Maria A. Santana-Morales

Therapeutic Potential of a Combination of Two Gene-Specific Small Interfering RNAs against Clinical Strains of Acanthamoeba

ABSTRACT Pathogenic strains of the genus Acanthamoeba are causative agents of severe infections, such as fatal encephalitis and a sight-threatening amoebic keratitis. Antimicrobial therapy for these infections is generally empirical, and patient recovery is often problematic, due to the existence of a highly resistant cyst stage in these amoebae. In previous studies, small interfering RNAs (siRNAs) against the catalytic domains of extracellular serine proteases and glycogen phosphorylase from Acanthamoeba were designed and evaluated for future therapeutic use. The silencing of proteases resulted in Acanthamoeba failing to degrade human corneal cells, and silencing of glycogen phosphorylase caused amoebae to be unable to form mature cysts. After the siRNA design and concentration were optimized in order to avoid toxicity problems, cultures of Acanthamoeba were treated with a combination of both siRNAs, and cells were evaluated under an inverted microscope. This siRNA-based treatment dramatically affected the growth rate and cellular survival of the amoebae. These results were observed less than 48 h after the initiation of the treatment. In order to check possible toxic effects of the siRNA combination, three eukaryotic cell lines (HeLa, murine macrophages, and osteosarcoma cells) were treated with the same molecules, and cytotoxicity was examined by measuring lactate dehydrogenase release. The future use of the combination of these siRNAs is proposed as a potential therapeutic approach against pathogenic strains of Acanthamoeba.

Microscopy and molecular biology for the diagnosis and evaluation of malaria in a hospital in a rural area of Ethiopia

BackgroundMalaria is a leading public health problem in Ethiopia. Accurate diagnosis of Plasmodium infections is crucial for the reduction of malaria in tropical areas and for epidemiological studies. The role of light microscopy (LM) as gold standard has been questioned and, therefore, new molecular methods have been developed for the detection of Plasmodium species. The aim of the present work was to compare different malaria diagnostic methods in order to detect the most common species of Plasmodium and to broaden the knowledge of malaria prevalence in a hospital in a rural area in Ethiopia.MethodsA cross-sectional survey of 471 individuals was carried out in a hospital in the rural area of Gambo (Ethiopia). Blood samples were prepared for microscopic observation and collected in filter paper for Seminested-Multiplex PCR (SnM-PCR) and real time PCR (qPCR) testing. The SnM-PCR was considered as the gold standard technique and compared with the rest. Thus, agreement between SnM-PCR and LM was determined by calculating Kappa Statistics and correlation between LM and qPCR quantification was calculated by pair-wise correlation co-efficient.ResultsSamples analysed by LM and SnM-PCR were positive for Plasmodium sp. 5.5% and 10.5%, respectively. Sensitivity was 52.2% by LM and 70% by qPCR. Correlation co-efficient between microscopy counts and qPCR densities for Plasmodium vivax was R2 = 0.586. Prevalence was estimated at 7% (95% CI: 4.7–9.3). Plasmodium vivax was the dominant species detected and the difference was statistically significant (χ2 = 5.121 p < 0.05). The highest prevalence of the parasite (10.9%) was observed in age groups under 15 years old.ConclusionAccurate malaria diagnostic methods have a great effect in the reduction of the number of malaria-infected individuals. SnM-PCR detection of malaria parasites may be a very useful complement to microscopic examination in order to obtain the real prevalence of each Plasmodium species. Although SnM-PCR shows that it is a good tool for the determination of Plasmodium species, today light microscopy remains the only viabletool for malaria diagnosis in developing countries. Therefore, re-inforcement in the training of microscopists is essential for making the correct diagnosis of malaria. Plasmodium vivax was the predominant species in Gambo, a meso-endemic area for this species.

Rural-Urban Differences in Household Treatment-Seeking Behaviour for Suspected Malaria in Children at Bata District, Equatorial Guinea.

Background Malaria remains a major cause of morbidity and mortality among children under five years old in Equatorial Guinea. However, little is known about the community management of malaria and treatment-seeking patterns. We aimed to assess symptoms of children with reported malaria and treatment-seeking behaviour of their caretakers in rural and urban areas in the Bata District. Methodology A cross-sectional study was conducted in the district of Bata and 440 houses were selected from 18 rural villages and 26 urban neighbourhoods. Differences between rural and urban caregivers and children with reported malaria were assessed through the chi-squared test for independence of categorical variables and the t-Student or the non-parametric Mann-Whitney test for normally or not-normally distributed continuous variables, respectively. Results Differences between rural and urban households were observed in caregiver treatment-seeking patterns. Fever was the main symptom associated with malaria in both areas. Malaria was treated first at home, particularly in rural areas. The second step was to seek treatment outside the home, mainly at hospital and Health Centre for rural households and at hospital and private clinic for urban ones. Artemether monotherapy was the antimalarial treatment prescribed most often. Households waited for more than 24 hours before seeking treatment outside and delays were longest in rural areas. The total cost of treatment was higher in urban than in rural areas in Bata. Conclusions The delays in seeking treatment, the type of malaria therapy received and the cost of treatment are the principal problems found in Bata District. Important steps for reducing malaria morbidity and mortality in this area are to provide sufficient supplies of effective antimalarial drugs and to improve malaria treatment skills in households and in both public and private sectors.

Haemoglobin levels for population from Gambo, a rural area of Ethiopia, and their association with anaemia and malaria

BackgroundKnowledge of appropriate reference intervals is critical not only to provide optimal clinical care, but also to enrol populations in medical research. The aim of this study was to generate normal ranges of laboratory values for haemoglobin among healthy Ethiopian adults and children and to determine if anaemia is a possible indicator of malaria in women and children in this area of Ethiopia.MethodsThis study was carried out from January 2008 to May 2010. The reference sample population with malaria-negative consisted of 454 individuals, divided women, men and children. The malaria-infected sample population consisted of 117 individuals. The reference ranges were based on the guidelines from the Clinical and Laboratory Standards Institute. Haemoglobin concentration was determined by Hemo-Control EKF Diagnostic Analyser on whole blood. Testing for malaria-positive and negative infection was done by microscopy and by PCR.ResultsThe lower limits for adult haemoglobin range obtained from this population were slightly higher than those derived from other African populations, but were equal to those established by other studies in Ethiopia and the World Health Organization (WHO). Regarding children, the minimum values were lower than those obtained from different African populations and those established by WHO. The malaria-negative group had anaemia in 35.6% of cases and in the malaria-positive group in 70.9%. There was a stronger, statistically significant association between anaemia and malaria-positive samples than between anaemia and malaria-negative samples in women and both groups of children.ConclusionsThe results from this study are a contribution in the definition of the haemoglobin parameters in African populations, which could be taken as standards for interpretation of laboratory results. The haemoglobin indices in adults from Gambo tended to be higher than other African populations and in children were lower than other studies in Africa. The results also suggest that anaemia is not useful as a supportive diagnostic criterion to monitor and evaluate malaria in women and children from Ethiopia, because a 29.1% of malaria cases will be not detected, because of not having anaemia.

New record of Scaphanocephalus expansus from the Canary Islands (Spain).

SummaryThe osprey, Pandion haliaetus (L), is a cosmopolitan Falconidae that in the Canary Island (Spain) is considered as “endangered” by the Canary Government. No helminthological data of this host are available for this archipelago due its protected status. Forty three Scaphanocephalus expansus were found from the small intestine of one animal. A morphological and morphometric description is given in this work. The species S. expansus can be considered specialists in ospreys, since it has not been found in any other host. It has been found in Europe, Egypt, Asia and North America. According to the references, this is the first report on the helminth fauna of P. haliaetus in the occidental area of Africa.

Prevalence of anemia and associated factors in children living in urban and rural settings from Bata District, Equatorial Guinea, 2013

Anemia in children under 5 years of age is a global public health problem. According to the World Health Organization the current rate of anemia among preschool aged children in Equatorial Guinea is 66%. No information is available above this age. The cross-sectional Prevamal Survey was conducted in 2013 aimed at providing baseline data on malaria prevalence in children aged 2 months-15 years old. Sampling was carried out with the use of a multistage, stratified cluster strategy in the district of Bata, Equatorial Guinea. The χ2 test and adjusted Poisson regression models were applied to assess the association between social-demographic and economic factors, malaria and anemia. A total of 1436 children were tested, out of which 1,421 children (99%) were tested for anemia. Over 85% were anemic; out of them, 284 (24%), 815 (67%) and 111 (9%) children had mild, moderate and severe anemia, respectively. Severe anemia was more frequent among children aged 2–12 months old and those living in rural sites. About 47% tested positive for malaria via a rapid diagnostic test (RDT). This rate was significantly higher in rural villages (66%; p<0.001). The prevalence of anemia and malaria was higher in rural settings (p<0.001). On the other hand, anemia in urban areas displayed a heterogeneity and complexity that differed from the rural environment: in urban neighbourhoods, children with concomitant malaria infection were more likely to be anemic (adjusted prevalence rate (aPR):1.19; CI 95%: 1.12–1.28). Moreover, the prevalence of anemia was higher in children aged above 13 months compared to younger children (p<0.005). Belonging to the poorest wealth tertile were positively (aPR: 1.14, 95% CI: 1.05–1.24) and children’ parents being employees (aPR: 0.86, 95% CI: 0.76–0.96) or self-employed (aPR: 0.86, 95% CI: 0.76–0.97) vs. working in agriculture and/or fishing negatively associated with anemia among urban children. This marked urban-rural variation indicates the importance of targeting specific areas or districts. Strategies aimed at reducing malaria are clearly paramount in this country. Prevention and treatment of other factors associated with the etiology of anemia (e.g., iron deficiency) are also likely necessary to combat the burden of anemia in Equatorial Guinea.

A DEVH-box RNA Helicase from Leishmania braziliensis is Associated to mRNA Cytoplasmic Granules

RNA helicases are ubiquitous enzymes that participate in almost all aspects of RNA processing, including RNA and RNA-protein complex remodelling. In trypanosomatids, which post-transcriptionally regulate gene expression, the formation of different kinds of ribonucleoprotein granules under stress conditions modulates the parasites RNA metabolism. This paper describes the isolation of a putative DEVH-box RNA helicase produced by promastigotes of Leishmania braziliensis. Using a Cy3-labelled dT30 oligo, FISH showed the localization of this protein to mRNA granules under starvation stress conditions. The central region of the protein was shown to be responsible for this behaviour.

Correction: Socio-Economic Differentials in Impoverishment Effects of Out-of-Pocket Health Expenditure in China and India: Evidence from WHO SAGE

There is an error in affiliation 3 for author Santosh Kumar. Affiliation 3 should be: Sam Houston State University, Huntsville, United States of America.