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Analysis of Microglia Morphology and Number in Wistar Rats Brain After CIDR1α-PfEMP1 Recombinant Protein Injection Erma Sulistyaningsih; Izza Amalia Putri; Sheilla Rachmania
Al-Kauniyah: Jurnal Biologi Vol 18, No 1 (2025): AL-KAUNIYAH JURNAL BIOLOGI
Publisher : Department of Biology, Faculty of Science and Technology, Syarif Hidayatullah State Islami

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15408/kauniyah.v1i1.38242

Abstract

AbstractOne malaria vaccine candidate is Cysteine-rich Interdomain Region 1α (CIDR1α) of Plasmodium falciparumErythrocyte Membrane Protein 1 (PfEMP1), an essential protein involved in the pathogenesis of cerebral malaria. Microglia in the brain act as the first line of defense against brain pathological changes. The study aimed to evaluate the response of brain microglia to the CIDR1α-PfEMP1 recombinant protein injection by observing microglia morphology and number in rat’s cerebral cortex. 12 Wistar rats were divided into the control group, which was injected with normal saline solution, and the treatment group, which was injected with 150 µg CIDR1α-PfEMP1 recombinant protein combined with adjuvants. Injection was conducted thrice within three-week intervals (day 1, 21, and 42). Wistar rats were euthanized on day 56, and histological slides were prepared with Hematoxylin-Eosin staining. Examination using a microscope, 400x, and Fiji Image J software showed microglia morphology of ramified and rod cells in both the control and treatment groups. The microglia number in the control group was 93.00 ± 5.77, and the treatment group was 105.75 ± 15.62. Statistical analysis using an independent t-test showed no significant differences between groups (p= 0.15). The result indicated that the injection of CIDR1α-PfEMP1 recombinant protein did not provoke pathological changes in brain tissue, which induced a microglia response. This study strengthens the potential of the CIDR1α-PfEMP1 recombinant protein as a peptide-based malaria vaccine candidate.AbstrakSalah satu kandidat vaksin malaria adalah Cysteine-rich Interdomain Region 1α (CIDR1α) dari Plasmodium falciparum Erythrocyte Membrane Protein 1 (PfEMP1), protein penting dalam patogenesis malaria serebral. Mikroglia di otak berperan sebagai pertahanan lini pertama terhadap perubahan di otak. Penelitian ini bertujuan mengevaluasi respon mikroglia otak terhadap pemberian protein rekombinan CIDR1α-PfEMP1 dengan mengamati morfologi dan jumlah mikrolia pada korteks serebri otak tikus. 12 tikus Wistar dibagi dalam kelompok kontrol yang diinjeksi normal saline dan  kelompok perlakuan diinjeksi 150 µg protein rekombinan CIDR1α-PfEMP1 yang dikombinasikan dengan adjuvant. Injeksi dilakukan tiga kali dengan interval tiga minggu (hari 1, 21, dan 42). Tikus dieuthanasia pada hari ke-56 dan preparat histologi otak disiapkan dengan pengecatan Hematoxyline-Eosin. Pengamatan menggunakan mikroskop 400x dan Fiji Image J software menunjukkan morfologi ramified dan rod cell pada kelompok kontrol maupun perlakuan. Jumlah mikroglia pada kelompok kontrol 93,00 ± 5,7 sedangkan kelompok perlakuan 105,75 ± 15,62). Analisis statistik menggunakan independent-t test menunjukkan tidak terdapat perbedaan yang bermakna antara 2 kelompok (p= 0,15). Hasil ini mengindikasikan bahwa pemberian protein rekombinan CIDR1α-PfEMP1 tidak menimbulkan patologi pada jaringan otak yang memicu respon mikroglia. Hal ini menguatkan potensi protein rekombinan CIDR1α-PfEMP1 sebagai kandidat vaksin malaria berbasis peptida.
Molecular Method Optimization to Identify Plasmodium falciparum Multidrug Resistance 1 (pfmdr1) gene as a Predictor of Antimalarial Resistance Erma Sulistyaningsih; Rosita Dewi; Sheilla Rachmania; Irawan Fajar Kusuma; Muhammad Rizqi Kholifaturrohmy; Yunita Armiyanti; Sakinatus Sariroh Kholifaturrohmah; Made Prasanti Andriani
Biology, Medicine, & Natural Product Chemistry Vol 14, No 2 (2025)
Publisher : Sunan Kalijaga State Islamic University & Society for Indonesian Biodiversity

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14421/biomedich.2025.142.673-678

Abstract

Several approaches have been designed to control malaria, a disease with high morbidity and mortality, but they face some hurdles. Antimalarial resistance is one of the major challenges for malaria elimination, so the detection of antimalarial resistance is essential. Several molecular markers for antimalarial resistance have been identified, including Plasmodium falciparum multidrug resistance 1 (pfmdr1) gene. This study determined the optimization of molecular techniques to identify the pfmdr1 gene as an antimalarial resistance predictor in Indonesia. The study included patients diagnosed with uncomplicated or severe malaria originating from the health district of Kerom Regency, Papua Province, and Dr. Soebandi Hospital, Jember, East Java Province. Blood samples were collected in the Whatmann filer paper after informed consent. DNA was isolated from dried blood filter paper, and nested PCR was performed using a specific primer, the pfmdr1-A and pfmdr1-B genes. The PCR cycle was optimized based on previous studies. The pfmdr1-A has a similar setting to the earlier study, but the pfmdr1-B had a different optimum setting from the previous study with the annealing temperature of 57oC for nested-1 and 62oC for nested-2. This PCR setting could be used for further examination. The positive results of the amplification indicated the potential for antimalarial resistance in the parasite population. A study on the gene copy number and polymorphism is essential to determine the definitive conclusion on antimalarial resistance.