Kemampuan fagositosis makrofag peritoneum mencit yang diimunisasi selama infeksi Plasmodium berghei

ABSTRACT Background: Macrophage represents one of the cellular component of the immune system which plays an important role during malarial infection. Both the number and functional activities including phagocyte activity of these cells increase during the infection. Objectives: This study was carried out to investigate the phagocyte activity of peritoneal macrophages from immunized and non-immunized mice during P. berghei infection. Methods: Swiss mice were divided into two groups, one experimental group was immunized by crude vaccine P. berghei, one control group was not immunized. Phagocyte activity was measured by the ability of mouse peritoneal macrophages to phagocytes latex particles in vitro. Results: In non-immunized mice the percentage of macrophages which were phagocyte latex particles was increased during early infection, reached a peak of about 9 times of the normal level then declined until the mice died. In the immunized mice this activity was increased to reach a peak of about 11 times of the normal level and remained high until recovery. Conclusion: Phagocyte activity of immunized-mice peritoneal macrophages was significantly higher than those of non immunized. The increase of the phagocyte activity seemed to be correlated with the ability of mice to overcome the infection. Key words : Immunization - P. berghei - Effector cells - Macrophages - Phagocytosis Latar belakang penelitian: Makrofag adalah salah satu sel kekebalan yang mempunyai peran penting dalam pertahanan terhadap infeksi malaria. Selain jumlahnya, beberapa aktivitas sel tersebut umumnya meningkat selama infeksi termasuk aktivitas fagositosis. Tujuan penelitian: Penelitian ini dilakukan untuk mengetahui perbedaan kemampuan fagositosis makrofag peritoneum pada mencit yang diimunisasi dan yang tidak diimunisasi selama infeksi P. berghei. Bahan dan cara penelitian: Mencit dibagi menjadi dua kelompok, satu kelompok perlakuan diimunisasi dengan crude vaksin P. berghei sedangkan kelompok kontrol tidak diimunisasi. Kemampuan fagositosis diukur dari kemampuan sel makrofag peritoneum untuk memfagositosis partikel latex in vitro. Hasil penelitian: Pada kelompok mencit yang tidak diimunisasi kemampuan sel makrofag untuk memfagositosis partikel latex meningkat sejak awal infeksi dan pada hari ke-6 mencapai puncak kira-kira 9x dari angka normal, kemudian menurun sampai akhirnya mencit mati pada hari ke-10. Pada mencit yang diimunisasi kemampuan sel untuk memfagositosis partikel latex meningkat sampai 11x pada hari ke-6 kemudian sedikit menurun, tetapi tetap tinggi sampai mencit sembuh

Study on Chloroquine Resistance Transporter (pfcrt) Gene Polymorphism of Plasmodium falciparum in Malaria Patients in Lampung

Introduction: The prevalence of malaria in Lampung Province was increased in the last few years. One of the factors contributes to the increased rate is the widespread of the Plasmodium falciparum resistance to antimalarial drugs. Mutation on the gene encoding Pfcrt protein has been reported to be correlated with this resistance.Objectives: To fi nd out the frequency and distribution of the pfcrt gene polymorphism among Plasmodium falciparum in malaria patients in endemic area at Bandar Lampung and Lampung Selatan, LampungProvince.Methods: Blood samples were collected from malaria falciparum patients in Bandar Lampung and Lampung Selatan by active and passive case detections. Two to three mL of venous blood were collected in tubes with EDTA, and kept in the temperature of -200C before DNA extraction. DNA from each sample was extracted using Guanidine isothiocyanate and Chelex 100 methods. Genes encoding Pfcrt protein were amplifi ed by Nested PCR using TCRP and TCRD primers. The polymorphism of the pfcrt gene was identified by cutting the PCR product using Apo1 restriction enzymes to produce 100bp and 34bp fragments.Results: Forty six samples from P. falciparum-infected patients were collected from the two areas. The genes encoding the Pfcrt protein were succesfully amplifi ed, all 46 PCR products showed 100bp and 34bpfragments after incubation with Apo1 restriction enzyme. It indicated that pfcrt polymorphism was 100%.Conclusion: The frequency of pfcrt gene polymorphisms in patients with malaria falciparum in Bandar Lampung and Lampung Selatan was 100%.Keywords: malaria falciparum, pfcrt, chloroquine, polymorphis

Sekresi Tumor Necrosis Factor dan Reactive Oxygen Intermediates oleh makrofag peritoneum mencit yang distimulasi dengan antigen terlarut Plasmodium falciparum

ABSTRACT Background: Malarial infection is stil one of major health problems in the world. In Indonesia, malarial infection is especially caused by Plasmodium falciparum and Plasmodiun vivax. Host immune responses to malarial infection are complex mechanisms, including the humoral immunity by antibody and cellular immunity by T cells and activated effector cells. Macrophages as an effector cells kill malarial parasite by oxidative and non-oxidative mechanism. Tumor necrosis factor (TNF) and reactive oxygen intermediates (ROI) are mediators produced by macrophages which represent as non-oxidative and oxidative killing respectively. Objectives: Understanding the secretion ability of tumor necrosis factor and reactive oxygen intermediates from soluble antigens of P. falciparum stimulated-peritoneal mouse macrophages. Method: In this study, soluble antigens of P. falciparum stimulated-peritoneal mouse macrophages were tested to produce TNF and ROI in vitro. Secretion of TNF was measured by MTT assay dan ROI by NBT reduction assay. Swiss mice were divided into two groups of 15 mice each. One group was stimulated by soluble antigens as experimental group and the other as control group. Result: Secretion of ,TNF and ROI by soluble antigens of P. falciparum stimulated-peritoneal mouse macrophages were significantly higher (

Prevalence and Risk Factors of Intestinal Protozoan Infection in HIV/AIDS Patients in Dr. Sardjito General Hospital Yogyakarta

Introduction: Intestinal protozoa is one of the etiology of gastroenteritis in developing countries. The risk of intestinal protozoan infection increases among HIV/AIDS patients. HIV/AIDS patients with CD4+ T cell < 200 cells/μL are easily infected by intestinal protozoa causing broad clinical symptoms including diarrhea and even death. However, it can be prevented by understanding various risk factors which have role in thepathogenesis of intestinal protozoan infection.Objectives: To study the prevalence and risk factors which aff ect intestinal protozoan infection among HIV/AIDS patients in RSUP Dr. Sardjito Yogyakarta.Methods: Data from 32 HIV/AIDS patients in RSUP Dr Sardjito Yogyakarta in December 2009-March 2010 were obtained by questionnaires, medical records, and macroscopic-microscopic examination of fecal samples with phormol-eter method and acid-fast staining. Data was analysed using Chi square test and multivariate analysis. A p value less than 0.05 is considered as a signifi cantly diff erent.Results: Prevalence of intestinal protozoan infection in HIV/AIDS patients in RSUP Dr Sardjito Yogyakarta was 81.2%. Intestinal protozoa found in fecal examination were Cryptosporidium sp. (60.98%), Microsporidiumsp. (19.51%), Entamoeba histolytica (9.76%), Cyclospora cayetanensis (4.88%), Blastocystis hominis (2.44%), and Giardia lamblia (2.44%) (n = 26). Bivariate analysis showed that in female HIV/AIDS patients with clinical stadium 1 and 2, CD4+ T cell ≥ 200 cells/μL, had lower risk to be infected by intestinal protozoa (RR = 0.600, 0.065, and 0.026, respectively). Intestinal protozoa were easily found in feces of HIV/AIDS patients with diarrheal symptom. In multivariate analysis, clinical stadium was the most signifi cant factor (Exp(β) = 18.85).Conclusion: Prevalence of intestinal protozoan infection in HIV/AIDS patients in RSUP Dr Sardjito Yogyakarta in December 2009-March 2010 was 81.2%. Clinical stadium with moderate and severe symptoms was the most dominant risk factor for intestinal protozoan infection in HIV/AIDS patients.Keywords: risk factor, intestinal protozoa, CD4+ T cell - HIV/AIDS patien

IN VITRO ANTIPLASMODIAL AND CYTOTOXIC ACTIVITIES OF A SUNGKAI (PERONEMA CANESCENS) LEAF EXTRACT

Objective: The aim of this study was to assess the antiplasmodial and cytotoxic activities and to evaluate the selectivity indices of acetone, ethanol and aqueous extracts of Peronema canescens leaves.Methods: Antiplasmodial activity was measured in vitro against Plasmodium falciparum strains D10 and FCR3 by 72 h incubation at 37 °C in a candle jar. Parasitaemia was calculated by counting the parasite numbers in thin smears. In vitro cytotoxicity was assayed in Vero cells using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and reading the absorbency at 595 nm with an ELISA reader. The assessed parameters included: 50% inhibitory concentration (IC50) of antiplasmodial activity, IC50 of cytotoxic activity and the selectivity index of the Peronema canescens leaf extract.Results: The IC50 values for the acetone, ethanol and aqueous extracts were 26.33±1.65, 37.96±8.17 and 12.26±1.05 μg/ml, respectively, against the Plasmodium falciparum D10 strain and 51.14±8.65, 70.22±14.13 and 34.85±6.04 μg/ml, respectively, against the FCR3 strain. For Vero cells, the IC50 values for the acetone, ethanol and aqueous extracts were 23.37±5.63, 629.46±24.85 and 634.00±144.82 μg/ml, respectively. The selectivity indices of these extracts were 0.89, 16.46 and 51.70, respectively, for the D10 strain and 0.46, 8.90 and 18.00, respectively, for the FCR3 strain.Conclusion: The aqueous extract of Peronema canescens leaves had the highest in vitro antiplasmodial activity and the best selectivity index

Prevalence and risk factors of intestinal protozoan infection among child students with disabilities in Bantul District, Yogyakarta Special Region, Indonesia

Children with disabilities are excluded from many aspects of life. Unfortunately, they have an increased risk of infection from many kinds of pathogens including intestinal protozoan. This study aimed to determine the prevalence of intestinal protozoan infections and to evaluate the associated factors among children with disabilities in Bantul District, Yogyakarta Special Region, Indonesia. A cross-sectional study was conducted at school with special needs between June-December, 2019. A total of 150 participants were recruited through simple random sampling. Stool samples were examined microscopically by formalin-ether concentration and Ziehl-Neelsen staining technique. Age was analyzed using the Mann-Whitney tests, while the other variables used chi-square tests. Multivariable logistic regression was conducted to identify factors associated with intestinal protozoan infections. The adjusted prevalence ratio with a 95% confidence interval at a 5% level of significance was used to measure the strength of association. Overall, there were 15 children infected by intestinal protozoan among 130 subjects with mean age of participants of 9.83 ± 3.1 years. The intestinal protozoan species were Entamoeba histolytica 7 (5.38%), Giardia lamblia 4 (3.08%), Blastocystis hominis 7 (5.38%) and Iodamoeba butschlii 1 (0.77%). Prevalence of intestinal protozoan infection among children with disabilities in Bantul District, Yogyakarta, Special Region was 11.54%. There were no significant correlations between the risk factors and intestinal protozoan infection among children with disabilities (p>0.05)

In vivo Antiplasmodial of the Most Active Fraction and Its Compound of Kapur Leaves (Harmsiopanax aculeatus Harms) Extract Against Plasmodium berghei

Introduction : The rising of Plasmodium resistance towards chloroquine and other antimalarial drugs have encouraged to discover and develop new drugs mainly derived from natural products. Harmsiopanax aculeatus (kapur plant) has traditionally used by people of in Maluku Province to treat malaria.Objectives: The aims of this study were to identify antiplasmodial activity and its chemical constituents of the most active fraction of kapur leaves.Methods: The dried powder of Kapur leaves (1.3 kg) were extracted successively by maceration with n-hexane, ethyl acetate and methanol. After removal the solvents the hexane 15.6 g (1.2%), ethyl acetate 53.3 g (4.1%) and methanol 61.1 g (4.7%) extracts were obtained. Those extracts were assayed for their in vivo antiplasmodial activities by using 4-days suppressive test in Swiss mice infected with Plasmodium berghei, HPIA and identified the compound by GC-MS.Results: The ED50 of hexane, ethyl acetate and methanol extracts were 467.58, 2074.02 and 16.16 mg/kgBW, respectively. Fractionation of the methanol extract gave 18 combined fractions (FG1 – FG18). FG8 was the most active fraction with the IC50 HPIA of 18.22 μg/ml. Phytochemical test of this fraction using spray reagent showed the existence of essential oils, triterpenoids, and phenolic compounds. Separation of FG8using pressed chromatography gave 19 combined fractions (FG8.1-FG8.19). The fraction containing intense blue fluorescent spot (FG8.5) was further separated by PLC fourthly eluted with chloroform. Seven major components with the percentage of compotition more than 3.11% were identified as eugenol (tr = 12.692; 18.22%), isoprophyl myristate (tr = 16.333; 3.99%); bis(2-methylpropyl) phtalat (tr = 16.939; 7.15%); methyl palmitic (tr = 17.442; 3.11%); palmitic acid (tr = 17.883; 25.72%); butyl 2-methylpropyl phtalat (tr = 17.957; 9.37%) and bis(2-ethylhexyl) phtalat (tr = 23.258; 23%).Conclusion: Methanol extract of H. aculeatus was the most potential in vivo antiplasmodial activity. Combined fraction 8 which contain 7 compounds was the most active fraction.Keywords: Harmsiopanax aculeatus Harms, in vivo antiplasmodial, HPIA, PLC, GC-M

A POTENTIAL ZOONOTIC PARASITE: CRYPTOSPORIDIUM PARVUM TRANSMISSION IN RATS, PIGS AND HUMANS IN WEST LOMBOK, INDONESIA.

Background: Cryptosporidium is a neglected zoonotic disease, but with the expansion of human communities its incidence is increasing. Animals such as rats and pigs can act as intermediate hosts and transmit Cryptosporidium to humans due to their proximity. Transmission occurs due to the ability of Cryptosporidium to survive in a new host. The research aimed to identify and describe the transmission of Cryptosporidium from animals to humans. Materials and Methods: This research was a cross sectional study and samples were collected from 84 rats, 205 pigs, and 438 humans in West Lombok. Feces samples were examined using polymerase chain reaction (PCR) and sequencing to isolate the presence of Cryptosporidium, and identify the genetic similarity of the parasites found in rats and pigs with those that infect humans. Results: The PCR results found&nbsp;Cryptosporidium parvum&nbsp;in 4.76% (4/84) in rats; 6.34% (13/205) in pigs; and 0.91% (4/438) in humans. The sequencing results showed genetic kinship of&nbsp;C. parvum&nbsp;in rats, pigs, and humans. There are genetic similarities of Cryptosporidium isolates from West Lombok with&nbsp;C. suis&nbsp;isolates of cattle, from Uganda and&nbsp;C. suis&nbsp;isolates of pigs, from Slovakia. Conclusion: There are genetic similarities of Cryptosporidium in animals and humans, requiring that the Public Health programs in those contaminated areas must receive priority attention to prevent further transmission of these potentially fatal parasites

The Effect of Bacille Calmette-Guerin (BCG) on the Changes in Number and Functional Activities of Mononuclear Phagocytes in Malaria- infected Mice Model

Introduction: Recent studies have indicated that Bacille Calmette-Guerin (BCG) vaccination may have benefi cial eff ect on the survival of infant living in malaria endemic area as well as of malaria-infected mice model. However, the impact of injection of BCG vaccine on the changes in number and functional activities of Mononuclear Phagocytes during malaria-infection in animal model is still poorly understood.Objectives: To evaluate the eff ect of BCG on the changes in number and functional activities of Mononuclear Phagocytes (MPs) during Plasmodium berghei infection.Methods: Two groups of 18 Swiss mice were used. The fi rst group was given 0.1 ml of BCG injection subcutaneously and the second was the control non-BCG group. One week after BCG injection, all mice in both groups were inoculated with 107 Plasmodium berghei infected erythrocytes. The parasitaemia were monitored daily and the number and functional activity of splenic and peritoneal macrophages were tested.Results: The parasites were detected in the blood of both groups on the fi rst day after infection. The parasitaemia in the control group grew slowly until day 3, followed by rapid increased up to 38.96% on day 9. Parasitemia of mouse which still alive on day 12 was 59.6%. The parasitaemia of BCG-injected mice were also increased at lower rate after day 3, and the mice still survive until day 15 after infection. The number of peritoneal macrophages from BCG-injected mice increased to a higher degree compared to the non-BCG injected mice. Moreover, the phagocytic activities of peritoneal macrophages in BCG injected group were increased higher up to twice (200%) of normal levels compared to the non-BCG control group which increase only up to 1,5 times (150%) of the normal levels.Conclusion: The injection of BCG on Plasmodium berghei infected Swiss mice resulted in the extension of survival of the mice until day 15, accompanied by higher increased in number of circulating blood, splenic and peritoneal MPs, and the phagocytic activities of peritoneal MPs up to 137% of the increased in non-BCG mice.Keywords: BCG vaccine, malaria, Plasmodium berghei, macrophages, phagocytosis

The Effect of Pandanus conoideus Lamk Extract to the Serum Level of TNF-α, IL-10 and Parasitemia of Plasmodium berghei Infected in Mice

ABSTRACT Introduction: Study on the effects of red fruit (Pandanusconoideus Lamk) has been conducted with various result.Objectives: In this study, the effect of red fruit extract on the level of cytokines TNF-α, IL-10 and the parasitemia of Plasmodium berghei infected Swiss mice were evaluated.Methods: Quasi-experimental design with pre and post test only control group was applied. Sixty male Swiss mice of 8 weeks old and weighs 20-30 g, was simply randomized into four treatment groups. Group I (K1) was stimulated with the extract for 2 weeks before and 2 weeks after infection with P.berghei. K2 was stimulated with the extract for 2 weeks before infection, K3 was stimulated with the extract for 2 weeks after infection and K4, negative control, was given 0.6% tween 40. The dose of the extract was 7.8 mg/30g mice BW, intra gastric once a day. Serum level of TNF-α and IL-10 was measured by ELISA Sandwich methods and the number of parasitemia were examined microscopically. The difference level of TNF-α, IL-10 and parasitemia of each treatment group were analyzed by t-test, one way anova, honestly significant different (HSD) and multivariate analysis (manova).Results: There were significan cedifferences of parasitemia in K3 group compared to others. Parasitemia on day-3 was 18.464% and reduced to 1.054% on day-9. Parasitemia of K2 group was 13.204% on day-3 and 32.455% on day-9. Parasitemia of negative control group was 27.304% on day-3 and 78.506% on day-9. The TNF-α level of K3 group decreased along with the infection, it was 26.985 pg/Ml on day-0 and 22.244 pg/mL on day-6. The IL-10 level increased at all groups and the highest level was on K3 group.Conclusion: Effect of red fruit extract on P.berghei infected mice was reduced both parasitemia and TNF-α level but increased of IL-10 level.Keywords : Pandanus conoideus Lamk, Malaria, Parasitemia, TNF-α, IL-