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L V Kirchhoff

Publications and source records attributed to L V Kirchhoff.

At least 55 records · Page 3Linked to original sources
PubMed1989

Trypanosoma cruzi expresses diverse repetitive protein antigens.

We screened a Trypanosoma cruzi cDNA expression library with human and rabbit anti-T. cruzi sera and identified cDNA clones that encode polypeptides containing tandemly arranged repeats which are 6 to 34 amino acids in length. The peptide repeats encoded by these cDNAs varied markedly in sequence, copy number, and location relative to the polyadenylation site of the mRNAs from which they were derived. The repeats were specific for T. cruzi, but in each case the sizes of the corresponding mRNAs and the total number of repeat copies encoded varied considerably among different isolates of the parasite. Expression of the peptide repeats was not stage specific. One of the peptide repeats occurred in a protein with an Mr of greater than 200,000 and one was in a protein of Mr 75,000 to 105,000. The frequent occurrence and diversity of these peptide repeats suggested that they may play a role in the ability of the parasite to evade immune destruction in its invertebrate and mammalian hosts, but the primary roles of these macromolecules may be unrelated to the host-parasite relationship.

Amino Acid Sequence↗
PubMed1989

Detection of Trypanosoma cruzi by DNA amplification using the polymerase chain reaction.

The polymerase chain reaction was used to amplify a 188-base pair (bp) segment of the repetitive 195-bp nuclear DNA sequence of Trypanosoma cruzi that is the most abundant sequence in this organism. The reaction amplified this repetitive element in four T. cruzi isolates from widely separated geographic regions. No amplification of the 188-bp fragment occurred when DNAs extracted from Leishmania spp., African trypanosomes, or blood samples from mice and humans were used. Amplification of one-half of the DNA from a single T. cruzi parasite produced an amount of the 188-bp element that was readily visible in a gel stained with ethidium bromide. Hybridization of a radiolabeled probe to membrane-bound amplification products increased the sensitivity to a level at which 1/200 of the DNA in a single parasite could be detected. T. cruzi DNA was readily detected in DNA extracted from the abdominal contents of infected insect vectors reared in the laboratory. No parasite DNA was detected in the blood samples of two individuals known to be infected with T. cruzi, possibly because in such patients the number of circulating parasites are extremely low or because parasitemias are intermittent. These results represent a considerable increase in sensitivity over previously reported methods for the detection of T. cruzi infections. Polymerase chain reaction amplification can be used to evaluate large numbers of samples in a single day and thus should be useful in large-scale studies of the prevalence of T. cruzi in both insect vectors and mammalian hosts.

Animals↗
PubMed1989

Molecular cloning of mtp70, a mitochondrial member of the hsp70 family.

We have isolated a gene from the protozoan parasite Trypanosoma cruzi that encodes a previously unidentified member of the 70-kilodalton heat shock protein (hsp70) family. Among all the eucaryotic hsp70 proteins described to date, this trypanosome protein, mtp70, is uniquely related in sequence and structure to the hsp70 of Escherichia coli, DnaK, which functions in the initiation of DNA replication. This relationship to DnaK is especially relevant in view of the intracellular location of the protein. Within the trypanosome, mtp70 is located in the mitochondrion, where it associates with kinetoplast DNA (kDNA), the unusual mitochondrial DNA that distinguishes this order of protozoa. Moreover, mtp70 is located in the specific region of the kinetoplast in which kDNA replication occurs. In view of the known functions of DnaK, the localization of mtp70 to the site of kDNA replication suggests that mtp70 may participate in eucaryotic mitochondrial DNA replication in a manner analogous to that of DnaK in E. coli.

Amino Acid Sequence↗
PubMed1988

Ubiquitin genes in trypanosomatidae.

A ubiquitin encoding cDNA from Trypanosoma cruzi, the protozoan cause of Chagas' disease, was isolated by immunoscreening a lambda gt11 expression library with serum from a mouse chronically infected with this parasite. The cDNA encodes a precursor protein consisting of four tandem repeats of ubiquitin differing from that of Saccharomyces cerevisiae in four positions, followed by an unrelated 52-amino acid tail containing a putative metal and nucleic acid binding domain. Southern and Northern blots of DNA and RNA from various strains of T. cruzi and from several African trypanosome and Leishmania isolates revealed dramatic differences in the numbers and sizes of ubiquitin genes and transcripts. One of the T. cruzi isolates examined has as many as 10 ubiquitin genes, while a strain of Leishmania donovani appears to have only 1. Forty or more tandemly arranged ubiquitin coding repeats are present in some genes, while others have only two or three. Evidence for stage-specific expression of a ubiquitin transcript was found in one strain, but no stress-induced changes in the pattern of transcripts were detected in the one isolate examined. Thus the cellular requirements for ubiquitin in trypanosomatids can be supplied by diversely organized genes containing highly variable numbers of ubiquitin coding repeats.

Amino Acid Sequence↗
PubMed1987

Trypanosoma cruzi exhibits inter- and intra-strain heterogeneity in molecular karyotype and chromosomal gene location.

Molecular karyotypes of 6 strains and 6 clones of Trypanosoma cruzi were determined using orthogonal-field-alternation gel electrophoresis. At least 15 different chromosome-sized DNA molecules, ranging in size from less than 200 kilobase pairs to greater than 2000 kilobase pairs, were resolved for each of the isolates examined. Many of the bands were present in different relative intensities suggesting that the number of individual chromosomes per organism may be considerably higher. Significant inter- and intra-strain differences in molecular karyotype and in the chromosomal locations of the genes for the spliced leader, tubulins, 5S ribosomal RNA and a heat shock protein were found. These marked chromosomal differences among T. cruzi strains and clones may be related to the high degree of phenotypic heterogeneity previously found in this parasite.

Animals↗
PubMed1987

American trypanosomiasis (Chagas' disease) in Central American immigrants.

A survey was conducted to determine the prevalence of infection with Trypanosoma cruzi, the protozoan etiologic agent of American trypanosomiasis (Chagas' disease), among Nicaraguan and Salvadoran immigrants living in the Washington, D.C., area. The serum samples of study subjects were tested for reactivity with T. cruzi antigens in an enzyme-linked immunosorbent assay, and also tested for antibody specific for the 72 and 90 kilodalton (kDa) surface glycoproteins of the parasite in an immunoprecipitation and electrophoresis procedure. Xenodiagnosis using reduviid bugs to detect parasites, and clinical evaluations for cardiac and gastrointestinal disease were performed in patients in whom results of both serologic tests were positive. Of 205 subjects studied, 4.9 percent were infected with T. cruzi, and parasites were isolated from 50 percent of those in whom xenodiagnosis was attempted. No significant cardiac or gastrointestinal abnormalities were detected in the six infected patients who were evaluated clinically. These findings suggest that a sizable proportion of persons in this immigrant group are infected with this organism. Thus, routine serologic testing for antibody to T. cruzi may be warranted in immigrants from these countries, especially in view of the potentially serious consequences of infection with this parasite, and also because of the risk of transmission of T. cruzi by blood transfusion.

Chagas Disease↗
PubMed1986

Treatment of Hodgkin's disease stages I and II with chemotherapy alone.

Ten previously untreated patients with stages I and II Hodgkin's disease were treated with MOPP chemotherapy alone. Eight of nine evaluable patients went into complete remission (CR). Six remained in CR on the average 82.7 months after induction. Two patients relapsed early and died more than 5 years after entering the study. The patient in whom CR was not achieved died as well. The estimated 10-year survival rate is 51%. This experience adds to the very small number of reports that have appeared describing the results of treatment of patients with early stage Hodgkin's disease with MOPP alone.

Adolescent↗
PubMed1985

Chagas' disease in Latin American immigrants.

Chagas' disease is caused by Trypanosoma cruzi, a protozoan parasite that infects an estimated 10 to 12 million Latin Americans. Because infection is often lifelong, individuals from endemic areas may develop symptomatic chronic Chagas' disease years after emigrating to the United States. We report three immigrant patients from Latin America who illustrate the indeterminate phase and the symptomatic cardiac and gastrointestinal syndromes of this illness.

Adult↗
PubMed1985

gp72, the 72 kilodalton glycoprotein, is the membrane acceptor site for C3 on Trypanosoma cruzi epimastigotes.

We examined the interaction of complement component C3 with surface molecules on Trypanosoma cruzi. Five- to six-fold more C3 was bound to epimastigotes (Epi) than to metacyclic trypomastigotes (CMT) of strain M88. Epi and CMT were surface iodinated, then incubated in C8-deficient serum, and detergent lysates were applied to anti-C3 antibody that had been coupled to Sepharose. We found that 9.20-10.24% of applied 125I-Epi protein bound to anti-C3-sepharose, compared to 2.64% binding of 125I-CMT protein. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis showed that C3 was attached to 125I-Epi protein by a covalent bond. Samples eluted from anti-C3-sepharose with hydroxylamine revealed a single, major, 72 kD band, suggesting that C3b attaches almost exclusively to the 72 kD glycoprotein of Epi by a hydroxylamine-susceptible ester bond. An antiserum was prepared from lysates of serum-treated Epi that had been affinity-purified on anti-C3-sepharose. This antiserum immunoprecipitated a single 72 kD component (gp72) from surface-iodinated Epi, and specifically recognized only gp72 from Epi in immunoblots. In contrast to the results with Epi, gp72 on CMT was not found to be an efficient acceptor molecule for C3 deposition. The results are the first to evaluate the acceptor site for C3 deposition on a parasite, and they show that gp72 on Epi, but not gp72 on CMT, serves as the preferential acceptor for C3 during antibody-independent alternative complement pathway activation.

Adult↗
PubMed1985

Feasibility and efficacy of in-home water chlorination in rural North-eastern Brazil.

The purpose of this study was to assess the feasibility, acceptability and effect of an in-home water chlorination programme in a rural village. Previous studies at this site showed high levels of faecal coliforms in household water, high diarrhoea rates in children, and enterotoxigenic Escherichia coli and rotaviruses were the most common pathogens isolated from patients. Household water came from a pond and was stored in clay pots. No homes had sanitary facilities. A blind, cross-over trial of treatment of household water with inexpensive hypochlorite by a community health worker was carried out over 18 weeks among 20 families. Water in the clay pots was sampled serially, and symptom surveillance was done by medical students. The programme was generally acceptable to the villagers and no change in water use patterns were apparent. The mean faecal coliform level in the chlorinated water was significantly less than in the placebo treated samples (70 vs 16000 organisms/dl, P less than 0.001). People living in houses receiving placebo treatment had a mean of 11.2 days of diarrhoea per year, and the highest rate of 36.7 was among children less than 2 years old. Diarrhoea rates were not significantly different among the participants while exposed to water treated with hypochlorite. We conclude that a low-cost programme of this type, which utilizes community resources, is logistically feasible, appears to be culturally acceptable in this setting, and can result in a marked reduction in water contamination. The lack of effect on diarrhoea rates suggests that improvement in water quality may affect morbidity only when other variables relating to faecal-oral agent transmission are ameliorated at the same time.

Adult↗
PubMed1985

Epidemiology and clinical significance of blood cultures positive for coagulase-negative staphylococcus.

Coagulase-negative staphylococci are frequently isolated from blood cultures. As these organisms may occasionally cause serious disease, differentiating bacteremia from contamination is very important but often difficult. Over a 26-month period, of 29,542 blood cultures processed at the University of Michigan Medical Center, 2,875 (9.7%) were positive, and of those, 694 (from 527 patients) grew coagulase-negative staphylococci. Isolates from the 439 patients with only one blood culture positive for coagulase-negative staphylococci and those from the 18 patients with two positive cultures 10 days or more apart were deemed contaminants. Review of the records of the remaining 70 patients with multiple isolates indicated that 33 had had an episode of true bacteremia, 29 (87.9%) of which were associated with intravascular catheters or prosthetic valves. Overall, 85% of all coagulase-negative staphylococci isolated during the study period were judged to be contaminants. Seventy-one percent of the blood cultures drawn during the episodes of bacteremia were positive for coagulase-negative staphylococci as opposed to only 34% in the patients with contaminated cultures (p less than 0.01). Moreover, coagulase-negative staphylococci grew in both aerobic and anaerobic bottles in 85% of blood culture sets drawn during episodes of bacteremia, but in only 30% of the cultures thought to be contaminated (p less than 0.001). Growth of coagulase-negative staphylococci in less than 48 hours was also significantly associated with bacteremia (p less than 0.01). Antibiotic sensitivity patterns were not useful in differentiating bacteremia from contamination.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1985

Molecular mimicry of a carbohydrate epitope on a major surface glycoprotein of Trypanosoma cruzi by using anti-idiotypic antibodies.

The use of anti-idiotypic antibodies (Ab2) to induce anti-microbial immunity might be particularly advantageous with respect to responses directed against carbohydrate determinants, because it may not be feasible to reproduce these epitopes by recombinant DNA technology. In the present studies, rabbit Ab2 were produced against a recurrent BALB/c idiotype defined by a monoclonal antibody (WIC 29.26) with specificity for a carbohydrate epitope of a major surface glycoprotein of Trypanosoma cruzi. The Ab2 induced specific antibodies in mice, rabbits, and guinea pigs, and reacted with parasite-induced anti-T. cruzi antibodies from mice and rabbits as well as humans. The behavior of this Ab2 is therefore consistent with that of the antigen itself, and suggests that molecular mimicry of carbohydrate epitopes can be easily achieved.

Animals↗
PubMed1984

Strains and clones of Trypanosoma cruzi differ in their expression of a surface antigen identified by a monoclonal antibody.

Four Trypanosoma cruzi strains and 50 clones derived from a total of 10 strains were assayed with a monoclonal antibody, WIC 29.26 Ab, for expression of an epitope previously demonstrated to be on the carbohydrate portion of a 72 000 Da surface glycoprotein of Y strain epimastigotes. WIC 29.26 Ab bound to only 2 of the 4 strains and 23 of the 50 clones tested. A group of 10 cloned isolates from one strain contained both reactive and non-reactive clones. Competitive inhibition studies with soluble extracts of the reactive and non-reactive isolates suggested that in addition to being absent from the surface membrane, the antigenic determinant is not synthesized by the non-reactive parasites. These data indicate that the epitope recognized by WIC 29.26 Ab is not present on all T. cruzi epimastigotes, and provide the first demonstration of clone-specific differences in a parasite antigen detected by a monoclonal antibody. No correlation was found between the reactivity of isolates with WIC 29.26 Ab and the previously investigated parameters of growth rate and modal volume.

Animals↗
PubMed1984

A case of schistosomiasis japonica: resolution of CAT-scan detected cerebral abnormalities without specific therapy.

A 28-year-old Marine Corps officer developed Katayama fever with central nervous system (CNS) manifestations 6 weeks after swimming on Leyte Island in the Philippines. Symptoms consisted of fever, nausea and vomiting, focal visual field deficits and mild confusion. CAT-scan of the patient's head initially revealed multiple lucencies and severe edema in the left frontal, parietal and occipital lobes. No schistosome eggs were found in the patient's stool, and therefore he was treated with a 10-week course of dexamethasone with resolution of all symptoms over 3 months. Repeat CAT-scan after symptoms cleared showed complete resolution of the focal abnormalities seen earlier. The diagnosis was subsequently established by positive serology and by finding eggs in the patient's stool and in tissue obtained by liver biopsy. This is the first report of CAT-scan-detected focal CNS lesions in a patient with acute schistosomiasis japonica, and resolution of the CNS abnormalities, temporally related to non-specific steroid treatment, is documented as well.

Adult↗
PubMed1984

Cryptic epitope explains the failure of a monoclonal antibody to bind to certain isolates of Trypanosoma cruzi.

A mouse monoclonal antibody, WIC 29.26 Ab, has previously been characterized as recognizing a carbohydrate epitope on a 72,000 m.w. glycoprotein (GP72) expressed on the surface of Trypanosoma cruzi epimastigotes and metacyclic trypomastigotes. This molecule has been implicated as a receptor in the control of parasite transformation, and when used as an immunogen in mice, partially protects against T. cruzi infection. In previous experiments in which a radioimmunoassay was used, WIC 29.26 Ab was found to react with approximately 50% of T. cruzi strains and clones derived from a variety of sources. In this study, we attempted to determine whether the WIC 29.26 Ab-nonreactive isolates lack the entire GP72 or merely lack the epitope recognized by this monoclonal antibody. WIC 226.4 Ab, a monoclonal antibody raised against periodate-treated GP72, reacted in an immunofluorescence assay with all strains and clones studied, including those which had not reacted with WIC 29.26 Ab. Likewise, two polyvalent rabbit sera, directed specifically against GP72, bound to all T. cruzi isolates tested. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis of detergent lysates of surface-labeled epimastigotes immunoprecipitated with WIC 29.26 Ab showed that the epitope bound by this antibody was present in all but one of the parasites that were surface-nonreactive, as well as in all those that were surface-reactive. WIC 29.26 Ab precipitated a single 72K Mr band from most strains and clones, but in several cases 79K Mr and 66K Mr bands were seen. Isolates from both the surface-reactive and the surface-nonreactive groups showed the latter pattern. These results demonstrate that GP72, or similar electrophoretic variants--and with one exception, the carbohydrate epitope bound by WIC 29.26 Ab--are present in the surface membrane of all strains and clones tested. This observation suggests that in intact epimastigotes of the surface-nonreactive isolates, the epitope is not accessible because of structural changes in the molecule itself or because of differences in the membrane environment of GP72.

Animals↗
PubMed1983

Prospective study of diarrheal illnesses in northeastern Brazil: patterns of disease, nutritional impact, etiologies, and risk factors.

Diarrhea is a leading cause of death in tropical countries. One of the highest childhood mortalities is in northeastern Brazil, where little is known about the morbidity, etiology, and risk factors of diarrhea. Prospective village surveillance over 30 months revealed diarrhea attack rates of more than seven episodes per child-year at six to 11 months of age among the children of the poorest families. Other risk factors included early weaning and the lack of toilets. Diarrhea led to weight loss and stunted growth. Enterotoxigenic Escherichia coli and rotaviruses were the most common pathogens, accounting for 21% and 19% of cases, respectively, followed by Shigella species (8.0%), Campylobacter jejuni (7.5%), Giardia species (6.7%), Strongyloides species (5.3%), and enteropathogenic E coli serotypes (4.6%). Most (84%) enterotoxigenic E coli were isolated during the rainy season of October to March (P less than 0.03), whereas 71% of rotaviral illnesses occurred during the drier months of June to October (P less than 0.03). In the present study, the early occurrence and nutritional impact of diarrhea and weaning, as well as the major etiologic agents of diarrhea and their different seasonal patterns have been defined for this region in which life-threatening diarrhea is endemic.

Adolescent↗
PubMed1983

Incubation in mice provides a signal for the differentiation of Trypanosoma cruzi epimastigotes to trypomastigotes.

The differentiation of Trypanosoma cruzi epimastigotes into trypomastigotes was studied in diffusion chambers subcutaneously implanted in mice. Using epimastigotes of the Tulahuén strain, transformation was first evident at 16 h after implantation and reached its maximum (92% trypomastigotes) by 24 h. Shortly before their differentiation into trypomastigotes, epimastigotes were found to develop resistance to lysis by the alternative pathway of complement. Furthermore, implantation of stationary-phase (as opposed to log-phase) parasites resulted in the accumulation of large numbers of complement-resistant epimastigotes in the chambers. These observations suggest that epimastigotes pass through a complement-resistant transitional stage before differentiating into trypomastigotes and that transformation may require cell division. In a further series of experiments, epimastigotes recovered 7 h after implantation in mice were found to differentiate into trypomastigotes when cultured in vitro for an additional 17 h at 37 degrees C. This observation indicates that the events which trigger the morphologic transformation of epimastigotes into trypomastigotes can be dissociated operationally from the differentiation process itself.

Animals↗