THE USE OF ARCHIVED GIEMSA-STAINED BLOOD SMEARS AND RDT FOR PCR-BASED GENOTYPING OF Plasmodium vivax MEROZOITE SURFACE PROTEIN-1 IN CENTRAL KALIMANTAN PROVINCE, INDONESIA: PCR-Based Genotyping of Pvmsp-1 in Central Kalimantan Province, Indonesia

Background: Plasmodium vivax is transmitted most across the country of Indonesia. The country has set out a malaria elimination program by 2030. The information on genetic diversity of malarial parasites relates to malaria transmission in an endemic area may provide the information that can help the malaria control program to achieve the target. Therefore, the purpose of this study was to determine the genetic diversity of the Pvmsp-1 gene in Central Kalimantan Province. Materials and Methods: Samples were 140 of archived Giemsa-stained blood smear and rapid detection test. Samples were divided into the indigenous and migrant populations. After confirmation by single-step PCR, only P. vivax and mixed infection samples were amplified to nested PCR for genotyping of Pvmsp-1 allelic variation in segments F1, F2, and F3. Results: Genotyping of 23 PCR positive samples resulted in 13 genotypes. In segment F1, three allelic variants type A containing subtype A1 (1,050 bp), A2 (350 bp), A3 (150 bp), and type B (100 bp). In segment F2, mono genotypes were detected as variant type A (1,050 bp) and type B3 (150 bp), multiple genotypes were detected as type B containing subtype B1 (250 bp), B2 (200 bp), and B3 (150bp). In segment F3, three allelic variants generated from four mono genotypes were type A (350 bp), type B (300 bp), and two type C (250 bp). Conclusion: The low allelic variation of Pvmsp-1 gene may reflect the actual situation of the low malaria endemic status of the study sites.

THE COMPARISON OF PROPOLIS EFFECTS ON TUMOR NECROSIS FACTOR ALPHA AND MALONDIALDEHYDE BETWEEN INHALATION AND CUTANEOUS ANTHRAX ANIMAL MODELS

Background: Inflammatory response and oxidative stress can be found in anthrax characterized by increased level of serum Tumor Necrosis Factor Alpha (TNF-α) and Malondialdehyde (MDA). The use of antibiotics in anthrax has been known to cause some disturbing side-effects, such as allergic reaction, nausea, vomiting, and antibiotic resistance. Thus, ethanolic extract of propolis (EEP) might be the alternative regimen, due to its anti-inflammatory and antioxidant properties. This study aimed to compare the effects of ethanolic extract of propolis (EEP) on TNF-α and MDA between the inhalation and cutaneous anthrax animal model. Materials and Methods: This was an experimental study with a post-test-only control group design on 40 samples of Rattus norvegicus. Samples were randomized into 5 groups: control, inhalation anthrax model, inhalation anthrax model + EEP, cutaneous anthrax model, and cutaneous anthrax model + EEP. After 14 days, the level of TNF-α and MDA were measured. To compare the data, we used the ANOVA test continued by the post-hoc Turkey test. Results: The results obtained showed that the level of TNF-α and MDA between the inhalation and cutaneous anthrax animal models treated with EEP were statistically different (p < 0.05). The P5 group showed the lowest level of TNF-α (6.822 ± 0.383 pg/ml) and MDA (2.717 ± 0.383 nmol/ml). Conclusion: EEP has a better effect on reducing TNF-α and MDA in cutaneous anthrax animal models compared to the inhalation anthrax animal model.

THE APPLICATION OF CELL-FREE FETAL DNA (cff-DNA) AND SIBLINGS DNA METHODS IN THE PROCESS OF PATERNITY TEST THROUGH CODIS STR LOCI (CSF1PO, THO1, TPOX, AND vWA)

Background: The non-invasive cff-DNA and siblings DNA methods are the latest breakthroughs in the forensic identification process. The use of cff-DNA and siblings DNA as non-invasive techniques in the forensic identification process has, hitherto, not been widely proven. Methods and Materials: This was an analytic observational study. The sample of this study consisted of peripheral blood of women in the second trimester of pregnancy and their two biological children. The kinship analysis was carried out through siblings' DNA and cff-DNA from the mothers through CODIS STR loci (CSF1PO, THO1, TPOX, and vWA). Results: The means of allele sharing between full siblings in loci CSF1PO, THO1, TPOX, and vWA were 0 (13.75%), 1 (44.75%), and 2 (41.50%). The allele sharing found in the study is in line with the one in previous research conducted by Wenk (1998) and the theory proposed by O'Connor (2011), indicating that one allele sharing dominates, contrasting with the finding of previous research conducted by Sosiawan (2020) revealing that 2-allele sharing was more superior. The variation is caused by the ethnicity having a different genetic contribution among the population. The variation can be attributed to historical and demographical processes leading to genetic drift. Conclusion: The mean of SI in 1 allele sharing in CODIS STR loci (CSF1PO, THO1, TPOX, and vWA) has the highest value of 44.5%. The use of cff-DNA of pregnant women as one of the non-invasive techniques can serve as an alternative material in a paternity test.

A FRACTIONAL ORDER HIV/AIDS MODEL USING CAPUTO-FABRIZIO OPERATOR

Background: HIV is a virus that is directed at destroying the human immune system thereby exposing the human body to the risk of been affected by other common illnesses and if it is not treated, it generates a more chronic illness called AIDS. Materials and Methods: In this paper, we employed the fixed-point theory in developing the uniqueness and existence of a solution of fractional order HIV/AIDS model having Caputo-Fabrizio operator. This approach adopted in this work is not conventional when solving biological models by fractional derivatives. Results: The results showed that the model has two equilibrium points namely, disease-free, and endemic equilibrium points, respectively. We showed conditions necessitating the existence of the endemic equilibrium point and showed that the disease-free equilibrium point is locally asymptotically stable. We also tested the stability of our solution using the iterative Laplace transform method on our model which was also shown stable agreeing with the disease-free equilibrium. Conclusions: Numerical simulations of our model showed clear comparison with our analytical results. The numerical solutions show that given fractional operator like the Caputo-Fabrizio operator, it is less noisy and plays a major role in making a precise decision and gives room (‘freedom’) to use data of specific patients as the model can be easily adjusted to accommodate this, as it a better fit for the patients’ data and provide meaningful predictions. Finally, the result showed the advantage of using fractional order derivative in the analysis of the dynamics of HIV/AIDS over the classical case.

A SIMPLE METHOD OF DNA EXTRACTION OF MYCOBACTERIUM TUBERCULOSIS FROM SPUTUM CULTURES FOR SEQUENCING ANALYSIS

Background: Concern has been raised about DNA extraction from Mycobacterium tuberculosis due to its complex procedure. This study demonstrates a simple and fast DNA extraction method of mycobacterial genome to subsequent molecular investigation, such as Polymerization Chain Reaction (PCR) amplification, with species-specific primers and sequencing. Materials and Methods: Total DNA was isolated from M. tuberculosis cultured by using boil method. DNA was evaluated via measures of DNA quantity and quality (absorbance at 230, 260 and 280 nm), DNA integrity (electrophoresis). Molecular tests were tested namely PCR and sequencing. Conclusions: The quality of DNA obtained is acceptable for PCR and sequencing analysis. These findings demonstrate that the method used is inexpensive and suitable for minimum infrastructure facilities.

A CASE REPORT RESPIRATORY MANIFESTATIONS OF COVID-19 STARTING WITH A GASTROINTESTINAL COMPLAINT: A COINCIDENCE OR A CORRELATION?

Background: SARS COV-2 is the cause of the current outbreak of COVID-19. The infection of SARS COV-2 causes changes in the gut- lung axis and the intestinal microbiota pro-inflammatory cytokines interaction which leads to the injury of the gastrointestinal tract. One of the symptoms of COVID-19 outside the respiratory system is a complaint in the GIT. Materials and Methods: We present a COVID-19 case report that begins with a complaint of abdominal pain. Results: There was no previous suspicion of COVID-19, but after a radiological examination and SARS-COV2 positive PCR result, the patient was proven to be suffering from COVID-19. Conclusion: After hospitalization in the ICU for about 14 days, a recovery occurred and the patient was able to go home in a very good clinical condition.

GENERATING RESPONSES IMMUNE IN CELLULAR AND HUMORAL TREATMENT WITH EPITOPE SPIKE, EPITOPE ENVELOPE PROTEIN, AND EPITOPE MEMBRANE PROTEIN SARS-COV-2, HONEY, SAUSSUREA LAPPA, AND NIGELLA SATIVA

Background:Covid-19 has become pandemic in the World, including Indonesia. Our last study showed that HSF could serve as an immunomodulator. Using the exact search, we found that the most immuno-dominant SARS-COV2 epitope, namely A spike protein epitope, B envelope protein epitope, and C membrane protein epitope, we concise to be HF Materials and Methods:We used to post only control design study and mice as an animal model. The research divided mice into four groups, and the first group as control received PBS as a placebo. The second, three, and last four groups gave HF, HSN, and HFHSN (combine HF and HSN). All of the regiment enters the mouth with a special sonde to reach the gastrointestinal organ. We gave HF every week three times and HSN once a day. After administration regiments for a long three weeks, we sacrificed the mice. We evaluated cellular immune responses that are Th-2, Th-17, and NK cells. We check for humoral immune response, TGF-β,IL-17A, IL-4, IgG,IL-4, β-defensin, and s-IgA. Results:Highest profile cellular immunity HF, HSN, and HFHSN were NK cell, Th-2 and Th-17, and the last NK cell, respectively. After that which in humoral immunity, the domination response IgG and IL-4 were HF. But HSN and HFHSN dominated for s-IgA and β-defensin production. By using the study Bio-Informatica, we found HF. Conclusion: If the results of this study are continued to the clinical trial level, it is necessary to recommend additional markers such as CTL (s-IgA and β-defensin in lung tissue)and CPE assay.

N C I D E N C E O F H I G H G R A D E Q T C F P R O L O N G A T I O N A N D I T S M A N A G E M E N T A M O N G P A T I E N T S U N D E R G O I N G T R E A T M E N T F O R D R U G R E S I S T A N T T U B E R C U L O S I S ( D R - T B ) : C A S E S E R I E S .

Background: The World Health Organization (WHO) has approved the use of two new drugs, namely Bedaquiline (Bdq) and Delamanid (Dlm), for treatment of Drug Resistant Tuberculosis (DR-TB). One of the concerns raised with the use of these drugs was QT-interval prolongation. This condition could be serious and life threatening. Hence, knowing the magnitude and its management is very important. This case series identifies the incidence and discusses the management of clinically significant QT-interval prolongation amongst a cohort of patients who have been on these medicines. Materials and Methods: Patients with reports of high grade QT-Interval prolongation (i.e. Grade-3 and Grade-4) were identified from the cohort of 265 patients enrolled on bedaquiline and/or delamanid and discussion is made on the pattern, severity and management of each cases identified. Results: Only 4 (1.5%) out of all 265 patients enrolled on Bedaquiline and/or Delamanid have developed high grade QT-Interval prolongation. And all are managed without permanent discontinuation of both drugs. Conclusion: The Incidence of clinically significant QTcF-interval prolongation among DR-TB patients taking bedaquiline and /or delamanid in Lesotho is low. And almost all cases can be managed with more frequent Electrocardiogram (ECG) monitoring and management of other possible causes of QT-interval prolongation without the need to stop one or both drugs permanently.

EPIDEMIOLOGY OF ZOONOTIC Cryptosporidium spp. ISOLATES IN NJORO SUB_COUNTY NAKURU COUNTY, KENYA

Background: There is no information on human and animal Cryptosporidium spp. in Njoro sub- county. The risk posed to humans and animals within the sub-county is therefore unknown. Materials and Methods: A total of 1476 animal and 378 human fecal samples were evaluated. Multivariate logistic regression was used to evaluate association between infection status and the predisposing factors. Results were expressed as odds ratio (OR) with a 95% confidence interval. Chi-square and Maentel–Haenszel tests were used to quantify relationships among variables. Results: Prevalence of Cryptosporidium spp. was 9.8% in humans, 10.8% in cows, 19.6% in sheep and 4.5% in goats. Prevalence in humans was significantly higher in females 12/37. Infection was highest in the elderly (27.27%), and significantly lower in adolescents and adults at 8.66% and 9.59%, respectively. Goats had lowest overall parasitization at all levels, while sheep had the highest parasitization at levels (+1 and +2). Relatively, humans had the highest parasite counts +3 cases (1.5%). Conclusion: Cryptosporidium spp. is prevalent in Njoro sub-county and domestic animals are important reservoirs and a potential source of zoonosis in humans. Children, elderly and females are at increased risk of infection, especially during rainy season. The study recommends maintenance of proper sanitation when handling domestic animals, treatment of drinking water and use of alternative safer sources of water in order to reduce infection.

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 , 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 . 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 caught in residential areas, 205 pigs, and 438 humans in West Lombok. Fecal 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 Cryptosporidium parvum 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 C. parvum in rats, pigs, and humans. 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.