Caracterização morfológica do útero e tubas uterinas de fetos bovinos da raça Nelore (Bos primigenius indicus) em diferentes fases de gestação

Para a realização desta pesquisa, foram coletados vinte fetos fêmeas em diferentes fases de gestação da raça Nelore. Os cornos uterinos e as tubas uterinas foram dissecados, mensurados e os fragmentos fixados em paraformaldeído tamponado a 4,00%, processados e incluídos em paraplástico. Os cortes com 5 mm foram corados com hematoxilina e eosina, tricrômio de Masson (para fibras colágenas), Verhoeff (para fibras elásticas) e com reticulina (para fibras reticulares). Os resultados mostraram que não há diferença significativa das mensurações entre os lados direito e esquerdo para os cornos uterinos e tubas uterinas, porém há correlação entre os valores obtidos das mensurações dos órgãos em função da idade dos fetos, ou seja, o crescimento dos órgãos acompanha o crescimento fetal. O epitélio de revestimento do útero não apresenta variações morfológicas evidentes no período analisado. A partir de 23 semanas de gestação, a mucosa uterina apresenta evolução marcante no desenvolvimento das projeções e não há aparecimento de glândulas endometriais na parede uterina no período analisado. A camada muscular apresenta a subcamada circular interna desenvolvida até 23 semanas de gestação e a partir de 24 semanas há presença das duas subcamadas. A camada serosa é típica e não mostra variabilidade no decorrer da gestação. As tubas uterinas apresentam diferenças de crescimento, principalmente dos pregueamentos que a partir de 23 semanas de gestação tornaram-se mais altos e ramificados, porém sem aparecimento de pregas terciárias. Com o desenvolvimento fetal, os cílios epiteliais do pregueamento tubárico são maiores em tamanho e em número. Até 32 semanas de gestação, a camada muscular das tubas uterinas apresenta apenas a subcamada circular interna. A camada serosa e o mesosalpinge são típicos e não apresentam variações importantes. As variações mais marcantes para os órgãos estudados ocorreram a partir de 23 semanas de gestação.

Efficacy and safety of a soy isoflavone extract in postmenopausal women: A randomized, double-blind, and placebo-controlled study

Objective: To investigate the efficacy of soy isoflavone on climacteric symptoms in postmenopausal women.Design: In this double-blind, randomized, placebo-controlled study, a total of 80 women (mean age =55.1 years), who reported 5 or more hot flush episodes per day, were randomized to receive either 250 mg of standardized soy extract (Glycine max AT) a total of 100 mg/day of isoflavone (n=40) or placebo (n=40). Exclusion criteria included: contra-indication for hormone therapy (HT), chronic gastrointestinal diseases, and users of HT within the preceding 6-months. For 10-months, climacteric symptoms were evaluated using a score card and the menopausal Kupperman index. Compliance and safety were also assessed. At baseline and the end of the study, lipid and hormonal profiles, as well as vaginal, mammographic and ultrasonographic parameters were measured. The t-test, Wilcoxon test and ANOVA were used in the statistical analysis.Results: At baseline, the mean number of hot flushes was 9.6 +/- 3.9 per day in the isoflavone group and 10.1 +/- 4.9 in the placebo group (p>0.05). After 10 months, there was a significant reduction in frequency of hot flushes among isoflavone users when compared to those on placebo (3.1 +/- 2.3 and 5.9 +/- 4.3, respectively) (p<0.001). Kupperman index mean values showed a significant reduction in both groups. However, soy isoflavone was significantly superior to placebo, in reducing hot flush severity (69.9% and 33.7%, respectively) (p<0.001). Endometrial thickness, mammography, vaginal cytology, lipids and hormonal profile did not change in both groups. No serious adverse event related to isoflavone treatment was reported.Conclusions: the soy isoflavone extract exerted favorable effects on vasomotor symptoms and good compliance, providing a safe and effective alternative therapeutic for postmenopausal women. (C) 2007 Elsevier B.V.. All rights reserved.

THE LONG-TERM EVOLUTION OF IMMUNE DEPOSITS IN PASSIVE HEYMANN NEPHRITIS

1. To study the long term course of passive Heymann nephritis (PHN), 42 adult male Wistar rats were injected with rabbit anti-FX1A serum (PHN group) and 42 rats received normal rabbit serum (control group). Two animals from each group were sacrificed 2 weeks after the inoculation and 10 animals each from the control and PHN groups were sacrificed 4, 13, 25 and 53 weeks later.2. The PHN group exhibited a significant elevation in 20-h proteinuria which lasted from the first week (control group, 9.19 +/- 0.87; PHN group, 25.3 +/- 2.66) to the 25th week (control group, 22.6 +/- 2.15; PHN group, 66.7 +/- 10.4) except for week 17. From week 29 to week 53 there was no statistical difference between the 2 groups.3. Light microscopy showed no difference between the kidneys of PHN and control rats. Immunofluorescence microscopy in PHN rats showed granular deposition of autologous and heterologous IgG on the glomerular basement membrane (GBM), whose intensity and pattern did not change during 53 weeks of observation.4. When examined by electron microscopy the glomeruli of PHN rats showed: a) electron-dense deposits which were initially subepithelial and homogeneous and later intramembranous, granular and often surrounded by an electron-transparent halo; b) focal thickening of the GBM at the sites of intramembranous deposits; c) effacement of podocytes located close to the deposits; d) penetration of the podocytes into the GBM associated with the deposits; e) presence of osmiophilic granules in the cytoplasm of the podocyte located inside the GBM similar to the granules of the deposits next to them. The association of the penetration of the podocytes into the GBM with the deposits and the presence of the osmiophilic granules inside the foot process have not been described previously in PHN.5. The results suggest that the podocytes play a role in the clearing of intramembranous deposits in PHN.

Bone regeneration at implants placed into extraction sockets of maxillary incisors in dogs

AimTo compare the influence of autologous or deproteinized bovine bone mineral as grafting material on healing of buccal dehiscence defects at implants installed immediately into the maxillary second incisor extraction socket in dogs.Material and methodsIn the maxillary second incisor sockets of 12 Labrador dogs, implants were installed immediately following tooth extraction. A standardized buccal defect was created and autologous bone particles or deproteinized bovine bone mineral were used to fill the defects. A collagen membrane was placed to cover the graft material, and the flaps were sutured to fully submerge the experimental areas. Six animals were sacrificed after 2 months, and six after 4 months of healing. Ground sections were obtained for histological evaluation.ResultsAfter 2 months of healing, all implants were osseointegrated. All buccal dehiscence defects were completely filled after 2 months irrespective of the augmentation material (autologous bone or Bio-Oss (R)) applied. Bone-to-implant contact (BIC) on the denuded implant surfaces was within a normal range of 30-40%. However, the newly formed tissue at 2 months was partially resorbed (> 50% of the area measurements) after 4 months.ConclusionsApplying either autologous bone or deproteinized bovine bone mineral to dehiscences at implants installed immediately into extraction sockets resulted in high degree of regeneration of the defects with satisfactory BIC on the denuded implant surface.To cite this article:De Santis E, Botticelli D, Pantani F, Pereira FP, Beolchini M, Lang NP. Bone regeneration at implants placed into extraction sockets of maxillary incisors in dogs.Clin. Oral Impl. Res. 22, 2011; 430-437.

Mummified papyraceous fetuses in the abdominal cavity of an elderly female dog with pyometra

This paper reports on a rare case of fetal papyraceous mummification after asymptomatic uterine rupture in an elderly female dog with pyometra. The patient had a history of mating six months before the examination but no apparent signs of gestation or parturition. Exploratory laparotomy was used to identify a rupture of the left uterine horn and the presence of cystic endometrial hyperplasia and pyometra. Two mummified papyraceous fetuses were observed in the abdominal cavity and had adhered to the spleen, pancreas, intestine and omentum. Ovariehysterectomy and corrective surgery were performed. The patient had remained healthy after uterine rupture until a new estrous cycle and the development of pyometra. Bitches that are 10 years old or more are predisposed to implantation failure, pregnancy or parturition problems and they should not be breed to avoid complications.

Transplacental Transfer of Iron in the Water Buffalo (Bubalus bubalis): Uteroferrin and Erythrophagocytosis

The objectives of this investigation were to understand transplacental transport of iron by secreted uteroferrin (UF) and haemophagous areas of water buffalo placenta and clarify the role(s) of blood extravasation at the placental-maternal interface. Placentomes and interplacentomal region of 51 placentae at various stages of gestation were fixed, processed for light and transmission electron microscopy, histochemistry and immunohistochemistry. Haemophagous areas were present in placentomes collected between 4 and 10 months of pregnancy. Perl's reaction for ferric iron was negative in placentomes, but positive in endometrial glands. Positive staining for UF indicated areas in which it was being taken up by phagocytosis and/or fluid phase pinocytosis in areolae of the interplacentomal mesenchyme, with little staining in endometrial stroma. Imunohistochemistry detected UF in trophectoderm of haemophagous regions of placentomes and in other parts of the foetal villous tree, but the strongest immunostaining was in the epithelial cells and lumen of uterine glands. Ultrastructural analyses indicated that erythrophagocytosis was occurring and that erythrocytes were present inside cells of the chorion that also contained endocytic vesicles and caveolae. Results of this study indicate that both the haemophagous areas of placentomes and the areolae at the interface between chorion and endometrial glands are important sites for iron transfer from mother to foetal-placental tissues in buffalo throughout pregnancy.

Morpho-physical Recording of Bovine Conceptus (Bos indicus) and Placenta from Days 20 to 70 of Pregnancy

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

Bone healing in critical-size defects treated with platelet-rich plasma: a histologic and histometric study in rat calvaria

Background and objective: The purpose of this study was to analyze histologically the influence of autologous platelet-rich plasma on bone healing in surgically created critical-size defects in rat calvaria.Material adn Methods: Thirty-two rats were divided into two groups: the control group (group C) and the platelet-rich plasma group. An 8-mm-diameter critical-size defect was created in the calvarium of each animal. In group C the defect was filled by a blood clot only. In the platelet-rich plasma group, 0.35 mL of platelet-rich plasma was placed in the defect and covered by 0.35 mL of platelet-poor plasma. Both groups were divided into subgroups (n = 8) and killed at either 4 or 12 wk postoperatively. Histometric (using image-analysis software) and histologic analyses were performed. The amount of new bone formed was calculated as a percentage of the total area of the original defect. Percentage data were transformed into arccosine for statistical analysis (analysis of variance, Tukey, p < 0.05).Results: No defect completely regenerated with bone. The platelet-rich plasma group had a statistically greater amount of bone formation than group C at both 4 wk (17.68% vs. 7.20%, respectively) and 12 wk (24.69% vs. 11.65%, respectively) postoperatively.Conclusion: Within the limits of this study, it can be concluded that platelet-rich plasma placed in the defects and covered by platelet-poor plasma significantly enhanced bone healing in critical-size defects in rat calvaria.

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.

Healing outcomes at implants installed in grafted sites: an experimental study in dogs

Aim: To evaluate the integration of implants installed using a surgical guide in augmented sites with autologous bone or deproteinized bovine bone mineral (DBBM) blocks, concomitantly with a collagen membrane.Material and methods: Mandibular molars were extracted bilaterally in six Labrador dogs, the buccal bony wall was removed, and a box-shaped defect was created. After 3 months, flaps were elevated, a bony graft was harvested from the ascending ramus, and secured to the lateral wall of the defect by means of screws. In the left mandibular side, a DBBM block was fixed into the defect. A resorbable membrane was applied at both sides, and the flaps were sutured. After 3 months, flaps were elevated, and a customized device was used as surgical guide to prepare the recipient sites in the interface between grafts and parent bone. One implant was installed in each side of the mandible. After 3 months, biopsies were harvested, and ground sections were prepared for histologic evaluation.Results: One autologous bone block graft was lost before implant installation. The width of the alveolar crest at the test sites (DBBM) was 5.4 +/- 1.2 mm before, 9.4 +/- 1.2 mm immediately after grafting, and 9.3 +/- 1 mm at implant installation. At the control sites (autologous bone), the corresponding values were: 5.2 +/- 1, 9 +/- 1.2, and 8.7 +/- 0.9 mm, respectively. All implants installed were available for histologic evaluation (n = 5). The autologous bone grafts, rich in vessels and cells, were integrated in the parent bone, and only little non-vital bone was found. The BIC% was 56.7 +/- 15.6% and 54.2 +/- 13.2% at the buccal and lingual aspects, respectively. At the test sites, the DBBM appeared to be embedded into connective tissue, and very little newly formed bone was encountered within the grafts. The BIC% was 5.8 +/- 12.3% and 51.3 +/- 14.2% at the buccal and lingual aspects, respectively.Conclusions: Autologous bone blocks used to augment the alveolar bony crest horizontally allowed the complete osseointegration of implants installed after 3 months of healing. However, similar blocks of DBBM did not promote osseointegration, although the installed implants were stable owing to the osseointegration in the sites of the parent bone.

Rat nephrotoxic nephritis. The relation between the type and intensity of induced histological lesions and the content of antiglomerular basement membrane antibodies of the inoculated serum

Six groups of 6 rats received equal doses (0.8 ml/100 g of body weight) of different rabbit anti rat kidney sera. The titer of anti GBM antibodies in the sera was evaluated by indirect immunofluorescent test in isolated GBM (IIT GBM). Rats of groups 1, 2, 3, 5, 6 received anti rat GBM sera with titers of 1/320, 1/240, 1/160, 1/60, 1/30 respectively. Group 4 received anti rat kidney serum with a titer of 1/80. The rats of group 1 died from 1 to 5 minutes after inoculation and their kidney were congested, with hialine trombi occluding arterioles and glomerular capillaries. The rats of group 2 and one of group 3 died from 2 to 15 days after inoculation and diffuse cortical necrosis was found. The remaining rats were sacrificed 2 months after inoculation. The kidneys were normal in control group; chronic membranoproliferative glomerulonephritis was observed in group 3 and 4, membranoproliferative glomerulonephritis in group 5 and minimal changes in group 6. By immunofluorescence rabbit gammaglobulin was seen in GBM of group 3, 4, 5 and 6. The IIT GBM performed in the eluates of the kidneys revealed the presence of heterologous antibody in groups 1, 2, 3, 4, 5 and 6 and autologous antibody in groups 3, 4 and 5. One concludes that the IIT GBM identifies and quantifies antibodies which have the property of damaging the kidney.

Treatment of rat nephrotoxic nephritis. Use of 5-fluorouracil or methotrexate-5-fluorouracil association

To evaluate the effect of 5-fluorouracil (F) and methotrexate-5-fluorouracil association (MTX-F) on nephrotoxic nephritis, seven groups of 10 rats were inoculated with anti-rat glomerular basement membrane serum (AGBMS); five groups were treated with different doses of F, beginning on the 2nd or the 6th day, one group with MTX-F beginning on the 2nd day and one group (control) with distilled water. Twenty-four hour proteinuria was determined weekly until the 71st day. The kidneys were examined histologically and by immunofluorescence. The group treated with F (1.3 mg/100 g body weight) developed a severe glomerulonephritis similar to the control group; (b) the groups treated with F (2.0 mg/100 g body weight) or with MTX-F showed progressively lower proteinuria, less severe histological changes and less intense fluorescence due to autologous antibodies. The best results were observed in the MTX-F group and in the F group treated from the 6th day. These groups presented at the 71st day proteinuria of 84 and 91 mg as compared to 312 mg in the control group, and minimal histological lesions as compared to glomerulosclerosis and tubular atrophy in the control group. We concluded that either F or MTX-F produced significant improvement of nephrotoxic nephritis due to inhibition of autologous antibody production.

Inside-out vs. standard vein graft to repair a sensory nerve in rats

Nerve regeneration in a sensory nerve was obtained by the application of different techniques: inside-out vein graft (IOVG group) and standard vein graft (SVG group). These techniques provide a good microenvironment for axon regeneration in motor nerves, but their efficiency for regeneration of sensory nerves is controversial. The saphenous nerve was sectioned and repaired by the inside-out and standard vein graft techniques in rats. After 4, 12, and 20 weeks the graft and the distal stump were observed under electron microscopy. In each studied period, the pattern, diameters, and thickness of the myelin sheaths of the regenerated axons were measured in the graft and distal stump. A comparative study about the regenerated nerve fibers by these two different techniques was performed. Regenerated nerve fibers were prominent in both vein grafts 4 weeks after the surgical procedures. On the other hand, in the distal stump, regenerated nerve fibers were observed only from 12 weeks. In both inside-out vein graft and standard vein graft statistical difference was not observed about the diameters and thickness of the myelinated fibers after 20 weeks. On the other hand, the inside-out group had greater regenerated axon number when compared to the standard group. There is a capillary invasion in both graft and distal stump, especially in the IOVG group. The regenerated axons follow these capillaries all the time like satellite microfascicles. After 20 weeks, the diameters of regenerated fibers repaired by the standard vein graft technique were closer to the normal fibers compared to the inside-out vein graft. On the other hand, the pattern of these regenerated axons was better in the IOVG group.