HAYATI Journal of Biosciences
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Development of Phenol Red Colorimetric RT-LAMP Assay in High-Buffered SARS-CoV-2 Sample
Get PDFColorimetric RT-LAMP Assay is a diagnostic method that has attracted much attention because of its rapidity, simplicity, and accuracy compared to other disease diagnosis methods. Despite having many advantages, the RT-LAMP Colorimetric Assay has disadvantages, especially for kits that use phenol red as an indicator. The disadvantage derives from the input RNA/DNA samples containing high buffer levels, which causes no color change and false-negative results. This study aimed to develop and optimize the colorimetric RT-LAMP method on high-buffered SARS-CoV-2 RNA samples. We found that a temperature of 69°C for 50 minutes with the addition of post-treatment in the form of heating at 80°C for 10 minutes is an optimal condition for high-buffered SARS-CoV-2 samples. The condition proved effective in changing the result\u27s color from red (negative) to yellow (positive). We also classified the analysis results based on the correlation between the Cycle threshold (Ct) value of SARS-CoV-2 viruses and the Optical Density (OD) value, which was quantified using a spectrophotometer at 415 nm (with a correlation value of -0.9084), where yellow color indicated Ct below 20, amber color indicated Ct between 20 and 30, orange color indicated Ct between 30 and 35, and red indicated Ct more than 35 (negative). In conclusion, this study successfully detects the SARS-CoV-2 virus in high-buffered samples using Phenol Red Colorimetric RT-LAMP Assay, with a sensitivity of 85% for Ct Cutoff 40
Chemical Composition, Antioxidant, Antibacterial, Antibiofilm, and Cytotoxic Activities of Robusta Coffee Extract (Coffea canephora)
Get PDFCoffee extracts are a rich source of potential compounds with numerous biological activities. This study aimed to investigate the potential of chemical compounds derived from robusta coffee extract (Coffea canephora) and its dominant compound, caffeine, to scavenge free radicals, inhibit bacterial growth and biofilm formation, and cytotoxic properties in human breast adenocarcinoma cell line. Chemical constituents of coffee extract were analysed quantitively for total phenolic, flavonoid, and alkaloid contents, along with Liquid Chromatography Quadrupole-Mass Spectrometry (LC/MS–MS) analysis. Antioxidant activity was measured by 2,2-Diphenyl-1-picrylhydrazyl (DPPH) and 2,2\u27-azino-bis; 3-ethylbenzothiazoline-6-sulfonic acid (ABTS) methods. Antibacterial pair with antibiofilm properties against Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, and Staphylococcus aureus were also determined, as well as cytotoxic activity on the MCF7 cell line model. The LC/MS–MS analysis of coffee extracts revealed high levels of caffeine; thus, the caffeine standard is used in all subsequent assays. Notably, robusta coffee extract showed remarkable antioxidant activity and selective inhibition of the growth against gram-positive and negative bacteria, along with the best inhibition of P. aeruginosa biofilm. However, compared with the caffeine standard, robusta coffee extract had lower cytotoxic activity and different bacterial targets in antibacterial and antibiofilm properties. Our results indicate that Robusta coffee extract is potentially a functional food due to its high alkaloid, phenolic, and flavonoid content and antioxidant activity, besides being used for natural compounds against bacterial infections
Antifungal Substances Produced by B. subtilis Strain W3.15 Inhibit the Fusarium oxysporum and Trigger Cellular Damage
Get PDFSoybean Fusarium wilt and root rot disease caused by a necrotrophic ascomycete pathogen, F. oxysporum, triggered severe damage to the plant tissues and organs and impacted heavy losses. Biocontrol agents, Bacillus subtilis, were commonly used to produce a broad spectrum of antifungal substances and were gradually used in biocontrol studies for plant disease management. Investigation and determination of the inhibiting mechanism of antifungal substance produced by B. subtilis on F. oxysporum should be done to protect the soybean plant. This study revealed that basal nutrient broth (NB) gives the best antifungal activity. The stationary phase of the bacterial growth curve was obtained on two days of cultivation and showed the maximum antifungal activity against F. oxysporum. Ethyl acetate (EA) extraction of bacterial supernatant generated crude EA extract, which showed half inhibition (IC50) at 306.42 µg/ml obtained from the dose-response regression curve. Post-treatment mycelia of F. oxysporum with bacterial extract were demonstrated as hyphal deformation followed by malondialdehyde (MDA) accumulation. Furthermore, cellular leakage on fungal cells that may be triggered by antifungal compounds from strain W3.15 occurred. Last, the related antifungal compounds were predicted to be epicatechin and benzophenone from the LC-MS/MS analysis of crude EA extract. Accordingly, the biocontrol agent B. subtilis strain W3.15 promises a strong potency for biofungicide development
Formulation of Anti-Oral Mouthwash Nanoemulsion Biofilm Based on Propolis Extract Heterotrigona itama, Tetragonula sapiens, and Tetragonula clypearis
Get PDFThe use of mouthwash is one of the actions against biofilms that are often used. However, commercial mouthwashes have a fairly high alcohol content, which is around 26.9% of the total volume, which is considered to have a prolonged impact where high alcohol content in direct contact with the oral mucosa can cause lesions or abnormalities, resulting in a shift in the medical paradigm towards eco-friendly widely considered as a solution. Propolis with antibacterial ability was formulated using the nanoemulsion steps, which were initiated by separating pure propolis through drying, and then there were variations in the formulation of 3 types of bee propolis: Heterotrigona itama, Tetragonula sapiens, and Tetragonula clypearis along with the addition of Tween 80, propylene glycol, glycerin and then the effect on microbial growth of S. mutans was compared with antimicrobial agents in Brazilian propolis with the identification and comparison of the antibacterial activity stability of the organoleptic formula. Where the active ingredient content of propolis is the highest in H. itama propolis with a total flavonoid content of 38.94 mgQE/L sample and T. clypearis propolis has the lowest total flavonoid content of 14.23 mgQE/L sample with its function as an anti-oral biofilm agent by inhibiting the glucosyltransferase was proven with a minimum percentage of 49% inhibition of S. mutans and degradation of 18% with the use of a combined surfactant proved to be able to increase the stability of the preparation shown at 2:1 (v/v)
Topical Application of Paraboea leuserensis on Excision Wound with Angiogenesis and Vascular Endothelial Growth Factor Analysis
Get PDFGagatan harimau or Paraboea leuserensis B.L. Burtt is an endemic plant that grows in limestone areas, natively found in Leuser Mountain Area, North Sumatra Province, Indonesia. This study determines the wound healing potential of P. leuserensis gel-based ethanolic extract (PGEE) on full-thickness excision wounds based on the angiogenesis and VEGF expression. Here, we reported 67 compounds detected by LC-MS/MS from the ethanolic extract. The antibacterial activity regarding pathogenic wound infection was also investigated. The experimental study used 30 Rattus novergicus, randomly divided into five treatment groups: base gel as a negative control, Bioplacenton as a positive control, PGEE 2.5%, 5%, and 10%. Experimental data showed that PGEE indicated a significant effect on bacterial inhibition (p = 0.02), where the greatest inhibitory was found against S. epidermidis. Topical application of PGEE showed significant differences in the number of blood vessels of PGEE 2.5% and 10% (p = 0.007) and expression of VEGF (p<0.05). The correlation between the number of blood vessels and VEGF was also described with the number of macrophages, fibroblasts, collagen, and wound diameter. Overall, this study demonstrates the pharmacological potential of PGEE for wound healing applications regarding angiogenesis and VEGF expression
New Proposed Dose of Irradiation to Control Fruit Flies (Diptera:Tephritidae) and its Influence to Super Red Pittaya (Hylocereus costaricensis) Quality
Get PDFAs one of the important pests, fruit flies derived from Tephritidae family have caused the worst consequences in losing the economic value of fresh horticultural commodity including on the dragon fruits. As a prospective quarantine treatment, gamma irradiation has been thrived to eliminate totally the appearance of those adults’ flies with less impact on the fruit quality degradation. In this study, a series of irradiation dosages were examined to both the third instar larvae (subsequently from Bactrocera papayae, B. cucurbitae and B. carambolae) infesting fruits and the non-infested ones afterwards at 0, 50, 100, 150, 200, 250, 300, 350 and 400 Gy. For a number of the infested fruits, a series of observations were persisted in at room temperature (27±1°C) until two weeks of the post-treatment period. Whereas the non-infested fruits were stored at 11±1°C for 17 days of the storage. Results showed that B. papayae required the highest recommended dosages (the effective dose = 248.4 Gy; the minimum dose = 225.1 Gy; the maximum dose = 279.6 Gy) for achieving LD0.99 rather than the other tested spesies. There were also no significant differences among each of the non-infested fruits in terms of altering of the observed physical attributes. This result has become a distinct indication that a higher dose value than the generic one is mostly recommended both for phytosanitary irradiation and stabilization of fruit quality
Cardioprotective Activity of Costus Root Ethanol Extract in Experimentally-Induced Hypothyroidism in Female Albino Rats
Get PDFHypothyroidism is a popular endocrine disorder caused by a thyroid hormone deficit. Changes in the state of the thyroid gland significantly affect many organs, including the heart. This study examined the cardioprotective effects of an artificially induced hypothyroidism in female rats using an ethanol extract from the root plant Saussurea lappa (Costus). For the current study, 25 adult female albino rats were employed. They were split into five equal groups, including control, hypothyroid group, post-treatment group with costus extract, co-administered hypothyroid group with costus extract, and post-treatment group with levothyroxine. Thyroid hormones (T3, T4, and TSH), lipid profile, and oxidative stress indicators (catalase, superoxide dismutase, reduced glutathione, and lipid peroxidation levels) were also measured in the serum. The heart muscle was the subject of a histological research. The findings suggested that Costus root ethanol extract improved hypothyroidism in female rats, as proven by the reversal of many biochemical abnormalities and improvements in the heart\u27s histology. Our study indicates that the root of Costus exhibited cardioprotection efficiency against hypothyroidism-induced serious effects on the heart, The antioxidant and radical-scavenging properties of its components may be responsible for this promising effect
Bird Sounds Psychological Restorative Effect on The Visitors of Bogor Botanical Garden
Get PDFStress can be defined as psychological and physical tension caused by physical, emotional, social, economic, and occupational, events, or experiences that are difficult to manage or endure. Bird sounds can be a potential solution to reduce stress levels. This study aimed to analyze the effect of bird sounds as a stress restorative on Bogor Botanical Garden visitors. A survey of 100 visitors of Bogor Botanical Garden was conducted based on the Perceived Stress Scale and Perceived Restorativeness Soundscape Scale. Data on respondent characteristics were analyzed using descriptive analysis, the relationship between stress levels and perceived stress restorative effect of bird sound was analyzed using Pearson correlation analysis. The analysis showed that 78% of respondents had moderate to high perceived stress levels and 98% of respondents rated the degree of restoration from bird sounds to be moderate to high. The correlation analysis revealed a negative relationship between the stress level and bird sounds\u27 perceived restorative effect. The older the respondents and the more frequently they visited Bogor Botanical Garden, the higher the perceived stress restorative effect from bird sound
Antioxidant, Cytotoxic Activity and Protein Target Inhibition of Ethyl Acetate Fraction Melinjo Seed (Gnetum gnemon L.) by In Vitro and In Silico Studies on HeLa Cervical Cancer Cells
Get PDFCervical cancer is one of the most common cancer suffered in women. Chemotherapy usage often causes physical and psychological side effects in patients. This study aims to determine the antioxidant and cytotoxic effects of the ethyl acetate fraction of melinjo seeds (Gnetum gnemon L.) on HeLa cervical cancer cells through in vitro and in silico assays. Melinjo seed was extracted by maceration using ethanol 70% and fractionated with ethyl acetate to obtain the Ethyl Acetate Fraction of Melinjo Seed (EAFMS). The identification of the active compounds group was done using Thin Layer Chromatography (TLC) method. In vitro studies were conducted on antioxidant tests using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) method and cytotoxic activity test using 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide (MTT) Assay. In silico test for molecular docking analyzed by Autodock Vina method. The TLC analysis of EAFMS showed the presence of the stilbenoid compounds group. The antioxidant activity of EAFMS is weak, with an IC50 value of 175.8 g/ml. Cytotoxic activity of EAFMS is categorized as toxic to HeLa cancer cells with an IC50 value of 21.69 g/ml, while EAFMS has a synergistic effect combined with doxorubicin as a standard drug with a combination index (CI) value of 0.24-0.80. A molecular docking test of gnetin C with VHR receptor found a strong and stable bond with a docking score of -8.3 kcal/mol. Thus, EAFMS has the potential to be used as a chemopreventive agent for cervical cancer and can be combined with doxorubicin
Investigation of Morinda citrifolia Activity Related Collagen Type II Synthesis Through Gene Expression
Get PDFTo extend plant benefits, three different concentrations of five Morinda citrifolia part was investigated on the collagen type II which is the primary collagen in human cartilage through the expression of the genes, COL2A1, COL-II and COLL2 regions in normal human dermal fibroblasts by qRT-PCR method. The results showed that 1) fibroblasts cultured in the presence of M. citrifolia extracts produced many times more collagen type II gene expression than control cells depending on the plant parts and concentrations, 2) the expression levels of the collagen type II gene stimulated by all fruit parts yields the higher percentages than leaves. Next, Morus alba leaf extract at a concentration expected to be a precursor protein source for collagen synthesis, working together with M. citrifolia stimulation, was added to the selected concentration indicating high expression in the M. citrifolia fruit experiments. The result showed various gene expression levels depended on the kind of gene and fruit part. Therefore, M. citrifolia fruits can benefit the creation of collagen type II with or without M. alba. The M. citrifolia fruit can be further benefited in product production for both the elderly and young for maintaining the typical structure and function of the skin, tendon, and bone