PubMed Health⌕ Search

Biomedical subjects

M Lorca

Publications and source records attributed to M Lorca.

At least 19 recordsLinked to original sources
PubMed2001

Highly diastereofacial anti-aldol reaction: practical synthesis of optically active anti-2-alkyl-3-hydroxycarboxylic acid ester units.

A variety of esters derived from commercially available norephedrine were used in diastereoselective anti-aldol reactions. The aldol reaction of designed 2-(N-2-methylbenzyl-N-2,4,6-trimethylbenzyl)amino-1-phenylpropanol esters 4a-d with aldehydes furnished anti-2-alkyl-3-hydroxycarboxylic acid esters in excellent diastereomeric ratios (>98:2) when LDA-Cp2ZrCl2 (0.3 equiv) was used for enolization, followed by transmetalation into the zirconium enolate for aldolization. The novel auxiliary 3 for the anti-aldol reaction does not exhibit the ordinary basicity of tertiary amines; 3 can be extracted from acidic media with organic solvents. Its use is, therefore, very advantageous not only for extraction of the aldol products from the acidic water solutions, but also for recovering the chiral auxiliary 3 after the reductive cleavage. Treatment of aldol or 3-protected aldol products with DIBAL-H or LiAlH4 affords the versatile synthons, 2-alkyl-propane-1,3-diols or those 3-protected diols in >98% ee's together with 3 in nearly quantitative recovery.

Journal Article↗
PubMed2001

Evaluation of a Triatoma infestans elimination program by the decrease of Trypanosoma cruzi infection frequency in children younger than 10 years, Chile, 1991-1998.

Chagas disease is widespread in Chile, distributed in rural and periurban areas in the 7 most northern regions of the country. The principal vector of Trypanosoma cruzi is Triatoma infestans. The interruption of the domestic cycle of transmission of T. cruzi has been attempted by health education, human housing improving, and elimination of the vector by means of systematic insecticide spraying of human dwellings. Spraying with insecticides has been supported by Chile's health authorities and has been carried out for the last 12 years. A total of 13,280 children (aged up to 10 years) were randomly selected from 47 counties in the area of Chile endemic for Chagas disease, and blood samples were collected to determine the levels of antibodies to T. cruzi by indirect hemagglutination and enzyme-linked immunosorbent assay tests. The results of the tests were analyzed to determine the changes that occurred after 12 years of insecticide spraying of dwellings to eliminate T. infestans infestation. A total of 142 (1.1%) samples of children showed antibodies to T. cruzi. This rate is significantly lower than the data generated in similar studies conducted in 1982-1985. The following reduction in prevalence rates were observed in each of the 7 endemic regions of the country: region I, 5.5-0.3%; region II, 6.6-0.3%; region III, 9.8-1.0%; region IV, 7.2-2.0%; region V, 5.2-1.9%, Metropolitan region, 1.4-0.6%; and region VI, 1.4-0.4%. Serovigilance of T. cruzi antibodies level represents a novel approach that may allow the evaluation of the impact of the vector elimination program. The results identify regions that need to strengthen the efforts to reduce the insect infestation of dwellings.

Animals↗
PubMed2001

[Serological certification of the interruption of the vectorial transmission of Chagas disease in Chile].

BACKGROUND: In 1999 an International Commission of Experts evaluated the sanitary interventions that started in 1980 to eliminate Triatoma infestans, the biological vector of Chagas disease, to certify if the conditions needed to interrupt the transmission, were achieved. AIM: To report the data used by the International certification commission to certify the interruption of vectorial transmission of Chagas disease in Chile. MATERIAL AND METHODS: A comparative evaluation of the prevalence of Trypanosoma cruzi antibodies in children surveyed between 1994 and 1995 and between 1995 and 1999. RESULTS: In the first period, 5,948 children were surveyed and in the second, 5,069 children were studied. Twenty children (0.4%) were positive for Trypanosoma cruzi antibodies in the second survey. These figures were significantly lower than the 1.1% prevalence detected between 1994 and 1995. In only three of the 20 cases, the transmission through vectors was confirmed, which represents a 99.4% reduction of this way of transmission. CONCLUSIONS: Based on these findings, the Commission certified that Chile is the second country in Latin America to interrupt the vector transmission of Chagas disease. The successful public health program for Chagas disease will have a positive impact improving the quality of life of rural populations, that are most exposed to the disease.

Animals↗
PubMed2000

Development of an immunoenzymatic assay for the detection of human antibodies against Trypanosoma cruzi calreticulin, an immunodominant antigen.

We have developed an indirect immunoenzymatic assay (ELISA) for the detection of human antibodies against calreticulin (formerly known as Tc45), a dimorphic Trypanosoma cruzi antigen, described in our laboratory. PVC microtitration plates were sensitized with the monoclonal anti-calreticulin antibody (MoAb) and reacted with calreticulin present in a partially purified preparation. The presence of anti-T. cruzi calreticulin IgG in sera from infected individuals was tested. The data generated with this assay were validated by correlation, in a regression analysis, with those obtained by an indirect immunoradiometric assay (IRMA). From the 12 seropositive sera (as defined by a commercial test), eight came out positive and four negative in both assays. The 12 human sera were also analyzed in direct immunometric assays (ELISA and IRMA), where the solid phase was sensitized with a whole parasite extract. The direct ELISA and IRMA correlated positively (P<0.01). Further validation of this ELISA was achieved with an indirect immunofluorescense assay. The high degree of significance obtained when the indirect IRMA and ELISA systems were compared, indicated that the relatively small sample number used (12) was statistically satisfactory for the purposes of this investigation. Thus, the IRMA can be replaced by the ELISA, with advantages mainly derived from the cumbersome manipulation of radioactive wastes. The MoAb used as an antigen capture agent in the ELISA proposed here, recognizes a homologous protein in Trypanosoma rangeli, suggesting that individuals infected with this parasite might have crossreactive antibodies. However, the system retains its diagnostic interest, given the facts that the MoAb does not recognize a homologous protein in Leishmania mexicana, Leishmania donovani, or Crithidia fasciculata.

Animals↗
PubMed1999

Seroprevalence of Toxoplasma gondii infection in sheep and alpacas (Llama pacos) in Chile.

Serum samples from 408 sheep from different regions of Chile and 447 alpacas (Llama pacos) from the north of the country were tested for Toxoplasma gondii antibodies. The indirect haemagglutination test (IHAT) was used in both species and the indirect immunofluorescence test (IIFT) was also used on the sheep samples in order to compare the performance of the tests in that species. In both tests, titers > or = 1:16 were considered diagnostically significant. Sera from 49 sheep (12%) were positive to T. gondii antibodies by the IHAT. When using the IIFT, 114 sheep sera (28%) were positive. The different results obtained in sheep sera between the tests were significant (p < 0.0001). No differences were observed between geographical locations or sex of the sampled sheep regarding serological detection of T. gondii antibodies in sheep. As expected, adult sheep showed higher T. gondii reactivity than young sheep (p = 0.0008). The corrected prevalence of toxoplasmosis in alpaca was 16.3% (32 positive out of 447). The rather low prevalence in alpacas may be associated with their extensive management as well as the extreme climatic conditions of The Andes which apparently would not be favorable for the transmission of the parasite.

Animals↗
PubMed1997

[Clinical and epidemiological study of blood donors seropositive for Trypanosoma cruzi].

BACKGROUND: The prevalence of positive antibodies for Chagas disease in a blood bank of a public hospital in Santiago is 1.2%. These positive individuals should be informed about their condition and further studied. AIM: To perform a serological confirmation, an epidemiological and clinical assessment of blood donors seropositive for Chagas disease in a blood bank of Santiago. SUBJECTS AND METHODS: One hundred eleven seropositive donors, detected between 1994 and 1996, were studied. Serological reactions were confirmed with ELISA and indirect immunofluorescence reactions. Those confirmed as positive were subjected to a clinical-epidemiological questionnaire, physical examination, EKG and barium enema or swallow according to symptoms. RESULTS: Seventy individuals were confirmed as positive for Chagas disease. Most of them lived at least once in an endemic zone for Chagas disease or in a sun-dried clay brick house. Forty-percent of individuals knew reduviid bugs and 14% had been bitten by these insects. Twenty six percent of these subjects had an abnormal EKG (61% with bradycardia). No digestive diseases attributable to Chagas disease were detected. CONCLUSIONS: The high prevalence of problems attributable to Chagas disease among seropositive blood donors underscores the need of an epidemiological follow up of these individuals.

Adult↗
PubMed1996

A putative pyruvate dehydrogenase alpha subunit gene from Trypanosoma cruzi.

A full-length DNA clone encoding a putative pyruvate dehydrogenase alpha subunit (E1 alpha) gene was isolated from a Trypanosoma cruzi (RA strain) DNA library. Sequencing of this clone revealed it to encode a 378 amino acid protein (M(r) 42774) with high sequence similarity to E1 alpha obtained from different sources. The highest score is obtained with human E1 alpha: 43,3% similarity. Southern blot analysis is consistent with the existence of a single copy of this putative T. cruzi E1 alpha gene per haploid genome in different parasite strains. Expression of this gene was demonstrated by Northern blot analysis and its trans-splicing acceptor site was identified by Polymerase Chain Reaction-mediated amplification of its cDNA.

Amino Acid Sequence↗
PubMed1995

Diagnostic value of detecting specific IgA and IgM with recombinant Trypanosoma cruzi antigens in congenital Chagas' disease.

The present study compares the early diagnosis of congenital Chagas' disease with a DOT assay using recombinant antigens with immunofluorescence antibody testing (IFAT) and an enzyme-linked immunosorbent assay (ELISA). The studies were performed using cord blood and sera of 12 infected newborns (group I) and 12 uninfected ones born to Trypanosoma cruzi-infected mothers (group II). Conventional IFAT and ELISA showed positive results for IgG at high titers, in infants and mothers of both groups; IgA antibodies were detected by ELISA in four of the infected infants and IgM was detected in two of them. All sera of the uninfected infants were negative for IgA and IgM in the ELISA. Application of a DOT assay using eight recombinant T. cruzi antigens allowed detection of specific IgA in the cord blood of six of the infected cases and IgM in eight of them. Repetition of these serologic tests in samples obtained during a monthly follow-up gave positive results for IgA in two of the initially negative infants of group I and for IgM in four of them. This means that diagnosis of congenital T. cruzi infection was confirmed, through demonstration of specific IgM, in all infected infants, and of IgA in eight of them. The importance of late detection of IgM in siblings born of infected mothers is discussed. The detection of IgM and IgA in sera obtained after birth is believed to be due to a congenital transmission of the parasite that occurred late in pregnancy. No IgA or IgM antibodies could be detected by the DOT assay in the sera of the negative controls.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1994

[Highly homogeneous Trypanosoma cruzi populations in a low endemic region for Chagas disease: importance in the pathogenesis of Chagas disease in Chile].

Trypanosoma cruzi from 23 chronic and acute cases of Chagas disease were isolated and further characterized. They displayed a remarkable similarity, after typification by schizodeme analysis using restriction endonuclease Eco Ri. All T cruzi isolates belonged to the isoenzymatic Zymodeme 2 Bol. These findings were corroborated analyzing cases from families with demonstrated congenital transmission. The acute cases were newborns or offspring of infected mothers or patients with AIDS. All T cruzi strains isolated in these cases were very similar with almost identical schizodemes. Patients with AIDS had severe clinical manifestations whereas children with congenital infections had a great variety of clinical symptoms. In chronic cases, little clinical differences were observed for the different T cruzi strains. It is concluded that there is a low correlation between the T cruzi strain and the clinical prognosis of Chagas disease, being of greater relevance the host's genetic background and immune response.

Adult↗
PubMed1994

[Evaluation of commercial kits used for Chagas disease diagnosis in blood banks in Chile. II. Routine application].

Aiming to study the applicability and reproducibility of four commercial kits used for the serological detection of Chagas disease (Chagatest-Inst Invest Paraguay, Ortho Chagas, Abbott Chagas (ELISA tests) and Estabilgen Hemo Chagas (indirect hemagglutination test)), a comparative serological study was performed in 256 sera samples coming from a highly endemic area, 249 samples from a low endemic area, 180 reference sera and 2264 samples coming from three blood banks. Specificity of the kits was excellent and sensitivity ranged from 60 to 100%. The indirect hemagglutination test has the lower sensitivity. Some disagreements in the results were observed in the three blood banks, probably due to an unsatisfactory reactive management. We conclude that ELISA tests should be recommended for routine detection of Chagas disease and that for this purpose, a net of laboratories under the direction of a national reference center should exist. This center should assess new commercial products, train technicians and supervise the laboratories.

Blood Banks↗
PubMed1993

[Etiologic diagnosis of pneumonia in HIV-AIDS patients. Usefulness of fibrobronchoscopy].

AIM: to assess the usefulness of fiberoptic bronchoscopy in the etiologic diagnosis of pulmonary infiltrates in patients with AIDS. PATIENTS AND METHODS: in 25 patients with AIDS and pneumonia, 31 fiberoptic bronchoscopies with bronchoalveolar lavage were performed; in 11 occasions, a transbronchial biopsy was also performed. RESULTS: in 24 of the 31 procedures (77%) an etiologic diagnosis was made. There were 7 false negative results; in two of these patients, a repeated examination allowed an etiologic diagnosis. The etiologies of the infiltrates were pyogenic bacteria in 14 cases. Pneumocystis carinii in 9, mycobacteria in 3, cytomegalovirus in one and a lymphocytic interstitial pneumonia in one. In 5 patients, the association of two or more etiological agents was observed. Except one case, all patients had a good response to specific treatments. There were no complications attributed to the bronchoscopic procedure. CONCLUSIONS: fiberoptic bronchoscopy and bronchoalveolar lavage are safe and effective procedures for the etiologic diagnosis of pulmonary infiltrates in patients with AIDS.

AIDS-Related Opportunistic Infections↗
PubMed1993

[The diagnosis of chronic Chagas disease using recombinant antigens of Trypanosoma cruzi].

A panel of eight recombinant Trypanosoma cruzi antigens was used to study the IgG reactivity of sera proceeding from 151 patients with chronic Chagas disease. These samples were reactive against T Cruzi epimastigotes by immunofluorescence and enzyme linked assays. A dot blot assay was performed to evaluate the reactivity against recombinant clones 1, 2, 13, 26, 30, 36, 54 and SAPA (Shed Acute Phase Antigen). Ninety six percent of samples reacted against one or more of these antigens. The most frequently recognized proteins were clones 1, 2, 13, 36 and 26 with 90, 89, 76, 71 and 66 percent of reactivity respectively. Less than 50% of sera reacted against clones 30, 54 and SAPA. Sera from symptomatic patients had a significantly higher frequency of reactivity against clone SAPA than those from asymptomatic patients. This difference suggests that a high reactivity against clone SAPA may be related to acute infections and symptoms. No significant relationship could be established between specific recombinant clone recognitions and particular clinical forms of the disease (cardiological or digestive). It is concluded that these recombinant proteins may become a good tool for the evaluation of the immune response of patients infected with T. cruzi.

Adolescent↗
PubMed1992

Evaluation of an enzyme-linked immunosorbent assay for the diagnosis of Chagas' disease using synthetic peptides.

An enzyme-linked immunosorbent assay (ELISA) has been developed to detect antibodies in human sera to synthetic peptides derived from the repeating amino acid sequence in recombinant Trypanosoma cruzi antigens. Sixty serum samples from patients with chronic Chagas' disease were used to determine the reactivity against the synthetic repeat peptides derived from clones 1, 2, 30, 36, and shed acute phase antigen (SAPA). Ninety-eight percent of the samples had detectable antibodies to one or more of the synthetic peptides at titers greater than 1:100. The percentage of reactive sera increased from 28% with peptide SAPA to 93% with peptide 2. The exposure of patients to T. cruzi was reflected in indirect immunofluorescent antibody titers to fixed epimastigotes. Comparisons between ELISA and immunoradiometric assay data indicated that both tests were of approximately equal sensitivity.

Amino Acid Sequence↗
PubMed1992

Immunodetection of antibodies in sera from symptomatic and asymptomatic Chilean Chagas' disease patients with Trypanosoma cruzi recombinant antigens.

A panel of eight Trypanosoma cruzi antigens produced by recombinant DNA techniques was used to compare the reactivity of IgG specificities in the sera from 45 chronic Chagas' disease patients with different clinical symptoms (cardiac disease, gastrointestinal lesions, and combined syndrome) with those present in the sera from 55 asymptomatic patients in Chile. All of the serum samples were first characterized for antibody to T. cruzi epimastigotes by immunofluorescence assay. All of the Chagas' disease sera were reactive, but none of five healthy controls whose sera were also tested had antibodies against the fixed parasites. A dot-blot assay was then performed to evaluate the serum reactivity against recombinant DNA clones 1, 2, 13, 26, 30, 36, 54, and SAPA (shed acute phase antigen). These recombinant antigens were recognized by a large proportion of the sera collected from the Chilean patients. Ninety-five percent of the serum samples reacted with one or more of the recombinant clones. Analysis of the reactivity with individual fusion proteins showed that 88% of these sera reacted with clones 1 and 2, and 78% reacted with clone 13. Differences in reactivity to clones number 13, 30, and SAPA were observed when symptomatic and asymptomatic patients were compared. These differences in reactivity were statistically significant (P less than 0.01) according to Fisher's exact test.

Adolescent↗
PubMed1992

[Evaluation of commercially available reagents for diagnosis of chagas disease in Chilean blood banks. I. Selection of reagents].

Infection with T cruzi through blood transfusion is an important public health problem in Latin America, including Chile. A study on the sensitivity, specificity and applicability of currently applied methods was performed. Ten commercially available products were tested in a panel of 180 blood specimens against the indirect immunofluorescence antibody test (IFAT) and the immunoperoxidase test (IP). Qualitative concordance ranged from 82 to 98%, sensitivity from 64 to 97% and specificity from 93 to 100%. Some commercial products had a low performance and ELISA is recommended for routine use in Chilean blood banks, given its significantly higher sensitivity and specificity when compared to the hemagglutination test. In addition, experience with the use of the ELISA test is widespread in Chile.

Animals↗
PubMed1992

[Pneumocystis carinii infection. Various aspects on its clinical and laboratory diagnosis].

P carinii is an opportunistic pathogenic agent able to produce severe infection that must be diagnosed promptly. We analyzed 138 samples from 100 patients suspected of having infection by P carinii. The ortho-toluidine blue and the methenamine stains were used to analyze the samples. Infection was demonstrated in 18 patients, 13 adults and 5 children. Underlying disease was AIDS in 7 and other immunosuppressive disorders in the rest. No immunocompetent patient was infected with P carinii. Proper sample collection is important for diagnosis. When bronchoalveolar lavage is not possible, pharyngo-tracheal aspirate in children and sputum sampling after assisted coughing in adults are recommended. At least 2 staining methods and proper controls are advisable.

Adult↗
PubMed1991

Assay for detection of Trypanosoma cruzi antibodies in human sera based on reaction with synthetic peptides.

Synthetic peptides modelled according to the amino acid sequences derived from the repeated domains of five Trypanosoma cruzi antigens were used in an immunoradiometric assay to detect antibodies appearing after natural human infections. An enzyme-linked immunosorbent assay and an indirect immunofluorescence assay performed with a complex antigenic mixture from parasites were used as controls. The results indicate that the synthetic peptides were recognized by a large proportion of serum samples collected from 34 patients with Chagas' disease in Chile and point to their possible use in diagnosis.

Adolescent↗