HAYATI Journal of Biosciences
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Virulence Evaluation of Aeromonas spp. KS-1 Isolated from Kitchen Sponge using Omphisa fuscidentalis Larvae
Get PDFAeromonas spp. causes the human diseases including diarrhea, gastroenteritis, and bacteremia. Aeromonas spp. can be found in kitchen sponge, one of the reservoirs for food-borne bacterial pathogens. Virulence study of Aeromonas spp. in vivo in animal model is important since the animal model can mimic manifestasions in human infections. Omphisa fuscidentalis was chosen for alternative virulence model, since they are in the same taxonomical order with the well-known infections model, Galleria mellonella. Bacterial isolation and selection of kitchen sponge used Brain Heart Infusion agar and Endo Agar, respectively. Bacterial virulence of KS-1 was injected into Omphisa fuscidentalis larvae. Survival percentage and melanization score of infected larvae were evaluated. Hemolymph of larvae with melanization score of 1 and 4 were stained with Giemsa method to observe the hemocyte changes. Bacterial identification of isolate KS-1 based on 16S rRNA sequence resulted in 96.9% identity to Aeromonas spp. strain VS7. Isolate KS-1 injection to O. fuscidentalis revealed higher bacterial dosage resulting more severe symptoms to the larvae according to survival percentage and melanization score. However, statistical analysis showed evaluation of melanization score could distinguish larvae with 106 and 107 CFU/larva dosage injection, while evaluation of survival percentage could not. Hemocyte of larvae with melanization score 1 had larger and more cytoplasmic vacuolization than the score 4 (healthy larvae). Omphisa fuscidentalis is an alternative of insect model for bacterial infections with survival percentage and melanization score as the evaluation. Cytoplasmic vacuolization of hemocyte can be used as larvae’s health indicator in a cellular level
Mangroves in Alas Purwo National Park, Indonesia: Diversity and Its Potential Carbon Services
Get PDFMangroves are important ecosystems that help prevent global warming by storing carbon. A study in Alas Purwo National Park aimed to identify the species diversity and estimate the market value of total carbon from each mangrove species. The study used field sampling and diversity indexes. The study found that the study area has high diversity, with Shannon-Weiner, Margalef, and Pielou\u27s Indexes of H\u27 = 2.276, J = 0.949, and R = 1.453, respectively. Rhizophora apiculata was the dominant species with Above-Ground Carbon (AGC) and Below-Ground Carbon (BGC) stocks of about 34.73 Mg C Ha-1 and an economic value of $1,605, the highest among other species. The results of this study can help improve our understanding of the role of mangrove characteristics for both ecology and the economy
Secondary Metabolite Compounds from Alpinia monopleura Extract and Evaluation of Anti-Inflammatory Activity based on In Vitro and In Silico Studies
Get PDFAlpinia monopleura is one of the endemic plants of Sulawesi, and it has an extensive distribution in the region. Research on chemical compounds and biological activities of A. monopleura is essential to continue as an effort to support the utilization of native plants for medicine. The extract was obtained using the maceration method. The chemical compounds in the extract were identified using Liquid Chromatography Mass Spectrometry (LCMS). Bovine Serum Albumin (BSA) and molecular docking methods were used to evaluate the anti-inflammatory activity. Ten compounds contained in the extract were successfully identified, E-para-coumaric acid (1), trans-ferulaldehyde (2), 3,5,6-trihydroxy-4\u27,7-dimethoxyflavone (3), nevadensin (4), malvalic acid (5), ent-16α,17-hydroxy-19-kauranoic acid (6), 3′,5-dihydroxy-7,4\u27-dimethoxy flavone (7), saurufuran B (8), 5-hydroxy-7,8,2\u27-trimethoxyflavanone (9) and dehydroabietic acid (10). The anti-inflammatory activity of extracts from rhizomes and stems of A. monopleura were 8.62 and 10.59 mg/L, respectively. Some flavonoids (9 and 7) can bind strongly to specific residues around the COX-2 active site, such as Ser530, thereby interfering with the function of the COX-2 enzyme and reducing the production of pro-inflammatory prostaglandins. Thus, A. monopleura extract has the potential to inhibit inflammatory responses through molecular regulation of the COX-2 enzyme
The Effect of Gel Secretome Hypoxia Mesenchymal Stem Cells to Increase P38 and VEGF Expression in Rats’ Diabetic Wounds
Get PDFMesenchymal 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
Black-Pigmented Marine Pseudomonas aeruginosa Exhibiting Anti-Bacterial Activity against Multidrug-Resistant (MDR) Wound Infection Bacteria
Get PDFThe 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\u27s 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\u27s mode of action are required
Extract of Red Okra Pod (Abelmoschus esculentus L. Moench) Chemoprevents N-Methyl-N-Nitrosourea-Induced Kidney Proximal Tubular Cells Damage
Get PDFN-Methyl-N-Nitrosourea (MNU) is a compound that induces oxidative stress resulting in high levels of oxidants and damage to body cells. Red okra contains polyphenol and flavonoid active ingredients as antioxidants. This study aims to analyze the effect of ethanol extract of red okra pods (ROPE) on oxidant levels, antioxidant enzymes, and damage to rat kidney proximal tubule cells due to NMU. This study used 30 rats divided into six treatment groups, namely CN (normal), N (negative control; MNU 50 mg/kg BW), P (positive control; MNU and MTX 50 mg/kg BW), T1 (MNU and ROPE 50 mg/kg BW), T2 (MNU and ROPE 100 mg/kg BW), and T3 (MNU and ROPE 200 mg/kg BW). The treatment was carried out on all groups after eight weeks. The results indicate that malondialdehyde (MDA) and nitrogen oxide (NO) decrease with the ROPE treatment. The glutathione reductase (GSH) activity as an antioxidant enzyme increased T1 and T2, while glutathione peroxidase (GPx) showed an increase in T2 and T3. Furthermore, the biochemical marker of the rat kidney showed lower blood urea nitrogen (BUN) and creatinine (Cre) levels in all treatment groups. Then, the repair of damaged proximal tubule cells showed an increase in normal cells and lower swollen cells; however, there was a degradation in necrotic cells in T2 and T3. It can be indicated that the ROPE can act as an antioxidant that can reduce MDA and NO levels, increase GSH and GPx levels, and reduce damage to proximal renal tubule cells due to MNU
Histological and Molecular Evaluation of the Antiproliferative Activity of Allium ampeloprasum Water Extract Against Oral Mucosa Cell Line (Gingival Cancer)
Get PDFGingival 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
Characterisation of ACE2-like Enzyme from Bacillus cereus sp. as an Alternative Treatment for COVID-19 Patients
Get PDFSARS-CoV-2 infection can lead to downregulation of ACE2, raising pro-inflammatory Angiotensin II levels. Recombinant human ACE2 protein therapy can restore homeostasis but is costly. An alternative is an ACE2-like enzyme with similar effects. The previous research identified a carboxypeptidase protein from Bacillus cereus sp. (rBceCP) as a potential ACE2-like enzyme, but it has not been characterized well. This study characterizes, expresses, and tests the activity of the rBceCP). rBceCP structure and properties were predicted using Prabi and Expasy. The in vitro approach included protein expression optimization, hydrolysis tests, and inhibition tests. In silico analysis revealed the protein is a homodimer with 53.66% α-helix and a molecular weight of 58.99 kDa. The protein is stable, hydrophilic, and has an isoelectric point at pH 4.93, indicating it can be expressed using the E. coli system. Expression of rBceCP showed no significant differences across IPTG concentrations (p-value >0.05). The protein hydrolysis activity of rBceCP was similar to the control, though purified samples had lower activity than crude samples. Inhibition activity in crude and purified samples showed no significant differences (p-value >0.05) and was higher than the control. Thus, rBceCP has potential as an ACE2-like enzyme and a therapeutic candidate for COVID-19
In Silico and In Vitro Inhibitory Activity of Indonesian Herbal Compound Extracts against SARS-COV-2 Recombinant Papain-Like Protease
Get PDFThe SARS-CoV-2 papain-like protease (PLpro) is essential for viral replication and a promising target for drug discovery. This study explored the inhibitory potential of compounds from Indonesia herbals Butterfly pea flower (Clitoria ternatea L), Star fruit leaves (Averrhoa carambola L.), and Java plum leaves (Syzygium cumini (L.) Skeels) against PL pro through molecular docking and in vitro assays. The molecular docking method utilized the target protein PLpro (PDB ID: 7CMD), with the native ligand obtained from compounds identified in these plant extracts. The compounds were identified using the KNApSAcK database and analyzed for drug-likeness based on Lipinski\u27s Rule of Five. The physicochemical characteristics affecting absorption, distribution, metabolism, excretion, and toxicity (ADMET) were determined using the pkCSM descriptor algorithm protocol. Validation was performed using the redocking method, achieving an RMSD score of 0.728 Å, which indicated validity (RMSD <2.0 Å). The results identified four ligands with the lowest binding affinities from these extracts: (-)-Epicatechin 3-O-gallate, folic acid, petunidin 3-glucoside, and ellagic acid, with binding scores of -8.6, -8.3, -7.1, and -7.1 kcal/mol, respectively. Prior to conducting the PLpro in vitro inhibition assay, a fluorescence-based inhibition assay was performed using Z-RLRGG-AMC as the substrate and GRL0617as the control inhibitor. All extracts were subjected to 70% ethanol maceration. The IC50 value of GRL0617 was 3.38 μM, while fluorescence tests showed that Java plum leaf extract exhibited the highest inhibition percentage at 66.10±3.22%. These findings indicate that all three plant extracts contain compounds capable of inhibiting PLpro activity
Population Growth Model and Mortality of Pakistan Lobster (Panulirus polyphagus) in Estuary Waters of Tarakan City
Get PDFLobster Panulirus polyphagus has a fairly high economic value and is found in the estuary waters of Tarakan City. This research aims to study the growth and mortality model of Lobster P. polyphagus originating in the estuary waters of Tarakan City. The research method was carried out using a quantitative descriptive method. Sampling was conducted 14 times from December 2021-May 2022 using gill nets. The results showed that the male sex ratio was more than the female. Allometric growth of males and females is negative allometric with a thin body shape. The structure of the size obtained was mostly in males ranging from 18.3-20.6 cm, and females around 20.5-22.2 cm. The maximum length growth of P. polyphagus based on von Bertalanffy\u27s growth model was 31.519 cm in males and 31.374 cm in females. The total mortality (Z) of P. polyphagus for males and females was 1.104 and 1.119; catch mortality (F) of 0.106 and 0.253; natural mortality (M) of 0.998 and 0.866; exploitation rate (E) of 0.096 and 0.226, respectively. The high natural mortality causes the extinction of the Lobster species, so good management is needed so that it is sustainable