Showing papers by "Oswaldo Cruz Foundation published in 1996"

A Schistosoma mansoni fatty acid-binding protein, Sm14, is the potential basis of a dual-purpose anti-helminth vaccine

Abstract: Molecular cloning of components of protective antigenic preparations has suggested that related parasite fatty acid-binding proteins could form the basis of the protective immune crossreactivity between the parasitic trematode worms Fasciola hepatica and Schistosoma mansoni. Molecular models of the two parasite proteins showed that both molecules adopt the same basic three-dimensional structure, consisting of a barrel-shaped molecule formed by 10 antiparallel beta-pleated strands joined by short loops, and revealed the likely presence of crossreactive, discontinuous epitopes principally derived from amino acids in the C-terminal portions of the molecules. A recombinant form of the S. mansoni antigen, rSm14, protected outbred Swiss mice by up to 67% against challenge with S. mansoni cercariae in the absence of adjuvant and without provoking any observable autoimmune response. The same antigen also provided complete protection against challenge with F. hepatica metacercariae in the same animal model. The results suggest that it may be possible to produce a single vaccine that would be effective against at least two parasites, F. hepatica and S. mansoni, of veterinary and human importance, respectively.

Ecological interactions of visceral leishmaniasis in the state of Bahia, Brazil

Abstract: The laboratory and field observations summarized in this paper on visceral leishmaniasis ecology in the State of Bahia, Brazil are based on the author's observations over the past 35 years in a number of state's foci, public health records and literature citations. The disease is endemic with epidemic outbreaks occurring every ten years and its geographical distribution is expanding rapidly in the last years. Leishmania chagasi is the main ethiologic agent of the visceral leishmaniasis but Le. amazonensis s. lato was the only leishmania isolated by other authors from some visceral leishmaniasis human cases in the state. Lutzomyia longipalpis (with one or two spots on tergites III and IV and two sized different populations) was epidemiologically incriminated as the main vector. It was found naturally infected with promastigotes, and it was infected with four species of leishmanias in the laboratory. Although the experimental transmission of Le. amazonensis by the bite of Lu. longipalpis to hamsters was performed, the author was not successful in transmitting Le. chagasi in the same way. The dog is the most important domestic source for infection of the vector, however it is not a primary reservoir. The opossum Didelphis albiventris was found naturally infected with Le. chagasi but its role as reservoir is unknown. Foxes and rodents were not found infected with leishmanias in Bahia.

VACTERL association, epidemiologic definition and delineation

Abstract: This study departed from a preconceived definition of VACTERL, including more than one of these six anomalies in the same infant:; V (vertebral anomalies), A (anal atresia), C (congenital heart disease), TE (tracheoesophageal fistula or esophageal atresia), R (reno-urinary anomalies), and L (radial limb defect). Under this definition, 524 infants were ascertained by ECLAMC from almost 3,000,000 births examined from 1967 through 1990. Observed association rates among VACTERL components as well as between VACTERL and other defects were compared against randomly expected values obtained from 10,084 multiply malformed infants (casuistic method) from the same birth sample. Conclusions were: 1) Cardiac defects are not a part of VACTERL. 2) Single umbilical artery, ambiguous genitalia, abdominal wall defects, diaphragmatic hernia, and anomalies that are secondary to VACTERL components (intestinal and respiratory anomalies, and oligohydramnios sequence defects) are frequent enough to be considered an “extension” of VACTERL, and cardiac defects should be included in this category. 3) Neural tube defects are negatively associated with VACTERL which could not be explained by selection bias or any other operational artifact. High embryonic lethality or mutually exclusive pathogenetic mechanisms could be suitable explanations. 4) Results were not clear enough to determine whether VACTERL should be defined by at least two or three component defects. © 1996 Wiley-Liss, Inc.

Patterns of detection of Strongyloides stercoralis in stool specimens: implications for diagnosis and clinical trials.

Abstract: Reported efficacies of drugs used to treat Strongyloides stercoralis infection vary widely. Because diagnostic methods are insensitive, therapeutic trials generally require multiple negative posttreatment stool specimens as evidence of drug efficacy. However, only a single positive stool specimen is usually required for study enrollment. To determine the reproducibility of detection of S. stercoralis larvae in the stool, 108 asymptomatic infected men submitted 25 g of fresh stool once a week for eight consecutive weeks for examination by the Baermann technique. During the 8-week study, 239 (27.7%) of 864 stool specimens were positive for S. stercoralis. Rates of detection of larvae in the stool specimens ranged from eight of eight specimens in 3 (2.8%) men to none of eight specimens in 36 (33.3%) men. Of 43 men for whom S. stercoralis was detected in at least two of the first four stool specimens, only 1 (2.3%) man tested negative on all of the next four specimens. In comparison, of 29 men who had detectable larvae in only one of the first four specimens, 22 (75.9%) tested negative on all of the next four samples. Thus, if these 29 men had been enrolled in a therapeutic trial between the first and second sets of four specimens, the efficacy of a drug with no activity against this parasite would have been estimated to be 76%. These data suggest that patterns of S. stercoralis detection vary widely among infected persons and that intermittent larval shedding can lead to inflated estimates of drug efficacy. Before a patient is entered in a clinical trial of drug efficacy, four consecutive stool specimens should be examined for S. stercoralis; only persons with two or more positive specimens should be enrolled.

Detection of extracellular proteases from microorganisms on agar plates

Abstract: We present herein an improved assay for detecting the presence of extracellular proteases from microorganisms on agar plates Using different substrates (gelatin, BSA, hemoglobin) incorporated into the agar and varying the culture medium composition, we were able to detect proteolytic activities from Pseudomonas aeruginosa, Micrococcus luteus and Serratia marcescens as well as the influence that these components displayed in the expression of these enzymes For all microorganisms tested we found that in agar-BHI or yeast extract medium containing gelatin the sensitivity of proteinase detection was considerably greater than in BSA-agar or hemoglobin-agar However, when BSA or hemoglobin were added to the culture medium, there was an increase in growth along with a marked reduction in the amount of proteinase production In the case of M luteus the incorporation of glycerol in BHI or yeast extract gelatin-agar induced protease liberation Our results indicate that the technique described here is of value for detecting extracellular proteases directly in the culture medium, by means of a qualitative assay, simple, inexpensive, straight forward method to assess the presence of the proteolytic activity of a given microorganism colony with great freedom in substrate selection

Are geoprocessing, environment, and health a possible combination?

Abstract: The use of geoprocessing techniques allows one to gather socioeconomic, health, and environmental data on a spatial basis. However, interpretation of associations between epidemiological and environmental variables requires the geoprocessing system design. The study scale and object choices precede conception of the system, conditioning the possible statistical and visual results. This scale must be compatible with the phenomenon on which one intends to focus, aiming at internal homogeneity and external heterogeneity of spatial units. The interdependency of spatial processes, reflected in the spatial configuration of social, environmental, and epidemiological data distribution, affects interpretation of causes for simultaneous processes. Geoprocessing allows for knowledge of the context or situational surroundings in which the damage to health takes place.

Comparison of the polymerase chain reaction with two classical parasitological methods for the diagnosis of Chagas disease in an endemic region of north-eastern Brazil.

Abstract: The sensitivities for Chagas disease diagnosis of haemoculture, xenodiagnosis, and polymerase chain reaction (PCR) amplification of Trypanosoma cruzi kinetoplast deoxyribonucleic acid (DNA) were compared for 101 patients living in an endemic region who were serologically positive for T. cruzi. PCR gave 60 positive results (59.4%), while a haemoculture was positive in 26 cases (25.7%) and xenodiagnosis in 36 (35.6%). Four xenodiagnosis-positive but PCR-negative patients were examined in detail. The discrepancies were not due to inhibition of the PCR reactions, as the samples were used successfully to amplify a human sequence. Nor were they due to a variation in kinetoplast DNA sequences, as the kinetoplast DNA of the parasite strains isolated from these patients after xenodiagnosis gave rise to the expected product when amplified by the PCR. We concluded that no parasite was present in the 5 mL of blood used for PCR, while probably a single T. cruzi cell was present in the blood volume ingested by the insects during xenodiagnosis (about 3 mL). This suggests that the total blood quantity collected for the PCR may be important with patients with low parasitaemia.

A cross-sectional serodiagnostic survey of canine leishmaniasis due to Leishmania chagasi.

Abstract: Jequie, a community of about 144,500 inhabitants located in the State of Bahia, Brazil, is endemic for both visceral and cutaneous leishmaniases. In the present epidemiologic study, the urban and inhabited periurban areas of the town were divided into 140 clusters of 0.25 km2 each. The seroprevalence of canine Leishmania antibodies was investigated using an enzyme-linked immunosorbent assay as a screening test since its sensitivity was significantly higher than that of an indirect immunofluorescence assay. A total of 1,681 dogs was surveyed in 34 randomly sampled clusters. The overall prevalence of Leishmania antibodies in the dog population was 23.5%, with intracluster prevalences ranging from 0% to 67%. There was no correlation of these seroprevalences with the intracluster densities of canine populations, or with the distances from individual clusters to the town center. Moreover, the Leishmania transmission did not seem to follow any clear-cut spatial pattern, since large disparities in the seroprevalences of contiguous clusters were found. Curiously, human cases of visceral leishmaniasis have never been observed in some clusters with a relatively high prevalence of canine seroprevalences. Eight parasite isolates from seropositive dogs were found to belong to the same serodeme and zymodeme as Leishmania (L.) chagasi. The implications of these findings with respect to the epidemiology and control of American visceral leishmaniasis are discussed.

Occurrence of living adult Wuchereria bancrofti in the scrotal area of men with microfilaraemia

Abstract: To determine the frequency with which living adult Wuchereria bancrofti can be detected by ultrasound in the scrotal area of men with filarial infection, we used a 7·5 MHz transducer to perform weekly ultrasound examinations on 100 microfilaraemic men (18–34 years old) from Greater Recife, Brazil. The peculiar pattern of movement that characterizes the adult worm image on ultrasound (the filaria dance sign) was detected in the lymphatic vessels of the spermatic cord in 80 men (bilaterally in 29 men). Among 20 men with no filaria dance sign, the geometric mean microfilarial density was 68/mL, compared with 238/mL and 775/mL among those with unilateral and bilateral filaria dance signs, respectively ( P = 0·0001). The lymphatic vessels of the spermatic cord appear to be a common, and perhaps the principal, site of adult W. bancrofti in men with asymptomatic microfilaraemia. Studies are needed to define the relationship between the presence of filarial worms in the scrotal area and the development of filaria-associated morbidity.

Identification of Single and Dual Infections with Distinct Subtypes of Human Immunodeficiency Virus Type 1 by Using Restriction Fragment Length Polymorphism Analysis

Abstract: The simultaneous presence of multiple HIV-1 subtypes has become common in communities with the growth of the pandemic. As a consequence, the potentiality for an increased frequency of HIV-1 mixed infections caused by viruses of distinct subtypes could be expected. Thus, there is a need to estimate the prevalence and geographic distribution of infections caused by viruses of a singular subtype as well as coinfections caused by two or more HIV-1 strains of distinct subtypes. To address this need, we have developed a genetic method based on restriction fragment length polymorphism (RFLP) to screen for these two types of infections within infected populations. In this assay, restriction enzymes may be used to predict the phylogroup of HIV-1 infected samples. A 297 bp pol fragment spanning the entire viral protease gene and a 311 bp fragment of the p24 gag region are used for this analysis. The viral regions are amplified by nested PCR using DNA templates from uncultured peripheral blood mononuclear cells (PBMC) or virus culture. Classification of HIV-1 strains to well defined subtypes B, D, F, and A/C is done by sequential endonuclease restriction analysis of a PCR amplified-protease gene followed by analysis of the p24 gag region. The electrophoretic migration patterns visualized by ethidium bromide staining or by radiolabeled probes are then determined on a 10% polyacrylamide gel. In infections caused by viruses of a singular subtype, a single restriction pattern is detected, whereas in multiple infections caused by two or more viral strains of different subtypes, the combination of different digestion patterns are observed in infected individuals. Using this methodology we have screened for genetic variations in HIV-1 proviral DNA from thirty-three Brazilian samples. Our RFLP procedure classified thirty-two samples as single infections caused by viruses of subtypes B (31) and F (1), and one sample as dual infection caused by distinct viral strains. Subsequent sequence and phylogenetic analysis of the viral protease gene in lymphocytes of all these patients confirmed our RFLP findings in single infections, and demonstrated the existence of two distinct HIV-1 strains of subtypes F and D in a patient which lymphocytes showed the simultaneous presence of two different digestion patterns. As up to now, single infections caused by subtype D variants were not identified in Brazil, our data provide the first evidence of subtype D HIV-1 in this country. Because sequencing of HIV proviral DNA is not particularly practical for large-scale molecular epidemiological studies, the protease/gag-based RFLP screening method will be useful to predict the phylogroup of HIV-1, and to identify multiple infections caused by HIV-1 strains of distinct subtypes. We believe that this information is crucial for both evaluation of the HIV-1/AIDS pandemic and intervention strategies.

Chagasic patients lack CD28 expression on many of their circulating T lymphocytes.

Abstract: A balanced host-parasite interaction during Trypanosoma cruzi infection allows for the establishment of a chronic infection that can last for many years. T cells are a major element responsible for parasite specific and non-specific immunity during the complex immune response of the host. However, the subpopulations of T cells involved in the response, as well as the exact mechanisms through which those cells are activated or rendered unresponsive, are not well defined. It is known that co-stimulatory signals, some of which are mediated via CD28, are of critical importance in the triggering of appropriate T cell responses. In this study the authors performed double-labelling studies to determine the frequency of expression of CD28 by CD4+ and CD8+ T lymphocytes in the peripheral blood of patients with Chagas' disease. The results show that chagasic patients throughout the spectrum of chronic clinical forms of the infection have significantly higher mean frequencies of CD4+CD28- and CD8+CD28-T cells, as compared with non-chagasic individuals. Considering the importance of CD28 for T-cell activation, the observed down-regulation or loss of CD28 during infection may indicate a possible basis for observed immunoregulatory events or distinct stages of T-cell activation in this infection. Recent evidence from patients with HIV/AIDS indicates that CD28- cell populations are more likely to undergo apoptosis, and increased apoptosis has been observed in experimental Chagas disease.

Thalidomide, a current teratogen in South America

Abstract: Thalidomide, mainly used for the treatment of leprosy, is a current teratogen in South America, and it is reasonable to assume that at present this situation is affecting many births in underdeveloped countries. Moreover, the potential re-marketing of thalidomide for the treatment of a large variety of diseases may extend the problem to the developed world. When the drug is available, the control of its intake during early pregnancy is very difficult since most pregnancies are unintended. The ongoing occurrence of thalidomide embryopathy cases went undetected by the ECLAMC, due to several factors: (1) low populational coverage through this monitoring system; (2) pre-existence of the teratogen with its effects present in both baseline (expected) and monitored (observed) materials; and (3) lack of a defined phenotype to be monitored. Thus, if thalidomide re-enters the market throughout the world, due to the wide range of new applications, occurrence of phocomelia alone might not be sufficient to detect its effects. By a case-reference approach, the ECLAMC registered 34 thalidomide embryopathy cases born in South America after 1965 whose birthplaces correspond to endemic areas for leprosy. Phocomelia was found in five of eleven fully described cases. Thus, phocomelia alone is neither specific nor sufficient to serve as a suitable phenotype to survey the teratogenic effects of thalidomide. Therefore, a thalidomide-like phenotype, defined as any bilateral upper and/or lower limb reduction defect of the preaxial and/or phocomelia types, should be included in the routine surveillance of birth defects in all programmes.

Gender differences in growth of school-aged children with schistosomiasis and geohelminth infection

Abstract: Light or moderate intensity infection with Schistosoma mansoni may contribute to growth deficits. We report on the effects of treatment for S. mansoni on growth and development in Brazilian schoolchildren. Anthropometric measurements were taken from 539 S. mansoni-infected children and their age- and sex-matched egg-negative controls between the ages of 7 and 15 years. The children as a whole exhibited chronic malnutrition, with growth retardation in height evident in 21% of the population. Infected children, however, were significantly smaller in height, weight, mid upper arm circumference (UAC), tricep skinfold (TSF), and subscapular skinfold (SSF) measurements than control children (P < 0.05). These differences were due primarily to a greater disparity between infected and egg-negative girls in height (P < 0.01), weight (P = 0.01), UAC (P = 0.O2), and TSF (P < 0.01). Nevertheless, girls demonstrated a better level of development and nutrition compared with boys. While infected boys were shorter and weighed less than controls, these differences were not significant. Growth and development in girls was negatively correlated with intensity of infection. Coinfection with S. mansoni and Trichuris appeared to act synergistically in the development of malnutrition.

Ultrasonographic assessment of the adulticidal efficacy of repeat high‐dose ivermectin in bancroftian filariasis

Abstract: Since diethylcarbamazine, the drug recommended for treatment of lymphatic filariasis, seems only partially effective against the adult worm, intense interest persists in identifying a macrofilaricidal drug for this infection. To evaluate directly in vivo the macrofilaricidal activity of repeat high-dose ivermectin, 15 men who had living adult Wuchereria bancrofti detected in the scrotal area by ultrasound were treated with 400 micrograms/kg of ivermectin at 2-week intervals for 6 months (total dose, 4.8 mg/kg). Serial ultrasound examinations were performed before, during, and for 6 months after treatment. Profound suppression of microfilaraemia followed the first dose of ivermectin, but movements characteristic of the adult worm on ultrasound remained unchanged both in location and pattern. Even when given in total doses of 4.8 mg/kg, ivermectin appears to have no observable activity against adult W. bancrofti, although its ability to suppress microfilaraemia makes it potentially useful for the control of lymphatic filariasis.

Tumor necrosis factor-α in human American tegumentary leishmaniasis

Abstract: Tumor necrosis factor-alpha (TNF-alpha) is a cytokine produced by activated macrophages and other cells. In order to verify whether the serum levels of TNF-alpha in American tegumentary leishmaniasis patients are associated with the process of cure or aggravation of the disease, 41 patients were studied: 26 cases of cutaneous leishmaniasis (CL) and 15 of mucocutaneous leishmaniasis (MCL). During active disease the serum levels of TNF-alpha of MCL patients were significantly higher than those of CL patients and control subjects (healthy individuals and cutaneous lesions from other etiologies). The MCL patients had serum titers of TNF-alpha significantly lower at the end of antimonial therapy than before therapy. After a six-month follow-up, the MCL patients had serum levels of TNF-alpha similar to those observed at the end of the therapy as well as to those of CL patients and control subjects. No significant variation in the serum levels of TNF-alpha was observed in CL patients throughout the study period (before, at the end of therapy and after a six-month follow-up). The possible relationship between the high TNF-alpha serum levels and severity of the disease is discussed.

The tegument of Schistosoma mansoni: functional and immunological features

Abstract: Some structural, biophysical and molecular features of the tegument are shown, highlighting their changes during maturation as important adaptative mechanisms for the survival of the parasite and resistance to immune attack. On the other hand, many antigens, targets of the immune response against the schistosome, are located on the tegument. Some of the features of these molecules are presented and the possibilities of using them in the immunological control of the disease are discussed.

Role of enterovirus 71 in acute flaccid paralysis after the eradication of poliovirus in Brazil.

Abstract: As a result of the successful initiative to eradicate poliomyelitis in the Americas, Brazil is now free of circulating wild poliovirus. The last cases of acute flaccid paralysis (AFP) with confirmed wild poliovi-rus isolation occurred in March 1989 (1). Since the elimination of wild poliovirus-associated poliomy-elitis in Brazil and the American region by intensive mass vaccination campaigns, acute paralytic illnesses from other causes constitute a greater proportion of suspected cases. Enterovirus 71 (EV71) can cause paralytic disease with persistent flaccid paralysis (which may be confused with paralysis caused by wild polioviruses); therefore, the presence of this enterovirus in a community may complicate the evaluation of poliomyelitis control progress. Se-rologic evidence of EV71 infections (demonstrated by neutralization assays) has been observed in AFP patients in Brazil (9). In addition, EV71 isolates have been obtained from patients with suspected cases of poliomyelitis and their healthy contacts from different areas in Brazil and from AFP patients in Peru and Bolivia (3, Ferreira et al., in preparation). EV71, the most recently recognized human en-terovirus in the family Picornaviridae, has been associated with outbreaks of hand-foot-and-mouth disease and central nervous system diseases (e.g., aseptic meningitis, encephalitis, and poliomyelitis-like paralytic diseases) with persistent or transient paralysis (2,4,5). Despite some understanding of the variability in clinical manifestations and epidemio-logic pattern, little is known about the contribution of EV71 to overall AFP in different countries. Immunoglobulin M (IgM) antibody to EV71 was measured by using an µ-capture enzyme immunoas-say (EIA) (6). The antigen was prepared from the BrCr strain of EV71, and biotinylated anti-EV71 monoclonal antibody as detector was also prepared (6). The results were expressed as the difference in mean optical density values measured in triplicate wells of positive antigen (P) and negative controls (N). A specimen was considered positive if the observed P-N was > 2 standard deviations (SD) above the mean of the optical densities for the negative control serum (P-N > 0.25). Serum samples were obtained from 92 infants from different regions of Brazil who had suspected poliomyelitis and symptoms of AFP during 1989 and 1990. The 138 samples included paired serum samples (S1 and S2) from 46 patients (92 sera) and 46 serum samples (S1 only) from 46 patients. Acute-phase serum samples (S1) were obtained 1 to 15 days after the onset of symptoms, and a second group of samples were obtained (S2) 15 to 45 days later; all were from children whose …

Structural variation in the glycoinositolphospholipids of different strains of Trypanosoma cruzi.

Abstract: The structures of the glycoinositolphospholipids (GIPLs) from five strains of the protozoan parasiteTrypanosoma cruzi have been determined. Two series of structures were identified, all but one containing the same Man4(AEP)GlcN-Ins-PO4 core. Series 1 oligosaccharides are substituted at the third mannose distal to inositol (Man 3) by ethanolamine-phosphate or 2-aminoethylphosphonic acid, as are some glycosyl-phosphatidylinositol-protein anchors ofT. cruzi. The core can be further substituted by terminal (1–3)-linked β-galactofuranose units. In contrast, Series 2 oligosaccharides do not have additional phosphorus-containing groups attached to Man 3, the latter being substituted instead by a single side chain unit of β-galactofuranose. Series 1 oligosaccharides are present in all strains (G, G-645, Tulahuen CL, and Y) whereas Series 2 structures are present mainly in CL and Y strains. The lipid moiety in the GIPLs from the G, G-645 and Tulahuen strains is predominantly ceramide, as reported for the Y strain, whilst that from the CL strain is a mixture of ceramide and alkylacylglycerol species. The lipid moiety of the GIPLs, and probably also the phosphoinositol-oligosaccharide structures may play an important immunomodulatory role in infection byT. cruzi.

Gram-chromotrope: a new technique that enhances detection of microsporidial spores in clinical samples.

Abstract: SUMMARY We have developed a new staining procedure that combines the traditional Gram staining for bacteria and the Weber's chromotrope staining method, the standard technique for the detection of microsporidia spores in clinical Specimens. This “Gram-chromotrope” staining technique enhances the staining characteristics of microsporidia spores and facilitates the easy detection and differentiation of spores from other microorganisms that are found in clinical specimens, especially stool samples. This new technique is fast, reliable, and simple to perform, and can be easily adapted for use in clinical laboratories.

Primary isolation of spotted fever group rickettsiae from Amblyomma cooperi collected from Hydrochaeris hydrochaeris in Brazil

Abstract: This paper reports the first isolation of a spotted fever group rickettsia from an Amblyomma cooperi ixodid collected from a capybara (Hydrochaeris hydrochaeris) in an endemic area of spotted fever in the County of Pedreira, State of Sao Paulo, Brazil. Isolation was performed in Vero cell culture and submitted to immunofluorescence, using antibody from Rickettsia rickettsii-positive human serum.