SARS-COV-2 detection in saliva and nasopharyngeal swabs using RT-PCR was similar

Abstract The World Health Organization has declared the widespread spread of SARS-CoV-2 and its associated disease (COVID-19) a public health emergency. The standard gold test for detecting the virus is the RT-PCR, performed from nasopharyngeal swab (NPS) samples. However, this test may be uncomfortable for the patient and requires specific training and attire from the health professional responsible for collecting the sample. Therefore, the search for alternative ways to collect samples that may be used in the diagnosis of COVID-19 is relevant. This study aimed to compare the results obtained from NPS and saliva samples. NPS and saliva samples were collected from 189 symptomatic outpatients suspected of COVID-19, who came to Piquet Carneiro Polyclinic. RNA extraction was performed using the Bio-Gene DNA/RNA Viral Extraction kit (Bioclin®). Real-time reverse transcriptase-polymerase chain reaction (RT-PCR) reactions used the Molecular SARS-CoV-2 (E / RP) kit (Bio-Manguinhos). The results indicated that 142 showed a non-detectable result (ND), while 47 showed a detectable result (D). Among the 142 "ND", 137 (94.4%) saliva samples obtained the same result, while 5 samples (3.4%) were "D". Among the 47 "D" swab samples, 35 (74.4%) showed the same result in the saliva samples. The sensitivity of the saliva test was 0.74 and the specificity was 0.97. The positive predictive value was 0.88 while the negative predictive value was 0.92. The results showed that detection of Sars-CoV-2 using saliva samples showed high sensitivity and specificity compared to nasopharyngeal swabs.

Resumo A Organização Mundial da Saúde declarou a disseminação generalizada do SARS-CoV-2 e sua doença associada (COVID-19) uma emergência de saúde pública. O teste padrão ouro para detecção do vírus é o RT-PCR, realizado a partir de amostras de swab nasofaríngeo (NPS). No entanto, esse exame pode ser desconfortável para o paciente e requer treinamento específico e vestimenta do profissional de saúde responsável pela coleta da amostra. Portanto, a busca por formas alternativas de coleta de amostras que possam ser utilizadas no diagnóstico de COVID-19 é relevante. O objetivo deste estudo foi comparar os resultados obtidos em amostras de NPS e saliva. Amostras de NPS e saliva foram coletadas de 189 pacientes ambulatoriais sintomáticos com suspeita de COVID-19, que procuraram a Policlínica Piquet Carneiro. A extração de RNA foi realizada com o kit Bio-Gene DNA / RNA Viral Extraction (Bioclin®) e as reações em tempo real da reação em cadeia da polimerase-transcriptase reversa (RT-PCR) usaram o kit Molecular SARS-CoV-2 (E / RP) (Bio-Manguinhos). Os resultados indicaram que 142 apresentaram resultado não detectável (ND), enquanto 47 apresentaram resultado detectável (D). Entre os 142 "ND", 137 (94,4%) amostras de saliva obtiveram o mesmo resultado, enquanto 5 amostras (3,4%) foram "D". Dentre as 47 amostras de swab "D", 35 (74,4%) apresentaram o mesmo resultado nas amostras de saliva. A sensibilidade do teste de saliva foi de 0,74 e a especificidade foi de 0,97. O valor preditivo positivo foi de 0,88, enquanto o valor preditivo negativo foi de 0,92. Os resultados mostraram que a detecção de Sars-CoV-2 em amostras de saliva apresentou alta sensibilidade e especificidade quando comparada com swabs nasofaríngeos.

The incidence and geographical spread of SARS-CoV-2 in Rio de Janeiro, Brazil based on RT-PCR test results

Abstract INTRODUCTION Rio de Janeiro has hardly experienced coronavirus disease. METHODS Here, 87,442 reverse transcription polymerase chain reaction (RT-PCR) test results for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) were reported among Rio de Janeiro residents (March to September 2020). RESULTS Overall, RT-PCR positivity of 44.6% decreased over time towards 20%. Positivity was greater among males (OR=1.22; 95%CI:1.19-1.26); Black (OR=1.10; 95%CI:1.02-1.19), Brown (OR=1.16; 95%CI:1.10-1.22), and indigenous people (OR=2.11; 95%CI:0.88-5.03) compared to Whites and increased with age; with epidemic spread from the capital to inland regions. CONCLUSIONS SARS-CoV-2 keeps spreading in Rio de Janeiro, and reopening of activities may fuel the epidemic.

Analysis of filaggrin 2 gene polymorphisms in patients with atopic dermatitis

Abstract Background: Polymorphisms of the filaggrin 2 gene (rs 12568784 and rs 16899374) are associated with persistent atopic dermatitis in African American patients. Filaggrin 2 is a protein with a function similar to filaggrin and also encoded in the epidermal differentiation complex on chromosome 1q21. Objective: To evaluate the polymorphisms in the filaggrin 2 gene (rs 12568784 and rs 16899374) in children and adults with atopic dermatitis and to verify the association of these with the severity of the clinical picture, presence of other allergic diseases, and socio-demographic factors. Method: The study was carried out with patients and control group. Questionnaires were used to evaluate ethnicity, sex, age, family history, scoring, atopic dermatitis (SCORAD), among other parameters. Genotyping of the filaggrin 2 gene was performed by real-time polymerase chain reaction. Results: Forty-eight patients and 83 controls were evaluated. No correlation was found between the variables studied in patients with atopic dermatitis and polymorphisms, no significant difference between the prevalence of polymorphisms in the patients and in the control group p > 0.05. Study limits: The exclusive use of self-reported ethnicity information and the sample size. Results: The results of this work can be an incentive for the study of the polymorphisms in atopic dermaititis, considering the characteristic of the Brazilian multi ethnic population. Conclusion: This is an unpublished work in Brazil and the first study in the world to have a control group to evaluate alterations in the gene of filaggrin 2.

Seroprevalence of IgG and IgM anti-SARS-CoV-2 among voluntary blood donors in Rio de Janeiro, Brazil

Background: In Brazil, mathematical models for derivingestimates and projections of COVID-19 cases have been developed without data on asymptomatic SARS-CoV-2 infection. We estimated the seroprevalence of antibodies to SARS-CoV-2 among blood donors in the State of Rio de Janeiro. Methods: Data were collected on 2,857 blood donors from April 14 to 27, 2020. We report the crude prevalence of antibodies to SARS-CoV-2, the weighted prevalence by the total state population, and adjusted prevalence estimates for test sensitivity and specificity. To establish the correlates of SARS-CoV-2 prevalence, we used logistic regression models. The analysis included period and site of blood collection, sociodemographic characteristics, and place of residence. Results: The proportion of SARS-Cov-2 positive tests without any adjustment was 4.0% (95% CI 3.3-4.7%), and the weighted prevalence was 3.8% (95% CI 3.1-4.5%). Further adjustment by test sensitivity and specificity produced lower estimates, 3.6% (95% CI 2.7-4.4%) and 3.3% (95% CI 2.6-4.1%), respectively. The variable most significantly associated with the crude prevalence was the period of blood collection: the later the period, the higher the prevalence. Regarding socio-demographic characteristics, the younger the blood donors, the higher the prevalence, and the lower the educational level, the higher the odds of a positive SARS-Cov-2 antibody. Similar results were found for the weighted prevalence. Discussion: Although our findings resulted from a convenience sample, they match some basic premises: the increasing trend over time, since the epidemic curve in the state is still on the rise; the higher prevalence among the youngest who are more likely to circulate; and the higher prevalence among the less educated as they have more difficulties in following the social distancing recommendations. Despite the study limitations, it is possible to infer that protective levels of natural herd immunity to SARS-CoV-2 are far from being reached in Rio de Janeiro. (AU)

Seroprevalence of anti-SARS-CoV-2 among blood donors in Rio de Janeiro, Brazil

ABSTRACT OBJECTIVE To estimate the seroprevalence of antibodies to SARS-CoV-2 among blood donors in the state of Rio de Janeiro, Brazil. METHODS Data were collected on 2,857 blood donors from April 14 to 27, 2020. This study reports crude prevalence of antibodies to SARS-CoV-2, population weighted prevalence for the state, and prevalence adjusted for test sensitivity and specificity. Logistic regression models were used to establish the correlates of SARS-CoV-2 prevalence. For the analysis, we considered collection period and site, sociodemographic characteristics, and place of residence. RESULTS The proportion of positive tests for SARS-Cov-2, without any adjustment, was 4.0% (95%CI 3.3-4.7%), and the weighted prevalence was 3.8% (95%CI 3.1-4.5%). We found lower estimates after adjusting for test sensitivity and specificity: 3.6% (95%CI 2.7-4.4%) for the non-weighted prevalence, and 3.3% (95%CI 2.6-4.1%) for the weighted prevalence. Collection period was the variable most significantly associated with crude prevalence: the later the period, the higher the prevalence. Regarding sociodemographic characteristics, the younger the blood donor, the higher the prevalence, and the lower the education level, the higher the odds of testing positive for SARS-Cov-2 antibody. We found similar results for weighted prevalence. CONCLUSIONS Our findings comply with some basic premises: the increasing trend over time, as the epidemic curve in the state is still on the rise; and the higher prevalence among both the youngest, for moving around more than older age groups, and the less educated, for encountering more difficulties in following social distancing recommendations. Despite the study limitations, we may infer that Rio de Janeiro is far from reaching the required levels of herd immunity against SARS-CoV-2.

Genetic ancestry of patients with porphyria cutanea tarda in a country with mixed races: a cross-sectional study (Rio de Janeiro - Brazil)

Abstract: Porphyria cutanea tarda has a complex etiology with genetic factors not completely elucidated. The miscegenation of the Brazilian population has important implications in the predisposition to diseases. There are no studies concerning the genetic ancestry of patients with porphyria cutanea tarda from a mixed population. Thirty patients living in Rio de Janeiro with sporadic porphyria cutanea tarda were studied for the genetic ancestry through informative markers - INDELS. There was a significant predominance of European ancestry across the sample of patients with porphyria cutanea tarda (70.2%), and a small contribution of African and Amerindian ancestry, 20.1% and 10.9%, respectively.

DRESS: relato de caso com estudo genético / DRESS syndrome: case report with genetic testin

A síndrome de hipersensibilidade a drogas com eosinofilia e sintomas sistêmicos (DRESS) é uma rara reação adversa a drogas com potencial de morte e sequelas em longo prazo. Os anticonvulsivantes aromáticos estão entre os medicamentos mais relacionados. Relatamos um caso de DRESS em associação com o alelo HLA-A*31:01, destacando aspectos clínico-laboratoriais, abordagem diagnóstica e acompanhamento ambulatorial de sequelas tardias. Homem com 69 anos, natural do Japão, internado com suspeita clínica de DRESS. Havia iniciado carbamazepina 4 semanas antes do rash cutâneo para tratamento de epilepsia. Apresentou biópsia cutânea compatível com farmacodermia. O paciente foi tratado com prednisolona por 4 meses. A tipagem HLA-A-B-DRB1 por PCR-RSSO (ONE LAMBDA) e SSP alelo específico revelou HLA relacionado a reações de hipersensibilidade à carbamazepina. O teste de contato realizado com carbamazepina a 10% no primeiro ano após a reação foi positivo. A restrição futura da classe de anticonvulsivantes aromáticos foi recomendada. Oito meses após a aparente resolução clínica da DRESS, o paciente desenvolveu aumento dos anticorpos antitireoideanos e doença de Hashimoto. Treze meses após a o início da reação, foi observado aumento nos títulos de FAN, sem manifestações clínicas. Este relato de caso descreve aspectos clínico-laboratoriais típicos de DRESS relativos ao diagnóstico clínico-laboratorial e histopatológico, bem como evolução clínica em curto e longo prazos. A abordagem farmacogenética e o teste de contato foram importantes para a confirmação da imputabilidade da carbamazepina na etiologia da DRESS.

Drug reaction with eosinophilia and systemic symptoms (DRESS syndrome) is a rare, potentially fatal adverse reaction to drugs that may have long-term sequelae. Aromatic anticonvulsants are among the drugs most commonly associated with DRESS. We report a case of DRESS associated with allele HLA-A*31:01, with emphasis on clinical and laboratory findings, the diagnostic approach adopted, and outpatient follow-up of late sequelae. A 69-year old Japanese male patient was admitted with a clinical suspicion of DRESS. He had started carbamazepine treatment for epilepsy 4 weeks before the rash. He presented skin biopsy compatible with pharmacodermia. The patient was treated with prednisolone for 4 months. HLA-A-B-DRB1 typing using the PCRRSSO technique (ONE LAMBDA) and specific SSP allele revealed HLA related to hypersensitivity reactions to carbamazepine. The skin test performed with carbamazepine 10% on the first day after the reaction resulted positive. Future restriction of aromatic anticonvulsants was recommended. Eight months after the apparent clinical resolution of DRESS, the patient showed increased levels of antithyroid antibodies and Hashimoto disease. Thirteen months after the onset of the reaction, increased FAN results were observed, with no clinical manifestations. This case report describes clinical and laboratory aspects of DRESS related to clinical, laboratory, and histopathological diagnosis, as well as clinical evolution in the short and long terms. The pharmacogenetic approach and the skin test were important to confirm the imputability of carbamazepine in the etiology of DRESS.

Exposure source prevalence is associated with gender in hepatitis C virus patients from Rio de Janeiro, Brazil

BACKGROUND Hepatitis C virus (HCV) infection is a worldwide public health problem. A characterisation of the differences in exposure sources among genders will enable improvements in surveillance actions. METHODS Exposure data were obtained for 1180 confirmed HCV cases Brazil's mandatory reporting to epidemiological surveillance, which was directed by a reference laboratory in Rio de Janeiro, Brazil. The Chi-square test (χ2) was used to assess the associations between exposure sources and gender. The prevalence ratio (PR) was calculated for exposures that showed an association. RESULTS The results showed 57.7% cases were female, and associations with snorting drugs, sexual activity, surgery, aesthetic procedures, blood transfusions, and educational level were observed (p < 0.001). Men showed 2.53 (1.33-3.57), 4.83 (3.54-6.59), and 2.18 (1.33-3.57) times more exposure to sniffing drugs, risky sex and higher levels of education, respectively, than women. Women demonstrated 4.46 (3.21-6.21), 1.94 (1.43-2.63), and 3.10 (2.09-4.61) times more exposure to surgery, aesthetic procedures, and blood transfusions, respectively, than men. CONCLUSION Our results showed differences in risk behaviours associated with gender among HCV carriers. These data are likely to significantly influence clinical practice regarding the adoption of specific approaches for counselling and control policies to prevent the emergence of new cases and break the chain of transmission of the virus.

HBV and HCV serological markers in health professionals and users of the Brazilian Unified Health System network in the city of Resende, Rio de Janeiro, Brazil / Marcadores sorológicos para HBV e HBC em profissionais de saúde e usuários da rede do Sistema Único de Saúde no Município de Resende, Rio de Janeiro, Brasil

ABSTRACT Introduction: Infections caused by the hepatitis B virus (HBV) and hepatitis C virus (HCV) are a major public health problem. Objectives: The study aimed to detect HBsAg, anti-HBc, anti-HBs and anti-HCV among health professionals and users of the Brazilian Unified Health System [Sistema Único de Saúde (SUS)] in the city of Resende, Rio de Janeiro, and to describe the sociodemographic profile and background of exposure. Methods: A total of 585 samples were collected between May and June 2014, obtained from the Brazilian Notifiable Diseases Surveillance System [Sistema de Informação de Agravos de Notificação (SINAN)] data, which were tested for HBsAg, anti-HBc, anti-HBs and anti-HCV. Results: The predominant age group observed was 30-44 years (n = 277; 47.3%), 54.87% (n = 321) were female and 271 (46.32%) self declared skin colour/ethnicity white. The married participants were 262 (44.78%), 42.22% graduated from high school (n = 247) and 174 were health professionals (29.74%). Four participants were anti-HCV reagents and 18 were anti-HBc reagents. From these, 15 participants were reactive for anti-HBs antibodies. Among health professionals, 68.8% were anti-HBs positive. And 63.9% of participants declared to be vaccinated against hepatitis B. Conclusion: The prevalence of 0.68% for HCV and 3.08% for anti-HBc are below that detected in the Southeast region from the last census in the capitals of Brazil. There is still a reduced acceptance among health professionals for HBV and HCV screening.

RESUMO Introdução: As infecções causadas pelo vírus da hepatite B (VHB) e C (VHC) constituem grave problema de saúde pública. Objetivos: O estudo visou detectar os marcadores HBsAg, anti-HBc, anti-HBs e anti-VHC em profissionais de saúde e usuários do Sistema Único de Saúde (SUS) no município de Resende, Rio de Janeiro, bem como descrever o perfil sociodemográfico e os antecedentes de exposição. Métodos: Foram avaliadas 585 amostras entre maio e junho de 2014, obtidas dos dados do Sistema de Informação de Agravos de Notificação (SINAN). Elas foram testadas para HBsAg, anti-HBc, anti-HBs e anti-VHC. Resultados: A faixa etária predominante observada foi de 30-44 anos (n = 277; 47,3%); 54,87% (n = 321) eram do sexo feminino e 271 (46,32%) se autodeclararam de cor da pele/etnia branca. Os participantes casados foram 262 (44,78%); 42,22% tinham o ensino médio (n = 247) e 174 eram profissionais de saúde (29,74%). Quatro participantes eram anti-VHC reagentes e 18, reagentes para anti-HBc. Destes, 15 eram anti-HBs reagentes (aHBs+). Nos profissionais de saúde, 68,8% possuem aHbs+. Em relação à vacinação contra hepatite B, 63,9% declararam possuí-la. Conclusão: As prevalências 0,68% de VHC e de 3,08% de anti-HBc estão abaixo da detectada na região Sudeste no último censo nas capitais do Brasil. Há ainda reduzida adesão dos profissionais de saúde à testagem para VHB e VHC.

Analysis of mutations in the PIK3CA and FGFR3 genes in verrucous epidermal nevus / Analise de mutacoes nos genes PIK3CA e FGFR3 em caso de nevo epidermico verrucoso

Verrucous epidermal nevi are congenital hamartomas composed of keratinocytes and may occur alone or in association with developmental abnormalities. A close relationship between variations in the PIK3CA and FGFR3 genes and the appearance of nevi has been recently reported. Based on that, we performed molecular assays for the identification of E542K, E545G/K and H1047R mutations in the PIK3CA gene and of the R248C mutation in the FGFR3 gene. Interestingly, during the amplification process, we did not observe the PCR product of exon 9 of the PIK3CA gene, a region comprising amino acids 542-545. This strongly suggests the occurrence of a microdeletion of that region and indicates a possible allelic variant, which has not yet being described in the literature.

Os nevos epidérmicos verrucosos são hamartomas congênitos compostos por queratinócitos, que podem ocorrer isolados ou associados a alterações do desenvolvimento. Com a recente observação de uma relação estreita entre variações nos genes PIK3CA e FGFR3 e o aparecimento do nevo, realizamos ensaios moleculares para identificação das mutações E542K, E545G/K e H1047R do gene PIK3CA, e R248C do gene FGFR3. Interessantemente, durante o processo de amplificação não observamos o produto da PCR do exon 9 do gene PIK3CA, região que compreende aos aminoácidos 542-545, sugerindo fortemente a ocorrência de uma microdeleção da região e indicando uma provável variante alélica ainda não descrita na literatura.

A resposta oxidativa em corações de camundongos é modulada por background genético / The oxidative response of mouse hearts is modulated by genetic background

FUNDAMENTO: o tabagismo apresenta importante papel sobre as doenças cardiovasculares, entretanto permanecem pouco compreendidos os motivos pelos quais alguns seres humanos as desenvolvem e outros não. OBJETIVO: nosso objetivo foi analisar o perfil redox do coração de diferentes linhagens de camundongos após exposição à fumaça de cigarro. MÉTODOS: camundongos machos suíços (n = 10), C3H (n = 10), BALB/c (n = 10) e C57BL/6 (n = 10) foram expostos à fumaça de cigarro (12 cigarros/dia), enquanto os respectivos controles (n = 10) ao ar ambiente por 60 dias. Após sacrifício, o coração foi retirado para análises bioquímicas. RESULTADOS: embora o conteúdo de malondialdeído não tenha aumentado em nenhum grupo, a atividade da catalase diminuiu no grupo suíço (p < 0,05), BALB/c (p < 0,05) quando comparados aos respectivos grupos-controle, enquanto a mieloperoxidase diminuiu no grupo C3H (p < 0,05) e C57BL/6 (p < 0,001) quando comparados aos respectivos grupos controle. O conteúdo de glutationa reduzida diminuiu nos grupos suíço, C3H, C57BL/6 (p < 0,05) e no grupo BALB/c (p < 0,001) quando comparados com os respectivos controles. Observamos aumento do conteúdo da glutationa oxidada no grupo Suíço (p < 0,05) e diminuição nos grupos C3H (p < 0,05) e BALB/c (p < 0,001) quando comparados aos respectivos grupos-controle. A razão glutationa reduzida/ glutationa oxidada apresentou redução nos grupos suíço e C57BL/6 (p < 0.05) quando comparados aos grupos controle. CONCLUSÃO: o background genético nos camundongos pode influenciar na resposta antioxidante após a exposição à fumaça de cigarro e parece ser um fator determinante para o desequilíbrio redox no suíço e C57BL/6. Compreender as respostas antioxidantes e do background genético C3H e BALB/c podem fornecer importantes informações quanto à resistência cardíaca a fumaça de cigarro.

BACKGROUND: Smoking plays an important role in cardiovascular diseases. However, the reasons why some individuals develop those diseases and others do not remain to be explained. OBJECTIVE: This study aimed at assessing the redox profile of the heart of different mouse strains after exposure to cigarette smoke. METHODS: Male mice of the Swiss (n = 10), C3H (n = 10), BALB/c (n = 10) and C57BL/6 (n = 10) strains were exposed to cigarette smoke (12 cigarettes/day), while their respective controls (n = 10) were exposed to ambient air for 60 days. After being euthanized, their heart was removed for biochemical analyses. RESULTS: Although the malondialdehyde content did not increase in any of the groups, catalase activity decreased in the Swiss (p < 0.05) and BALB/c (p < 0.05) strain mice as compared with their respective control groups, while myeloperoxidase decreased in the C3H (p < 0.05) and C57BL/6 (p < 0.001) strain mice as compared with their respective control groups. The reduced glutathione content decreased in the Swiss, C3H, C57BL/6 (p < 0.05) and BALB/c (p < 0,001) strain mice as compared with their respective control groups. Regarding reduced glutathione content, an increase was observed in the Swiss strain mice (p < 0.05), while a decrease was observed in the C3H (p < 0.05) and BALB/c (p < 0.001) strain mice as compared with their respective control groups. The reduced glutathione/reduced glutathione ratio showed a reduction in the Swiss and C57BL/6 (p < 0.05) strain mice as compared with their respective control groups. CONCLUSIONS: The genetic background of mice can influence the antioxidant response after exposure to cigarette smoke and seems to be a determinant factor for redox imbalance in Swiss and C57BL/6 strain mice. Understanding antioxidant responses and genetic background of C3H and BALB/c strain mice might provide important information regarding cardiac resistance to cigarette smoke.

Ventilação mecânica com baixo volume corrente e estresse oxidativo em pulmões saudáveis de camundongos / Low tidal volume mechanical ventilation and oxidative stress in healthy mouse lungs

OBJETIVO: A ventilação mecânica (VM) por si própria pode contribuir diretamente para a lesão pulmonar. Assim, o objetivo do presente estudo foi investigar biomarcadores precoces relacionados ao equilíbrio oxidantes/antioxidantes, estresse oxidativo e inflamação causados por VM de curta duração em pulmões de camundongos saudáveis. MÉTODOS: Vinte camundongos C57BL/6 machos foram randomicamente divididos em dois grupos: VM, submetidos a VM com baixo volume corrente (V T, 6 mL/kg) por 30 min; e respiração espontânea (RE), utilizados como controles. Amostras de homogeneizados de pulmão foram testados quanto à atividade de enzimas antioxidantes, peroxidação lipídica e expressão de TNF-α. RESULTADOS: Comparados ao grupo RE, houve uma redução significativa na atividade de superóxido dismutase (≈35 por cento; p < 0,05) e aumento da atividade de catalase (40 por cento; p < 0,01), glutationa peroxidase (500 por cento; p < 0,001) e mieloperoxidase (260 por cento; p < 0,001), ao passo que a razão glutationa reduzida/glutationa oxidada foi menor (≈50 por cento; p < 0,05), e houve um aumento na atividade de expressão de TNF-α no grupo VM. O dano oxidativo, analisado como peroxidação lipídica, também aumentou no grupo VM (45 por cento; p < 0.05). CONCLUSÕES: Nossos resultados demonstraram que VM de curta duração com baixa V T pode contribuir diretamente para a lesão pulmonar, gerando estresse oxidativo e inflamação em pulmões de camundongos saudáveis.

OBJECTIVE: Mechanical ventilation (MV) itself can directly contribute to lung injury. Therefore, the aim of the present study was to investigate early biomarkers concerning oxidant/antioxidant balance, oxidative stress, and inflammation caused by short-term MV in healthy mouse lungs. METHODS: Twenty male C57BL/6 mice were randomly divided into two groups: MV, submitted to low tidal volume (V T, 6 mL/kg) MV for 30 min; and spontaneous respiration (SR), used as controls. Lung homogenate samples were tested regarding the activity of various antioxidant enzymes, lipid peroxidation, and TNF-α expression. RESULTS: In comparison with the SR group, the MV group showed a significant decrease in the activity of superoxide dismutase (≈35 percent; p < 0.05), together with an increase in the activity of catalase (40 percent; p < 0.01), glutathione peroxidase (500 percent; p < 0.001), and myeloperoxidase (260 percent; p < 0.001), as well as a reduction in the glutathione/oxidized glutathione ratio (≈50 percent; p < 0.05) and an increase in TNF-α expression in the MV group. Oxidative damage, assessed by lipid peroxidation, was also greater in the MV group (45 percent; p < 0.05). CONCLUSIONS: Our results show that short-term low V T MV can directly contribute to lung injury, generating oxidative stress and inflammation in healthy mouse lungs.

Association of hepatitis C virus NS5B variants with resistance to new antiviral drugs among untreated patients

Mutations located in the 109-amino acid fragment of NS5B are typically associated with resistance to interferon (IFN) and ribavirin (RIB) and to new antiviral drugs. The prevalence of these mutations was examined in 69 drug-naïve individuals with hepatitis C virus (HCV) infections in Rio de Janeiro, Brazil. Mutations related to non-response to IFN/RIB were observed in all subtypes studied (1a, 1b, 2b, 3a and 4). The most common mutation was Q309R, present in all subtypes, except subtype 2b with frequency above 20 percent. D244N was detected only in subtype 3a and A333E was detected only in subtype 2b. We did not detect the S282T, S326G or T329I mutations in any of the samples analysed. Of note, the C316N mutation, previously related to a new non-nucleoside compound (HCV796 and AG-021541), was observed in only eight of 33 (24 percent) samples from subtype 1b. Site 316 was under positive selection in this HCV variant. Our data highlight the presence of previously described resistance mutations in HCV genotypes from drug-naïve patients.

Estudo da frequência dos alelos de HLA-DRB1 em pacientes brasileiros com artrite reumatoide / Study of the frequency of HLA-DRB1 alleles in Brazilian patients with rheumatoid arthritis

Os alelos HLA-DRB1, que codificam uma sequência de aminoácidos (QKRAA/QRRAA/RRRAA) nas posições 70 a 74 da terceira região hipervariável da cadeia β1 do gene DRB1, denominada epítopo compartilhado (EC), estão associados a maior suscetibilidade e gravidade para artrite reumatoide (AR) em diversas populações. OBJETIVO: Determinar a frequência dos alelos HLA-DRB1 em pacientes brasileiros com AR, e sua associação a fator reumatoide (FR) e anticorpos antipeptídeos citrulinados (ACPA). MATERIAL E MÉTODOS: Foram incluídos 412 pacientes com AR e 215 controles. A tipificação HLA-DRB1 foi realizada pela reação em cadeia da polimerase (PCR) usando primers específicos e hibridização com oligonucleotídeos de sequência específica (SSOP). A pesquisa de ACPA foi determinada pela técnica de ELISA, e a do FR por nefelometria. Para análises estatísticas foram utilizados os testes do qui-quadrado e t de Student e a regressão logística. RESULTADOS: Alelos HLA-DRB1*04:01, *04:04 e *04:05 associaram-se à AR (P < 0,05)); a despeito do amplo intervalo de confiança, vale a pena ressaltar a associação observada entre o alelo DRB1*09:01 e a doença (P < 0,05). Alelos HLA-DRB1 EC+ foram observados em 62,8 por cento dos pacientes e em 31,1 por cento do grupo-controle (OR 3,62; P < 0,001) e estiveram associados a ACPA (OR 2,03; P < 0,001). Alelos DRB1-DERAA mostraram efeito protetor para AR (OR 0,42; P < 0,001). CONCLUSÃO: Em uma amostra de pacientes brasileiros com AR de etnia majoritariamente mestiça, alelos HLA-DRB1 EC+ estiveram associados à suscetibilidade à doença e à presença de ACPA.

The HLA-DRB1 alleles encoding an amino acid sequence (QKRAA/QRRAA/RRRAA) at position 70 74 of the third hypervariable region of the β1 chain of the HLA-DRB1 gene, called shared epitope (SE), are associated with increased susceptibility to and severity of rheumatoid arthritis (RA) in different populations. OBJECTIVE: To determine the frequency of HLA-DRB1 alleles in Brazilian patients with RA and their association with rheumatoid factor (RF) and anti-citrullinated peptide antibodies (ACPA). METHODS: Four hundred and twelve patients with RA (ACR 1987) and 215 controls were included. HLA-DRB1 typing was performed by use of polymerase chain reaction (PCR) with specific primers and hybridization with sequence-specific oligonucleotide probe (SSOP). ACPA was measured by use of the ELISA technique and RF by nephelometry. The statistical analysis comprised the chi-square and Student t tests and logistic regression. RESULTS: HLA-DRB1*04:01, *04:04, *04:05 alleles were associated with RA (P < 0.05); despite the wide confidence interval, it is worth noting the association between the DRB1*09:01 allele and RA (P < 0.05). HLA-DRB1 SE+ alleles were observed in 62.8 percent of the patients and in 31.1 percent of controls (OR 3.62; P < 0.001) and were associated with ACPA (OR 2.03; P < 0.001). DRB1-DERAA alleles showed a protective effect against RA (OR 0.42; P < 0.001). CONCLUSION: In a sample of Brazilian patients with RA, most of whom of mixed heritage, HLA-DRB1 SE+ alleles were associated with susceptibility to disease and presence of ACPA.

Early proliferation of bone marrow mononuclear cells on collagen membrane, bone graft and tooth cementum / Proliferación temprana de células mononucleares de médula óseasobre membrana de colágeno, injerto óseo y cemento dentario

The ultimate goal of periodontal therapy is to repair the damaged periodontal supporting tissues, permitting regeneration of the periodontal ligament. However, the cell response, the supportive matrix and the bioactive molecules use have not yet been well established. Bone marrow mononuclear cells were extracted from rat femurs and tibiae and cultured on a cross-linked collagen membrane, bone graft, or molar tooth to compare cell attachment and early proliferation on these materials. Cell attachment was quantified by light microscopy at 24, 48 and 72h, and cell proliferation was observed under a SEM after 72h. After 24h, the number of cells on bone graft was similar to that of the control and more than twice compared to collagen membrane (q=7.473 p<0.001) and 1.75 times greater than with tooth cementum (q=5.613 p<0.01). However, the number of cells close to bone graft decreased in the second day compared to the control. SEM examination revealed a significant decrease in the number of cells that attached and proliferated on tooth and bone graft when compared with membrane. The results showed that bone marrow mesenchymal cells offer great potential for colonize a collagen membrane.

El objetivo último de la terapia periodontal es reparar el daño tejidos periodontales de soporte, permitiendo la regeneración del ligamento periodontal. Sin embargo, la respuesta de la célula, la matriz de apoyo y las moléculas bioactivas aún no han sido bien establecidas. Células mononucleares de la médula ósea se extrajeron del fémur y fibula de rata, y fueron cultivadas sobre un reticulado de membrana de colágeno, de injerto de hueso o de un diente molar para comparar la adhesión celular y la proliferación temprana sobre estos materiales. La adhesión celular fue cuantificada por microscopía de luz a las 24, 48 y 72h, y la proliferación celular fue observada bajo MEB después de 72h. Después de 24 horas, el número de células sobre el injerto de hueso fue similar a la del control y más del doble en comparación con la membrana de colágeno (q=7,473 p<0,001) y 1,75 veces mayor que con el cemento dental (q=5,613 p<0,01). Sin embargo, el número de células cerca del injerto óseo disminuyó el segundo día en comparación con el control. El examen al MEB reveló una disminución significativa en el número de células que se unen y proliferan sobre los dientes y el injerto óseo en comparación con la membrana. Los resultados mostraron que las células mesenquimales de la médula ósea tienen un gran potencial para colonizar la membrana de colágeno.

Effects of oral nicotine on rat liver stereology

Nicotine is the more abundant component in cigarette smoke. Because nicotine is first metabolized in the liver, our aim was to investigate the effects of nicotine on this organ by biochemical and stereological methods. Male Wistar rats were treated with oral nicotine (ON) diluted in drinking water during 32 days. The control group was treated with drinking water in the same period. Rats were sacrificed 24 hours after last day, the blood was collected and the liver was removed. Lipidogram was performed by enzymatic method and collagen fibers, fat globules and hepatocytes were count in the liver by stereological methods. We observed in control group preserved hepatocytes, with no presence of inflammatory cells. However in the ON group was possible to observe varied size of hepatocytes with cloudy cellular limits and histoarchitecture loss. Capillaries were fully of red blood cells. We observed also an increase of fat globules with small size. We observed in leucogram a reduction of leukocytes number (lymphocytes, neutrophils and monocytes) in the ON group in comparison to control group. The lipidogram showed an increase of triglycerides and total cholesterol for ON group when compared to control group. Moreover, a reduction of HDL- cholesterol was observed in ON group when compared to control group. Numerical density of hepatocytes was lesser in ON group when compared to control group. We suggest harmful effects of oral nicotine in liver induced by toxic mechanism with reduction of hepatocytes number and disturbance in lipid metabolism.

La nicotina es el componente más abundante en el humo del cigarrillo. Primero porque la nicotina es metabolizada en el hígado, nuestro objetivo fue investigar los efectos de la nicotina sobre este órgano por métodos bioquímicos y estereológicos. Hombre rata Wistar fueron tratados con la nicotina oral (NO) diluida en el agua potable durante 32 días. El grupo control fue tratado con agua potable en el mismo período. Las ratas se sacrificaron 24 horas después del último día, se recogió la sangre y el hígado fue retirado. Lipidogram se realizó por el método enzimático y fibras de colágeno, grasa y hepatocitos se cuentan en el hígado mediante métodos estereológicos. Hemos observado en el grupo control hepatocitos conservados, sin presencia de células inflamatorias. Sin embargo en el grupo ON fue posible observar variado tamaño de hepatocitos con nubes y claros límites celulares y histoarchitecture pérdida. Capilares están plenamente de los glóbulos rojos. Se observó también un aumento de grasa con pequeño tamaño. Hemos observado en leucogram una reducción del número de glóbulos blancos (linfocitos, neutrófilos y monocitos) en el grupo ON, en comparación con el grupo control. El lipidogram mostró un aumento de los triglicéridos y de colesterol total ON grupo comparado con el grupo control. Por otra parte, una reducción del HDL-colesterol se observó en el grupo ON, en comparación con el grupo control. Densidad numérica de los hepatocitos fue menor en el grupo ON, en comparación con el grupo control. Sugerimos oral efectos nocivos de la nicotina en el hígado inducido por tóxicos con mecanismo de reducción de número de hepatocitos y alteraciones en el metabolismo lipídico.

Estudo imunohistoquímico do remodelamento pulmonar em camundongos expostos à fumaça de cigarro / Immunohistochemical study of lung remodeling in mice exposed to cigarette smoke

OBJETIVO: Muitos estudos sobre enfisema são realizados com exposição de animais à fumaça de cigarro durante um longo tempo, focando o tipo de célula envolvida no desequilíbrio protease/antiprotease e a degradação da matriz extracelular. A expressão aumentada de metaloproteinases no enfisema está associado com citocinas e evidências sugerem um papel importante da metaloproteinase de matriz-12 (MMP-12). Nosso objetivo foi estudar a detecção de inibidor tissular de metaloproteinase-2 (TIMP-2), fator de necrose tumoral alfa (TNF-α) e interleucina-6 (IL-6) por métodos imunohistoquímicos no pulmão de camundongos. MÉTODOS: Camundongos C57BL/6 machos foram expostos 3 vezes ao dia a fumaça de 3 cigarros por um período de 10, 20, 30 ou 60 dias através de uma câmara de inalação (grupos CS10, CS20, CS30 e CS60, respectivamente). O grupo controle foi exposto às mesmas condições ao ar ambiente. RESULTADOS: Nós observamos um aumento progressivo de macrófagos alveolares no lavado broncoalveolar dos grupos expostos. O diâmetro alveolar médio, um indicador de destruição alveolar, aumentou em todos os grupos expostos quando comparado ao grupo controle. O índice imunohistoquímico (II) para MMP-12 aumentou nos grupos CS10, CS20 e CS30 em paralelo a uma redução do II para TIMP-2 nos grupos CS10, CS20 e CS30. O II para as citocinas TNF-α e IL-6 aumentou em todos os grupos expostos quando comparado ao grupo controle. Enfisema foi observado no grupo CS60, com alterações na densidade de volume de fibras colágenas e elásticas. CONCLUSÕES: Estes achados sugerem que a fumaça de cigarro induz enfisema com uma participação importante do TNF-α e da IL-6 sem a participação de neutrófilos.

OBJECTIVE: Various studies of emphysema involve long-term exposure of animals to cigarette smoke, focusing on the cell type involved in the protease/antiprotease imbalance and on extracellular matrix degradation. In emphysema, increased expression of metalloproteinases is associated with cytokines, and evidence suggests that the matrix metalloproteinase-12 (MMP-12) plays an important role. Our objective was to investigate tissue inhibitor of metalloproteinase-2 (TIMP-2), tumor necrosis factor-alpha (TNF-α) and interleukin-6 (IL-6) detection by immunohistochemical methods in mouse lung. METHODS: Male C57BL/6 mice were exposed 3 times a day to smoke of 3 cigarettes over a period of 10, 20, 30 or 60 days in an inhalation chamber (groups CS10, CS20, CS30 and CS60, respectively). Controls were exposed to the same conditions in room air. RESULTS: A progressive increase in the number of alveolar macrophages was observed in the bronchoalveolar lavage fluid of the exposed mice. The mean linear intercept, an indicator of alveolar destruction, was greater in all exposed groups when compared to control group. In the CS10, CS20 and CS30 mice, the immunohistochemical index (II) for MMP-12 increased in parallel with a decrease in II for TIMP-2 in the CS10, CS20 and CS30 mice. The II for the cytokines TNF-α and IL-6 was greater in all exposed groups than in the control group. Emphysema, with changes in volume density of collagen and elastic fibers, was observed in the CS60 group. CONCLUSIONS: These findings suggest that cigarette smoke induces emphysema with major participation of TNF-α and IL-6 without participation of neutrophils.

Efeitos da hiperóxia sobre o pulmão de ratos Wistar / Effects of hyperoxia on Wistar rat lungs

OBJETIVO: Avaliar a repercussão da elevada concentração de oxigênio (hiperóxia) em um curto período de tempo no pulmão de ratos Wistar. MÉTODOS: Os animais foram divididos em grupos O10', O30', O90', ou seja, ratos expostos à hiperóxia por 10', 30' e 90', respectivamente, e no grupo controle (GC), exposto ao ar ambiente. Os animais foram sacrificados 24 h após a exposição. O lavado broncoalveolar foi realizado e os pulmões foram retirados para análise histológica e estereológica. RESULTADOS: Observamos um aumento do número de macrófagos (2169,9 ± 118,0, 1560,5 ± 107,0 e 1467,6 ± 39,0) e neutrófilos (396,3 ± 35,4, 338,4 ± 17,3 e 388,7 ± 11,7), concomitante a um aumento do dano oxidativo (143,0 ± 7,8 por cento, 180,4 ± 5,6 por cento e 235,0 ± 13,7 por cento) nos grupos O10', O30' e O90', respectivamente, quando comparados ao GC (781,3 ± 78,3 por cento, 61,6 ± 4,2 por cento e 100,6 ± 1,7 por cento). Na análise histológica e estereológica foram observados alvéolos e septos normais no GC (83,51 ± 1,20 por cento e 15 ± 1,21 por cento), no grupo O10' (81,32 ± 0,51 por cento e 16,64 ± 0,70 por cento) e no grupo O30' (78,75 ± 0,54 por cento e 17,73 ± 0,26 por cento). Entretanto, no grupo O90' foi notado um influxo de células inflamatórias nos alvéolos e nos septos alveolares. Hemácias extravasaram do capilar para o alvéolo (59,06 ± 1,22 por cento), com evidências de congestão, hemorragia e edema de septo (35,15 ± 0,69 por cento). CONCLUSÃO: Os resultados indicam que a hiperóxia induziu uma ação lesiva no grupo O90' sobre o parênquima pulmonar, com repercussões de dano oxidativo e infiltrado inflamatório.

OBJECTIVE: To study the effects of short-term exposure to high oxygen concentrations (hyperoxia) on Wistar rat lungs. METHODS: Animals were divided into three groups exposed to hyperoxia for 10', 30' and 90' (O10', O30', O90', respectively), together with a control group (exposed to room air). The animals were sacrificed 24 h after exposure. Bronchoalveolar lavage was performed, and the lungs were removed for histological and stereological analysis. RESULTS: In the O10', O30', and O90' groups, respectively and in comparison with the controls, we observed an increase in the numbers of macrophages (2169.9 ± 118.0, 1560.5 ± 107.0, and 1467.6 ± 39.0 vs. 781.3 ± 78.3) and neutrophils (396.3 ± 35.4, 338.4 ± 17.3, and 388.7 ± 11.7 vs. 61.6 ± 4.2), concomitant with an increase in oxidative damage (143.0 ± 7.8 percent, 180.4 ± 5.6 percent, and 235.0 ± 13.7 vs. 100.6 ± 1.7 percent). The histological and stereological analyses revealed normal alveoli and alveolar septa in the controls (83.51 ± 1.20 percent and 15 ± 1.21 percent), in the O10' group (81.32 ± 0.51 percent and 16.64 ± 0.70 percent), and in the O30' group (78.75 ± 0.54 percent and 17.73 ± 0.26 percent). However, in the O90' group, inflammatory cell infiltration was observed in the alveoli and alveolar septa. Red blood cells extravasated from capillaries to the alveoli (59.06 ± 1.22 percent), with evidence of congestion, hemorrhage, and septal edema (35.15 ± 0.69 percent). CONCLUSION: Hyperoxia for 90' caused injury of the lung parenchyma, resulting in oxidative damage and inflammatory cell infiltration.

Associação de suscetibilidade dos alelos HLA-DRB1 e HLADQB1 com a periodontite agressiva localizada / Suscetibility association between HLA-DRB1 and DQB1 with localized aggressive periodontitis

O objetivo deste estudo foi analisar o perfil HLA classe II em indivíduos portadores de periodontite agressiva localizada e generalizada e comparar com pacientes periodontalmente sadios (grupo controle, GC). As amostras foram compostas por 34 patientes com periodontite agressiva generalizada (média de idade = 29.4, DP ± 4.6), 8 patientes com periodontite agressiva localizada (23.9, DP ± 5.8) e 46 patientes-controle (média de idade 42.8. DP ± 8.6). As amostras foram analisadas pelo método PCR-SSP para análise de genótipo para HLA. As freqüências genéticas dos alelos HLA-DRB1* e HLA-DQB1* foram calculadas. Os resultados mostraram que os alelos HLA-DRB1*08 e -DQB1*04 estavam com sua freqüência significantemente aumentada (p < 0,05) na periodontite agressiva localizada. Sendo assim, podemos concluir que pode existir uma associação de suscetibilidade entre os alelos HLA-DRB1*08 e HLA-DQB1*04, separados ou em combinação, e a periodontite agressiva localizada.