Polimorfismos de los genes CCR5, IL-6, IFN-γ e IL-10 en pacientes cubanos con VIH/sida / Polymorphisms of CCR5, IL-6, IFN-γ and IL-10 genes in Cuban HIV/AIDS patients

Introducción Los estudios genéticos han demostrado asociaciones de varios polimorfismos de un solo nucleótido (SNP) con diferentes tasas de progresión y variación en la susceptibilidad a la infección por VIH. Este estudio tuvo como objetivo estimar la frecuencia de los polimorfismos ccr5Δ32, IL-6-174G/C, IFN-γ+874T/A e IL-10-1082A/G en pacientes cubanos infectados por VIH y un grupo de parejas serodiscordantes para evaluar su influencia sobre el riesgo y la progresión de la enfermedad. Métodos Se realizó un estudio transversal en 120 sujetos atendidos en el Instituto de Medicina Tropical «Pedro Kour» (IPK) y el Hospital Hermanos Ameijeiras entre junio de 2018 y diciembre de 2019. La amplificación de los fragmentos de los genes ccr5, IL-6, IFN-γ e IL-10 se realizó mediante reacción en cadena de la polimerasa seguida por el análisis del polimorfismo de fragmentos de restricción utilizando la enzima NlaIII para la IL-6. El sistema de mutación refractario a la amplificación por PCR se utilizó en el caso de los genes IFN-γ e IL-10. Resultados Las distribuciones alélicas y genotípicas de los genes ccr5, IL-6, IFN-γ e IL-10 no difirieron significativamente entre los dos grupos. Los recuentos celulares y los valores de carga viral en plasma no difirieron significativamente entre los genotipos de los genes ccr5, IL-6, IFN-γ e IL-10. Solo el genotipo IL-6 GC se asoció con valores más altos de carga viral. La combinación de alelos de los cuatro SNP considerados mostró un aumento muy significativo del riesgo de infección por VIH para uno de ellos, pero con una frecuencia muy baja (<1%). Conclusión Este estudio contribuye a evaluar la frecuencia de estos polimorfismos y su influencia en los biomarcadores de la progresión de la infección por VIH en la población cubana con infección por el VIH. (AU)

Introduction Genetic studies have shown associations of several single nucleotide polymorphisms (SNP) with different rates of progression and variation in susceptibility to HIV infection. This study aimed to estimate the frequency of ccr5Δ32, IL-6-174G/C, IFN-γ+874T/A and IL-10-1082A/G polymorphisms in Cuban HIV-infected patients and a group of sero-discordant couples to assess their influence on risk and disease progression. Methods A cross-sectional study was carried out on 120 subjects registered at the Institute of Tropical Medicine «Pedro Kour» (IPK) and the Ameijeiras Hospital from June 2018 until December 2019. The amplification of fragments of the ccr5, IL-6, IFN-γ and IL-10 genes was performed by polymerase chain reaction followed by identification of polymorphisms using the restriction fragment length polymorphism analysis for IL-6 with the restriction enzymes Nla III. Amplification refractory mutation system was used for IFN-γ and IL-10 genes. Results The allelic and genotypic distributions of the genes ccr5, IL-6, IFN-γ and IL-10 did not differ significantly between the two groups. Cell counts and plasma viral load values did not differ significantly between genotypes of the ccr5, IL-6, IFN-γ and IL-10 genes. Only the IL-6 GC genotype was associated with higher viral load values. The combination of alleles of the four considered SNPs showed a highly significant increase in the risk of HIV infection for one of them, but with a very low frequency (<1%). Conclusion This study contributes to evaluating the frequency of these polymorphisms and their influence on biomarkers of the progression of HIV infection in the Cuban HIV-population. (AU)

Polimorfismos de los genes CCR5, IL-6, IFN-γ e IL-10 en pacientes cubanos con VIH/sida / Polymorphisms of CCR5, IL-6, IFN-γ and IL-10 genes in Cuban HIV/AIDS patients

Introducción Los estudios genéticos han demostrado asociaciones de varios polimorfismos de un solo nucleótido (SNP) con diferentes tasas de progresión y variación en la susceptibilidad a la infección por VIH. Este estudio tuvo como objetivo estimar la frecuencia de los polimorfismos ccr5Δ32, IL-6-174G/C, IFN-γ+874T/A e IL-10-1082A/G en pacientes cubanos infectados por VIH y un grupo de parejas serodiscordantes para evaluar su influencia sobre el riesgo y la progresión de la enfermedad. Métodos Se realizó un estudio transversal en 120 sujetos atendidos en el Instituto de Medicina Tropical «Pedro Kour» (IPK) y el Hospital Hermanos Ameijeiras entre junio de 2018 y diciembre de 2019. La amplificación de los fragmentos de los genes ccr5, IL-6, IFN-γ e IL-10 se realizó mediante reacción en cadena de la polimerasa seguida por el análisis del polimorfismo de fragmentos de restricción utilizando la enzima NlaIII para la IL-6. El sistema de mutación refractario a la amplificación por PCR se utilizó en el caso de los genes IFN-γ e IL-10. Resultados Las distribuciones alélicas y genotípicas de los genes ccr5, IL-6, IFN-γ e IL-10 no difirieron significativamente entre los dos grupos. Los recuentos celulares y los valores de carga viral en plasma no difirieron significativamente entre los genotipos de los genes ccr5, IL-6, IFN-γ e IL-10. Solo el genotipo IL-6 GC se asoció con valores más altos de carga viral. La combinación de alelos de los cuatro SNP considerados mostró un aumento muy significativo del riesgo de infección por VIH para uno de ellos, pero con una frecuencia muy baja (<1%). Conclusión Este estudio contribuye a evaluar la frecuencia de estos polimorfismos y su influencia en los biomarcadores de la progresión de la infección por VIH en la población cubana con infección por el VIH. (AU)

Introduction Genetic studies have shown associations of several single nucleotide polymorphisms (SNP) with different rates of progression and variation in susceptibility to HIV infection. This study aimed to estimate the frequency of ccr5Δ32, IL-6-174G/C, IFN-γ+874T/A and IL-10-1082A/G polymorphisms in Cuban HIV-infected patients and a group of sero-discordant couples to assess their influence on risk and disease progression. Methods A cross-sectional study was carried out on 120 subjects registered at the Institute of Tropical Medicine «Pedro Kour» (IPK) and the Ameijeiras Hospital from June 2018 until December 2019. The amplification of fragments of the ccr5, IL-6, IFN-γ and IL-10 genes was performed by polymerase chain reaction followed by identification of polymorphisms using the restriction fragment length polymorphism analysis for IL-6 with the restriction enzymes Nla III. Amplification refractory mutation system was used for IFN-γ and IL-10 genes. Results The allelic and genotypic distributions of the genes ccr5, IL-6, IFN-γ and IL-10 did not differ significantly between the two groups. Cell counts and plasma viral load values did not differ significantly between genotypes of the ccr5, IL-6, IFN-γ and IL-10 genes. Only the IL-6 GC genotype was associated with higher viral load values. The combination of alleles of the four considered SNPs showed a highly significant increase in the risk of HIV infection for one of them, but with a very low frequency (<1%). Conclusion This study contributes to evaluating the frequency of these polymorphisms and their influence on biomarkers of the progression of HIV infection in the Cuban HIV-population. (AU)

Polymorphisms of CCR5, IL-6, IFN-γ and IL-10 genes in Cuban HIV/AIDS patients.

INTRODUCTION: Genetic studies have shown associations of several single nucleotide polymorphisms (SNP) with different rates of progression and variation in susceptibility to HIV infection. This study aimed to estimate the frequency of ccr5Δ32, IL-6-174G/C, IFN-γ+874T/A and IL-10-1082A/G polymorphisms in Cuban HIV-infected patients and a group of sero-discordant couples to assess their influence on risk and disease progression. METHODS: A cross-sectional study was carried out on 120 subjects registered at the Institute of Tropical Medicine «Pedro Kour¼ (IPK) and the Ameijeiras Hospital from June 2018 until December 2019. The amplification of fragments of the ccr5, IL-6, IFN-γ and IL-10 genes was performed by polymerase chain reaction followed by identification of polymorphisms using the restriction fragment length polymorphism analysis for IL-6 with the restriction enzymes Nla III. Amplification Refractory Mutation System was used for IFN-γ and IL-10 genes. RESULTS: The allelic and genotypic distributions of the genes ccr5, IL-6, IFN-γ and IL-10 did not differ significantly between the two groups. Cell counts and plasma viral load values did not differ significantly between genotypes of the ccr5, IL-6, IFN-γ and IL-10 genes. Only the IL-6 GC genotype was associated with higher viral load values. The combination of alleles of the four considered SNPs showed a highly significant increase in the risk of HIV infection for one of them, but with a very low frequency (<1%). CONCLUSION: This study contributes to evaluating the frequency of these polymorphisms and their influence on biomarkers of the progression of HIV infection in the Cuban HIV-population.

First record of the Lesser Snouted Treefrog Scinax nasicus (Cope, 1862) in Brazilian coast and new species records for the state of Rio Grande do Sul / Primeiro registro da perereca-de-banheiro Scinax nasicus (Cope, 1862) na costa do Brasil e novos registros para o estado do Rio Grande do Sul

Abstract Herein, we provide new occurrence records of Scinax nasicus (Cope, 1862) for the state of Rio Grande do Sul, Southern Brazil. All new records here provide are located on Southern half of the state. Besides this, we provide the first record for species in Brazilian coastal zone. Those records improve considerably our knowledge regarding species distribution in Southern Brazil.

Resumo Aqui, nós fornecemos novos registros de ocorrência de Scinax nasicus (Cope, 1862) para o estado do Rio Grande do Sul, Sul do Brasil. Todos os novos registros aqui fornecidos estão localizados na metade sul do estado. Além disso, nós fornecemos o primeiro registro para a espécie na zona costeira brasileira. Esses registros melhoram consideravelmente o nosso conhecimento sobre a distribuição da espécie no Sul do Brasil.

First record of the Lesser Snouted Treefrog Scinax nasicus (Cope, 1862) in Brazilian coast and new species records for the state of Rio Grande do Sul.

Herein, we provide new occurrence records of Scinax nasicus (Cope, 1862) for the state of Rio Grande do Sul, Southern Brazil. All new records here provide are located on Southern half of the state. Besides this, we provide the first record for species in Brazilian coastal zone. Those records improve considerably our knowledge regarding species distribution in Southern Brazil.

Incidence of primary and second cancers in renal transplant recipients: a multicenter cohort study.

Limited data exist about cancer prognosis and the development of second cancers in renal transplant recipients. In a retrospective cohort study on 3537 patients incidence rates of the first and, if any, of a second cancer, and standardized incidence ratios [SIR (95% CI)] were computed. Two hundred and sixty-three (7.5%) patients developed a NMSC, and 253 (7.2%) another type of cancer after a median follow-up of 6.5 and 9.0 years, respectively. A statistically significant excess risk, if compared to an age- and sex-matched reference general population, was observed for Kaposi sarcoma and NMSC, followed by non-Hodgkin lymphoma and carcinoma of cervix uteri; a small number of unusual cancers such as tumors of the salivary glands, small intestine and thyroid also were detected at a level worthy of additional scrutiny. Ten-year survival rate of all noncutaneous cancers was 71.3%, with lower rates for lung carcinoma and non-Hodgkin lymphoma (0% and 41.7%, respectively). Patients with NMSC had an increased risk of developing a second NMSC [SIR 8.3 (7.0-10.0)], and patients with a primary noncutaneous cancer had increased risk of developing a second noncutaneous cancer [SIR 1.8 (1.2-2.8)], if compared to the whole cohort. Our study underscore that the high risk of primary and second cancer in renal transplant recipients, including unusual cancers.

[Preadolescents' nutritional habits: a survey in the secondary schools in Brianza]. / Le abitudini alimentari dei preadolescenti: indagine nelle scuole medie della Brianza.

AIM: This study was aimed at assessing eating habits among preadolescents living in Brianza, with a special focus on snacking' and breakfast' habits. METHODS: The research was carried out in 12 post-primary schools in Brianza and included 802 students attending 43 different classes (49.3% males and 50.7% females; mean age:12.6 years). The research tool was an ad hoc questionnaire administered to the preadolescents attending school on the day of the research. Data were analyzed using program Epi Info 6. RESULTS: Many students (78.3%) had breakfast in the morning of the questionnaire's administration. Among those perceiving themselves as overweight, not having breakfast is more common than among others. This information is the opposite of what preadolescents say when inquired about their habit to have ''a good breakfast'': only 36.8% of the responders declare that it happens on a daily basis. As to snacking, 18.7% of the students declare to be used to having multiple snacking during the day; during school recess on the day of the survey, 39.9% of respondents declared that they had eaten various industrial snacks, while 27.4% had cakes or fresh bread, 1.2% both of them and 0.6% fruit. On a daily basis, 59.5% report to eat various and different fruits and 62.9% report the same when inquired about vegetables. CONCLUSION: Considering the importance of a correct nutrition during developmental age and the importance of the phase of adolescence in establishing permanent nutritional habits that will last life-long, it is thus an educational priority to help preadolescents to choose the best foods for their health and well being.

Antibodies from patients with dengue viral infection mediate cellular cytotoxicity.

Acute and late convalescent sera (collected at day 5 of disease onset and 1 year later) from dengue fever (DF) and dengue hemorrhagic fever/dengue shock syndrome (DHF/DSS) laboratory confirmed cases, were tested for antibody-dependent cell-mediated cytotoxicity (ADCC) activity using dengue 1 (DENV-1) or dengue 2 (DENV-2) infected cells as target. All patients experienced their first dengue virus (DENV) infection 20 years before. ADCC activity was detected in acute sera from DHF/DSS but not in sera from DF patients. However, 1 year after illness, ADCC activity was observed in all cases. This preliminary report represents one of the few studies of ADCC in dengue patients and suggests that ADCC could be implicated in dengue pathogenesis.

Mobilization of a hobo-related sequence in the genome of Drosophila simulans.

The hobo transposable element can occur under three forms in the Drosophila genome: as a complete element (also called canonical), as internally deleted copies, or as hobo-related sequences (relics). Some evidence indicated that canonical elements and internally deleted copies are recent acquisitions of Drosophila genomes, while the "relics" are old components, normally degenerated and immobile. Here we present the characterization of a hobo-related sequence, found in the genome of a hypermutable strain of D. simulans, which insertion into the white locus raised a de novo white mutation. It is a shorter hobo related element presenting, overall, roughly 18% of divergence at the DNA level from the canonical hobo, with many indels that make clear this element is defective. However, its ITRs and flanking regions are extremely conserved. This is the first hobo "relic" showed to be mobilizable. We suggest, and point up some evidences, toward the idea that this sequence could have been mobilized by the canonical element. The presence of a similar "relic" element in D. sechellia allows us to suggest that these elements have been maintained mobilizable since the time of divergence between these species.

[Pre-adolescents and sport: results of a health survey in some secondary schools of Brianza]. / I preadolescenti e lo sport: risultati di un'indagine in alcune scuole medie dell'area brianzola.

This study is aimed at assessing time dedicated to, motivation and involvement in physical activity on behalf of pre-adolescents. 802 students (49.3% boys and 50.7% girls; mean age: 12.6) attending 43 classes of 11 Brianza's post-elementary schools. A questionnaire was prepared and administered to the involved sample. Classes took part in this study through randomized selection and data were analyzed using program Epi Info 6. Most of the respondents (60.1%) stay involved in sport because "it makes me feel good" and 32.4% because sport is "enjoyable and entertaining". Pre-adolescents chose the sport in which they become engaged on the basis of personal inclination (40%), in order to share experiences and their free time with friends (15.4%); 13.7% declare to prefer sport to be practiced in team. (13.7%). The most practiced sport are: football (51.3% boys, 4.5% girls), volley (3.9%-36.4%), dance (0.8%-25.2%) e swimming (9.4%-14%). 80.8% in the sample practice physical activity in settings outside schools and, among these, 51.7% dedicate three or more hours weekly to extra-school activities. There are significant differences (p < 0.05) between males' and females' engagement in physical activity (mean value: 4.1 hours; males: 4.6 h, females: 3.5 h). Among pre-teens, 18% declare to be physically inactive at all; 15.7% do not take part in sport activities because afraid to be bullied or shamed by peers. Research show that males are significantly more involved in physical activity than females.

Multiplex polymerase chain reaction amplification and differentiation of Entamoeba histolytica and Entamoeba dispar DNA from stool samples.

Due to the clinical importance of differentiating the two species of the Entamoeba histolytica/Entamoeba dispar complex, we developed a multiplex polymerase chain reaction (PCR) method that overcomes time-consuming and laborious procedures. We report here a DNA extraction protocol using non-fixed stool samples that avoid long lysis-incubation periods through the combined use of zirconium beads and a lysis-supporting buffer. We characterized 49 of 52 stool specimens from Cuban patients with amoebiosis. Among them, 36 (75.5%) were infected only with E. dispar (the nonpathogenic species), while 13 (24.5%) displayed a mixed infection with both E. dispar and E. histolytica. The multiplex PCR protocol showed a specificity of 1.00 and a sensitivity of 0.94. Furthermore, the entire procedure can be performed in one day. This approach is therefore reliable and applicable in the field for epidemiologic studies.

Specific cellular and humoral immune response in Balb/c mice immunised with an expression genomic library of Trypanosoma cruzi.

An expression genomic library of Trypanosoma cruzi (T. cruzi) constructed using pcDNA3 plasmid was used for the immunisation (25 micrograms) of Balb/c mice. Expression of T. cruzi antigens in the muscle of inoculated mice was detected by indirect immunofluorescence 7 days after immunisation. Specific IgG antibodies were significatively increased (P < 0.05) in animals that were reimmunized with 50 micrograms of the genomic library. An antigen specific lymphoproliferative response was detected in one animal of the group inoculated with one dose of the library.

[Identification of mosquitoes' human food source by using the co-agglutination technique]. / Identificación de una fuente humana de alimentación de mosquitos mediante la técnica de coaglutinación.

The utilization of a coagglutination technique for the identification of a human source for feeding mosquitoes is described. The dilution of ingested blood samples in filter paper was performed in 2 mL of a sodium chloride solution at 0.85%. It was used a suspension of sensibilized Staphylococcus aureus with rabbit's serum, human plasmatic anti-proteins, and human anti-IgG rabbit's serum discriminated well between human and non human blood. No agglutination was observed with the negative control. This technique proved to be sensitive to identify 100% of the human blood samples taken to the paper 24 hours after the mosquitoes completed their feeding at a temperature of 26 to 28 degrees C. Among mosquitoes fed and collected in the fields the test had a satisfactory result. Therefore, it may be used in routine work in the fields. The results showed the sensitivity and specificity of this method for identifying human blood ingested by mosquitoes.

Toxoplasma gondii antigenuria in patients with acquired immune deficiency syndrome.

A longitudinal study was performed with sera and urine of patients with acquired immune deficiency syndrome (AIDS), taken before, during and after clinically Toxoplasma infection. The tested patients were followed for an average of two years. The titres of the specific IgG and IgM antibodies were measured by an indirect fluorescent antibody test (IFAT), and the appearance of circulating antigens of T. gondii was determined in 36 urine samples of 13 patients with neurotoxoplasmosis by means of the coagglutination test. The presence of T. gondii antigens in the urine of AIDS patients by this test was correlated with the immunoblot technique, with clinical symptoms and also with pathological findings. Our results indicate that the detection of T. gondii antigens in the urine of AIDS patients can be regarded as a rapid and efficient method for the diagnosis of acute toxoplasmosis.

Usefulness of the detection of Toxoplasma gondii antigens in AIDS patients.

Toxoplasmic encephalitis (TE) is a mayor cause of central nervous system infection in patients with acquired immunodeficiency syndrome (AIDS). Toxoplasma antibodies were detected in 56 of 79 patients with AIDS (71%), in the present study. Fourteen out of 57 seropositive patients developed TF (25%) and had Toxoplasma gondii antigen detected in their urine. For this, most of them received an effective therapy, with the subsequent disappearance of the symptoms and discontinuity of excretion of the T. gondii antigens. Our results suggest that the monitoring of T. gondii antigen in the urine of AIDS patients may be useful to decide on the proper time for therapy, as well as to avoid the beginning of neurologic signs in these patients.

Inhibition of human monocyte function by prophylactic doses of chloroquine.

The effect of prophylactic doses of chloroquine on the phagocytic function of human monocytes was studied in young healthy male volunteers. They received placebo, 300, or 600 mg of chloroquine base/week for six weeks. In each subject, the phagocytic function was tested before and at the end of the chloroquine intake period. In the 600-mg chloroquine group, it was also tested six weeks after receiving the last dose. Chloroquine at both doses inhibited the phagocytosis of IgG-coated sheep red blood cells and of zymosan particles. The effect was more pronounced with the 600 mg dose of chloroquine. The phagocytic activity returned to normal values six weeks after the end of treatment.

Identification of Dirofilaria immitis-infected dogs with a coagglutination assay.

The presence of Dirofilaria immitis excretory-secretory (ES) products was detected in the urine of infected dogs using a coagglutination assay. Urine samples from 30 naturally infected dogs were positive. Seventeen of them were microfilaremic, whereas 13 had become amicrofilaremic after receiving 2 courses of diethylcarbamazine. Urine samples from 20 dogs infected with other parasites, Dipetalonema reconditum (7), Toxocara canis (5), and Ancylostoma caninum (8), and urine samples from 20 healthy dogs were negative. The assay detected 200 ng/ml or more of ES products. This assay is simple, easy to perform with minimum training, and requires no equipment. Therefore it should be useful to detect canine filariasis under field conditions.