Cytogenetic changes in oral mucosa cells from individuals submitted to oral human immunodeficiency virus pre-exposure prophylaxis use.

OBJECTIVE: The objective of this study was to evaluate cytogenetic changes in individuals submitted to oral human immunodeficiency virus pre-exposure prophylaxis use through the micronucleus test in oral mucosa. METHODS: This study consisted of 37 individuals, of whom 17 comprised the pre-exposure prophylaxis group and 20 comprised the control group. A total of 2,000 cells per slide were analyzed for the determination of micronuclei, binucleation, nuclear buds, and cytotoxicity parameters: pyknosis, karyolysis, and karyorrhexis (KR), in a double-blind manner. The repair index was also evaluated in this setting. RESULTS: In the mutagenicity parameters, the pre-exposure prophylaxis group showed increased frequencies of micronuclei (p=0.0001), binucleation (p=0.001), and nuclear buds (p=0.07). Regarding the cytotoxicity parameters, there was an increase with a statistical difference (p≤0.05) in the karyorrhexis frequency (p=0.001). Additionally, the repair system efficiency decreased in the pre-exposure prophylaxis group. CONCLUSION: These results indicate that individuals undergoing pre-exposure prophylaxis use have geno- and cytotoxicity in oral mucosal cells.

Cytogenetic changes in oral mucosal cells of human immunodeficiency virus-infected children and adolescents undergoing antiretroviral treatment.

OBJECTIVE: The objective of this study was to evaluate possible cytogenetic changes in children and adolescents with human immunodeficiency virus on antiretroviral therapy, through the micronucleus test in oral mucosa. METHODS: This was a prospective study consisted of 40 individuals, of whom 21 comprised the human immunodeficiency virus group and 19 comprised the control group. Children and adolescents with human immunodeficiency virus were enrolled. The inclusion criteria were <18 years old and consent in participating in the study. The exclusion criteria were the presence of numerous systemic comorbidities, oral lesions, the habit of smoking, alcohol consumption, and X-rays or CT scans taken within 15 days prior to sample collection. A gentle scraping was performed on the inner portion of the jugal mucosa on both sides. A total of 2,000 cells per slide were analyzed for the determination of mutagenicity parameters as follows: micronuclei, binucleation, and nuclear buds. For measuring cytotoxicity, the following metanuclear changes were evaluated: pyknosis, karyolysis, and karyorrhexis, in a double-blind manner. The repair index was also evaluated in this setting. RESULTS: The human immunodeficiency virus group showed high frequencies of micronuclei (p=0.05), binucleated cells (p=0.001), and nuclear buds (p=0.03). In the cytotoxicity parameters, represented by the cell death phases, there was an increase with statistical difference (p≤0.05) in the karyorrhexis frequency (p=0.05). Additionally, repair index was decreased in the human immunodeficiency virus group. CONCLUSION: These results indicate that human immunodeficiency virus -infected individuals undergoing antiretroviral therapy have cytogenetic changes in oral mucosal cells.

Cytogenetic changes in oral mucosal cells of human immunodeficiency virus-infected children and adolescents undergoing antiretroviral treatment

SUMMARY OBJECTIVE: The objective of this study was to evaluate possible cytogenetic changes in children and adolescents with human immunodeficiency virus on antiretroviral therapy, through the micronucleus test in oral mucosa. METHODS: This was a prospective study consisted of 40 individuals, of whom 21 comprised the human immunodeficiency virus group and 19 comprised the control group. Children and adolescents with human immunodeficiency virus were enrolled. The inclusion criteria were <18 years old and consent in participating in the study. The exclusion criteria were the presence of numerous systemic comorbidities, oral lesions, the habit of smoking, alcohol consumption, and X-rays or CT scans taken within 15 days prior to sample collection. A gentle scraping was performed on the inner portion of the jugal mucosa on both sides. A total of 2,000 cells per slide were analyzed for the determination of mutagenicity parameters as follows: micronuclei, binucleation, and nuclear buds. For measuring cytotoxicity, the following metanuclear changes were evaluated: pyknosis, karyolysis, and karyorrhexis, in a double-blind manner. The repair index was also evaluated in this setting. RESULTS: The human immunodeficiency virus group showed high frequencies of micronuclei (p=0.05), binucleated cells (p=0.001), and nuclear buds (p=0.03). In the cytotoxicity parameters, represented by the cell death phases, there was an increase with statistical difference (p≤0.05) in the karyorrhexis frequency (p=0.05). Additionally, repair index was decreased in the human immunodeficiency virus group. CONCLUSION: These results indicate that human immunodeficiency virus -infected individuals undergoing antiretroviral therapy have cytogenetic changes in oral mucosal cells.

Cytogenetic changes in oral mucosa cells from individuals submitted to oral human immunodeficiency virus pre-exposure prophylaxis use

SUMMARY OBJECTIVE: The objective of this study was to evaluate cytogenetic changes in individuals submitted to oral human immunodeficiency virus pre-exposure prophylaxis use through the micronucleus test in oral mucosa. METHODS: This study consisted of 37 individuals, of whom 17 comprised the pre-exposure prophylaxis group and 20 comprised the control group. A total of 2,000 cells per slide were analyzed for the determination of micronuclei, binucleation, nuclear buds, and cytotoxicity parameters: pyknosis, karyolysis, and karyorrhexis (KR), in a double-blind manner. The repair index was also evaluated in this setting. RESULTS: In the mutagenicity parameters, the pre-exposure prophylaxis group showed increased frequencies of micronuclei (p=0.0001), binucleation (p=0.001), and nuclear buds (p=0.07). Regarding the cytotoxicity parameters, there was an increase with a statistical difference (p≤0.05) in the karyorrhexis frequency (p=0.001). Additionally, the repair system efficiency decreased in the pre-exposure prophylaxis group. CONCLUSION: These results indicate that individuals undergoing pre-exposure prophylaxis use have geno- and cytotoxicity in oral mucosal cells.

Detection of RNA-Dependent RNA Polymerase of Hubei Reo-Like Virus 7 by Next-Generation Sequencing in Aedes aegypti and Culex quinquefasciatus Mosquitoes from Brazil.

Advancements in next-generation sequencing and bioinformatics have expanded our knowledge of the diversity of viruses (pathogens and non-pathogens) harbored by mosquitoes. Hubei reo-like virus 7 (HRLV 7) was recently detected by the virome analysis of fecal samples from migratory birds in Australia. We now report the detection of RNA-dependent RNA polymerase sequences of HRLV 7 in pools of Aedes aegypti and Culex quinquefasciatus mosquitoes species from the Brazilian Amazon forest. Phylogenetic inferences indicated that all HRLV 7 strains fall within the same independent clade. In addition, HRLV 7 shared a close ancestral lineage with the Dinovernavirus genus of the Reoviridae family. Our findings indicate that HRLV 7 is present in two species of mosquitoes.

Principais fatores de internação do paciente com insuficiência renal aguda em unidade de terapia intensiva / Main factors for admission of the acute renal patient to the intensive care unit

Objetivo: Traçar um perfil sociodemográfico de pacientes com insuficiência renal aguda internados em unidade de terapia intensiva e identificar a prevalência dos principais fatores das internações, assim como as patologias de base associadas à insuficiência renal aguda. Métodos: Estudo transversal, observacional, analítico e descritivo, realizado por intermédio do levantamento dos prontuários de pacientes com insuficiência renal aguda, internados em Unidade de Terapia Intensiva, no período de janeiro de 2011 a maio de 2013. Resultados: Os pacientes tinham idade média de 56,5 anos e faixa etária predominante de 60 a 70 anos; 59% eram do sexo masculino, com predominância de raça branca (65%) e negra (34%), procedentes das cidades de Santos (35,5%) e de São Vicente (24%), sendo as principais causas de internação: desequilíbrio hidroeletrolítico (43%), insuficiência respiratória (27%) e pacientes pós-cirúrgicos (9,5%). A média de permanência na unidade de terapia intensiva foi de 20 dias. As patologias de base predominantes foram neoplasias (22,5%), diabetes (18,5%), cardiopatias (17%) e ausência de tratamentos anteriores (86,5%). Conclusão: A alta taxa de pacientes que não receberam tratamento anterior pode ser um indício de deficiência na Atenção Primária e Secundária na saúde. A alta incidência em homens brancos, na terceira idade, pode indicar a necessidade de campanhas educativas dirigidas à esta população. A altíssima taxa de mortalidade pode sugerir a necessidade de se reverem condutas médicas e de enfermagem, com o intuito de minimizar riscos a pacientes com insuficiência renal aguda.(AU)

Objective: To delineate a sociodemographic profile of patients with acute renal failure admitted to an intensive care unit, and to identify the prevalence of the main factors for admission, as well as the basic pathologies associated with acute renal failure. Methods: This is a cross-sectional, observational, analytical and descriptive study, performed through a survey of the acute renal failure patients' records who were admitted to an intensive care unit, from January 2011 to May 2013. Results: Patients' mean age was 56.5 years; the predominant age group was 60-70 years; 59% were male, with a predominance of Caucasians (65%) and black people (34%), coming predominantly from the cities of Santos (35.5%) and São Vicente (24%), with the main causes of hospitalization being water-electrolyte imbalance (43%), respiratory failure (27%), and post-surgical condition (9.5%). The average length of stay in the intensive care unit was 20 days. The predominant basic conditions were: neoplasms (22.5%), diabetes (18.5%), heart diseases (17%) and absence of previous treatments (86.5%). Conclusion: The high rate of patients who received no previous treatment may indicate deficiency in primary and secondary healthcare. The high incidence in elderly white men may indicate the need for educational campaigns directed at this population. The high mortality rate may suggest the need to review medical and nursing behavior in order to minimize risks to patients with acute renal failure.(AU)

Prevalence of depressive and anxiety symptoms in medical students in the city of Santos / Prevalência de sintomas depressivos e ansiosos em alunos de medicina na cidade de Santos

ABSTRACT Objective Ascertain the prevalence of depressive and anxiety symptoms in medical students, considering data in the literature that indicate higher vulnerability to emotional disorders in this population. Methods A descriptive cross-sectional study with a sample of 657 (98%) students. The instruments used were: questionnaire of socioeconomic-demographic characteristics, Beck Depression Inventory and Beck Anxiety Inventory. Results Predominance of the female gender (61%), aged between 17 and 30 years (98%), Catholic religion (64.2%) from the city of São Paulo (40.7%) and other cities in the state (35.7%); 30% presented depressive symptoms and 21% anxiety symptoms. Female students had higher scores both for depression (34.8%) and for anxiety (26.8%). As regards the course year, the highest rates were found in the 5th year (40.7%) for depression and in the 2nd year for anxiety (28.8%). Conclusion The data obtained in this study (30%) agreed with the literature regarding the prevalence of depressive symptoms in medical students, but this index was higher compared to the population in general (15.1% to 16.8%), and related to people in São Paulo city (18.5%). Concerning anxiety the rates found were slightly lower than those in specific literature but higher than those in literature for the population in general (8% to 18%) and in city São Paulo (16.8%). These indices indicate that the school of medicine may play a role as a predisposing and/or triggering factor in some students. The results suggest that more attention should be directed to 5th year students, who are beginning the internship period.

RESUMO Objetivo Verificar a prevalência de sintomas depressivos e ansiosos em acadêmicos de Medicina, considerando-se dados da literatura que apontam maior vulnerabilidade para distúrbios emocionais nessa população. Métodos Estudo transversal descritivo com amostra de 657 (98%) alunos, realizado na cidade de Santos, região do litoral paulista. Instrumentos utilizados questionário de características socioeconômico-demográficas, Inventário para Depressão de Beck e Inventário de Ansiedade de Beck. Resultados Predominância do gênero feminino (61%), faixa etária entre 17 e 30 anos (98%), religião católica (64,2%), procedente da cidade de São Paulo (40,7%) e de outras cidades do interior do estado (35,7%); 30% apresentaram sintomas depressivos e 21%, sintomas ansiosos. O gênero feminino obteve escores mais elevados tanto para depressão (34,8%) quanto para ansiedade (26,8%). Em relação ao ano cursado, índices mais elevados para depressão foram constatados no 5º ano (40,7%) e para ansiedade no 2º ano (28,8%). Conclusão Os dados obtidos neste estudo (30%) foram concordantes quanto à prevalência de sintomas depressivos encontrados na literatura referentes a estudantes de Medicina e mais elevados em relação à população em geral (15,1% a 16,8%) e na cidade de São Paulo (18,5%). Quanto à ansiedade, foram verificados índices ligeiramente inferiores aos da literatura específica e superiores aos da literatura na população em geral (8% a 18%) e na cidade de São Paulo (16,8%). Tais índices indicam que a escola médica pode atuar como fator predisponente ou desencadeante em alguns alunos. Resultados sugerem maior atenção para com os alunos do 5º ano, período referente ao início do internato.

Clonagem molecular do oncogene EZH2 de leucemiamieloide crônica e perspectivas terapêuticas / Molecular cloning of EZH2 oncogene in chronic myeloid leukemia andtherapeutic prospects

A Leucemia Mieloide Crônica (LMC) é uma neoplasia mieloproliferativa originada, em 95% dos casos, por uma anormalidadecitogenética que se caracteriza pela translocação recíproca entre os cromossomos 9 e 22 t(9; 22) (q34; q11), resultandono cromossomo Philadelphia (Ph). Considerando que a cura da LMC só é possível com um transplante de medulaóssea bem-sucedido e que existem casos de resistência ao inibidor de tirosina quinase Mesilato de Imatinibe, fármacode primeira escolha para o tratamento, é importante que se conheça detalhadamente os genes e proteínas alterados naLMC, favorecendo a efetividade de estratégias terapêuticas, a otimização do diagnóstico e a detecção de doença residualmínima. Dentre as novas abordagens terapêuticas para a LMC está a terapia gênica, que dependendo do gene alvo, podetambém ser eficiente para o tratamento de outras neoplasias. Dentro deste contexto, os objetivos do trabalho foram:realizar a clonagem molecular de um fragmento de DNA do gene EZH2, alvo promissor da terapia gênica, que provocaalteração epigenética na LMC e em diversas neoplasias; bem como contribuir para outros trabalhos de manipulação gênicainterespécies, os quais tem grande contribuição para a área da saúde. Para isso, o gene EZH2 foi isolado a partir do DNAgenômico de sangue periférico de pacientes com LMC e posteriormente clonado em sistemas procariotos. Dessa forma,foram realizados, definidos e comprovados os procedimentos de manipulação gênica interespécies e discutido o futuro dabiotecnologia no auxílio a pesquisas e a tratamentos, com foco na LMC.

Chronic myeloid leukemia (CML) is a myeloproliferative neoplasia, caused in 95 % of the cases by a cytogenetic abnormalitycharacterized by the reciprocal translocation between chromosomes 9 and 22 t(9; 22) (q34; q11), resulting in Philadelphiachromosome (Ph).Considering that CML cure is only possible with a successful bone marrow transplantation and that thereare resistance cases to the tyrosine kinase inhibitor Imatinib Mesylate, prescribed in first line drug treatment; it is importantto know in detail the genes and proteins that are possibly altered in CML, favoring effective therapeutic strategies, optimizeddiagnosis and minimal residual disease detection. Among the new therapeutic approaches to CML is gene therapy, which,depending on the target gene, can be efficient to other neoplasms treatment. In this context, the aims of this work were:to proceed the molecular cloning of a DNA fragment from EZH2 gene, potential target for gene therapy, that promotesepigenetic alteration in CML and in many neoplasms; as well as to contribute to studies related to interspecies gene transferthat have high contribution to health. For this reason, the EZH2 gene was isolated from peripheral blood genomic DNAfrom CML patients and cloned in prokaryotic systems. Therefore, in this study, we proceed, defined and proved the interspeciesgene transfer procedures and discussed the future of biotechnology in researches and treatments, especially in CML.

Homogenous HIV-1 subtype B quasispecies in Brazilian men and women recently infected via heterosexual transmission.

HIV has extraordinary genetic mutability, both among individuals and at the population level. However, studies of primary HIV-1 infection and serum-converters indicate that the viral population is homogeneous at the sequence level, which suggests clonal HIV transmission. It remains unclear whether this feature applies to the female population. Ten single genome amplification sequences were generated from ten individuals (five females) with recent heterosexually acquired HIV infection as determined by the serologic testing algorithm for recent HIV seroconversion. Intra-individual genetic diversity was equally low in both genders (<2 %), with mean and median variations of 0.8 and 0 %, respectively. All of the subjects were infected with clade B. Three subjects (two females) appeared to be infected by two related viral populations, and four subjects harbored non-R5 strains. Our results support the hypothesis of clonal selection for sexual transmission of HIV-1 in both genders. Future studies that generate a larger number of clones, preferably by next generation deep sequencing, are needed to confirm these results.

Detection of lamivudine-resistant variants and mutations related to reduced antigenicity of HBsAg in individuals from the cities of Santos and São Paulo, Brazil.

BACKGROUND: Continuous long-term treatment is recommended to reduce the hepatitis B virus (HBV) viral load. However, as a consequence, resistance mutations can emerge and be transmitted to other individuals. The polymerase (POL) gene overlaps the surface (S) gene. Thus, during treatment, mutations in the POL gene may lead to changes in hepatitis B surface antigen (HBsAg). The purpose of this study was to evaluate the frequency of lamivudine and vaccine escape mutations in HBsAg-positive blood donors from the city of Santos and in untreated HBV mono-infected patients from the city of São Paulo, Brazil. METHODS: HBV DNA was extracted from 80 serum samples, of which 61 were from volunteer blood donors and 19 were from untreated HBV patients. A fragment of the POL/S genes containing 593 base pairs was amplified using nested PCR. Thirty four were PCR-positive and sequencing was performed using an ABI Prism 3130 Genetic Analyzer. Alignments and mutation mapping were performed using BioEdit software. RESULTS: HBV DNA from 21 blood donors and 13 untreated patient samples were characterized using nucleotide sequencing PCR products from the POL/S genes. We were able to detect one sample with the resistance mutation to lamivudine rtM204V + rtL180M (2.94%), which was found in a volunteer blood donor that has never used antiviral drugs. The other samples showed only compensatory mutations, such as rtL80F (5.88%), rtL80V (2.94%), rtL82V + rtV207L (2.94%), rtT128P (5.88%), rtT128N/S (2.94%) and rtS219A (5.88%). We found modifications in the S gene in 14 of the 34 samples (41.16%). The mutations detected were as follows: sM133L + sI195T (2.94%), sI195M (2.94%), sP120T (2.94%), sY100S/F (2.94%), sY100C (17.64%), sI/T126P + sQ129P (2.94%), sM198I + sF183C (2.94%) and sS210R (5.88%). CONCLUSIONS: Our results suggest the transmission of lamivudine-resistant forms. Thus, the evaluation of HBV-infected subjects for lamivudine resistance would improve treatment regime. Moreover, the mutations in the S gene may impair HBsAg antigenicity and contribute to HBsAg failure detection and vaccine escape.

Environmental analyses of the parasitic profile found in the sandy soil from the Santos municipality beaches, SP, Brazil.

The environmental contamination by geohelminths represents a world public health problem and has been well documented by several authors. However, few papers describe the presence of such contamination in saline soils of coastal beaches. A study was performed on the beaches of the municipality of Santos in the period between May 2004 to April 2005 with the aim of determining the degree of contamination, and the correlation between contamination level and seasonal conditions and characteristics of the environment. Of the 2,520 samples analyzed, 18.2% (458) were contaminated, 32.3% (148) of which were localized in children's recreational areas (playgrounds). The parasite profile found in the analyzed samples indicated the presence of several zoonotic parasites: Ancylostoma larvae (82.5%), Toxocara sp. eggs (59.4%), Ancylostomidae-like eggs (37.1%), coccid oocysts (13.5%), Trichostrongylus sp. eggs and larvae, Ascaris lumbricoides eggs, (11.6%), Entamoeba sp. cysts (10.0%), Strongyloides sp. (4.8%), several free nematoids and some non-identified parasitic structures (3.3%). It was established that the highest frequency of parasitic structures occurred in the months between May and October 2004, and from February to March 2005. An increase in the diversity of parasitic forms was documented in the months between February to December 2004 and from January to April 2005, these periods having the highest rainfall.

Environmental analyses of the parasitic profile found in the sandy soil from the Santos municipality beaches, SP, Brazil / Análise ambiental do perfil de estruturas parasitárias encontradas no solo arenoso das praias do município de Santos, SP, Brasil

The environmental contamination by geohelminths represents a world public health problem and has been well documented by several authors. However, few papers describe the presence of such contamination in saline soils of coastal beaches. A study was performed on the beaches of the municipality of Santos in the period between May 2004 to April 2005 with the aim of determining the degree of contamination, and the correlation between contamination level and seasonal conditions and characteristics of the environment. Of the 2,520 samples analyzed, 18.2 percent (458) were contaminated, 32.3 percent (148) of which were localized in children's recreational areas (playgrounds). The parasite profile found in the analyzed samples indicated the presence of several zoonotic parasites: Ancylostoma larvae (82.5 percent), Toxocara sp. eggs (59.4 percent), Ancylostomidae-like eggs (37.1 percent), coccid oocysts (13.5 percent), Trichostrongylus sp. eggs and larvae, Ascaris lumbricoides eggs, (11.6 percent), Entamoeba sp. cysts (10.0 percent), Strongyloides sp. (4.8 percent), several free nematoids and some non-identified parasitic structures (3.3 percent). It was established that the highest frequency of parasitic structures occurred in the months between May and October 2004, and from February to March 2005. An increase in the diversity of parasitic forms was documented in the months between February to December 2004 and from January to April 2005, these periods having the highest rainfall.

A contaminação ambiental por geohelmintos representa um problema mundial de saúde pública e tem sido bem documentada por diversos autores. No entanto, poucos trabalhos descrevem a presença de contaminação em solos salinos de praias litorâneas. Este estudo foi realizado nas praias do município de Santos no período entre maio de 2004 a abril de 2005 com o objetivo de determinar o grau de contaminação, e possíveis correlações entre sazonalidade e características ambientais. Das 2.520 amostras analisadas, 18,2 por cento (458) estavam contaminadas, 32,3 por cento (148) das quais foram localizadas em areas de lazer das crianças(playgrounds). O perfil parasitário encontrado nas amostras analisadas indicaram a presença de vários parasitos com potencial zoonótico: larvas de Ancylostoma (82,5 por cento), ovos de Toxocara sp (59,4 por cento), ovos semelhantes aos de Ancilostomídeos (37,1 por cento), oocistos de coccídeos (13,5 por cento), ovos de Trichostrongylus sp., ovos de Ascaris lumbricoides, (11,6 por cento), cistos de Entamoeba sp. (10,0 por cento), Strongyloides sp. (4,8 por cento), vários nematóides de vida livre e algumas estruturas parasitárias não identificadas que podem corresponder a um parasita (3,3 por cento). Foi estabelecido que a maior frequência de estruturas parasitárias ocorreu nos meses entre maio e outubro de 2004, e de fevereiro a março de 2005. Um aumento na diversidade de formas parasitárias foi documentado nos meses entre fevereiro a dezembro de 2004 e de janeiro a abril de 2005, esses períodos com a maior pluviosidade.

The association between primary antiretroviral resistance and HAART virologic failure in a developing set.

Santos is a Brazilian port city with high HIV incidence, high primary antiretroviral resistance levels, high HIV-1 BF recombinants prevalence, and high rates of antiretroviral virologic failure. We evaluated factors related to virologic failure after 48 weeks of HAART in this population. We compared demographic and HIV profiles among 43 individuals with virologic failure (group 1) and 37 with virologic success (group 2) after 48 weeks of HAART initiation. The overall primary antiretroviral resistance prevalence was 31.2%; 46.5% in group 1 and 13.5% in group 2 (p < 0.005). Nine patients from group 1 and seven from group 2 were infected by F or BF strains. Fifteen individuals presented with NRTI mutations, 13 with NNRTI mutations, three with PI mutations, and five with NRTI and NNRTI mutations. No significant differences were observed in baseline viral load, CD4, clade assignment, antiretrovirals used, or demographics among groups or patients harboring resistant versus wild-type viruses. In this region, there was a high prevalence of antiretroviral resistance among long standing infected patients whose disease had progressed. This finding supports the concept that resistance testing prior to ART initiation is cost effective. The association between primary antiretroviral resistance and virologic failure may suggest that primary resistance greatly impairs antiretroviral activity.

Prevalence of potentially pathogenic free-living amoebae from Acanthamoeba and Naegleria genera in non-hospital, public, internal environments from the city of Santos, Brazil

Acanthamoeba and Naegleria species are free-living amoebae (FLA) found in a large variety of natural habitats. The prevalence of such amoebae was determined from dust samples taken from public non-hospital internal environments with good standards of cleanliness from two campuses of the same University in the city of Santos (SP), Brazil, and where young and apparently healthy people circulate. The frequency of free-living amoebae in both campuseswas 39 percent and 17 percent respectively, with predominance of the genus Acanthamoeba. On the campus with a much larger number of circulating individuals, the observed frequency of free-living amoebae was 2.29 times larger (P< 0.00005). Two trophozoite forms of Naegleria fowleri, are the only species of this genus known to cause primary amoebian meningoencephalitis, a rare and non-opportunistic infection. We assume that the high frequency of these organisms in different internal locations represents some kind of public health risk.

HIV type 1 diversity from newly diagnosed patients in Santos metropolitan area/Brazil.

HIV-1 from infected subjects has been characterized in order to provide a more accurate view of the strains that are currently found in a given region. In this report, we focused on characterizing the pol gene diversity obtained from newly diagnosed patients in Santos metropolitan area, Brazil. This region is composed of nine cities and an international port. Analysis of the 33 samples revealed that 22 strains belonged to subtype B, 4 to subtype F, and 2 to subtype C; 5 strains were B/F recombinants. Our results demonstrated that 18.2% of samples were primary antiretroviral resistance genotypic mutations, with high-level resistance to reverse transcriptase inhibitors in both subtypes B and F and in recombinant forms B/F. Our data revealed that the primary antiretroviral resistance genotypic mutations should be carefully investigated in developing countries with widespread access to antiretrovirals, such as Brazil.

Prevalence of potentially pathogenic free-living amoebae from Acanthamoeba and Naegleria genera in non-hospital, public, internal environments from the city of Santos, Brazil.

Acanthamoeba and Naegleria species are free-living amoebae (FLA) found in a large variety of natural habitats. The prevalence of such amoebae was determined from dust samples taken from public non-hospital internal environments with good standards of cleanliness from two campuses of the same University in the city of Santos (SP), Brazil, and where young and apparently healthy people circulate. The frequency of free-living amoebae in both campuses was 39% and 17% respectively, with predominance of the genus Acanthamoeba. On the campus with a much larger number of circulating individuals, the observed frequency of free-living amoebae was 2.29 times larger (P< 0.00005). Two trophozoite forms of Naegleria fowleri, are the only species of this genus known to cause primary amoebian meningoencephalitis, a rare and non-opportunistic infection. We assume that the high frequency of these organisms in different internal locations represents some kind of public health risk.

HIV type 1 pol gene diversity and antiretroviral drug resistance mutations in Santos, Brazil.

HIV-1 antiretroviral drug resistance mutations in subtype B, F, and recombinants B/F in Santos, Brazil were characterized. We studied 83 samples from individuals enrolled at the Brazilian HIV/AIDS programs from Santos. These patients have been treated with multiantiretroviral therapy. Samples were collected in 2006; RNA was extracted from plasma and used as a target to amplify the pol gene of HIV-1. PCR products were sequenced on both strands, phylogenetic analyses were performed by neighbor-joining, and recombination was evaluated by bootscan. pol gene sequencing of the samples revealed that 54 strains belonged to subtype B, 4 were subtype F, 1 was subtype C, and 24 were B/F recombinants. Recombinant break points in 20 samples are the same identified in CRF28_BF and CRF29_BF. Drug resistance mutations identified in common to subtypes B, F, and recombinants B/F were protease inhibitors M46I/L (29%), I54V (24%), A71V (22%), and V82A/F (31%); reverse transcriptase nucleoside resistance mutations M41L (52%), D67N (30%), K70R (26%), M184V (88%), L210W (29%), T215Y/I/F (65%), and K219Q/E/N (28%); and reverse transcriptase nonnucleoside resistance mutation K103N (52%). Our results suggest that, in general, the same amino acids are emerging in both subtypes B, F, and recombinant forms BF due to the selective pressure of antiretrovirals. Recombinant break points in samples are the same as identified in CRF28_BF and CRF29_BF and are recognized as important for the evolution of the local epidemic.

High levels of primary antiretroviral resistance genotypic mutations and B/F recombinants in Santos, Brazil.

This study characterized HIV-1 among antiretroviral-naïve populations presenting recent infection (RI) or long-standing infection (LSI). Sera collected from January 1999 to December 2001 at an anonymous HIV testing site in Santos, Brazil, were submitted to serologic testing algorithm for recent HIV seroconversion (STARHS). The STARHS methodology uses a combination of a sensitive and a less sensitive version of an anti-HIV enzyme immunoassay (EIA), and specimens found to be positive on the sensitive EIA and negative on the less sensitive EIA are considered to represent RI. HIV-1 V3 and pol regions of those with RI and LSI were compared. Antiretroviral resistance was defined solely by genotypic analysis. Ninety samples were evaluated representing those taken from an original cohort of 345 individuals, for whom adequate samples were available. Of 90 HIV-positive individuals, 25 presented RI. Cumulatively, 36.8% of those with RI and 25% of those with LSI presented resistance to at least one antiretroviral class. In the pol and V3 regions, 47% and 53% of those with RI presented clade B viruses and B/F recombinant viruses, respectively, whereas 56.2%, 41.7%, and 2.1% of those with LSI harbored clades B, B/F, and clade C viruses, respectively. Primary resistance and the prevalence of B/F recombinants was high in this population. Monitoring HIV-1 genetic diversity is important for developing vaccines and treatment strategies.