Hospitalizations of the elderly for reasons related to human immunodeficiency virus (HIV)/acquired immunodeficiency syndrome (AIDS) infection, Rondônia, Brazil, 2010­2018 / Hospitalizações de idosos por motivos relacionados à infecção por vírus da imunodeficiência humana (HIV)/síndrome da imunodeficiência adquirida (AIDS), Rondônia, Brasil, 2010­2018

Introduction: For the elderly, cultural factors and social paradigms predispose to the rejection of condom use, which favors human immunodeficiency virus (HIV) transmission. The North region was the last area for the spread of HIV in Brazil. Rondônia, although with a slight decline in the HIV detection rate in recent years, is still among the states with high mortality rate due to HIV/acquired immunodeficiency syndrome (AIDS) in Brazil. Objective: To describe the demographic and clinical characteristics of the elderly patients who were hospitalized for reasons related to HIV infection in the state of Rondônia, Brazil. Methods: Descriptive study of secondary data recorded from 2010 to 2018. The cause of hospitalization was identified in the field referring to the main and secondary diagnoses of the hospitalization authorization form. Results: Between 2010 and 2018, Rondônia recorded 1,073,932 hospitalizations for any cause in the state's public hospitals. Out of these, 183,803 (17.1%) were of elderly patients over 60 years of age and 249 (0.13%) were due to HIV/AIDS. There was a progressive increase in the number of hospitalizations during the study period, as well as a predominance of younger elderly men (67.1±6.0 years old). The mean length of hospitalizations was 21.3±21.1 days and 57 (24.1%) elderly patients died. Conclusion: Despite all the progress made in HIV prevention and treatment, the state of Rondônia still has a progressive increase in hospitalizations of elderly people for reasons related to HIV/ AIDS. HIV transmission prevention strategies should be emphasized in the elderly population in the state.

Introdução: Fatores culturais e paradigmas sociais predispõem o idoso à rejeição do uso de preservativo, o que favorece a transmissão do vírus da imunodeficiência humana (HIV) nesse grupo. A região Norte foi a última área alvo da disseminação do vírus no Brasil. Rondônia, embora com discreto declínio na taxa de detecção nos últimos anos, encontra-se entre as unidades federativas com mortalidade superior à média nacional. Objetivo: Descrever as características demográficas e clínicas dos idosos que se internaram por motivos relacionados à infecção pelo HIV em hospitais públicos de Rondônia. Métodos: Estudo descritivo de dados secundários registrados no período de 2010 a 2018. A causa da hospitalização foi identificada nos campos referentes aos diagnósticos principal e secundário da autorização de internação hospitalar (AIH). Resultados: Entre 2010 e 2018, o estado de Rondônia registrou 1.073.932 internações por todas as causas nos hospitais públicos/conveniados do SUS. Desse total, 183.803 (17,1%) foram de idosos maiores de 60 anos, das quais apenas 249 (0,13%) foram motivadas por HIV/síndrome da imunodeficiência adquirida (AIDS). Embora de baixa magnitude, observou-se aumento progressivo do número das hospitalizações durante período estudado. Houve um predomínio de internações de idosos do sexo masculino e na sexta década de vida (67,1±6,0 anos). O tempo médio de internação foi de 21,3±21,1 dias e 57 (24,1%) dos idosos evoluíram para óbito. Conclusão: Apesar de todo o progresso obtido na prevenção e tratamento do HIV, o estado de Rondônia ainda apresenta elevação progressiva das hospitalizações de idosos por motivos relacionados ao HIV/AIDS. Estratégias de prevenção da transmissão do HIV devem ser enfatizadas para a população de idosos.

New laboratory perspectives for evaluation of vivax malaria infected patients: a useful tool for infection monitoring.

In recent years, the number of cases with severe Plasmodium vivax malaria has shown an increasing trend. It is, therefore, important to identify routine laboratory markers that best characterize the acute disease phase and can serve as a tool for clinical follow-up of patients. In a cohort study, we followed 87 patients with acute P. vivax monoinfection acquired in an endemic region of the Brazilian Amazon. Forty-two different biochemical and hematological parameters frequently tested in clinical routine were evaluated at the acute phase and the convalescent phase. A total of 42 laboratory tests were performed: biochemical parameters measured were serum lipids levels, aminotransferases, bilirubin, amylase, glucose, urea, creatinine, albumin, globulin, uric acid, C-reactive protein, and alpha-1-acid glycoprotein. Hematological parameters included total and differential white blood cell and platelet counts, hemoglobin concentration, mean platelet volume, platelet width distribution, and plateletcrit. Our results show that several biochemical and hematological parameters were associated with acute phase P. vivax malaria and these parameters reverted to normal values in the convalescent phase. The use of these parameters during diagnosis and follow-up of the infection is a useful clinical tool to evaluate the clinical course and therapeutic response of patients with uncomplicated vivax malaria.

New laboratory perspectives for evaluation of vivax malaria infected patients: a useful tool for infection monitoring

ABSTRACT In recent years, the number of cases with severe Plasmodium vivax malaria has shown an increasing trend. It is, therefore, important to identify routine laboratory markers that best characterize the acute disease phase and can serve as a tool for clinical follow-up of patients. In a cohort study, we followed 87 patients with acute P. vivax monoinfection acquired in an endemic region of the Brazilian Amazon. Forty-two different biochemical and hematological parameters frequently tested in clinical routine were evaluated at the acute phase and the convalescent phase. A total of 42 laboratory tests were performed: biochemical parameters measured were serum lipids levels, aminotransferases, bilirubin, amylase, glucose, urea, creatinine, albumin, globulin, uric acid, C-reactive protein, and alpha-1-acid glycoprotein. Hematological parameters included total and differential white blood cell and platelet counts, hemoglobin concentration, mean platelet volume, platelet width distribution, and plateletcrit. Our results show that several biochemical and hematological parameters were associated with acute phase P. vivax malaria and these parameters reverted to normal values in the convalescent phase. The use of these parameters during diagnosis and follow-up of the infection is a useful clinical tool to evaluate the clinical course and therapeutic response of patients with uncomplicated vivax malaria.

Diagnostic value of neutrophil-to-lymphocyte ratio in patients with leprosy reactions.

INTRODUCTION: Leprosy reactions, classified as type 1 and type 2 reactions, are acute clinical conditions of exacerbation of localized or systemic inflammatory response inpatients with leprosy. No laboratory biomarker is available to predict the emergence of these reactions. Neutrophil-to-lymphocyte ratio (NLR) is an accurate biomarker for diagnosis and prognosis of various inflammatory and neoplastic diseases. OBJECTIVE: This study aimed to investigate the accuracy of the NLR in the diagnosis of leprosy reactions. MATERIALS AND METHODS: NLR was calculated for all patients and a receiver operating characteristic curve (ROC) were generated to identify the NLR cut-off point. RESULTS: A total of 123 patients with leprosy were included, 98 with leprosy reactions of which 56 (45.5%) had type 1 and 42 (34.1%) with type 2. Mean NLR was higher among patients with reactions than among those without. It was also statistically higher among patients with type 2 reactions than in those with type 1 reactions. Receiver operating characteristic curves were generated to identify the NLR cut-off point. The area under the ROC curve was 0.794 for diagnosis of any leprosy reaction and 0.796 for the diagnosis of type 2 reaction. The NLR cut-off points for diagnosis of any leprosy reaction and for type 2 reaction were 2.75 (sensitivity 61.0%, specificity 92.0%, accuracy 77.0%) and 2.95 (sensitivity 81.0%, specificity 74.0%, accuracy 78.0%), respectively. CONCLUSION: These results suggest that NLR could be a potential biomarker for diagnosis of leprosy reaction and useful for discriminating patients with type 2 reactions from those with type 1 leprosy reactions.

Aspartate aminotransferase-to-platelet ratio index (APRI): A potential marker for diagnosis in patients at risk of severe malaria caused by Plasmodium vivax.

Acute infection with Plasmodium vivax, classically associated with benign disease, has been presenting as serious and even fatal disease in recent years. Severe disease is mainly due to biochemical and hematological alterations during the acute phase of infection. In the present cross-sectional study, the aspartate aminotransferase-to-platelet ratio index (APRI) was evaluated as a method for identifying patients at risk of severe vivax malaria. This retrospective study included 130 patients with confirmed P. vivax infection between June 2006 and January 2018. Clinical-epidemiological data were obtained from medical records. Hematological and biochemical parameters were determined using automated equipment. The criteria of severity for infection by Plasmodium falciparum, established by the World Health Organization (WHO), were adapted to classify patients with danger signs of severe vivax malaria. Of the 130 patient's records evaluated, 19 (14.6%) had one or more signs and symptoms of severe malaria. The mean APRI values among patients with and without severe malaria were 2.11 and 1.09, respectively (p = 0.044). Among those with severe disease, the proportion with an APRI value above 1.50 was 30% compared to the 10% among those without severe disease (p = 0.007). The area under the receiver operating characteristic curve (95% CI), calculated to assess the accuracy of the APRI in discriminating between patients with and without severe disease, was 0.645 (0.494; 0.795). An APRI cutoff of 0.74 resulted in sensitivity of 74.0%, specificity of 56.0%, and accuracy of 65.0%. This study shows that the APRI is elevated in patients with evidence of infection by P. vivax. Based on the good sensitivity found in this study, we conclude that this simple index can serve as a diagnostic biomarker to identify patients at risk of severe disease during the acute phase of P. vivax infection.

Decreased uric acid levels in the acute phase of Plasmodium vivax malaria.

INTRODUCTION: Uric acid is one of the compounds associated with the inflammatory process in malaria. It acts as an indicator of cellular damage by activating the immune response and inflammatory process. METHODS: We measured serum concentrations of uric acid in 60 symptomatic patients before and after treatment for malarial infections caused by Plasmodium vivax. RESULTS: Lower serum concentrations of uric acid were found during the acute phase of P. vivax malaria compared to those in its convalescent phase (p < 0.0001). CONCLUSIONS: Patients in the acute phase of malaria had lower uric acid levels than those in its convalescent phase.

Decreased uric acid levels in the acute phase of Plasmodium vivax malaria

Abstract INTRODUCTION Uric acid is one of the compounds associated with the inflammatory process in malaria. It acts as an indicator of cellular damage by activating the immune response and inflammatory process. METHODS: We measured serum concentrations of uric acid in 60 symptomatic patients before and after treatment for malarial infections caused by Plasmodium vivax. RESULTS: Lower serum concentrations of uric acid were found during the acute phase of P. vivax malaria compared to those in its convalescent phase (p < 0.0001). CONCLUSIONS: Patients in the acute phase of malaria had lower uric acid levels than those in its convalescent phase.

Environmental variables and levels of exhaled carbon monoxide and carboxyhemoglobin in elderly people taking exercise.

This article aims to analyze levels of exhaled carbon monoxide, carboxyhemoglobinand cardiopulmonary variables in old people practicing exercise in external environments, and correlate them with climate and pollution factors. Temporal ecological study with118 active elderly people in the city of Cuiabá, in the state of Mato Grosso, Brazil. Data were obtained on use of medication, smoking, anthropometric measurements, spirometry, peak flow, oxygen saturation, heart rate, exhaled carbon monoxide, carboxyhemoglobin, climate, number of farm fires and pollution. Correlations were found between on the one hand environmental temperature, relative humidity of the air and number of farmers' fires, and on the other hand levels of carbon monoxide exhaled and carboxyhemoglobin (p < 0.05).There was a correlation between heart rate and changes in environmental temperature, time of exposure to the sun and relative humidity (p < 0.05). In elderly people, environmental factors influence levels of exhaled carbon monoxide, carboxyhemoglobin and heart rate. There is thus a need for these to be monitored during exercise. The use of a carbon monoxide monitor to evaluate exposure to pollutants is suggested.

Variáveis ambientais e níveis de monóxido de carbono exalado e carboxihemoglobina em idosos praticantes de exercício / Environmental variables and levels of exhaled carbon monoxide and carboxyhemoglobin in elderly people taking exercise

Resumo O objetivo deste artigo é analisar o monóxido de carbono exalado, a carboxihemoglobina e as variáveis cardiopulmonares em idosos praticantes de exercícios em ambientes externos e correlacionar com clima e poluição. Estudo ecológico temporal com 118 idosos ativos em Cuiabá. Foram obtidas informações sobre uso de medicamentos, tabagismo, antropometria, espirometria, peak-flow, saturação de oxigênio, frequência cardíaca, monóxido de carbono exalado, carboxihemoglobina, além de dados climáticos, queimadas e poluição. Temperatura ambiental, umidade relativa do ar e queimadas correlacionaram com monóxido de carbono exalado e carboxihemoglobina (p < 0,05). Frequência cardíaca correlacionou com alterações de temperatura ambiental, tempo de exposição solar e umidade relativa (p < 0,05). Idosos sofrem influências ambientais alterando níveis de monóxido de carbono, carboxihemoglobina e frequência cardíaca, sendo necessário monitoramento destes durante os exercícios, sugerindo-se o uso do monoxímetro para avaliação de exposição a poluentes.

Abstract This article aims to analyze levels of exhaled carbon monoxide, carboxyhemoglobinand cardiopulmonary variables in old people practicing exercise in external environments, and correlate them with climate and pollution factors. Temporal ecological study with118 active elderly people in the city of Cuiabá, in the state of Mato Grosso, Brazil. Data were obtained on use of medication, smoking, anthropometric measurements, spirometry, peak flow, oxygen saturation, heart rate, exhaled carbon monoxide, carboxyhemoglobin, climate, number of farm fires and pollution. Correlations were found between on the one hand environmental temperature, relative humidity of the air and number of farmers’ fires, and on the other hand levels of carbon monoxide exhaled and carboxyhemoglobin (p < 0.05).There was a correlation between heart rate and changes in environmental temperature, time of exposure to the sun and relative humidity (p < 0.05). In elderly people, environmental factors influence levels of exhaled carbon monoxide, carboxyhemoglobin and heart rate. There is thus a need for these to be monitored during exercise. The use of a carbon monoxide monitor to evaluate exposure to pollutants is suggested.

Angiopoietin-2 and angiopoietin-2/angiopoietin-1 ratio as indicators of potential severity of Plasmodium vivax malaria in patients with thrombocytopenia.

INTRODUCTION: Angiogenic factors such as angiopoietin 1 (Ang-1) and angiopoietin 2 (Ang-2) are biomarkers produced during activation and dysfunction of the vascular endothelium in several infectious diseases. The aim of this study was to determine the serum levels of Ang-1 and Ang-2 and to establish their relationship with the main indicators of worst-case prognosis in patients with P. vivax malaria. METHODS: This is a retrospective case-control study nested within a cohort of symptomatic malaria patients. A potentially severe case was defined as a patient that presented at least one of the main indicators of the worst-case prognosis for falciparum malaria, as established by the World Health Organization. Ang-2 and Ang-1 and the Ang-2/Ang-1 ratio were used to analyze the role of angiopoietins as biomarkers in signaling potentially severe vivax malaria. ROC curves were generated to identify a cut-off point discriminating between the angiopoietin concentrations that were most strongly associated with potential infection severity. RESULTS: The serum levels of Ang-2 and the Ang-2/Ang-1 ratio were higher in the case group. In contrast, the serum levels of Ang-1 were lower in the cases than in the control patients. The blood count for platelets showed a positive correlation with Ang-1 and a negative correlation with Ang-2 and with the Ang-2/Ang-1 ratio. The area under the ROC curve (AUC) for serum angiopoietins, as an indicator of worst-case prognosis in a potentially severe P. vivax malarial infection, was larger in the subgroup of patients with platelet counts <75,000/µL. CONCLUSION: This study showed that patients with predictors of worst-case prognoses for P. vivax malaria have lower Ang-1 and higher Ang-2 serum levels (and higher values for the Ang-2/Ang-1 ratio) than controls. Elevated serum levels of Ang-2 and high values for the Ang-2/Ang-1 ratio may potentially be used as predictors of worst-case prognoses for P. vivax malaria, especially in patients with thrombocytopenia.

Factors associated with recurrent Plasmodium vivax malaria in Porto Velho, Rondônia State, Brazil, 2009.

This open retrospective cohort study aimed to describe the incidence of recurrent Plasmodium vivax malaria and associated factors in Porto Velho, Rondônia State, Brazil, in 2009. Data were collected from the National Information System for Malaria Epidemiological Surveillance. There were 23,365 reported P. vivax malaria cases in 2009, 23% of which were classified as relapses. Incidence density of P. vivax recurrence was 45.1/100 patient-years, mostly occurring between the 4th and 13th week after initiating treatment. Male gender, shorter time since onset of symptoms, and higher parasitemia in the initial infection increased the risk of relapse during the year, with a 10% reduction in relative risk for longer symptoms and 11% and 15% increases in relative risk for males and higher initial parasitemia, respectively. However, the results show low clinical relevance for these associations, thereby limiting their applicability to decision-making at the public health level.

Factors associated with recurrent Plasmodium vivax malaria in Porto Velho, Rondônia State, Brazil, 2009 / Fatores associados às recidivas de malária causada por Plasmodium vivax no Município de Porto Velho, Rondônia, Brasil, 2009 / Fatores associados às recidivas de malária causada por Plasmodium vivax no Município de Porto Velho, Rondônia, Brasil, 2009 / Factores asociados a las recidivas de malaria, causada por Plasmodium vivax, en el municipio de Porto Velho, Rondônia, Brasil, 2009

This open retrospective cohort study aimed to describe the incidence of recurrent Plasmodium vivax malaria and associated factors in Porto Velho, Rondônia State, Brazil, in 2009. Data were collected from the National Information System for Malaria Epidemiological Surveillance. There were 23,365 reported P. vivax malaria cases in 2009, 23% of which were classified as relapses. Incidence density of P. vivax recurrence was 45.1/100 patient-years, mostly occurring between the 4th and 13th week after initiating treatment. Male gender, shorter time since onset of symptoms, and higher parasitemia in the initial infection increased the risk of relapse during the year, with a 10% reduction in relative risk for longer symptoms and 11% and 15% increases in relative risk for males and higher initial parasitemia, respectively. However, the results show low clinical relevance for these associations, thereby limiting their applicability to decision-making at the public health level.

Estudo de coorte histórica aberta, construída com informações registradas no Sistema de Informação de Vigilância Epidemiológica da Malária, com objetivo de descrever a incidência e os fatores associados à recidiva de malária causada pelo Plasmodium vivax em Porto Velho, Rondônia, Brasil. Foram notificados 23.365 casos de malária por P. vivax no município, sendo que 23% deles apresentaram recidiva no decorrer de 2009. A densidade de incidência de recidivas foi de 45,1/100 pacientes-ano, ocorrendo, principalmente, entre a 4a e a 13a semana após o início do tratamento. Ser homem, ter menos tempo de sintomas e ter maior nível de parasitemia na infecção inicial aumentaram o risco de recidiva no decorrer do ano, com 10% de redução relativa do risco para maior tempo de sintomas e de 11% e 15% de incremento relativo do risco para o sexo masculino e maiores níveis de parasitemia, respectivamente. Contudo, os resultados demonstram baixa relevância clínica das associações encontradas, comprometendo a sua aplicabilidade na tomada de decisão em nível de saúde pública.

Estudo de coorte histórica aberta, construída com informações registradas no Sistema de Informação de Vigilância Epidemiológica da Malária, com objetivo de descrever a incidência e os fatores associados à recidiva de malária causada pelo Plasmodium vivax em Porto Velho, Rondônia, Brasil. Foram notificados 23.365 casos de malária por P. vivax no município, sendo que 23% deles apresentaram recidiva no decorrer de 2009. A densidade de incidência de recidivas foi de 45,1/100 pacientes-ano, ocorrendo, principalmente, entre a 4a e a 13a semana após o início do tratamento. Ser homem, ter menos tempo de sintomas e ter maior nível de parasitemia na infecção inicial aumentaram o risco de recidiva no decorrer do ano, com 10% de redução relativa do risco para maior tempo de sintomas e de 11% e 15% de incremento relativo do risco para o sexo masculino e maiores níveis de parasitemia, respectivamente. Contudo, os resultados demonstram baixa relevância clínica das associações encontradas, comprometendo a sua aplicabilidade na tomada de decisão em nível de saúde pública. Recidiva; Plasmodium vivax; Malária.

Estudio de cohorte histórica abierta, construida con información registrada en el Sistema de Información de Vigilancia Epidemiológica de la Malaria, con el objetivo de describir la incidencia y los factores asociados a la reaparición de malaria, causada por el Plasmodium vivax, en Porto Velho, Rondônia, Brasil. Se notificaron 23.365 casos de malaria por P. vivax en el municipio, donde un 23% de ellos presentaron recidiva a lo largo del 2009. La densidad de incidencia de recidivas fue de 45,1/100 pacientes-año, produciéndose, principalmente, entre la 4ª y la 13ª semana tras el inicio del tratamiento. Ser hombre, tener menos tiempo síntomas y tener un mayor nivel de parasitemia en la infección inicial aumentaron el riesgo de recidiva a lo largo del año, con un 10% de reducción relativa del riesgo, para un mayor tiempo de síntomas, y de un 11% y 15% de incremento relativo del riesgo para el sexo masculino y mayores niveles de parasitemia, respectivamente. No obstante, los resultados demostraron baja relevancia clínica de las asociaciones encontradas, comprometiendo su aplicabilidad en la toma de decisiones dentro de la salud pública.

Submicroscopic malaria parasite carriage: how reproducible are polymerase chain reaction-based methods?

The polymerase chain reaction (PCR)-based methods for the diagnosis of malaria infection are expected to accurately identify submicroscopic parasite carriers. Although a significant number of PCR protocols have been described, few studies have addressed the performance of PCR amplification in cases of field samples with submicroscopic malaria infection. Here, the reproducibility of two well-established PCR protocols (nested-PCR and real-time PCR for the Plasmodium 18 small subunit rRNA gene) were evaluated in a panel of 34 blood field samples from individuals that are potential reservoirs of malaria infection, but were negative for malaria by optical microscopy. Regardless of the PCR protocol, a large variation between the PCR replicates was observed, leading to alternating positive and negative results in 38% (13 out of 34) of the samples. These findings were quite different from those obtained from the microscopy-positive patients or the unexposed individuals; the diagnosis of these individuals could be confirmed based on the high reproducibility and specificity of the PCR-based protocols. The limitation of PCR amplification was restricted to the field samples with very low levels of parasitaemia because titrations of the DNA templates were able to detect < 3 parasites/µL in the blood. In conclusion, conventional PCR protocols require careful interpretation in cases of submicroscopic malaria infection, as inconsistent and false-negative results can occur.

Sensitivity of nested-PCR for plasmodium detection in pooled whole blood samples and its usefulness to blood donor screening in endemic areas.

Transfusion-transmitted malaria is a severe disease with high fatality rate. Most Brazilian blood banks in the Amazon region perform malaria screening using microscopic examination (thick smears). Since low parasite concentrations are expected in asymptomatic blood donors a high sensitivity test should be used for donor screening. This study determined the sensitivity of a nested-PCR for plasmodium detection in pooled samples. We performed a one-stage criterion validation study with 21 positive samples pooled with samples from ten negative volunteer until three different concentrations were reached (0.33; 0.25; 0.20 parasites/µL - p/µL). Nested PCR was performed as described by Snounou et al. (1993). Sensitivities (and confidence intervals) were determined by stratum of final parasite concentration on the pooled samples. All samples with parasitemia values of 0.33 and 0.25 p/µL had 100% sensitivity (95%CI=86.3-100). One negative result was obtained from a sample with 0.20 p/µL sensitivity=95.2% (95%CI=76.2-99.9). Compared to parasitemia detectable under ideal conditions of thick smear, this nested-PCR in pooled sample was able to detect 40 times more parasites per microliter. Nested-PCR in pooled samples should be considered as a high sensitive alternative to thick smear for donor screening in blood banks at endemic regions. Local authorities need to assess cost:benefit advantages of this method compared to alternatives.

In vitro antimalarial activity of tigecycline against Plasmodium falciparum culture-adapted reference strains and clinical isolates from the Brazilian Amazon.

INTRODUCTION: We evaluated the in vitro antimalarial activity of tigecycline as an alternative drug for the treatment of severe malaria. METHODS: A chloroquine-sensitive Plasmodium falciparum reference strain, a chloroquine-resistant reference strain, and three clinical isolates were tested for in vitro susceptibility to tigecycline. A histidine-rich protein in vitro assay was used to evaluate antimalarial activity. RESULTS: The geometric-mean 50% effective concentration (EC50%) of tigecycline was 535.5 nM (confidence interval (CI): 344.3-726.8). No significant correlation was found between the EC50% of tigecycline and that of any other tested antimalarial drug. CONCLUSIONS: Tigecycline may represent an alternative drug for the treatment of patients with severe malaria.

Submicroscopic malaria parasite carriage: how reproducible are polymerase chain reaction-based methods?

The polymerase chain reaction (PCR)-based methods for the diagnosis of malaria infection are expected to accurately identify submicroscopic parasite carriers. Although a significant number of PCR protocols have been described, few studies have addressed the performance of PCR amplification in cases of field samples with submicroscopic malaria infection. Here, the reproducibility of two well-established PCR protocols (nested-PCR and real-time PCR for the Plasmodium 18 small subunit rRNA gene) were evaluated in a panel of 34 blood field samples from individuals that are potential reservoirs of malaria infection, but were negative for malaria by optical microscopy. Regardless of the PCR protocol, a large variation between the PCR replicates was observed, leading to alternating positive and negative results in 38% (13 out of 34) of the samples. These findings were quite different from those obtained from the microscopy-positive patients or the unexposed individuals; the diagnosis of these individuals could be confirmed based on the high reproducibility and specificity of the PCR-based protocols. The limitation of PCR amplification was restricted to the field samples with very low levels of parasitaemia because titrations of the DNA templates were able to detect < 3 parasites/µL in the blood. In conclusion, conventional PCR protocols require careful interpretation in cases of submicroscopic malaria infection, as inconsistent and false-negative results can occur.

In vitro antimalarial activity of tigecycline against Plasmodium falciparum culture-adapted reference strains and clinical isolates from the Brazilian Amazon

Introduction: We evaluated the in vitro antimalarial activity of tigecycline as an alternative drug for the treatment of severe malaria. Methods: A chloroquine-sensitive Plasmodium falciparum reference strain, a chloroquine-resistant reference strain, and three clinical isolates were tested for in vitro susceptibility to tigecycline. A histidine-rich protein in vitro assay was used to evaluate antimalarial activity. Results: The geometric-mean 50% effective concentration (EC50%) of tigecycline was 535.5 nM (confidence interval (CI): 344.3-726.8). No significant correlation was found between the EC50% of tigecycline and that of any other tested antimalarial drug. Conclusions: Tigecycline may represent an alternative drug for the treatment of patients with severe malaria. .