Avaliação dos níveis séricos de triiodotironina (T3) e tiroxina (T4) durante o ciclo estral em marrãs (Sus scrofa domestica - Linnaeus, 1758)

No presente trabalho, foram utilizadas 21 fêmeas suínas, virgens, sexualmente aptas, criadas e mantidas sob condições industriais, para observação dos perfis hormonais séricos de triiodotironina e tiroxina, durante o ciclo estral. As colheitas de sangue foram efetuadas sempre no mesmo intervalo, entre 8 e 10 horas. Cada animal foi submetido a 14 punções venosas, distribuídas nos dias zero, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 21, 22 e 23 do ciclo estral. Consideramos o dia zero como o primeiro dia da fase estral, e o 23º dia como o primeiro do estro subseqüente. Os ensaios para dosagens hormonais foram executados utilizando-se a técnica de radioimunoensaio (RIE) em fase sólida e para isso foi empregado conjunto de reagentes comerciais (Coat-A-Count®). Para o hormônio triiodotironina foram observados valores altos no 1º e 23º dia e valor mínimo no 12º dia do ciclo estral. O hormônio tiroxina apresentou valores que oscilaram entre 2,42 e 3,74 mig/dl.

Avaliação dos níveis séricos de 17-alfa-OH progesterona e androstenediona durante o ciclo estral em marrãs (Sus Scrofa Domestica - Linnaeus, 1758)

No presente trabalho, foram utilizadas 21 fêmeas suínas, virgens, sexualmente aptas, criadas e mantidas sob condições industriais, para observação dos perfis hormonais séricos de 17-alfa-OH progesterona e androstenediona, durante o ciclo estral. As colheitas de sangue foram efetuadas sempre no mesmo intervalo, entre 8 e 10 horas. Cada animal foi submetido a 14 punções venosas, distribuídas nos dias zero, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 21, 22 e 23 do ciclo estral. Considerou-se o dia zero como o primeiro dia da fase estral, e o 23º dia como o primeiro do estro subseqüente. Os ensaios para dosagens hormonais foram executados utilizando-se a técnica de radioimunoensaio (RIE) em fase sólida e para isso foi empregado conjunto de reagentes comerciais (Coat-A-Count®). Para o hormônio 17-alfa-OH progesterona, foram encontrados valores médios que variaram entre 0,18 e 2,7 ng/ml e para o hormônio androstenediona esses valores oscilaram entre 0,08 e 0,24 ng/ml.

Evaluation of thyroxine (T4) and progesterone-17-alpha-OH circadian rhythm in swine females (Sus scrofa domestica - Linnaeus, 1758)

No presente trabalho foram utilizadas quatro fêmeas suínas, adultas, mestiças, não-gestantes e sem sinais clínicos de estro, criadas e mantidas sob condições industriais de criação. Objetivou-se avaliar a ocorrência de ritmicidade biológica circadiana para tiroxina e 17-alfa -OH progesterona. Os ensaios para dosagens hormonais foram executados utilizando-se a técnica de radioimunoensaio (RIE) em fase sólida e para isso foi empregado conjunto de reagentes comerciais (COAT-A-COUNT R). As análises séricas de tiroxina mostraram valores mais elevados ao redor das 15 horas, decrescendo a partir dai até atingir níveis menores no intervalo da zero às 4 horas. Quanto a 17-alfa -OH progesterona, observaram-se níveis mais elevados por volta das 3 horas, decrescendo gradativamente ao longo do dia, até atingir menor concentração no intervalo das 12 às 15 horas.

STRUCTURE OF THE RETE TESTIS OF THE CAT (FELIS-DOMESTICA, L)

The rete testis of the cat consists of 3 parts: a septal or interlobular part; a mediastinal part and a tunical part. The septal part contains the septal or transitory tubuli recti and the tubuli recti. The transitory tubules are formed as a confluence of the seminiferous tubules at the apex of the testicular lobules and the tubuli recti. The mediastinal rete is formed of long, straight channels which increase in size and become more irregular and anastomotic below the tunica albuginea at the cranial extremity of the testis. The end is characterized as the tunical part of the rete testis and communicates with the extratesticular rete testis. The channels all parts of the rete are lined by simple cuboidal or columnar epithelium. These epithelial channels are supported by a connective tissue containing smooth muscle cells. The framework tissue of the rete is more conspicuous at the cranial extremity of the testis, with a mio-connective matrix organization.

Níveis séricos de cortisol e 17-& estradiol durante o ciclo estral em marrãs (Sus domestica - Linnaeus 1758)

In the present study 21 young swine females, sexually mature, reared and kept under an industrial management system were assessed for cortisol and oestradiol-17 beta serum prolifes during the oestrus cycle. Blood sampling was performed always at the same interval, from 8:00 to 10:00 a.m. Each animal was submitted to 14 venal punctions at days 0, 2, 4, 6, 8, 10, 12, 14, 16, 18, 20, 21, 22 and 23 of the oestrous cycle. The first day of the oestrous phase was assumed to be day 0 and the 23(rd) as the first of the next cycle. Hormone determinations were performed using a solid phase radioimmunoassay (RIA). Regarding to cortisol concentrations, there was a variation from 3.5 to 8.0 mu g/dl and for oestradiol-17 beta the mean values detected varied from 3.5 to 14.9pg/ml.

Endophytic yeasts and filamentous fungi associated with southern Brazilian apple (Malus domestica) orchards subjected to conventional, integrated or organic cultivation

Endophytic fungi were isolated from leaves, flowers and fruit of healthy apple trees (Malus domestica, BORKH.) growing in southern Brazilian orchards under three different cultivation systems (conventional, integrated and organic), during two vegetative cycles. The greatest total number of endophytic isolates was obtained from the orchards under organic cultivation when compared to integrated and conventional cultivation systems. Filamentous fungi from the genera Colletotrichum, Xylaria and Botryosphaeria were the most frequent ones and the most representative yeast genera were Sporobolomyces, Rhodotorula, Debaryomyces and Cryptococcus. It is suggested that some isolates may be used as indicators of the different management systems. © 2005 Wiley-VCH Verlag GmbH & Co. KGaA.

Genetic diversity in wild (Sus scrofa scrofa) and domestic (Sus scrofa domestica) pigs and their hybrids based on polymorphism of a fragment of the D-loop region in the mitochondrial DNA

We examined the variation in mitochondrial DNA by sequencing the D-loop region in wild and domestic (large-white breed) pigs, in hybrids between domestic and wild pigs, and in Monteiro pigs. A D-loop fragment of approximately 330 bp was amplified by PCR. Sequencing of DNA amplicons identified haplotypes previously described as European and Asian types. Monteiro pigs and wild pigs had European haplotypes and domestic pigs had both European and Asian haplotypes. ©FUNPEC-RP.

Uso de reguladores de crescimento como alternativa tecnológica na cultura da macieira ‘Eva’ (Malus domestica Borkh.)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Análise isotópica da variabilidade natural do carbono-13 e análise energética em suco, néctar e refrigerante de maçã (malus domestica, borkh)

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

Diversidade de maçãs (Malus Domestica Borkh.) nas feiras de produtores de Nova Iorque

Visitando feiras de produtores locais na cidade de Nova Iorque por um ano, entre junho de 2008 e maio de 2009, verificou-se significativa amplitude varietal das maçãs, demonstrando-se um aspecto particular da diversidade vegetal. Evidenciadas no verão pela presença volumosa, é no outono e inverno que se nota a diversidade intraespecífica, atravessando todos os períodos sazonais. Verificou-se a existência de mais de quatro centenas de espécies vegetais disponíveis no mercado, escolhendo-se a maçã como um dos tipos mais evidentes para a análise da diversidade, uma vez que se oferecem em 65 cultivares nas feiras do estado ao qual emprestam o apelido. Representam um tipo de diversidade que extrapola ao conceito de especialização, usualmente atribuído à agricultura dos países desenvolvidos, uma vez que convivem com outras espécies nos espaços em que são cultivadas. O objetivo deste artigo é demonstrar a diversidade genética e de usos das maçãs que se expressa na atividade comercial exercida nas ruas e praças nova iorquinas.

Genomic and transcriptional analysis of allergen genes in apple (Malus x domestica)

Apple consumption is highly recomended for a healthy diet and is the most important fruit produced in temperate climate regions. Unfortunately, it is also one of the fruit that most ofthen provoks allergy in atopic patients and the only treatment available up to date for these apple allergic patients is the avoidance. Apple allergy is due to the presence of four major classes of allergens: Mal d 1 (PR-10/Bet v 1-like proteins), Mal d 2 (Thaumatine-like proteins), Mal d 3 (Lipid transfer protein) and Mal d 4 (profilin). In this work new advances in the characterization of apple allergen gene families have been reached using a multidisciplinary approach. First of all, a genomic approach was used for the characterization of the allergen gene families of Mal d 1 (task of Chapter 1), Mal d 2 and Mal d 4 (task of Chapter 5). In particular, in Chapter 1 the study of two large contiguos blocks of DNA sequences containing the Mal d 1 gene cluster on LG16 allowed to acquire many new findings on number and orientation of genes in the cluster, their physical distances, their regulatory sequences and the presence of other genes or pseudogenes in this genomic region. Three new members were discovered co-localizing with the other Mal d 1 genes of LG16 suggesting that the complexity of the genetic base of allergenicity will increase with new advances. Many retrotranspon elements were also retrieved in this cluster. Due to the developement of molecular markers on the two sequences, the anchoring of the physical and the genetic map of the region has been successfully achieved. Moreover, in Chapter 5 the existence of other loci for the Thaumatine-like protein family in apple (Mal d 2.03 on LG4 and Mal d 2.02 on LG17) respect the one reported up to now was demonstred for the first time. Also one new locus for profilins (Mal d 4.04) was mapped on LG2, close to the Mal d 4.02 locus, suggesting a cluster organization for this gene family, as is well reported for Mal d 1 family. Secondly, a methodological approach was used to set up an highly specific tool to discriminate and quantify the expression of each Mal d 1 allergen gene (task of Chapter 2). In aprticular, a set of 20 Mal d 1 gene specific primer pairs for the quantitative Real time PCR technique was validated and optimized. As a first application, this tool was used on leaves and fruit tissues of the cultivar Florina in order to identify the Mal d 1 allergen genes that are expressed in different tissues. The differential expression retrieved in this study revealed a tissue-specificity for some Mal d 1 genes: 10/20 Mal d 1 genes were expressed in fruits and, indeed, probably more involved in the allergic reactions; while 17/20 Mal d 1 genes were expressed in leaves challenged with the fungus Venturia inaequalis and therefore probably interesting in the study of the plant defense mechanism. In Chapter 3 the specific expression levels of the 10 Mal d 1 isoallergen genes, found to be expressed in fruits, were studied for the first time in skin and flesh of apples of different genotypes. A complex gene expression profile was obtained due to the high gene-, tissue- and genotype-variability. Despite this, Mal d 1.06A and Mal d 1.07 expression patterns resulted particularly associated with the degree of allergenicity of the different cultivars. They were not the most expressed Mal d 1 genes in apple but here it was hypotized a relevant importance in the determination of allergenicity for both qualitative and quantitative aspects of the Mal d 1 gene expression levels. In Chapter 4 a clear modulation for all the 17 PR-10 genes tested in young leaves of Florina after challenging with the fungus V. inaequalis have been reported but with a peculiar expression profile for each gene. Interestingly, all the Mal d 1 genes resulted up-regulated except Mal d 1.10 that was down-regulated after the challenging with the fungus. The differences in direction, timing and magnitude of induction seem to confirm the hypothesis of a subfunctionalization inside the gene family despite an high sequencce and structure similarity. Moreover, a modulation of PR-10 genes was showed both in compatible (Gala-V. inaequalis) and incompatible (Florina-V. inaequalis) interactions contribute to validate the hypothesis of an indirect role for at least some of these proteins in the induced defense responses. Finally, a certain modulation of PR-10 transcripts retrieved also in leaves treated with water confirm their abilty to respond also to abiotic stress. To conclude, the genomic approach used here allowed to create a comprehensive inventory of all the genes of allergen families, especially in the case of extended gene families like Mal d 1. This knowledge can be considered a basal prerequisite for many further studies. On the other hand, the specific transcriptional approach make it possible to evaluate the Mal d 1 genes behavior on different samples and conditions and therefore, to speculate on their involvement on apple allergenicity process. Considering the double nature of Mal d 1 proteins, as apple allergens and as PR-10 proteins, the gene expression analysis upon the attack of the fungus created the base for unravel the Mal d 1 biological functions. In particular, the knowledge acquired in this work about the PR-10 genes putatively more involved in the specific Malus-V. inaequalis interaction will be helpful, in the future, to drive the apple breeding for hypo-allergenicity genotype without compromise the mechanism of response of the plants to stress conditions. For the future, the survey of the differences in allergenicity among cultivars has to be be thorough including other genotypes and allergic patients in the tests. After this, the allelic diversity analysis with the high and low allergenic cultivars on all the allergen genes, in particular on the ones with transcription levels correlated to allergencity, will provide the genetic background of the low ones. This step from genes to alleles will allow the develop of molecular markers for them that might be used to effectively addressed the apple breeding for hypo-allergenicity. Another important step forward for the study of apple allergens will be the use of a specific proteomic approach since apple allergy is a multifactor-determined disease and only an interdisciplinary and integrated approach can be effective for its prevention and treatment.

Architectural development and dry matter production in a multisite trial on single and multiaxis apple trees (Malus domestica Borkh) grafted on different rootstocks

In two Italian sites, multiaxis trees slightly reduced primary axis length and secondary axis length of newly grafted trees, and increased the number of secondary shoots. The total length, node production, and total dry matter gain were proportional to the number of axis. Growth of both primary and secondary shoots, and dry matter accumulation, have been found to be also well related to rootstock vigour. A great variability in axillary shoot production was recorded among different environments. Grafted trees had higher primary growth, secondary axis growth, and dry matter gain than chip budded trees. Stem water potential measured in the second year after grafting was not affected by rootstocks or number of leaders. Measurements performed in New Zealand (Hawke’s Bay) during the second year after grafting revealed that both final length and growth rate of primary and secondary axis were related to the rootstock rather than to the training system. Dwarfing rootstocks reduced the number of long vegetative shoots and increased the proportion of less vigorous shoots.