Comparison of Some Molecular-genetic Techniques for Identification of Leishmania Circulating in Natural Foci of Zoonotic Cutaneous Leishmaniasis in the Central Asia Region

Different molecular-genetic methods were used to identify a cohort of Leishmania strains from natural foci of zoonotic cutaneous leishmaniasis located in Central Asia, on the former USSR territory. The results obtained using isoenzymes, PCR, restriction fragment length polymorphisms of kDNA and molecular hybridization techniques are discussed in terms of their applicability, discrimination power and feasibility for answering questions related to molecular epidemiological research and for detecting mixed Leishmania infections

Culex gnomatos a New Species of the Spissipes Section of Culex (Melanoconion) (Diptera: Culicidae) from the Amazon Region

The adult male and female of Culex (Melanoconion) gnomatos n. sp. are described and compared with Cx. vomerifer Komp, Cx. portesi Senevet & Abonnenc and Cx. sacchettae Sirivanakarn & Jakob. Available data about distribution and bionomics are presented

Chagas Disease in Ecuador: Evidence for Disease Transmission in an Indigenous Population in the Amazon Region

Two well-defined synthetic peptides TcD and PEP2 were used in a sero-epidemiological study for the detection of Trypanosoma cruzi infections in an indigenous group in the Amazon region of Ecuador. Of the 18 communities studied along the Río Napo, province of Napo, 15 (83.3%) were found to be positive for T. cruzi infection. Of the 1,011 individuals examined 61 (6.03%) resulted positive. A prevalence of infection of 4.8% was found in children aged 1-5 years. The prevalence of infection increased with age, with adults 50 years or older showing a maximum prevalence of 18.8%. Autochthonous transmission of T. cruzi is present among this isolated indigenous population

Genetic Variability and Microdistribution of Triatoma infestans Genotypes and Trypanosoma cruzi Clones in Arequipa Region (Peru)

The genetic variability of Triatoma infestans and Trypanosoma cruzi populations was studied by isoenzyme analysis in two distinct areas of Arequipa province (Peru); one, Santa Rita de Siguas, being an endemic area for Chagas' disease, the second, Arequipa, recently infected. Analysis of T. infestans genetic variability indicates, (i) temporal stability of genotypes found in Santa Rita de Siguas, (ii) high genetic differences between Arequipa and Santa Rita de Siguas populations suggesting minor contact between them, (iii) multiple origin of the T. infestans population in Arequipa, and (iv) poor dispersal capacity of T. infestans: the panmictic unit could be reduce to a house. Parasite isoenzyme analysis was performed in 29 Peruvian stocks of T. cruzi, mainly isolated from bugs taken in a single locality, Santa Rita de Siguas. The results show, (i) a high genetic polymorphism, (ii) nine different multilocus genotypes were detected and clustered in two different clades, (iii) most of the parasite isolates pertained to one of the clade and were genetically similar to those analyzed 12 years before. This sample allowed the study of the mating system of T. cruzi in strict sympatic conditions and gave more strength to the hypothesis of the clonal structure of T. cruzi populations

Effects of Bayluscide WP 70® on the Survival and Water-leaving Behaviour of Biomphalaria straminea, Snail Host of Schistosomiasis in Northeast Brazil

The toxic and behavioural effects of niclosamide (Bayluscide WP 70®) on Biomphalaria straminea from a highly endemic area of schistosomiasis in northeastern Brazil were investigated through laboratory bioassays. The LD50 and LD90 were 0.114 mg/l and 0.212 mg/l, respectively. Water-leaving behaviour occurred among 14% to 30% of the snails in the presence of sublethal doses of niclosamide and among 16% of the controls. It was concluded that both the relatively low susceptibility to niclosamide and water-leaving behaviour of local B. straminea may be responsible for the recolonization of transmission foci after mollusciciding. It was suggested that recently improved measures of snail control, such as controlled-release formulations of niclosamide and plant molluscicides should be considered in areas where snail control is recommended

Two monoclonal rat antibodies with specificity for the beta-chain variable region V beta 6 of the murine T-cell receptor.

Two rat monoclonal antibodies (mAbs), 44-22-1 and 46-6B5, which recognize an alloreactive cytotoxic clone, 3F9, have been further tested on a panel of T hybridomas and cytotoxic T-cell clones for binding and functional activities. The mAbs recognized only those cells sharing the expression of the T-cell receptor beta-chain variable region gene V beta 6 with 3F9. All V beta 6+ cells were activated by these mAbs under cross-linking conditions and their antigen-specific activation was blocked by soluble mAb. Furthermore, depletion of 46-6B5+ normal lymph node T cells eliminated all cells expressing the epitope recognized by 44-22-1 and V beta 6 mRNA.

Enhancement of autophagic flux after neonatal cerebral hypoxia-ischemia and its region-specific relationship to apoptotic mechanisms.

The multiplicity of cell death mechanisms induced by neonatal hypoxia-ischemia makes neuroprotective treatment against neonatal asphyxia more difficult to achieve. Whereas the roles of apoptosis and necrosis in such conditions have been studied intensively, the implication of autophagic cell death has only recently been considered. Here, we used the most clinically relevant rodent model of perinatal asphyxia to investigate the involvement of autophagy in hypoxic-ischemic brain injury. Seven-day-old rats underwent permanent ligation of the right common carotid artery, followed by 2 hours of hypoxia. This condition not only increased autophagosomal abundance (increase in microtubule-associated protein 1 light chain 3-11 level and punctuate labeling) but also lysosomal activities (cathepsin D, acid phosphatase, and beta-N-acetylhexosaminidase) in cortical and hippocampal CA3-damaged neurons at 6 and 24 hours, demonstrating an increase in the autophagic flux. In the cortex, this enhanced autophagy may be related to apoptosis since some neurons presenting a high level of autophagy also expressed apoptotic features, including cleaved caspase-3. On the other hand, enhanced autophagy in CA3 was associated with a more purely autophagic cell death phenotype. In striking contrast to CA3 neurons, those in CA1 presented only a minimal increase in autophagy but strong apoptotic characteristics. These results suggest a role of enhanced autophagy in delayed neuronal death after severe hypoxia-ischemia that is differentially linked to apoptosis according to the cerebral region.

Power laws and scaling of rain events and dry spells in the Catalonia region

We analyze the statistics of rain-event sizes, rain-event durations, and dry-spell durations in a network of 20 rain gauges scattered in an area situated close to the NW Mediterranean coast. Power-law distributions emerge clearly for the dryspell durations, with an exponent around 1.50 ± 0.05, although for event sizes and durations the power-law ranges are rather limited, in some cases. Deviations from power-law behavior are attributed to finite-size effects. A scaling analysis helps to elucidate the situation, providing support for the existence of scale invariance in these distributions. It is remarkable that rain data of not very high resolution yield findings in agreement with self-organized critical phenomena.

Catabolic ornithine carbamoyltransferase of Pseudomonas aeruginosa. Importance of the N-terminal region for dodecameric structure and homotropic carbamoylphosphate cooperativity.

Pseudomonas aeruginosa has an anabolic (ArgF) and a catabolic (ArcB) ornithine carbamoyltransferase (OTCase). Despite extensive sequence similarities, these enzymes function unidirectionally in vivo. In the dodecameric catabolic OTCase, homotropic cooperativity for carbamoylphosphate strongly depresses the anabolic reaction; the residue Glu1O5 and the C-terminus are known to be essential for this cooperativity. When Glu1O5 and nine C-terminal amino acids of the catabolic OTCase were introduced, by in vitro genetic manipulation, into the closely related, trimeric, anabolic (ArgF) OTCase of Escherichia coli, the enzyme displayed Michaelis-Menten kinetics and no cooperativity was observed. This indicates that additional amino acid residues are required to produce homotropic cooperativity and a dodecameric assembly. To localize these residues, we constructed several hybrid enzymes by fusing, in vivo or in vitro, the E. coli argF gene to the P. aeruginosa arcB gene. A hybrid enzyme consisting of 101 N-terminal ArgF amino acids fused to 233 C-terminal ArcB residues and the reciprocal ArcB-ArgF hybrid were both trimers with little or no cooperativity. Replacing the seven N-terminal residues of the ArcB enzyme by the corresponding six residues of E. coli ArgF enzyme produced a dodecameric enzyme which showed a reduced affinity for carbamoylphosphate and an increase in homotropic cooperativity. Thus, the N-terminal amino acids of catabolic OTCase are important for interaction with carbamoylphosphate, but do not alone determine dodecameric assembly. Hybrid enzymes consisting of either 26 or 42 N-terminal ArgF amino acids and the corresponding C-terminal ArcB residues were both trimeric, yet they retained some homotropic cooperativity. Within the N-terminal ArcB region, a replacement of motif 28-33 by the corresponding ArgF segment destabilized the dodecameric structure and the enzyme existed in trimeric and dodecameric states, indicating that this region is important for dodecameric assembly. These findings were interpreted in the light of the three-dimensional structure of catabolic OTCase, which allows predictions about trimer-trimer interactions. Dodecameric assembly appears to require at least three regions: the N- and C-termini (which are close to each other in a monomer), residues 28-33 and residues 147-154. Dodecameric structure correlates with high carbamoylphosphate cooperativity and thermal stability, but some trimeric hybrid enzymes retain cooperativity, and the dodecameric Glu1O5-->Ala mutant gives hyperbolic carbamoylphosphate saturation, indicating that dodecameric structure is neither necessary nor sufficient to ensure cooperativity.

Potential Vectors of Dirofilaria immitis (Leidy, 1856) in Itacoatiara, Oceanic Region of Niterói Municipality, State of Rio de Janeiro, Brazil

Dirofilaria immitis is a widespread mosquito-borne parasite that causes dirofilariasis, a commonly diagnosed disease of dogs that is rarely reported in cats and humans. A mosquito survey was conducted in Itacoatiara in the State of Rio de Janeiro, from March 1995 to February 1996, using canine, feline and human baits. A total of 3,667 mosquitoes were dissected for D. immitis larvae, representing 19 species and 10 genera. From those, Ae. scapularis, Ae taeniorhynchus, Cx. quinquefasciatus, Cx. declarator, Cx. saltanensis and Wy. bourrouli were found infected with D. immitis parasites, and among those, only the first three harbored infective larvae. The majority of larvae were found in the Malpighian tubules (889/936), and larval melanization was observed in the two Aedes species. In descending order, the best vectors were Ae. scapularis, Ae. taeniorhynchus, and Cx. quinquefasciatus which alternate seasonally in importance. Cx. quinquefasciatus is suggested to be a vector to cats. The potential transmission of D. immitis parasites by these three vectors to man is discussed.

Humoral Immune Response to the Anti-malaria Vaccine SPf66 in the Colombian Atrato River Region

The immunogenicity of anti-malaria synthetic vaccine SPf66 was tested in a region of the Colombian middle Atrato river. The specific serum antibodies against SPf66 were quantified in vaccinees and placebo injected controls for a two-years period post-immunization. The frequency of individuals showing seroconversion of anti-SPf66 antibodies three months after completion of the immunization schedule was higher in vaccinees than in controls (52.7% and 25.5%, respectively, p<0.01). However, an over than four-fold increase of the specific anti-SPf66 antibody titers was observed only in 1.4% of vaccinees and 0.2% of the controls (p<0.01). The anti-SPf66 antibody titers augmented in vaccinees from first dose application to three months after the third dose, continuously decreasing thereafter to reach below baseline values two years after completion of the immunization schedule. The results show that SPf66 has very low immunogenicity and induces a short term humoral immune response (six months).

Lymphatic Filariasis in Brazilian Urban Area (Maceió, Alagoas)

A cross-sectional survey conducted among evening students was used to determine the prevalence of Wuchereria bancrofti infection in Maceió, capital of the State of Alagoas, northeast Brazil. A single thick-blood smear was used, being collected between 10 p.m. and 12 a.m. From a total of 29,551 students enrolled at evening elementary schools in the 33 city sectors, 16,569 (56.4%) were random selected for inclusion in the study. From those, 10,857 (65.5%) were interviewed and examined and 73 (0.7%) were found to have microfilaraemia. Autochthonous W. bancrofti carriers live in 10 of the 33 city sectors, suggesting a focal distribution. Moreover, 84% of infections were diagnosed among 29% of all students examined, inhabiting three contiguous sectors at the city central area, presenting infection rates up to 5.3%. Students living in city sectors with prevalence of microfilariae carriers greater than 1% were found to have a higher risk for infection when compared to students from the rest of the town [Relative Odds (RO) 12.8, 95% CI 6.7 - 25.1]. Eleven positive individuals from non endemic areas were living in Maceió for more than 10 years; time of residence in the area was a major risk factor for infection among students not born in the region (p<0.01). Regarding sex, male students presented a higher proportion of positive (RO 1.7, 95% CI 1.1 - 2.9).

Schistosomiasis control based on repeated chemotherapy in a rural village of the sugar-cane zone in Northeast Brazil

A schedule of repeated chemotherapy with oxamniquine, consisting of biannual treatment of school-aged (7-13 years) children and annual treatment of all other age groups, was used in a representative rural village from a highly endemic area of schistosomiasis in Pernambuco. Significant reductions in infection were obtained only after two cycles of treatment, as the overall prevalence decreased from 72.6% to 41.7% and the geometric mean egg counts per gram of faeces among positives fell from 188.4 to 76. In a school-aged cohort (n=29) three treatments at six-month intervals were necessary to significantly reduce the proportion of positives (from 75.9% to 51.7%). In a cohort of children under 7 years of age (n=20) the proportion of positives actually increased (from 30% to 45%) despite two annual treatments. Water contact was intense and host snail density was relatively high. As there is no short-term perspective of improved sanitation, auxiliary measures such as focal mollusciciding are needed for an adequate control of schistosomiasis in this and alike areas.