Opportunistic meat-eating by urban folivorous-frugivorous monkeys.

The consumption of vertebrate tissues and eggs (hereinafter "meat") is relatively common among some primates that are highly frugivorous or eclectic omnivores, but rare or absent in those that are highly folivorous. The Neotropical howler monkeys (Alouatta spp.) belong in the latter group. Here we report the consumption of meat by free-ranging urban black and gold howler monkeys (Alouatta caraya) and discuss the potential role of the consumed meat as a source of energy, protein, or micronutrients. We studied three groups of howler monkeys (comprising four to seven individuals), living in city squares (0.6, 1.5, and 1.9 ha) in south Brazil, from July 2022 to May 2023 (65 days; 797 h of observations). All of the study groups were spontaneously supplemented daily by people with variable amounts and types of food provided. Meat was only offered in the two larger squares. The groups' diets included leaves (42-49% scan sampling feeding records), fruit (3-20%), and flowers (2-5%) from 13 to 20 plant species, and considerable amounts of supplemented food (27-50%). We recorded 33 individual events of ingestion of supplemented cooked meat, three individual events of dove egg predation, and three bird nest inspections without egg consumption. All members of the two groups in the larger squares, except an infant male, ingested meat at least once. Meat accounted for 1% of total scan feeding records of both groups with access to this supplement. We conclude that whereas the opportunistic consumption of meat probably contributed only minor amounts of energy and protein to the study subjects, it may have benefitted them with micronutrients that are scarce in plant foods.

Application of artificial intelligence in predicting malignancy risk in breast masses on ultrasound / Aplicação de inteligência artificial na predição de risco de malignidade em nódulos mamários na ultrassonografia

Abstract Objective: To evaluate the results obtained with an artificial intelligence-based software for predicting the risk of malignancy in breast masses from ultrasound images. Materials and Methods: This was a retrospective, single-center study evaluating 555 breast masses submitted to percutaneous biopsy at a cancer referral center. Ultrasonographic findings were classified in accordance with the BI-RADS lexicon. The images were analyzed by using Koios DS Breast software and classified as benign, probably benign, low to intermediate suspicion, high suspicion, or probably malignant. The histological classification was considered the reference standard. Results: The mean age of the patients was 51 years, and the mean mass size was 16 mm. The radiologist evaluation had a sensitivity and specificity of 99.1% and 34.0%, respectively, compared with 98.2% and 39.0%, respectively, for the software evaluation. The positive predictive value for malignancy for the BI-RADS categories was similar between the radiologist and software evaluations. Two false-negative results were identified in the radiologist evaluation, the masses in question being classified as suspicious by the software, whereas four false-negative results were identified in the software evaluation, the masses in question being classified as suspicious by the radiologist. Conclusion: In our sample, the performance of artificial intelligence-based software was comparable to that of a radiologist.

Resumo Objetivo: O objetivo deste trabalho foi avaliar os resultados de um software baseado em algoritmo de inteligência artificial para predição do risco de malignidade em nódulos mamários. Materiais e Métodos: Estudo retrospectivo e unicêntrico que avaliou 555 nódulos mamários submetidos a biópsia percutânea em um centro de referência oncológico. Os achados ultrassonográficos foram classificados de acordo com o léxico do BI-RADS. As imagens foram analisadas pelo software Koios DS Breast e divididas em benigna ou provavelmente benigna, suspeita baixa ou intermediária, suspeita alta ou provavelmente maligna. O resultado histopatológico foi considerado como padrão ouro. Resultados: A média de idade das pacientes foi de 51 anos e o tamanho médio dos nódulos foi de 16 mm. A sensibilidade e a especificidade foram de 99,1% e 34,0% para o radiologista e 98,2% e 39,0% para o software, respectivamente. O valor preditivo positivo para malignidade para as categorias BIRADS foi semelhante para o radiologista e para o software. Foram identificados dois resultados falso-negativos na avaliação pelo radiologista que foram classificados como suspeitos pelo software, e quatro resultados falso-negativos na avaliação pelo software que foram classificados como suspeitos pelo radiologista. Conclusão: Na nossa amostra, o software de inteligência artificial demonstrou resultados comparáveis à avaliação pelo radiologista.

Application of artificial intelligence in predicting malignancy risk in breast masses on ultrasound.

Objective: To evaluate the results obtained with an artificial intelligence-based software for predicting the risk of malignancy in breast masses from ultrasound images. Materials and Methods: This was a retrospective, single-center study evaluating 555 breast masses submitted to percutaneous biopsy at a cancer referral center. Ultrasonographic findings were classified in accordance with the BI-RADS lexicon. The images were analyzed by using Koios DS Breast software and classified as benign, probably benign, low to intermediate suspicion, high suspicion, or probably malignant. The histological classification was considered the reference standard. Results: The mean age of the patients was 51 years, and the mean mass size was 16 mm. The radiologist evaluation had a sensitivity and specificity of 99.1% and 34.0%, respectively, compared with 98.2% and 39.0%, respectively, for the software evaluation. The positive predictive value for malignancy for the BI-RADS categories was similar between the radiologist and software evaluations. Two false-negative results were identified in the radiologist evaluation, the masses in question being classified as suspicious by the software, whereas four false-negative results were identified in the software evaluation, the masses in question being classified as suspicious by the radiologist. Conclusion: In our sample, the performance of artificial intelligence-based software was comparable to that of a radiologist.

Objetivo: O objetivo deste trabalho foi avaliar os resultados de um software baseado em algoritmo de inteligência artificial para predição do risco de malignidade em nódulos mamários. Materiais e Métodos: Estudo retrospectivo e unicêntrico que avaliou 555 nódulos mamários submetidos a biópsia percutânea em um centro de referência oncológico. Os achados ultrassonográficos foram classificados de acordo com o léxico do BI-RADS. As imagens foram analisadas pelo software Koios DS Breast e divididas em benigna ou provavelmente benigna, suspeita baixa ou intermediária, suspeita alta ou provavelmente maligna. O resultado histopatológico foi considerado como padrão ouro. Resultados: A média de idade das pacientes foi de 51 anos e o tamanho médio dos nódulos foi de 16 mm. A sensibilidade e a especificidade foram de 99,1% e 34,0% para o radiologista e 98,2% e 39,0% para o software, respectivamente. O valor preditivo positivo para malignidade para as categorias BIRADS foi semelhante para o radiologista e para o software. Foram identificados dois resultados falso-negativos na avaliação pelo radiologista que foram classificados como suspeitos pelo software, e quatro resultados falso-negativos na avaliação pelo software que foram classificados como suspeitos pelo radiologista. Conclusão: Na nossa amostra, o software de inteligência artificial demonstrou resultados comparáveis à avaliação pelo radiologista.

Hematological Changes in Dogs with Visceral Leishmaniasis Are Associated with Increased IFN-γ and TNF Gene Expression Levels in the Bone Marrow.

Visceral leishmaniasis is associated with a variety of hematological abnormalities. In this study, we correlated the hematological changes in the peripheral blood of dogs naturally infected with Leishmania infantum (L. infantum) with the distribution of cell lineages and cytokine gene expression patterns in the bone marrow. Samples from 63 naturally semidomiciled dogs living in an endemic area of visceral leishmaniasis were analyzed. L. infantum infection was detected in 50 dogs (79.3%). Among those, 18 (32%) had positive splenic cultures and showed more clinical signs. They also had lower red blood cell counts and leukocytosis with an increased number of neutrophils and monocytes in peripheral blood compared to dogs negative to this test. L. infantum DNA was detected in the bone marrow of 8/14 dogs with positive splenic culture. Dogs with L. infantum infection in the bone marrow presented with histiocytosis (p = 0.0046), fewer erythroid cell clusters (p = 0.0127) and increased gene expression levels of IFN-γ (p = 0.0015) and TNF (p = 0.0091). The data shown herein suggest that inflammatory and cytokine gene expression changes in bone marrow may contribute to the peripheral blood hematological changes observed in visceral leishmaniasis.

Placenta percreta com invasão de bexiga: um relato de caso / Percretal placenta with bladder invasion: a case report

O acretismo placentário consiste na aderência anormal da placenta na parede uterina. Ao aderir-se diretamente ao miométrio, denomina-se placenta acreta; ao estender-se mais profundamente, placenta increta, e ao invadir a serosa uterina ou órgãos adjacentes, percreta. O fator de risco mais frequente constitui cesarianas anteriores. Paciente 27 anos, G3P1CA1 (cesariana há 8 anos/ parto prematuro 25 sem há 4 anos), IG: 25sem3d; com alteração da vitalidade fetal e placenta prévia com sinais de acretismo (sugerindo placenta percreta). Foi indicada a interrupção da gestação com 27 semanas e 1 dia. No período intraoperatório foi evidenciada, por meio de ultrassom, presença de acretismo placentário com invasão miometrial e invasão de serosa vesical sendo posteriormente realizado a histerectomia subtotal e rafia das lacerações da mucosa vesical. A placenta percreta é mais frequente em grávidas com placenta prévia no local da cicatriz de cesariana e o órgão mais frequentemente acometido é a bexiga; estando associada a maior morbimortalidade materna. O diagnóstico definitivo é anatomopatológico, porém é presumível durante a cirurgia abdominal com a visualização da invasão placentária, devendo ser confirmado por Histopatologia.

Placental accretism consists of abnormal placental adherence to the uterine wall. When adhering directly to the myometrium it is called placenta accreta; when extending more deeply, placenta increta; and when invading the uterine serosa or adjacent organs, percrete. The most frequent risk factor is previous cesarean sections. The patient is 27 years old with altered fetal vitality and placenta previa with signs of accreation (suggesting percretal placenta). Pregnancy termination at 27 weeks and one day was indicated. In the intraoperative period, the presence of placental accretion with myometrial invasion and bladder serous invasion was evidenced by ultrasound, with subtotal hysterectomy and raffia of lacerations of the bladder afterwards. The percretal placenta is more frequent in pregnant women with placenta previa at the site of the scar of a cesarean section and the organ most frequently affected is the bladder; being associated with higher maternal morbidity and mortality. The definitive diagnosis is anatomopathological, but it is presumed during abdominal surgery with the visualization of the placental invasion and must be confirmed by Histopathology

Sexualidade na terceira idade e educação em saúde: um relato de experiência / Sexuality in old age and health education: an experience report

A pirâmide etária mostra um envelhecimento progressivo no mundo. Esse processo envolve aspectos relacionados à saúde, entre eles a sexualidade como variável interferente na qualidade de vida do ser humano. Este relato buscou discorrer sobre uma ação de educação em saúde voltada para os idosos, realizada no âmbito da Atenção Primária em Saúde, desenvolvida por estudantes de medicina. A experiência ocorreu por meio de rodas de conversa e aconselhamento acerca do tema sexualidade na terceira idade, abordando assuntos referentes aos preconceitos e tabus nessa fase, além das mudanças decorrentes do envelhecimento e alertas sobre as Infecções Sexualmente Transmissíveis (ISTs). Dessa forma, a dinâmica estabelecida teve como foco principal contribuir para o empoderamento dos idosos, tornando-os capazes de discernir os mitos e estereótipos da sexualidade, bem como instruir quanto aos riscos e as formas de prevenção às ISTs. (AU)

The age pyramid shows progressive ageing in the world. This process involves aspects related to health, including sexuality as an interfering variable in the quality of life of the human being. This report aimed to discuss a health education action directed to senior citizens, carried out within the scope of Primary Health Care, developed by medical students. The experience took place through conversation and counseling concerning the theme 'sexuality in old age', addressing issues related to prejudice and taboos in this phase, as well as changes due to aging and warnings about Sexually Transmitted Infections (STIs). Thus, the established dynamics had as main focus to contribute to the empowerment of the elderly, making them capable of discerning the myths and stereotypes of sexuality, as well as instructing on the risks and ways to prevent STIs. (AU)

Assessment of histological liver alterations in dogs naturally infected with Leishmania infantum.

BACKGROUND: The liver plays a central role in the development of canine visceral leishmaniasis. Studies of natural infection in animals and humans indicate a direct relationship between resolution of infection and the formation and maturation of granulomas in the liver. However, in contrast to other reports in the literature, the present study found no differences in the characteristics of hepatic granulomas that could be related to resistance or susceptibility to Leishmania. Here, we describe the hepatic alterations observed in dogs with differing clinical manifestations of visceral leishmaniasis in an endemic area in the state of Bahia, Brazil. METHODS: We examined 148 animals in an endemic area. The animals were clinically examined, and the infection was determined by ELISA, spleen aspirate culture and quantitative PCR. The animals were grouped into asymptomatic or symptomatic based on the number of signs of LV. The histological liver evaluation was performed in a blinded way. RESULTS: Our results indicated no association between the characteristics of granulomas and clinical presentation. We found an association between the intensity of this inflammatory response and parasite load in the animals' spleens. It is important to note that while hepatic alterations, such as portal and perivascular inflammation and the presence of larger amounts of granulomas, were linked with higher parasite loads, we found the inverse to be true with respect to intrasinusoidal lymphocytosis, the formation of intrasinusoidal inflammatory cell aggregates and Kupffer cell hypertrophy. CONCLUSIONS: Our findings suggest that the presence of mononuclear inflammatory cells inside the sinusoids is more important than that of organized granulomas in terms of the containment of parasitism by the host. We suggest that the presence of granulomas indicates the failure of a first line of defense mechanism in the control of parasite infection, which could be related to the presence of inflammatory cells and Kupffer cell hypertrophy inside the sinusoids. We further demonstrated that dogs with active Leishmania spp. infection present a higher frequency of inflammatory changes in the liver. In addition to being correlated with the severity of clinical manifestation, these hepatic alterations were also associated with changes in hematological and biochemical parameters.

Histological Disorganization of Spleen Compartments and Severe Visceral Leishmaniasis.

The spleen is a secondary lymphoid organ responsible for immune surveillance against blood-circulating pathogens. Absence of the spleen is associated with increased susceptibility to systemic spread and fatal infection by different pathogens. Severe forms of visceral leishmaniasis are associated with disorganization of spleen compartments where cell interactions essential for splenic immunological function take place. White pulp atrophies, secondary lymphoid follicles and marginal zones vanish, and the boundaries separating white and red pulp blur. Leukocyte populations are reduced or disappear or are replaced by plasma cells. In this paper, we review the published data on spleen disorganization in severe forms of visceral leishmaniasis and propose a histological classification to help the exchange of information among research groups.

Histopathological and immunohistochemical characterisation of hepatic granulomas in Leishmania donovani-infected BALB/c mice: a time-course study.

BACKGROUND: Visceral leishmaniasis (VL) is a neglected tropical disease (NTD), caused by the intracellular protozoan parasites Leishmania donovani and Leishmania infantum. Symptomatic VL is considered fatal when left untreated. At present, there is no effective vaccine licensed for human use and available chemotherapies have limitations. Understanding the local immune mechanisms required for the control of infection is a key factor for developing effective vaccines and therapeutics. METHODS: We have investigated the development of the typical granulomatous lesions in the liver in experimental VL over time, together with the local immune responses. BALB/c mice were infected intravenously with a dose of 2 × 107 L. donovani amastigotes (MHOM/ET/67/HU3) and sacrificed at 15, 35 and 63 days post-infection (dpi). Histopathology and immunohistochemical techniques were used for the detection of Leishmania antigen, selected cell types including B and T lymphocytes, macrophages and neutrophils (CD45R-B220+, CD3+, F4/80+ and Ly-6G+) and iNOS. RESULTS: Granulomatous lesions were identified as early as 15 dpi in the livers of all infected animals. Three categories were used to classify liver granulomas (immature, mature and clear). Clear granulomas were exclusively detected from 35 dpi onwards. Kupffer cells (F4/80+) were predominant in immature granulomas, regardless of the dpi. Nonetheless, the highest expression was found 63 dpi. Positive staining for iNOS was mainly observed in the cytoplasm of fused Kupffer cells and the highest expression observed at 35 dpi. T cells (CD3+) and B cells (CD45R-B220+) were predominant in more advanced granuloma stages, probably related to the establishment of acquired immunity. Neutrophils (Ly-6G+) were predominantly observed in mature granulomas with the highest expression at 15 dpi. Neutrophils were lower in numbers compared to other cell types, particularly at later time points. CONCLUSIONS: Our results reflect the role of macrophages during the early stage of infection and the establishment of a lymphocytic response to control the infection in more advanced stages.

Severe clinical presentation of visceral leishmaniasis in naturally infected dogs with disruption of the splenic white pulp.

In this work, we investigated the association between the disruption of splenic lymphoid tissue and the severity of visceral leishmaniasis in dogs. Clinical and laboratory data from 206 dogs were reviewed. Spleen sections collected during the euthanasia of these animals were analyzed, and the splenic lymphoid tissue samples were classified as well organized (spleen type 1), slightly disorganized (spleen type 2), or moderately to extensively disorganized (spleen type 3). Of 199 dogs with evidence of Leishmania infection, 54 (27%) had spleen type 1, 99 (50%) had spleen type 2, and 46 (23%) had spleen type 3. The number of clinical signs associated with visceral leishmaniasis was significantly higher in the animals with evidence of Leishmania infection and spleen type 2 or 3 than in the animals with spleen type 1. Alopecia, anemia, dehydration, dermatitis, lymphadenopathy, and onychogryphosis were all more frequent among animals with evidence of Leishmania infection and spleen type 3 than among the dogs with evidence of Leishmania infection and spleen type 1. The association between the severity of canine visceral leishmaniasis and the disorganization of the splenic lymphoid tissue was even more evident in the group of animals with positive spleen culture. Conjunctivitis and ulceration were also more common in the animals with spleen type 3 than in the animals with spleen type 1. The serum levels (median, interquartile range) of albumin (1.8, 1.4-2.3 g/dL) and creatinine (0.7, 0.4-0.8 mg/dL) were significantly lower and the serum levels of aspartate aminotransferase were significantly higher (57, 39-95 U) in animals with spleen type 3 than in animals with spleen type 1 (2.8, 2.4-3.4 g/dL; 0.9, 0.7-1.2 mg/dL and 23, 20-32 U, respectively). Our data confirm the hypothesis that disruption of the splenic lymphoid tissue is associated with a more severe clinical presentation of canine visceral leishmaniasis.

Three-dimensional systems for in vitro follicular culture: overview of alginate-based matrices.

The in vitro culture of ovarian follicles has provided critical insight into the biology of the follicle and its enclosed oocyte and the physical interaction and communication between the theca and granulosa cells and the oocyte that is necessary to produce meiotically competent oocytes. Various two-dimensional (2D) and three-dimensional (3D) culture systems have been developed to evaluate the effect of growth factors, hormones, extracellular matrix components and culture conditions on follicle development and oocyte growth and maturation. Among these culture systems, 3D systems make it possible to maintain follicle structure and support communication between the various cell compartments within the follicle. In this review article, we will discuss the three main approaches to ovarian follicle culture: 2D attachment systems, 3D floating systems and 3D encapsulated systems. We will specifically emphasise the development of and advances in alginate-based encapsulated systems for in vitro follicle culture.

Effect of chronic administration of tamoxifen and/or estradiol on feeding behavior, palatable food and metabolic parameters in ovariectomized rats.

Tamoxifen (TAM) is a selective estrogen receptor modulator (SERM) used in the treatment of breast cancer; however many women complain of weight gain during TAM treatment. The anorectic effects of estradiol (E) and TAM are well known, although the effects of E on the consumption of palatable food are controversial and there is no information regarding the effects of TAM on palatable food consumption. The aim of this study was to investigate the effects of chronic treatment with estradiol and/or tamoxifen on feeding behavior in ovariectomized rats exposed to standard chow and palatable foods (Froot Loops® or chocolate). Additionally, parameters such as body weight, uterine weight, lipid profile and plasma glucose were also measured. Wistar rats were ovariectomized (OVX) and subsequently injected (ip.) for 40 days with: E, TAM, E+TAM or vehicle (OVX and SHAM - controls). Behavioral tests were initiated 25 days after the start of treatment. Froot Loops® consumption was evaluated in a novel environment for 3 min. Standard chow intake was evaluated for two days and chocolate intake for 7 days in the home cage in a free choice model (chocolate or standard chow). Rats injected with E, TAM and E+TAM groups showed a reduction in body weight and standard chow intake, compared with control groups. With regard to palatable food intake, the E, TAM and E+TAM groups demonstrated increased consumption of Froot Loops®, compared with the SHAM and OVX groups. In contrast, all groups increased their consumption of chocolate, compared with standard chow; however the E group consumed more chocolate than the OVX, TAM and E+TAM groups. Despite these differences in chocolate consumption, all groups showed the same caloric intake during the chocolate exposure period; however the TAM and E+TAM groups presented decreased body weight. Treatment with estradiol and tamoxifen showed a favorable lipid profile with low levels of TC, LDL, LDL/HDL ratio and lower levels of plasma glucose. The E group presented high levels of TG and HDL, when compared with the TAM and E+TAM groups. Taken together, results suggest that TAM acted in an estrogen-like manner on the majority of parameters analyzed. However, tamoxifen acts in a different manner depending on the type of palatable food and the exposure. In addition, the TAM group demonstrated weight loss, compared with other groups independently of the type of food presented (palatable food or standard chow), showing a low caloric efficiency.

Dynamized follicle-stimulating hormone affects the development of ovine preantral follicles cultured in vitro.

OBJECTIVE: To evaluate the effect of dynamized follicle-stimulating hormone (FSH) on the survival, activation and growth of ovine preantral follicles (PFs) in vitro. METHODS: Ovarian fragments were cultured for 1 or 7 days in alpha minimum essential medium (α-MEM(+)) control in the absence or presence of alcohol (Al control) or FSH (6cH, 12cH and 30cH) added at intervals of 24 or 48 h. The ovarian fragments were processed, coded and analyzed by a blinded observer by classical histology (CH), fluorescence microscopy (FM) and transmission electron microscopy (TEM). RESULTS: After 7 days of culture, the group which to which FSH 6cH was added at 24 h intervals showed better rates of follicle survival and activation compared to α-MEM(+) control or Al control (p < 0.05). This group also showed higher follicle and oocyte growth than α-MEM(+) control (p < 0.05). FM and TEM techniques confirmed that FSH 6cH promoted viability and ultrastructural integrity of follicles after 7 days of culture. CONCLUSIONS: FSH 6cH (24 h) treatment maintained the viability, and promoted the activation and in vitro growth of ovine PFs.

Effect of medium composition on the in vitro culture of bovine pre-antral follicles: morphology and viability do not guarantee functionality.

Summary This study investigated the effect of three different culture media (α minimum essential medium (α-MEM), McCoy or TCM199 during the in vitro culture (IVC) of bovine isolated pre-antral follicles. Pre-antral follicles greater than 150 µm in size were isolated and cultured for 0 (control), 8 or 16 days in one of the abovementioned culture media. Follicles were evaluated for survival, growth and antrum formation at days 8 and 16. The results showed that TCM199 was the most suitable medium to preserve follicular viability and ultrastructure, resulting in the highest rates of antrum formation. In conclusion, TCM199 promotes the in vitro development of isolated pre-antral follicles without hampering follicular functionality by sustaining in vitro growth and antrum formation.

Desorganização da polpa branca esplênica está associada com a apresentação clínica mais grave da leishmaniose visceral em cães naturalmente infectados / Disorganization of the splenic white pulp is associated with more severe clinical presentation of visceral leishmaniasis in naturally infected dogs

O baço é o maior órgão linfoide secundário em seres humanos e em cães. Em ambos, a ausência do baço está associada com um risco aumentado de ocorrência de infecções localizadas e disseminadas, incluindo sepse generalizada. A leishmaniose visceral e outras infecções podem alterar a estrutura histológica do baço, o que leva a uma destruição dos microambientes da polpa branca. Trabalhos anteriores mostraram que a uptura da estrutura de polpa branca é mais frequente em cães com marcadores laboratoriais de suscetibilidade à leishmaniose visceral, como a cultura esplênica positiva e LST negativo, do que nos animais em que estes marcadores de susceptibilidade estavam ausentes. Neste estudo, o nosso objetivo é examinar a relação entre a desorganização histológica da polpa branca esplênica e a gravidade da leishmaniose visceral. As amostras e os dados utilizados neste estudo foram coletados de 206 cães de rua provenientes de uma área endêmica para leishmaniose visceral, a cidade de Jequié (Bahia, Brasil). Os animais foram examinados clinicamente e foram realizados os testes ELISA e LST. Aspirados de baço foram coletados para a cultura, e fragmentos de baço foram coletadas para estudos de biologia molecular e de estudos morfológicos. Os animais foram classificados de acordo com o grau de organização estrutural da polpa branca esplênica em grupos com baço (a), bem organizado, (b) ligeiramente desorganizado, (c) a moderadamente a extensivamente desorganizado. Em relação à positividade no ELISA juntamente com a desorganização do baço, conjuntivite ( P= 0,0116), hiperproteinemia (p= 0,021) foram mais frequentes no grupo de animais com ELISA positivo e baço desorganizado, quando comparado com os outros grupos. Os animais polissintomáticos são mais frequentes no grupo com ELISA positivo e baço do tipo 3 (p= 0,004). Os scores clínicos atribuídos à intensidade da conjuntivite (P <0,05), dermatite (P <0,05) e linfadenopatia (P <0,01), alopecia (P <0,01), onicogrifose (P <0,05) foram mais elevados nos animais com ELISA positivo e baço desorganizado do que outros grupos, bem como o número de sinais clínicos atribuíveis à leishmaniose visceral canina (p= 0,0014).Em relação à análise da positividade na cultura esplênica juntamente com a desorganização do baço, a frequência de cães polissintomáticos foi maior em animais com baço ligeiramente desorganizado (P <0,05) e baço desorganizado (P <0,005), do que em animais com baço organizado. Alopecia (P <0,01), conjuntivite (P <0,05), desidratação (P <0,001), dermatite (P <0,05), onicogrifose (p <0,01), anemia (P <0,05), úlcera (P <0,001) e alto escore clínico (P <0,001) foram mais frequentes em animais com cultura esplênica positiva e baço desorganizado, do que nos animais com cultura negativa e baço organizado. Em conclusão, os cães com desorganização do baço associada com leishmaniose visceral têm mais sinais clínicos e pior estado clínico do que os animais com leishmaniose visceral, mas sem desorganização do baço.

BMPRIB and BMPRII mRNA expression levels in goat ovarian follicles and the in vitro effects of BMP-15 on preantral follicle development.

This study evaluated the levels of bone morphogenetic protein receptors BMPRIB and BMPRII mRNA in goat follicles and the effects of bone morphogenetic protein-15 (BMP-15) on the in vitro development of cultured preantral follicles. Real-time polymerase chain reaction (PCR) was used to analyze the levels of BMPRIB and BMPRII mRNA in caprine preantral follicles and in small and large antral follicles. Preantral follicles (≥150 µm) were also isolated from goat ovaries and cultured for 18 days in α-MEM(+) supplemented with or without BMP-15 (10, 50, or 100 ng/ml). At the end of culture, some follicles were fixed for ultrastructural evaluation. Real-time PCR showed a reduction in BMPRII mRNA levels from the primary to secondary follicles. Higher levels of BMPRIB mRNA were observed in granulosa/theca cells from large antral follicles compared with small antral follicles. Moreover, BMPRII mRNA was expressed to a greater extent in cumulus-oocyte complexes from large antral follicles than in their respective granulosa/theca cells. In culture, 50 ng/ml BMP-15 positively influenced antral cavity formation and follicle growth after 18 days and also maintained follicular integrity. Thus, BMPRIB and BMPRII mRNAs are present in all follicular categories. BMP-15 (50 ng/ml) stimulates growth, antrum formation and the ultrastructural integrity of isolated caprine preantral follicles after 18 days of culture.

Ultrastructural features of agouti (Dasyprocta aguti) preantral follicles cryopreserved using dimethyl sulfoxide, ethylene glycol and propanediol.

The objective was to develop an efficient protocol for cryopreservation of agouti (Dasyprocta aguti) ovarian tissue. Agouti ovarian fragments were placed, for 10 min, in a solution containing MEM and fetal bovine serum plus 1.5 M dimethyl sulfoxide (DMSO), ethylene glycol (EG) or propanediol (PROH); some of those fragments were subsequently cryopreserved in a programmable freezer. After exposure and/or thawing, all samples were fixed in Carnoy prior to histological analysis. To evaluate ultrastructure, follicles from the control and all cryopreserved treatments were fixed in Karnovsky and processed for transmission electron microscopy. After exposure and freezing, there was a significant decrease in the percentage of morphologically normal preantral follicles in all treatments when compared to the control (92.67 ± 2.79, mean ± SD). However, there were no significant difference when the exposure and freezing procedures were compared using the same cryoprotectant. Moreover, there was no significant difference among cryoprotectants at the time of exposure (DMSO: 64.7 ± 3.8; EG: 70.7 ± 11.2, PROH: 63.3 ± 8.5) or after freezing (DMSO: 60.6 ± 3.6, EG: 64.0 ± 11.9; PROH: 62.0 ± 6.9). However, only follicles frozen with PROH had normal ultrastructure. In conclusion, preantral follicles enclosed in agouti ovarian tissue were successfully cryopreserved using 1.5 M PROH, with satisfactory maintenance of follicle morphology and ultrastructure.