Localization and hormonal regulation of endometrial matrix metalloproteinase-26 in the rhesus macaque

The current understanding of hormonal regulation of matrix metalloproteinase-26 (MMP-26) in the primate endometrium is incomplete. The goal of this work was to clarify estrogen and progesterone regulation of MMP-26 in the endometrium of ovariectomized, hormone-treated rhesus macaques.Ovariectomized rhesus macaques (n 66) were treated with estradiol (E-2), E-2 plus progesterone, E-2 followed by progesterone alone or no hormone. Endometrium was collected from the hormone-treated animals during the early, mid- and late proliferative and secretory phases of the artificial menstrual cycle. MMP-26 expression was quantified by real-time PCR, and MMP-26 transcript and protein were localized by in situ hybridization and immunohistochemistry and correlated with estrogen receptor 1 and progesterone receptor (PGR).MMP-26 was localized to glandular epithelium and was undetectable in the endometrial stroma and vasculature. MMP-26 transcript levels were minimal in the hormone-deprived macaques and treatment with E-2 alone did not affect MMP-26 levels. Treatment with progesterone both in the presence and absence of E-2 stimulated MMP-26 expression in the early and mid-secretory phases (P 0.001). MMP-26 expression preceded decidualization of endometrial stroma. MMP-26 levels then declined to baseline in the late secretory phase (P 0.01) despite continued E-2 plus progesterone treatment. Loss of detectable MMP-26 expression in the late secretory phase was correlated with late secretory phase loss of glandular epithelial PGR.Endometrial MMP-26 expression is dependent on the presence of progesterone in the early secretory phase and then gradually becomes refractory to progesterone stimulation in the late secretory phase. In the macaque, MMP-26 is a marker of the pre-decidual, secretory endometrium. During the second half of the late secretory phase, and during decidualization, MMP-26 loses its response to progesterone concurrent with the loss of epithelial PGR. The decline in MMP-26 levels between the mid- and late secretory phases may play a role in the receptive window for embryo implantation.

Caracterização da fusão caruncular em gestações naturais e de conceptos bovinos clonados

Barreto R.S.N., Miglino M.A., Meirelles F.V., Visintin J.A., Silva S.M., Burioli K.C., Fonseca R., Bertan C., Assis Neto A.C. & Pereira F.T.V. 2009. [Characterization of the caruncular fusion in gestations of natural and cloned bovine conceptuses.] Caracterizacao da fusao caruncular ern gestacoes naturais e de conceptos bovinos clonados. Pesquisa Veterinaria Brasileira 29(10):779-787. Laboratorio de Morfofisiologia da Placenta e Embriao, Faculdade de Zootecnia, Universidade Estadual Paulista, SP294 Km 651, Dracena, SP 17900-000, Brazil. E-mail: fverechia@dracena.unesp.brThe objective of the study was to compare the characteristics of the caruncular fusion in gestations of non-cloned and cloned conceptuses. The non-cloned conceptuses were divided according to the gestation period: Group 1 (2 to 3 months; n=9),II (4 to 6; n=9); III (7 to 8; n=10) and IV (9 n=7). The cloned conceptuses formed the Group V: 9 months; n=4. The caruncles were observed macroscopically (number and dimensions: length, width and height), microscopically and submitted to statistical analysis (5% of significance). We observed three types of macroscopic caruncular fusions: oval (morphologically normal); two united adjacent caruncles and the lobulated type, characterized by regions with several united caruncles presenting a false fusion or deformation of the caruncular parenchyma. The length of the caruncles was 1.55 +/- 0.57; 2.45 +/- 0.55; 4.66 +/- 2.0 and 5.72 +/- 1.90cm for the groups 1, 11, 111, IV respectively. As for the height, the caruncles presented a lineal growth during the gestation: 0.40 +/- 0.15; 0.57 +/- 0.21; 1.0 +/- 0.48 and 1.80 +/- 0.91cm, for the respective groups 1, 11, 111 and IV. The width of the caruncles was similar between the groups I and 11 (0.97 +/- 0.30 e 1.42 +/- 0.71 cm) and the groups III and IV (2.68 +/- 1.22 and 3.52 +/- 1.16cm). When the group V was compared to the IV, the caruncles of the group V presented a larger length (5.72 +/- 1.90 vs. 7.88 +/-.13cm) and width (3.52 +/- 1.16 vs. 4.93 +/- 1.46cm), however they were similar in height (1.80 +/- 0.91 and 2.25 +/- 0.67cm). We verified that in gestations of cloned conceptuses the caruncles presented a larger development than in gestations of non-cloned conceptuses. The fusioned caruncles presented measurements statistically similar to the isolated ones in all the parameters and groups. Under light microscopy, we observed the formation of a stromal axis from the basis of the caruncle to the apex of the fusional fissure, with the histological constitution similar to the endometrial stroma. Three microscopic shapes were also unpublished defined: true fusion with a single axis evident below the fusional fissure; pseudofusion with a double axis in H shape and false fusion with absence of the axis. The first two formats were associated to the oval and lobulated caruncles and the last one to the false fusion with deformation of the caruncle parenchyma. The fusional axis increased in size along the gestation among the groups I, II, III and IV. The group V presented a larger length and width of the axis when compared to the group IV. Thus, in gestations of cloned conceptuses a destruction of the lateral epithelium of the caruncles is associated to an incompetence in the maternal-fetal interdigitation, that compromises the cotyledonary fusion. We suggest that, in gestations derived of cloned conceptuses, the increase of the size of the caruncular fusions is possibly associated to a compensatory mechanism for the metabolic exchanges between mother and fetus, in reason of the smallest number of isolated caruncles.

Morphological and Histochemical Characterization of Gill Filaments of the Brazilian Endemic Bivalve Diplodon expansus (Kuster, 1856) (Mollusca, Bivalvia, Hyriidae)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

TROMBOSE VENOSA EXPERIMENTAL NA COBAIA: EFEITO DA HEPARINA E DE DROGAS QUE ALTERAM A FUNCAO PLAQUETARIA

Thrombosis was induced in the jugular vein of guinea pigs by damage of the epithelium with a sclerosing solution. The effect of heparin, Dextran 70, acetylsalicylic acid, dipyridamole and phenylbutazone in preventing the development of the thrombus was investigated. Only heparin in high doses showed this effect.

Fine structure of lamellated nerve endings in the gingiva of man and the Cebus apella monkey

Gingival mucosae of man and the adult Cebus apella monkey were fixed for 3 hr in modified Karnovsky fixative containing 2.5% glutaraldehyde, 2% formaldehyde in 0.1 M sodium phosphate buffer (pH=7.4). The specimens were postfixed in 1% osmium tetroxide in 0.1 M sodium phosphate buffer at 4°C for 2 hr, dehydrated in a graded alcohol series and embedded in Epon 812. Thick sections of 1-3 μm and ultrathin sections of 40-80 nm in thickness were cut with glass knives on an LKB ultramicrotome. The thick sections were stained with toluidine blue solution, and the grids were stained with uranyl acetate and lead citrate and examined under a Philips EM-301 electron microscope. Our observations permitted us to conclude that: both gingival mucosae, of man and the Cebus apella monkey, have lamellar nerve endings; these corpuscles are localized in the papillar space of the epithelium and do not contact closely with the basement membrane; the nerve endings are composed of an afferent fiber which subdivides several times and forms irregular flattened or discoidal expansions; the laminae of the lamellar cells are very thin near the terminal axon and are larger and irregular in shape at the peripheral portion of the corpuscle; the terminal axon shows abundant mitochondria, myelin figures, clear vesicles, and multivesicular bodies; between the axoplasm membrane and adjacent cytoplasmic lamina and between the lamellae, small desmosome type junctions are noted; and the cytoplasmic material of the lamellae cells is characterized by the presence of numerous microfilaments, microtubules, mitochondria, rough endoplasmic reticulum, and caveolae.

Urinary bladder washings in autopsies. A cytopathologic study of 63 cases

The sediment from urinary bladder washings from 63 consecutive autopsies was cytologically studied in order to achieve a better understanding of the changes in urothelial cells collected from hospital populations. The observed alterations were correlated with alterations in the urinary system and with therapy preceding death. The specimens obtained were of good quality. In 39.7% of the cases, the sediment contained giant superficial multinucleated cells. Three of nine cases previously subjected to radiation or chemotherapy showed atypical urothelial cells. In three cases with immunosuppression, there was cytologic evidence of subclinical infection by polyomavirus, and virus particles were identified by electron microscopy of the vesical mucosa. The study of the smear background offered additional information: the sediment contained hyaline or hematic or hyaline-cellular casts in 17.4% of the cases, in all of which there were renal tubulopathies when the kidney sections were studied. The method is useful for a good evaluation of the autopsy as well as for training in urinary cytopathology.

Características histoquímicas e estruturais do testículo do morcego hematófago (Desmodus rotundus rotundus, Geoffrey, 1810).

The testicular stroma of the vampire bat including the testicular capsula and the lamina propria of the seminiferous tubuli, was strongly PAS-positive. This observation was a possible indication of great amounts of structural glycogen and other glycoconjugates at the level of smooth muscle cells; elongated contractile cells and/or collagen frameworks of the tunica albuginea and tubular lamina propria. In the last the basement membranes of the seminiferous tubules were particularly strongly PAS positive, as an indication of their neutral mucosubstances structural composition, previously described (Malmi et al., 1987). The epithelium lining from the cavitary and surface rete testis complex showed low reactivities to mucosubstances; total proteins and lipids and oxidative enzymes studied. Although the apical granulation at the rete testis epithelium showed an intense PAS reactivity with hypothesis of glycoprotein secretion, through the rete. The PAS, Sudan Black B, NADH, MDH and LDH reactions of the testicular interstitium seem correlate to steroid metabolism (biosynthesis and secretion), at the Leydig cells level. The seminiferous epithelium generally had low reactions to all the histochemical studies realized. Particularly in the adbasal compartment the histochemical localizations of NADH diaphorase and LDH were possible related to glycolytic activities and general carbohydrates metabolism, both enzymes, and hydrogen transport, the NADH. The strong PAS, diastase and PAS, and alcian blue pH 2.5 and PAS reactions observed in the adluminal seminiferous epithelium compartment were directly related to the spermatids acrosomal glycoconjugates structuration. Also the SDH localization at this level seems to be related to the mitochondrial activities at the middle piece level in the late spermatids.

Fine structure of the ductuli efferentes of the hamster (Mesocricetus auratus).

Structurally the ductuli efferentes of the hamster showed 2 distinct segments, a testicular and an epididymal. Both of these segments were lined by a pseudostratified epithelium, which showed basically non-ciliated and ciliated cells. In the testicular segment a 3rd type of oval dark cells was observed. The ultrastructural characteristics of these cells were presented and discussed in this report.

Influence of different types of non-resorbable suture material on the healing of extraction wounds--a histological study in rats.

The influence of different non-resorbable suture materials on the healing of tooth extraction wounds was examined in 120 adult male albino rats. Nylon gave the best biological results for healing of the mucosa and dental socket. Silk and cotton produced considerable delay of socket healing. Healing was only mildly delayed by polyester suture, with a short moderate inflammatory reaction at the initial stage.

Estudo histológico comparativo da mucosa palatina em pacientes portadores e não portadores de próteses totais antes e após a instalação de novas proteses.

The authors verified histologically in 20 no dentates patients, the influence of complete dentures on the hard palate mucosa. The patients were divided in two groups: I. Patients that never used complete dentures; II. Patients that were using complete dentures when this research was realized. New prosthesis were made for each one of the 20 patients. Immediately before they received the new prosthesis and ninety days after this, biopsies were realized. The circunferencial pieces of the hard palate mucosa were fixed in formol 10% and coloured with Hematoxylin/Eosin and Mallory's Tricomic for histomorphologic analysis and measuring of the stratum corneum of the epithelium. It was made the statistical analysis and the results showed that: 1. In normal conditions, the utilization of the full dentures produces the reduce of the thickness of the stratum corneum. This was more observed in Group I. 2. The range difference of the thickness of the stratum corneum between the first and the second biopsies in the groups I and II (4.86 microns and 2.18 microns) were not statistically significant. 3. With the use of complete dentures the stratum corneum of the epithelium showed tendence to parakeratinization.

Características ultra-estruturais das células de Sertoli do morecego hematófago (Desmodus rotundus rotundus, Geoffrey, 1810).

This paper deals with the ultrastructural study of mature vampire bat Sertoli cells and their relationships with the different stages of testicular germ cells. In vampire bat seminiferous epithelium there are different types of junctional specializations among Sertoli cells and among Sertoli cells and different germ cells, with special emphasis to tight junctions and to junctions like as desmosomes. Ectoplasmic junctions through the Sertoli cells, including the smooth ER, are observed. These cellular interactions and their cytophysiological roles are discussed. Also are related some ultrastructural peculiarities of the Sertoli cell nucleus, nucleolus, cytoplasmic organelles and lipidic inclusions.

Infection and apparent invasion of Vero cells by Paracoccidioides brasiliensis

Paracoccidioides brasiliensis probably uses many different mechanisms to establish itself in the host and cause disease. In this work, we assess an in vitro model system which uses cultured mammalian cells to investigate the virulence factors of P. brasiliensis. We were able to demonstrate an invasion process of the yeast form of this fungus in Vero cell cultures. We deduced that the overall invasive process involved three steps: adhesion, followed by invasion of individual epithelial cells and spread to adjacent cells.

Gut intraepithelial lymphocyte counts in neonates, infants and children

Intraepithelial lymphocyte (IEL) counts were histologically assessed in the jejunum, ileum and appendix of 39 neonates (0-28 days), 32 infants (1-9 months) and 13 children (1-9 years). Small intestinal mucosa samples were obtained from 73 autopsies, and from 8 surgical and 3 aspirative biopsies. IEL counts of specimens from the jejunum, ileum and appendix gave similar results in the same patient. The number of IEL counts was significantly lower in neonates for all three segments. The difference between infants and children was more marked in the jejunum than in the ileum, although this was not significant. In the appendix, there was no difference between the different age groups. Our results indicate that postnatal expansion of IEL occurs homogeneously along the gut after the neonatal period.

Simple mucin-type carbohydrate antigens (T, sialosyl T, Tn and sialosyl-Tn) in breast carcinogenesis

Immunohistochemical analysis of the expression of simple mucin-type carbohydrate antigens (Tn, sialyl-Tn and T) was performed in a series of 43 cases of intraductal hyperplasia without atypia, 9 cases of intraductal hyperplasia with atypia, 54 cases of ductal carcinoma in situ (DCIS) and 26 cases of invasive breast carcinoma. We also studied 36 cases of isolated breast normal epithelium, 20 cases of 'normal' breast epithelium adjacent to neoplasms and 14 cases of apocrine metaplasia. All antigens were detected in different frequencies in normal, hyperplastic, metaplastic and neoplastic breast epithelium. Tn and sialyl-Tn are expressed more frequently in malignant than in benign breast epithelium; while Tn expression increases from normal to invasive carcinomas, sialyl-Tn increases until DCIS and drops in invasive carcinomas, suggesting that either there is a failure of a proportion of DCIS to progress to invasive carcinoma or loss of expression of sialyl-Tn when some carcinomas become invasive. The high frequency of Tn and sialyl-Tn expression in breast intraductal proliferations probably reflects incomplete glycosylation in these lesions, which is a well-known tumour-associated phenomenon and supports the assumption that such lesions are putative precursors of breast cancer. T antigen was expressed in all groups studied, but its prevalence differed significantly between normal and neoplastic epithelium. The expression of these antigens in epithelium adjacent to carcinomas is similar to that found in isolated normal breast epithelium, whereas apocrine metaplasia has a pattern of simple mucin-type glycosylation that is specific and distinct from that of the normal breast epithelium, with a high frequency of marked expression of Tn and sialyl-Tn. The similarity of the pattern of expression of simple mucin-type antigens in metaplasia and malignant neoplasia reduces the usefulness of these markers from a diagnostic standpoint.