Sequence analysis of the internal transcribed spacer 2 (ITS2) region of rDNA for identifying Trichogramma species and evaluating genetic diversity / Análise de sequência do rDNA da região espaçadora do transcrito interno 2 (ITS2) para identificar espécies de Trichogramma e avaliar a diversidade genética

Abstract Species of Trichogramma Westwood, 1833 (Hymenoptera: Trichogrammtidae) are frequently used as biological control agents against Lepidoptera, but practical application of these egg endoparasitoids are complicated because of their complex taxonomy. This study aimed to compare sequences of internal transcribed spacer regions of ribosomal DNA (ITS2-rDNA) of Trichogramma accessions with those deposited in GenBank in order to access the reliability of the ITS2 as a barcode for discriminating species and evaluating the genetic diversity. ITS2-rDNA sequences obtained from seventeen specimens of Trichogramma confirmed previous identifications based on morphological characteristics. Multiple sequence alignment revealed the existence of highly conserved regions in ITS2 sequences while the neighbour-joining dendrogram indicated that the specimens formed three clusters comprising T. manicobai and T. marandobai (group I), T. galloi (group II) and T. pretiosum (group III). The ITS2 marker was shown to be a powerful DNA barcode for discriminating Trichogramma species and could be used to complement the morphological approach.

Resumo Espécies de Trichogramma Westwood, 1833 (Hymenoptera: Trichogrammatidae) são freqüentemente usadas como agentes de controle biológico contra Lepidoptera, esses endoparasitóides de ovos apresentam taxonomia complexa, o que dificulta sua aplicação prática. Este estudo teve como objetivo comparar seqüências de regiões espaçadoras internas transcritas de DNA ribossômico (ITS2-rDNA) de acessos de Trichogramma com aquelas depositadas no GenBank, a fim de avaliar a confiabilidade do ITS2 barcode para discriminar espécies e avaliar a diversidade genética. As seqüências de ITS2-rDNA obtidas de dezessete espécimes de Trichogramma confirmaram identidades anteriores com base em características morfológicas. O alinhamento de múltiplas sequências revelou a existência de regiões altamente conservadas nas sequências ITS2, enquanto o dendrograma indicou que os espécimes formavam três grupos compreendendo T. manicobai e T. marandobai (grupo I), T. galloi (grupo II) e T. pretiosum (grupo III). O marcador ITS2 mostrou ser um poderoso DNA barcode para discriminar espécies de Trichogramma podendo ser usado como complemento da abordagem morfológica.

Impact of chronic obstructive pulmonary disease on linear and nonlinear dynamics of heart rate variability in patients with heart failure

The objective of this study was to investigate the impact of chronic obstructive pulmonary disease (COPD)-heart failure (HF) coexistence on linear and nonlinear dynamics of heart rate variability (HRV). Forty-one patients (14 with COPD-HF and 27 HF) were enrolled and underwent pulmonary function and echocardiography evaluation to confirm the clinical diagnosis. Heart rate (HR) and R-R intervals (iRR) were collected during active postural maneuver (APM) [supine (10 min) to orthostasis (10 min)], respiratory sinus arrhythmia maneuver (RSA-M) (4 min), and analysis of frequency domain, time domain, and nonlinear HRV. We found expected autonomic response during orthostatic changes with reduction of mean iRR, root mean square of successive differences between heart beats (RMSSD), RR tri index, and high-frequency [HF (nu)] and an increased mean HR, low-frequency [LF (nu)], and LF/HF (nu) compared with supine only in HF patients (P<0.05). Patients with COPD-HF coexistence did not respond to postural change. In addition, in the orthostatic position, higher HF nu and lower LF nu and LF/HF (nu) were observed in COPD-HF compared with HF patients. HF patients showed an opposite response during RSA-M, with increased sympathetic modulation (LF nu) and reduced parasympathetic modulation (HF nu) (P<0.05) compared with COPD-HF patients. COPD-HF directly influenced cardiac autonomic modulation during active postural change and controlled breathing, demonstrating an autonomic imbalance during sympathetic and parasympathetic maneuvers compared with isolated HF.

Sequence analysis of the internal transcribed spacer 2 (ITS2) region of rDNA for identifying Trichogramma species and evaluating genetic diversity

Abstract Species of Trichogramma Westwood, 1833 (Hymenoptera: Trichogrammtidae) are frequently used as biological control agents against Lepidoptera, but practical application of these egg endoparasitoids are complicated because of their complex taxonomy. This study aimed to compare sequences of internal transcribed spacer regions of ribosomal DNA (ITS2-rDNA) of Trichogramma accessions with those deposited in GenBank in order to access the reliability of the ITS2 as a barcode for discriminating species and evaluating the genetic diversity. ITS2-rDNA sequences obtained from seventeen specimens of Trichogramma confirmed previous identifications based on morphological characteristics. Multiple sequence alignment revealed the existence of highly conserved regions in ITS2 sequences while the neighbour-joining dendrogram indicated that the specimens formed three clusters comprising T. manicobai and T. marandobai (group I), T. galloi (group II) and T. pretiosum (group III). The ITS2 marker was shown to be a powerful DNA barcode for discriminating Trichogramma species and could be used to complement the morphological approach.

Resumo Espécies de Trichogramma Westwood, 1833 (Hymenoptera: Trichogrammatidae) são freqüentemente usadas como agentes de controle biológico contra Lepidoptera, esses endoparasitóides de ovos apresentam taxonomia complexa, o que dificulta sua aplicação prática. Este estudo teve como objetivo comparar seqüências de regiões espaçadoras internas transcritas de DNA ribossômico (ITS2-rDNA) de acessos de Trichogramma com aquelas depositadas no GenBank, a fim de avaliar a confiabilidade do ITS2 barcode para discriminar espécies e avaliar a diversidade genética. As seqüências de ITS2-rDNA obtidas de dezessete espécimes de Trichogramma confirmaram identidades anteriores com base em características morfológicas. O alinhamento de múltiplas sequências revelou a existência de regiões altamente conservadas nas sequências ITS2, enquanto o dendrograma indicou que os espécimes formavam três grupos compreendendo T. manicobai e T. marandobai (grupo I), T. galloi (grupo II) e T. pretiosum (grupo III). O marcador ITS2 mostrou ser um poderoso DNA barcode para discriminar espécies de Trichogramma podendo ser usado como complemento da abordagem morfológica.

The influence of fish culture in floating net cages on microbial indicators of water quality / Influencia da piscicultura em tanques-rede sobre os microrganismos indicadores da qualidade de agua

This work was carried out to analyse the microbiological parameters of the water quality of a reservoir used for the irrigation and culture of tilapia (Oreochromis niloticus) in floating net cages. The physico-chemical parameters, counts of mesophilic total aerobic bacteria, total and thermotolerant coliforms and fecal streptococci, and the presence of Escherichia coli in samples of water collected in three sites of the reservoir (pre-culture site, culture site, post-culture site) were analysed. The levels of ammonia (0.047-0.059 mg/L), nitrite (0.001-0.021 mg/L) and total phosphorus (0.050-0.355 mg/L) in the water did not show significant differences (p > 0.05) between sampling sites. The levels of total bacteria in the water varied between 1.3 x 104 and 67.3 x 104 CFU/100 mL. The MPN values of thermotolerant coliforms (< 930 MPN/100 mL) were within values recommended for water used for fish culture and/or irrigation. The presence of E. coli and fecal streptococci were verified in 48% and 56% of analysed samples, respectively. The site with floating net cages showed more samples contaminated with E. coli and fecal streptococci than other sampling points.

Este trabalho foi realizado para analisar os parâmetros microbiológicos da qualidade da água de um reservatório destinado a irrigação e à cultivo de tilapia do Nilo (Oreochromis niloticus) em tanques redes. Foi realizada análise de parâmetros físico-químicos, quantificação de bactérias totais aeróbias mesófilas, coliformes totais e termotolerantes, estreptococos fecais e a detecção de E. coli em amostras de água coletada em três pontos distintos do açude (pré-cultivo, cultivo, pós-cultivo). Os níveis de amônia (0,047-0,059 mg/L), nitrito (0,001-0,021 mg/L) e fósforo total (0,050-0,355 mg/L) na água não apresentaram diferenças significativas (p > 0.05) entre os pontos de coleta. As contagens de bactérias aeróbias mesófilas variaram entre 1,3 x 104 e 67,3 x 104 UFC/100 mL nas amostras de água. O NMP de coliformes termotolerantes (< 930 NMP/100 mL) se apresentou dentro do limite permitido para águas doces destinadas a piscicultura e/ou irrigação. A presença de E. coli foi constatada em 48% e de estreptococos fecais em 56% das amostras de água analisadas. O local de cultivo de tilápia em tanques-rede apresentou maior número de amostras contaminadas com E. coli e estreptococos fecais que outros pontos de amostragem.

Reversal of the anticoagulant and anti-hemostatic effect of low molecular weight heparin by direct prothrombin activation

Lopap, found in the bristles of Lonomia obliqua caterpillar, is the first exogenous prothrombin activator that shows serine protease-like activity, independent of prothrombinase components and unique lipocalin reported to interfere with hemostasis mechanisms. To assess the action of an exogenous prothrombin activator reversing the anticoagulant and antihemostatic effect induced by low molecular weight heparin (LMWH), male New Zealand rabbits (N = 20, weighing 3.8-4.0 kg) allocated to 4 groups were anticoagulated with 1800 IU/kg LMWH (iv) over 2 min, followed by iv administration of saline (SG) or recombinant Lopap (rLopap) at 1 µg/kg (LG1) or 10 µg/kg (LG10), 10 min after the injection of LMWH, in a blind manner. Control animals (CG) were treated only with saline. The action of rLopap was assessed in terms of activated partial thromboplastin time (aPTT), prothrombin fragment F1+2, fibrinogen, and ear puncture bleeding time (BT) at 5, 10, 15, 17, 20, 30, 40, 60, and 90 min after initiation of LMWH infusion. LG10 animals showed a decrease of aPTT in more than 50% and BT near to normal baseline. The level of prothrombin fragment F1+2 measured by ELISA had a 6-fold increase with rLopap treatment (10 µg/kg) and was inversely proportional to BT in LMWH-treated animals. Thus, Lopap, obtained in recombinant form using E. coli expression system, was useful in antagonizing the effect of LMWH through direct prothrombin activation, which can be a possible strategy for the reversal of bleeding and anticoagulant events.

Craniocervical posture analysis in patients with temporomandibular disorder / Análise da postura cranio-cervical em pacientes com disfunção temporomandibular

OBJECTIVE: To compare head positioning and cervical spine alignment between individuals with and without temporomandibular disorders (TMDs), by means of positional evaluation using photographs, radiographs and visual observation, and to investigate whether the type of TMD influences head posture and cervical spine positioning. METHODS: Ninety randomly chosen women were diagnosed using the research diagnostic criteria for TMDs (RDC/TMD) by a trained examiner and were divided into three groups: Group 1, with a diagnosis of myofascial dysfunction (group I of RDC axis I); Group 2, with mixed TMD (groups I, II and III of RDC axis I); and Control, without TMD. Following this, the participants were photographed in frontal and lateral views by a single examiner. To produce these photos, the following anatomical points were marked out on the skin: occipital protuberance, C4, C7, acromioclavicular joint and sternoclavicular joint. From these points, different angles were analyzed by means of the ALCimagem-2000 application. These same photos were then evaluated qualitatively (visual evaluation). Next, lateral teleradiography and radiography of the cervical spine was requested. The examiner was blind when analyzing the images. To compare the results, the chi-squared test and analysis of variance were used, with significance levels of 5 percent. RESULTS: Regardless of the method used, the results revealed that head and cervical spine posture did not differ between the groups with and without TMD, independent of the diagnostic group. CONCLUSION: The posture of individuals with myogenic or arthrogenous TMD does not differ from the posture of individuals without TMD. The presence of TMD does not influence the head and cervical spine posture.

OBJETIVO: Comparar o posicionamento da cabeça e o alinhamento da coluna cervical em indivíduos com e sem DTM, por meio da avaliação postural por fotografias, radiografias e por observação visual e verificar se o tipo de DTM influencia na postura da cabeça e no posicionamento da coluna cervical. MÉTODOS: Noventa mulheres escolhidas aleatoriamente foram diagnosticadas por meio dos Critérios para Diagnóstico em Pesquisa para Disfunções Temporomandibulares (RDC/TMD) por um examinador treinado e divididas em três grupos: grupo 1, diagnóstico de disfunção miofascial (grupo I do eixo I do RDC); grupo 2, com DTM mista (grupo I, II e III do eixo I do RDC) e controle, sem DTM . Em seguida, foram fotografadas em vista anterior e em perfil por um único examinador. Para a realização dessas fotografias, foram demarcados sobre a pele os seguintes pontos anatômicos: protuberância occiptal, C4, C7, articulação acromioclavicular e esternoclavicular. A partir desses pontos, foram analisados diferentes ângulos por meio do aplicativo ALCimagem-2000. Essas mesmas fotos foram posteriormente avaliadas qualitativamente (avaliação visual). Em seguida, foi solicitada uma radiografia e uma telerradiografia em perfil. O examinador foi cego ao analisar as imagens. Para comparação dos resultados, foi utilizado o teste qui-quadrado e a análise de variância com nível de significância de 5 por cento. RESULTADOS: Independentemente do método utilizado, os resultados revelaram que a postura da cabeça e da coluna cervical não diferem entre o grupo com DTM e sem DTM, independentemente do grupo diagnosticado. CONCLUSÃO: A postura do indivíduo com DTM miogênica ou artrogênica não é diferente do indivíduo sem DTM. A presença da DTM não influencia na postura da cabeça e da coluna cervical.

Biochemical and biological properties of Lonomia obliqua bristle extract

Lonomia obliqua caterpillar is frequently seen in accidents with humans especially in the south of Brazil. Patients develop a hemorrhagic syndrome that can be treated with specific antilonomic serum. A consumptive coagulopathy was found to be the main cause of bleeding complications observed in patients after contact with L. obliqua. Studies revealed that L. obliqua caterpillar bristle extract (LOCBE) displays a procoagulant activity that leads to intravascular thrombin formation, resulting in a special form of disseminated intravascular coagulation (DIC). Fibrinolysis seems to be secondary to the fibrin production, since no direct fibrinolytic activity was found in LOCBE. Two procoagulant toxins, a factor X activator (Losac) and a prothrombin activator (Lopap), were isolated from LOCBE and characterized. Infusion of Lopap into experimental animals triggered a condition similar to that observed in human envenomation

A fucan from the brown seaweed Spatoglossum schröederi inhibits Chinese hamster ovary cell adhesion to several extracellular matrix proteins

Fucans, a family of sulfated polysaccharides present in brown seaweed, have several biological activities. Their use as drugs would offer the advantage of no potential risk of contamination with viruses or particles such as prions. A fucan prepared from Spatoglossum schröederi was tested as a possible inhibitor of cell-matrix interactions using wild-type Chinese hamster ovary cells (CHO-K1) and the mutant type deficient in xylosyltransferase (CHO-745). The effect of this polymer on adhesion properties with specific extracellular matrix components was studied using several matrix proteins as substrates for cell attachment. Treatment with the polymer inhibited the adhesion of fibronectin to both CHO-K1 (2 x 10(5))()and CHO-745 (2 x 10(5) and 5 x 10(5)) cells. No effect was detected with laminin, using the two cell types. On the other hand, adhesion to vitronectin was inhibited in CHO-K1 cells and adhesion to type I collagen was inhibited in CHO-745 cells. In spite of this inhibition, the fucan did not affect either cell proliferation or cell cycle. These results demonstrate that this polymer is a new anti-adhesive compound with potential pharmacological applications

Compressão de sinais eletrocardiográficos baseada em transformada discreta do cosseno e codificação de entropia / Compression of electrocardiographic signals based on discrete cosine transform and entropy coding

O presente trabalho relata uma técnica, baseada no padrão JPEG, que combina a Transformada Discreta do Cosseno com técnicas genéricas de compressão de dados visando aumentar a taxa de compressão do sinal de ECG. Dada à grande eficiência do padrão JPEG, buscou-se aplicar pelo menos parte de suas estratégias de compressão ao caso do ECG. A técnica de compressão proposta (CP) permitiu alcançar boas taxas de compressão, com distorções aceitáveis na morfologia do ECG. A razão de compressão média obtida com a CP seguida pela codificação de Huffman foi de 10,2:1 e com a CP seguida pela codificação LZW foi de 9,4:1, para uma PRD = 5 por cento. Traçados de alguns ECGs são mostrados antes e após o processo de compressão/descompressão, permitindo avaliar visualmente a distorção introduzida pelo processamento.

Teores de ácido cianídrico de nove cultivares de capim-sudäo (Sorghum sudanense) / Cianidric acid levels from nine cultivars of sudan grass (Sorghum sudanense)

Nine cultivars of sudan grass cut with 60 days (flowering stage) were used to check the levels of potential cianidric acid (HCN-p), in fresh material. The levels of HCN-p were non toxic and so, there is no risk of toxicity for animals

Incomplete dominance of the low antibody response to Cryptosporidium parvum antigens in mice selected for high and low antibody responsiveness

The humoral antibody response to Cryptosporidium was investigated in mice genetically selected for high (H) and low (L) antibody responsiveness. Groups of 4-5 mice from two different selections, general primary (GP) and general secondary (GS), were studied. Following immunization with Cryptosporidium parvum antigens, the maximum levels of IgG in the HGP, (X ñ SD = 1.13 ñ 0.35, N = 5) and in the HGS (0.42 ñ 0.15, N = 4) lines, and of IgM in the HGP line (0.86 ñ 0.53, N = 5) were significantly higher than those in their L counterparts (0.04 ñ 0.02, N = 5;0.05 ñ 0.02, N = 4 and 0.24 ñ 0.07, N = 5, respectively). These findings were similar to those reported for other immunogens. However, the IgG (0.22 ñ 0.05, N = 4) and the IgM (0.33 ñ 0.08, N = 4) responses to immunization of F1 (LGP x HGP) hybrids indicated an incomplete dominance of the low response, in contrast to the incomplete dominance of the high response described for many other antigens and representing an important exception. In addition, the H, L and F1 mice did not develop detectable infections when inoculated with live Cryptosporidium oocysts, supporting the view that a reduced or zero antibody production itself is not enough to permit the establishment of Cryptosporidium infection in adult mice.

A study on the amphiphilic proteins of three Trypanosoma cruzi populations

Amphiphilic proteins were partially purified from culture-derived metacyclic trypomastigotes of the CL and Colombian strains and of the Dm 28c clone of T. cruzi by the use of Triton X-114. These proteins were subjected to one-and two-dimensional polyacrylamide electrophoresis in the presence of sodium dodecyl sulphate. Relatively simple protein profiles with only 5 to 6 major bands were obtained. The CL and Colombian strains produced at least one additional major protein band (86 kDa) compared to the Dm 28c proteins. Trypomastigote amphiphilic proteins displayed both electrophoretic mobilities and isoelectric points identical to those of two polypeptides precipitated by a rabbit antiserum which recognized metacyclic trypomastigote-specific surface antigens. The partial purification of the T. cruzi amphiphilic proteins with Triton X-114 may provide a simple preparative step for the study of these differentiation-related polypeptides, as well as for the study of strain-specific (glyco)proteins and of their possible biological role

Restriction in the repertoire of detectable autoantibodies in polyclonal B cell activations and the mimicry of autoantigens by idiotopes

Despite the existence of erythrocyte-autoreactive B cells in normal animals, erythrocyte-autoantibodies could not be detected during polyclonal B-cell activation (PBA) both in patients with visceral leishmaniasis and in bacterial lipopolysacharide (LPS) - injected mice. The failure to detect these autoantibodies in mice with PBA di not seem to be due to suppressor-cell activity, since (1) transfer of spleen cells from LPS-treated mice to naive recipients did not affect the erythrocyte-autoantibody response elicited by subsequent injections of rat erythrocytes and (2) low doses of X-radiation did no lead to erythrocyte-autoantibody detection in LPS-treated mice. The possibility that the detection of erytrocyte-autoantibodies could be affected by autoantibodies with idiotopes mimicring erythrocyte epitopes, the synthesis of which would also be triggerred in PBA, is discussed. Indirect evidence for the existence in normal animal of an expanded lymphocyte population with DNP-binding. Ia-mimicring antigen receptors is presented.