Evaluation of left atrial function in asymptomatic dogs with chronic myxomatous mitral valve disease: two-dimensional feature-tracking echocardiography and Simpsons monoplanar methods / Avaliação da função atrial esquerda em cães assintomáticos com degeneração mixomatosa crônica de valva mitral: métodos ecocardiográficos bidimensional feature tracking e monoplanar de Simpson

The present study evaluated the volume and function of the left atrium by two-dimensional echocardiographic feature-tracking imaging (2D-FTI) and Simpson's monoplanar modeling in dogs with asymptomatic degenerative mitral valve disease (DMVD). The study consisted of 80 dogs that were divided into the following three groups: Group 1, 21 dogs (A); Group 2, 30 dogs (B1) and Group 3, 29 dogs (B2). The variable strain (contraction phase) was significantly lower in Group 3 than in Group 1 (12.92±4.54 x 16.69±5.74, p=0.014), and significant differences in the contraction strain index (CSI) were observed between all of the groups that were evaluated (1 = 46.82±8.10, 2 = 39.88±8.03, 3 = 35.25±5.64, p<0.0001). The atrial diastolic volume index (AdVi) that was measured by 2D-FTI was significantly higher in Group 3 than in Group 1 (1.31±0.95 x 0.96±0.31, p=0.038), and the atrial cardiac index (ACI) was also higher in Group 3 than in Group 1 (102.38±80.18 x 78.19±33.38, p=0.030). Atrial function was assessed by Simpson's monoplanar method, which demonstrated an increase in the left atrial systolic volume, while the contractile function decreased with an increasing disease severity (Group 1 0.21±0.06; Group 2 0.25±0.06; Group 3 0.32±0.08, p<0.0001). The intraobserver and interobserver assessments showed low to moderate variability; most of the values for the coefficient of variation for the variables that were analysed with each method were below 25%. Thus, DMVD was determined to cause an alteration in atrial function, especially in the contraction phase, and even in asymptomatic animals, and the methods of 2D-FTI echocardiography and Simpson's monoplanar evaluation are sensitive and early methods for the detection of left atrial dysfunction.(AU)

O presente estudo avaliou o volume e a função atrial esquerda obtidos por meio da ecocardiografia bidimensional feature tracking (2D-FTI) e pelo método monoplanar de Simpson em cães saudáveis e cães com DMVD assintomáticos. Foram avaliados 80 cães distribuídos em três grupos: Grupo 1, 21 cães (classe A); Grupo 2, 30 cães (classe B1) e Grupo 3, 29 cães (classe B2). A variável strain (fase de contração) foi significativamente menor no Grupo 3 que no Grupo 1 (12,92±4,54 x 16,69±5,74, p=0,014) e para a variável índice de strain de contração (CSI), houve diferença estatística entre todos os grupos avaliados (1 = 46,82±8,10; 2 = 39,88±8,03; 3 = 35,25±5,64, p<0,0001). O índice de volume diastólico atrial (iVdA) mensurado por meio do 2D-FTI foi significativamente maior no Grupo 3 que no Grupo 1 (1,31±0,95 x 0,96±0,31, p=0,038), assim como para o índice cardíaco atrial (iCA) também foi maior no Grupo 3 (102,38±80,18 x 78,19±33,38, p=0,030). A função atrial avaliada pelo método monoplanar de Simpson demonstrou um aumento do volume atrial esquerdo e do volume sistólico do átrio esquerdo, enquanto que a função contrátil diminuiu com o aumento da gravidade da doença (Grupo 1 0,21±0,06; Grupo 2 0,25±0,06; Grupo 3 0,32±0,08; p<0,0001). A avaliação intraobservador e interobservador, demonstrou variabilidade baixa a moderada, uma vez que a maioria dos valores de coeficiente de variação se concentraram abaixo de 25% para as variáveis analisadas em ambos os métodos. Dessa forma, conclui-se que a DMVD causa alteração na função atrial, principalmente na fase de contração, mesmo em animais assintomáticos e que a ecocardiografia 2D-FTI e o método monoplanar de Simpson são métodos sensíveis e precoces na detecção da disfunção atrial esquerda.(AU)

Expression of Hbs, Kirre, and Rst during Drosophila ovarian development.

The Irre cell-recognition module (IRM) is a group of evolutionarily conserved and structurally related transmembrane glycoproteins of the immunoglobulin superfamily. In Drosophila melanogaster, it comprises the products of the genes roughest (rst; also known as irreC-rst), kin-of-irre (kirre; also known as duf), sticks-and-stones (sns), and hibris (hbs). In this model organism, the behavior of this group of proteins as a partly redundant functional unit mediating selective cell recognition was demonstrated in a variety of developmental contexts, but their possible involvement in ovarian development and oogenesis has not been investigated, notwithstanding the fact that some rst mutant alleles are also female sterile. Here, we show that IRM genes are dynamically and, to some extent, coordinately transcribed in both pupal and adult ovaries. Additionally, the spatial distribution of Hbs, Kirre, and Rst proteins indicates that they perform cooperative, although largely nonredundant, functions. Finally, phenotypical characterization of three different female sterile rst alleles uncovered two temporally separated and functionally distinct requirements for this locus in ovarian development: one in pupa, essential for the organization of peritoneal and epithelial sheaths that maintain the structural integrity of the adult organ and another, in mature ovarioles, needed for the progression of oogenesis beyond stage 10.

Characterization of a novel Drosophila melanogaster cis-regulatory module that drives gene expression to the larval tracheal system and adult thoracic musculature.

In a previous bioinformatics analysis we identified 10 conserved Drosophila melanogaster sequences that reside upstream from protein coding genes (CGs). Here we characterize one of these genomic regions, which constitutes a Drosophila melanogaster cis-regulatory module (CRM) that we denominate TT-CRM. The TT-CRM is 646 bp long and is located in one of the introns of CG32239 and resides about 3,500 bp upstream of CG13711 and about 620 bp upstream of CG12493. Analysis of 646 bp-lacZ lines revealed that TT-CRM drives gene expression not only to the larval, prepupal, and pupal tracheal system but also to the adult dorsal longitudinal muscles. The patterns of mRNA expression of the transgene and of the CGs that lie in the vicinity of TT-CRM were investigated both in dissected trachea and in adult thoraces. Through RT-qPCR we observed that in the tracheal system the pattern of expression of 646 bp-lacZ is similar to the pattern of expression of CG32239 and CG13711, whereas in the thoracic muscles 646 bp-lacZ expression accompanies the expression of CG12493. Together, these results suggest new functions for two previously characterized D. melanogaster genes and also contribute to the initial characterization of a novel CRM that drives a dynamic pattern of expression throughout development.

Transcriptional cross-regulation of Irre Cell Recognition Module (IRM) members in the Drosophila pupal retina.

Cell adhesion molecules play a central role in morphogenesis, as they mediate the complex range of interactions between different cell types that result in their arrangement in multicellular organs and tissues. How their coordinated dynamic expression in space and time - an essential requirement for their function - is regulated at the genomic and transcriptional levels constitutes an important, albeit still little understood question. The Irre Cell Recognition Module (IRM) is a highly conserved phylogenetically group of structurally related single pass transmembrane glycoproteins belonging to the immunoglobulin superfamily that in Drosophila melanogaster are encoded by the genes roughest (rst), kin-of-irre (kirre), sticks-and-stones (sns) and hibris (hbs). Their cooperative and often partly redundant action are crucial to major developmental processes such axonal pathfinding, myoblast fusion and patterning of the pupal retina. In this latter system rst and kirre display a tightly regulated complementary transcriptional pattern so that lowering rst mRNA levels leads to a concomitant increase in kirre mRNA concentration. Here we investigated whether other IRM components are similarly co-regulated and the extent changes in their mRNA levels affect each other as well as their collective function in retinal patterning. Our results demonstrate that silencing any of the four IRM genes in 24% APF retinae changes the levels all other group members although only kirre and hbs mRNA levels are increased. Furthermore, expression, in a rst null background, of truncated versions of rst cDNA in which the portion encoding the intracellular domain has been partially or completely removed not only can still induce changes in mRNA levels of other IRM members but also result in Kirre mislocalization. Taken together, our data point to the presence of a highly precise and fine-tuned control mechanism coordinating IRM expression that may be crucial to the functional redundancy shown by its components during the patterning of the pupal retina.

Identification of Long Noncoding RNAs Deregulated in Papillary Thyroid Cancer and Correlated with BRAFV600E Mutation by Bioinformatics Integrative Analysis.

Papillary Thyroid Cancer (PTC) is an endocrine malignancy in which BRAFV600E oncogenic mutation induces the most aggressive phenotype. In this way, considering that lncRNAs are arising as key players in oncogenesis, it is of high interest the identification of BRAFV600E-associated long noncoding RNAs, which can provide possible candidates for secondary mechanisms of BRAF-induced malignancy in PTC. In this study, we identified differentially expressed lncRNAs correlated with BRAFV600E in PTC and, also, extended the cohort of paired normal and PTC samples to more accurately identify differentially expressed lncRNAs between these conditions. Indirectly validated targets of the differentially expressed lncRNAs in PTC compared to matched normal samples demonstrated an involvement in surface receptors responsible for signal transduction and cell adhesion, as well as, regulation of cell death, proliferation and apoptosis. Targets of BRAFV600E-correlated lncRNAs are mainly involved in calcium signaling pathway, ECM-receptor interaction and MAPK pathway. In summary, our study provides candidate lncRNAs that can be either used for future studies related to diagnosis/prognosis or as targets for PTC management.

Protocol for collection and separation of bone marrow mononuclear cells in chlorocebus aethiops / Protocolo para colheita e isolamento de células mononucleares de medula óssea em chlorocebus aethiops

Chlorocebus aethiops is a species of non-human primate frequently used in biomedical research. Some research involves this species as an experimental model for various diseases and possible treatment with stem cells. The bone marrow is one of the main sources of these cells and provides easy access. The aim of this study was to standardize the protocol of collection and separation of bone marrow in C. aethiops. Ten animals were submitted to puncture of bone marrow with access to the iliac crest and cell separation by density gradient. The bone marrow of C. aethiops had an average of 97% viability. From the results achieved, we can conclude that C. aethiops is an excellent model to obtain and isolate mononuclear cells from bone marrow, fostering several studies in the field of cell therapy.

Chlorocebus aethiops é uma espécie de primata não humano frequentemente utilizados em pesquisa biomédica. Algumas pesquisas envolve esta espécie como modelo experimental para várias doenças e possível tratamento com células-tronco. A medula óssea é uma das principais fontes destas células e proporciona fácil acesso. O objetivo deste estudo foi o de padronizar o protocolo de coleta e separação de medula óssea em C. aethiops. Dez animais foram submetidos a punção de medula óssea com acesso à crista ilíaca e separação de células por gradiente de densidade. A medula óssea de C. aethiops tinha uma média de 97% de viabilidade. A partir dos resultados obtidos, podemos concluir que C. aethiops é um excelente modelo para obter e isolar células mononucleares da medula óssea, promovendo vários estudos no campo da terapia celular.

Amplification and expression of a salivary gland DNA puff gene in the prothoracic gland of Bradysia hygida (Diptera: Sciaridae).

The DNA puff BhC4-1 gene, located in DNA puff C4 of Bradysiahygida, is amplified and expressed in the salivary gland at the end of the fourth larval instar as a late response to the increase in 20-hydroxyecdysone titer that triggers metamorphosis. Functional studies revealed that the mechanisms that regulate BhC4-1 expression in the salivary gland are conserved in transgenic Drosophila. These studies also led to the identification of a cis-regulatory module that drives developmentally regulated expression of BhC4-1-lacZ in the prothoracic gland cells of the ring gland, a compound organ which in Drosophila results from the fusion of the prothoracic glands, the corpus allatum and the corpus cardiacum. Here we have investigated the occurrence of BhC4-1 expression in B. hygida prothoracic glands. We report the identification of the B. hygida prothoracic gland and demonstrate that it releases ecdysone. Using RT-qPCR, western blots and immunolocalization experiments, we demonstrate that the BhC4-1 mRNA and the BhC4-1 protein are both expressed in the B. hygida prothoracic glands at the same time that DNA puff C4 is formed in the salivary gland. We also show that BhC4-1 is concomitantly amplified 4.8-fold in the prothoracic gland and 23-fold in the salivary gland. Our results reveal the occurrence of stage specific expression of a DNA puff gene in the prothoracic glands of B. hygida, and extend previous studies that have shown that DNA puff genes expression is not restricted to the salivary gland. In addition, the description of stage specific gene amplification in the prothoracic glands of B. hygida constitutes the first demonstration that gene amplification in Diptera might occur concomitantly in two different tissues in the same developmental stage.

Morfologia e morfometria da traqueia da preguiça (bradypus variegatus): conhecimentos para procedimentos de emergência / Morphological and morphometric trachea sloth (bradypus variegatus): knowledge for emergency procedures

Por ser uma espécie pouco estudada, principalmente do ponto de vista morfológico, objetivou-se conhecer a anatomia da traqueia da preguiça (Bradypus variegatus) a fim de fornecer informações para facilitar a eleição de tubo endotraqueal adequado, máscara laríngea ou cânula de traqueostomia para anestesia e procedimentos de emergência, uma vez que a mesma revelou-se possuidora de uma morfologia especial. Foram investigados 11animais jovens de idades diferentes, sendo quatro machos e sete fêmeas, provenientes do Museu Emilio Goeldi e doados a UFRA. Os exemplares foram perfundidos via intramuscular com solução aquosa de formol a 10% para fins de conservação e posteriormente foram dissecados em nível cervico-torácico, por meio de mesoscopia, expondo-se desde a laringe até os brônquios principais direito e esquerdo ao nível do hilo pulmonar. As traqueias foram divididas em cinco regiões (região cervical, região da primeira flexura, região da segunda flexura, região da terceira flexura e região da carina) as quais foram mensuradas altura e largura, assim como o comprimento traqueal total e parte do material foi submetido à técnicas de rotina histológica. Macroscopicamente, destacou-se a presença de sinuosidades acentuadas em porção médio caudal, contemplando a carina. O comprimento médio traqueal foi de 14,6cm. Microscopicamente, a traqueia era constituída por placas separadas de cartilagem hialina constituindo cada anel, sendo revestido por epitélio estratificado ciliado. Apesar da traqueia da preguiça comum apresentar revestimento padrão encontrado na traqueia de outros animais, na literatura não há registros de outras espécies que tenham morfologia macroscópica nas condições descritas, o que nos leva a sugerir, quando necessário acesso para ventilação de emergência, a prática da IOT e não a de traqueostomia.

As the sloth (Bradypus variegatus) is a little studied species, especially from a morphological point of view, this research aimed to define the anatomy of its trachea. The information would facilitate the selection of a proper endotracheal tube, laryngeal mask or tracheostomy tube for anesthesia and emergency procedures, since it appeared to have a special morphology. Eleven young animals of different ages were investigated, four males and seven females, obtained from the Emilio Goeldi Museum and donated to UFRA. The specimens were infused intramuscularly with 10% aqueous formaldehyde for preservation and were later dissected at the cervico-thoracic level, by mesoscopia, exposing the area from the larynx to the right and left primary bronchi at the hilum. The tracheae were divided into five regions (cervical, first flexure, second flexure, third flexure, and carina) for which length and width were measured, as well as the total tracheal length. Sharp windings were seen in the middle caudal portion, including the carina. The average tracheal length was 14.6 cm. Microscopically, the trachea was made up of separate plates of hyaline cartilage forming each ring, lined with ciliated epithelium. Despite the trachea of the common sloth displaying the same lining pattern found in other animals, there are no reports in the literature of other species having a macroscopic morphology as described here, which leads us to suggest, where appropriate access to emergency ventilation, the practice of IOT and not tracheostomy.

Prenatal development of the agouti (Dasyprocta prymnolopha Wagler, 1831): External features and growth curves.

The gestation period in agoutis can range from 104 to 120 days. Knowledge regarding the morphological characteristics of embryos and fetuses is important as a base for studies in reproduction biotechnology, such as in vitro fertilization, embryo transfer and helps in determining congenital anomalies during the development phase. Thus, given the importance and lack of information about agouti embryology, the objective of this study was to characterize the external morphology and define the biometry of embryos and fetuses, at different days of development. Nine females were submitted to daily colpocytology to identify the estrus, confirm mating and identify day zero of the gestation. When the mating was confirmed they were weighed, underwent abdominal ultrasonography and surgery was conducted on the females at the gestational ages of 25, 30, 35, 40, 45, 50, 75 and 100 days. Sixteen embryos/fetuses were weighed and measured. Agouti embryos at 25 days after mating are "C" shaped, with primitive structures, 0.4±0.01cm crown-rump and weighed 0.06±0.01g; at 30 days after mating the crown-rump was 0.95±0.07cm and weighed 0.28±0.00g; at 35 days after mating the crown-rump was 155±0.07cm and weighed 0.38±0.01g; at 40 days after mating the crown-rump was 2.25±0.21cm and weighed 1.25±0.07g; at 45 days after mating the crown-rump was 3.45±0.35cm and weighed 2.75±0.64g; at 50 days after mating the crown-rump was 5.0±0.3cm and weighed 7.01±2.6g; at 75 days after mating, the skin was dark, the crown-rump was 10.0±0.14cm and weighed 55.2±0.07g. At 100 days after mating, the crown-rump was 13.8±0.49cm and fetuses weighed 136.7±9.40g. Based on the morphological data assessed the embryo and fetus age could be assessed and the size and average weight of agouti embryos was established.

Modelos experimentais de doenças pulmonares fibrosantes / Experimental models of fibrotic lung diseases

As doenças pulmonares intersticiais fibrosantes são enfermidades graves e de etiologia pouco conhecida. A necessidade de encontrar modelos experimentais que possam reproduzir essas doenças pulmonares desde os estágios iniciais até as fases avançadas levou pesquisadores a testar inúmeros modelos biológicos para esse fim. Diferentes modelos experimentais são usados para o estudo de doenças pulmonares, como murinos, suínos, em coelhos e em primatas não humanos. O objetivo da presente revisão foi apresentar os modelos experimentais mais utilizados. Atualmente, os modelos experimentais mais utilizados para o estudo das afecções pulmonares agudas e crônicas são os pequenos roedores e os miniporcos. Elaborar modelos experimentais significa enfrentar dificuldades relacionadas às diferenças fisiológicas, anatômicas e mecânicas dos sistemas orgânicos entre humanos e a espécie eleita. A relação custo-benefício baseia-se na facilidade para se reproduzir a doença pretendida, os custos do biotério e a semelhança entre os achados clínico-histopatológicos do modelo experimental com humanos. Sendo assim, não existe modelo experimental perfeito, mas modelos adequados para proceder à investigação científica almejada.

Anatomy and morphometric aspects of the trachea of Saimiri sciureus Linnaeus, 1758: knowledge for emergency procedures.

The aim of this work is to study the morphological characteristics of the trachea of Saimiri sciureus through quantification and measurement of the cartilaginous rings, providing information to facilitate the election of more appropriate endotracheal tube, laryngeal mask or tracheostomy tube for anesthetic and emergency procedures, as it is a species of Neotropical primates most commonly used as biological models, and little is known about their morphology. Nine animals were investigated, being 4 adults and 5 young acquired from the Centro Nacional de Primatas (National Primate Center - CENP) - Ananindeua - PA, which died from natural causes and then fixed in aqueous buffered formalin 10%. Saimiri sciureus trachea comprises an average of 32.8 incomplete rings and an average length of 3.74 cm in young animals, while in adults it demonstrated an average of 30.25 rings and average length of 3.67 cm. The shape of the light and its proportion varied along the trachea. Endotracheal tube with a diameter the 2.0 - 2.5 mm, laryngeal mask number 1.0 or tracheostomy tube neonatal Shiley number 3.0, can be placed in animals weighing 600 g - 1.2 Kg. Given the great importance of the species studied, which is widely used as a biological model, the detailing on the morphology and morphometry of tracheal animal studies provides new approaches needed in respiratory emergency, as well as, facilitates the development of future anesthetic protocols.

Kidney injury and cell therapy: Preclinical study.

The aim of this study is to show histological and immunofluorescence analysis of renal parenchyma of agoutis affected by gentamicin-induced renal disease after the infusion of bone marrow mononuclear cells (BMMC) stained with Hoechst®. Nine agouti's males were divided into three groups: Test group (TG): renal disease by gentamicin induced (n = 3), cell therapy group (CTG): renal disease by gentamicin induced and BMMC infusion (n = 3), and control group (CG): nonrenal disease and BMMC infusion (n = 3). TG and CTG were submitted to the protocol of renal disease induction using weekly application of gentamicin sulfate for 4 months. CG and CTG received a 1 × 108 BMMC stained with Hoechst and were euthanized for kidney examination 21 days after BMMC injection and samples were collected for histology and immunofluorescence analysis. Histological analysis demonstrated typical interstitial lesions in kidney similarly to human disease, as tubular necrosis, glomerular destruction, atrophy tubular, fibrotic areas, and collagen deposition. We conclude that histological analysis suggest a positive application of agouti's as a model for a gentamicin inducing of kidney disease, beyond the immunofluorescence analysis suggest a significant migration of BMMC to sites of renal injury in CTG.

Bone marrow cells of swine: collection and separation.

Bone marrow is a source of stem cells for greater and easier access, which is widely studied as a provider of hematopoietic and mesenchymal cells for various purposes, mainly therapeutic by the advances in research involving cell therapy. The swine is an animal species commonly used in the pursuit of development of experimental models. Thus, this study aimed to standardize protocol for collection and separation of bone marrow in swines, since this species is widely used as experimental models for various diseases. Twelve animals were used, which underwent bone marrow puncture with access from the iliac crest and cell separation by density gradient followed by a viability test with an average of 98% of viable cells. Given our results, we can ensure the swine as an excellent model for obtaining and isolation of mononuclear cells from bone marrow, stimulating several studies addressing the field of cell therapy.

Cell therapy for fibrotic interstitial pulmonary disease: experimental study.

Parte superior do formulário Digite um texto ou endereço de um site ou traduza um documento. The aim of this study is to evaluate the histological changes in lung parenchyma of pigs affected by interstitial lung disease induced after the infusion of bone marrow mononuclear cells (BMMCs). Ten female swines were submitted to pulmonary fibrosis induced by a single dose of intratracheal bleomicine sulfate. Animals were arranged into two groups: Group 1: induced-disease control and Group 2: cell therapy using BMMCs. Both groups were clinically evaluated for 180 days. High-resolution computed tomography (HRCT) was performed at 90 and 180 days. BMMC sampling was performed in cell therapy group at 90 days. Euthanasia was performed, and samples were collected for histology and immunohistochemistry. The 90-days HRCT demonstrated typical interstitial lesions in pulmonary parenchyma similarly to human disease. The 180-days HRCT in Group 1 demonstrated advanced stages of the disease when compared with Group 2. Immunohistochemistry analysis suggests the presence of pre-existent vessels and neoformed vessels as well as predominant young cells in the injured parenchyma of Group 2. Immunohistochemistry analysis suggests that cell therapy would promote a reconstructive response. Histology and HRCT analysis suggest a positive application of swine as a model for a bleomicine inducing of fibrotic interstitial pulmonary disease.

rst transcriptional activity influences kirre mRNA concentration in the Drosophila pupal retina during the final steps of ommatidial patterning.

BACKGROUND: Drosophila retinal architecture is laid down between 24-48 hours after puparium formation, when some of the still uncommitted interommatidial cells (IOCs) are recruited to become secondary and tertiary pigment cells while the remaining ones undergo apoptosis. This choice between survival and death requires the product of the roughest (rst) gene, an immunoglobulin superfamily transmembrane glycoprotein involved in a wide range of developmental processes. Both temporal misexpression of Rst and truncation of the protein intracytoplasmic domain, lead to severe defects in which IOCs either remain mostly undifferentiated and die late and erratically or, instead, differentiate into extra pigment cells. Intriguingly, mutants not expressing wild type protein often have normal or very mild rough eyes. METHODOLOGY/PRINCIPAL FINDINGS: By using quantitative real time PCR to examine rst transcriptional dynamics in the pupal retina, both in wild type and mutant alleles we showed that tightly regulated temporal changes in rst transcriptional rate underlie its proper function during the final steps of eye patterning. Furthermore we demonstrated that the unexpected wild type eye phenotype of mutants with low or no rst expression correlates with an upregulation in the mRNA levels of the rst paralogue kin-of-irre (kirre), which seems able to substitute for rst function in this process, similarly to their role in myoblast fusion. This compensatory upregulation of kirre mRNA levels could be directly induced in wild type pupa upon RNAi-mediated silencing of rst, indicating that expression of both genes is also coordinately regulated in physiological conditions. CONCLUSIONS/SIGNIFICANCE: These findings suggest a general mechanism by which rst and kirre expression could be fine tuned to optimize their redundant roles during development and provide a clearer picture of how the specification of survival and apoptotic fates by differential cell adhesion during the final steps of retinal morphogenesis in insects are controlled at the transcriptional level.

Analysis of GSTM1 and GSTT1 polymorphisms in circulating plasma DNA of lung cancer patients.

The concentration of free circulating plasma DNA and the genetic profile of patients suffering from various types of tumors were studied in an effort to increase the understanding of the biomarkers and genetic factors involved in predisposing an individual to lung cancer (LC). The polymorphic inheritance of glutathione S-transferases (GST), which modulate the effects of various genotoxic agents, especially those derived from benzo[a]pyrene, one of the main tobacco carcinogens, has been implicated in both cancer risk and prognostics. We investigated gene polymorphisms in the blood serum of patients previously diagnosed at the Pneumology Division of the Clementino Fraga Filho University Hospital of the Federal University of Rio de Janeiro and in buccal swab samples of exfoliated oral cells obtained from a population of healthy controls. The distribution of GSTM1 and GSTT1 polymorphisms was not significantly different between LC patients and the controls, suggesting that GSTM1 and GSTT1 alone or in combination are not independent risk factors for LC. However, a close relationship between smoking status and LC was clearly demonstrated. The most significant risk for LC concerning tobacco smoking was found in the association of null genotypes for GSTM1 and GSTT1 (P < 0.0001).

Morphological aspects of buffaloes (Bubalus bubalis) umbilical cord / Aspectos Morfológicos do cordão umbilical de búfalos (Bubalus bubalis)

Buffalo is an important livestock resource, with a great participation in agricultural systems, providing milk, meat, and work power. Umbilical cord is responsible for maternal-fetal nutrients exchange during pregnancy, and its alterations can compromise the fetal development. We investigated ten pregnant uteruses collected from cross-bread buffaloes in different stages of gestation. Pregnancy and fetal age was determined by measuring the apex sacral length and development period was calculated by previously published formula. Umbilical cords were measured for length determination. Umbilical cord vascular net and anastomosis were observed by injection of Neoprene latex. Histological sections of the umbilical cord were studied after stain with HE, picrossirius, toluidine blue, orceine, and PAS reaction. Buffaloes' umbilical cord was formed by two central arteries, an allantois duct and two peripheral veins. The artery wall was composed by large quantity of collagen, elastic fibers, fibroblasts and large number of vasa vasorum. The allantois duct was located between the arteries and presented a great number of small nourishing vessels. Small nourishing vessels should be carefully considered to avoid to be mistaken to the arterials and veins vasa vasorum. Medium length of umbilical cord from buffalos was 11.8cm (minimum of 6.8cm and maximum of 17.4cm).

Búfalo é uma importante fonte de recurso nos rebanhos animais, apresentando uma grande participação na agropecuária, provendo leite, carne e força de trabalho. O Cordão umbilical é responsável pela troca de nutrientes materno-fetais durante a gestação, e suas alterações podem comprometer o desenvolvimento fetal. Nós investigamos dez úteros gravídicos de búfalos de raças cruzadas em fases diferentes de gestação. O período de gestação e a idade fetal foram determinados pelo comprimento ápice sacral, aplicando fórmulas previamente estabelecidas. Posteriormente mediu-se o comprimento do cordão umbilical. A rede vascular do cordão umbilical e anastomoses foram observadas por injeção ou látex de neoprene. O cordão umbilical foi estudado a partir de cortes histológicos, corados por HE, picrossirius, azul de Toluidina, orceína e reação histoquímica de PAS. O cordão umbilical de búfalos é formado por duas artérias centrais, ducto alantóide e duas veias periféricas e apresentam forma de ampulheta. A parede da artéria umbilical é composta por grande quantidade de fibras colágenas e elásticas, fibroblastos e um grande número de vasa vasorum. O ducto alantóide fica alocado entre as artérias e apresenta um grande número de pequenos vasos nutritivos. Os vasos nutritivos devem ser cuidadosamente identificados para evitar-se confundi-los com vasa vasorum. O comprimento médio do cabo de cordão umbilical dos búfalos era 11.8cm (mínimo de 6.8cm e máximo de 17.4cm).

Chorion biopsy in mongrel dogs / Biopsia do cório fetal em cães

With the great development of the gestational studies in all of the species, we noticed the necessity of adaptations of these techniques for prenatal diagnosis in dogs. Based on this, we studied the feasibility of chorion biopsy guided by ultrasound. Our results demonstrated accuracy on the sex determination being 2 males and 12 females, as well as it would be possible to identify chromosome alteration due to the quality of samplings. Sex determination was accomplished with the identification of Y gene chromosomes in PCR technique. After the collection, fragments were prepared for light microscopy studies and revealed fetal chorion tissue, blood colloid and erythrocyte. In the whole material we found hemosiderin impregnations due to the hemolysis and to the residue of blood of the placental marginal hematomes. The submitted female dogs to this technique demonstrated normal puppy births without death.

Com o grande desenvolvimento dos estudos gestacionais em todas as espécies, percebemos a necessidade de adaptarmos técnicas para diagnóstico pré-natal para cães. Assim, buscamos bases nas técnicas já existentes empregadas em humanos, e através destas, conseguimos estabelecer um método para coleta em cães, utilizando PCR para garantirmos a integridade das amostras. O procedimento foi realizado através de punção da cinta placentária com agulha de biopsia guiada por ultra-som. De todas as 14 amostras coletadas, duas apresentaram-se positivas para o cromossomo Y, presente apenas em machos, confirmando assim a viabilidade das amostras demonstrando com isso que através desta técnica podemos coletar material fetal para diagnóstico de alterações gênicas ou cromossômicas presentes nos cães antes mesmo destes virem a termo. A microscopia de material revelou fragmentos de cório fetal, colóide sangüíneo e eritrócitos. Em todo o material encontramos impregnações de hemosiderina devido à hemólise e ao resíduo de sangue dos hematomas marginais placentários. As cadelas submetidas a esta técnica tiveram partos normais sem óbito de nenhum filhote.