Allele frequencies in Azuay Population in Ecuador.

One hundred and eighty-two samples of unrelated people who requested the paternity test at the Molecular Biology and Genetics Laboratory of the Catholic University of Cuenca-Ecuador in the province of Azuay were studied, except for the D1S1656 (180 samples) and SE33 (89 samples) markers. The STRs D22S1045, D3S1358, VWA, D16S539, D2S1338, D8S1179, D21S11, D18S51, D19S433, TH01, FGA, D1S1656, D12S391, D10S1248, D2S441, and SE33 were typed from blood samples, amplifying the DNA by polymerase chain reactions and electrophoresis. The allele frequencies were estimated by simple counting and the impartial heterozygosity was also calculated. The Hardy-Weinberg equilibrium theory was studied. In the results obtained with the analyzed markers, the largest number of alleles can be observed in the markers with the highest polymorphic information content (PIC): D21S11, D16S539, D2S1338, D19S433, D18S51, FGA, D1S1656, and D12S391. In addition, SE33 was analyzed in certain samples, showing as result a high PIC, in fact, the highest one because of its great polymorphisc characteristic. Likewise, these markers are the ones providing the highest probability of discrimination and the lowest probability of coincidence.

Lateral thoracic expansion in a preterm baby with asphyxiating thoracic dystrophy.

Lateral thoracic expansion is a surgical technique which consists of increasing the diameter of the thoracic rib cage by the division of ribs and underlying tissue in a staggered fashion. To our knowledge, this procedure has not yet been described in preterm babies. We report a case of a 32-week preterm baby who was initially treated sequentially with resection of the costal cartilages and sternal spreading with the interposition of cartilage grafts, followed by left and right lateral thoracic expansion. The patient survived for 4 months after birth, showing that this procedure can be performed at any age after delivery.

Avaliação preliminar de óleos essenciais de plantas como repelentes para Aedes albopictus (Skuse, 1894) (Diptera: Culicidae) / Preliminary evaluation of plant essential oils as repellents against Aedes albopictus (Skuse, 1894) (Diptera: Culicidae)

Óleos essenciais de plantas têm sido formulados de forma caseira ou colocados no mercado de forma artesanal para repelir mosquitos hematófagos. O presente trabalho avaliou o possível efeito repelente de alguns excipientes normalmente usados em formulações comerciais e de óleos essenciais de nove plantas, sobre fêmeas de Aedes albopictus. Os testes foram realizados em caixas contendo mosquitos, oferecendo-se uma área definida de dedos da mão tratados ou não. Exceto pelo creme Lanatte e os géis de carbopol, as substâncias excipientes avaliadas mostraram ação repelente. Apenas os óleos provenientes de citronela (5 e 10 por cento) resultaram em repelência expressiva, com índices médiosde proteção acimade 98 por cento. Os resultados indicam que a metodologia empregada é adequada para a avaliação preliminar de produtos com potencial para a repelência de mosquitos.

Plant essential oils have been domestically formulated or marketed as handcraft products to repel hematophagous mosquitoes. This work evaluated the possible repellent effect of both excipients normally used in commercial formulations and essential oils from nine plants against Aedes albopictus females. The assays were carried out in boxes containing mosquitoes, where a defined area of treated or non-treated fingers was offered. Except for Lanatte cream and carbopol gels, the evaluated excipients had repellent action. Only the oils from citronella (5 and 10 percent) presented expressive repellent action, with mean protection indexes above 98 percent. The results indicate that the used methodology is appropriate for the preliminary evaluation of products with potential to repel mosquitoes.

Chemical control of Loxosceles intermedia (Araneae: Sicariidae) with pyrethroids: field and laboratory evaluation.

Bites from the recluse or brown spiders (genus Loxosceles) can cause necrotic lesions and systemic effects in humans throughout the world. In the state of Paraná, Brazil, loxoscelism is considered a serious public health problem, and Loxosceles intermedia Mello-Leitão (Araneae: Sicariidae) is associated with the majority of reported accidents. In the present research we evaluated the susceptibility of L. intermedia to pyrethroid insecticides currently used for the control of spiders in both field and laboratory conditions. In laboratory tests, the most active pesticides in descending order were microencapsulated lambda-cyhalothrin (LC50 = 0.023 mg/kg), nonmicroencapsulated lambda-cyhalothrin (LC50 = 0.047 mg/kg), deltamethrin (LC50 = 0.26 mg/kg), and cypermethrin (LC50 = 1.38 mg/kg). Cockroaches, Phoetalia circumvagans (Burmeister) (n = 30), killed with microencapsulated lambdacyalothrin, were offered to the spiders. L. intermedia fed on 63.3% of the dead cockroaches during the first 6 h of experiment; none of the spiders died during the subsequent 15 d. Microencapsulated lambdacyalothrin was chosen for application in two contiguous houses. The mean volume applied was 22.8 mg (AI)/m2. Dead spiders were found during all the inspections up to 60 d after the initial application. In total, 297 dead spiders were collected; 65.7% in the attic shared by the two homes, 10.8% inside the house that had most cracks and crevices sealed and 23.6% in the control house. The use of lambda-cyhalothrin-based products for L. intermedia control is discussed.

The potential protective effect of low potassium dextran against lipid peroxidation in a rat lung transplantation model.

The overproduction of reactive oxygen species plays an important role in the cascade of events during lung ischemia-reperfusion leading to graft failure. An evaluation of the peripheral markers of oxidative stress and antioxidant enzyme activities was carried out after reperfusion in a rat lung transplant model. The decrease in lipid peroxidation immediately after transplantation ( P < 0.05) may suggest an adaptative response and/or a protective effect of low potassium dextran against lipid peroxidation through natural scavenging mechanisms.

Thymic alterations in Plasmodium berghei-infected mice.

The primary function of the thymus is to develop immature T-cells into cells that further in the periphery will be able to carry out immune functions. The Literature has shown that thymus can be a target for many pathogens and severe structural alterations take place in this organ during infectious diseases. Here, we investigated if thymus is also a target organ during experimental malaria infection by analyzing the presence of parasites inside the organ and histological alterations in thymuses from Plasmodium berghei NK65-infected BALB/c. After 14 days of infection, parasites were found inside the thymus that presented a profound atrophy with total loss of its architecture. We propose that the presence of parasites in the thymus induces histological modifications that alter the microenvironment, impairing by consequence the successful T cell development. Additional studies are currently being developed in our laboratory to verify if such thymic alterations can influence the systemic immune response to the parasite.

Neisseria meningitidis PorA variable regions: rapid detection of P1.7 and P1.19 variants by PCR.

AIM: Rapid characterization of variable region (VR)1 variants of the porA gene among invasive strains is crucial for outbreak management and epidemiology studies. Recent sequence analysis studies in Brazil showed that the VR1 P1.7 and P1.19 variants are highly prevalent, accounting for 68%, of the total number of VR1 variants characterized. The aim of this work is to develop a rapid polymerase chain reaction (PCR)-based method for genosubtyping Neisseria meningitidis by detection of porA variable regions P1.7 and P1.19. METHODS AND RESULTS: PCR primers for the detection of porA VR1 P1.7 and P1.19 were designed and tested using 198 clinical N. meningitidis isolates that had been previously evaluated by porA sequencing. All 50 strains with VR1 P1.7 and all 65 strains with VR1 P1.19 were positively identified by the respective VR-specific PCR and no false-positive reactions occurred. CONCLUSIONS: VR-specific PCR amplification accurately identified VR P1.7 and P1.19 strains. SIGNIFICANCE AND IMPACT OF THE STUDY: To overcome the disadvantages of serosubtyping and sequencing for typing the porA VR1 segment of N. meningitidis, we developed a PCR-based method to rapidly and accurately detect VR1 P1.7 and P1.19 variants. This approach is highly specific and sensitive; moreover it may allow for genotype determination of culture-negative samples.

DNA microarray analysis of gene expression in alveolar epithelial cells in response to TNFalpha, LPS, and cyclic stretch.

Recent evidence suggests that alveolar epithelial cells (AECs) may contribute to the development, propagation, and resolution of acute lung injury (ALI)/acute respiratory distress syndrome (ARDS). Proinflammatory cytokines, pathogen products, and injurious mechanical ventilation are important contributors of excessive inflammatory responses in the lung. In the present study, we used cDNA microarrays to define the gene expression patterns of A549 cells (an AEC line) in the early stages of three models of pulmonary parenchymal cell activation: cells treated with tumor necrosis factor-alpha (TNFalpha) (20 ng/ml), lipopolysaccharide (LPS, 1 microg/ml), or cyclic stretch (20% elongation) for either 1 h or 4 h. Differential gene expression profiles were determined by gene array analysis. TNFalpha induced an inflammatory response pattern, including induction of genes for chemokines, inflammatory mediators, and cell surface membrane proteins. TNFalpha also increased genes related to pro- and anti-apoptotic proteins, signal transduction proteins, and transcriptional factors. TNFalpha further induced a group of genes that may form a negative feedback loop to silence the NFkappaB pathway. Stimulation of AECs with mechanical stretch changed cell morphology and activated Src protein tyrosine kinase. The combination of TNFalpha plus stretch enhanced or attenuated expression of multiple genes. LPS decreased microfilament polymerization but had less impact on NFkappaB translocation and gene expression. Results from this study indicate that AECs can tailor their response to different stimuli or/and combination of stimuli and subsequently play an important role in acute inflammatory responses in the lung.

[Larval susceptibility to chemical insecticides of two Aedes aegypti populations]. / Susceptibilidade larval de duas populações de Aedes aegypti a inseticidas químicos.

OBJECTIVE: Insect susceptibility has been one of the most important aspects to be monitored in public health programs for vector control. The purpose of the study is to assess the susceptibility to chemical insecticides of Aedes aegypti larvae in both areas under vector control and no vector control. METHODS: World Health Organization standard bioassays for diagnostic concentration and multiple concentrations were performed in mosquito larvae collected in an area under no vector control (Campinas, SP) and an area under vector chemical control (Campo Grande, MS), in Brazil. RESULTS: Potential resistance to a diagnostic concentration of temephos (DC=0.04 ppm) was registered for an Ae. aegypti larval population collected in Campinas. Multiple concentration tests confirmed the larvae resistance, with 24.5% of them surviving at the 0.0125 ppm concentration. Bioassays with the organophosphate fenitrothion (DC=0.08 ppm) and pyrethroid cypermethrin (DC=0.01 ppm) in the same population revealed their susceptibility to these agents. Bioassays carried out in an Ae. aegypti larval population collected in Campo Grande showed their susceptibility to temephos (DC=0.04 ppm) and cypermethrin (DC=0.01 ppm). LC50 and LC95 for cypermethrin (CE25), cyfluthrin (CE5), betacyfluthrin (SC1.25) and propoxur (CE20) were determined for Ae. aegypti. Using the Rockefeller standard strain values, ratios of resistance were estimated: 2.9, 2.2, 2.4 and 1.3 for LC50 and 3.5, 2.6, 3.9 and 1.3 for LC95, respectively. CONCLUSION: The findings reinforce the need for routinely monitoring pesticide efficacy as a very important step in vector control management programs.

Differential predation of the planarian Dugesia tigrina on two mosquito species under laboratory conditions.

Two experiments were performed on the predation of the planarian Dugesia tigrina (Girard) upon 2 mosquito prey species, Aedes albopictus and Culex quinquefasciatus. Bioassays were carried out in sectioned tires with 2 liters of water. In the 1st experiment, predation was evaluated using 4, 8, and 12 mature planarians against 40 2nd-stage larvae of each mosquito species alone. In the 2nd experiment, the same 3 predator densities were used with a pool composed of 20 2nd-stage larvae of each mosquito species. In the 1st experiment, final corrected mortality of Ae. albopictus reached 89.1, 98.8, and 99.6% and final corrected mortality of Cx. quinquefasciatus reached 29.4, 48.0, and 53.0%, respectively, with 4, 8, and 12 planarians. In the 2nd experiment and when subjected to the density of 4 planarians, Ae. albopictus was more susceptible to predation, with a selectivity index of 0.87, whereas this index was 0.13 for Cx. quinquefasciatus. Predation was more intensive during the 1st 4 days of the experiments, when most larvae were in the 2nd and 3rd stages. We observed that Cx. quinquefasciatus larvae were faster than Ae. albopictus in reacting to planarian contacts, resulting in more success in escaping from the predator attacks.

Chromosomal comparisons among and within populations of Simulium (Chirostilbia) pertinax (Diptera, Simuliidae).

Chromosomal studies were carried on six larval populations of Simulium (Chirostilbia) pertinax from different locations in Brazil. Larvae were collected in the states of Paraná, Rio Grande do Sul, Rio de Janeiro and São Paulo. Polytene chromosome map comparisons within and among populations showed no differences in banding pattern, except for some limited polymorphism (secondary NOR and four band polymorphisms). There were no chromosomal variations associated with the resistance or susceptibility of the larvae to temephos. The chromosomal homosequentiality found among the six populations suggests that S. pertinax may be a monomorphic species.

[Search of natural occurrence of xiphidiocercariae (trematoda) in fresh water snails of nine countries from São Paulo State, Brazil]. / Búsqueda de xifidiocercarias (Trematoda) en moluscos de agua dulce recolectados en nueve municipios del Estado de São Paulo, Brasil.

Xiphidiocercariae, aquatic larval stages of some trematodes are considered a potential instrument for biological control of mosquitoes. In this study we evaluated its natural occurrence in Campinas region and two places in Vale do Ribeira (Registro and Miracatu), São Paulo State. Snails were obtained from fresh water collections from September 1996 to February 1999. The species collected were Lymnaea columella, Physa marmorata, Biomphalaria tenagophila, Biomphalaria sp., Drepanotrema cimex, D. lucidum and Drepanotrema sp. Fasciola hepatica, xiphidiocercariae (Haematoloechidae) and echinostomatid cercariae were detected in the lymnaeids snails from Miracatu, SP. In the same locality were found planorbids snails parasitized by furcocercariae, echinostomatid cercariae and xiphidiocercariae. The xiphidiocercariae found in the planorbids were different from those obtained from lymnaeids. One Biomphalaria sp. infected with furcocercariae was found in Louveira, SP. In the ROSA place (Campinas, SP) an individual of Biomphalaria sp. and one of L. columella were found infected by the furcocercariae and echinostomatid cercariae, respectively. In the place UNI-I, in Campinas, one L. columella was infected by furcocercariae. Double infection in snails from Miracatu was also observed.

Differential diagnosis of chest pain in an emergency department: the value of homocysteinemia as a diagnostic marker.

UNLABELLED: Only 20 to 45% of patients (pts) admitted to an emergency department (ED) with chest pain (CP) have a cardiac ischemic CP (ICP). Apart from the need for a rapid detection of ICP cases, it is important to avoid discharging patients with false negative ICP. We studied the capability of plasma homocysteine (Hcy) values to improve the differential diagnosis (DD) in patients admitted to an ED with CP. PATIENTS: 125 patients, 80 males, mean age 61 yrs (25-96 yrs), 75 (60%) without prior atherosclerotic cardiovascular disease (ACVD), consecutively admitted to the ED with CP which was not immediately clear. Definitive diagnosis: ICP-58 patients (46%); non-ischemic (non-ICP)--64 patients (54%). METHOD: Hcy was measured on admittance (fasting state not required), by means of a fluorescence polarization enzyme immunoassay (IMx, Abbott); at real conditions, the results were obtained within 2 hours (but not displayed). RESULTS: 1) Hcy (mumol/L) = 10.9 +/- 5.4 (non ICP without prior CDAV); 13.9 +/- 7.7 (ICP). 2) Hcy > or = 15.0 mumol/L in patients without prior ACVD: 5 patients (9%) with non ICP, 8 patients (38%) with ICP--p < 0.01; RR = 2.9 (95% CI = 1.2-7.1); positive value = 62%; negative predictive value = 79%. CONCLUSION: In patients without ACVD, Hcy may contribute to improve DD of equivocal CP, namely: 1) before a normal or non-diagnostic ECG and negative ischemia markers, Hcy > or = 15.0 mumol/L will imply additional cardiological investigation in the ED; 2) Hcy < 15.0 increases the liability of a non-ICP diagnosis.

Survey of cyclopids (Crustacea, Copepoda) in Brazil and preliminary screening of their potential as dengue vector predators.

INTRODUCTION: Cyclopid copepods are known to be good mosquito controllers, specially as regards the larvae of the dengue vectors Aedes aegypti and Ae. albopictus. MATERIAL AND METHOD: The objective of the study was to survey the local copepod fauna and search for new strains of M. longisetus var. longisetus, comparing the potential of the samples found with the current strain ML-01 against Ae. albopictus larvae, under laboratory conditions. Eleven bodies of water in Campinas, SP, Brazil, were screened for copepods by collecting 1.5 l of water from each of then. The predatory potential of adults copepods was evaluated over 24 h, in the laboratory, for groups of 5 individuals preying upon 30 first instar Ae. albopictus larvae. RESULTS AND CONCLUSION: The following cyclopid species were found: Metacyclops mendocinus, Tropocyclops prasinus, Eucyclops sp, Eucyclops serrulatus, Eucyclops solitarius, Eucyclops ensifer, Macrocyclops albidus var. albidus and Mesocyclops longisetus var. longisetus. The predatory potential of these copepods ranged from nil to 97.3%. A sample collected in the field containing only M. longisetus var. longisetus showed the best control efficiency with no significant difference from a three-year old laboratory culture (ML-01) of the same species evaluated for comparison. The sample with few M. albidus var. albidus was ranked in second place showing an average 25.9% efficiency. The use of copepods in trap tires as dengue vector controllers is discussed.

[Susceptibility of populations of Simulium (Chirostilbia) pertinax Kollar, 1832 (Culicomorpha, Simuliidae) to eemephos and to Bacillus thuringiensis var. israelensis-based formulation]. / Susceptibilidade de populações de Simulium (Chirostilbia) pertinax Kollar, 1832 (Culicomorpha, Simuliidae) ao temephos e a um formulado à base de Bacillus thuringiensis var. israelensis.

The use of wooden troughs on stream beds, artificially colonized by blackfly larvae, is proposed for larvicide evaluations. Mortality was recorded 3 or 4 hours after treatment. Larval susceptibility was also evaluated utilizing the LT50 criterion. In there field assays Simulium (C.) pertinax populations from the litoral of S. Paulo and Rio de Janeiro States were shown to be resistant to temephos, even when subjected to high concentrations. Vectobac 12 AS, a Bacillus thuringiensis var. israelensis product, was shown to be more potent against late instar larvae and efficient in concentrations higher than 7,200 ITU/l (10 min). The LT50 to 3,744 ITU/l (10 min) was calculated as 70.9 min.

[Evaluation of the sensitivity of adult Culex quinquefasciatus Say to chemical insecticides]. / Avaliação da sensibilidade de adultos de Culex quinquefasciatus Say a inseticidas químicos de contato.

The sensitivity of the adult house mosquito Culex quinquefasciatus to 5 chemical insecticides was evaluated under laboratory conditions, based on the Median Lethal Time (LT50) criterion. The organophosphorous Malathion and four pyrethroids: Bifenthrin, Deltamethrin, Esfenvalerate and Alfamethrin were utilized. An easy and efficient technique was suggested for the testing of one-day-old adults, including five repetitions for each treatment. The results revealed the full adequacy of this method for routine use. Further, no resistance to the 5 chemical compounds was detected among this natural population of Cx. quinquefasciatus.

Methods for field detection of resistance to temephos in simuliids. Larval esterase level and topical application of the insecticide to adults.

Two practical field methods for indirect detection of simuliid populations resistant to temephos are proposed. The first is based on high esterase activity in resistant larvae and involves adaptations of a filter paper test in which faintly stained spots indicate susceptible populations and strongly stained ones reveal populations resistant to temephos. The second is based on the resistance to the larvicide when adults are topically exposed, and involves the use of diagnostic doses obtained by the comparison between the LD50 for susceptible and resistant populations. The relevance of such methods is discussed in order to help resistance detection in Simulium pertinax Kollar control programmes.