Loeki Enggar Fitri

Detection of immunogenic proteins from Anopheles sundaicus salivary glands in the human serum.

INTRODUCTION The saliva of mosquitoes has an important role in the transmission of several diseases, including malaria, and contains substances with vasomodulating and immunomodulating effects to counteract the host physiological mechanisms and enhance pathogen transmission. As immunomodulatory components, salivary gland proteins can induce the generation of specific IgG antibodies in the host, which can be used as specific biomarkers of exposure to Anopheles sundaicus . The objective of this study was to identify immunogenic proteins from the salivary glands of Anopheles sundaicus by reaction with sera from individuals living in malaria-endemic areas who are thus exposed to Anopheles mosquitoes. METHODS IgG antibodies targeting salivary gland proteins in serum samples from individuals living in malaria-endemic areas were measured by enzyme-linked immunosorbent assay (ELISA). Sera from healthy individuals living in non-endemic areas were used as negative controls. Determination of the presence of salivary gland immunogenic proteins was carried out by western blotting. RESULTS Sixteen bands appeared in sodium dodecyl sulfate polyacrylamide gel electrophoresis, with molecule weights ranging from 22 to 144kDa. Among the exposed individuals, IgG responses to salivary gland proteins were variable. Protein bands with molecular weights of 46, 41, 33, and 31kDa were the most immunogenic. These immunogenic proteins were consistently recognized by pooled serum and individual samples from people living in malaria-endemic areas but not by negative controls. CONCLUSIONS These results support the potential use of immunogenic proteins from the salivary glands of Anopheles as candidate markers of bite exposure or in malaria vaccines.

Diversity of the var gene family of Indonesian Plasmodium falciparum isolates.

BackgroundThe large polymorphic protein PfEMP1 is encoded by the var gene family. PfEMP1 has been shown to play an important role as cytoadherence ligand on the surface of infected erythrocytes and thereby contributes to the distinct pathogenesis of malaria. The study explored the diversity of the DBL1α and DBL2β-C2 domains of the protein from Indonesian Plasmodium falciparum field isolates.MethodsSamples of patients with severe and uncomplicated malaria from two different malaria-endemic areas in Indonesia were collected and DNA directly extracted. Dried blood on filter paper was prepared for RNA extraction. PCR amplicons were either cloned and subsequently sequenced or directly sequenced for analysis on nucleotide and amino acid level. Recently published as well as self-designed primers were used for amplification.ResultsBlood from eight patients was finally used for analysis. Seventy-one different sequences out of over 500 DBL1α sequenced clones were observed, resulting in an average of 8.9 different DBL1α sequences per isolate. The average DBL1α sequence similarity within isolates was similar to between isolates. Phylogenetic analysis demonstrated no clustering of sequences regarding strain or geographical origin. The DBL1α sequences were analysed by distribution of semi-conserved features (cysteine/PoLV1-4 grouping) and classified into six sequence groups. The DBL1α cys2 type was observed in all expressed sequences in vivo. Expression of certain DBL sequences implied potential involvement in the pathogenesis. As expected, the DBL2β-C2 domains showed high to moderate homology among each other.ConclusionThe DBL1α domains of PfEMP1 from clinical Indonesian isolates showed high divergence among same isolates and some similarities with other Asia-Pacific strains. Further investigations of important var gene domains with a larger sample size are required to confirm with statistical significance observed associations with severe malaria in Indonesian samples.

Unusual Presentation of Vivax Malaria with Anaemia, Thrombocytopenia, Jaundice, Renal Disturbance, and Melena: A Report from Malang, a Nonendemic Area in Indonesia

Most of the complications of malaria such as anaemia, thrombocytopenia, jaundice, and renal failure are commonly found in Plasmodium falciparum malaria, but the incidence of severe and complicated vivax malaria tends to be increasing. We report two cases of severe Plasmodium vivax malaria from Malang, a nonendemic area in Indonesia. Patients exhibited anaemia, thrombocytopenia, jaundice, renal disturbance, and melena. Microscopic peripheral blood examination and amplification of parasite 18s rRNA by polymerase chain reaction showed the presence of P. vivax and absence of P. falciparum. All patients responded well to antimalarial drugs.

Low Fetal Weight is Directly Caused by Sequestration of Parasites and Indirectly by IL-17 and IL-10 Imbalance in the Placenta of Pregnant Mice with Malaria.

The sequestration of infected erythrocytes in the placenta can activate the syncytiotrophoblast to release cytokines that affect the micro-environment and influence the delivery of nutrients and oxygen to fetus. The high level of IL-10 has been reported in the intervillous space and could prevent the pathological effects. There is still no data of Th17 involvement in the pathogenesis of placental malaria. This study was conducted to reveal the influence of placental IL-17 and IL-10 levels on fetal weights in malaria placenta. Seventeen pregnant BALB/C mice were divided into control (8 pregnant mice) and treatment group (9 pregnant mice infected by Plasmodium berghei). Placental specimens stained with hematoxylin and eosin were examined to determine the level of cytoadherence by counting the infected erythrocytes in the intervillous space of placenta. Levels of IL-17 and IL-10 in the placenta were measured using ELISA. All fetuses were weighed by analytical balance. Statistical analysis using Structural Equation Modeling showed that cytoadherence caused an increased level of placental IL-17 and a decreased level of placental IL-10. Cytoadherence also caused low fetal weight. The increased level of placental IL-17 caused low fetal weight, and interestingly low fetal weight was caused by a decrease of placental IL-10. It can be concluded that low fetal weight in placental malaria is directly caused by sequestration of the parasites and indirectly by the local imbalance of IL-17 and IL-10 levels.

Endoglin Expression and the Level of Tgf-Β Are Increased In the Placental Tissue and Correlated with Low Fetal Weight in Malaria Infected Mice

Ceplukan ( Physalis minima L.) has long been used to treat various conditions in traditional medicine. This study aims to demonstrate the anxiolytic effects of Methanol Extract of Ceplukan Leaves (MECL) in the Elevated Plus Maze (EPM) test and correlate to IL-6 level of ovariectomized rat brain. Total of 24 Wistar rats were divided into six groups: one normal group, one group of 1 month ovariectomized (ovx), one group of 2 months ovx, three groups of 2 months ovx (each given with MECL 500; 1500 and 2500 mg/kg doses for 1 month). The anxiety-like behavior level was measured by EPM test. After EPM test, the brain was removed to measure level of IL-6 by ELISA. The data were processed and analyzed by one-way ANOVA and Pearson correlation. We found that the MECL-treated rats enter the opened-arm higher than the control rats. It indicates that the MECL-treated rats are less anxious than the control rats. The results also show the decreased of IL-6 level in MECL-treated rats.

Strong renal expression of heat shock protein 70, high mobility group box 1, inducible nitric oxide synthase, and nitrotyrosine in mice model of severe malaria

INTRODUCTION Renal damage is a consequence of severe malaria, and is generally caused by sequestration of Plasmodium falciparum -infected erythrocytes in the renal microcirculation, which leads to obstruction, hypoxia, and ischemia. This triggers high mobility group box 1 (HMGB1) to send a danger signal through toll-like receptors 2 and 4. This signal up-regulates inducible nitric oxide (iNOS) and nitrotyrosine to re-perfuse the tissue, and also increases heat shock protein 70 (HSP70) expression. As no study has examined the involvement of intracellular secondary molecules in this setting, the present study compared the renal expressions of HSP70, HMGB1, iNOS, and nitrotyrosine between mice suffered from severe malaria and normal mice. METHODS C57BL/6 mice were divided into an infected group (intraperitoneal injection of 10 6 P. berghei ANKA) and a non-infected group. Renal damage was evaluated using hematoxylin eosin staining, and immunohistochemistry was used to evaluate the expressions of HSP70, HMGB1, iNOS, and nitrotyrosine. RESULTS Significant inter-group differences were observed in the renal expressions of HSP70, HMGB1, and iNOS (p=0.000, Mann-Whitney test), as well as nitrotyrosine (p=0.000, independent t test). The expressions of HSP70 and HMGB1 were strongly correlated (p=0.000, R=1.000). No correlations were observed between iNOS and HMGB, HMGB1 and nitrotyrosine, HSP70 and nitrotyrosine, or iNOS and nitrotyrosine. CONCLUSIONS It appears that HMGB1, HSP70, iNOS, and nitrotyrosine play roles in the renal damage that is observed in mice with severe malaria. Only HSP70 expression is strongly correlated with the expression of HMGB1.

A new endemic area of ovale malaria in Indonesia: A case report from Sumatra Utara Province

Umar Zein, Hadiki Habib, Hadyanto Lim, Loeki Enggar Fitri Dr. Umar Zein Tropical Diseases and Infectious Clinic, Medan, Indonesia Department of Internal Medicine, Faculty of Medicine, Islamic University of Sumatera Utara, Medan, Indonesia Department of Pharmacology, Division of Cardiovascular Medicine, Faculty of Medicine, Methodist University of Indonesia, Medan, Indonesia Department of Parasitology, Faculty of Medicine, Brawijaya University, Malang, Indonesia Asian Pac J Trop Dis 2017; 7(7): 440-441

Pengaruh Artemisin dan Moringa terhadap Derajat Parasitemia, Kadar Malondialdehyde serta Histopatologi Otak Mencit Malaria

Salah satu bentuk infeksi malaria berat adalah malaria serebral ditandai dengan respon inflamasi berlebih dan peningkatan jumlah pembuluh darah di jaringan otak sehingga menyebabkan terbentuknya radikal bebas secara berlebihan. Ekstrak daun Kelor (Moringa oleifera) berpotensi sebagai antioksidan dan antiinflamasi yang diharapkan dapat bekerja sebagai terapi adjuvant untuk kombinasi dengan artemisin. Penelitian ini bertujuan untuk menguji pengaruh kombinasi Artemisin dan ekstrak daun Moringa oleifera terhadap derajat parasitemia, kadar MDA dan gambaran histopatologi pada jaringan otak mencit yang diinfeksi P.berghei. Penelitian ini merupakan True Experimental Design. Sampel penelitian yang digunakan adalah mencit galur BALB/c. Bedasarkan hasil analisis Post Hoc pada hari ke-3 dan ke-7, terdapat perbedaan rerata derajat parasitemia, kadar MDA, dan jumlah pembuluh darah di jaringan otak yang bermakna antara kontrol positif dan semua perlakuan (p<0,05). Pada kelompok yang diterapi Artemisin, didapatkan derajat parasitemia yang lebih rendah dibandingkan dengan kontrol positif (p<0,05), namun lebih tinggi dibandingkan dengan kelompok kombinasi (p<0,05). Rerata kadar MDA turun signifikan pada hari ke 7 terutama pada kelompok kombinasi dosis tertinggi (p=0,000) dibandingkan dengan hari ke 3. Pada pengamatan jumlah pembuluh darah di jaringan otak mencit didapatkan bahwa pada hari ke 7 terjadi penurunan signifikan pada jumlah pembuluh darah di kelompok kombinasi dibandingkan kelompok Artemisin. Dapat disimpulkan kombinasi Artemisin dan ekstrak daun Moringa oleifera lebih baik menurunkan derajat parasitemia, kadar MDA dan jumlah pembuluh darah di jaringan otak mencit dibandingkan dengan terapi artemisin saja. Kata Kunci: Derajat parasitemia, histopatologi otak, malaria, Malondialdehyde

Mid-face location of primary basal cell carcinoma related to cancer aggressivity

Abstract Objective To study the aggressiveness of basal cell carcinoma (BCC) on the mid-face location. Methods A total of 30 patients were diagnosed using specimen biopsy with hematoxylin-eosin stain at Moewardi Public Hospital in Surakarta, Central Java, Indonesia. The age, gender distribution, site of the lesion, as well as clinic-pathological appearance were analyzed. Results There were 30 patients consisting of 46.7% males and 53.3% females with ages ranging from 33 to 91 years old and with the most common occupation, such as farmers (53.6%) and housewives (26.7%). Morpheaform subtypes BCC were more frequent than other types. Based on the predilection, most of the BCC were found to be in the mid-face (76.7%) and using determined criteria of histopathological examination, the aggressive appearance was 77% and non-aggressive BCC was 23%. The BCC on the mid-face location was more aggressive than the other sites ( P Conclusions BCC is the most common skin tumor in humans with rare metastases, which might cause significant damage due to its local recurrences and aggressiveness. BCC on the mid-face is significantly more aggressive than that on the other predilection sites.

Haemozoin Deposits Influence Fetal Weight of Pregnant Mice Infected by Plasmodium berghei

Low birth weight is commonly attributed to malaria in pregnancy, but the cellular and molecular mechanisms that underlie this are incompletely understood. Many of hormones and cytokines are dysregulated in this case and it alters histological structure of placenta which known as placenta malaria. In the placenta malaria, there is an accumulation of infected erythrocytes, macrophages and malarial pigment (haemozoin). This study was conducted to compare the levels of plasma and placenta interferon-gamma (IFN-γ) and haemozoin deposit in pregnant mice that infected by Plasmodium berghei (treatment group) to the normal pregnant mice (control group) and its association with fetal weight. This in vivo experimental laboratory study used pregnant Balb/c mice which divided to control and treatment group. Placentas were staining with Haematoxylin-Eosin (HE) for haemozoin deposits examination. Plasma and placenta levels of IFN-γ examined with ELISA assay. Levels of IFN-γ were higher in plasma than placenta and slightly higher in treatment group than control group, but the differences were not significant (p>0,05). Fetal weight of treatment group was lower than those of control group (p=0,002) however there was no correlation between fetal weight and plasma as well as placenta levels of IFN-γ (p>0,05). Haemozoin deposit was found only in treatment group and influenced weight of fetuses (Spearman=-0,633, p=0,006). Weights of fetuses are more interfered by haemozoin deposit and seemly not by plasma and placenta levels of IFN-γ during malaria infection.Keywords: Fetal weight, gamma interferon, haemozoin, malaria, placenta