HIV-1C and HIV-1B Tat protein polymorphism in Southern Brazil.

The transactivator of transcription (Tat) is a key HIV regulatory protein. We aimed to identify the frequency of key polymorphisms in HIV-1C compared with HIV-1B Tat protein, chiefly in the cysteine-, arginine-, and glutamine-rich domains and identify novel point mutations in HIV-1B and C sequences from Southern Brazil. This study was the first to investigate the genetic diversity and point mutations within HIV-1 Tat C in a Brazilian cohort. This was an observational, cross-sectional study, which included sequences of HIV-1B (n = 20) and HIV-1C (n = 21) from Southern Brazil. Additionally, 344 HIV-1C sequences were obtained from the Los Alamos database: 29 from Brazil and 315 from Africa, Asia, and Europe. The frequency of C31S substitution on HIV-1 Tat C in Brazil was 82% vs. 10% in the HIV-1B group (p < 0.0001). The frequency of the R57S substitution among the HIV-1C sequences from Brazil was 74% vs. 20% in HIV-1B (p = 0.004), and that of substitution Q63E in HIV-1C was 80% and 20% in HIV-1B (p < 0.0001). The mutation P60Q was more frequent in HIV-1B than in HIV-1C (55% and 6.12%, respectively, p < 0.0001)). Novel point mutations in the HIV-1C and B Tat functional domains were described. The frequency of C31S and other key point mutations in HIV-1 Tat C in Brazil were similar to those described in Africa, although lower than those in India. The Tat-B and C sequences found in Southern Brazil are consistent with biological differences and have potential implications for HIV-1 subtype pathogenesis.

Comparison of Cerebrospinal Fluid Biomarkers for Differential Diagnosis of Acute Bacterial and Viral Meningitis with Atypical Cerebrospinal Fluid Characteristics.

OBJECTIVE: Several cerebrospinal fluid (CSF) biomarkers are used to distinguish between acute bacterial meningitis (BM) and viral meningitis (VM). We compared the ability of lactate and glucose (GL) in CSF and the CSF/blood GL ratio to distinguish between acute BM and VM with typical and atypical CSF characteristics. METHODS: Three hundred and twenty-four CSF reports were included, which were distributed as the acute BM, VM, and normal control groups (n = 63, 139, and 122, respectively). RESULTS: Lactate level in the CSF of acute BM group was 4-fold higher than that in the acute VM and control groups (p < 0.0001). CSF lactate presented higher specificity (92%) and negative predictive value (94%) compared to CSF GL and CSF/blood GL ratio in distinguishing acute BM and VM. Definitive acute BM or VM with atypical CSF cell characteristics was observed in 23.2 and 21.6% of samples, respectively, and these groups showed reduced performance of characteristics of all CSF biomarkers. CSF lactate showed better operational characteristics than those of CSF GL and CSF/blood GL ratio, presenting the highest positive likelihood ratio, and thus aided in the differential diagnosis of VM with atypical CSF. CONCLUSION: The CSF lactate assay can be routinely used in laboratories as a rapid, automated, and easy method that is independent of lactate blood levels.

Influenza Sentinel Surveillance and Severe Acute Respiratory Infection in a Reference Hospital in Southern Brazil

Abstract INTRODUCTION: We report the results of the active surveillance of influenza infections in hospitalized patients and the evaluation of the seasonality and correlation with temperature and rainfall data. METHODS: During the 2-year study period, 775 patients were tested for 15 respiratory viruses (RVs). RESULTS: Most of the 57% of (n=444) virus-positive samples were human rhinovirus and respiratory syncytial virus. However, 10.4% (n=46) were influenza virus (80% FluA; 20% FluB). Age and SARI were significantly associated with influenza. FluB circulation was higher is 2013. CONCLUSIONS: In the post-epidemic period, influenza remains an important cause of hospitalization in SARI patients.

Influenza Sentinel Surveillance and Severe Acute Respiratory Infection in a Reference Hospital in Southern Brazil.

INTRODUCTION: We report the results of the active surveillance of influenza infections in hospitalized patients and the evaluation of the seasonality and correlation with temperature and rainfall data. METHODS: During the 2-year study period, 775 patients were tested for 15 respiratory viruses (RVs). RESULTS: Most of the 57% of (n=444) virus-positive samples were human rhinovirus and respiratory syncytial virus. However, 10.4% (n=46) were influenza virus (80% FluA; 20% FluB). Age and SARI were significantly associated with influenza. FluB circulation was higher is 2013. CONCLUSIONS: In the post-epidemic period, influenza remains an important cause of hospitalization in SARI patients.

Human adenovirus meningoencephalitis: a 3-years' overview.

Human adenovirus (HAdV) has been recognized as a significant viral pathogen implicated in neurological diseases, particularly in immunocompromised patients. However, its involvement in meningoencephalitis remains unclear. The aim of this study was to investigate HAdV and other viral co-infections in the cerebrospinal fluid (CSF) of patients suspected of having either meningoencephalitis or encephalitis. A total of 373 CSF samples from patients under clinical suspicion of neurological viral infection were included in this study. HAdV was investigated by conventional or multiplex real-time PCR, for different time periods. The frequency of HAdV central nervous system (CNS) infection was 1.08%, predominating in female patients with a predisposing condition, and presented with HAdV encephalitis. HAdV CNS infection was found to occur during the months of autumn and winter. The frequency of HAdV detected in CSF positive samples increased after the change in the diagnostic method from conventional to multiplex real-time PCR. There were no specific NMRI or EEG characteristics and two CSF samples with HAdV encephalitis had normal CSF WBC count. There were two cases of co-infection with HIV; no other co-infections with enterovirus or herpes family viruses were detected. All patients had good outcome. Although HAdV is rarely observable in CNS infectious syndromes, it must be investigated particularly in immunocompromised patients.

High frequency of multiple HPV types detection in Fanconi anemia patients oral swabs.

BACKGROUND: Fanconi anemia (FA) is a rare genetic disease usually characterized by bone marrow failure and congenital malformations. The risk of development of malignancies in the oral cavity of FA patients, such as squamous cell carcinoma (SCC), increases significantly after a hematopoietic stem cells transplant (HSCT), and may also be linked with the presence of human papillomavirus (HPV) infections in the oral cavity. We investigated the prevalence and the HPV genotypes in oral mucosa of Brazilian FA patients. METHODS AND RESULTS: Oral swabs of 49 FA patients were collected. The median age of patients was 20 years (range 5-44) and 57% were over 18 years. Oral lesions were present in 20% of all patients, being 90% leukoplakia. HPV DNA was detected in 28% (14/49) of patients, and one of them also reported genital HPV lesions. Sixty-seven percent of all patients had undergone HSCT, including 12 patients (86%) of those with HPV results. Multiple HPV types were detected in 78% and 71% of HPV samples by Sanger sequencing and reverse hybridization methods, respectively. The most prevalent HPV types detected were 6, 11, 18, and 68. CONCLUSIONS: HPV prevalence in the oral mucosa of the assessed FA patients was higher than reported in the general population. Additional studies with collection of sequential samples are needed to know the natural history of the presence of multiple HPV types in these individuals and its association with the development of tumors, to evaluate the implementation of preventive measures, such as vaccination, and to guide early treatment.

Neurological and multiple organ involvement due to Paracoccidioides brasiliensis and HIV co-infection diagnosed at autopsy.

Paracoccidioidomycosis (PCM), caused by Paracoccidioides brasiliensis, is the most prevalent systemic mycosis among immunocompetent patients in Latin America; it is rare in immunocompromised patients. The estimated frequency of central nervous system (CNS) involvement in the HIV/PCM population was 2.5%. We report a case of HIV/P. brasiliensis co-infection, with neurological (NPCM) and multiple organ involvement, indicating a diagnosis of AIDS. PCM diagnosis was established during the autopsy. This is the first described case of HIV/P. brasiliensis co-infection with CNS involvement diagnosed at autopsy. In conclusion, the diagnosis of NPCM is challenging, and it must be considered in the differential diagnosis in HIV-positive patients who reside in or have visited areas in which the condition is endemic and who present with neurological symptoms.

Profile of HIV subtypes in HIV/HBV- and HIV/HCV-coinfected patients in Southern Brazil.

INTRODUCTION:: HIV and viral hepatitis infections are major causes of chronic disease worldwide and have some similarities with regard to routes of transmission, epidemiology, front barriers faced during access of treatment, and strategies for a global public health response. The objective was to describe the HIV-1 subtypes, viral tropism and single-nucleotide polymorphisms (SNPs) of interleukin 28B (IL28B) from a case series of HIV/viral hepatitis coinfected patients from southern Brazil. METHODS:: Clinical and epidemiological data were evaluated by a review of medical records. Periodic blood draws were taken to determine the viral and host characteristics. RESULTS:: This study included 38 patients with HIV/HBV or HIV/HCV coinfection; the median age was 49 years. Thirty-seven (97.4%) were on antiretroviral therapy, 32 (84.2%) had an undetectable viral load, a median CD4+ T-cell count of 452 cells/mm3. HIV-1 subtyping showed 47.4 and 31.6% of patients with subtypes C and B, respectively. Analysis of viral co-receptor usage showed a predominance of the R5 variant (64.7%), with no significant difference between the subtypes. Twenty patients with HIV/HCV coinfection were eligible to receive HCV therapy with pegylated-interferon-alpha plus ribavirin, and 10/20 (50%) of them achieved sustained virological response. SNPs of IL28B were evaluated in 93.3% of patients with HIV/HCV coinfection, and 17 (60.7%) presented the CC genotype. CONCLUSIONS:: In the present case series, a higher frequency of HIV subtype C was found in coinfected patients. However such findings need to be prospectively evaluated with the inclusion of data from regional multicenter analyses.

Profile of HIV subtypes in HIV/HBV- and HIV/HCV-coinfected patients in Southern Brazil

Abstract INTRODUCTION: HIV and viral hepatitis infections are major causes of chronic disease worldwide and have some similarities with regard to routes of transmission, epidemiology, front barriers faced during access of treatment, and strategies for a global public health response. The objective was to describe the HIV-1 subtypes, viral tropism and single-nucleotide polymorphisms (SNPs) of interleukin 28B (IL28B) from a case series of HIV/viral hepatitis coinfected patients from southern Brazil. METHODS: Clinical and epidemiological data were evaluated by a review of medical records. Periodic blood draws were taken to determine the viral and host characteristics. RESULTS: This study included 38 patients with HIV/HBV or HIV/HCV coinfection; the median age was 49 years. Thirty-seven (97.4%) were on antiretroviral therapy, 32 (84.2%) had an undetectable viral load, a median CD4+ T-cell count of 452 cells/mm3. HIV-1 subtyping showed 47.4 and 31.6% of patients with subtypes C and B, respectively. Analysis of viral co-receptor usage showed a predominance of the R5 variant (64.7%), with no significant difference between the subtypes. Twenty patients with HIV/HCV coinfection were eligible to receive HCV therapy with pegylated-interferon-alpha plus ribavirin, and 10/20 (50%) of them achieved sustained virological response. SNPs of IL28B were evaluated in 93.3% of patients with HIV/HCV coinfection, and 17 (60.7%) presented the CC genotype. CONCLUSIONS: In the present case series, a higher frequency of HIV subtype C was found in coinfected patients. However such findings need to be prospectively evaluated with the inclusion of data from regional multicenter analyses.

Reactivation of herpes simplex virus-1 following epilepsy surgery.

PURPOSE: The present study reports a case of encephalitis due to herpes simplex virus-1 (HSV-1), following surgical manipulation of the site of a primary infection. METHODS: Herpes simplex virus-1 infection was confirmed by CSF PCR and DNA sequencing. RESULTS: The patient was an 11-year-old girl who required temporal lobe surgery for epilepsy. She had meningoencephalitis due to HSV at the age of 20 months, and she was treated with acyclovir. Three years later, the patient developed uncontrolled seizures that became more frequent and changed in character at 11 years of age. On the 12th postoperative day, she developed fever and seizures, and she was diagnosed with HSV-1 by positive CSF PCR. She was treated with acyclovir (30 mg/kg/day for 21 days). In this report, we describe the patient and review the relevant literature. CONCLUSION: The authors stress the potential risk of reactivation of HSV encephalitis after intracranial surgery. Herpes simplex virus encephalitis must be considered in neurosurgical patients who develop postoperative seizures and fever.

Viral acute gastroenteritis: clinical and epidemiological features of co-infected patients.

BACKGROUND: Acute gastroenteritis (AGE) is a common disorder that affects children worldwide. It is usually caused by viral agents, including rotavirus, enteric adenovirus, norovirus, and astrovirus groups. Currently, there are few reports about co-infection among these viruses, mainly in Brazil. METHODS: This is a retrospective study in which 84 rotavirus-positive samples from hospitalized patients at a teaching hospital in Southern Brazil, collected in the 2001-2010 period, were analyzed by polymerase chain reaction (PCR) and reverse transcription-polymerase chain reaction (RT-PCR), for the investigation of enteric adenovirus, astrovirus, and norovirus. RESULTS: In total, 12 of the 84 (14%) samples were positive to enteric adenovirus or norovirus. Clinical, laboratory, and demographic data showed statistically significant differences between mono and co-infected patients, including age and depletion rate. CONCLUSIONS: These findings highlight the need for implementation of other enteric virus detection assays in clinical diagnosis for a complete laboratory investigation of hospitalized pediatric patients with AGE, in order to understand the impact of these pathogens on disease severity, spread within hospital, and consequently, prevent the dissemination of nosocomial infections.

Viral acute gastroenteritis: clinical and epidemiological features of co-infected patients

BACKGROUND: Acute gastroenteritis (AGE) is a common disorder that affects children worldwide. It is usually caused by viral agents, including rotavirus, enteric adenovirus, norovirus, and astrovirus groups. Currently, there are few reports about co-infection among these viruses, mainly in Brazil. METHODS: This is a retrospective study in which 84 rotavirus-positive samples from hospitalized patients at a teaching hospital in Southern Brazil, collected in the 2001-2010 period, were analyzed by polymerase chain reaction (PCR) and reverse transcription - polymerase chain reaction (RT-PCR), for the investigation of enteric adenovirus, astrovirus, and norovirus. RESULTS: In total, 12 of the 84 (14%) samples were positive to enteric adenovirus or norovirus. Clinical, laboratory, and demographic data showed statistically significant differences between mono and co-infected patients, including age and depletion rate. CONCLUSIONS: These findings highlight the need for implementation of other enteric virus detection assays in clinical diagnosis for a complete laboratory investigation of hospitalized pediatric patients with AGE, in order to understand the impact of these pathogens on disease severity, spread within hospital, and consequently, prevent the dissemination of nosocomial infections.

Enterovirus and herpesviridae family as etiologic agents of lymphomonocytary meningitis, Southern Brazil.

Viral meningitis is a common infectious disease of the central nervous system (CNS) that occurs worldwide. The aim of this study was to identify the etiologic agent of lymphomonocytary meningitis in Curitiba, PR, Brazil. During the period of July 2005 to December 2006, 460 cerebrospinal fluid (CSF) samples with lymphomonocytary meningitis were analyzed by PCR methodologies. Fifty nine (12.8%) samples were positive. Enteroviruses was present in 49 (83%) samples and herpes virus family in 10 (17%), of these 6 (10%) herpes simplex virus, 1 (2%) Epstein Barr virus, 2 (3%) human herpes virus type 6 and 1 (2%) mixed infection of enterovirus and Epstein Barr virus. As conclusion enterovirus was the most frequent virus, with circulation during summer and was observed with higher frequency between 4 to 17 years of age. PCR methodology is an important method for rapid detection of RNA enterovirus and DNA herpesvirus in CSF.

Enterovirus and herpesviridae family as etiologic agents of lymphomonocytary meningitis, Southern Brazil / Enterovirus e família herpesviridae como agentes etiológicos de meningites linfomonocitárias, no sul do Brasil

Viral meningitis is a common infectious disease of the central nervous system (CNS) that occurs worldwide. The aim of this study was to identify the etiologic agent of lymphomonocytary meningitis in Curitiba, PR, Brazil. During the period of July 2005 to December 2006, 460 cerebrospinal fluid (CSF) samples with lymphomonocytary meningitis were analyzed by PCR methodologies. Fifty nine (12.8 percent) samples were positive. Enteroviruses was present in 49 (83 percent) samples and herpes virus family in 10 (17 percent), of these 6 (10 percent) herpes simplex virus, 1 (2 percent) Epstein Barr virus, 2 (3 percent) human herpes virus type 6 and 1 (2 percent) mixed infection of enterovirus and Epstein Barr virus. As conclusion enterovirus was the most frequent virus, with circulation during summer and was observed with higher frequency between 4 to 17 years of age. PCR methodology is an important method for rapid detection of RNA enterovirus and DNA herpesvirus in CSF.

A meningite viral é uma síndrome infecciosa comum do sistema nervoso central (SNC), que ocorre no mundo inteiro. O objetivo deste estudo foi identificar o agente etiológico de meningite linfomonocitária em Curitiba, PR, Brasil. Durante o período de julho de 2005 a dezembro de 2006, 460 amostras com meningite linfomonocitária foram analisadas por metodologias de PCR. Cinquenta e nove (12,8 por cento) amostras foram positivas. Enterovirus estava presente em 49 (83 por cento) amostras e herpes vírus em 10 (17 por cento), destas 6 (10 por cento) HSV, 1 (2 por cento) EBV, 2 (3 por cento) HHV- 6 e 1 (2 por cento) infecção mista de enterovírus e EBV. Conclui-se que o enterovirus foi o vírus mais frequente, com a circulação durante o verão. Houve maior número de amostras positivas entre 4 a 17 anos. A metodologia de PCR é um importante método para a detecção rápida de RNA de enterovirus e DNA do herpesvirus no LCR.

Quantification of cerebrospinal fluid lactic acid in the differential diagnosis between HIV chronic meningitis and opportunistic meningitis.

BACKGROUND: Approximately 40% of HIV infected patients have chronic meningitis at various stages during the infection, 59% are asymptomatic. This is a diagnosis of exclusion and a confounding factor in cerebrospinal fluid (CSF) analysis, any other causes of chronic meningitis by opportunistic or co-infection must be ruled out. The aim of this study was to analyze CSF lactic acid (LA) as an adjuvant biomarker in chronic meningitis due to HIV. METHODS: CSF LA was quantified in 223 CSF samples by the Dimension AR (Dade Behring, Deerfield, IL, USA), distributed into nine groups: 1) HIV positive with an increase in CSF WBCs (n=17); 2) HIV positive with normal CSF (n=20); 3) enterovirus meningitis (n=33); 4) Herpesviridae meningoencephalitis (n=30); 5) fungal meningitis (n=25); 6) tuberculosis (TB) meningitis (n=17); 7) toxoplasmosis (n=18); 8) neurosyphilis (n=6); 9) control group (n=57). RESULTS: CSF LA (median; IQR) was higher in samples with TB meningitis (5.5; 2.9-7.5 mmol/L) and Cryptococcus neoformans meningitis (3.9; 2.7-5.8 mmol/L) compated with samples with HIV chronic meningitis (1.7; 1.4-1.9 mmol/L) and other groups (p ≤ 0.0001). For the diagnosis of HIV chronic meningitis, using a cut-off of 3.5 mmol/L, CSF LA showed high sensitivity and negative predictive value, although low specificity. CONCLUSIONS: CSF LA helps to discriminate between C. neoformans or TB meningitis and HIV chronic meningitis: CSF LA can be included with the methods currently used to identify these specific pathogens, though it does not replace them. It is rapid, inexpensive and easy to perform, and can be used in developing countries.

Molecular epidemiology of HIV-1 clades in Southern Brazil

Human immunodeficiency virus (HIV) clades B and C account for more than 60 percent of the HIV-1 infections worldwide. In this paper, we describe the profiles of patients infected with subtypes of HIV-1 from the state of Paraná, Southern Brazil, and correlate them with demographic and epidemiological findings. A retrospective analysis of HIV cases reported from 1999-2007 was also performed. Data from 293 patients were reviewed and 245 were older than 13 (58 percent female). The distribution of clades was as follows: B 140 (57 percent), C 67 (23 percent), F 24 (10 percent) and mosaic or unique recombinant forms (URFs) 24 (10 percent). Of the 48 patients younger than 13 years of age (62.5 percent male), vertical transmission occurred in 46 and the distribution of clades was as follows: B 14 (29 percent), C 24 (50 percent), F 7 (15 percent) and URFs 6 (13 percent). There was no significant difference in mortality between HIV-1 subtypes. In both groups, patients infected with clade C tended to have higher rates of injection drug use exposure risk.

Molecular epidemiology of HIV-1 clades in Southern Brazil.

Human immunodeficiency virus (HIV) clades B and C account for more than 60% of the HIV-1 infections worldwide. In this paper, we describe the profiles of patients infected with subtypes of HIV-1 from the state of Paraná, Southern Brazil, and correlate them with demographic and epidemiological findings. A retrospective analysis of HIV cases reported from 1999-2007 was also performed. Data from 293 patients were reviewed and 245 were older than 13 (58% female). The distribution of clades was as follows: B 140 (57%), C 67 (23%), F 24 (10%) and mosaic or unique recombinant forms (URFs) 24 (10%). Of the 48 patients younger than 13 years of age (62.5% male), vertical transmission occurred in 46 and the distribution of clades was as follows: B 14 (29%), C 24 (50%), F 7 (15%) and URFs 6 (13%). There was no significant difference in mortality between HIV-1 subtypes. In both groups, patients infected with clade C tended to have higher rates of injection drug use exposure risk.

Quantitation of cerebrospinal fluid lactic acid in infectious and non-infectious neurological diseases.

BACKGROUND: The differential diagnosis between bacterial and viral meningitis is not easy in some cases. Cerebrospinal fluid (CSF) analysis is essential for establishing this diagnosis. The objectives were to quantitate lactic acid (LA) concentrations in bacterial and viral meningitis, and other central nervous system (CNS) diseases in order to evaluate the diagnostic utility of CSF LA for discriminating bacterial from viral meningitis. METHODS: CSF LA was measured in 139 CSF samples from seven groups: viral meningitis with classic CSF; suspicion of viral meningitis with neutrophils in CSF; bacterial meningitis; non-infectious neurological diseases; chronic meningitis; traumatic lumbar puncture (LP) and normal CSF. RESULTS: CSF LA was higher in bacterial meningitis 8.7 + 5.4 mmol/L compared with viral meningitis (1.9 + 0.6) and the other groups (p < 0.0001). CSF LA in the groups with viral meningitis was not different compared to groups with non-infectious CNS diseases and chronic meningitis. The ability of CSF LA to discriminate bacterial from viral meningitis showed a sensitivity of 80% and specificity of 97%, positive predictive value (PPV) of 94%, and negative predictive value (NPV) of 89%. CONCLUSIONS: CSF LA is a powerful test to discriminate bacterial from viral meningitis with high sensitivity, specificity and predictive values. CSF LA can help in the cases with diagnostic uncertainty.