Expressed sequence tags related to nitrogen metabolism in maize inoculated with Azospirillum brasilense.

The relative quantitative real-time expression of two expressed sequence tags (ESTs) codifying for key enzymes in nitrogen metabolism in maize, nitrate reductase (ZmNR), and glutamine synthetase (ZmGln1-3) was performed for genotypes inoculated with Azospirillum brasilense. Two commercial single-cross hybrids (AG7098 and 2B707) and two experimental synthetic varieties (V2 and V4) were raised under controlled greenhouse conditions, in six treatment groups corresponding to different forms of inoculation and different levels of nitrogen application by top-dressing. The genotypes presented distinct responses to inoculation with A. brasilense. Increases in the expression of ZmNR were observed for the hybrids, while V4 only displayed a greater level of expression when the plants received nitrogenous fertilization by top-dressing and there was no inoculation. The expression of the ZmGln1-3EST was induced by A. brasilense in the hybrids and the variety V4. In contrast, the variety V2 did not respond to inoculation.

Canonical correlations between agronomic traits and seed physiological quality in segregating soybean populations.

The objective of this study was to evaluate the relationship between agronomic traits and physiological traits of seeds in segregating soybean populations by canonical correlation analysis. Seven populations and two commercial cultivars in three generations were used: F3 plants and F4 seeds; F4 plants and F5 seeds, and F4 seeds and plants. The following agronomic traits (group I) were evaluated: number of days to maturity, plant height at maturity, insertion height of first pod, number of pods, grain yield, and oil content. The physiological quality of seeds (group II) was evaluated using germination, accelerated aging, emergence, and emergence rate index tests. The results showed that agronomic traits and physiological traits of seeds are not independent. Intergroup associations were established by the first canonical pair for the generation of F3 plants and F4 seeds, especially between more productive plants with a larger pod number and high oil content and seeds with a high germination percentage and emergence rate. For the generation of F4 plants and F5 seeds, the first canonical pair indicated an association between reduced maturity cycle, seeds with a high emergence percentage and a high percentage of normal seedlings after accelerated aging. According to the second canonical pair, more productive and taller plants were associated with seed vigor. For the generation of F4 seeds and plants, the associations established by the first canonical pair occurred between seed vigor and more productive plants with high oil content and reduced maturity cycle, and those established by the second canonical pair between seeds of high physiological quality and tall plants.

Genetic divergence in a soybean (Glycine max) diversity panel based on agro-morphological traits.

Owing to the narrow genetic basis of soybean (Glycine max), the incorporation of new sources of germplasm is indispensable when searching for alleles that contribute to a greater diversity of varieties. The alternative is plant introduction, which may increase genetic variability within breeding programs. Multivariate techniques are important tools to study genetic diversity and allow the precise elucidation of variability in a set of genotypes of interest. The agro-morphological traits of 93 soybean accessions from various continents were analyzed in order to assess the genetic diversity present, and to highlight important traits. The experimental design was incomplete blocks (Alpha lattice, 8 x 12) with three replicates. Nine agro-morphological traits were analyzed, and principal component analysis and cluster analysis were performed, the latter by Ward's method. The dendrogram obtained contained eight subgroups, confirming the genetic diversity among the accessions and revealing similarities between 11 national genotypes. The geographical origin of the accessions was not always related to the clusters. The traits evaluated, and the methods used, facilitated the distinction and characterization of genotypes between and within groups, and could be used in Brazilian soybean breeding programs.

HLA-A, HLA-B and HLA-DRB1 haplotype frequencies in Piauí's volunteer bone marrow donors enrolled at the Brazilian registry.

This study aimed to report the antigen and haplotype frequencies (HFs) of volunteer bone marrow donors (VBMDs) from the state of Piauí who were enrolled in the National Volunteer Bone Marrow Donor Registry (REDOME). The research subjects were 21,943 volunteer bone marrow donors, predominantly young adult women (53.3%). The most frequent allelic group was HLA-A2, followed by -DRB1*13, -DRB1*04, -DRB1*07, -B*15, -B∗35, -B*44, -A*24 and -A*03. Of the 2,704 haplotypes observed, the three most frequent haplotypes were A*29 B*44 DRB1*07 (1.45%), A*01 B*08 DRB1*03 (1.4%) and A*03 B*07 DRB1*15 (0.92%). These three haplotypes were in linkage disequilibrium. PCA showed that 98% of the VBMDs have HLA allele frequencies that are very similar to those from Teresina, the capital city of Piauí. According to the PCA results, these municipalities are distributed with a close proximity to Teresina, which in turn has a close genetic proximity to the Hispanic ethnicity, intermediate proximity to Caucasians and Africans and a distant kinship to Amerindians. The hierarchical proximity of the population of Piauí to the Portuguese and Hispanic populations to shows the strong influence of the latter on the former.

Digenetic trematodes in South American sea lions from southern Brazilian waters.

The aim of this work was to perform a systematic study to detect and quantify the digenetic trematode infections in South American sea lions from the southern Brazilian coast. Twenty-four South American sea lions, Otaria flavescens (Carnivora: Otaridae), were found dead along the coast of Rio Grande do Sul State, Brazil, between June 2010 and September of 2011. Two trematode species were found in the intestines of O. flavescens, i.e., Stephanoprora uruguayense (Digenea: Echinostomatidae) and Ascocotyle (Phagicola) longa (Digenea: Heterophyidae). Ascocotyle (P.) longa reached a prevalence of 33.3% and mean intensity of 248,500, whereas S. uruguayense showed a prevalence of 4.2% and mean intensity of 202. The 2 trematode species infecting sea lions were likely transmitted by feeding on mullets, Mugil platanus, that commonly harbor heterophyid metacercariae. The present work is the first report of digenetic trematodes infecting O. flavescens in Brazil. The high prevalence and mean intensity values of the 2 trematode species infecting sea lions in the present study suggest caution in human consumption of mullets and other fish, which can be infected with the metacercariae of these trematodes known to have zoonotic potential.

Antiallodynic effect and side effects of Phα1ß, a neurotoxin from the spider Phoneutria nigriventer: comparison with ω-conotoxin MVIIA and morphine.

Phα1ß is a potent toxin obtained from the spider Phoneutria nigriventer that blocks neuronal voltage-sensitive Ca(2+) channels. This study compared the antiallodynic effects of Phα1ß, ω-conotoxin MVIIA and morphine in mice and their side effects in rats. Mechanical allodynia was measured in mice receiving single intrathecal administration of Phα1ß, ω-conotoxin MVIIA or morphine before or after the incisional plantar procedure. The effect of the treatments on cardiovascular profile and global neurological were evaluated in rats. The expression of pro or anti-inflammatory cytokines of human polymorph mononuclear cells was also evaluated. Preemptive use of ω-conotoxin MVIIA (1.0 or 10 pmol/site) or morphine (1000 pmol/site) induced shorter antiallodynic effect than Phα1ß (100 pmol/site) in mice. Post-incision administration of Phα1ß (200 pmol/site) induced longer mechanical antiallodynic effect than ω-conotoxin MVIIA (1.0 or 10 pmol/site) or morphine (1000 pmol/site). Intrathecal injection of Phα1ß (200 pmol/site) and morphine (433 pmol/site) did not change while ω-conotoxin MVIIA (100 pmol/site) increased the heart rate in rats 3 h after its administration. Phα1ß (200 pmol/site), ω-conotoxin MVIIA (100 pmol/site) and morphine (433 pmol/site) did not change mean arterial pressure 0.5 and 3 h after their administration. The treatments did not alter neurological performance assessed by global neurological evaluation and open-field test. The tested drugs did not induced expression of pro or anti-inflammatory cytokines in CD4 monocytes. In conclusion, preemptive administration Phα1ß in mice induced longer antiallodynic effect than ω-conotoxin MVIIA and morphine. Phα1ß also induced a longer mechanical antiallodynic effect than ω-conotoxin MVIIA and morphine when used after the surgical incision. The present results suggest that Phα1ß has a potential application in the management of postoperative pain with low side effects.

Orofacial granulomatosis of the lip: a report of 2 cases with histological and immunohistochemical analyses and intralesional corticotherapy.

Orofacial granulomatosis is a generic term applied to manifestations of several diseases including sarcoidosis, Crohn's disease, Melkersson-Rosenthal syndrome, cheilitis granulomatosa of Miescher, tuberculosis and foreign-body reactions. What bonds these diseases together is the presence of noncaseating granulomas. A typical clinical manifestation of orofacial granulomatosis is recurrent labial swellings that eventually persist. This article describes 2 cases of OG diagnosed with the aid of immunohistochemical analysis and successfully treated with intralesional steroids.

In vivo anti-angiogenic effects further support the promise of the antineoplasic activity of methyl jasmonate.

Molecular plant components have long been aimed at the angiogenesis and anti-angiogenesis pathways, and have been tested as sources for antineoplasic drugs with promising success. The present work deals with the anti-angiogenic effects of Methyl Jasmonate. Jasmonate derivatives were demonstrated to selectively damage the mitochondria of cancer cells. In vitro, 1-10 mM Methyl Jasmonate induced the cell death of the human umbilical vein endothelial cells (HUVEC) and the Murine melanoma cells (B16F10), while micromolar concentrations were ineffective. In vivo, comparable concentrations were toxic and reduced the vessel density of the Chorioallantoic Membrane of the Chicken Embryo (CAM). However, 1-10 microM concentrations produced a complex effect. There was increased capillary budding, but the new vessels were leakier and less organised than corresponding controls. It is suggested that not only direct toxicity, but also the drug effects upon angiogenesis are relevant to the antineoplasic effects of Methyl Jasmonate.

PHA(MCL) biosynthesis systems in Pseudomonas aeruginosa and Pseudomonas putida strains show differences on monomer specificities.

The production of PHA from plant oils by Pseudomonas species soil isolated from a sugarcane crop was evaluated. Out of 22 bacterial strains three were able to use efficiently plant oils to grow and to accumulate PHA. Pseudomonas putida and Pseudomonas aeruginosa strains produced PHA presenting differences on monomer composition compatible with variability on monomer specificity of their PHA biosynthesis system. The molar fraction of 3-hydroxydodecanoate detected in the PHA was linearly correlated to the oleic acid supplied. A non-linear relationship between the molar fractions of 3-hydroxy-6-dodecenoate (3HDdDelta(6)) detected in PHA and the linoleic acid supplied was observed, compatible with saturation in the biosynthesis system capability to channel intermediate of beta-oxidation to PHA synthesis. Although P. putida showed a higher 3HDdDelta(6) yield from linoleic acid when compared to P. aeruginosa, in both species it was less than 10% of the maximum theoretical value. These results contribute to the knowledge about the biosynthesis of PHA with a controlled composition from plant oils allowing in the future establishing the production of these polyesters as tailor-made polymers.

Metaplastic breast carcinomas are basal-like tumours.

AIMS: Recently, an immunohistochemical panel comprising antibodies against HER2, oestrogen receptor (ER), epidermal growth factor receptor (EGFR) and cytokeratin (CK) 5/6 was reported to identify basal-like breast carcinomas, as defined by cDNA microarrays. Our aim was to analyse a series of metaplastic breast carcinomas (MBCs) using this panel plus two other basal markers (CK14 and p63) and progesterone receptor (PR), to define how frequently MBCs show a basal-like immunophenotype. METHODS AND RESULTS: Sixty-five cases were retrieved from the pathology archives of the authors' institutions and reviewed by three of the authors. Immunohistochemistry with antibodies for HER2, ER, EGFR, CK5/6, CK14 and p63 was performed according to standard methods. All but six cases (91%) showed the typical immunoprofile of basal-like tumours (ER- and HER2-, EGFR+ and/or CK5/6+). When CK14 and p63 were added to the panel, two additional cases could be classified as basal-like. The majority of MBCs lacked PR, except 4/19 (21%) carcinomas with squamous metaplasia. CONCLUSIONS: Our results demonstrate that MBCs show a basal-like phenotype, regardless of the type of metaplastic elements. Moreover, as these neoplasms frequently overexpress EGFR (57%), patients with MBC may benefit from treatment with anti-EGFR drugs.

EGFR amplification and lack of activating mutations in metaplastic breast carcinomas.

Metaplastic breast carcinomas are reported to harbour epidermal growth factor receptor (EGFR) overexpression in up to 80% of the cases, but EGFR gene amplification is the underlying genetic mechanism in around one-third of these. In this study, EGFR gene amplification as defined by chromogenic in situ hybridization and protein overexpression was examined in a cohort of 47 metaplastic breast carcinomas. Furthermore, the presence of activating EGFR mutations in exons 18, 19, 20, and 21 was investigated. Thirty-two cases showed EGFR overexpression and of these, 11 (34%) harboured EGFR gene amplification. In addition, EGFR amplification showed a statistically significant association with EGFR overexpression (p < 0.0094) and was restricted to carcinomas with homologous metaplasia. Ten cases, five with and five without EGFR amplification, were subjected to microarray-based CGH, which demonstrated that EGFR copy number gain may occur by amplification of a discrete genomic region or by gains of the short arm of chromosome 7 with a breakpoint near the EGFR gene locus, the minimal region of amplification mapping to EGFR, LANCL2, and SEC61G. No activating EGFR mutations were identified, suggesting that this is unlikely to be a common alternative underlying genetic mechanism for EGFR expression in metaplastic breast carcinomas. Given that metaplastic breast carcinomas are resistant to conventional chemotherapy or hormone therapy regimens and that tumours with EGFR amplification are reported to be sensitive to EGFR tyrosine kinase inhibitors, these findings indicate that further studies are warranted to explore EGFR tyrosine kinase inhibitors as potential therapeutic agents for metaplastic breast carcinomas harbouring amplification of 7p11.2.

Polymorphous low-grade adenocarcinoma of the salivary glands with transformation to high-grade carcinoma.

AIMS: Polymorphous low-grade adenocarcinoma of the minor salivary glands is an infiltrative neoplasm characterized by bland-looking tumour cells arranged in diverse architectural patterns. It is considered to be of low-grade malignant potential in that nodal metastases are seen in only a minority, and distant spread is rare. Even more unusual is the transformation of polymorphous low-grade adenocarcinoma to a histologically high-grade carcinoma, i.e. dedifferentiation. In this paper, we describe the clinicopathological and immunohistochemical findings in two further examples. METHODS AND RESULTS: Two patients presented each with a tumour of the palate. Histopathological examination showed the typical morphological, cytological and immunohistochemical features of a polymorphous low-grade adenocarcinoma. In one case there was a second component of high-grade carcinoma showing nuclear atypia, markedly increased mitotic activity and MIB1 index, as well as prominent zones of necrosis. It expressed epithelial markers and androgen receptors, and thus resembled salivary duct carcinoma. Similar tumour tissue was observed in one of the cervical nodal metastases, which was biopsied at the same time as the palate. In the second patient, a high-grade component was discovered when the tumour recurred in the palate 13 years after the initial biopsy. Whilst morphologically similar to that in first case, there were significant immunohistochemical differences such as retention of some of the polymorphous low-grade adenocarcinoma profile and absence of androgen receptor expression. CONCLUSIONS: Polymorphous low-grade adenocarcinoma was first described relatively recently, and as experience with it continues to accumulate, it is becoming clear that late recurrences and metastases, whilst still infrequent, may not be quite as rare as previously thought. Reports of histological transformation are even scarcer, and most occurred at least 13 years after the polymorphous low-grade adenocarcinoma was initially recognized. It is a real possibility that this phenomenon, like clinical progression, may also be encountered more often as time passes. Therefore, we believe that, whilst polymorphous low-grade adenocarcinoma is certainly far less aggressive than, for example, adenoid cystic carcinoma, it nevertheless remains a true malignancy with a potential to prove fatal in a minority of patients.

The complete nucleotide sequences of L3 and S7 segments of Ibaraki virus encoding for the major inner capsid proteins, VP3 and VP7.

The complete nucleotide sequences of the genes encoding two of the major inner capsid proteins of Ibaraki virus (IBAV), belonging to epizootic hemorrhagic disease virus serotype 2 (EHDV-2) were determined. The L3 RNA segment is 2768 nucleotides in length which encodes VP3 polypeptides of 899 amino acid residues (M.W. 103 kDa). The S7 RNA segment, which encodes the VP7 core protein, is 1162 nucleotides in length and encodes 349 amino acids (M.W. 38 kDa). These RNA segments had the characteristic consensus motifs of Orbivirus RNA segments in termini, namely 5'-GUUAAA... and ...ACUUAC-3'. The comparison of the IBAV L3 and S7 sequences with those of other two EHDV-2 isolates revealed the higher homologies of 93% and 92% against EHDV-2 Australia isolate (EHDV-2AUS) and lower homologies of 80% and 81% against EHDV-2 North America isolate, respectively. The phylogenetic analysis based on L3 and S7 genes also indicated close relationships between IBAV and EHDV-2AUS.

Atividade antimiocrobiana de bactérias lácticas isoladas de produtos cárneos frente a Listeria monocytogenes e Staphylococcus aureus / Antimicrobial activity of lactic acid bacteria isolated from meat products against Listeria monocytogenes and Staphylococcus aureus

De seis amostras de pernil desossado curado e três amostras de salame tipo italiano artesanal foram isoladas 288 cepas de bactérias lácticas. Pelo teste de inibiçäo direta 59 delas foram capazes de inibir in vitro o desenvolvimento de cepas de Listeria monocytogenes e de Staphylococcus aureus, o que demonstra que produzem compostos inibitórios

CD99/MIC-2 surface protein expression in breast carcinomas.

AIM: To evaluate the expression of CD99/MIC-2 surface protein in invasive breast carcinomas and demonstrate whether or not there is a relationship with tumour phenotype. METHODS AND RESULTS: Thirty-five invasive breast carcinomas, including five metaplastic carcinomas, were stained with CD99 primary antibodies using standard protocols based on streptavidin-biotin-peroxidase method. Four out of five metaplastic carcinomas expressed CD99/MIC-2 protein, three of them were matrix-producing carcinomas. From the other 30 cases, only an invasive apocrine carcinoma was positive. There was no statistical correlation between CD99 expression and the parameters analysed (histological typing and grading, proliferative index and nodal status). CONCLUSIONS: CD99/MIC-2 is expressed in breast carcinomas, especially in the matrix-producing variant of metaplastic carcinomas, which impairs its use as a marker to differentiate metaplastic carcinomas from primary and metastatic sarcomas of the breast. It seems to have no prognostic implications. However, phenotype similarities with other chondromyxoid tumours that also express the protein, like mesenchymal chondrosarcomas, suggest a relationship between MIC-2 reactivity and morphological differentiation.

[Experiencing diversity through home care of the elderly--challenges and advances]. / Crescendo na diversidade pelo cuidado domiciliar aos idosos--desafios e avanços.

The present article is a study about the nursing care of elderly people. The diversity of experiences generated by the care of the elderly in their homes has opened news area of research, improved and promoted the nursing care. The objective of this work is to characterize the home-care of elderly as an area of interest for the work of the nursing professionals. The study describes aspects of the dynamics of this work based on a critical analysis of an experience carried out with a group of elderly and pointing out its advances and challenges. The methodology used in this investigation were records of the experience of some elderly people who had undergone surgery and were still in need of nursing care at their homes. Results show that there were improvements in terms of the nursing practices, especially the practices of educational and ethic-legal nature. The practices of educational nature could propitiate a sense of citizenship and freedom, which are prerogatives for the lives of these people. The ethic-legal practices promoted more involvement of the nursing professionals in the interdisciplinary team.

Mimics of copper proteins: structural and functional aspects.

The importance of copper as an essential element can be estimated by the wide range of copper proteins and enzymes playing different roles in biological systems. In the last decades many bioinorganic studies were developed on mimetic complexes of copper-dependent proteins, in order to verify the interrelations between structural and functional properties of active copper centers. Among the most studied copper ion ligand, diimine compounds have deserved special attention due their flexibility, facility of preparation, and ability to stabilize both oxidation states of this metal. In our laboratory, we have been investigating some Schiff base copper complexes as mimics of different proteins, with emphasis on functional aspects, trying to elucidate mechanisms of reaction, based on proposed intermediary species, in addition to molecular shapes. Particularly, mimics of the copper-zinc superoxide dismutase, and of monooxigenases and oxidases exhibiting dicopper sites are discussed in this work.

Cloning and expression of the M5 RNA segment encoding outer capsid VP5 of epizootic hemorrhagic disease virus Japan serotype 2, Ibaraki virus.

The complete nucleotide sequence of a cDNA clone representing the M5 RNA segment of epizootic hemorrhagic disease virus Japan serotype 2 (EHDV-2), Ibaraki virus, was determined. The M5 segment is 1641 base pairs long with the single open reading frame which predicts a polypeptide of 527 amino acids. The comparison of the amino acid sequence of the VP5 with those of EHDV-1, bluetongue virus serotype 10, and African horse sickness virus serotype 4 revealed that the protein shared 67%, 57% and 42% homologies, respectively. In addition, the VP5 protein was expressed in insect cells by recombinant baculovirus, which could be recognized by the mouse anti-EHDV-2 sera at a position of the expected 59 kDa on immunoblot analysis.

The complete nucleotide sequence of segment L2 of Ibaraki virus encoding for the antigen recognized by neutralizing antibodies.

The complete nucleotide sequence of cDNA clones representing the L2 dsRNA from Japan isolate of epizootic hemorrhagic disease serotype 2 (EHDV-2JPN) was determined. The EHDV-2JPN L2 gene is 3002 base pairs long with a single open reading frame of 2949 bp which predicts a polypeptide of 982 amino acid residues. Comparison of VP2 sequence between Japan and North American Isolates of EHDV-2 showed a 72% homology in spite of the same serotype, although those among the North American isolates showed a high genetic identity (>97%).

Stimulation of immunoreactive insulin release by glucose in rat brain synaptosomes.

The effect of glucose on the release of immunoreactive insulin (IRI) in synaptosomes isolated from rat brain was studied. In the absence of glucose synaptosomes release about 4% (0.77 microIU/mg protein) of total content. Glucose increases significantly the IRI released by synaptosomes. Addition of the glycolytic inhibitor iodoacetic acid (IAA), decreased the glucose-induced release of IRI by about 50%, suggesting that glucose metabolism is involved. The observation that glucose provides a concentration related signal for IRI release indicates that this synaptosomal preparation may be useful as a model for research on the mechanism of insulin release in brain.