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Contaminant-related changes in antioxidative processes in the freshwater crustacea Daphnia magna exposed to model redox cycling contaminant were assessed. Activities of key antioxidant enzymes including catalase, superoxide dismutase, glutathione peroxidase and glutathione S-transferases and levels of lipid peroxidation measured as thiobarbituric acid-reactive substances (TBARS) and lipofucsin pigment content were determined in D. magna juveniles after being exposed to sublethal levels of menadione, paraquat, endosulfan, cadmium and copper for 48 h. Results denoted different patterns of antioxidant enzyme responses, suggesting that different toxicants may induce different antioxidant/prooxidant responses depending on their ability to produce reactive oxygen species and antioxidant enzymes to detoxify them. Low responses of antioxidant enzyme activities for menadione and endosulfan, associated with increasing levels of lipid peroxidation and enhanced levels of antioxidant enzyme activities for paraquat, seemed to prevent lipid peroxidation, whereas high levels of both antioxidant enzyme activities and lipid peroxidation were found for copper. For cadmium, low antioxidant enzyme responses coupled with negligible increases in lipid peroxidation indicated low potential for cadmium to alter the antioxidant/prooxidant status in Daphnia. Among the studied enzymes, total glutathione peroxidase, catalase and glutathione S-transferase appeared to be the most responsive biomarkers of oxidative stress.

Age-related changes in the balance between endogenous pro-oxidative and antioxidative processes in the freshwater cladoceran Daphnia magna (Crustacea) were assessed. The activities of key antioxidant enzymes including catalase, superoxide dismutase and glutathione peroxidase and levels of lipid peroxidation measured as thiobarbituric acid-reactive substances (TBARS) were determined in eight age classes, covering juvenile, young and senescent adults. Age-related changes in fatty acid composition were also measured to examine the contribution of polyunsaturated fatty acids (PUFA) in the peroxidation status of animals. Biochemical responses depicted in this study demonstrated that age-related decline in survival was accompanied by increasing oxidative stress and oxidative damage. Enhanced oxidative stress in aging D. magna was suggested by the significant increase in the formation of lipid peroxides, and a concomitant reduction of unsaturated fatty acids of 20 or more carbon atoms. Because aging was accompanied by selective loss of key antioxidant enzymes and small changes in the amount of PUFA, the breakdown of antioxidant defences might have directly contributed to oxidative stress, membrane lipid peroxide and a decline of survival. Indeed, the results reported here, indicate that age-related increases of lipid peroxides were at least partially due to the functional imbalance of enzymatic antioxidant defences.

Background and Purpose: Transcranial Doppler (TCD) can detect intracranial stenoses and occlusions that can help in the diagnosis and management of ischemic stroke. The accuracy parameters for lesions located in the terminal internal carotid artery (TICA) are less known, unlike other basal cerebral vessels., Patients and Methods: The authors studied consecutive patients referred for TCD who underwent contrast angiography or magnetic resonance angiography. They calculated the sensitivity, specificity, positive and negative predictive values (PPV and NPV), and likelihood ratios., Results: Forty-three patients had TCD and angiography: mean age was 57 +/- 20 years, and 65% were men. Twenty-two patients were diagnosed with TICA stenosis or occlusion on TCD underwent angiography. Four patients had abnormal TCD findings that were not confirmed by angiography. Two of 21 patients with normal TCD showed moderate (< 50%) stenosis of the TICA and cavernous segment of the internal carotid artery at angiography. Accuracy parameters for TCD were as follows: sensitivity = 90% (confidence interval [CI], 63%-96%), specificity = 83% (CI, 61%-94%), PPV = 82%, NPV = 86%, positive likelihood ratio = 5, and negative likelihood ratio = 0.17., Conclusions: TCD is a sensitive screening tool for the lesions in the TICA. Specificity is likely affected by a wide spectrum of the stenosis severity shown at angiography and time lags between the studies.

Members of the New World mosquito subgenus Melanoconion of the genus Culex are important vectors of many alphaviruses including eastern and Venezuelan equine encephalitis viruses (VEEV). We investigated the phylogenetic relationships among nine putative species of the Vomerifer and Pedroi Groups of the Neotropical Spissipes Section by sequencing the internal transcribed spacer 2 (ITS-2) region of ribosomal DNA and using phylogenetic analyses. Results demonstrated that, within the Spissipes Section, the Vomerifer and Pedroi Groups are monophyletic sister groups. The clade comprised by Culex adamesi and Culex ribeirensis showed a sister group relationship to the group consisting of Culex pedroi sequences. The monophyly of the Vomerifer Group corroborated previous suggestions that it is a natural group. However, our topology showed that there are two well-supported, divergent groups within a major clade consisting of Cx. pedroi sequences, suggesting the possibility of a cryptic Cx. pedroi-like species. This finding could have important epidemiological implications for VEEV transmission in Central and South America.

In this article we present the case of a 72 year-old woman who three years after laparoscopic cholecystectomy develops obstructive jaundice. An MRI of the liver and biliary system revealed an hiliar mass that caused dilatation of the biliary tree. The patient underwent hepatic duct resection and reconstruction via hepaticojejunostomy. The histological examination of the surgical specimen identified an intramural biliary neuroma with no evidence of malignancy.

Venezuelan equine encephalitis virus (VEEV) remains a naturally emerging disease threat as well as a highly developed biological weapon. Recently, progress has been made in understanding the complex ecological and viral genetic mechanisms that coincide in time and space to generate outbreaks. Enzootic, equine avirulent, serotype ID VEEV strains appear to alter their serotype to IAB or IC, and their vertebrate and mosquito host range, to mediate repeated VEE emergence via mutations in the E2 envelope glycoprotein that represent convergent evolution. Adaptation to equines results in highly efficient amplification, which results in human disease. Although epizootic VEEV strains are opportunistic in their use of mosquito vectors, the most widespread outbreaks appear to involve specific adaptation to Ochlerotatus taeniorhynchus, the most common vector in many coastal areas. In contrast, enzootic VEEV strains are highly specialized and appear to utilize vectors exclusively in the Spissipes section of the Culex (Melanoconion) subgenus.

Biomagnification of organochlorine compounds (pesticides and polychlorinated biphenyls) through the marine aquaculture food chain is investigated. From first-feeding, specimens of sea bass were exposed to commercial fish feed (that contained DDTs and PCBs residues) for ca. 24 months, and selected tissues (white and red muscle, liver, and visceral fat) were analyzed after 6 and 24 months of diet exposure. Data obtained showed that experimental fish tissues presented a similar contamination pattern to that of fish feed, and biomagnification processes of these compounds were proved. Additionally, commercial sea bass cultured in farms from the western Mediterranean were analyzed, their organochlorine concentrations being significantly lower than those of the 24 month old experimental fish. Thus, the exposition of human population to OCs through consumption of cultured fish would be lower than expected from experimental biomagnification studies, although red muscle presented similar OC levels in both cases, which were much higher than those of white muscle. Although levels of organochlorine compounds were found to be low, the persistence, ubiquity, and toxicity of these compounds, together with their presence in fish feed, make it necessary to monitor OC residues in the routine quality assurance programs of aquaculture activities, as this food chain is a source of these toxic compounds for human consumers. The development of sensitive analytical methodology based on GC-MS/MS has allowed for the reaching of low detection limits required to carry out the present study.

Seven amino-acid substitutions introduced into the 343 amino-acid-long sequence of Coprinus cinereus peroxidase (CiP) led to a mutant enzyme (TS-rCiP) which is more stable than the native enzyme at higher temperature, pH and hydrogen peroxide concentrations. It is therefore more suitable for industrial applications. A structure determination was conducted on a deglycosylated but still active form of TS-rCiP based on X-ray diffraction data to 2.05 A resolution measured on a crystal cooled to 100 K and refined to R = 0.202 and R(free) = 0.249. The increased stability of the TS-rCiP enzyme can be understood from the structural changes of the TS-rCiP structure revealed by a comparative analysis with other known CiP structures. One of the more significant changes caused by three of the substitutions, I49S, V53A and T121A, is the conversion of a hydrophobic pocket into a hydrophilic pocket with associated changes in the water structure and the hydrogen-bonding interactions. The E239G substitution, which gives rise to increased thermostability at high pH, creates changes in the water structure and in the orientation of a phenylalanine (Phe236) in its vicinity. The three substitutions M166F, M242 and Y242F introduced to increase the oxidative stability do not introduce any structural changes.

The structure of the peroxidase from Coprinus cinereus (CiP) has been determined in three different space groups and crystalline environments. Two of these are of the recombinant glycosylated form (rCiP), which crystallized in space groups P2(1)2(1)2(1) and C2. The third crystal form was obtained from a variant of CiP in which the glycosylation sites have been removed (rCiPON). It crystallizes in space group P2(1) with beta approximately 90 degrees; the structure was determined from room-temperature data and low-temperature data obtained from twinned crystals. Two independent molecules of CiP related by non-crystallographic symmetry are contained in the three crystal forms. The packing in the two structures of the glycosylated form of rCiP is closely related, but differs from the packing in the unglycosylated rCiPON. A database search based on small-molecule porphinato iron (III) complexes has been performed and related to observations of the spin states and coordination numbers of the iron ion. The room-temperature structures of CiP and one structure of the almost identical peroxidase from Arthromyces ramosus (ARP) have been used to identify 66 conserved water molecules and to assign a structural role to most of them.

Traditional staining and microscopic examination techniques for the detection of Mycobacterium leprae, DNA amplification by polymerase chain reaction (PCR) of a 531-bp fragment of the M. leprae specific gene encoding the 36-kDa antigen, and serodiagnosis with M. leprae specific antigens (PGL-1 and D-BSA) were compared on different clinical specimens (serum samples, slit-skin smears, biopsies and swabs) from 60 leprosy patients attending the Sanatorium of Fontilles. Patients were divided into groups; (i) 20 multibacillary patients (MB) with positive bacteriological index (BI) by conventional methods and on WHO multidrug therapy (MDT); (ii) 30 MB patients with negative BI and completed minimum 2 years treatment MDT; (iii) 10 paucibacillary (PB) patients who had completed 6 months MDT at least 8 years ago. Control groups included four non-leprosy patients for PCR methods and 40 health control patients and 10 tuberculosis patients for serological methods. In the multibacillary BI positive group, there was a good correlation between all methods. All tests were negative in the paucibacillary group, although only a few patients were tested and all had been treated many years ago. One must be cautious concerning the diagnostic potential of these techniques in this type of leprosy. We also studied different combinations of leprosy diagnosis methods to determine the potential risk in a leprosy contact individuals group. The prevalence of antibodies to M. leprae antigens in serum was measured, together with the presence of M. leprae DNA in the nose and lepromin status in a group of 43 contacts of leprosy patients (12 household and 31 occupational) to evaluate the maintenance of infection reservoirs and transmission of the disease. Only two individuals were found to form a potential high risk group.

The ecology of Venezuelan equine encephalitis (VEE) virus transmission was compared at three enzootic foci: two forest sites in the Catatumbo region of western Venezuela that have yielded small numbers of virus isolates since the 1970s, and another focus in the middle Magdalena Valley of Colombia that has consistently yielded many VEE virus isolates. Our results demonstrated dramatic differences in VEE virus isolation rates from sentinel hamsters, as well as differences in mosquito species composition and captured mammals with antibodies to VEE virus, between the Colombian and Venezuelan study sites. The higher isolation rate of enzootic VEE virus in the Colombian site was associated with a more abundant fauna of spiny rats (Proechimys spp.), known reservoir hosts of enzootic VEE virus. Mosquito collections demonstrated that the Colombian forest had a higher mosquito diversity and species evenness than either of the Venezuelan forests. The Colombian focus was especially richer in its Culex (Melanoconion) spp. fauna, a subgenus that includes all proven enzootic vectors for VEE virus. Our results suggest that the greater abundance, diversity, and stability of enzootic vector populations, combined with the greater density of rodent reservoir hosts, explains the higher levels of VEE virus circulation in the Colombian focus compared with the Venezuelan forests.

In this study, the acute toxicity of the organophosphorous pesticides dichlorvos and chlorpyrifos to two different species of Artemia (A. salina and A. parthenogenetica) was evaluated. In addition, the in vivo effect of these two pesticides on cholinesterase (ChE) activity of both A. salina and A. parthenogenetica was also determined. The characterisation of the ChE, using different substrates and specific inhibitors, and the normal range of activity in non-exposed individuals were previously investigated for both species. The results obtained indicate that the ChE of A. salina is different from that of A. parthenogenetica and that both enzymes cannot be classified neither as acetylcholinesterase nor as butyrylcholinesterase since they show intermediary characteristics between the two vertebrate forms. The range of normal ChE activity was 2.65+/-0.15 U/mg protein for A. salina, and 3.69+/-0.17 U/mg protein for A. parthenogenetica. Significant in vivo effects of both pesticides on Artemia ChE activity were found, at concentrations between 5.38 and 9.30 mg/l for dichlorvos and between 1.85 and 3.19 mg/l for chlorpyrifos. Both Artemia species are resistant to these pesticides and they are able to survive with more than 80% ChE inhibition. However, A. parthenogenetica is more resistant than A. salina, with about a 95% reduction in its ChE activity respect to the control for nauplii exposed to the median lethal concentrations (LC50), without lethal effects after 24 h of exposure.

Orotidine 5'-monophosphate decarboxylase (ODCase) catalyses the decarboxylation of orotidine 5'-monophosphate to uridine 5'-monophosphate (UMP). We have earlier determined the structure of ODCase from Escherichia coli complexed with the inhibitor 1-(5'-phospho-beta-d-ribofuranosyl)barbituric acid (BMP); here we present the 2.5 A structure of the uncomplexed apo enzyme, determined from twinned crystals. A structural analysis and comparison of the two structures of the E. coli enzyme show that binding of the inhibitor is accompanied by significant domain movements of approximately 12 degrees around a hinge that crosses the active site. Hence, the ODCase dimer, which contains two active sites, may be divided in three domains: a central domain that is fixed, and two lids which independently move 12 degrees upon binding. Corresponding analyses, presented herein, of the two Saccharomyces cerevisiae ODCase structures (with and without BMP) and the Methanobacterium thermoautotrophicum ODCase structures (with and without 6-aza UMP) show very similar, but somewhat smaller domain movements. The domain movements seem to be initiated by the phosphoryl binding to the enzyme and can explain why the binding of the phosphoryl group is essential for the catalytic function., ((c) 2002 Elsevier Science Ltd.)

The accumulation and transfer of the organophosphorus pesticide chlorpyrifos has been studied in an experimental aquatic two-level food chain using two species of the crustacean Artemia (A. franciscana and A. parthenogenetica) and the small fish Aphanius iberus. Artemia adults contaminated by exposure to the pesticide in water were used as live prey for Aphanius, the next trophic level. During the experimental bioaccumulation phase, fish were fed chlorpyrifos-contaminated Artemia pools with concentrations between 6.5 and 14.5 ng/g fresh weight for 32 days. Both concentrations accumulated in fish, and biomagnification factor (BMF) values showed a continuous decrease during the bioaccumulation phase, probably due to the physicochemical characteristics of the organophosphorus pesticide, to the biotransformation ability of fish and to the progressive adaptation of fish metabolism to toxic exposure. The first day that fish were fed uncontaminated preys, the pesticide accumulated via food was rapidly eliminated and was not detected. The effect of chlorpyrifos exposure through the food chain on stress protein (HSP70) synthesis was measured as a general biochemical response of stress in the fish (A. iberus). The levels of HSP70 were significantly higher in fish fed on contaminated Artemia than in the control fish fed on uncontaminated Artemia. Results showed that the HSP70 induction in fish could be associated to exposure of chlorpyrifos via food.

During field studies of enzootic Venezuelan equine encephalitis (VEE) viruses associated with epizootic emergence, a large number of virus isolates were made in sylvatic foci of Venezuela and Colombia. To rapidly characterize these isolates, antigenic subtypes were determined by means of immunofluorescence and by single-strand conformational polymorphism (SSCP) analysis by use of an 856-bp fragment from the P62 gene, which we used to distinguish genetic variants. Representative isolates were sequenced to assess the sensitivity of SSCP to detect genetic differences. The SSCP analysis distinguished isolates differing by as little as 1 nucleotide; overall, differences of > or = 1 nucleotide were recognized 89% of the time, and the sensitivity to distinguish strains that differed by only 1 or 4 nucleotides was 17 and 57%, respectively. Phylogenetic analyses of representative sequences showed that all recent isolates from the Catatumbo region of western Venezuela and the middle Magdalena Valley of Colombia were closely related to epizootic subtype IAB and IC strains; strains from Yaracuy and Miranda States were more distantly related. Cocirculation of the same virus genotype in both Colombian and Venezuelan foci indicated that these viruses are readily transported between enzootic regions separated by > 300 km. The SSCP analysis appears to be a simple, fast, and relatively efficient method of screening VEE virus isolates to identify meaningful genetic variants.

We studied the spatial localization of mosquitoes in sylvatic focus of Venezuelan equine encephalitis virus in western Venezuela to identify mosquito species potentially involved in the hypothesized transport of viruses out of enzootic foci. The following criteria were used to identify species with potential for virus export: (1) common in the forest and surrounding area, (2) feeding on a wide range of vertebrates; (3) long dispersal capabilities, and (4) established vectorial competence for enzootic or epizootic VEE viruses. CDC traps baited with light/CO2 were operated for four and 12-h intervals to collect mosquitoes at four stations along two forest/open area transects from September to November 1997. We collected 60,444 mosquitoes belonging to 11 genera and 34 species. The most common species were Aedes serratus (Theobald), Ae. scapularis (Rondani), Ae. fulvus (Wiedmann), Culex nigripalus Theobald, Cx, (Culex) "sp", Cx. mollis Dyar & Knab, Cx. spissipes (Theobald), Cx. pedroi Sirivanakarn and Belkin, Psorophora ferox (Humboldt), Ps. albipes (Theobald), and Ps. cingulata (F.). Very few mosquitoes were captured during the (day in the open area outside the forest, suggesting that any virus export from the forest may occur at night. The following mosquitoes seemed to be mostly restricted to the forest habitat: Ae. serratus, Ps. ferox, Ps. albipes, sabethines, Cx. spissipes, Cx. pedroi, Cx. dunni Dyar, and Ae. fulvus. The main species implicated its potential virus export were Cx. nigripalpus, Ae. scapularis, and Mansonia titillans (Walker).

The distribution of the sylvatic subtype ID Venezuelan equine encephalitis (VEE) viruses in the lowland tropical forests of western Venezuela was investigated using remote sensing and geographic information system technologies. Landsat 5 Thematic Mapper satellite imagery was used to study the reflectance patterns of VEE endemic foci and to identify other locations with similar reflectance patterns. Enzootic VEE virus variants isolated during this study are the closest genetic relatives of the epizootic viruses that emerged in western Venezuela during 1992-1993. VEE virus surveillance was conducted by exposing sentinel hamsters to mosquito bites and trapping wild vertebrates in seven forests identified and located by means of the satellite image. We isolated VEE viruses from 48 of a total of 1,363 sentinel hamsters in two of the forests on six occasions, in both dry and wet seasons. None of the 12 small vertebrates captured in 8,190 trap-nights showed signs of previous VEE virus infection. The satellite image was classified into 13 validated classes of land use/vegetation using unsupervised and supervised techniques. Data derived from the image consisted of the raw digital values of near- and mid-infrared bands 4, 5, and 7, derived Tasseled Cap indices of wetness, greenness, and brightness, and the Normalized Difference Vegetation Index. Digitized maps provided ancillary data of elevation and soil geomorphology. Image enhancement was applied using Principal Component Analysis. A digital layer of roads together with georeferenced images was used to locate the study sites. A cluster analysis using the above data revealed two main groups of dense forests separated by spectral properties, altitude, and soil geomorphology. Virus was isolated more frequently from the forest type identified on flat flood plains of main rivers rather than the forest type found on the rolling hills of the study area. The spatial analysis suggests that mosquitoes carrying the enzootic viruses would reach 82-97% of the total land area by flying only 1-3 km from forests. We hypothesize that humans within that area are at risk of severe disease caused by enzootic ID VEE viruses. By contrast, equines could actually become naturally vaccinated, thus preventing the local emergence of epizootic IC VEE virus strains and protecting humans indirectly.

Bronchomalacia in adults develops secondary to bronchial disease processes (chronic bronchitis, tuberculosis, neoplasm) or to lesions resulting from prolonged intubation, surgery, trauma, or lung transplantation. We report the case of an 84-year-old woman with no history of any of the aforementioned associations in whom we detected severe idiopathic bronchomalacia of the main left bronchus. The identified lesion caused recurrent pneumonias leading to death. We show images obtained by bronchoscopy, revealing the severity of the lesion and its functional repercussions.

From 1997-1998, we investigated the possible continuous circulation of epizootic Venezuelan equine encephalitis (VEE) virus suggested by a 1983 subtype IC interepizootic mosquito isolate made in Panaquire, Miranda State, Venezuela. The study area was originally covered by lowland tropical rainforest but has been converted into cacao plantations. Sentinel hamsters, small mammal trapping, mosquito collections, and human serosurveys were used to detect active or recent virus circulation. Six strains of subtype ID VEE virus were isolated from hamsters that displayed no apparent disease. Four other arboviruses belonging to group A (Togaviridae: Alphavirus), two Bunyamwera group (Bunyaviridae), and three Gamboa group (Bunyaviridae) arboviruses were also isolated from hamsters, as well as 8 unidentified viruses. Venezuelan equine encephalitis-specific antibodies were detected in 5 small mammal species: Proechimys guairae, Marmosa spp., and Didelphis marsupialis. Mosquito collections comprised of 38 different species, including 8 members of the subgenus Culex (Melanoconion), did not yield any virus isolates. Sera from 195 humans, either workers in the cacao plantation or nearby residents, were all negative for VEE virus antibodies. Sequences of 1,677 nucleotides from the P62 gene of 2 virus isolates indicated that they represent a subtype ID lineage that is distinct from all others characterized previously, and are unrelated to epizootic VEE emergence.

The effect of cestode parasitism on the respiration rate (MO2) of different strains of Artemia from wild populations of Spain was studied. Respiration rates (MO2) of adults from each strain were not affected by the presence of cysticercoids of Flamingolepis liguloides or Hymenolepis stellorae (Cestode, Hymenolepididae). This finding could be related to the absence of reproductive activity (parasite castration) in parasitized females.

The subgeneric relationships among 8 generic and infrageneric taxa of the Culicini tribe were examined by cladistic analysis based on 30 larval mouthpart characters (maxillae and mandibles) of 7 subgenera of Culex L. and 1 of Deinocerites Theobald. We analyzed 18 ingroup species as well as Deinocerites melanophylum Dyar and Knab as a sister group, and Aedes taeniorhynchus Wiedemann as an outgroup. A parsimony analysis using the Nona program resulted in 2 trees each of 109 steps (consistency and retention indices = 0.88). The topologies obtained were similar to the current classification of the tribe, based on nonexplicit methods mainly including adult characters, with 2 exceptions. In the present work, the monophyly of the tribe Culicini (Culex + Deinocerites) was supported by 4 synapomorphies. The subgenus Lutzia Theobald formed the most basal clade in the tribe Culicini and the placement of Phenacomyia Harbach and Peyton as a subgenus was validated by its location as a sister group of the subgenus Culex and other subgenera. The subgenus Carrollia Lutz was the most robust taxon, supported by 5 synapomorphies, and was congruent with the infragroups of the current classification. The relationships among Deinocerites, Anoedioporpa Dyar, Microculex Theobald, and Melanoconion Theobald were unresolved, but were placed in the most internal clade of the tribe. The 1st exception to the accepted classification was the poorly resolved boundary between Anoedioporpa and Microculex The 2nd was the strong support (with 11 synapomorphies) for the inclusion of Deinocerites as a subgenus of Culex in the Culicini, which is proposed here.

Recent studies have indicated that epizootic Venezuelan equine encephalitis (VEE) viruses can evolve from enzootic, subtype ID strains that circulate continuously in lowland tropical forests (A. M. Powers, M. S. Oberste, A. C. Brault, R. Rico-Hesse, S. M. Schmura, J. F. Smith, W. Kang, W. P. Sweeney, and S. C. Weaver, J. Virol. 71:6697-6705, 1997). To identify mutations associated with the phenotypic changes leading to epizootics, we sequenced the entire genomes of two subtype IC epizootic VEE virus strains isolated during a 1992-1993 Venezuelan outbreak and four sympatric, subtype ID enzootic strains closely related to the predicted epizootic progenitor. Analysis by maximum-parsimony phylogenetic methods revealed 25 nucleotide differences which were predicted to have accompanied the 1992 epizootic emergence; 7 of these encoded amino acid changes in the nsP1, nsP3, capsid, and E2 envelope glycoprotein, and 2 were mutations in the 3' untranslated genome region. Comparisons with the genomic sequences of IAB and other IC epizootic VEE virus strains revealed that only one of the seven amino acid changes associated with the 1992 emergence, a threonine-to-methionine change at position 360 of the nsP3 protein, accompanied another VEE virus emergence event. Two changes in the E2 envelope glycoprotein region believed to include the major antigenic determinants, both involving replacement of uncharged residues with arginine, are also candidates for epizootic determinants.

Background: In this report we study tuberculosis transmission in HIV infected patients using molecular epidemiological methods., Patients and Methods: We have studied 60 M. tuberculosis isolates from 30 HIV infected cases, and their clinical-epidemiological data. Susceptibility to tuberculostatic agents and electrophoretic patterns using RFLPs (restriction fragment length polymorphisms) method were evaluated. Dice's coefficient was used for the similarity analysis., Results: Over 73% studied patients were included in clusters using RFLPs analysis. This data show that nearly 60% of the tuberculosis cases in our area have a recent transmission. Forty per cent of these cases were included in the main cluster. The frequency of tuberculostatic-resistant strains in HIV infected patients was similar to the that of observed in other patients. We did not find correlation between RFLPs clusters and clinical-epidemiological data., Conclusions: Tuberculosis transmission in HIV-positive patients using RFLPs as molecular marker shows that 60% of the cases are caused by recently acquired strains. We did not find multi-drug resistant strains in our isolates. However due to the high transmissibility of these circulating clones, control disease measures in this group of risk population are required.

The optimization of an arbitrarily primed PCR method for typing 96 methicillin-resistant Staphylococcus aureus (MRSA) isolates was compared with pulsed-field gel electrophoresis. Identical results in the differentiation of MRSA clones and identification of the main cluster that included 82 strains (88% of patients) were obtained by both techniques.

Objectives: To describe the personality traits and value systems of the population of residents in anesthesiology of a province in Andalusia and to analyze what the ideal profile of an anesthesiologist in training would be., Subjects and Method: Forty-four residents in anesthesiology were invited to participate, of whom 22 accepted. The participants voluntarily completed anonymous written questionnaires with 232 items: the sixteen personality factors questionnaire, form A (R.B. Cattell), and the survey of interpersonal values (L.V. Gordon)., Results: The residents scored high on primary factors of intelligence (B), self-sufficiency (Q2) and conservatism (Q1). High scores were also recorded on the secondary factors of anxiety (QI) and socialization (QIII); low scores were seen for independence (QIV). Four theoretical personality models (alpha, beta, gamma and delta) were established based on the data. Four residents were alphas (18%), 10 were betas (45%), 6 were gammas (27%) and 2 were deltas (10%)., Conclusions: An ideal personal profile for residents in anesthesiology by American authors has been available since the 1960s. An ideal resident should be independent, calm, aware, stable, secure, self-disciplined and alert; moreover, he or she should enjoy team work. The residents with alpha personalities were ideal. At present, the training of residents and the selection criteria for access to specialized training based on a personality test are scarcely valued in our discipline.

Objective: To study changes over time in body temperature related to insufflation of CO2., Patients and Method: Fifty patients were randomly assigned to 2 groups of 25 to undergo cholecystectomy by either laparoscopy or laparotomy. Total intravenous anesthesia with propofol, pancuronium and fentanyl was used in both groups. Ventilation was maintained at 0.5 FiO2. Central temperature was continuously measured by a distal esophageal thermometer and results were recorded every 10 minutes in both groups. All operations lasted approximately 80 min., Results: We found that temperature gradually decreased over time in both groups. In the laparotomy group the decrease reached 0.20 degree C (SD 0.03) at 80 min. During laparoscopy the temperature decrease was 0.43 degree C (SD 0.04) for the same time period. The differences were statistically significant. We observed no pathophysiologic repercussions associated with these results., Conclusions: Laparoscopic surgery, even when the abdominal cavity is not exposed to room air, induces a loss of temperature that is greater than that of laparotomy, because of insufflation of CO2 at 4 degrees C. The decrease was 0.4 degree C for every 50 l of CO2 insufflated during the study.

The possibility that endogenous serotonin (5-hydroxytryptamine, 5-HT) participates in alteration of the blood-brain barrier (BBB) following short-term forced swimming (FS) exercise was examined in a rat model. Subjection of conscious young (age 8-9 weeks, 80-90 g) animals to continuous FS (at a water temperature of 30 +/- 1 degrees C) for 30 min, increased the permeability of the BBB to Evans blue albumin (EBA) and 131I-sodium in six and nine brain regions, respectively. The EBA staining was noted in posterior cingulate cortex, parietal, occipital cortices, cerebellar vermis, medial lateral cerebellar cortices and dorsal surface of hippocampus. In addition to these brain regions, the BBB permeability to 131I-sodium was further extended to caudate nucleus, thalamus and hypothalamus. This effect of FS on the BBB permeability was absent in adult (age 24-30 weeks, 300-400 g) animals. Measurement of 5-HT showed a profound increase of plasma and brain in young rats by 180% and 250%, respectively, from the control group. Adult animals showed only a minor increase in brain and plasma 5-HT levels. In young animals, pretreatment with p-CPA (a 5-HT synthesis inhibitor) and indomethacin (a prostaglandin synthesis inhibitor) prevented the FS induced increase in BBB permeability and 5-HT levels. Destruction of serotonergic neurons with 5,7-dihydroxytryptamine (5,7-DHT) reduced the breakdown of the BBB and attenuated the brain 5-HT level without affecting the plasma 5-HT. Cyproheptadine, ketanserin (5-HT2 receptor antagonists) and vinblastine (a vesicular transport inhibitor) prevented the increased permeability of the BBB alone. The plasma and brain 5-HT continued to remain high. These observations suggest that (i) 5-HT plays an important role in the breakdown of BBB permeability in FS, (ii) this effect of 5-HT on BBB permeability is mediated by 5-HT2 receptors, and (iii) FS induced increase in BBB permeability is age dependent.

The 1992 study reported here assessed relationships between potable water supply and trash collection practices and the prevalence of dengue vector mosquito (Aedes aegypti) breeding sites in 30 towns located along the north coast of Venezuela. Within each study town, 100 homes were chosen. At each of these homes the number of water-bearing containers and containers harboring A. aegypti were determined and interviews were conducted to obtain information about the local water supply, trash collection services, and excreta disposal. In general, A. aegypti breeding indexes were high: 55% of the residences were found to harbor A. aegypti immature forms; there was an average of 118 breeding sites per 100 residences; and 24% of the water-bearing receptacles were observed to contain the mosquito. The statistical method of principal component analysis was employed to rank the 30 towns in terms of variables describing public service deficiencies, and correlations existing between the variables studied were determined. Direct correlations were found between two water supply variables (frequency and duration of water supply interruptions), between the excreta disposal and trash collection variables, between the duration of water supply interruptions and the Aedes breeding indexes, and between the duration of water supply interruptions and the mean number of A. aegypti breeding sites found in water storage containers. Overall, the towns with the poorest services were found to have the highest breeding indexes and the greatest numbers of water storage containers harboring the mosquito. It is concluded that public service (water supply and waste disposal) deficiencies were largely responsible for A. aegypti propagation in the study towns. Accordingly, it is recommended that local programs be implemented for recycling containers, constructing water storage tanks that cannot harbor Aedes larvae, and conducting health education and community participation campaigns directed against the mosquito.

In the retina of herring (Clupea harengus L.), rods are recruited from about 8 wk after hatching, and from this time there is a linear relationship between the number of rods in the photoreceptor cell population and the content of di22:6n-3 molecular species of phospholipids. Juvenile herring were reared from four weeks' post-hatching for 15 wk on either Artemia nauplii deficient in 22:6n-3 or on enriched Artemia nauplii containing 4.3% 22:6n-3. The visual performance of the fish was then determined at three light intensities (0.01, 0.1, and 1.0 lux) by observing their frequency of striking at live Artemia nauplii using infrared video recording. Herring reared on the diet containing no 22:6n-3 were less active predators, especially at the lowest light intensity where very few strikes were observed. The eyes of these fish contained greatly reduced levels of di22:6n-3 molecular species of total phospholipid, 2.1% vs. 12.0% in fish supplemented with 22:6n-3. The contribution of saturated and monounsaturated fatty acids in the molecular species of phosphatidylethanolamine (PE), phosphatidylserine (PS), and phosphatidylcholine (PC) was virtually unchanged, while 20:5n-3 and 22:5n-3 largely replaced 22:6n-3. There was an almost complete disappearance of di22:6n-3 PC, while the amounts of di22:6n-3 PE and PS fell by 18.1 and 20.6% to 2.7 and 7.6%, respectively. The dipolyunsaturated molecular species di20:5n-3, 20:5n-3/22:5n-3, and di22:5n-3 made up a substantial part of the deficit. We conclude that a dietary deficiency of 22:6n-3 during the period early in rod development impairs visual performance such that the fish can no longer feed at low light intensities.

To determine plasma catecholamine levels during the use of propofol for maintenance of anesthesia. We studied 20 randomly chosen patients undergoing elective orthopedic surgery under general intravenous anesthesia with propofol as the agent of maintenance along with pancuronium and fentanyl. Plasma concentrations of dopamine, noradrenalin and adrenalin were measured at the following times: baseline, after intubation and every 15 min during surgery. The levels obtained for the different samples were compared with baseline levels. At the doses used in our study, there were no significant differences in the way dopamine plasma levels varied. This was not the case with the other two hormones, however. Adrenalin levels were significantly higher at the first 3 extractions (post-intubation, 15 min and 30 min), and noradrenalin was significantly higher (p < 0.05) at all extraction times. Total intravenous anesthesia with propofol is associated with a significant increase in plasma concentrations of adrenalin and noradrenalin. Adrenalin returns to baseline levels 45 min after induction.

Boutonneuse fever (BF) is usually considered to be a benign rickettsiosis. However, severe presentations, resembling Rocky Mountain spotted fever, have been reported. There are few neurological complications, except in serious forms of the disease. We present a case of meningoencephalitis in a male adult suffering from BF, and we review the clinical, etiopathogenic and diagnostic aspects among the cases previously reported. We conclude that BF encephalitis would have the following characteristics: a) they mostly affect elderly males; b) they cause altered consciousness level, headaches and low rate of meningeal signs; c) absence of "tâche noir"; d) cerebrospinal fluid with slight pleocytosis and/or increased level of proteins, and normal glucose values; e) computerized tomography without significative changes, and f) high rate of morbidity and mortality. We emphasize that we should consider BF in the differential diagnosis of our environmental encephalitis.

A total of 16 new species records of Culicidae from Falcon State was collected at the "Juan Crisostomo Falcon National Park" (Sierra de San Luis), Natural Monument "Cerro Santa Ana", Coro, and La Vela. Species of Sabethini, Culicini and Toxorhynchitini Tribes were found in natural breeding sites (Phytotelmata), with special occurrence in plants belonging to Tillandsia, Vriesea, Guzmania, Aechmea (Bromelianceae), Heliconia (Heliconiaceae), Calathea (Marantaceae) and Colocasia (Araceae). Aedini and Mansonini were collected only as adults. A specie of Culex (Carrollia) was collected from an artificial container. The Culicidae species belong to 6 genera out of the 23 genera reported from Venezuela (Culex, Wyeomyia, Johnbelkinia, Aedes, Psorophora, Mansonia and Coquillettidia) and to 5 Tribes out of the 9 present in the country. The Aedini, Sabethini and Culicini Tribes were richer in species with 5, 4 and 4 species, respectively, than the Mansonini (2 species) and Toxorhynchitini (1 species) Tribes. We discuss some bioecological aspects regarding the 16 new-species records in Falcon State and give a checklist of the mosquito species previously reported in the literature.

Objective: To study the usefulness of several omeprazole regimens on gastric fluid volume and pH in patients undergoing elective surgery., Material and Methods: We analyzed 105 patients undergoing to elective surgery who received prophylactic treatment for the acid aspiration syndrome. They were randomly allocated into 7 homogeneous groups according to the therapeutic regime. Placebo (group 1), oral omeprazole (20 mg) the night prior to surgery (group 2), oral ranitidine (150 mg) the night before (group 6), two doses (the previous night and before surgery) in the remaining three groups: 20 mg of omeprazole (group 4), 40 mg of omeprazole (group 5) or 150 mg of ranitidine (group 7). In all patients we measured pH and volume of the gastric content after induction of anesthesia and one hour thereafter., Results: There were no statistically significant differences in mean pH values and gastric volume content among groups 1.2,3, and 6 (2.1 (pH) and 27 ml (gastric volume) in group 1, 2.1 and 23 ml in group 2, 2.6 and 19 ml in group 3, and 2.2 and 32 ml in group 6). With repeated doses of 40 mg omeprazol (group 5), mean pH values were comparable to those found with the two doses of ranitidine (4.1 +/- 1.8 vs 4.1 +/- 3.6) although gastric volumes were significantly less (12 +/- 2.6 ml vs 20 +/- 4.8 ml). These two groups showed significantly greater mean pH values and less gastric volumes than the remaining patients., Conclusions: The incidence of patients with gastric content deemed at risk for acid aspiration (pH less than 2.5 and gastric volume greater than 25 ml) was less after premedication with two oral doses of omeprazole (40 mg) than either two doses of ranitidine (150 mg) or smaller doses of both drugs.

An eight year old boy of first grade consanguinity parents, with important growth and mental retardation, generalized bone alterations and without corneal opacity nor mucopolysacchariduria is presented. All these features account on the syndrome of Dyggve-Melchior and Clausen.

In a group of 16 cases with adult-onset chronic proximal spinal muscular atrophy, we performed a morphometric analysis of 88 intramuscular nerves in biopsied biceps brachii muscles and correlated this analysis with clinical and histochemical parameters. The total number of large and small myelinated fibres in all fascicles was reduced to 71%, 23% and 83% of control values, respectively. The percentage reduction of large myelinated fibres for each case was significantly correlated with duration of illness, biceps muscle power and histochemical atrophy factors. Histograms of large intramuscular nerve fascicles showed unimodal distributions and shifts to the left with single peak increases at 1 micron. The densities of small myelinated fibres in large fascicles correlated with counts of 'enclosed' muscle fibres in ATPase preparations. Compared with amyotrophic lateral sclerosis, a lesser reduction of large myelinated fibres, but a greater increase in small myelinated fibres, was noted in adult-onset chronic proximal spinal muscular atrophy. These findings imply that there is a less marked loss of myelinated nerve fibres with more effective reinnervation in adult-onset chronic proximal spinal muscular atrophy than in amyotrophic lateral sclerosis.

Morphometric analysis of 90 intramuscular nerves from the biopsied biceps muscles of 16 cases of amyotrophic lateral sclerosis (ALS) were done and compared with controls. In all fascicles (large and small) of ALS, the total number and the numbers of large and small myelinated fibers were significantly reduced to 52, 16, and 64% of controls, respectively. A histogram of the largest fascicles revealed a unimodal distribution and a shift to the left, showing a single peak at 1 micron diameter. The results indicate proximal and distal involvement of the most remote portions of the nerves to the muscle in ALS and support previous suggestions of a neuronopathy.