Expressão gênica de bcl2, receptor de estradiol e receptores de progesterona A e B em endométrio eutópico e ectópico de pacientes inférteis sem e com endometriose

A endometriose caracteriza-se pela presença de tecido endometriótico – glândulas e estroma – fora da cavidade uterina. A doença acomete superfícies peritoniais da pélvis, ovários e septo rectovaginal, sendo mais comum a endometriose pélvica. Embora os dados sobre a prevalência exata de endometriose sejam incertos, as evidências indicam uma prevalência em torno de 5 a 10% das mulheres em idade reprodutiva. Dor, dismenorréia, dispareunia e infertilidade são sintomas freqüentes da doença. Além de apresentar dependência aos hormônios sexuais, a endometriose está associada a um conjunto de desordens de diferentes etiologias, sendo considerada uma doença multifatorial de caráter genético e imune. Neste sentido, a carência de respostas imunológicas adequadas, aliada a um provável desequilíbrio entre os processos de proliferação e apoptose celular, pode ter um papel importante na instalação e manutenção das lesões endometrióticas. O objetivo deste trabalho foi caracterizar a expressão de genes relacionados aos processos de proliferação e diferenciação celular em implantes endometrióticos e endométrio eutópico de mulheres inférteis sem e com endometriose, através da análise semiquantitativa por RT-PCR. Foram estudadas 34 pacientes consultando por infertilidade e submetidas à laparoscopia diagnóstica. Quinze pacientes (31,7 ± 1,5 anos) apresentavam endometriose e 19 (32,9 ± 0,9 anos) foram consideradas controles inférteis sem endometriose. Os dois grupos apresentaram IMC e SHBG similares. As biópsias de endométrio foram realizadas, em fase folicular, nas pacientes do grupo controle (C) e nas pacientes com endometriose (endométrio eutópico (EUT) e ectópico (ECT)). A presença do mRNA para os receptores de estrogênio e progesterona detectada nos 3 grupos estudados, sustenta a existência de sensibilidade tecidual local aos hormônios sexuais. Nas condições experimentais do presente estudo não foi observada diferença estatística na expressão gênica de bcl2 entre os grupos. A expressão do gene do ER endometrial também foi similar entre os grupos controle, eutópico e ectópico. Contudo, o endométrio ectópico apresentou níveis de mRNA mais elevados para PR-A (0,84  0,02 UA) em relação aos grupos controle e eutópico (0,60  0,04 UA e 0,63  0,04 UA; p < 0,05). A expressão gênica do PR-B não foi estatisticamente diferente entre os grupos do presente trabalho. Porém, houve forte correlação negativa entre a expressão dos genes bcl2 e PR-B nas amostras de endométrio sem e com endometriose (r = - 0,795 e p = 0,018), sugerindo que um papel anti-proliferativo do PR-B possa ser operativo neste modelo de estudo.

Purificação e caracterização parcial de duas n-acetil-B-hexosaminidases do equinoderma marinho echinometra lucunter

Two b-N-acetylhexosaminidases (F11 e F15) were purified from Echinometra lucunter gonads extracts. The purified enzymes were obtained using ammonium sulfate fractionation, followed by gel filtration chromatographies (Sephacryl S-200, Sephadex G-75 and Sephacryl S-200). The F11 fraction was purified 192.47 -fold with a 28.5% yield, and F15 fraction 85.41 -fold with a 32.3% yield. The molecular weights of the fractions were 116 kDa for F11 and 42 kDa for F15 using SDS-PAGE. In Sephacryl S-200, F15 was 84 kDa, indicating that it is a dimeric protein. When p-nitrophenyl-β-D-glycosaminide was used as substrate, we determined an apparent Km of 0.257 mM and Vmax of 0.704 for F11 and for F15 the Km was 0.235 mM and Vmax of 0.9 mM of product liberated by hour. Both enzymes have optimum pH and temperature respectively at 5.0 and 45 °C. The enzymes showed inhibition by silver nitrate, while the glucuronic acid was a potent activator. The high inhibition of F15 by N-etylmaleimide indicates that sulphydril groups are involved in the catalysis of synthetic substrate

Caracterização estrutural e avaliação das atividades farmacológicas da fucana B extraída da alga Dictyota menstrualis

Seaweeds are a major source of biologically active compounds . In the extracellular matrix of these organisms are sulfated polysaccharides that functions as structural components preventing it against dehydration. The fraction 0.9 (FucB) rich in sulfated fucans obtained from brown seaweed Dictyota menstrualis was chemical characterized and evaluated for pharmacological activity by testing anticoagulant activity, stimulatory action on the synthesis of an antithrombotic heparan sulfate, antioxidant activity and its effects in cell proliferation. The main components were FucB carbohydrates (49.80 ± 0.10 %) and sulfate (42.30 ± 0.015 %), with phenolic compounds ( 3.86 ± 0.016 %) and low protein contamination ( 0.58 ± 0.001 % ) . FucB showed polydisperse profile and analysis of signals in the infrared at 1262, 1074 and 930 cm -1 and 840 assigned to S = O bonds sulfate esters , CO bond presence of 3,6- anhydrogalactose , β -D- galactose non- sulfated sulfate and the axial position of fucose C4 , respectively. FucB exhibited moderate anticoagulant activity , the polysaccharides prolonged time (aPTT ) 200 ug ( > 90s ) partial thromboplastin FucB no effect on prothrombin time (PT), which corresponds to the extrinsic pathway of coagulation was observed. This stimulation promoted fraction of about 3.6 times the synthesis of heparan sulfate (HS) by endothelial cells of the rabbit aorta ( RAEC ) in culture compared with cells not treated with FucB . This has also been shown to compete for the binding site with heparin. The rich fraction sulfated fucans exhibited strong antioxidant activity assays on total antioxidant (109.7 and 89.5 % compared with BHT and ascorbic acid standards ) , reducing power ( 71 % compared to ascorbic acid ) and ferric chelation ( 71 , comparing with 5 % ascorbic acid). The fraction of algae showed cytostatic activity on the RAEC cells revealed that the increase of the synthesis of heparan sulfate is not related to proliferation. FucB showed antiproliferative action on cell lines modified as Hela and Hep G2 by MTT assay . These results suggest that FucB Dictyota menstrualis have anticoagulant , antithrombotic , antioxidant potential as well as a possible antitumor action, promoting the stimulation of the synthesis of antithrombotic HS by endothelial cells and is useful in the prevention of thrombosis, also due to its inhibitory action on species reactive oxygen ( ROS ) in some in vitro systems , being involved in promoting a hypercoagulable state

Purification and preliminary crystallographic analysis of a new Lys49-PLA2 from B-jararacussu

BjVIII is a new myotoxic Lys49-PLA2 isolated from Bothrops jararacussu venom that exhibits atypical effects on human platelet aggregation. To better understand the mode of action of BjVIII, crystallographic studies were initiated. Two crystal forms were obtained, both containing two molecules in the asymmetric unit (ASU). Synchrotron radiation diffraction data were collected to 2.0 angstrom resolution and 1.9 angstrom resolution for crystals belonging to the space group P2(1)2(1)2(1) (a = 48.4 angstrom, b = 65.3 angstrom, c = 84.3 angstrom) and space group P3(1)21 (a = b = 55.7 angstrom, c = 127.9 angstrom), respectively. Refinement is currently in progress and the refined structures are expected to shed light on the unusual platelet aggregation activity observed for BjVIII.

Evolução de títulos sorológicos, nas provas de soroaglutinação em placa, antígeno acidificado tamponado e fixação de complemento, em bezerras Nelore vacinadas aos 18 meses de idade com Brucella abortus amostra B 19

Com o objetivo de estudar a persistência da resposta sorológica, através das provas de soroaglutinação em placa, rosa Bengala e fixação de complemento, 108 bezerras, com idade ao redor de 18 meses, foram vacinadas com uma dose padrão da vacina preparada com Brucella abortus amostra B 19. Foram obtidas amostras de soro sangüíneo antes da vacinação e após 45 dias, 6, 12 e 18 meses. Antes da vacinação, todas as bezerras apresentaram resultados negativos nos três testes. Após 45 dias, todas apresentaram título a partir de 1/100 na prova de soroaglutinação em placa e todas apresentaram resultado positivo no teste rosa Bengala, ao passo que dois animais apresentaram título 1/2 e os demais apresentaram título a partir de 1/4 na reação de fixação de complemento. Após um ano de vacinação, a grande maioria das bezerras já não apresentava título sorológico significativo. Considerando o risco a que estão sujeitos animais que vivem em áreas endêmicas e em propriedades onde a doença ocorre, e considerando a acentuada redução de título sorológico observada na grande maioria dos animais, pode-se concluir que, no caso bezerras de raças zebuínas em áreas endêmicas e que não tenham sido vacinadas na idade regulamentar, é mais vantajoso vaciná-las com a amostra B 19 aos 18 meses de idade do que deixá-las expostas a um elevado risco de infecção por Brucella.

Structural and functional evidence that a B chromosome in the characid fish Astyanax scabripinnis is an isochromosome

Astyanax scabripinnis possesses a widespread polymorphism for metacentric B chromosomes as large as the largest chromosome pair in the A complement. on the basis of C-banding pattern, it was hypothesized that these B chromosomes are isochromosomes that have arisen by means of centromere misdivision and chromatid nondisjunction. In the present paper we test this hypothesis by analysing (i) the localization of a repetitive DNA sequence on both B chromosome arms, and (ii) synaptonemal complex formation, in order to test the functional homology of both arms. Genomic DNA digested with KpnI and analysed by gel electrophoresis showed fragments in a ladder-like pattern typical of tandemly repetitive DNA. These fragments were cloned and their tandem organization in the genome was confirmed. A 51-bp long consensus sequence, which was AT-rich (59%) and contained a variable region and two imperfect reverse sequences, was obtained. Fluorescence in situ hybridization (FISH) localized this repetitive DNA into noncentromeric constitutive heterochromatin which encompasses the terminal region of some acrocentric chromosomes, the NOR region, and interstitial polymorphic heterochromatin in chromosome 24. Most remarkably, tandem repeats were almost symmetrically placed in the two arms of the B chromosome, with the exception of two additional small clusters proximally located on the slightly longer arm. Synaptonemal complex (SC) analysis showed 26 completely paired SCs in males with 1B. The ring configuration of the B univalent persisting until metaphase I suggests that the two arms formed chiasmata. All these data provided strong support for the hypothesis that the B chromosome is an isochromosome.

Expressão imuno-histoquimica de colagenase-1 e gelatinases A e B em mixomas odontogênicos e papilas de germes dentários

The odontogenic myxoma shares cellular and structural aspects with dental papilla, which has been implicated as probable origin of this neoplasm. The aim of the present study was to perform a comparative immunohistochemical analysis for the expression of collagenase-1 (MMP-1) and gelatinases A (MMP-2) and B (MMP-9) in odontogenic myxomas and dental papilla of teeth germs. Twelve cases of odontogenic myxomas and eight specimens of teeth germs were selected. It was taken into consideration the presence or absence of immunoreactivity, the pattern of immunohistochemical distribution of proteases within extracellular matrix, as well as, the number of cells revealing immunostaining for matrix metalloproteinases (MMPs). It was verified a significant difference (p<0,05) in relation to MMP-2 immunoexpression, which was observed only within extracellular matrix of myxomas. Nevertheless, MMP-1 labeling was revealed by most of the cases of odontogenic myxoma, at levels close to those observed in dental papilla. In relation to the pattern of distribution, a significant difference was obtained between specimens (p<0,05), with neoplasms predominantly exhibiting a focal pattern for MMP-1. The quantitative analysis of neoplastic cells labeled for MMPs denoted a significant difference (p<0,05), demonstrating a higher proportion of MMP-1 in comparison to MMPs-2 and -9. It can be concluded that immunohistochemical expression of MMP-1 at levels comparable to those observed in dental papilla and quantitatively superior in relation to MMPs-2 and -9, suggest an implication of this protease on extracellular matrix degradation of odontogenic myxomas. Moreover, the possibility of interactions with receptors involved in cellular adhesion, particularly with integrins, suggests a plausible function on local invasiveness of such neoplasms. Additionally, the presence of a descent immunoexpression gradient for these MMPs on odontogenic myxomas, associated to substrate specificity inherent in each enzyme, suggest the existence of a coordinated mechanism between interstitial collagenase and gelatinases A and B in order to allow an efficient degradation of extracellular matrix and local invasion by neoplastic cells

Tobacco smoke-induced left ventricular remodelling is not associated with metalloproteinase-2 or -9 activation

Aim: To investigate the role of MMP-2 and MMP-9 in cardiac remodelling induced by tobacco smoke exposure in rats.Methods: Rats were allocated into two groups: C (n = 9): control animals; ETS (n = 9): exposed to tobacco smoke. After 4months, the animals underwent echocardiography, morphometric study and determination of MMP-2 and MMP-9 activity.Results: ETS rats had larger diastolic (C= 15.6 +/- 1.2 mm/kg, ETS = 18.0 +/- 0.9 mm/kg; p < 0.001) and systolic (C= 7.3 +/- 1.2 mm/kg, ETS = 9.2 0.9 mm/kg; p = 0.001) ventricular diameters adjusted for body weight. Fractional shortening (C= 53 +/- 4.8%, ETS = 48 +/- 3.3%; p = 0.031) and ejection fraction (C= 0. 89 +/- 0.03 5 ETS = 0. 86 +/- 0.02; p = 0.03 0) were smaller in the ETS group. Myocyte cross-sectional area (C= 245 8 mu m(2), ETS=253 8 mu m(2); p = 0.028) was higher in ETS rats. There were no differences in MNtP-2 (C=50 +/- 14%; ETS 43 +/- 11%, p 0.22 +/- 8) or MMP-9 (C=0.36 +/- 0.3%; ETS=0.62 +/- 0.3%, p=0.630) activity between the groups.Conclusion: MMP-2 and MMP-9 did not participate in the remodelling process induced by tobacco smoke exposure. (c) 2007 European Society of Cardiology. Published by Elsevier B.V. All rights reserved.

Gravitomagnetismo e o teste da sonda gravidade B

The so-called gravitomagnetic field arised as an old conjecture that currents of matter (no charges) would produce gravitational effects similar to those produced by electric currents in electromagnetism. Hans Thirring in 1918, using the weak field approximation to the Einsteins field equations, deduced that a slowly rotating massive shell drags the inertial frames in the direction of its rotation. In the same year, Joseph Lense applied to astronomy the calculations of Thirring. Later, that effect came to be known as the Lense- Thirring effect. Along with the de Sitter effect, those phenomena were recently tested by a gyroscope in orbit around the Earth, as proposed by George E. Pugh in 1959 and Leonard I. Schiff in 1960. In this dissertation, we study the gravitational effects associated with the rotation of massive bodies in the light of the Einsteins General Theory of Relativity. With that finality, we develop the weak field approximation to General Relativity and obtain the various associated gravitational effects: gravitomagnetic time-delay, de Sitter effect (geodesic precession) and the Lense-Thirring effect (drag of inertial frames). We discus the measures of the Lense-Thirring effect done by LAGEOS Satellite (Laser Geodynamics Satellite) and the Gravity Probe B - GPB - mission. The GPB satellite was launched into orbit around the Earth at an altitude of 642 km by NASA in 2004. Results presented in May 2011 clearly show the existence of the Lense-Thirring effect- a drag of inertial frames of 37:2 7:2 mas/year (mas = milliarcsec)- and de Sitter effect - a geodesic precession of 6; 601:8 18:3 mas/year- measured with an accuracy of 19 % and of 0.28 % respectively (1 mas = 4:84810��9 radian). These results are in a good agreement with the General Relativity predictions of 41 mas/year for the Lense-Thirring effect and 6,606.1 mas/year for the de Sitter effect.

Group B streptococci colonization in pregnant women: risk factors and evaluation of the vaginal flora

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

Expressão de MMP-2 e MMP-9 no endométrio de éguas saudáveis e portadoras de endometrite crônica

Avaliaram-se, por método imunoistoquímico, a expressão e distribuição das metaloproteinases (MMP) 2 e 9 em amostras de endométrio hígido e de éguas portadoras de endometrite crônica. Foram utilizadas 60 biópsias endometriais. A MMP-2 foi observada na parede vascular, nas células estromais e no epitélio glandular, e a imunorreatividade mais intensa foi obtida nas células do epitélio glandular nas endometrites da categoria III e na parede vascular nos endométrios da categoria I. A marcação imunoistoquímica para MMP-9 mostrou-se difusa pelo endométrio e foi observada no epitélio luminal e glandular, na região da parede vascular, nas células estromais, endoteliais e do infiltrado inflamatório. Houve diminuição da marcação imunoistoquímica na região da parede vascular conforme aumentou o grau das lesões endometriais concomitante à diminuição da intensidade da reação. Não houve relação na expressão imunoistoquímica das metaloproteinases estudadas com o tipo de endometrite

PDGF-B gene therapy accelerates bone engineering and oral implant osseointegration

Platelet-derived growth factor-BB (PDGF-BB) stimulates repair of healing-impaired chronic wounds such as diabetic ulcers and periodontal lesions. However, limitations in predictability of tissue regeneration occur due, in part, to transient growth factor bioavailability in vivo. Here, we report that gene delivery of PDGF-B stimulates repair of oral implant extraction socket defects. Alveolar ridge defects were created in rats and were treated at the time of titanium implant installation with a collagen matrix containing an adenoviral (Ad) vector encoding PDGF-B (5.5 x 10(8) or 5.5 x 10(9) pfu ml (1)), Ad encoding luciferase (Ad-Luc; 5.5 x 10(9) pfu ml (1); control) or recombinant human PDGF-BB protein (rhPDGF-BB, 0.3 mg ml (1)). Bone repair and osseointegration were measured through backscattered scanning electron microscopy, histomorphometry, microcomputed tomography and biomechanical assessments. Furthermore, a panel of local and systemic safety assessments was performed. Results indicated that bone repair was accelerated by Ad-PDGF-B and rhPDGF-BB delivery compared with Ad-Luc, with the high dose of Ad-PDGF-B more effective than the low dose. No significant dissemination of the vector construct or alteration of systemic parameters was noted. In summary, gene delivery of Ad-PDGF-B shows regenerative and safety capabilities for bone tissue engineering and osseointegration in alveolar bone defects comparable with rhPDGF-BB protein delivery in vivo. Gene Therapy (2010) 17, 95-104; doi: 10.1038/gt.2009.117; published online 10 September 2009

Abnormal cell-cycle expression of the proteins p27, mdm2 and cathepsin B in oral squamous-cell carcinoma infected with human papillomavirus

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Association between PAX-9 promoter polymorphisms and hypodontia in humans

Hypodontia, the congenital absence of one or a few teeth, is one of the most common alterations of the human dentition. The most common permanent missing teeth are the third molars, second premolars, and maxitlary lateral incisors. Hypodontia does not represent a serious public health problem, but it may cause masticatory and speech dysfunctions, and esthetic problems. PAX9 is believed to play an important rote in tooth development. It is expressed at initiation, bud, cap, and bell stages of odontogenesis. Mutations in PAX9 coding sequences have been implicated in autosomal dominant oligodontia affecting predominantly permanent molars and second premolars. Here, we report two polymorphisms in the promoter region of PAX9 gene that are associated with hypodontia. DNA was extracted from buccal epithelial. cells of 106 healthy Control individuals and of 102 unrelated individuals with hypodontia. PCR-RFLP was employed in the investigation of G-1031A and T-912C polymorphisms. Significant differences were obtained comparing Control and Test groups. Alleles G and T were found at a significant higher frequency in individuals with hypodontia, whereas alleles A and C were more frequent in Control subjects, p = 0.0094 and 0.0086, respectively. The GT haplotype was significantly more prevalent in the hypodontia group, white the AC haplotype was more frequent in the Control group. These results indicate that polymorphisms in the promoter region of PAX9 gene may have an influence on the transcriptional activity of this gene and are associated with hypodontia in humans. (c) 2005 Elsevier Ltd. AR rights reserved.