Pancreatic adenocarcinoma is important in oncology because of its high mortality rate. Deaths may be avoided if an early diagnosis could be achieved. Several types of tumors overexpress gastrin-releasing peptide receptors (GRPr), including pancreatic cancer cells. Thus, a radiolabeled peptide derivative of gastrin-releasing peptide (GRP) may be useful as a specific imaging probe. The purpose of the present study was to evaluate the feasibility of using99mTc-HYNIC-βAla-Bombesin(7-14)as an imaging probe for Capan-1 pancreatic adenocarcinoma. Xenographic pancreatic tumor was developed in nude mice and characterized by histopathological analysis. Biodistribution studies and scintigraphic images were carried out in tumor-bearing nude mice. The two methods showed higher uptake by pancreatic tumor when compared to muscle (used as control), and the tumor-to-muscle ratio indicated that99mTc-HYNIC-βAla-Bombesin(7-14)uptake was four-fold higher in tumor cells than in other tissues. Scintigraphic images also showed a clear signal at the tumor site. The present data indicate that99mTc-HYNIC-βAla-Bombesin(7-14)may be useful for the detection of pancreatic adenocarcinoma.

Para avaliar a viabilidade da metodologia da Reação em Cadeia da Polimerase associada com o Polimorfismo de Fragmentos de DNA (PCR-RFLP) na identificação de fraude intencional e contaminação acidental em produtos cárneos de origem bubalina, in natura e processados, foram testadas amostras puras e amostras de carnes com misturas controladas, produzidas em laboratório, com adição de 1%, 5%, 10% e 50% de carne bovina em carne de búfalo, homogeneizada crua e em amostras autoclavada. Foram comparados, ainda, diferentes métodos de extração, usando um kit comercial e a técnica clássica, utilizando fenol/clorofórmio. O resultado estatístico foi obtido por tabela de contingência, analisada pelo teste do qui-quadrado (χ2) e do exato de Fisher. A especificidade encontrada foi altamente significativa (P<0,0001). Observou-se também sensibilidade altamente significativa nas diluições a partir de 10% (P<0,0001). A técnica tem alta especificidade e sensibilidade para detectar até mesmo contaminação de 1%, mas a repetibilidade desse resultado impede a aplicação oficial desse método para a inspeção de contaminação acidental, sendo recomendada somente para inspeção de fraude a partir de 10% de substituição. Em carnes autoclavadas, a eficácia do teste é menor. A técnica pode ser empregada para certificação de produto específico (selo de identidade de espécie).

The present study aimed at evaluate the viability of PCR-RFLP (Polymerase Chain Reaction - Restriction Fragment Length Polymorphism) for identification of fraud and/or accidental contamination in buffalo meat - fresh and processed. Pure, autoclaved and controlled fraud samples, produced in the laboratory with the addition of 1, 5, 10 and 50% of beef in raw homogenized buffalo meat samples, were tested. Furthermore, different extraction methods, using a commercial kit and classical technique using phenol-chloroform, were compared. The statistical result was obtained by contingency table analyzed by chi-square and the Fisher exact test. The specificity was highly significant (p <0.0001), and the sensitivity was highly significant in dilutions from 10% (p <0.0001). Despite its accuracy and precision, capable to detect a contamination level of 1%, PCR-RFLP technique is not recommended for inspection in cases of accidental contamination. This is due to the need of test repetition in levels of contamination lower than 10%. The efficiency of this test is lower to autoclaved meat. The PCR-RFPL technique can be used for certification of food made with specific species (species identification certification stamp).

Foram testados três métodos de extração de DNA em amostras de queijo, com o objetivo de identificar uma técnica eficiente para extração de DNA em amostras com várias limitações, como alto teor de gordura, alto grau de degradação do DNA e grande concentração de impurezas. A técnica que faz uso do tiocianato de guanidina mostrou-se mais adequada para identificação de adição intencional não declarada de leite bovino em queijos bubalinos, podendo ser empregada para certificação de produto específico (selo de Identidade de Espécie).

Three methods of DNA extraction were tested in cheese samples. The objective of this study was to identify an efficient technique for DNA extraction in different samples with several limitations, such as high fat tenor, high degree of DNA degradation and great sludge concentration. The technique using Guanidine thiocyanate was more appropriate for identification of intentional undeclared addition of bovine milk in buffalo cheeses. This technique can be used for certification of a specific product (stamp of Identity of Species).

Foram utilizados 159 cavalos Pampa, registrados na Associação Brasileira dos Criadores de Cavalo Pampa, e um grupo-controle, de 32 cavalos da raça Paint, ambos os grupos provenientes de plantéis de diferentes regiões brasileiras, com o objetivo de comparar os testes bioquímico e molecular para detecção de marcadores genéticos para pelagem tobiana em cavalos Pampa. Houve diferença significativa (P<0,001) entre os testes bioquímico e molecular, nos cavalos Pampa, mas o mesmo fato não ocorreu com os da raça Paint. Os resultados mostraram que o marcador molecular (KIT) foi mais eficiente na identificação dos prováveis cavalos homozigotos do que os marcadores bioquímicos albumina (Al) e proteína de ligação da vitamina D (Gc), em ambas as raças.

In this study, 159 Pampa horses, registered at the Associação Brasileira dos Criadores de Cavalo Pampa, and a control group of 32 Paint horses, both coming from herds located in different Brazilian regions, were used to compare biochemical and molecular tests for detection of genetic markers for the Tobiano coat color pattern in Pampa horses. Difference (P<0.001) between biochemical and molecular tess in Pampa horses was observed, but not for the Paint horses. The results showed that the molecular marker (KIT) was more efficient to identify the probable homozygous dominant horses than the biochemical markers albumin (Al) and vitamin D-binding Protein (Gc), in both breeds.

Utilizaram-se 195 cavalos Pampa e um grupo-controle de 41 cavalos da raça Paint, provenientes de plantéis de várias regiões brasileiras, com o objetivo de avaliar a eficiência do teste mediante uso de marcadores bioquímicos: albumina (Al) e proteína de ligação da vitamina D (Gc), para identificação dos possíveis indivíduos homozigotos dominantes para o padrão de pelagem tobiano nos cavalos Pampa. Não foram encontrados genótipos AlBB e GcSS, revelando indício de quebra de ligação gênica entre tais locos e o loco tobiano e a ineficácia do teste bioquímico na detecção dos prováveis indivíduos homozigotos dominantes para o padrão de pelagem tobiano nos cavalos Pampa.

One hundred and ninety five Pampa horses and a control group of 41 Paint Horses originated from herds located in different Brazilian regions were used to objective of evaluate the efficiency of the biochemical markers albumin (Al) and vitamin D binding protein (Gc) to identify the probable homozygous dominant for the tobiano coat color pattern in Pampa horses. It was not found any genotype AlBB and GcSS, indicating a possible break of the genetic linkage between these loci and the locus Tobiano, as well as the inefficacy of the biochemical test in the detection of the probable homozygous dominant for the tobiano color pattern in Pampa horses.

Identificou-se e determinou-se a freqüência do gene miostatina (GDF-8) normal e mutante em rebanhos Marchigiana, em 377 bovinos da raça Marchigiana, criados nos estados de São Paulo e Paraná. Identificaram-se 37,9% de animais normais, 55,2% de portadores e 6,9% homozigotos afetados para musculatura dupla. Estes resultados indicam que os criadores têm interesse na característica musculatura dupla, promovendo, ainda que aleatoriamente, selecão a favor da mutação.

The frequency of the normal myostatin gene (GDF-8) and the mutant allele in Marchigiana herds was detected. Three hundred and seventy-seven animals of Marchigiana breed raised in São Paulo and Paraná States, Brazil, were tested. The results showed that 37.9% were homozygous normal animals, 55.2% heterozygous and 6.9% homozygous double muscling. The results suggest the interest of the breeders in having interest in the character double muscling, randomly promoting, the selection in favor of the mutation.

Foram caracterizados geneticamente, utilizando-se cinco locos de microssatélites, 235 indivíduos de seis plantéis de tilápia (Ceará, Chitralada, Israel, Nilótica, Taiwan e Vermelha) da região Sudeste do Brasil. Verificou-se diferença genética entre os seis plantéis, obtida pelo cálculo do índice de fixação de alelos (Fst=0,3263). De modo geral, está ocorrendo perda de heterozigose nos plantéis, segundo mostrou a estimativa do coeficiente de endogamia intrapopulacional (Fis=0,0486). Os plantéis Israel e Nilótica foram os mais semelhantes geneticamente (Ig=0,6663). Os plantéis Chitralada e Taiwan foram os que menos apresentaram genes em comum (Ig=0,2463). O plantel denominado Vermelha foi o mais distinto entre todos.

Two hundred and thirty five individuals from six commercial stocks of tilapias (Ceará, Chitralada, Israel, Nilótica, Taiwan and Red) from the Southeastern region of the country were genetically characterized using five microsatellite loci. The results suggest large genetic difference among the stocks, estimated through the fixation allele index (Fst = 0.3263), and a considerable loss of heterozigosity accurs in most of the stocks, according to the population inbreeding coefficient (Fis=0.0486). The Israel and Nilótica stocks were genetically similar (Ig=0.6663), while Chitralada and Taiwan showed less genes in common (Ig=0.2463). The Red stock was the most distinct stock.

DNA samples of six bovines obtained from three tissues (blood, semen and hair) were extracted using two different techniques. After the extraction procedures the samples were divided in six fractions. Three were stored at -20° C and three at 4° C. Every three months one sample of each tissue/extraction procedure was analyzed in spectrophotometer, to determine the quantity of the DNA and the extract was amplified using the primer RM 29. No differences in the DNA quantity or in the level of protein contamination among the three periods of analyses were observed. All the DNA extracted by quick extraction technique showed good amplification patterns during the nine months, meaning that this technique can be used in laboratory routine instead of the permanent extraction technique. The extract obtained from blood, using the permanent extraction technique, showed the higher quantity of DNA with the smaller index of protein contamination. The high quantity of protein contamination found in the semen samples preserved in egg yolk demanded modifications in both extraction techniques. After that the results were positive, showing good amplification patterns.

To evaluate the precision of the DNA tests using the non-automatized technique for individual identification and parentage tests, 105 Rottweiler dogs were studied using the primer CMR S. The sample was composed of 39 animals belonging to 11 complete families and their progenies, and 66 non related individuals until the second generation, derived from kennels located in the states of Minas Gerais and São Paulo. The CMR S primer was used for the Polimerase Chain Reaction (PCR). The results showed the inefficiency of the technique, even when analyzed through the automated gel analysis system. Also showed the impossibility of its commercial use due to the fact of does not permit the storage of data for subsequent use.

Foram utilizados 46 animais da raça Gir, registrados na Associação Brasileira de Criadores de Zebu, provenientes de cinco fazendas situadas no Estado de Minas Gerais, com o objetivo de avaliar a eficiência dos microssatélites BM2113, ILSTS005, ILSTS008, ETH131 e RM88 em testes de verificação de parentesco. Os locos BM2113, ILSTS005, ETH131 e RM88 mostraram-se eficientes, apresentando valores de PE2 (probabilidade de exclusão quando os dois progenitores são genotipados) entre 0,62 e 0,69 e PIC2 (conteúdo de informação polimórfica quando os dois progenitores são genotipados) entre 0,78 e 0,83. O mesmo não ocorreu para o loco ILSTS008, o qual apresentou baixos valores de PE2 (0,24) e PIC2 (0,41).

Forty six animals of the Gir breed, registered at the Brazilian Association of Zebu Breeders, coming from five farms located in Minas Gerais State, were used to analyze the efficiency of the microsatellites BM2113, ILSTS005, ILSTS008, ETH131 and RM88 in parentage tests. The loci BM2113, ILSTS005, ETH131 and RM88 showed to be efficient, presenting values of PE2 (exclusion probability when both parents are genotiped) between 0.62 and 0.69 and PIC2 (polymorphic information contents when both parents are genotiped) between 0.78 and 0.83. The same was not observed for the locus ILSTS008 that showed low values of PE2 (0.24) and PIC2 (0.41)

Neste estudo foram genotipados 205 cavalos da raça Árabe, criados nos estados de Minas Gerais e São Paulo, via DNA por meio de PCR, para determinação da presença do gene mutante SCID. Os resultados mostraram 98,5% de animais normais (202/205) e 1,5% de portadores (3/205). Pela análise da genealogia dos portadores identificados pode-se, ainda, confirmar a participação de um garanhão anteriormente identificado como provável disseminador da doença.

In the present study, 205 Arabian horses raised in Minas Gerais and São Paulo states, Brazil, were genotyped using the PCR technique. The results showed 98.5% of normals (202/205) and 1.5% of SCID carriers (3/205). Checking the pedigrees of the three carriers was possible to confirm the participation of one stallion first identified as the possible disease disseminator.