Parasitological versus molecular diagnosis of strongyloidiasis in serial stool samples: how many?

Strongyloidiasis is usually an asymptomatic disease in immunocompetent patients, caused by Strongyloides stercoralis. However, in immunocompromised patients it can produce a severe clinical profile. Therefore, a correct diagnosis is necessary in these cases and in those chronic asymptomatic patients. The low sensitivity of classical parasitological techniques requires the analysis of multiple serial stool samples. Molecular diagnostic techniques represent an improvement in the detection of the parasite. The objective of this study was to evaluate the minimum number of samples necessary to achieve maximum sensitivity by real-time polymerase chain reaction (PCR). A total of 116 stool samples from 39 patients were analysed by direct microscopic observation, agar culture, Harada-Mori and real-time PCR, in one, two, three and four or more consecutive samples. After two serial samples, 6 out of 39 patients were positive by parasitological and molecular techniques, while 16 of them were real-time PCR positive, and all the patients detected by parasitology were also detected by the molecular technique, reaching 100.00% sensitivity versus 83.00% when analysing a single sample. These data also reflect apparently low specificity (51.52%) and positive predictive value (PPV) (27.27 %) values, due to the high number of cases detected by real-time PCR and not by parasitological techniques. These cases were confirmed as true positives when analysing three, four or more samples from the same patient. In conclusion, the application of molecular techniques decreases the number of serial stool samples necessary to give a diagnosis with the maximum sensitivity.

Análise fitoquímica, potencial antioxidante e toxicidade do extrato bruto etanólico e das frações da espécie Senecio westermaniiDusén frente à Artemia salina / Phytochemical analysis, antioxidant potential and toxicity of crude ethanol extract and fractions of the species Senecio westermanii Dusén against Artemia salina

RESUMO A espécie Senecio westermanii Dusén pertencente à família Asteraceae é planta endêmica e nativa do Brasil encontrada na região da Floresta Atlântica, nos estados do Paraná e São Paulo. O objetivo deste trabalho foi avaliar a composição fitoquímica do extrato bruto etanólico e frações das partes aéreas (folha e caule) de S. westermanii utilizando-se a análise fitoquímica qualitativa e cromatografia líquida de alta eficiência com detector de arranjo de diodos (CLAE-DAD), avaliar in vitro a toxicidade preliminar utilizando Artemia salina e o potencial antioxidante. O estudo fitoquímico qualitativo revelou a presença de alcaloides, flavonoides, iridoides, esteroides/triterpenos, heterosídeos saponínicos e aminogrupos. Através da análise realizada por CLAE-DAD obteve-se o fingerprint característico de cada amostra. No ensaio frente à A. salina houve ausência de toxicidade das amostras, o resultado da DL50 para todas as amostras foi superior a 1000 μg/mL. Todas as amostras apresentaram atividade antioxidante pela redução do complexo fosfomolibdênio, com destaque para a fração clorofórmio que apresentou atividade antioxidante de 92,51% em relação ao padrão rutina. Com relação ao ensaio de redução do radical DPPH•, a fração acetato de etila apresentou IC50 de 26,98 μg/mL. Pelo ensaio do ácido tiobarbitúrico (TBARS) a fração hexano apresentou o melhor índice antioxidante em relação ao padrão rutina. Os resultados obtidos demonstram evidências de que a espécie é fonte potencial de antioxidantes naturais, estimulando assim novos estudos que viabilizam sua utilização no tratamento de patologias associadas aos radicais livres. Além disso, a espécie não apresentou atividade tóxica preliminar, assegurando sua aplicabilidade.

ABSTRACT The species Senecio westermanii Dusén, which belongs to the Asteraceae family, is an endemic and native plant from Brazil. It is found mainly in the Atlantic Forest region, in Paraná and São Paulo states. This study aimed to evaluate the phytochemical composition using a qualitative phytochemical analysis and high performance liquid chromatography with diode array detector (HPLC-DAD), to evaluate preliminary toxicity in vitro using Artemia salina and antioxidant potential. Qualitative phytochemical analysis revealed the presence of alkaloids, flavonoids, iridoids, steroids, triterpenes, saponinic glycosides and aminogroups. The analysis by HPLC-DAD provided the characteristic fingerprint of each sample. In the A. salina assay, the results of LD50 over 1000 μg/mL for all samples did not indicate toxicity of the evaluated extracts. All samples demonstrated antioxidant activity by reducing phosphomolybdenum complex. The highest activity was detected in the chloroform fraction, which presented activity of 92.51% compared to the standard rutin. Regarding the trial to reduce the radical DPPH•, the ethyl acetate fraction showed an IC50 of 26.98 µg/mL. In the thiobarbituric acid assay (TBARS), the hexane fraction showed the highest antioxidant activity compared to the standard rutin. These findings indicate that the Senecio westermanii is a potential source of natural antioxidants, stimulating new studies that enable their use in the treatment of disorders associated with free radicals. In addition, the species did not present toxicity in its evaluation, ensuring its applicability.

Estado de la resistencia a los antibióticos del Vibrio Cholerae en dos hospitales nacionales-Perú: 1991 a 1993 / State of the antibiotic resistance of Vibrio Cholerae in two national hospitals-Peru: from 1991 to 1993

Se estudiaron 53 cepas del Vibrio Cholerae aisladas de pacientes del Hospital Nacional Arzobispo Loayza y 34 procedentes de pacientes pediátricos del Hospital San Bartolomé, con la finalidad de establecer la Concentración Mínima Inhibitoria (CIM) para distintos antibióticos, mediante el método de doble dilución en Agar Muller Hinton. Se encontró que todas las cepas fueron resistentes al Cloramfenicol y a la Lincomicina; además en las cepas aisladas en niños, se halló resistencia a la Gentamicina, Sulfametoxazol y Trimetropin. En el grupo de adultos, la resistencia fue variable, destacando el 11.31% para ampicilina (CIM=0.75 mg/ml), el 13.2% para ácido nalidíxico (CIM=3.5 mg/ml) el 13.20% para penicilina (CIM=3.5 mg/ml) y el 32.1% para amoxicilina (CIM=3.5 mg/ml). En el grupo de cepas aisladas de niños se encontró un 8.82% resistentes a la tetraciclina y a la penicilina (CIM=0.125 y 3.65 UI/ml mg/ml respectivamente), el 5.88% correspondiente a la amoxicilina y ácido nalidíxico (CIM=2.75 mg/ml para ambos) y el 38.23% para ampicilina (CIM=0.75 mg/ml). En relación a los patrones de resistencia a antibiotipos, estos varían de R:3 a R:9; siendo los más frecuentes el R:5 (Cloramfenicol, Clindamicina, Lincomicina, Sulfametoxazol y Trimetropin) en adultos y R:4 (Clindamicina, Lincomicina, Gentamicina y Trimetropin) en niños. Estos resultados sugieren que el Vibrio Cholerae actuaría como posible reservorio de plásmidos de resistencia a antibióticos, lo que complicaría el tratamiento de enfermedades y al mismo tiempo la erradicación del Vibrio Cholerae de nuestro medio.

Fifty three strains of the Vibrio Cholerae were studied, isolated from patients from the National Hospital Arzobispo Loayza and 34 strains coming from pediatric patients from the Bartolome Hospital, with the goal of establishing the Minimun Inhibitory Concentration (MIC) for different antibiotics, by means of the method of double dilution in Agar Muller Hinton. It was found that all the strains were resistant to Cloramfenicol and Lincomicin; besides, at the isolated strains from children, was found a resistance to Gentamicin, Sulfametoxazol and Trimetropin. At the adult group the resistance was variable, being remarkable the 11.32% from ampicilina (MIC=0.75 ug/ml) the 13% for nalidixic acid (MIC=3.5 ug/ml). At the group of isolated strains from children, it was found an 8.82% resistant to tetraciclina and penicillin (MIC=0.152 and 3.65 UL/ml ug/ml respectively) the 5.88% belong to amoxilin and nalidix acid (MIC=2.75 ug/ml for both) and 38.23% for ampicilina (MIC=0.75 ug/ml). In relation to the patterns of resistance to anti-biotic types, these change from r:3 to r:9 being the more frequent ones the R:5 (Cloramfenicol, Clindamicina, Lincomicin, Sulfametoxazol and Trimetropin) in adults and R:4 (Clindamicina, Lincomicin, Gentamicin and Trimetropin) in children. These results suggest that the Vibrio Cholerae will act as a possible reservoir of plasmids of resistance to anti-biotics, and that would complicate the treatment of the illness, and at the same time, the ending of Vibrio Cholerae in our environment.

Na,K-ATPase activity in red blood cells from patients with Chediak-Higashi syndrome.

Na,K-ATPase activity of red blood cells from Chediak-Higashi syndrome (CHS) patients and relatives (gene heterozygous) was determined and compared to that of control, healthy, individuals. The enzyme activity was found to be strongly diminished in the CHS patients and slightly lower in their relatives. This reduced activity was due to a lower turnover number of the Na, K-ATPase as well as a decreased number of pumps. The reduced enzyme activity observed in these patients could be the result of an abnormal cell membrane fluidity, and the lowered number of Na, K-pumps could be explained as a consequence of an altered or deficient cell machinery caused by the CHS gene.

Una alternativa para la detección de Vibio cholerae en aguas / An alternative for the detection of Vibio cholerae in waters

Se ha desarrollado un método que podría ser usado como alternativa en la detección indirecta de Vibrio cholerae en aguas, mediante la utilización de vibriófagos aislados de aguas marinas someras, relacionadas con desembocaduras de dos ríos y con colector de aguas servidas. Para tal efecto, se han tenido en cuenta las características de crecimiento de este microorganismo, así como la especificidad de sus fagos. Se ha conseguido calificar y cuantificar la presencia del vibrión colérico en los lugares muestreados.

We have developed a method that could be used as an alternative for indirect detection of Vibrio cholerae from water through the use of vibriophages isolated from shallow marine water relating to two river mouths and to a sewage pipe- for this, we have considered the growing characteristics of this microorganism as well as its phage specifity. We have qualified and quantified the presence of the choleric vibrion in the sampled places.

[Human immunodeficiency virus infection in the Spanish population. Review of the literature]. / Infección por el virus de la inmunodeficiencia humana en la población española. Revisión de la literatura.

The epidemiological aspects of the infection by the human immunodeficiency virus, HIV, in the Spanish population as described in the literature are reviewed. From the published studies is evident that HIV infection is limited to the population risk groups established in the epidemiological world reports, heroin addicts, homosexuals, heterosexuals with sexual contacts with infected persons and blood and hemoderivates receptors. The observed frequencies in these risk groups are similar to that described in other countries.

[Human immunodeficiency virus (HIV) infection in País Vasco, Spain]. / Infección por el virus de inmunodeficiencia humana (VIH) en el País Vasco, España.

An analysis of the epidemiology of the human immunodeficiency virus infection in the Basque Country, Spain is presented. The infection by this virus is mainly detected in intravenous drug addicts. In homosexuals the seropositivity frequency detected was moderate, and infection was also detected in sexual partners of drug addicts.

[Diseases in heroin addiction]. / Patologías médicas en la adicción a la heroína.

An analysis of the medical pathologies seen in heroin addicts heroin addicts hospitalized in the Provincia de Vizcaya, Basque Country, Spain, from 1982 to 1987, and its possible association to the infection by the human immunodeficiency virus, HIV, is presented. An increasing of hospitalized addicts per year was observed, being infectious diseases the main pathology detected. HIV infection was present in more than 90% of individuals with non opportunistic acquired infections. Significant annual increase in the number of tuberculosis and candida infections was present. The possible association of HIV infection with increased susceptibility to commonly acquired non opportunistic infections is discussed.

[Epidemiologic aspects of acquired immunodeficiency syndrome (AIDS). Review]. / Aspectos epidemiológicos del sindrome de immunodeficiencia adquirida (SIDA). Revisión.

A revision of the World current epidemiological aspects of the acquired immunodeficiency syndrome is presented. It was observed that near 70% of all the world cases occurred in the United States of America and 12% in Europe. There is an increase tendency of the number of cases. However, the analysis of the inter-annual percentage increase showed a tendency to stabilization in Europe and in the United States of North America.

Manejo del vólvulo de sigmoides: 1981-1986 / Management of sigmoid volvulus: 1981-1986

El vólvulo de sigmoide es la torsión del intestino sobre si mismo en relación con su eje longitudinal, con obstrucción parcial o completa de la luz intestinal en ambos extremos del segmento, produciendose oclusión en asa cerrada con alteraciones variables de la circulación. En el departamento de Cochabamba, es la causa más frecuente de obstrucción intestinal baja.

Leucemias en Venezuela asociadas al virus humano de leucemia/linfoma tipo I (HTLV-1) / Human T cells leukaemia/limphoma virus type I (HTLV-1) associated leukaemias in Venezuela

Se describen 5 casos de leucemias positivas para el virus humano de leucemia/linfoma tipo I (HTLV-1). Tres de los pacientes eran nativos de una región de alta endemicidad para este virus, (Estado Zulia, Venezuela) y padecían de leucemia aguda linfoblástica o indiferenciada. Los otros dos pacientes eran originarios de una zona de baja endemicidad (Caracas), con leucemia linfocítica crónica y promielocítica aguda. El estudio de los marcadores de superficie celular no mostró evidencia de un fenotipo de leucemia de célula T del adulto en ningún caso y sugiere que la infección por HTLV-1 puede estar asociada con desórdenes proliferativos no T. Alternativamente HTLV-1 puede ser el agente etiológico de una variedad de formas de leucemias, específicamente en las áreas de alta endemicidad en el Caribe

Antibodies to acquired immune deficiency syndrome (AIDS)-associated virus (HTLV-III/LAV) in Venezuelan populations.

Serum samples from 850 individuals from Venezuela were tested for the presence of antibodies to HTLV-III/LAV virus, the probable etiological agent of acquired immune deficiency syndrome (AIDS). At the time of the study, none of the individuals tested had symptoms indicative of AIDS or related disorders. Viral antibodies were assayed by indirect immunofluorescence (IF) assay, using a chronically infected, HTLV-III/LAV producer cell line CEM/LAV-NIT established in our laboratory. Twenty individuals (2.5%), 8 of them (40%) female, were seropositive by IF and by confirmatory Western blotting and radioimmunoprecipitation assays. The seropositivity rate ranged from 2.4% (11 of 465) in the general healthy population, 4% (2 of 50) among patients with Chagas' disease, and up to 29.2% (7 of 24) among patients with acute malaria infection. The titers of HTLV-III/LAV antibodies ranged from 1:40 to 1:640. In addition, 2 of 36 patients with hemophilia A (5.5%) also had antibodies to HTLV-III/LAV. Two of 7 patients with acute malaria had specific antibodies both to HTLV-III/LAV and HTLV-I, as determined by IF and Western blotting. None of over 169 randomly chosen, healthy blood donors from seven major Venezuelan cities, as well as none of 99 patients with leukemia/lymphoma, had antibodies to HTLV-III/LAV. The presence of specific antibodies among various Venezuelan populations indicates that HTLV-III/LAV, or a closely related cross-reactive virus, is indigenous in Latin American subjects as was previously indicated for tropical populations of central Africa. Isolation and characterization of this virus will help to understand the origin and etiology of AIDS.

Antibodies to HTLV-III/LAV among aboriginal Amazonian Indians in Venezuela.

Serum samples from 224 aboriginal Amazonian Indians were tested for antibodies to HTLV-III/LAV by an indirect immunofluorescence (IF) assay. 9 individuals (4%), 5 of them female, were seropositive by IF and by confirmatory western blotting and radioimmunoprecipitation tests. 3 of the positive sera were collected in 1968. HTLV-III/LAV seropositivity rates varied among the ethnic groups and ranged from 13.3% among the Pemon Indians to 3.3% among the Yanoama tribe. The titres of HTLV-III/LAV antibodies ranged from 1/40 to 1/320. All individuals tested were apparently healthy at the time of the study. None of 211 randomly chosen, healthy blood donors from Venezuelan cities had antibodies to HTLV-III/LAV. The prevalence of specific antibodies among Amazonian Indians suggests the HTLV-III/LAV or a closely related cross-reactive virus may be endemic in this area. The findings also indicate that this virus is indigenous in non-negroid Latin American and negroid tropical populations.

Chédiak-Higashi syndrome in a Venezuelan black child.

One case of Chédiak-Higashi syndrome (CHS) in a black male child, born to a consanguineous couple from a rural village in the State of Falcón, is described. At birth the child had marked skin depigmentation and ash-gray hair. A few months later he developed an almost normal black skin color. The diagnosis of CHS was established by the presence of large peroxidase-positive granules in his leukocytes. Neutrophils showed decreased chemotaxis and lack of digestive capacity against Candida albicans. Unusual features included extreme rarity of CHS in blacks, progressive repigmentation of the skin, and an early benign evolution. A high consanguinity index in the village from which this patient originated raised the possibility of the presence of a new cluster of this disease in Venezuela.

Natural antibodies to human T-cell leukemia/lymphoma virus in healthy Venezuelan populations.

Serum samples of 769 healthy Venezuelan donors were assayed for natural antibodies to HTLV-I by the ELISA technique. Specific HTLV-I antibody prevalence was 6.8% but varied from 1% in Caracas to 13.7% in the Amazonas region and the State of Zulia. Adults infected with Trypanosoma cruzi had the highest HTLV-I antibody prevalence of 15%. Areas of high antibody prevalence were correlated most strongly with the presence of arthropod-borne diseases and to a lesser extent with socio-economic factors. Genetic factors were not correlated with antibody prevalence. Antibodies were seen in children as young as 3 years of age in the most endemic areas. Antibody titers increased with age, suggesting continuous exposure to the virus. The data provide clues for elucidation of the geographic variation in HTLV-I antibody prevalence seen Venezuela and other HTLV-I endemic areas. In addition, they further confirm the Caribbean region as being endemic for HTLV-I and extend this region to inland areas of South America.

Functions of neutrophils in endemic Chediak-Higashi syndrome.

Endemic Chediak-Higashi Syndrome occurs in a restricted geographic area (Pregonero, State of Táchira, Venezuela). Neutrophils from these patients were unable to digest Candida albicans in vitro, but showed normal or increased metabolic activities. This finding supports the view that the endemic syndrome is bona fide Chediak-Higashi Syndrome.

Human T cell leukemia virus type I and Epstein-Barr virus antibodies in homosexuals and hemophilia patients from Venezuela.

The presence of serum antibodies to human T-cell leukemia/lymphoma Virus-I (HTLV-I) and to Epstein-Barr Virus (EBV) was studied in homosexuals and hemophilia patients from the State of Zulia (Venezuela), a highly endemic area for HTLV-I infection. No serum antibodies to HTLV-I were detected despite the presence of alterations in peripheral blood lymphocyte subsets. Anti-EBV antibodies in homosexuals were indicative of persistent infection with this virus. Our results suggest that HTLV-I does not play a role in the immunological alterations in homosexuals or hemophiliacs in an area of high prevalence for this virus.