HAYATI Journal of Biosciences

Isolation and Identification of Vibrio parahaemolyticus Bacteria in Bottlenose Dolphins (Tursiop truncates) in Kendal Conservation Pond, Central Java

Nunak Nafiqoh — 2024-04-22

Aquatic mammals in Indonesia are officially protected. However, there is a lack of research on these mammals, particularly in relation to potential disease-causing bacteria. A study was conducted in a conservation pond to address this gap, where swab samples were taken from the blowholes of aquatic mammals. The focus was on identifying bacteria that could potentially cause infectious diseases in these animals. The results revealed V. parahaemolyticus bacterial isolates, which showed a 98% similarity to the registered V. parahaemolyticus in NCBI. These bacterial isolates exhibited hemolysin properties and demonstrated resistance to trimethoprim, streptomycin, cephalothin, and penicillin antibiotics.

Evaluation of the Virulence Gene Irp2 in Iraqi Patients of Urinary Tract Infections and Other Community-Acquired Illnesses

Shahad Hisham Mahmood — 2024-04-23

A total of sixty-two isolates were tested to assess the presence of the irp2 gene in different isolates of Klebsiella pneumoniae. The isolates discussed in this study were obtained from patients who had acquired illnesses either within the hospital premises or in the surrounding vicinity. These isolates were sourced from three hospitals located in Baghdad, namely Al-Imam Ali, Al-Zaafaranya, and Ibin-Albady hospitals. One interesting thing about Klebsiella pneumoniae is that it makes siderophores, especially yersiniabactin. This is because of a gene that controls this trait. The application of DNA sequencing methodologies has facilitated the identification of the irp2 gene in 44% of Klebsiella pneumoniae. According to amino acid sequences and differentiation of nucleotide, the current work reports findings on the identification of the K. pneumoniae irp2 gene isolates collected from patients in Iraq. This event represents the initial recorded occurrence of such detection. The presence of this gene is considered an unconventional human pathogen. The aim is to explore the correlation between genetic analysis and the diagnosis of genetic variation by examining isolates documented in the global GenBank database (LC791754.1, LC791755.1, LC791756.1, LC791757.1, LC791758.1, LC791759.1, LC791760.1). Additionally, it seeks to provide insights into the magnitude of genetic variation observed within these isolates.

Histological and Molecular Evaluation of the Antiproliferative Activity of Allium ampeloprasum Water Extract Against Oral Mucosa Cell Line (Gingival Cancer)

Maryam Hameed Alwan — 2024-05-07

Gingival carcinoma is a malignant neoplasm affecting the oral mucosa and is associated with significant morbidity and mortality. Allium ampeloprasum var. porrum water extracts have gotten a lot of attention because of their bioactive components, such as polyphenols, flavonoids, and alkaloids, which have a variety of pharmacological activities, including antiproliferative actions. This study aimed to evaluate the histological and molecular effects of Allium ampeloprasum (leek) water extract on the proliferation of the murine gingival cancer cell line. Histological evaluation was conducted to examine morphological changes induced by extract treatment. Molecular mechanisms underlying the observed histological changes were investigated using real-time polymerase chain reaction (PCR). Expression levels of key genes associated with cell proliferation and apoptosis were assessed. Histological findings revealed a dose-dependent decrease (100, 50, 25, 12.5, and 6.25 µg/ml) in cell density and altered cell shape in the treated cell line. Also, the percentage of inhibition for the oral mucosa cell line was high, with a significant P of 0.006, in the treated group compared to the control group. Additionally, water extract has an IC₅₀ value of 61 g/ml. The P53 fold increment of gene expression is 0.6, which means the expression level in the experimental condition is 60% higher than the control. This study provides evidence for the potential antiproliferative activity of Allium ampeloprasum water extract on the oral mucosa cell line. The observed histological changes, coupled with the modulation of key genes involved in proliferation and apoptosis, suggest that leek water extract may have therapeutic implications in managing oral cancer.

Pharmacognostic Evaluation and Antioxidant Activities of Tetracera indica (Christm. and Panz.) Merr

Vera Ladeska — 2024-05-14

The community uses Tetracera indica (Christm. and Panz.) Merr extensively as a traditional medicine. This plant was the main source of raw materials for herbal medicinal products, so identification was needed to ensure its authenticity through pharmacognosy evaluation. The present study evaluated macros- and microscopic characteristics, performed phytochemical analysis, and performed total phenolic (TPC) and flavonoid content (TFC) analysis. The organoleptic and macroscopic studies were carried out through sensory organs. The microscopic characteristics of leaf powder and twigs identification using chloral hydrate and distilled water. Numerous chemical reagents, TLC methods and LC-MS/MS analysis were employed in the phytochemical study. Research employed DPPH and FRAP tests to assess the antioxidant activity. Furthermore, quercetin was used as a marker of antioxidant activity. The morphological marker of the leaf T. indica showed the surface of the leaves is rough bristles, shape elliptic-oblong, margin serratus and phyllotaxy alternate. The stem is lignosus, and the rough stem surface is slightly peeling. The lower leaf surface had paracytic stomata and uniseriate trichomes with tapering ends. Raphide crystals and cork cells were detected in the powder analysis of the twigs. The ethyl acetate extract's twig had the highest TPC and TFC values. Ethyl acetate extract from Tetracera indica twigs has potent antioxidant activity with IC₅₀ = 99.84±0.011 µg/ml (DPPH) and 4296.67±0.024 mol/g (FRAP). The study showed that Tetracera indica contains possible active components that could be developed as novel antioxidants. The result of this study might provide early verification and identification to obtain quality medicinal raw materials.

Antioxidant, Antidiabetic, and Antibacterial Activities of Terminalia bellerica Seed Extracts in Various Solvent Polarities

Atmira Sariwati — 2024-05-14

Terminalia bellerica is well-known for producing edible fruits with pharmacotherapeutic properties. Traditional healers use these species to treat and control diabetes mellitus, its side effects, and other illnesses. Involved in this disease's pathophysiological process, extensive research has been conducted to validate and comprehend these bioactive claims scientifically. This research aims to ascertain the bioactive metabolite contents of different solvent polarities, including ethyl acetate, hexane, distilled water, and methanol. The phenol concentration was determined using the Follin-Ciocalteu procedure to be between 23.45 and 160.41 mg GAE/g. The aluminum chloride colorimetric technique measured flavonoid concentration from 88.52 to 7.12 mg QE/g. The quantitative values of tannic acid, which spanned from 0.78 to 5.32 mg TAE/g, were determined by spectrophotometry UV-VIS. The extracts' capacity to reduce free radical damage ABTS (2, 2' azinobis (3-ethylbenzene-thiazoline-6-sulfonic-acid) and DPPH (2,2-diphenyl-1-picrylhydrazyl) was examined. The extract ethyl acetate exhibited the most significant level of antioxidant activity, with IC₅₀ values for DPPH and ABTS of 28.17 and 22.22 g/ml, respectively. Staphylococcus aureus (23 mm) and were tested for antibacterial and antifungal activity in a methanol extract (21 mm). In vitro, antidiabetic activities were assessed using α-glucoside and α-amylase inhibition. The ethyl acetate extract has α-glucoside inhibition IC₅₀ of 23.04 g/ml and α-amylase inhibition IC₅₀ of 25.35 g/ml. T. bellerica seed includes secondary metabolites that show promise as lead chemicals in creating potent medications.

Characterization of Microplastic Degrading Indigenous Bacteria from Ambon Bay Waters

Idham Halil Zakaria — 2024-05-17

Microplastic degradation by bacteria can degrade low-density polyethylene (LDPE). This study aimed to analyze the potential of Ambon Bay bacteria for microplastic degradation, the condition of microplastics after degradation, and identification of the potential for microplastic degradation. The results of isolation revealed as many as 20 bacterial isolates, which correlated with physicochemical conditions in the waters of Ambon Bay. Nine of them could degrade microplastics as indicated by the presence of a clear zone, namely KA1, KA2, KA3, KA4, KA5, KA9, KA10, KS6, and KS8. They were checked for biofilm formation, microplastic hydrophobicity, and percentage of microplastic weight reduction. Four isolates with the highest percentage of microplastic weight reduction on day 40 were KA1, KA2, KA3, and KA10 at 36.19%, 10.16%, 28.39%, and 17.07%, respectively. The results of LDPE microplastic degradation showed differences using field emission scanning electron microscopy-energy dispersive spectroscopy (FESEM/EDS), attenuated total reflection-fourier transform infrared (ATR-FTIR), and X-ray diffraction (XRD). The bacterial isolates identified were KA1 (Bacillus cereus), KA2 (Bacillus toyonesis), KA3 (Bacillus paramycoides), and KA10 (Escherichia coli). Indigenous bacteria from the waters of Ambon Bay have the potential to degrade LDPE microplastics, which causes structural changes, decreased crystallinity, weight, and C=C groups in microplastics after degradation, with bacterial isolate KA1 identified as Bacillus cereus showing the best potential with degradation of LDPE microplastics by 36.19%.

Black-Pigmented Marine Pseudomonas aeruginosa Exhibiting Anti-Bacterial Activity against Multidrug-Resistant (MDR) Wound Infection Bacteria

Muhammad Evy Prastiyanto — 2024-05-27

The urgency of multidrug-resistant (MDR) bacterial infections in wounds is a significant concern due to the high prevalence of MDR in healthcare settings. Black pigmented marine bacteria, strain PS1C, were isolated from marine sediment on Awur Beach Jepara, Central Java, Indonesia, and cultured in the laboratory. The aim of this research includes molecular identification of strain PS1C, extracting black pigment from strain PS1C, isolation of MDR bacteria from wounds, and evaluating the antibacterial activity of black pigment from strain PS1C against MDR bacteria isolates of wound infections. We examined the 16S rRNA gene sequences strain PS1C to identify the species. Then, the black pigment from PS1C was extracted using methanol: acetone (7:3) solvent. Antibacterial activity was evaluated against MDR bacteria from wounds with the microdilution method. A black-pigmented bacterium was isolated and identified as Pseudomonas aeruginosa PS1C. We found that the black pigment from P. aeruginosa PS1C can be developed as an antibacterial agent against the MDR bacterial isolate of wounds with MIC and MBC values of 6.25-12.5 mg/ml and 6.25-25 mg/ml, respectively. In conclusion, the study's findings highlight the potential of the extract of black pigment from P. aeruginosa PS1C as an antibacterial agent against wound-causing MDR bacteria and reinforce previous research into P. aeruginosa can be isolated from marine sources. Additional in vivo investigations and the identification of the antibacterial activity's mode of action are required.

Cloning, Expression, and Bioinformatics Modeling of Human Papillomavirus Type 52 L1/L2 Chimeric Protein in Escherichia coli BL21 (DE3)

Muh. Chaeril Ikramullah — 2024-05-31

Human papillomavirus (HPV) L1 major capsid protein generates a highly immunogenic virus like particles (VLPs), which have been used as the main component of its prophylactic vaccine. However, the neutralizing antibodies against L1 VLPs are mostly type specific and may not be effective to prevent infection from different strains of HPV. On the other hand, HPV L2 minor capsid protein has low antigenic variation, thus can induce cross-neutralization. This study aims to obtain HPV 52 L1/L2 chimeric protein, which is designed based on HPV type 52 as one of the most circulated high-risk types in Indonesia, to develop a broad-spectrum HPV vaccine. Substitution of HPV 52 H4 helix L1 region with an HPV 52 L2 epitope was carried out using overlap extension PCR. HPV 52 L1/L2 chimeric gene was constructed into pET-SUMO expression vector and expressed in Escherichia coli BL21 (DE3). Bioinformatics modeling suggested that L2 epitope was located inside of the loop region in monomer form, and on the contrary, it was located outside of the pentamer surface. Furthermore, B cell and T cell epitopes predictions were conducted using Immune Epitope Database (IEDB) analysis. The B cell epitopes prediction revealed eleven potential epitopes, whereas the T cell epitopes prediction showed seven potential epitopes for each MHC class I and MHC class II. This study showed that HPV 52 L1/L2 chimeric protein has the potential to induce cross-neutralizing antibodies and can be developed as a promising candidate for a new HPV vaccine.

Phytochemical Screening and Evaluation of Antibacterial, Anticandidal, and Sporicidal Properties of Euphorbia tirucalli Extract in Terengganu, Malaysia

Noor Zarina Abd Wahab — 2024-05-31

Euphorbia tirucalli, commonly known as the pencil cactus or milk bush plant, is used as an alternative medicine. The current study evaluated the phytochemical contents, antibacterial, anticandidal, and antisporicidal potential of the E. tirucalli methanolic extract. The antibacterial and anticandidal activity of E. tirucalli methanolic extract was determined by performing a disc diffusion assay, MIC, MBC, and MCC. The sporicidal activity was tested at different concentrations of extract and exposure times. Phytochemical analyses revealed the presence of alkaloids, saponins, cardiac glycosides, terpenoids, and tannins in E. tirucalli methanolic extract. Results demonstrated inhibition zones of the extract against Gram-positive bacteria were in range of 22.00-7.00 mm. Meanwhile, inhibition zones of the extract against Gram-negative bacteria were in range of 13.00-7.00 mm. All bacteria were inhibited with MIC values at 1.56-25.0 mg/ml and can be completely killed with MBC values of 20-200 mg/ml. Inhibition zones of E. tirucalli methanolic extract against Candida spp. were in the range of 20.00-8.00 mm. All Candida spp. were inhibited with MIC values at 1.56-100.00 mg/ml and MBC values of 100-300 mg/ml. All concentrations of the extract inhibited all Bacillus spp. spores at different exposure times. In conclusion, the methanolic extract of E. tirucalli exhibits antibacterial, anticandidal, and sporicidal activities. The findings indicated that the methanolic extract of E. tirucalli has good potential for prospective nature-based antimicrobial agents.

Distinct Island Lineages of Binturong (Arctictis binturong) from Indonesia and Its Conservation Implications

Andreas Bandang Hardian — 2024-05-31

Binturong (Arctictis binturong) is a threatened carnivore that inhabits the forests of South and Southeast Asia. Despite its wide range, binturong is relatively scarce across its habitat distribution and is currently under the threat of poaching and illegal trade. Captive breeding has unfortunately been conducted rather haphazardly with a lack of origin record maintained, implicating potential risks to the management such as inbreeding or genetic swamping. This study thus aims to characterise the phylogenetic relationship of Indonesian binturong within the context of Southeast Asian binturong and further probe the distinctness of lineages originating from Java, Sumatra, Indonesian Borneo, and Bangka using Cytochrome B (CytB) and Cytochrome C Oxidase Subunit 1 (CO1). Genetic distance, phylogram topology, and haplotype analysis of both encoding genes further corroborate the distinctness of Java, Borneo, and Bangka binturong from other binturong from Indochinese regions such as India, Laos, and Myanmar. Search for prospective single nucleotide polymorphism markers to discriminate island lineages consistently found that each Java, Bangka, and Bornean binturong be distinct from each other and other lineages, especially when assessed using haplotype-based clustering. Assigning binturong originated from Sumatra is nonetheless more complicated, suggesting the possibility. Our findings substantiated the much-needed systematic research of Southeast Asian binturong as ex-situ insurance population management grows in Indonesia and other parts of the world to protect the diversity of binturong lineages and their corresponding unique evolutionary history.

Endoparasites of Wild Javan Gibbon (Hylobates moloch) At Gunung Halimun Salak National Park, Indonesia

Rizka Malintan — 2024-06-12

Infections of endoparasites in primates in natural habitats are highly prevalent and can cause disease, reduce health quality, and disrupt their life. This study investigated endoparasites prevalence value in the endangered Javan gibbon (Hylobates moloch) in Citalahab Forest, Gunung Halimun Salak National Park, Indonesia, from June to August 2022 by collected fecal samples (N = 10) and analyzed it using floatation methods. As a result, we found five genera of nematodes Trichuris trichiura (10% egg worm prevalence, Oesophagostomum spp. (50%), Trichostrongylus spp. (60%), Ancylostoma spp. (80%), and Strongyloides spp. (100%). The prevalence value of the worms in the larvae stage of Trichostrongylus spp. 20% and Strongyloides spp. 70%. Nematode infection status successively is Strongyloides spp., which is, frequently; Trichostrongylus spp. and Ancylostoma spp., which is, often; Oesophagostomum spp. and Trichuris trichiura which is, occasionally. Four species of nematode were found in both ages, and only Trichuris trichiura was found in one adolescent individual. The threat posed by this parasite deserves attention; further research is needed to fill the gap in our knowledge of their pathogenicity and transmission in Javan gibbon.

Variation of Musa spp. in West Kalimantan, Indonesia, Based on rbcL Chloroplast DNA

Ari Sunandar — 2024-06-12

West Kalimantan is home to Musa spp. conservation and genetic assessment of wild banana relatives are important for future breeding purposes. The present study aims to evaluate the genetic relationship of Musa spp. in West Kalimantan by analyzing the rbcL chloroplast DNA using phylogenetic analysis. The methods in this study were sampling, DNA extraction, PCR of rbcL fragment, and data analysis. The specific primer was used to amplify the rbcL chloroplast DNA of ten accessions of Musa spp. in West Kalimantan. The results showed that the area of the rbcL region of Musa spp. in this study was estimated at 587-591 bp. It showed high variability with a conservation level A+T content of 56.95%. The rbcL sequences of Musa spp. have polymorphic sites on 13 numbers of nucleotides. The phylogenetic analysis with an ML algorithm of 35 Musa spp. from West Kalimantan and GenBank data was successfully divided into 4 main clades, and the bootstrap value was 80-81%. This study is expected beneficial for taxonomic, conservation, and banana breeding efforts.

The In Vitro and In Silico Study of α-glucosidase Inhibition by Kombucha Derived from Syzygium polyanthum (Wight) Walp. Leaves

Sitaresmi Yuningtyas — 2024-06-19

Kombucha is a fermented tea drink using a symbiotic culture of bacteria and yeast. This drink has been widely used to maintain blood sugar levels. Meanwhile, leaf boiled water of Syzygium polyanthum (Wight) Walp. has been used as an alternative medicine for diabetes mellitus in Indonesia. If this herb is made into kombucha, it may have higher antihyperglycemic activity than kombucha from tea leaves. However, there are no scientific reports of antihyperglycemic activity from S. polyanthum leaf kombucha by inhibiting alpha-glucosidase. This study aims to determine the activity and kinetics inhibition of S. polyanthum leaves kombucha against α-glucosidase. Samples were prepared at varying concentrations (12.5, 25, 37.5, 50 g/L), while phytochemical components in the products were identified, and the inhibitory activity as well as kinetics were comprehensively analyzed. In silico evaluations were conducted to further explore the inhibitory activity. The results showed that the products contained secondary metabolites such as flavonoids, saponins, and tannins. The inhibitory activity against α-glucosidase ranged from 81.05 to 89.41%. The inhibition mechanism was identified as uncompetitive, with a Michaelis-Menten constant (K_M) of 0.1357 mM and a v_max value of 27.7008 U/ml minute. Several metabolites showed promising inhibition potential due to their strong binding interactions with α-glucosidase, including hydrogen bonding (H-bond), hydrophobic interactions, van der Waals forces, and electrostatic forces. Additionally, two metabolites, farnesol and α-pinene, were found to interact with other human proteins. These observations showed the potential of S. polyanthum leaves kombucha as a health-promoting beverage that might aid blood sugar control in diabetic individuals.

Population Dynamics of Mangrove Clam Pegophysema philippiana (Reeve, 1850) in Davao Region, Southeastern Mindanao, Philippines

Michael Jeriel I. Bersaldo — 2024-06-19

Mangrove clam is the most sought-after bivalve in the Philippines due to its taste, size, and nutrition. Due to its economic importance, this paper aims to determine the population dynamics of mangrove clam P. philippiana harvested in gleaning sites in the Davao region using the FAO ICLARM Stock Assessment Tool (FISAT II). A total of 2493 clams collected from December 2018-December 2020 with sizes ranging from 14-84 mm SL and 2.2-178.6 g were classified according to size classes with 5 mm intervals. The length-weight relationship was computed and showed negative allometry (a = 0.002, b = 2.6205, R²= 0.89). The estimated growth parameters using ELEFAN I was L∞ = 98.64 mm, K = 1.33 year^-1, t₀= -1.07. Length converted catch curve routine estimated Z = 10.27year^-1, M = 1.52 year^-1, F = 8.75 year^-1, E = 0.85, and backward extrapolation generated L_c50= 34.83 mm SL which is below L_m = 65.76 mm SL. Recruitment patterns were highest during July and August 2020, and VPA showed a high F at 60 mm SL. Beverton and Holt Y/R analysis showed E_0.5 = 0.328 and E_max= 0.551, yield isopleths derived from plotted L_c50/L∞ = 0.353 and E = 0.85 values were within quadrant D. The result revealed that L_m>L_c50 implies that mangrove clams gleaned in the area were small and immature. Moreover, F>F_opt and E>E_max and E>E_0.5 suggest that mangrove clam fishery in the region was heavily exploited. The study concluded that the mangrove clam Pegophysema philippiana was overexploited and that a management strategy was needed in the Davao Region, Philippines.

Molecular Detection of Eimeria bovis in Indonesian Beef Cattle Using Nested PCR Technique

Mukh Fajar Nasrulloh — 2024-06-19

Eimeria bovis is a pathogenic protozoan that causes cattle digestive tract infections, which can cause economic losses to farmers. It is necessary to develop specific and accurate detection methods to conserve livestock and prevent coccidiosis in Indonesia. This study aims to detect E. bovis by nested PCR and determine the relationship with reference sequences. A total of 167 samples of beef cattle feces were taken randomly from community farms spread across 18 provinces in Indonesia. The feces were examined natively, and then the oocysts were purified by the sugar flotation method, extracted by KIT extraction, and amplified by the nPCR technique. Positive samples were followed by sequencing and phylogenetic analysis using MEGA 11 software. This study used two pairs of primers (outer and inner) taken from ITS-1 molecular markers. As many as 96 out of 167 samples (57.5%) were positive for Eimeria spp., and 48 of the 96 samples were positive for Eimeria spp. (50%) were detected to be positive for E. bovis based on the presence of a 238 bp DNA fragment. The results of the phylogenetic analysis showed that the study sample formed a separate cluster from the E. bovis cluster from abroad. In conclusion, E. bovis was detected in 16 out of 18 provinces in this study, and the nPCR technique proved to have better sensitivity and specificity.

The Effect of Gel Secretome Hypoxia Mesenchymal Stem Cells to Increase P38 and VEGF Expression in Rats’ Diabetic Wounds

Tarrayuana Rhamadia Hasannuri — 2024-06-20

Mesenchymal stem cells (MSCs) under hypoxic conditions can produce secretomes containing growth factors such as vascular endothelial growth factor (VEGF), accelerating angiogenesis in wound healing disorders in diabetic ulcers. This study aimed to prove the influence of gel secretome MSC hypoxia administration on increasing VEGF and P38 gene expression in rats’ diabetic wounds. An in vivo study was conducted on 25 male Rattus norvegicus, randomly divided into four groups: base gel as a negative control, Gentamycin as a positive control, and gel secretome at a dose of 100 µL, and 200 µL/kg body weight. The differences in P38 and VEGF gene expression were tested using quantitative real-time polymerase chain reaction (qRT-PCR). Wound closure appeared to be fastest in treatment groups at a dose of 100 µL/kg body weight, followed by a dose of 200 µL/kg body weight, followed by Gentamycin and base gel group. The wound closure rate percentage was significantly different in the intervention group compared to the control group (p = 0.000). The results showed a significant difference in P38 and VEGF gene expression between the treatment and control groups (p = 0.000). This study demonstrates the administration of gel secretome hypoxia mesenchymal stem cells increases P38 and VEGF expression in rats’ diabetic wounds.

Anti-SARS-CoV-2 Activity of Andrographis paniculata (Burm.f.) Nees Extract via Inhibition of Spike-mediated Syncytia Formation in HEK293T Cell Model

Pekik Wiji Prasetyaningrum — 2024-06-24

The resurgence of COVID-19 endemic cases at the end of 2023 has underscored the need for effective treatments. Some severe cases of COVID-19 are often characterized by the formation of multinucleated syncytial pneumocytes in the lungs. Therefore, our study aimed to explore the potential of Andrographis paniculata (Burm. f) Nees as an antivirus against SARS-CoV-2, which involves syncitia formation. We utilized the non-toxic concentrations of A. paniculata extract on HEK293T cells determined by MTT assay, which were 1 μg/ml (cell viability 97.96%) and 10 μg/ml (cell viability 95.24%) for further assays. First, we conducted a pseudovirus cellular entry assay as a model of SARS-CoV-2 infection in HEK293T cells expressing hACE2/TMPRSS2. The HEK293T cells were co-transfected with plasmids expressing hACE2 and TMPRSS2, then infected with pseudotyped spike*∆G-GFP rVSV with or without A. paniculata extract. The internalized pseudovirus would trigger GFP expression as a reporter of the infected cells. Next, we performed a syncytia assay by transfecting HEK293T cells with hACE2, TMPRSS2, and SARS-CoV-2 spike expression vectors to induce syncytia formation as a model of intercellular viral transmission. As the results, 10 μg/mL of the extract significantly lowered the number of SARS-CoV-2 pseudovirus-infected cells by 54.69% (P = 0.02) and spike-mediated syncytia formation by 42.39% (P<0.001). In conclusion, our results suggested that A. paniculata has a potential antiviral activity against SARS-CoV-2 by hindering virus infection and cell-to-cell transmission.

In Silico Study, Design, and Expression of an Intranasal Dual Chimeric Vaccine for Indonesian-Based Norovirus GII-2 and Hepatitis B

Ernawati Arifin Giri-Rachman — 2024-07-03

Hepatitis B virus (HBV) remains an important healthcare challenge, leading to liver diseases like cirrhosis and cancer. In response, we created a prophylactic and therapeutic HBV vaccine by integrating HBcAg and HBsAg from HBV genotype B into Norovirus (NoV) GII.2 P domain (PdomGII.2-HBV) for enhanced intranasal delivery. This vaccine also aimed to simultaneously prevent NoV infection, which causes gastroenteritis. Since the selected HBV epitopes have undergone extensive research and are tailored to the Indonesian population, this study focused on identifying NoV epitopes and assessing T cell epitopes coverage of the PdomGII.2-HBV for the Indonesian population. Following that, we expressed the PdomGII.2-HBV protein using Escherichia coli BL21(DE3) and employed a gentle solubilization technique for protein purification. Our in-silico analysis identified two B cell epitopes, along with 15 CD4+T cell epitopes and 35 CD8+T cell epitopes within the GII.2 P domain. These T cell epitopes cover 100% of the Javanese-Sundanese population's HLA allele variations, which constituted the largest demographic group in Indonesia. Subsequently, we successfully purified the presumed PdomGII.2-HBV protein, revealing a molecular weight of 39.5 kDa. Following the successful expression and purification of the presumed PdomGII.2-HBV protein, it is evident that this vaccine design has significant potential, warranting further study.

Genetic Relationship and the Putative Occurrence of A Species Complex Within the Indonesian Calotes (Daudin, 1802) (Squamata, Agamidae) Genus Based on COI Gene Sequences

Muhammad Indra Maulana — 2024-07-05

The Calotes genus presents a challenge due to the complexity of its species. However, research on the cryptic species complex within the Indonesian Calotes genus is still lacking. This study aims to determine the extent of genetic relationships and assess the potential existence of a species complex within the Indonesian genus Calotes (Daudin, 1802) (Squamata, Agamidae) using the partial Cytochrome c Oxidase Subunit 1 (COI) gene sequence as a molecular marker. Samples of the Indonesian Calotes genus in this study were collected from South Lampung (Lampung), Bogor (West Java), and Langkat (North Sumatra). By aligning 582 bp sequence similarities with reference sequences in GenBank, we confirmed that seven out of eight samples analyzed belonged to Calotes vultuosus, while one sample was identified as Calotes versicolor. The identity values ranged from 96 to 100%. The C. vultuosus samples in this study displayed lower genetic distances, ranging from 0 to 3%, with the reference C. vultuosus sequence from Indonesia compared to the reference sequence from India, which ranged from 6 to 9%. Phylogenetic tree reconstruction, utilizing both maximum likelihood with IQ-Tree and Bayesian Inference with BEAST methods, further supports these findings. It reveals distinct groupings between C. vultuosus samples from Indonesia and India. These results suggest the potential occurrence of a species complex within the Indonesian genus Calotes. Furthermore, the inclusion of eight COI gene sequences from two Calotes species in the GenBank database has the potential to confirm the existence of previously undocumented species in Indonesia.

Expression of Immunoglobulin M (IgM) and Immunoglobulin G (IgG) in Normal Wistar Rat Post-Cheral® Administration

Firda Nuri Asyhari — 2024-07-10

Maintaining immunoglobulin levels in the body is important to protect the body from exposure to pathogens. One effort can be made by consuming herbs containing immunomodulatory compounds, such as Cheral^®, which includes a combination of herbs Phyllanthus niruri and Curcuma longa. This research aims to determine the expression of immunoglobulin M (IgM) and immunoglobulin G (IgG) following the administration of Cheral^® to Wistar rats. The study was conducted in vivo, utilizing 24 healthy male Wistar rats for a 90-day treatment period. The research was divided into four treatment groups, including a control group and three dosage groups: Dose 1 (156.25 mg/kg BW), Dose 2 (312.5 mg/kg BW), and Dose 3 (468.75 mg/kg BW). IgM and IgG were isolated from the spleen and analyzed using flow cytometry. Flow cytometry data were analyzed using SPSS with a one-way ANOVA and post hoc test (p-value <0.05). The analysis showed that the relative number of IgM-producing cells in the control group was significantly higher than in the treatment groups, with a difference of 44.40%. In contrast, the relative number of IgG-producing cells in Dose 3 was significantly lower than all other treatment groups, showing a decrease of 29.21%. Overall, the expression of IgG and IgM did not differ substantially across all treatments. The lower IgG and IgM profiles compared to the control group indicate Cheral^®'s ability to prevent infections and maintain the immune system of the rats throughout the treatment period.