Cloning and characterization of SmZF1, a gene encoding a Schistosoma mansoni zinc finger protein

The zinc finger motifs (Cys2His2) are found in several proteins playing a role in the regulation of transcripton. SmZF1, a Schistosoma mansoni gene encoding a zinc finger protein was initially isolated from an adult worm cDNA library, as a partial cDNA. The full sequence of the gene was obtained by subcloning and sequencing cDNA and genomic fragments. The collated gene sequence is 2181 nt and the complete cDNA sequence is 705 bp containing the full open reading frame of the gene. Analysis of the genome sequence revealed the presence of three introns interrupting the coding region. The open reading frame theoretically encodes a protein of 164 amino acids, with a calculated molecular mass of 18,667Da. The predicted protein contains three zinc finger motifs, usually present in transcription regulatory proteins. PCR amplification with specific primers for the gene allowed for the detection of the target in egg, cercariae, schistosomulum and adult worm cDNA libraries indicating the expression of the mRNA in these life cycle stages of S. mansoni. This pattern of expression suggests the gene plays a role in vital functions of different life cycle stages of the parasite. Future research will be directed to elucidate the functional role of SmZF1.

The zinc finger protein TcZFP2 binds target mRNAs enriched during Trypanosoma cruzi metacyclogenesis

Trypanosomes are parasitic protozoa in which gene expression is primarily controlled through the regulation of mRNA stability and translation. This post-transcriptional control is mediated by various families of RNA-binding proteins, including those with zinc finger CCCH motifs. CCCH zinc finger proteins have been shown to be essential to differentiation events in trypanosomatid parasites. Here, we functionally characterise TcZFP2 as a predicted post-transcriptional regulator of differentiation in Trypanosoma cruzi. This protein was detected in cell culture-derived amastigotes and trypomastigotes, but it was present in smaller amounts in metacyclic trypomastigote forms of T. cruzi. We use an optimised recombinant RNA immunopreciptation followed by microarray analysis assay to identify TcZFP2 target mRNAs. We further demonstrate that TcZFP2 binds an A-rich sequence in which the adenosine residue repeats are essential for high-affinity recognition. An analysis of the expression profiles of the genes encoding the TcZFP2-associated mRNAs throughout the parasite life cycle by microarray hybridisation showed that most of the associated mRNAs were upregulated in the metacyclic trypomastigote forms, also suggesting a role for TcZFP2 in metacyclic trypomastigote differentiation. Knockdown of the orthologous Trypanosoma brucei protein levels showed ZFP2 to be a positive regulator of specific target mRNA abundance.

Cyanogenesis and the onset of tapping panel dryness in rubber tree

The objective of this work was to study the influence of cyanogenesis on the onset of irreversible tapping panel dryness (TPD) and the physiological and histological aspects of secondary phloem in the trunk (tapping panel) of rubber trees (Hevea spp.). Two cyanogenic compounds, linamarin and KCN, were applied separately on the trunk bark of healthy mature trees belonging to two Brazilian clones (Fx 4098 and Fx 3899). Changes in histology, latex pressure potential (ΨP) and cyanogenic potential (HCNp) were followed in the trunk inner barks. In addition, the HCNp levels were determined in TPD-affected plants of both clones. The applications of linamarin or KCN in healthy plants decreased latex ΨP, and formed tylosoids associated with in situ coagulation of latex. The clone Fx 4098 had the higher HCNp and showed the quicker and stronger responses to the cyanogenic compounds. Plants with TPD syntoms had a higher HCNp than the untreated healthy ones. Since histological changes are also structural markers of early TPD, it can be inferred that excessive release of cyanide can induce it in sensitive rubber clones

In silico analysis identifies a C3HC4-RING finger domain of a putative E3 ubiquitin-protein ligase located at the C-terminus of a polyglutamine-containing protein

Almost identical polyglutamine-containing proteins with unknown structures have been found in human, mouse and rat genomes (GenBank AJ277365, AF525300, AY879229). We infer that an identical new gene (RING) finger domain of real interest is located in each C-terminal segment. A three-dimensional (3-D) model was generated by remote homology modeling and the functional implications are discussed. The model consists of 65 residues from terminal position 707 to 772 of the human protein with a total length of 796 residues. The 3-D model predicts a ubiquitin-protein ligase (E3) as a binding site for ubiquitin-conjugating enzyme (E2). Both enzymes are part of the ubiquitin pathway to label unwanted proteins for subsequent enzymatic degradation. The molecular contact specificities are suggested for both the substrate recognition and the residues at the possible E2-binding surface. The predicted structure, of a ubiquitin-protein ligase (E3, enzyme class number 6.3.2.19, CATH code 3.30.40.10.4) may contribute to explain the process of ubiquitination. The 3-D model supports the idea of a C3HC4-RING finger with a partially new pattern. The putative E2-binding site is formed by a shallow hydrophobic groove on the surface adjacent to the helix and one zinc finger (L722, C739, P740, P741, R744). Solvent-exposed hydrophobic amino acids lie around both zinc fingers (I717, L722, F738, or P765, L766, V767, V733, P734). The 3-D structure was deposited in the protein databank theoretical model repository (2B9G, RCSB Protein Data Bank, NJ).

Myocardial ischemic preconditioning upregulated protein 1(Mipu1):zinc finger protein 667 - a multifunctional KRAB/C2H2 zinc finger protein

Myocardial ischemic preconditioning upregulated protein 1 (Mipu1) is a newly discovered upregulated gene produced in rats during the myocardial ischemic preconditioning process. Mipu1 cDNA contains a 1824-base pair open reading frame and encodes a 608 amino acid protein with an N-terminal Krüppel-associated box (KRAB) domain and classical zinc finger C2H2 motifs in the C-terminus. Mipu1 protein is located in the cell nucleus. Recent studies found that Mipu1 has a protective effect on the ischemia-reperfusion injury of heart, brain, and other organs. As a nuclear factor, Mipu1 may perform its protective function through directly transcribing and repressing the expression of proapoptotic genes to repress cell apoptosis. In addition, Mipu1 also plays an important role in regulating the gene expression of downstream inflammatory mediators by inhibiting the activation of activator protein-1 and serum response element.

Aspectos fisiológicos e clínicos da técnica fonoterapêutica de fonação reversa

A fonação reversa é a produção de voz durante a inspiração, realizada espontaneamente em situações como o suspiro. OBJETIVO: Realizar uma revisão da literatura, descrevendo achados relacionados à utilização da fonação reversa na prática clínica, à anatomofisiologia de sua produção e seus efeitos no trato vocal, e às indicações e contra-indicações da técnica para os distúrbios e para o aperfeiçoamento da voz. RESULTADOS: Foram encontrados relatos de mudanças significativas no trato vocal durante a produção da fonação reversa, como o relaxamento dos ventrículos, o afastamento das pregas ventriculares, o aumento da freqüência fundamental, o movimento inverso da onda mucosa, além da facilitação do estudo dinâmico da laringe, quando associada à endoscopia, o que possibilita melhor definição da localização das lesões nas camadas da lâmina própria da mucosa das pregas vocais. CONCLUSÃO: Existem poucos estudos que descrevem o comportamento laríngeo durante a fonação reversa e, para que essa técnica utilizada de forma mais precisa e objetiva, acredita-se que ainda devem ser realizados estudos que visem a comprovar sua eficácia na prática clínica.

Qualidade da cebola influenciada pelo estádio de maturação da planta na colheita

O objetivo deste trabalho foi avaliar a qualidade dos bulbos de duas variedades de cebola colhidos em três estádios de maturação da planta. Bulbos das variedades Texas Grano e Baia Periforme foram colhidos em 2006 no campo experimental da Universidade Federal de Viçosa, de plantas com 10% de estalo natural, 10% de estalo induzido e 50% de estalo natural. Na colheita foram avaliados a massa da matéria fresca total, o diâmetro, a cor (L*, a* e b*), o teor de massa da matéria seca, os sólidos solúveis totais e o teor relativo de água da polpa e das películas externas do bulbo. No cultivar Texas Grano, a massa da matéria fresca total, o diâmetro e os teores de sólidos solúveis totais foram maiores nos bulbos colhidos com 10 e 50% de estalo natural. O teor relativo de água das películas externas de bulbos variedade Texas Grano foi maior somente quando colhidos com 10% de estalo natural, o que representa bulbos com películas mais úmidas. Bulbos colhidos com 10% de estalo natural e 10% de estalo induzido tiveram valores de L* maiores do que aqueles colhidos com 50% de estalo natural, indicando que a antecipação da colheita reduz a intensidade da cor das películas. O menor valor de b* para a película do bulbo da Texas Grano, colhido com 10% estalo natural, decorre do amarelo menos intenso da casca. No cultivar Baia Periforme, os índices de maturação não influenciaram a massa da matéria fresca total, o diâmetro, teor de massa da matéria seca, o teor de sólidos solúveis totais e os índices de cor a* e b*. O estádio de maturação da planta, na colheita, não influencia o tamanho e o teor de sólidos solúveis dos bulbos na variedade Baia Periforme. Para a Texas Grano, bulbos maiores, com películas mais amareladas e menor teor de umidade são obtidos quando a colheita é realizada com 50% de estalo natural.

Biochemical and morpho-anatomical analyses of strawberry vitroplants hyperhydric tissues affected by BA and gelling agents

In vitro propagation has become an effective practice for large-scale production of strawberry plants. The objective of this study was to evaluate the hyperhydricity and the multiplication capacity of two strawberry varieties (Fragaria x ananassa Duch. 'Dover' and 'Burkley') propagated in vitro. Plants maintained in MS medium supplemented with 1.0 mg L-1 BA were individualized and transferred to the same medium solidified with Agar (6.5 g L-1) or Phytagel® (2.5 g L-1) and BA at different concentrations (0; 0.5; 1.0; 2.0 and 3.0 mg L-1). Biochemical and anatomical analyses were carried out, as well as the analysis of the morphological hyperhydricity characteristics. The analysis of data showed: a) the increase in cytokinin concentration increased hyperhydricity frequency in both varieties; b) at concentrations up to 2.0 mg L-1 BA, the replacement of Agar by Phytagel® induced a higher formation of hyperhydric shoots; and c) the addition of BA induced oxidative stress, which is characterized by increased antioxidant activity and lipid peroxidation, as well as alterations at the cellular level, such as malformation of stomata and epidermal cells. In conclusion, the culture medium containing 0.5 mg L-1 BA solidified with Agar provided lower hyperhydricity percentages in association with higher rates of shoot proliferation in strawberry.

Carbohydrates Concentration in leaves of potato plants affected by nitrogen fertilization rates

ABSTRACT At poor conditions of nitrogen (N) in the soil, potato plants may accumulate starch in leaves and be indicative of N nutritional stress. The objective of this work was to determine the effects of N rates (0, 50, 100, 200 and 300 kg ha-1 of N) on the concentrations of carbohydrates (total soluble sugars-TSS, reducing sugars-RS, non-reducing sugars-NRS and starch) in the fourth leaf (FL) of two potato cultivars (Asterix and Atlantic) and their critical levels (CL) associated to the N fertilization rate necessary to obtain the maximum physical (MPE) and economic (MEE) efficiency of tubers. A randomized block design with four replications was used in both experiments. On day 21 after plant emergence, four FL were collected from four plants. Potatoes plants fertilized with low rates of N accumulated less TSS in leaves than those properly fertilized. The opposite occurred with content of starch. The cultivars showed similar responses to five doses of N in relation to contents of starch and TSS. However, the response to the increase in doses of N for RS, NRS and Starch/NRS is cultivar-specific. The correlations between contents of RS, NRS and Starch/NRS with the starch and TSS were dependent on the potato cultivar.

Novas tecnologias e desempenho operacional: um estudo internacional comparativo

Este estudo investiga a influência da antecipação de novas tecnologias sobre o desempenho em empresas de manufatura. Complementarmente, foi realizada uma análise comparativa entre plantas industriais localizadas em outros países e no Brasil. Para testar essas questões, foram coletados dados de 337 empresas em 11 países, incluindo o Brasil. Aspectos-chave na gestão de operações, como estratégia de manufatura, integração funcional, gestão da cadeia de suprimentos e parceria com fornecedores, foram considerados relevantes para antecipação de novas tecnologias por empresas. Os resultados sugerem que estratégia de manufatura, integração funcional e parceria com fornecedores são importantes para a antecipação de novas tecnologias por empresas localizadas em outros países, enquanto apenas a integração funcional é importante para as empresas brasileiras. Por fim, existem evidências que sugerem um impacto positivo entre antecipação de novas tecnologias e desempenho operacional, tanto para as empresas internacionais como para as empresas brasileiras.

Assessment of oltipraz in schistosomiasis mansoni clinical trial

Seventy three children (6-15 years) and 75 adults (18-47 years) with active schistosomiasis mansoni were treated with oltipraz. All cases had at least 100 eggs per gram of feces as determined by the Kato-Katz technique. Children and adults were divided in two groups receiving respectively 25 or 30 mg/kg, as a single oral dose. Clinical examination, laboratories tests (haemogram, urinalysis, hepatic and kidney functions tests, glycemia, cholesterol, triglicerides, lipoprotein — HLD and LDL) and ECG were performed before, 3 or 7 days and 1 month after treatment. Parasitological control with 3 daily coprological examinations, was done on the 1st, 3rd j 6th month after drug administration. Giddiness, somnolence, headache, nausea, vomiting and abdominal distress were the most frequent side effects. Pain in the finger tips that need further investigations also occurred. No significant alteration in complementary tests were observed, whereas eosinophilia 1 month after treatment was detected, probably indicating worm death. The cure rate in children was 81.8% and 74.2% with 25 and 30 mg/kg respectively, and in adults 75.0% and 81.2% of the patients. No statistical significant difference was observed between cure rate and side effects at different dosages employed, neither between adults nor children. In all groups the percentage of egg reduction in feces in the non cured patients was higher than 96.0%. Further investigation with this new compound is necessary to accomplish the real value of oltipraz in the schistosomiasis chemotherapy.

Subcutaneous phaeohyphomycosis caused by Bipolaris hawaiiensis: a case report

A case of phaeohyphomycosis caused by Bipolaris hawaiiensis is reported. The patient, an immunocompetent host, presented a verrucous lesion on the first finger of the left foot. Dematiaceous septate hyphae and yeast-like elements were seen in direct and histological examination. The isolated strain was identified on the basis of micro and macromorphological aspects. Treated with electrocoagulation, the lesions healed and presented no relapse after two years follow-up.

Anti-phenolic glycolipid-I (PGL-I) determination using blood collection on filter paper in leprosy patients

The authors studied 70 leprosy patients and 20 normal individuals, comparing the traditional sera collection method and the finger prick blood with the conservation on filter paper for specific antibodies against the native phenolic glycolipid-I (PGL-I) from Mycobacterium leprae. The finger prick blood dried on filter paper was eluated in phosphate buffer saline (PBS) containing 0.5% gelatin. The classical method for native PGL-I was performed for these eluates, and compared with the antibody determination for sera. It was observed that there is a straight correlation comparing these two methods; although the titles found for the eluates were lower than those obtained for serology. This blood collection method could be useful for investigation of new leprosy cases in field, specially in contacts individuals.

Envenomation by neotropical Opisthoglyphous colubrid Thamnodynastes cf. pallidus Linné, 1758 (Serpentes:Colubridae) in Venezuela

This is a case report of a "non-venomous" snake bite in a herpetologist observed at the Sciences Faculty of the Universidad de los Andes (Mérida, Venezuela). The patient was bitten on the middle finger of the left hand, and shows signs of pronounced local manifestations of envenomation such as bleeding from the tooth imprint, swelling and warmth. He was treated with local care, analgesics, and steroids. He was dismissed from the hospital and observed at home during five days with marked improvement of envenomation. The snake was brought to the medical consult and identified as a Thamnodynastes cf. pallidus specimen. This report represents the first T. pallidus accident described in a human.

SEROLOGICAL AND MOLECULAR SURVEY OF Leptospira spp. AMONG CART HORSES FROM AN ENDEMIC AREA OF HUMAN LEPTOSPIROSIS IN CURITIBA, SOUTHERN BRAZIL

Introduction: Cart horses are a re-emerging population employed to carry recyclable material in cities. Methods: Sixty-two horses were sampled in an endemic area of human leptospirosis. The microscopic agglutination test (MAT) and real-time polymerase chain reaction (qPCR) were performed. Results: A seropositivity of 75.8% with serovar Icterohaemorrhagiae in 80.8% of the horses was observed. Blood and urine were qPCR negative. MAT showed positive correlations with rainfall (p = 0.02) and flooding (p = 0.03). Conclusions: Although horses may be constantly exposed to Leptospira spp. in the environment mostly because of rainfall and flooding, no leptospiremia or leptospiruria were observed in this study.