The Shiga toxin 2 B subunit inhibits net fluid absorption in human colon and elicits fluid accumulation in rat colon loops

Shiga toxin (Stx)-producing Escherichia coli (STEC) colonizes the large intestine causing a spectrum of disorders, including watery diarrhea, bloody diarrhea (hemorrhagic colitis), and hemolytic-uremic syndrome. It is estimated that hemolytic-uremic syndrome is the most common cause of acute renal failure in infants in Argentina. Stx is a multimeric toxin composed of one A subunit and five B subunits. In this study we demonstrate that the Stx2 B subunit inhibits the water absorption (Jw) across the human and rat colonic mucosa without altering the electrical parameters measured as transepithelial potential difference and short circuit current. The time-course Jw inhibition by 400 ng/ml purified Stx2 B subunit was similar to that obtained using 12 ng/ml Stx2 holotoxin suggesting that both, A and B subunits of Stx2 contributed to inhibit the Jw. Moreover, non-hemorrhagic fluid accumulation was observed in rat colon loops after 16 h of treatment with 3 and 30 ng/ml Stx2 B subunit. These changes indicate that Stx2 B subunit induces fluid accumulation independently of A subunit activity by altering the usual balance of intestinal absorption and secretion toward net secretion. In conclusion, our results suggest that the Stx2 B subunit, which is non-toxic for Vero cells, may contribute to the watery diarrhea observed in STEC infection. Further studies will be necessary to determine whether the toxicity of Stx2 B subunit may have pathogenic consequences when it is used as a component in an acellular STEC vaccine or as a vector in cancer vaccines.

Primary mechanism of apoptosis induction in a leukemia cell line by fraction FA-2-b-ß prepared from the mushroom Agaricus blazei Murill

Agaricus blazei Murill is a native Brazilian mushroom which functions primarily as an anticancer substance in transplanted mouse tumors. However, the mechanism underlying this function of A. blazei Murill remains obscure. The present study was carried out to investigate the effect of fraction FA-2-b-ß, an RNA-protein complex isolated from A. blazei Murill, on human leukemia HL-60 cells in vitro. Typical apoptotic characteristics were determined by morphological methods using DNA agarose gel electrophoresis and flow cytometry. The growth suppressive effect of fraction FA-2-b-ß on HL-60 cells in vitro occurred in a dose- (5-80 µg/mL) and time-dependent (24-96 h) manner. The proliferation of HL-60 cells (1 x 10(5) cells/mL) treated with 40 µg/mL of fraction FA-2-b-ß for 24-96 h and with 5-80 µg/mL for 96 h resulted in inhibitory rates ranging from 8 to 54.5%, and from 4.9 to 86.3%, respectively. Both telomerase activity determined by TRAP-ELISA and mRNA expression of the caspase-3 gene detected by RT-PCR were increased in HL-60 cells during fraction FA-2-b-ß treatment. The rate of apoptosis correlated negatively with the decrease of telomerase activity (r = 0.926, P < 0.05), but correlated positively with caspase-3 mRNA expression (r = 0.926, P < 0.05). These data show that fraction FA-2-b-ß can induce HL-60 cell apoptosis and that the combined effect of down-regulation of telomerase activity and up-regulation of mRNA expression of the caspase-3 gene could be the primary mechanism of induction of apoptosis. These findings provide strong evidence that fraction FA-2-b-ß could be of interest for the clinical treatment of acute leukemia.

Síntese e caracterização de cloro[ácido 2-(difenilfosfino)benzóico]-ouro(I) e alguns de seus derivados

In this contribution a few new gold(I)phosphine complexes, [2-(PPh2)C6H4CO 2H]AuX (where X = Cl, SCN, Br3) and a similar gold(III) derivative [{2-(PPh2)C6H4CO 2H}AuIII Cl (C6H4CH2NMe2 )]Cl have been synthesised and characterised. The phosphine, 2-(diphenylphosphino)benzoic acid, has been employed for the first time in gold chemistry. This ligand is potentially bidentate through bonding of the phosphine and carboxylate groups. The X-ray structure of the complex chloro[2-(diphenylphosphino) benzoic acid]gold(I) has been elucidated and the bond lengths encountered show great similarity to those of chloro(triphenylphosphine)gold(I). [2-(PPh2)C6H4CO 2H]AuCl crystallises in the space group P2(1)/c with a = 9.113(2) Å, b = 10.925(2) Å, c = 23.069(4) Å, beta = 99.95º(3), V = 2299 ų, Z = 4 and R = 0.091. Biological tests for anti-fungal and anti-bacterial activity demostrate that [2-(PPh2)C6H4CO 2H]AuCl exhibits broad spectrum activity against a range of organisms.

Acidente por serpentes do gênero Bothrops: série de 3.139 casos

Em avaliação dos prontuários médicos de 3.139 pacientes picados por serpentes do gênero Bothrops atendidos no Hospital Vital Brazil (HVB), de 1981 a 1990, observou-se maior acometimento do sexo masculino (75,7%). Em 1.412 casos (45,0%) a serpente foi identificada, sendo 1.376 B. jararaca, 20 B. jararacussu, 11 B. neuwiedi, 2 B. moojeni, 2 B. alternatus e 1 B. pradoi. As regiões anatômicas mais comumente picadas foram: pé (47,5%) e mão (21,3%). O torniquete foi realizado em 38,2% dos casos e sua freqüência diminuiu durante esse período (p < 0,001). As manifestações clínicas mais freqüentes foram: dor (95,6%), edema (95,4%), equimose (56,1%), bolha (13,8%), necrose (16,5%), abscesso (11,0%), sangramento extratecidual (12,3%), insuficiência renal (1,6%) e choque (0,7%). A coagulação sanguínea foi avaliada em 2.990 casos e estava alterada em 1.730 (57,9%). Ocorreram 21 amputações (0,7%) e 9 óbitos (0,3%). A dose de soro administrada no HVB diminuiu ao longo desse período (p < 0,001).

A survey of planorbid molluscs in the Amazonian region of Brazil

A survey of the plarnorbid fauna in the Brazilian states of the Amazonian river basin revealed the occurence of 14 species, 8 of the genus Biomphalaria, 4 of Drepanotrema, 1 of Antillorbis and 1 of Plesiophysa, besides a naturalized puopulation of Helisoma duryi at Santa Rosa, municipality of Formosa, state of Goiás. The following is the distribution of the species by genera, in decreasing order of frequency (number of localities in parenthesis): 1. Biomphalaria straminea (50): Acre, Amazonas, Distrito Federal, Goiás, Maranhão, Mato Grosso, Mato Grosso do Sul, Pará and Roraima; 2. B. occidentalis (30): Acre, Amazonas, Mato Grosso and Mato Grosso do Sul; 3. B. schrammi (22); Distrito Federal, Goiás, Maranhão, Mato Grosso, Mato Grosso do Sul and Pará; 4. B. amazonica (14): Acre, Amazonas and Rondônia; 5. B. glabrata (13): Distrito Federal, Goiás, Maranhão and Pará; 6. B. peregrina (4): Distrito Federal, Goiás and Mato Grosso do Sul; 7. B. tenagophila (2): Distrito Federal and Goiás; 8. B. oligoza (2): Mato Grosso do Sul; 9. Drepanotrema lucidium (72): Acre, Amapá, Amazonas, Distrito Federal, Goiás, Maranhão, Mato Grosso, Mato Grosso do Sul, Pará, Rondônia and Roraima; 10. D. anatinum (41): Acre, Amazonas, Distrito Federal, Goiás, Maranhão, Mato Grosso do Sul, Pará, Rondônia and Roraima; 11. D. depressissimum (19): Acre, Amazonas, Distrito Federal, Goiás, Maranhão, Mato Grosso, Mato Grosso do Sul and Pará; 12. D. cimex (15): Acre, Amazonas, Distrito Federal, Goiás, Mato Grosso, Mato Grosso do Sul and Pará; 13. Antillorbis nordestensis (3): Distrito Federal, Maranhão and Pará; 14. Plesiophysa ornata (1): Goiás. B. glabrata is responsibel for transmission of schistosomiasis mansoni in northeastern Pará, northern Marnhão and central Goiás including the Distrito Fedreal. B. tenagophila, although susceptible to experimental infection with Schistosoma mansoni, has not been found naturally infected so far in the area. B. straminea has been incriminated as...

Geographical distribution of Biomphalaria snails in the State of Minas Gerais, Brazil

Published and unpublished observations on geographical distribution of Biomphalaria snails in the State of Minas Gerais, Brazil, were compiled. This work is aimed at knowing the present occurrence of Biomphalaria species in this region, and at contributing to the elaboration of the planorbid chart of Minas Gerais. In malacological surveys, performed by several researchers, the presence of seven species of this genus was recorded. Those planorbids were found in 12 mesoregions, in 283 (33.1%) municipalities out of 853 with the following distribution: B. glabrata (185 municipalities), B. straminea (125), B. tenagophila (58), B. peregrina (57), B. schrammi (26), B. intermedia (20) and B. occidentalis (2). B. glabrata and B. tenagophila are found naturally infected by Schistosoma mansoni in Minas Gerais. In 24 municipalities the three snail hosts of S. mansoni in Brazil, B. glabrata, B. tenagophila and B. straminea, are present.

Interaction between the Intermediate Host of Schistosomiasis in Brazil Biomphalaria glabrata (Planorbidae) and a Possible Competitor Melanoides tuberculata (Thiaridae): I. Laboratory Experiments

The biological control of Biomphalaria glabrata, intermediate host of Schistosoma mansoni, is one the accepted options to fight schistosomiasis. One of the most promising candidates to control B. glabrata is the snail Melanoides tuberculata, a potential competitor. However, the mechanisms of interaction between the two species are not clear. Our objective is to determine if M. tuberculata indeed compete with B. glabrata, using two laboratory experiments. In Experiment 1, we tested the effect of the presence of M. tuberculata on the fecundity and mortality rates of B. glabrata. In Experiment 2, we tested if there was a direct or indirect interaction between the two species. In Experiment 1, M. tuberculata was eliminated after the peak in reproductive activity of B. glabrata. In Experiment 2, B. glabrata produced more egg masses when raised with M. tuberculata. The conditions leading to this unexpected positive effect of M. tuberculata on the fecundity of B. glabrata need further clarification, but emphasize that detailed studies of the interaction between these species in the conditions of the local environment should be considered.

Virulence markers and antimicrobial susceptibility of bacteria of the Bacteroides fragilis group isolated from stool of children with diarrhea in São Paulo, Brazil

Bacteroides fragilis has been isolated from several human and non-human monomicrobial and mixed infections. In this study, some virulence markers and the antimicrobial susceptibility of bacteria of the B. fragilis group isolated from children's stools were evaluated. All the 64 isolates showed the following characteristics: capsulated, beta-hemolytic, hydrophilic, and serum-resistant. Only, 24 (37.5%) strains were resistant at 60ºC, for 30 min, and among them, 12 (18.75%) were resistant at 60ºC, for 60 min. Also, none strain was resistant at 100ºC. Four strains were able to hemagglutinate erythrocytes and D-mannose, D-galactose, D-arabinose, and D-xylose inhibited hemagglutination in 2 B. fragilis strains (p76a, p76b). The hemagglutination in the strain B. uniformis p3-2 was inhibited by D-xylose and D-galactose. The bft gene detection and the enterotoxin production were observed only in 13 EF-enterotoxigenic species. Fragilysin activity was confirmed on HT-29 cells. The antimicrobial determination confirmed that both imipenem and metronidazole were efficient against B. fragilis species; all the strains were resistant to lead and nickel. Plasmids of 2.9, 4.4, 4.8, and 8.9 kb were observed in 6 tested strains. These results show the values of the species identification from clinical infections, as well as of the periodic evaluation of the resistance patterns of the B. fragilis group at Brazilian medical institutions.

Physiological parameters to select upland rice genotypes for tolerance to water deficit

Abstract: The objective of this work was to evaluate the feasibility of using physiological parameters for water deficit tolerance, as an auxiliary method for selection of upland rice genotypes. Two experiments - with or without water deficit - were carried out in Porangatu, in the state of Goiás, Brazil; the water deficit experiment received about half of irrigation that was applied to the well-watered experiment. Four genotypes with different tolerance levels to water stress were evaluated. The UPLRI 7, B6144F-MR-6-0-0, and IR80312-6-B-3-2-B genotypes, under water stress conditions, during the day, showed lower stomatal diffusive resistance, higher leaf water potential, and lower leaf temperature than the control. These genotypes showed the highest grain yields under water stress conditions, which were 534, 601, and 636 kg ha-1, respectively, and did not differ significantly among them. They also showed lower drought susceptibility index than the other genotypes. 'BRS Soberana' (susceptible control) was totally unproductive under drought conditions. Leaf temperature is a easy-read parameter correlated to plant-water status, viable for selecting rice genotypes for water deficit tolerance.

Síntese e avaliação da atividade fitotóxica de derivados da α-Santonina

Mixtures of α-Santonin and various solvents were irradiated by either high or low pressure mercury lamps. The photochemical reactions afforded lumisantonin (11) (76% in acetonitrile), (3S,3aS,9bS)-3,6,6-trimethyl-3,3a,4,5-tetrahydronafto[1,2-b]furan-2,7(6H,9bH)dione (12) (100% in acetonitrile), 10α-acetoxy-3-oxo-1,7αH,6,11βH-guaia-4-en-6,12-olide (8) (26% in acetic acid), 10α-hydroxy-3-oxo-1,7αH,6,11βH-guaia-4-en-6,12-olide (10) (32%) and (E)-3-((3S,3aS,7aS)-3-methyl-2-oxo-6-(propan-2-ylidene)hexahydrobenzofuran-7-(7aH)-ylidene)propanoic acid (9) (44%) (in water/ acetic acid 1:1, v/v). Lactone 12 was also prepared by irradiation of lumisantonin in diethyl ether. Lactones 8 and 10 were converted, respectively, into the 10α-acetoxy-3β-hydroxy-1,7αH,6,11βH-guaia-4-en-6,12-olide (13) (87%) and 3β,10α-dihydroxy-1,7αH,6,11βH-guaia-4-en-6,12-olide (14) (75%) by sodium borohydride reduction. The effects of the compounds on the development of radicle of Sorghum bicolor and Cucumis sativus were evaluated.

Electrochemical study, on mercury, of a Meta-nitroarylamine derivative of nor-β-lapachone, an antitumor and trypanocidal compound

The electrochemistry of 2,2-dimethyl-(3H)-3-(N-3'-nitrophenylamino)naphtho[1,2- b]furan-4,5-dione ([Q]-PhNO2), on mercury was investigated. The first peak is consistent with a quasi-reversible one-electron reduction of the ortho-quinone, forming [Q•-]-PhNO2, while the second one, bielectronic, corresponds to the simultaneous reduction of the latter radical to a dianion and the nitro group to a nitro radical anion. The second order rate constant, k disp, for the decay of [Q•-]-PhNO2 is 15.188 x 10³ ± 827 mol"1 L s"1 and the t1/2 equals 0.06 s. E¹7Ic values for [Q]-PhNO2 and its precursor, nor-β-lapachone, are similar. The ease of semiquinone generation and its stability are parameters statistically relevant in the correlation biochemical/theoretical aspects.

Avaliação da influência do tempo e do diâmetro na obstrução da janela pleuropericárdica

OBJETIVO: Avaliar a influência do diâmetro e do tempo na obstrução da janela pleuro-pericárdica em cães com pericárdio normal. MÉTODO: Trinta e seis cães mestiços foram divididos em seis grupos: 1 a, 1 b,1 c, 2 a, 2 b, 2 c; n=6 por grupo. Nos grupos 1 a , 1 b, 1 c, à janela foi de 2cm de diâmetro e nos grupos 2 a, 2 b, 2 c, foi de 4cm. Os animais foram reoperados respectivamente após 2, 8 e 12 semanas. Na re-operação, avaliou-se o grau de obstrução através de uma escala de aderência pericárdio-epicárdica e a histopatologia das bordas do pericárdio. RESULTADOS: Numa análise global, observou-se 89% de janelas abertas e 11% de janelas totalmente obstruídas. No grupo com 2cm de diâmetro original, na reoperação, encontrou-se um diâmetro maior em 89% dos cães, enquanto que nos cães com janela original de 4cm, isto ocorreu em 61%. Quando comparou-se os resultados nos cães com janelas de diâmetro igual, mas re-operados em diferentes tempos de pós-operatório, não se observou diferença estatisticamente significante. O mesmo ocorreu quando comparou-se os cães com janelas de diâmetro diferente e re-operados em tempos iguais de pós-operatório. O grau de aderência pericárdio-epicárdica, de acordo com a escala de gradação, não foi diferente entre os vários grupos em função do tempo e diâmetro com exceção do grupo com janela de 4cm e re-operado com 8 semanas (Grupo 2b). As alterações histopatológicas não foram estatisticamente significantes entre os grupos. Em nenhum cão observou-se obstrução da janela pleuropericárdica pelo pulmão. CONCLUSÃO: o tempo e o diâmetro da janela pleuropericardica não influenciaram na obstrução da mesma.

Genetic grouping of avian infectious bronchitis virus isolated in Brazil based on RT-PCR/RFLP analysis of the S1 gene

Twelve Brazilian isolates and one reference vaccine strain of avian infectious bronchitis virus (IBV) were propagated in embryonating chicken eggs. The entire S1 glycoprotein gene of these viruses was analysed by reverse-transcriptase-polymerase chain reaction and restriction fragment length polymorphism (RT-PCR-RFLP), using the restriction enzymes HaeIII, XcmI and BstyI. The RFLP patterns led to the classification of these isolates into five distinct genotypes: A, B, C, D and Massachusetts. Five of twelve isolates were grouped in Massachusetts genotype and the remaining seven viruses were classified into four distinct genotypes: A (2), B (2), C (2) or D (1). Such genotyping classification agreed with previous immunological analysis for most of these viruses, highlighting the occurrence of a relevant variability among the IBV strains that are circulating in Brazilian commercial poultry flocks.

Competição relativa de espécies de plantas daninhas com dois cultivares de soja

Um experimento para avaliação da competição relativa de espécies de plantas daninhas, em relação a cultivares de soja de ciclos vegetativo precoce (Embrapa-48) e médio (Embrapa-62), foi instalado em Londrina-PR, em 1998/99. Foram comparadas quatro espécies daninhas: amendoim-bravo (Euphorbia heterophylla), capim-marmelada (Brachiaria plantaginea), corda-de-viola (Ipomoea grandifolia) e fedegoso (Senna obtusifolia), ajustadas às densidades de 0, 15 e 30 plantas m-2. O experimento foi conduzido em blocos casualizados, com parcelas subdivididas, fatorial 4x3x2 e quatro repetições. A emergência da soja e a das plantas daninhas foram quase simultâneas. Ambos os cultivares responderam de modo similar à competição. A produtividade do cv. Embrapa-48 foi de 2.819 kg ha-1, e a do cv. Embrapa-62, de 2.565 kg ha-1, na ausência de plantas daninhas. As intensidades relativas médias de competição foram: B. plantaginea (0,35) < I. grandifolia (0,59) < E. heterophylla (0,61) < S. obtusifolia (1,00). As estimativas de redução de produtividade de soja, feitas por meio de equações de regressão linear/cultivar, indicaram os seguintes coeficientes de redução (%) por unidade de planta daninha m-2: B. plantaginea (Y E48 = -1,47; Y E62 = -1,58); I. grandifolia (Y E48 = -2,51; Y E62 = -2,67), E. heterophylla (Y E48= -2,47; Y E62 = -2,83); e S. obtusifolia (Y E48 = 4,52; Y E62 = -4,21). Equações de ajuste de reduções de produção de soja às infestações de plantas daninhas são discutidas.

21-Hydroxylase deficiency in Brazil

We determined the frequency of large rearrangements and point mutations in 130 Brazilian patients with 21-hydroxylase deficiency and correlated genotype with phenotype. The frequency of CYP21 deletions was lower (4.4%) than in most of the previous series described, whereas the frequency of large gene conversions was similar to the frequency reported in the literature (6.6%). The most frequent point mutations were I2 splice (41.8% in salt wasting - SW), I172N (32.6% in simple virilizing - SV) and V281L (40.2% in the late onset form - LO). The frequency of the nine most common point mutations was similar to that reported for other countries. The 93 fully genotyped patients were classified into 3 mutation groups based on the degree of enzymatic activity (A<2%, B @ 2%, C>20%). In group A, 62% of cases presented the SW form; in group B, 96% the SV form, and in group C, 88% the LO form. We diagnosed 80% of the affected alleles after screening for large rearrangements and 15 point mutations. To diagnose these remaining alleles we sequenced the CYP21 gene of one patient with the SV form and identified a heterozygous G->A transition in codon 424. This mutation leads to a substitution of glycine by serine in a conserved region and was also found in a compound heterozygous state in 4 other patients. The mutation G424S presented a linkage disequilibrium with CYP21P and C4A gene deletions and HLA DR17, suggesting a probable founder effect. Search for the G424S mutation in other populations will reveal if it is restricted to the Brazilian patients or if it has a wider ethnic distribution.