Microdissection testicular sperm extraction causes spermatogenic alterations in the contralateral testis

Testicular sperm extraction (TESE) associated with intracytoplasmic sperm injection has allowed many men presenting non-obstructive azoospermia to achieve fatherhood. Microdissection TESE (microTESE) was proposed as a method to improve sperm retrieval rates in these patients; however, there have been failures. Little is known about whether microTESE leads to spermatogenic alterations in the contralateral testis. We assessed histological outcomes of experimental microTESE in the contralateral testis of adult male rabbits. Nine adult male rabbits were divided into three groups: control (testicular biopsy to observe normal histological and morphometric values), sham (incision of the tunica vaginalis, and a contralateral testicular biopsy to observe histological and morphometric patterns, 45 days later), and study (left testicular microTESE, and a right testicular biopsy to observe histological and morphometric patterns, 45 days later). Sections were assessed by calculating Johnsen-like scores, and measuring total tubule diameter, lumen diameter and epithelial height. the results were compared using ANOVA and Bonferroni's statistical analysis. Morphometric evaluation of the seminiferous tubules did not demonstrate differences between the three groups. However, microTESE caused spermatogenic alterations, leading to maturation arrest in the contralateral testis.Universidade Federal de São Paulo, Dept Cirurgia, Div Urol, Secao Reprod Humana, São Paulo, BrazilUniv Estadual Paulista, Dept Biol, Inst Biociencias Letras & Ciencias Exatas Sao Jos, Sao Jose Do Rio Preto, SP, BrazilUniversidade Federal de São Paulo, Dept Cirurgia, Div Urol, Secao Reprod Humana, São Paulo, BrazilWeb of Scienc

Caracterização do ciclo nucleolar e da formação do corpo cromatóide na espermatogênese de alguns vertebrados

O corpo cromatóide (CB) é uma organela citoplasmática que, aparentemente, possui um papel no estoque de RNA e proteínas para a diferenciação final dos espermatozóides. Existem algumas teorias que tentam explicar a origem do material que compõe essa organela. Uma dessas teorias, proposta por alguns autores, sugere que o CB se origine a partir de material nucleolar, que se fragmenta nas etapas iniciais da espermatogênese e, em seguida, migra para o citoplasma. O objetivo do presente estudo foi acompanhar o ciclo nucleolar por meio de análises citoquímicas – hematoxilina-eosina (HE); azul de toluidina (AT); variante da concentração crítica de eletrólitos (CEC); reação de Feulgen; impregnação por íons prata (AgNOR); citogenéticas – impregnação por íons prata (AgNOR), e análises ultra-estruturais – microscopia eletrônica de transmissão (MET), para verificar a relação da fragmentação do material nucleolar com a formação do corpo cromatóide (CB), em algumas espécies de vertebrados: Tilapia rendalli (Teleostei, Cichlidae); Dendropsophus minutus (Amphibia, Anura); Phrynops geoffroanus (Reptilia, Testudines) e coelho albino da raça Nova Zelândia – Oryctolagus cuniculus (Mammalia, Lagomorpha). Por meio das análises citoquímicas foi possível observar que ocorre uma fragmentação do material nucleolar no início da prófase I, em todas as espécies analisadas, e uma posterior reorganização do nucléolo no núcleo de espermátides iniciais, com uma área significantemente menor do que a área do nucléolo das espermatogônias. Três fenômenos podem contribuir para essa diferença significante entre as áreas nucleolar de espermatogônias e espermátides: a) Modificação no estado funcional da célula; b) Diminuição no número de RONs nas espermátides; c) Migração de material nucleolar fragmentado...The chromatoid body (CB) is a cytoplasmic organelle that has a function related to RNA and protein accumulation and ⁄ or storage for later germ-cell differentiation. Many theories have been postulated in order to explain the origins of the CB material. One of the most accepted theory describes that it originates from a nucleolar material, where it was fragmented in the early spermatogenesis, and finally, this fragmented nucleolar material migrates to cytoplasm. The aims of the present study were: 1) monitoring the nucleolar material distribution by means of cytochemical techniques (hematoxylin–eosin (HE), toluidine blue (TB), modified Critical Electrolyte Concentration for detecting RNA (CEC), silver-ion impregnation (AgNOR) and Feulgen reaction), and by ultrastructural analysis (Transmission Electron Microscopy – TEM); and 2) comparing the nucleolar material distribution with the formation of CB in some vertebrate species: Tilapia rendalli (Teleostei, Cichlidae); Dendropsophus minutus (Amphibia, Anura); Phrynops geoffroanus (Reptilia, Testudines); and Oryctolagus cuniculus (Mammalia, Lagomorpha). For all analyzed species, the cytochemical techniques showed that the nucleolar fragmentation occurred during the beginning of prophase I, and the nucleolus reorganization occurred in the early spermatids nucleus. Statistical tests evidenced that area of the early spermatids nucleolus were smaller than the spermatogonia nucleolus area. Three phenomena can contribute for the statistical difference between the spermatogonia nucleolar area and the early spermatids nucleolar area: a) Modification of cell activity; b) Decrease of the number of NORs in the spermatids; c) Migration of the fragmented nucleolar material from the nucleus to the cytoplasm. This nucleolar material will participate in the CB formation process. The ultrastructural analysis showed an ...(Complete abstract click electronic access below)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Nucleolar Cycle and Its Correlation with Chromatoid Bodies in the Tilapia rendalli (Teleostei, Cichlidae) Spermatogenesis

The aim of this study was: (1) to monitor the nucleolar material distribution using cytological and cytochemical techniques and ultrastructural analysis; and (2) to compare the nucleolar material distribution with the formation of the chromatoid body (CB) in the germ epithelium of Tilapia rendalli. Nucleolar fragmentation occurred during the leptotene of prophase I and nucleolus reorganization occurred in the early spermatid nucleus. The area of the early spermatid nucleolus was significantly smaller than that of the spermatogonia nucleolus. Ultrastructural analysis showed an accumulation of nuages, which form the CB, before nucleolar fragmentation in the spermatogonia cytoplasm. The CB was observed in association with mitochondrial clusters in the cytoplasm of primary spermatocytes, as well as in those of initial and later spermatids. In conclusion, the nucleolus seems to be related to CB formation during spermatogenesis of T rendalli, because at the moment of nucleolus fragmentation in the primary spermatocytes, the CB reaches its largest area and it is able to complete important functions during spermatogenesis. The reorganized nucleolus of the initial spermatids has a lower area due several factors, one of which is the probable migration of nucleolar fragments from the nucleus to the cytoplasm, therefore playing a role in CB formation. Anat Rec, 293:900-910, 2010. (C) 2010 Wiley-Liss, Inc.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Morphological changes of mammalian nucleoli during spermatogenesis and their possible role in the chromatoid body assembling

Chromatoid body (CB) is a typical cytoplasmic organelle of germ cells, and it seems to be involved in RNA/protein accumulation for later germ-cell differentiation. Despite most of the events in mammals spermatogenesis had been widely described in the past decades and the increase in the studies related to the CB molecular composition and physiology, the origins and functions of this important structure of male germ cells are still unclear. The aims of this study were to describe the nucleolar cycle and also to find some relationship between the nucleolar organization and the CB assembling during the spermatogenesis in mammals. Cytochemical and cytogenetics analysis showed nucleolar fragmentation in post-pachytene spermatocytes and nucleolar reorganization in post-meiotic spermatids. Significant difference in the number and in the size of nucleoli between spermatogonia and round spermatids, as well as differences in the nucleolar position within the nucleus were also observed. Ultrastructural analysis showed the CB assembling in the cytoplasm of primary spermatocytes and the nucleolar fragmentation occurring at the same time. In conclusion our results suggest that the CB may play important roles during the spermatogenesis process in mammals and that its origin may be related to the nucleolar cycle during the meiotic cell cycle

Meiotic nucleolar cycle and chromatoid body formation during the rat (Rattus novergicus) and mouse (Mus musculus) spermiogenesis

The aims of the present study were to follow the nucleolar cycle in spermiogenesis of the laboratory rodents Rattus novergicus and Mus musculus, to verify the relationship between the nucleolar component and chromatoid body (CB) formation and to investigate the function of this cytoplasmic supramolecular structure in spermatogenic haploid cells. Histological sections of adult seminiferous tubules were analyzed cytochemically by light microscopy and ultrastructural procedures by transmission electron microscopy. The results reveal that in early spermatids, the CB was visualized in association with the Golgi cisterns indicating that this structure may participate in the acrosome formation process. In late spermatids, the CB was observed near the axonema, a fact suggesting that this structure may support the formation of the spermatozoon tail. In conclusion, our data showed that there is disintegration of spermatid nucleoli at the beginning of spermatogenesis and a fraction of this nucleolar material migrates to the cytoplasm, where a specific structure is formed, known as the "chromatoid body", which, apparently, participates in some parts of the rodent spermiogenesis process. (c) 2007 Elsevier Ltd. All rights reserved

Efeitos do horário de desempenho de atividade laboral e de hábitos de vida sobre aspectos metabólicos e reprodutivos em uma população da cidade de Bauru-SP - Brasil

Ritmos circadianos podem ser definidos como mudanças em eventos fisiológicos e comportamentais que ocorrem em um período de 24 horas. O objetivo do presente trabalho foi avaliar como mudanças nos ritmos circadianos, exibida por trabalhadores noturnos/de turnos, contribuem para o desenvolvimento e para a prevalência de síndromes metabólicas, como diabetes, hipertensão e obesidade, quando associadas com fatores como idade, gênero e hábitos de alimentação e tabagistas. Esse estudo também considerou a influência de todos os fatores supracitados na manutenção da fertilidade dos indivíduos. Dados foram coletados por meio da aplicação de um questionário respondido por uma população de 600 indivíduos de ambos os gêneros na cidade de Bauru, São Paulo, Brasil. Indivíduos de ambos os gêneros foram agrupados em três categorias de idade: 20-35 anos; 39-59 anos; e acima de 60 anos. Cada categoria de gênero/idade continha 100 indivíduos (n=600; 45 homens e 12 mulheres eram trabalhadores noturnos/turno; e 255 homens e 288 mulheres eram trabalhadores de horário comercial). Nossos resultados não demonstram mudanças nos fatores considerados (incidência de diabetes, hipertensão ou obesidade) entre os trabalhadores noturnos/turno de ambos os sexos mais jovens (20-35 anos). Mulheres trabalhadoras noturnas/turno na categoria “36-59 anos” apresentaram alta incidência de hipertensão, e a hipertensão estava associada com maus hábitos de alimentação e tabagismo. Homens acima de 60 anos que trabalharam em turno/noturno apresentaram alta incidência de diabetes, hipertensão e obesidade e todos esses fatores foram associados com tabagismo. Em conclusão, nossos dados demonstraram o efeito do tempo de exposição às alterações nos ritmos circadianos sobre a prevalência de síndromes metabólicas. Também observamos que mulheres são mais susceptíveis a essas alterações do que homens.  Os dados de fertilidade coletados no presente estudo foram inconclusivos. ABSTRACT Effects of Work Hours and Lifestyle on Metabolic and Reproductive Facets of a Brazilian Population (Bauru, Sao Paulo, Brazil)Circadian rhythms can be defined as changes in physiological and behavioral events within a 24 hours period. The aim of the present study was to evaluate how changes to circadian rhythms in shift workers contribute to the development and the prevalence of metabolic disorders, such as diabetes, hypertension, obesity when associated with factors such as gender, age, and smoking and eating habits. This study also considers the influence of these factors on individual fertility. The study was performed by analyzing data collected in a survey. The survey was answered by a population of 600 individuals of both genders in the city of Bauru, São Paulo State, Brazil. Individuals of both genders were grouped in three different age categories: 20-35 years old; 36-59 years old; and 60 years old and older. Each age/gender category contained one hundred individuals (n= 600; 45 men and 12 women were shift workers, and 255 men and 288 women worked regular business hours). Our results showed no changes in the factors considered (incidence of diabetes, hypertension, and obesity) among younger shift workers (20-35 years old) of either gender in our population. Shift-working women in the “36-59 years old” category presented a high incidence of hypertension, and this hypertension was associated with poor diet and smoking habits. Shift-working men in the “60 years old and older” category presented high incidences of diabetes, hypertension, and obesity; all of these factors were associated with smoking habits. In conclusion, our data showed the effect of the extent of circadian rhythm disruption on the prevalence of metabolic syndrome. We also found that women seem to suffer the effects of the circadian rhythm disruption earlier in life than men. Fertility data were not conclusive in our study

Expression of miRNA-146a, miRNA-155, IL-2, and TNF-alpha in inflammatory response to Helicobacter pylori infection associated with cancer progression

miRNAs appear to play an important role in controlling the expression of several genes, and they are a potential biomarker and prognostic tool in gastric diseases. We analyzed 53 controls, 86 patients with gastritis, and 19 patients with gastric cancer. Real-time-PCR was used to determine the expression levels of miRNA-146a, miRNA-155, IL-2, and TNF-. The subsequent analysis of the target genes was performed using the bioinformatics approach. There was no difference in IL-2 expression between the groups. However, there was a significant increase in TNF- expression in the gastritis group relative to the control and a significant decrease in the gastric cancer group relative to the control. There was also a statistically significant increase in miRNA-146a and miRNA-155 expression in the gastritis group relative to the control, but not in the gastric cancer group. Similar results were found when the presence of H. pylori was considered. The data revealed an increase in miRNA-146a and miRNA-155 expression but not enough to control the expression of TNF-. The presence of H. pylori was found to affect increases in TNF- and microRNA expression, and miRNA-146a and miRNA-155 alone were not able to eliminate bacteria or restore tissue homeostasis.Sao Paulo Research FoundationUniversidade do Sagrado Coração (USC), Bauru, Sao Paulo, BrazilUniv Fed Sao Paulo UNIFESP, Sao Paulo, SP, BrazilUniv Fed Sao Paulo UNIFESP, Sao Paulo, SP, BrazilFAPESP: 2015/11371-5FAPESP: 2015/11613-9Web of Scienc