Relation between mast cells concentration and serotonin expression in chagasic megacolon development.

Chagas' disease is still reaching about 10 million people in the world. In South America, one of the most severe forms of this disease is the megacolon, characterized by severe constipation, dilated sigmoid colon and rectum and severe malnutrition. Previous data suggested that mast cells and serotonin (5-hydroxytryptamine [5-HT]) expression could be involved in intestinal homeostasis control, avoiding the chagasic megacolon development. The aim at this study was to characterize the presence of mast cells and expression of serotonin in chagasic patients with and without megacolon and evaluate the relation between mast cells, serotonin and megacolon development. Our results demonstrated that patients without megacolon feature a large amount of serotonin and few mast cells, while patients with megacolon feature low serotonin expression and a lot of mast cells. We believe that serotonin may be involved in the inflammatory process control, triggered by mast cells, and the presence of this substance in large quantities of the intestine could represent a mechanism of megacolon prevention.

Association between IGF2 and CYP21 gene polymorphisms and characteristics of economic interest in Nellore cattle.

We analyzed two single nucleotide polymorphisms (SNPs) of the IGF2 and CYP21 genes in Nellore cattle participating in the Brazilian Animal Breeding Program. The SNPs were found in exon 6 of the IGF2 (insulin-like growth factor 2) gene (RFLP/MboII) as well as in the promoter region of the CYP21 (steroid 21-hydroxylase) gene (RFLP/HpaII) of these animals. The TC heterozygotes were significantly more frequent than CC and TT homozygotes in the RFLP/MboII polymorphism. The T allele was significantly more frequent than the C allele in RFLP/HpaII polymorphism. This population was found to be in Hardy-Weinberg equilibrium for these SNPs. Association of these polymorphisms with expected progeny differences of reproductive and productive traits was investigated, but proved to be significant only for DP550 (expected progeny differenced for weight at 365 days - IGF2 - RFLP/MboII) and DP450 (expected progeny differenced for weight at 450 days - CYP21 - RFLP/HpaII). This is the first study on the occurrence of these two polymorphisms in this Zebu breed of cattle. A total of 147 Nellore animals participating in the Breeding Program of the Nellore Breed (PMGRN) under the management of the National Association of Breeders and Researchers (ANCP) in the city of Ribeirão Preto were analyzed.

Identification of a DNA methylation point in the promoter region of the bovine CYP21 gene.

The CYP21 (steroid 21-hydroxylase) gene is involved in the synthesis of steroid hormones. Bov-A2 is a retroposon that is common in ruminant genomes. The promoter region of bovine CYP21 contains a short interspersed nucleotide element of Bov-A2, which overlaps a putative Sp1 binding site. We looked for RFLP/HpaII polymorphism in the Bov-A2 element in bovine Zebu breeds by PCR-RFLP, and examined whether polymorphism in this element is associated with methylation. Among DNA samples from 135 Brazilian Zebu breed cattle, we identified an RFLP/HpaII polymorphism (T/C), which, based on a restriction methylation-sensitive assay employing HpaII and isoschizomer MspI enzymes (methylation-sensitive and -non-sensitive enzymes, respectively), appears to be a DNA methylation point. This is the first report of this polymorphism and on DNA methylation in the bovine CYP21 promoter region in Brazilian Zebu cattle.

IGF2/Mboll polymorphism in Gir and Nelore cattle.

The imprinted gene insulin-like growth factor 2 (IGF2) carries out important functions in the development of placental mammals, during the embryonic and fetal stages and is located on bovine chromosome 29. The main aim of this study was to verify the occurrence of a polymorphism (C/T transversion) in exon 6 of the IGF2 gene in Gir and Nelore cattle, Zebu breeds of great economic importance in Brazil for dairy and beef production, respectively. A 193 bp fragment of bovine IGF2 exon 6 was amplified by PCR with specific primers for this region and digested with Mboll enzyme to analyze the polymorphic segment. In a total of 39 Gir animals (29 females and 10 males), the estimated frequencies of the C and T alleles were 0.42 and 0.58, respectively. The genotypic frequencies were 0.10 for CC, 0.26 for TT and 0.64 for CT. For 28 Nelore animals (17 females and 11 males), the estimated frequencies of the C and T alleles were 0.43 and 0.57, respectively. The genotypic frequencies were 0.18 for CC, 0.32 for TT, and 0.50 for CT. This is the first report of the occurrence of this polymorphism in these breeds. The polymorphic site can be a regulatory motif with functional significance to gene regulation of the IGF2 gene. This polymorphism could be used to investigate the allele-specific expression of the IGF2 gene, its epigenetic status, and its role in developmental, growth and reproductive traits.

Neuronal plasticity of the enteric nervous system is correlated with chagasic megacolon development.

Chagas' disease is one of the few functional gastrointestinal disorders for which a causative agent has been identified. However, some pathological aspects of the chagasic megasyndromes are still incompletely understood. Chagasic megacolon is characterized by an inflammatory process, organ dilatation and neuronal reduction in both plexuses of the enteric nervous system (ENS). Although some studies on the ENS in Chagas' disease have been performed, the process of neuronal destruction and neuronal regeneration still remains unclear. Our hypothesis is that the regeneration process of the ENS may be involved with the mechanisms that prevent or retard organ dilatation and chagasic megacolon development. For that reason, we evaluated the neuronal regeneration with the marker GAP-43 in the colon's neuronal plexuses from chagasic patients with megacolon, and from non-infected individuals. Visual examination and quantitative analysis revealed an increased neuronal regeneration process in the dilated portion from chagasic patients when compared with the non-dilated portion and with non-infected individuals. We believe that this increased regeneration can be interpreted as an accentuated neuronal plasticity that may be a response of the ENS to avoid megacolon propagation to the entire organ and maintain the colon functional innervation.

Alternativas para seleção de touros da raça Nelore considerando características múltiplas de importância econômica / Nellore bull selection alternatives using multiple traits of economic importance

Os dados são provenientes de 234 touros da raça Nelore participantes de um teste de progênie, no período de 1996 a 2003. A diferença esperada na progênie (DEP) de sete características: peso aos 120 e 210 dias, efeito materno (DMPP120 e DMPP210), peso e perímetro escrotal aos 365 e 450 dias, efeito direto (DDP365, DDP450, DDPE365 e DDPE450) e idade ao primeiro parto (DDIPP) foi utilizada para classificar os animais em três grupos, assim como identificar quais as características possuíram maior poder discriminatório na formação de cada grupo. Para tanto, foram utilizados procedimentos estatísticos multivariados de análise de agrupamentos k-médias e componentes principais. Os resultados evidenciaram que, dos três grupos formados, dois se destacaram quanto aos valores médios das DEPs. A importância desses dois grupos de touros foi confirmada pela análise de componentes principais, que associou a eles valores superiores de DEPs diretas de peso e perímetro escrotal. A quantidade da variabilidade original retida pelos dois primeiros componentes principais foi de 70,22 por cento. Estes procedimentos mostraram-se eficientes e constituíram importantes ferramentas para classificar touros, discriminar variáveis, bem como resumir informações multivariadas, podendo ser usados como auxílio valioso na seleção de reprodutores para uso nos programas de melhoramento genético.

The data set is from 234 Nelore bulls which participate of a progeny test in the period from 1996 to 2003. The expected progeny diference (EPDs) of seven economic traits, weights at 120 e 210 days of age, maternal effect (DMPP120 and DMPP210), weights and scrotal circumferences at 365 e 450 days of age, direct effects (DDP365, DDP450, DDPE365 and DDPE450), and age at first calving (DDIPP), were used in order to classify these animals in three groups and verify which EPDs showed greatest discriminating power in forming the groups. The statistical applied techniques were: k-means clusters analysis and principal components analysis. From the three groups formed, two of them stood out in relation to values of the EPDs means. Evidence of the importance of these two groups was observed in the principal component analysis that associate to them higher values of direct EPDs of weight and scrotal circumference. The two first principal components accounted for 70.22 percent of the total original variability. Both techniques could be an important tools to sire classification, variable discrimination, as well as to resume multivariate information, and they could be applied to help selection in animal breeding program.

Análise econômica de esquemas alternativos de controle leiteiro / Economic analysis of alternative schemes of milk recording

Compararam-se os custos de seis esquemas de controle leiteiro (CL), estabelecidos com base em diferentes freqüências de controles (com possibilidade de controle trimestral supervisionado e aproveitamento do CL da fazenda), no número de ordenhas e na realização de análises qualitativas (composição do leite e/ou contagem de células somáticas). Computaram-se os percentuais que os custos totais do CL representariam na receita bruta mensal do leite (CL/RB) e na receita líquida mensal (CL/RL). O esquema tradicional (freqüência mensal) apresentou o mais elevado custo mensal e os maiores CL/RB e CL/RL. Esquemas com intervalo maior entre controles e amostragem em ordenhas alternadas resultaram na redução dos custos com diárias e quilometragem. O custo mensal do CL variou de 0,68% a 1,8% sobre a receita bruta do leite e de 6,6% a 17,0% sobre a receita líquida.

A single step duplex PCR to distinguish Entamoeba histolytica from Entamoeba dispar.

In this study, a single-step duplex polymerase chain reaction procedure was developed for rapid, specific and sensitive identification of Entamoeba histolytica and for its diagnostic differentiation from E. dispar. Specific oligonucleotide primers were combined for the amplification of a cysteine proteinase 5 gene target sequence of 242 bp, present only in E. histolytica. Additionally, another oligonucleotide primer pair for both the E. histolytica and E. dispar actin gene target of 300 bp was designed to amplify only from amoebae DNA. The PCR developed was specific and efficiently identified and differentiated these parasites from each other in either cultured parasites or from stool material.

Indicadores de desempenho em rebanho da raça Holandesa: curvas de crescimento e altura, características reprodutivas, produtivas e parâmetros genéticos / Productive indicator in a Holstein dairy herd: growth and height curves, reproductive and productive traits and genetic parameters

Foram analisados dados de peso, altura e desempenho produtivo e reprodutivo de 1.272 novilhas da raça Holandesa, nascidas no período de 1990 a 1998. Estudaram-se o desenvolvimento em peso e altura do nascimento até o primeiro parto, as características peso ao primeiro parto (PP), idade ao primeiro parto (IP), produção de leite na primeira lactação (PL), primeiro intervalo de partos (IEP), e os parâmetros genéticos pertinentes. As equações para as curvas de peso e altura, estimadas pelo modelo proposto por von Bertalanffy, foram: Yi= 802,10 (1 - 0,630EXP (-0,0022t)) e Yi= 1,40 (1 - 0,208EXP (-0,0038t)), respectivamente. As novilhas atingiram o primeiro parto com altura média de 140cm e peso de 602kg aos 27 meses de idade. A produção total de leite na primeira lactação foi de 8.026kg e o primeiro IEP 420 dias. Na obtenção dos componentes de (co)variância utilizou-se o método da máxima verossimilhança restrita livre de derivada. As estimativas de herdabilidade foram 0,31, 0,35 e 0,34, respectivamente, para PP, IP e PL, e 0,10 para IEP, evidenciando variância genética aditiva média para as três primeiras características. As correlações genéticas apresentaram pequena magnitude para PP e PL, PL e IP, e IEP e PL. A correlação entre IEP e PP foi 0,49; entre IEP e IP o valor foi negativo, -0,69, indicando provável antagonismo entre as duas últimas características.

Genetic parameters of test day milk yields of Holstein cows

No presente estudo foram utilizados 17.968 registros de produçäo de leite, referentes a 2.130 primeiras lactaçöes de vacas da raça Holandesa, paridas nos anos de 1988 a 1991, filhas de 136 touros e controladas pela Associaçäo Brasileira de Criadores (ABC). Os dados foram distribuídos em dez sub-arquivos de acordo com o número do controle (M1 e M10). As características estudadas foram: produçäo de leite no dia do controle (M) e produçäo aos 305 dias de lactaçäo (M305), as quais foram ajustadas para os seguintes fatores de variaçäo: idade da vaca ao parto em classes, intervalo parto-primeiro controle e subclasses de rebanho-ano-estaçäo de parto. Os componentes de (co)variância foram obtidos a partir de análises com duas características utilizando-se o método REML, sob um modelo de touro. As estimativas de herdabilidade para as M variaram de 0,04 a 0,32 sendo os maiores valores encontrados na segunda fase da lactaçäo (M5 a M7), enquanto que para a M305 este valor foi de 0,32. As correlaçöes genéticas entre as M e a M305 variaram de 0,78 a 1,00. Os resultados sugerem que as M podem ser utilizadas em substituiçäo à M305 nas avaliaçöes genéticas dos animais, visto que as herdabilidades no meio da lactaçäo foram semelhantes à encontrada para a M305. Além disso, a seleçäo antecipada pode levar a uma reduçäo no intervalo de geraçöes.