Does orchidopexy revert the histological alterations in epididymal and vas deferens caused by cryptorchidism?

Cryptorchidism is a pathological condition in which the testicles are retained in the abdominal cavity, resulting in atrophic seminiferous tubules. Some gross structural ab-normalities and histological alterations have been described in the epididymis and vas deferens in, humans with cryptorchidic testes. Orchidopexy surgery restores testicular spermatogenesis in experimental and clinical procedures, but it is still unclear whether histological changes in the epididymis and vas deferens caused by cryptorchidism may be reverted by orchidopexy. The aim of this study was to evaluate the histological changes in the epididymis and vas deferens following experimental uni- and bilateral cryptorchidism in mature and immature mice, and to determine whether alterations could be reversed by orchidopexy. Young and adult C57 BL6 mice were randomized into three groups: control mice, bi/unilaterally cryptorchidic mice and bilaterally cryptorchidic mice with orchidopexy. After evaluation of testis, epididymis and vas deferens, there were no histological alterations in contralateral epididymis of mice unilaterally cryptorchidic. Ipsilateral epididymis of unilaterally cryptorchidic mice and epididymis from bilaterally cryptorchidic price showed significant histological alterations. Orchuidopexy restored normal spermatogenesis and the histological features of epididymis. It would appear that persistent male infertility clinically observed after orchidopexy could not be related to histological alteration, in the testis and epididymis. Development and maintenance of the vas deferens seems to be controlled independently of the epididymis since it was not altered by cryptorchidism condition.51210911

The effect of age on the structure and composition of rat tendon fibrocartilage

Biochemical and morphological aspects of fibrocartilages of calcaneal and deep digital flexor tendons in rats aged 30, 180 and 730 days were analyzed. In both tendons a stronger staining with Alcian blue, indicating the presence of proteoglycans, was detected in rats of 30 and 180 days. In animals 730 days old, it was restricted to the pericellular area. Ultrastructural analysis showed a more prominent pericellular matrix in calcaneal tendon compared to the deep digital flexor tendon. The biochemical analysis showed higher levels of proteins and glycosaminoglycans in the calcaneal tendon of 30-day-old rats compared to older rats. In the deep digital flexor tendon, no significant differences were observed between ages. The small proteoglycan, fibromodulin, was detected in both tendons of all ages, but in young rats it appeared to be running as a 210 kDa component, probably due to the association with collagen chains or self-association. (c) 2006 International Federation for Cell Biology. Published by Elsevier Ltd. All rights reserved.31657057

Subset classification of mouse uterine natural killer cells by DBA lectin reactivity

Uterine Natural Killer (uNK) cells are a transient lymphocyte population found in the pregnant uteri of human and rodents. The pregnant uterine environment appears to influence migration, differentiation and suppression of the cytolytic activation of uNK cells but the mechanisms involved in these processes are not well understood. Similarities to circulating NK (cNK) cells are limited. The present study sought to discrimate uNK cells from cNK cells in mice by identification of a unique uNK cell marker. Dolichos biflorus (DBA) lectin, which has high selectivity for glycoconjugates containing N-acetyl D-galactosomine in the terminal position, reacted with the plasma membranes of mouse uNK cells. DBA lectin did not react with other uterine lymphocytes or with cNK cell surfaces in Swiss, CBA-J, C57BL/6, SJL, BALB/c, DBA-2 mice strains. DBA lectin staining was useful for both light and electron microscopy and distinguished 4 uNK cell subtypes that appear to be stages of differentiation. Quantitative evaluation of these 4 uNK cell subtypes over early to late gestational times showed dynamic changes between immature and mature forms in different compartments of the implantation sites and indicated the occurrence of microdomains in the uterus capable of controlling uNK cell proliferation and differentiation. This is the first report showing mouse uNK cells expressing specific molecules not found in other NK cells. Use of this reagent should enhance studies of earlier, non-granulated forms of uNK cells and provide new strategies for purification of mouse uNK cells for functional and molecular studies. (C) 2003 Elsevier Science Ltd. All rights reserved.24547948

Subdermal implants of poly(L-lactic acid) with plasticizer: an ultrastructural study in rats

Poly(L-lactic acid) (PLLA) membranes containing 7% triethylcitrate plasticizer were implanted in the subcutaneous tissue of rats, and the cellular reaction was evaluated over a period of 2-180 days. The samples were processed for conventional transmission electron microscopy. Polymorphonuclear-type cells and a fibrin network were seen within membrane pores 2 days after implantation. In subsequent samples, there was cellular infiltration, which consisted mainly of fibroblasts, macrophages and multinuclear giant cells embedded in an abundant extracellular matrix containing a network of collagen fibers and blood vessels. At 90 and 180 days after implantation, a high density of voluminous phagocytic cells with a large number of endocytic polymer fragments within their cytoplasm was seen. These results show that PLLA membranes can support connective tissue proliferation and remodeling, which are important properties for successful bio-protheses.174167117718

Control of intracellular movement of connexins by E-cadherin in murine skin papilloma cells

The gap junctional intercellular communication-deficient mouse skin papilloma cell line P3/22 expresses Cx43 but not E-cadherin. The E-cadherin gene-transfected cells (P3E1) communicate ina calcium-dependent manner and they were used to study how E-cadherin restores the function of connexins. At low calcium, Cx43 molecules remain in the cytoplasm of P3E1 cells and appear at cell-cell contact areas only in high-calcium medium. While Cx43 is unphosphorylated in P3E1 cells in low-calcium medium, two phosphorylated bands appeared at high calcium. However, when Cx26, which has no C-terminal tail that can undergo phosphorylation, was expressed in P3E1 cells, this connexin also moved to the plasma membrane after the calcium shift and partly colocalized with Cx43, suggesting that C-terminal phosphorylation is not essential for E-cadherin-mediated intracellular transport of connexins. In low calcium, both Cx26 and Cx43 remained and colocalized in the endoplasmic reticulum. As early as 30 min after the shift to high-calcium medium, both Cx43 and Cx26 began to accumulate in the Golgi apparatus. Intracellular movement of connexins to the cytoplasmic membrane at high calcium was effectively blocked by cytochalasin D and brefeldin A. These results suggest that E-cadherin junction formation at high calcium leads to formation of actin cables, which directly or indirectly transport connexins from the cytoplasm to the cell-cell contact membranes via the Golgi apparatus. (C) 2001 Academic Press.270223524

Increased response of Vero cells to PHBV matrices treated by plasma

The copolymers poly(3-hydroxybutyric acid-co-3-hydroxyvaleric acid) (PHBV) are being intensely studied as a tissue engineering substrate. It is known that poly 3-hydroxybutyric acids (PHBs) and their copolymers are quite hydrophobic polyesters. Plasma-surface modification is an effective and economical surface treatment technique for many materials and of growing interest in biomedical engineering. In this study we investigate the advantages of oxygen and nitrogen plasma treatment to modify the PHBV surface to enable the acceleration of Vero cell adhesion and proliferation. PHBV was dissolved in methylene chloride at room temperature. The PHBV membranes were modified by oxygen or nitrogen-plasma treatments using a plasma generator. The membranes were sterilized by UV irradiation for 30 min and placed in 96-well plates. Vero cells were seeded onto the membranes and their proliferation onto the matrices was also determined by cytotoxicity and cell adhesion assay. After 2, 24, 48 and 120 h of incubation, growth of fibroblasts on matrices was observed by scanning electron microscopy (SEM). The analyses of the membranes indicated that the plasma treatment decreased the contact angle and increased the surface roughness; it also changed surface morphology, and consequently, enhanced the hydrophilic behavior of PHBV polymers. SEM analysis of Vero cells adhered to PHBV treated by plasma showed that the modified surface had allowed better cell attachment, spreading and growth than the untreated membrane. This combination of surface treatment and polymer chemistry is a valuable guide to prepare an appropriate surface for tissue engineering application.19263564

In vivo and in vitro Leishmania amazonensis infection induces autophagy in macrophages

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Autophagy is the primary mechanism of degradation of cellular proteins and at least two functions can be attributed to this biological phenomenon: increased nutrient supply via recycling of the products of autophagy under nutrient starvation; and antimicrobial response involved in the innate immune system. Many microorganisms induce host cell autophagy and it has been proposed as a pathway by which parasites compete with the host cell for limited resources. In this report we provide evidence that the intracellular parasite Leishmania amazonensis induces autophagy in macrophages. Using western blotting, the LC3II protein, a marker of autophagosomes, was detected in cell cultures with a high infection index. Macrophages infected with L. amazonensis were examined by transmission electronic microscopy, which revealed enlarged myelin-like structures typical late autophagosome and autolysosome. Other evidence indicating autophagy was Lysotracker red dye uptake by the macrophages. Autophagy also occurs in the leishmaniasis skin lesions of BALB/c mice, detected by immunohistochemistry with anti-LC3II antibody. In this study, autophagy inhibitor 3-methyladenine (3MA) reduced the infection index, while autophagy inductors, such as rapamycin or starvation, did not alter the infection index in cultivated macrophages, suggesting that one aspect of the role of autophagy could be the provision of nutritive support to the parasite. (c) 2012 Elsevier Ltd. All rights reserved.446401408Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Immunocytochemical Studies of Adhesion Molecules on Mouse UNK Cells and Their Extracellular Matrix Ligands During Mouse Pregnancy

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Uterine natural killer (uNK) cells are the dominant lymphocytes of pregnant mammals' uterus Studies have identified four differentiation stage of mouse uNK cells based on Dolichos biflorus lectin cytochemistry, and their distribution showed preferential domain in the uterus through out the pregnancy. This work was done to investigate the expression of alpha 5, alpha 6, and beta 7 integrins on uNK cells and their ligands distribution. Section of mouse uterus from sixth to seventeenth gestational days were submitted to immunocytochemistry and positive reactions for alpha 5, alpha 6, and beta 7 integrins were found on uNK from eighth to tenth gestational days but not after twelfth gestational days. Fibronectin reactions were seemed from sixth to tenth gestational days around uNK from the myometrium and endometrium close to the myometrium No reaction for fibronectin was seen in the decidualized and nondecidualized endometrium near the placenta Laminin reaction was seen Just in the antimesometrial side beta 7 integrin seems to be the active receptor to bind with VCAM-1. or MAdCA.M-1 of endothelial cells, promoting the uNK cross through the vessels. The absence of laminin in an uNK domain suggests these cells are not dependent of laminin and alpha 6 integrin for their establishment. However, fibronectin seems to support uNK migration, proliferation, differentiation, and survival in the uterus by binding with alpha 5 integrin. The loss of alpha 5 integrin ligation by the clown regulation or fibronectin could inhibits these events and further studies are need to investigate whether unligated alpha 5 can actively and initiate apoptosis, maybe in a caspase 8-dependent way that has been called integrin-mediated death. Anat Rec, 293 1081-1088,2010 (C) 2010 Wiley-Liss, Inc293610811088Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq

Induction of apoptosis in HT29 human intestinal epithelial cells by the cytotoxic enterotoxin of Aeromonas hydrophila

The cytotoxic enterotoxin produced by Aeromonas hydrophila is considered to be the main virulence factor in gastrointestinal infections mediated by this pathogen. In this study, we examined the morphological and apoptotic effects of this toxin on HT29 cells, using light and electron microscopy in situ, as well as agarose gel electrophoresis of cell DNA. Cells treated with the cytotoxic enterotoxin became round and lost their polarity as well as their adhesion to each other and to the substrate. Cytoplasmic blebbing and nuclear condensation also occurred. DNA fragmentation was detected by TUNEL labelling and agarose gel electrophoresis. These results show that the cytotoxic enterotoxin of A. hydrophila can induce apoptosis in human intestinal cells in culture.79452553