Ectopic development of skeletal muscle induced by subcutaneous transplant of rat satellite cells

The present study analyzes the ectopic development of the rat skeletal muscle originated from transplanted satellite cells. Satellite cells (10(6) cells) obtained from hindlimb muscles of newborn female 2BAW Wistar rats were injected subcutaneously into the dorsal area of adult male rats. After 3, 7, and 14 days, the transplanted tissues (N = 4-5) were processed for histochemical analysis of peripheral nerves, inactive X-chromosome and acetylcholinesterase. Nicotinic acetylcholine receptors (nAChRs) were also labeled with tetramethylrhodamine-labeled alpha-bungarotoxin. The development of ectopic muscles was successful in 86% of the implantation sites. By day 3, the transplanted cells were organized as multinucleated fibers containing multiple clusters of nAChRs (N = 2-4), resembling those from non-innervated cultured skeletal muscle fibers. After 7 days, the transplanted cells appeared as a highly vascularized tissue formed by bundles of fibers containing peripheral nuclei. The presence of X chromatin body indicated that subcutaneously developed fibers originated from female donor satellite cells. Differently from the extensor digitorum longus muscle of adult male rat (87.9 ± 1.0 µm; N = 213), the diameter of ectopic fibers (59.1 µm; N = 213) did not obey a Gaussian distribution and had a higher coefficient of variation. After 7 and 14 days, the organization of the nAChR clusters was similar to that of clusters from adult innervated extensor digitorum longus muscle. These findings indicate the histocompatibility of rats from 2BAW colony and that satellite cells transplanted into the subcutaneous space of adult animals are able to develop and fuse to form differentiated skeletal muscle fibers.

Genomic analysis of Brazilian patients with Fabry disease

Fabry disease is an X-linked lysosomal disorder due to a-galactosidase A deficiency that causes storage of globotriaosylceramide. The gene coding for this lysosomal enzyme is located on the long arm of the X chromosome, in region Xq21.33-Xq22. Disease progression leads to vascular disease secondary to involvement of kidney, heart and the central nervous system. Detection of female carriers based solely on enzyme assays is often inconclusive. Therefore, mutation analysis is a valuable tool for diagnosis and genetic counseling. Many mutations of the a-galactosidase A gene have been reported with high genetic heterogeneity, being most mutations private found in only one family. The disease is panethnic, and estimates of incidence range from about 1 in 40,000 to 60,000 males. Our objective was to describe the analysis of 6 male and 7 female individuals belonging to 4 different Fabry disease families by automated sequencing of the seven exons of the a-galactosidase gene. Sequencing was performed using PCR fragments for each exon amplified from DNA extracted from peripheral blood. Three known mutations and one previously described in another Brazilian family were detected. Of 7 female relatives studied, 4 were carriers. Although the present study confirms the heterogeneity of mutations in Fabry disease, the finding of the same mutation previously detected in another Fabry family from our region raises the possibility of some founder effect, or genetic drift. Finally, the present study highlights the importance of molecular analysis for carrier detection and genetic counseling.

Bradysia hygida (Diptera, Sciaridae) presents two eukaryotic Elongation Factor 1A gene homologues: partial characterization of the eukaryotic Elongation Factor 1A-F1 gene

Elongation factor 1A is a highly conserved protein that participates in translation. We report the occurrence of two genes homologous to the eukaryotic Elongation Factor 1A in Bradysia hygida and describe the partial cloning and characterization of the B. hygida eukaryotic Elongation Factor 1A-F1 (BheEF1A-F1) gene. The pattern of BheEF1A-F1 expression in the salivary gland at the end of the fourth larval instar was investigated using real-time PCR. The results showed that BheEF1A-F1 expression levels are relatively constant at the time when rapid changes in protein synthesis occur in this tissue. In situ hybridization experiments coupled to Southern blot analyses showed that the BheEF1A-F1 gene is located at position 3d of the A chromosome and a second gene homologous to eEF1A is located at position 6a of the X chromosome. Southern blot analyses showed that both the BheEF1A-F1 gene and the second gene homologous to eEF1A constitute non-amplified genes. The present results contribute to the molecular characterization of a sciarid eEF1A gene.

Polytene chromosome map and inversion polymorphism in Drosophila mediopunctata

Drosophila mediopunctata belongs to the tripunctata group, and is one of the commonest Drosophila species collected in some places in Brazil, especially in the winter. A standard map of the polytene chromosomes is presented. The breakpoints of the naturally occurring chromosomal rearrangements are marked on the map. The distribution of breaking points through the chromosomes of D. mediopunctata is apparently non-random. Chromosomes X, II and IV show inversion polymorphisms. Chromosome II is the most polymorphic, with 17 inversions, 8 inversions in the distal region and 9 in the proximal region. Chromosome X has four different gene arrangements, while chromosome IV has only two.

Chromosome studies in some Stevia. Cav. (Compositae) species from Southern Brazil

Karyotypes of six species of the genus Stevia from Southern Brazil were studied, utilizing root tip metaphases. All species were diploid with 2n = 22 chromosomes. It was possible to identify each species by chromosome morphology. The basic chromosome number for Brazilian species of Stevia is X = 11. This number is also found in almost all South American species. We suggest that in Stevia there is an evolutionary trend toward chromosomal rearrangement, caused mainly by pericentric inversions. It was found that, in addition to aneuploidy and polyploidy, chromosomal rearrangements are common in the tribe Eupatorieae.

TP53 mutations and loss of heterozygosity of chromosome 17 in colorectal tumors

The incidence of TP53 point mutations and loss of heterozygosity (LOH) of chromosome 17 in colorectal tumors was determined in a group of Brazilian patients. We screened DNA samples from tumors and distal normal mucosa of 39 patients with colorectal cancer, for TP53 mutations by PCR-SSCP (single-strand conformation polymorphism) analysis. Chromosome 17 LOH was investigated using six PCR-based polymorphic markers and one VNTR probe. TP53 mutations were demonstrated in 15/39 of the cases. Mutations were distributed among all exons examined (five to eight), the majority of them being G/C to A/T transitions. LOH of chromosome 17p and 17q was detected in 70 and 46% of the tumors, respectively. There was a significant association between TP53 mutations and LOH in chromosome 17p (P = 0.0035) and 17q (P = 0.03). Although no correlation was observed between TP53 genetic alterations and clinical/ pathological characteristics, the association of TP53 mutations with loss of both chromosome 17 arms may indicate that TP53 inactivation provokes an unstable phenotype in tumor cells in colorectal tumors.

Study of a region on yeast chromosome XIII that complements pet G199 mutants (COX7) and carries a new non-essential gene

The mutants of Saccharomyces cerevisiae assigned to complementation group G199 are deficient in mitochondrial respiration and lack a functional cytochrome oxidase complex. Recombinant plasmids capable of restoring respiration were cloned by transformation of mutants of this group with a yeast genomic library. Sequencing indicated that a 2.1-kb subclone encompasses the very end (last 11 amino acids) of the PET111 gene, the COX7 gene and a new gene (YMR255W) of unknown function that potentially codes for a polypeptide of 188 amino acids (about 21.5 kDa) without significant homology to any known protein. We have shown that the respiratory defect corresponding to group G199 is complemented by plasmids carrying only the COX7 gene. The gene YMR255W was inactivated by one-step gene replacement and the disrupted strain was viable and unaffected in its ability to grow in a variety of different test media such as minimal or complete media using eight distinct carbon sources at three pH values and temperatures. Inactivation of this gene also did not affect mating or sporulation

Simultaneous allergen inactivation and detoxification of castor bean cake by treatment with calcium compounds

Ricinus communis L. is of great economic importance due to the oil extracted from its seeds. Castor oil has been used for pharmaceutical and industrial applications, as a lubricant or coating agent, as a component of plastic products, as a fungicide or in the synthesis of biodiesel fuels. After oil extraction, a castor cake with a large amount of protein is obtained. However, this by-product cannot be used as animal feed due to the presence of toxic (ricin) and allergenic (2S albumin) proteins. Here, we propose two processes for detoxification and allergen inactivation of the castor cake. In addition, we establish a biological test to detect ricin and validate these detoxification processes. In this test, Vero cells were treated with ricin, and cell death was assessed by cell counting and measurement of lactate dehydrogenase activity. The limit of detection of the Vero cell assay was 10 ng/mL using a concentration of 1.6 x 10(5) cells/well. Solid-state fermentation (SSF) and treatment with calcium compounds were used as cake detoxification processes. For SSF, Aspergillus niger was grown using a castor cake as a substrate, and this cake was analyzed after 24, 48, 72, and 96 h of SSF. Ricin was eliminated after 24 h of SSF treatment. The cake was treated with 4 or 8% Ca(OH)2 or CaO, and both the toxicity and the allergenic properties were entirely abolished. A by-product free of toxicity and allergens was obtained.

Fatores prognósticos no tratamento do carcinoma epidermóide da laringe: cirurgia conservadora x radical

O tratamento do carcinoma da laringe tem como desafio a cura do paciente e a preservação do órgão. OBJETIVO: A avaliação dos subsítios na disseminação do carcinoma e a sua influência no tratamento cirúrgico da laringe, no controle local e na sobrevida. MATERIAL E MÉTODO: Cento e sessenta pacientes tratados na Escola Paulista de Medicina - Hospital São Paulo, no período de janeiro de 1998 a dezembro de 2004, foram compilados para análise da disseminação do carcinoma dos subsítios da laringe. Foram incluídos pacientes com seguimento mínimo de 2 anos, após diagnóstico e tratamento. A avaliação estatística foi: Teste X2, Teste de Fisher e a Curva de Kaplan-Meier. Forma de Estudo: Estudo de Coorte Histórica Longitudinal (Trabalho Clínico Retrospectivo). RESULTADOS: A comissura posterior e a infraglote, respectivamente, foram significantes para a laringectomia total: tumores glóticos AC: (p=0,03) AP: (p=0,0001); AC: (p=0,0007) AP: (p<0,0001). A infraglote foi significante nos tumores G+SG na AP: (p=0,04), e na taxa de óbitos AP: (p=0,03). CONCLUSÃO: A laringectomia total é o tratamento de eleição na presença de alto comprometimento da comissura posterior e da infraglote. Esta última pode comprometer a sobrevida, conforme a invasão local, mesmo na presença de margens cirúrgicas livres.

O orçamento de defesa dos EUA: racionalidade X pressões domésticas

Ao longo das últimas décadas, gastos substanciais com defesa têm sido mantidos pelos EUA. Essas despesas simbolizam uma reação do país a condições sistêmicas, mas também se relacionam com os interesses de agentes domésticos que trabalham para sua manutenção. Neste artigo, pretende-se examinar como funcionou a combinação dessas pressões no período que marcou o fim da Guerra-Fria, por meio da comparação entre os orçamentos de defesa da primeira gestão de Reagan (1981-1984) e da Administração Bush (1989-1992).

Influência do tipo de parto e do sexo no desempenho e nas características da carcaça de cordeiros cruza Ile de France x Texel

O experimento foi realizado no Setor de Ovinocultura do Departamento de Zootecnia da Universidade Federal de Santa Maria (UFSM), com o objetivo de avaliar o efeito do tipo de parto e do sexo sobre o desempenho e as características da carcaça de cordeiros cruza Ile de France x Texel. Foram utilizados 12 cordeiros não castrados e seis cordeiras. O delineamento experimental adotado foi o inteiramente casualizado, com seis repetições. Os tratamentos foram: cordeiros de parto simples, cordeiros de parto duplo e cordeiras de parto duplo. O ganho de peso médio diário do nascimento aos 21 dias dos cordeiros de parto simples foi superior (P < 0,05) em relação aos cordeiros e cordeiras de parto duplo. Entretanto, após a fase inicial de amamentação, em que o leite deixa de ser o principal componente da dieta, verificou-se que os cordeiros de parto simples apresentaram desempenho semelhante ao dos de parto duplo. Os cordeiros tiveram ganho de peso superior (P < 0,05) aos das cordeiras após os 42 dias de vida. As características da carcaça de cordeiros de parto simples e cordeiros e cordeiras de parto duplo são similares, com exceção do peso de carcaças quente e fria e do comprimento interno de carcaça e perna, em que o dos machos são superiores ao das fêmeas.

Regionalização para o cultivo do feijão no Rio Grande do Sul com base na interação genótipo x ambiente

A regionalização de uma cultura agrícola é estratégia utilizada para minimizar efeitos da interação genótipo x ambiente. Esta abordagem baseia-se em resultados de experimentos conduzidos em diversos locais e anos, cuja análise estatística apresenta dificuldades originadas de estrutura de dados incompleta e heterogeneidade da variância do erro experimental. Este estudo teve o objetivo de avaliar a magnitude e a natureza da interação genótipo x ambiente e a possibilidade de estratificação das regiões de cultivo do feijão do Estado do Rio Grande do Sul. Foram considerados dados do Ensaio Estadual de Feijão, conduzido em 24 locais durante oito anos. Foram procedidas análises conjuntas, para cada ano, de dois subconjuntos de quatro anos e do conjunto dos oito anos, e análises de agrupamento, utilizando metodologia que contorna aquelas dificuldades da análise estatística. Os resultados das análises conjuntas anuais revelaram alta significância da interação genótipo x local. Essa interação também foi significativa no conjunto dos oito anos, mas não significativa nos dois subconjuntos de quatro anos. A interação genótipo x local x ano foi altamente significativa nesses três subconjuntos de anos. A análise de agrupamento, baseada nos oito anos, constituiu subregiões incoerentes com as subregiões formadas pelas análises anuais, que também foram inconsistentes entre si. Esses resultados indicaram inadequabilidade da regionalização do Estado do Rio Grande do Sul.

Uso de fitorreguladores no desenvolvimento de frutos na atemoieira (Annona cherimola x A. squamosa cv. Gefner)

Este trabalho foi realizado com atemoieiras 'Gefner', com o objetivo de avaliar o efeito de fitorreguladores no pegamento de frutos. Foram realizados três experimentos, no município de Matias Cardoso, Minas Gerais. No primeiro, avaliaram-se quatro tipos de auxinas: os ácidos indol butírico (AIB), indol acético (AIA), naftaleno acético (ANA) e diclorofenoxiacético (2,4-D), combinados com três concentrações (150, 300 e 450 mg L-1). O ácido giberélico (GA3) também foi aplicado, na dosagem de 1 g L-1, aos 14 e 21 dias após a primeira aplicação das auxinas. No segundo experimento, testaram-se o AIA, AIB e ANA, combinados em duas concentrações (450 e 600 mg L-1). No terceiro experimento, avaliou-se a aplicação do ANA na dose de 450 mg L-1 associado com número de aplicações: semanal até 35 dias; semanal até 70 dias; semanal até 105 dias e a cada duas semanas. O GA3 foi utilizado na dosagem de 1 g L-1, em quatro aplicações, aos 13, 27, 41 e 54 dias após a primeira aplicação do ANA. Em todos os experimentos, a testemunha foram flores polinizadas artificialmente. ANA foi a auxina sintética que propiciou as maiores médias de pegamento dos frutos, com 8% até aos 148 dias após a primeira aplicação. A utilização de 2,4-D acarretou as menores percentagens de pegamento dos frutos; no 14° dia da aplicação, mais nenhum fruto foi observado. Independentemente do tipo de fitorregulador, os tratamentos não permitiram o desenvolvimento de frutos comerciais de atemoia.