Histopathology of gills of Piaractus mesopotamicus (Holmberg, 1887) and Prochilodus lineatus (Valenciennes, 1836) infested by monogenean and myxosporea, caugth in Aquidauana River, State of Mato Grosso do Sul, Brazil

Realizou-se análise histológica de brânquias de 15 espécimes de Piaractus mesopotamicus e 19 Prochilodus lineatus coletados de abril a novembro de 2004, no Rio Aquidauana, MS, com intuito de contribuir com achados anatomopatológicos em brânquias dessas espécies de peixes de água doce. Amostras de brânquias foram fixadas em formalina 10%, tamponadas e processadas conforme rotina histológica. em P. mesopotamicus observou-se presença de monogênea e cistos de mixosporídio da espécie Henneguya piaractus, com localização intralamelar em vários estágios de desenvolvimento, localizados em todas as regiões (basal, mediana ou distal) das lamelas. Cistos intraepiteliais causaram dilatação e deformação das lamelas vizinhas. em brânquias de P. lineatus, observou-se presença de monogênea. Nas duas espécies de hospedeiro foram registradas hiperplasia do epitélio branquial e desorganização estrutural das lamelas em extensas regiões, alterações que causaram a fusão lamelar. em poucos casos registrou-se presença de células inflamatórias mononucleares e focos hemorrágicos na região distal das lamelas.

Experimental Salmonella Gallinarum infection in light laying hen lines

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

A novel A2 allele found in Leishmania (Leishmania) infantum chagasi

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

A defective mutant of Salmonella enterica Serovar Gallinarum in cobalamin biosynthesis is avirulent in chickens

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

Colonização e lesão em fêmeas ingurgitadas do carrapato Rhipicephalus sanguineus causadas pelo fungo Metarhizium anisopliae

O presente trabalho teve como objetivo verificar a forma de penetração do fungo Metarhizium anisopliae [METSCH. (SOROKIN, 1883)] em carrapatos da espécie Rhipicephalus sanguineus (LATREILLE, 1806), assim como as lesões infringidas nos tecidos internos do ácaro. A forma de aderência e penetração do fungo foi estudada através da microscopia eletrônica de varredura e a ação do fungo nos tecidos internos avaliada em secções histológicas convencionais. Para observação destes eventos, realizaram-se infecções experimentais em 11 grupos de fêmeas ingurgitadas do carrapato R. sanguineus contendo 12 fêmeas ingurgitadas cada. Para tal, as fêmeas ingurgitadas foram banhadas durante 3 minutos, sob agitação manual, em suspensão com concentração 108 conídios/mL. No caso dos grupos controle o banho foi realizado apenas no veículo da suspensão. Os carrapatos foram processados para histopatologia e microscopia eletrônica em diversos tempos após a infecção, a saber: 1 e 18h, e um, dois, três, quatro, cinco, seis, sete, nove e onze dias. Observou-se que a maior parte dos conídios germinou em até 18h após a inoculação e que o fungo penetrou no ácaro através do tegumento 48h após a infecção. Após a penetração, o fungo invadiu o corpo do hospedeiro promovendo uma colonização difusa, sem preferência aparente por tecidos específicos. Dentre as lesões nos tecidos internos do ácaro, ressalta-se o rompimento da parede intestinal e vazamento do conteúdo para a hemocele. A morte do hospedeiro ocorreu entre 96 e 120h pós-infecção, e a esporulação do patógeno sobre o cadáver do ácaro iniciou-se em torno de 120 a 144h pós-infecção. Espera-se, com este trabalho, contribuir para o desenvolvimento e viabilização de técnicas de controle biológico dos carrapatos por fungos como alternativa ao uso de acaricidas.

Um estudo de metamaterial em antenas de microfita

Metamaterials have attracted a great attention in recent years mostly due to their electromagnetic properties not found in nature. Since metamaterials began to be synthesized by the insertion of artificially manufactured inclusions in a medium specified host , it provides the researcher a broad collection of independent parameters such as the electromagnetic properties of the material host. In this work was presents an investigation of the unique properties of Split Ring Resonators and compounds metamaterials was performed. We presents a theoretical and numerical analysis , using the full-wave formalism by applying the Transverse Transmission Line - LTT method for the radiation characteristics of a rectangular microstrip antenna using metamaterial substrate, as is successfully demonstrated the practical use of these structures in antennas. We experimentally confirmed that composite metamaterial can improved the performance of the structures considered in this thesis

Estudo Comparativo da Biologia de Meloidogyne enterolobii (= M. mayaguensis) e Meloidogyne javanica em tomateiros com gene Mi

Visando conhecer a relação parasito hospedeiro de M. enterolobii em plantas com resistência a nematoides das galhas, estudos comparativos da biologia de M. enterolobii e M. javanica em tomateiros com o gene Mi foram conduzidos. O experimento foi conduzido em esquema fatorial 2x2, composto de dois porta-enxertos de tomateiro ('Magnet' e 'Helper M') e duas espécies de nematoides das galhas (M. enterolobii e M. javanica) com cinco repetições. As plantas foram inoculadas com 500 juvenis infectantes (J2) de M. enterolobii ou M. javanica. As raízes foram coletadas aos 3, 10, 17, 24 e 31 dias após a inoculação, coloridas com fucsina ácida, e dissecadas sob microscópio estereoscópico para a localização e contagem dos diferentes estádios de desenvolvimento dos nematoides. Os dados foram submetidos à análise de variância e as médias comparadas pelo teste de Tukey a 5% de probabilidade. Os resultados obtidos mostraram que, embora ambas as espécies tenham sido capazes de penetrar as raízes dos porta-enxertos de tomateiro, somente M. enterolobii conseguiu desenvolver-se normalmente, com as fêmeas maduras realizando suas posturas, a partir de 24 dias após a inoculação.

Avaliação de dois clones de Escherichia coli recombinante quanto ao crescimento e expressão de antígenos de Leishmania chagasi (kmp11 e P36)

Visceral leishmaniosis caused by Leishmania chagasi, also known as calazar, presented, in the period from 1990 to 2005, tax of incidence in Brazil varying between 1 and 3 cases for 100 000 inhabitants. The Northeast region that up to the year of 2000 contributed with almost 90% of the registered cases is reducing his participation in the current decade, reaching 56% in 2005. Conventional leishmaniasis treatment is costly and it shows high toxicity, demanding more research for alternative treatments, with special interest in development of vaccines and diagnosis kits which include production of recombinant antigens by host cells. Escherichia coli has been the microorganism most studied and used as a host for recombinant protein production. Therefore, the aim of this work was to study the influence of induction on cellular growth and to verify the type of Leishmania chagasi antigens expression (intra or extracellular) during two recombinant E. coli clones (kmp11 and P36) cultivation in rotary incubator (shaker) using three different media (2xTY, TB, FASS+EL). For that, tests were carried out using conditions established in the literature for E. coli (37°C and 200 rpm) and media supplemented with antibiotics to guarantee that only competent cells grows. First, tests were carried out without induction in order to verify the two microorganisms kinetic behavior (growth and substrate consumption) in different media. Next, the induction was carried out through the addition of IPTG (1mM as final concentration), at the first hour of cultivation. It was observed that protein expression were intracellular for all clones and media tested, however the highest level of expression was clearly observed by the electrophoresis band density (intensity) for 2xTY medium and kmp11 protein. Although it contains the lowest substrate concentration, consequently, a reduced cellular concentration when compared to other media, it appeared that this medium and clone combination is the most indicated for recombinant protein production. Therefore, the objective of this work was achieved, since the interested proteins were produced. Consequently, this result motivates new studies for production optimization using different cultivation strategies

Behavior of Leishmania major metacyclic promastigotes during the course of infection and immune response development in resistant versus susceptible hosts

Pouco se conhece sobre os epítopos derivados de promastigotas metacíclicos de Leishmania que são importantes para a regulação ou destruição do parasita, como alvos de ação imunológica no hospedeiro vertebrado. Neste estudo, nós investigamos um método alternativo para obter promastigotas metacíclicos de Leishmania major, pela avaliação do curso da infecção e reação de hipersensibilidade do tipo retardado (HTR) em hospedeiros resistentes e susceptíveis. Promastigotas não-infectantes (procíclicos) de L. major, recentemente isolados de amastigotas, foram selecionados pela adesão a colunas de lã de vidro negativamente carregadas, enquanto que promastigotas metacíclicos não se aderem à coluna e podem ser recuperados com facilidade. Condições ótimas de cromatografia foram validadas por análise estatística. O rendimento médio de parasitas obtidos após separação em colunas de lã de vidro e a viabilidade dos promastigotas foram estimados por microscopia óptica. Os promastigotas metacíclicos tiveram um rendimento médio de 43,5% a 57,5%. Camundongos BALB/c (susceptíveis) e camundongos C57BL/6 (resistentes) apresentaram padrões distintos de lesões cutâneas, os primeiros com lesões mais agressivas, induzidas por promastigotas metacíclicos. As respostas à reação de HTR foram maiores nos grupos de camundongos C57BL/6, submetidos à infecção com promastigotas metacíclicos. Estes resultados indicam que o novo método poderia ser integrado aos protocolos existentes para estudar a metaciclogênese de parasitas do gênero Leishmania in vivo.

Associação entre polimorfismos em enzimas de reparo de DNA e ocorrência de meningite bacteriana.

Despite advances in vaccine development and therapy, bacterial meningitis (BM) remains a major cause of death and long-term neurological disabilities. As part of the host inflammatory response to the invading pathogen, factors such as reactive oxygen species are generated, which may damage DNA and trigger the overactivation of DNA repair mechanisms. It is conceivable that the individual susceptibility and outcome of BM may be in part determined by non synonymous polymorphisms that may alter the function of crucial BER DNA repair enzymes as PARP-1, OGG-1 and APE-1. These enzymes, in addition to their important DNA repair function, also perform role of inflammatory regulators. In this work was investigated the non synonymous SNPs APE-1 Asn148Glu, OGG-1 Ser326Cys,PARP-1 Val762Ala, PARP-1 Pro882Leu and PARP-1 Cys908Tyr in patients with bacterial meningitis (BM), chronic meningitis (CM), aseptic meningitis (AM) and not infected (controls). As results we found increased frequency of variant alleles of PARP-1 Val762Ala (P = 0.005) and APE-1 Asn148Glu (P=0.018) in BM patients, APE-1 Asn148Glu in AM patients (P = 0.012) and decrease in the frequency of the variant allele OGG-1 Ser326Cys in patients with CM (P = 0.013), regarding the allelic frequencies in the controls. A major incidence of individuals heterozygous and/ or polymorphic homozygous in BM for PARP-1 Val762Ala (P= 0.0399, OD 4.2, 95% IC 1.213 -14.545) and PARP-1 Val762Ala/ APE-1 Asn148Glu (P = 0.0238, OD 11.111, 95% IC 1.274 - 96.914) was observed related to what was expected in a not infected population. It was also observed a major incidence of combined SNPs in the BM patients compared with the control group (P=0.0281), giving evidences that SNPs can cause some susceptibility to the disease. This combined effect of SNPs seems to regulate the principal cytokines and other factors related to BM inflammatory response and point the importance of DNA repair not only to repair activity when DNA is damaged, but to others essential functions to human organism balance.

Prevalência e susceptibilidade antimicrobiana de Staphylococcusspp. em quadros de saúde e doença periodontal

The aim of this study was determine the prevalence and antimicrobial susceptibility of Staphylococcus spp. from patients with periodontal disease and periodontally healthy, correlate them with factors to host, local environment and traits of the diseases. To this, thirty adults from 19 to 55 years old were selected. They had not periodontal treatment and no antibiotic or antimicrobial was administered during three previous months. From these individuals, sites periodontally healthy, with chronic gingivitis and/or periodontitis were analyzed. Eighteen subgingival dental biofilm samples were collected through sterile paper points being six from each tooth randomly selected, representing conditions mentioned. They were transported to Oral Microbiology laboratory, plated onto Mannitol Salt Agar (MSA) and incubated at 370C in air for 48 h. Staphylococcus spp. were identified by colonial morphology, Gram stain, catalase reaction, susceptibility to bacitracin and coagulase activity. After identification, strains were submitted to the antibiotic susceptibility test with 12 antimicrobials, based on Kirby-Bauer technique. To establish the relation between coagulase-negative Staphylococcus (CSN) presence and their infection levels and host factors, local environment and traits of diseases were used Chi-square, Mann-Whitney and Kruskal-Wallis tests to a confidence level of 95%. 86,7% subjects harbored CSN in 11,7% periodontal sites. These prevalence were 12,1% in healthy sites, 11,7% in chronic gingivitis, 13,5% in slight chronic periodontitis, 6,75% in moderate chronic periodontitis and in sites with advance chronic periodontitis was not isolated CSN, without difference among them (p = 0,672). There was no significant difference to presence and infection levels of CSN as related to host factors, local environm ent and traits of the diseases. Amongst the 74 samples of CSN isolated, the biggest resistance was observed to penicillin (55,4%), erythromycin (32,4%), tetracycline (12,16%) and clindamycin (9,4%). 5,3% of the isolates were resistant to oxacilin and methicillin. No resistance was observed to ciprofloxacin, rifampicin and vancomycin. It was concluded that staphylococci are found in low numbers in healthy or sick periodontal sites in a similar ratio. However, a trend was observed to a reduction in staphylococci occurrence toward more advanced stages of the disease. This low prevalence was not related to any variables analyzed. Susceptibility profile to antibiotics demonstrates a raised resistance to penicillin and a low one to methicillin. To erythromycin, tetracycline and clindamycin was observed a significant resistance

Avaliação de alterações orais em pacientes submetidos a transplante de medula óssea

Bone marrow transplantation (BMT) is currently the best therapeutic option for patients with hematologic diseases, solid tumors or autoimmune disorders. It is characterized by intravenous infusion of hematopoietic stem cells in order to restore marrow function. However, this procedure requires concomitant immunosuppression treatment, which favors the development of certain complications, often manifested in the oral cavity. This study aimed to evaluate the incidence of oral changes in patients undergoing BMT and to correlate these results with clinical aspects related to the patients and the transplants performed. This is a prevalence study, with cross-sectional design, carried out in a BMT service at the Institute of Onco-Hematology of Natal (ION) and Natal Hospital Center. Data collection was based on questionnaires, clinical examination of the oral cavity and consultation in the medical records. The sample consisted of 51 patients undergoing BMT. After the analysis, was found a general status with good health conditions and presence of oral changes in about half of patients who composed the sample. The manifestations observed were, in decreasing order of frequency: mucositis; gingival alteration and thrombocytopenic purpura; mucosal pigmentation; lichenoid reaction and candidiasis. The oral changes were observed more frequently in cases of allogeneic TMO, in different periods post-transplant, without significant differences related to the source of cells. It was found statistically significant association between the presence of graft-versus-host disease (GVHD) and oral changes (p < 0,001). Therefore, it is concluded that there is a relatively high incidence of changes in oral cavity of patients receiving bone marrow transplantation, a fact which confirms the need to consider this site for examination, diagnosis, treatment and prognosis of possible complications of BMT

Análise imuno-histoquímica das proteínas VEGF e HIF-1α na doença periodontal

Periodontal disease is a complex inflammatory and infectious condition that immune host, front of the microbial aggressions, can lead to disease progression, resulting in tissue destruction. The tissue damage induces the release of regulatory molecules, which protective roles and / or destructive, including proteins VEGF (vascular endothelial growth factor vascular) and HIF-1 α (hypoxia-induced factor α -1). Thus, this study investigated, quantitatively and comparatively, the immunohistochemical expression of VEGF (vascular endothelial growth factor) and HIF-1 α (hypoxia induced factor 1-α), proteins involved in inflammation, angiogenesis and hypoxia, in human gingival tissues. Therefore, 75 samples of gingival tissues were examined. Thirty samples were chronic periodontitis, 30 with chronic gingivitis and 15 healthy gingival. After sections analysis, positives and negatives inflammatory and endothelial cells in the connective tissue were counted and converted into percentage. Data were statistically analyzed using Kruskal-Wallis test and Spearman correlation. The results showed that both proteins marked. It was observed higher immunoreactivity for HIF-1 α at chronic gingivitis and periodontitis specimens compared to healthy sites, however, no statistically significant differences were observed among them (p>0.05). The VEGF immunostaining showed similarity among the cases of periodontitis, gingivitis and healthy gingiva. Moderate and positive correlation statistically significant differences were verified for the expressions of VEGF and HIF-1α in gingival health (r = 0,529, p = 0.04). Thus, it can be conclude that possibly the route of HIF-1 α, is activated in periodontal disease may have involvement of the protein in pathogenesis and progression of disease, and that activation of VEGF, can be in addition to being triggered transcription by HIF-1 α may be also influenced by other additional factors such as the action of periodontal microorganisms endotoxins

Resposta celular Th1 /Th2 na doença periodontal experimental, em ratos: um estudo imuno-histoquímico

Host response plays a major role in the pathogenesis of periodontal disease. Mediators such as inflammatory cytokines which are secreted during the immune response to bacterial challenges have ambiguous functions that may or may not lead to protection of the attacked tissue. In this context, experimental evidence suggests that T-helper 1 (Th-1) and T-helper 2 (Th-2) mediated responses are potentially important during the disease process. The aims of this study therefore were to further clarify the role played by Th2 cells during different time points of the active phase of periodontal disease, as well as, to investigate whether there was any evidence of a Th1 response in the periodontal disease microenvironment. Experimental periodontitis was induced in 30 Wistar male rats by placing cotton ligatures around the mandibular first molars. The rats were then randomly divided into two groups. Group1 (G1=15) and Group 2 (G2=15). In G1 the ligatures were maintained for 2 days, whereas in G2 the ligatures were left for 15 days, a time point that corresponds to the advanced stage of periodontal disease The contra-lateral teeth served as controls (no ligatures). Immunohistochemical investigation for the presence in gingival tissue of Th2 specific transcription factor (GATA3) and the subunit of the IFN-γ receptor was carried out after the disease induction period. Light microscopy analysis revealed a decrease in the expression of GATA-3 as bone loss progressed. On the other hand, although IFN-γ R1 was detected at an early stage of the active phase of disease its expression remained unaltered during the remaining period of the study. These results indicate that the Th2 response have a protective role during the pathogenesis of periodontal disease and that the progression of the periodontal disease is related with the unbalance of the responses Th1/Th2

Análise imuno-histoquímica das metaloproteinases da matriz ( MMP-1,MMP-2 e MMP-9) na doença periodontal

The tissular destruction found in periodontal diseases is caused mainly by components of the host that have its production stimulated by the products of the microorganisms present on the plaque. Matrix Metalloproteinases (MMPs), a class of enzymes involved both in physiologic and pathologic extracellular matrix degradation are considered the main responsible for the characteristic tissular loss in periodontal disease, and the understanding of how this happens can have a series of beneficial implications for prevention, diagnosis and treatment of this illness. The aim of this work was to study the immunohistochemical expression of MMP-1, MMP-2, and MMP-9 in fragments of gingival biopsies with clinical diagnose of periodontal disease. MMP-1 has expressed significantly more than the others MMPs in gingivitis both in the epithelium (p=0,0008) and connective tissue (p=0,0049). In periodontitis, both MMP-1 and MMP-9 has expressed significantly more than MMP-2 in the epithelium (p<0,0001) and in the connective tissue (p=0,0002). MMP-1 and MMP-9 presented more expression in periodontitis than in gingivitis but MMP-1 only in the connective tissue (p=0,03) and MMP-9 in the epithelium (p=0,003) and in the connective tissue (p=0,04). In conclusion, these results indicate that the MMP-1 presents high expression in every stages of the periodontal diseases, and increases its expression in the connective tissue when the gingivitis evolves to periodontitis. Therefore, it may have an important role in connective tissue degradation and bone loss observed in disease, since early, in gingivitis, until late stages, in periodontitis, of the periodontal disease. MMP-9 has expressed more in periodontitis than in gingivitis, both in epithelium and in connective tissue. It means that this enzyme may have some importance in the progression of gingivitis to periodontitis by acting in bone resorption observed in this desease