations provide basis for further studies addressing sustainable management strategies for species conservation and contribute to elucidate the biology of their specimens. RESUMO Das espécies nativas de camarões da América do Sul, Macrobrachium amazonicum (Heller, 1862) é a mais importante para fins de aquicultura, possuindo elevado valor comercial. Devido à sua importância, este estudo objetivou avaliar a plasticidade fenotípica de fêmeas da espécie em relação ao desenvolvimento gonadal, e determinar o tipo de desova. Para tal, foi realizada análise macro e microscópica do aparelho reprodutor. Observaram-se ovários apresentando as seguintes células reprodutivas em sequência de desenvolvimento: oogônias (OO) com diâmetro médio e desvio padrão de 25,4 ± 6,5 µm, oócito pré-vitelogênico (OPV) com 61,7 ± 10,7 µm, oócito vitelogênico (OV) com 113,9 ± 24,5 µm, e oócito maturo (OM) com 308,7 ± 56,3 µm, havendo um aumento do volume devido ao processo de vitelogênese e uma alteração de composição basófila para acidófila. Além destas células, observaram-se células foliculares, oócitos atrésicos e folículos pós-ovulatórios. Unindo análise macro e microscópica, os ovários de M. amazonicum puderam ser classificados em seis estágios: Imaturo, Em maturação, Maturo, Desovado em maturação, Desovado e Repouso. Verificou-se que o desenvolvimento ovariano de M. amazonicum segue um padrão para as diferentes populações. Diante dos resultados encontrados e evidências na literatura, constata-se que a desova para esta espécie é do tipo parcelada ou múltipla, ou seja, sincrônica em mais de dois grupos (lotes de oócitos). Estas observações oferecem uma base para estudos que contribuirão para a criação de medidas de manejo sustentáveis para a manutenção da espécie na natureza, bem como contribuem para elucidar a biologia de seus indivíduos.

Chromoblastomycosis (CBM) is a cutaneous or subcutaneous mycoses. The trauma occurs when the fungus is installed and is more prevalent in individuals living in tropical and subtropical regions, with earliest descriptions dating back to 1920. The diagnosis of CBM is based on the incidence of cases in the endemic areas and is commonly reached through microbiological analyses to identify the etiologic agent in clinical samples. The process for the analysis of the collected samples allows one to visualise the muriform cells, which are brown, rounded structures having crossed chambers and that can be commonly called sclerotic bodies, characterising the positive diagnosis. The objective of this review was to verify the connection of the histopathological techniques to the diagnosis of CBM. La cromoblastomicosis (CBM) es una micosis cutánea o subcutánea. El trauma ocurre cuando el hongo se establece y es más prevalente en individuos que viven en regiones tropicales y subtropicales; las primeras descripciones datan de 1920. El diagnóstico de CBM se basa en la incidencia de casos en áreas endémicas y comúnmente se logra a través de medios microbiológicos. análisis para identificar el agente etiológico en muestras clínicas. El proceso de análisis de las muestras recolectadas permite visualizar las células muriformes, que son estructuras marrones, redondeadas, con cámaras cruzadas y que comúnmente pueden denominarse cuerpos escleróticos, caracterizando el diagnóstico positivo. El propósito de esta revisión fue verificar la conexión entre las técnicas histopatológicas y el diagnóstico de CBM. A cromoblastomicose (CBM) é uma micose cutânea ou subcutânea. O trauma ocorre quando o fungo se instala e é mais prevalente em indivíduos que vivem em regiões tropicais e subtropicais, com as primeiras descrições datando de 1920. O diagnóstico de CBM é baseado na incidência de casos em áreas endêmicas e é comumente alcançado por meio microbiológico análises para identificar o agente etiológico em amostras clínicas. O processo de análise das amostras coletadas permite visualizar as células muriformes, que são estruturas castanhas, arredondadas, com câmaras cruzadas e que podem ser comumente chamadas de corpos escleróticos, caracterizando o diagnóstico positivo. O objetivo desta revisão foi verificar a conexão das técnicas histopatológicas com o diagnóstico de CBM.

Honeybees are the insect most used for pollination purposes due to its efficient characteristics for this function, which reflects in positive aspects for both nature and man. The expansion of agriculture and the development of agrochemicals to combat pests has had negative impacts on honeybee health, causing its disappearance around the world. This research aimed to evaluate the effects of honeybee exposure to the neonicotinoid insecticide thiamethoxam on its survival rate, as well as on morphological and histological changes in the midgut of adult workers from the F4 generation of Apis mellifera queens tolerant to thiamethoxam and non-tolerant worker honeybees. After the bioassays, the midgut was removed for morphological evaluation. The results showed that F4 bees were more tolerance to thiamethoxam and exhibited less significant morphological changes when compared to the non-tolerant control group. Las abejas son el insecto más utilizado con fines de polinización por sus características eficientes para esta función, lo que se refleja en aspectos positivos tanto para la naturaleza como para el hombre. La expansión de la agricultura y el desarrollo de agroquímicos para combatir plagas ha tenido impactos negativos en la salud de las abejas, provocando su desaparición en todo el mundo. Esta investigación tuvo como objetivo evaluar los efectos de la exposición de las abejas melíferas al insecticida neonicotinoide tiametoxam sobre su tasa de supervivencia, así como sobre los cambios morfológicos e histológicos en el intestino medio de trabajadores adultos de la generación F4 de reinas Apis mellifera tolerantes al tiametoxam y trabajadoras no tolerantes abejas. Después de los bioensayos, se extrajo el intestino medio para evaluación morfológica. Los resultados mostraron que las abejas F4 eran más tolerantes al tiametoxam y exhibían cambios morfológicos menos significativos en comparación con el grupo de control no tolerante. A abelha melífera é o inseto mais utilizado para fins de polinização devido às suas características eficientes para esta função, o que se reflete em aspectos positivos tanto para a natureza quanto para o homem. A expansão da agricultura e o desenvolvimento de agroquímicos para combater as pragas tiveram impactos negativos na saúde das abelhas, causando seu desaparecimento em todo o mundo. Esta pesquisa teve como objetivo avaliar os efeitos da exposição das abelhas ao inseticida neonicotinoide tiametoxam sobre sua taxa de sobrevivência, bem como sobre as alterações morfológicas e histológicas do intestino médio de operárias adultas da geração F4 de Apis mellifera rainhas tolerantes ao tiametoxam e trabalhadoras não tolerantes abelhas. Após os bioensaios, o intestino médio foi removido para avaliação morfológica. Os resultados mostraram que as abelhas F4 foram mais tolerantes ao tiametoxam e exibiram alterações morfológicas menos significativas quando comparadas ao grupo controle não tolerante.

The pampas cat Leopardus colocolo (Molina, 1782) is a species of the Felidae family, widely distributed in South America, included on CITES Appendix II and classified as Near Threatened on the IUCN Red List, with population trend decreasing. Based on this information, the objective of this study is to describe morphologically the testes and epididymal ducts of pampas cat. The animal, coming from the Federal University of Mato Grosso Zoo, Brazil, had died after anesthesia procedure and the male reproductive system was dissected to collect the testicles. The samples taken were fragmented and histologically examined. From the microscopic analysis of the testes were identified: vaginal and tunica albuginea, formed by dense connective tissue modeled with large amount of collagen fibers. The tunica albuginea fibrous septa emits into the body. The seminiferous tubules are coiled and coated internally by spermatogenic epithelium consisting of Sertoli cells, surrounded by a basement membrane in the presence of myoid cells. The interstitial tissue between the seminiferous tubules, is composed of loose connective tissue, blood and lymph vessels, and Leydig cells in polyhedral shape. The epididymal ducts showed pseudostratified columnar epithelium with secretory cells of which stereocilia design, situated on a basement membrane filled by myoid cells. This epithelium has principal and basal cells, the main cell design stereocilia toward the lumen of the epididymal duct.