Identification of antigenic proteins from salivary glands of female Anopheles maculatus by proteomic analysis

Objective: To identify antigenic proteins from the salivary glands of female Anopheles maculatus using a proteomic approach to find the biomarker candidate for serological tools.Methods: The identification of antigenic proteins of Anopheles maculatus salivary gland used these techniques: one-dimensional gel electrophoresis(sodium dodecyl sulfate polyacrylamide gel electrophoresis), western blot, and liquid chromatography–mass spectrometry.Results: The proteins that have molecular weight(MW) 43 and 34 k Da were the antigenic protein. Computational bioinformatic analysis by Mascot Server revealed seven novel hypothetical proteins(MW: 43 k Da) and two novel hypothetical proteins(MW:34 k Da). Further analysis(BLASTP, antigenicity, epitope mapping, and specificity analysis) showed that two novel proteins were identified as apolipoprotein D and cathepsin D in Anopheles darlingi.Conclusions: The identified proteins are potential to be developed as a biomarker of mosquito bite's exposure.

Clinical features of severe malaria:Protective effect of mixed plasmodial malaria

Objective: To investigate clinically severe malaria patients with Plasmodium falciparum(P. falciparum), Plasmodium vivax(P. vivax) and mixed species infections.Methods: This study was conducted at Dr. Saiful Anwar General Hospital, Malang,Indonesia, from December 2011 to May 2013. Twenty nine patients(mean age of 41 years, 22% female), who suffered from severe malaria according to World Health Organization criteria(major and minor) and other criteria based on previous studies, were selected by consecutive sampling. Blood samples were obtained at admission from peripheral blood for microscopic diagnostic, nested PCR and laboratory examination of blood chemistry. Laboratory results were compared between the groups and correlated to each other.Results: From 29 samples, eight(28%) were diagnosed as P. falciparum mono-infection,12(41%) as P. vivax mono-infection and nine(31%) as mixed infections, confirmed by PCR. Cerebral malaria occurred in P. falciparum or mixed species infection only. Parasitaemia was highest in P. falciparum mono-infection. Mean haemoglobin was significantly lower in P. falciparum than P. vivax infection(P = 0.01). Mean thrombocyte count(77 138/m L) was low in all groups. Mean urea, creatinine, total and direct bilirubin were significantly higher in P. falciparum mono-infection compared to other groups, whereas aspartate aminotransferase and alanine aminotransferase showed no significant differences. Parasitaemia was positively correlated with an increase in urea, creatinine, bilirubin and leucocytosis in all species.Conclusions: Both Plasmodium species can solely or in combination cause severe malaria. Mixed infection was generally more benign than P. falciparum mono-infection and seemed to have some protective effects.

Plasma glutathione and oxidized glutathione level, glutathione/oxidized glutathione ratio, and albumin concentration in complicated and uncomplicated falciparum malaria

Objective: To compare the level of glutathione(GSH) and oxidized glutathione(GSSG),the ratio of GSH/GSSG and the concentration of albumin in plasma of patients with complicated and un-complicated falciparum malaria.Methods: This research was a cross sectional study using comparison analysis with the plasma GSH and GSSG, the ratio of plasma GSH/GSSG and the concentration of plasma albumin as variables. The complicated malaria patients were obtained from Dr. Saiful Anwar Hospital Malang, whereas uncomplicated malaria patients were obtained from the Regency of Pleihari South Kalimantan. Plasma GSH and GSSG levels were determined by the spectrophotometer at the wave length of 412 nm, whereas the concentration of albumin was determined by bromocresol green method in the p H of 4.1.Results: There were no significant differences between the level of plasma GSH and GSSG in complicated and uncomplicated malaria patients, as well as the ratio of plasma GSH/GSSG in the two groups(P = 0.373; P = 0.538; and P = 0.615, respectively, independent ttest). In contrast, the plasma albumin concentration in complicated malaria patients were significantly higher than uncomplicated malaria patients(P = 0.000, Mann Whitney U test).Conclusions: It can be concluded that the average of plasma GSH and GSSG level, also plasma GSH/GSSG ratio in complicated malaria are not different from uncomplicated malaria. Although plasma concentration of albumin in both groups is below the normal range,there is an increase in complicated malaria that might be as compensation of oxidative stress.