Protective Signatures of Roselle (Hibiscus sabdariffa L.) Calyx Fractions against Staphylococcus aureus in Drosophila Infection Model

The rise of antibiotic-resistant Staphylococcus aureus-related clinical cases is an alarming chronicle for global communities. This research was conducted to examine the antistaphylococcal effect of roselle (Hibiscus sabdariffa L.) calyx fractions in the Drosophila model. In the infection experiment, wild-type and immunodeficient Drosophila were pricked with S. aureus and subsequently subjected to fly survivorship and colony-forming assays, in the presence or absence of roselle calyx fractions. The Involvement of immune stimulation in the host antibacterial protection was assessed in vitro using cell-based luciferase reporter assay and in vivo using RT-qPCR analysis on adult flies. A declining rate of fly survivorship and augmentation of bacterial growth were observable in S. aureus-infected wild-type flies but subject to improvement in the presence of roselle calyx fractions. Cell-based analysis revealed the absence of host immune stimulation via Drosophila Toll pathway and roselle calyx fractions-treated immune-deficient flies lacking for components in the Toll pathway were protected from infection-induced early death phenotype and harbored reduced number of S. aureus colonies. Overall, our data confirmed the in vivo anti-staphylococcal activity of roselle calyx fractions in Drosophila infection model and such protective signature was devoid of host immune stimulation

Determination of Genetic Mutation Profile of drpr Gene in Drosophila melanogaster using High-Resolution Melting Analysis

Genotype determination of experimental animals is generally conducted using sequencing methods that require expensive cost as well as experience and special equipment. This study aimed to determine the presence of drpr gene mutation in Drosophila melanogaster using coupled real time PCR-High Resolution Melting (real time PCR-HRM) as an alternative method. Two types of fly samples, w1118 and drprΔ5 were used as wildtype control and mutant genotype, respectively. The DNA from twenty of each w1118 and mutant drprΔ5 flies were isolated and amplified using real time PCR. The generated amplicons were then further processed by HRM method at the temperature of 60-95°C. This study demonstrated that the real time PCR-HRM method could distinguish wildtype control w1118 and mutant drprΔ5 based on the HRM data with the confidence level was more than 90%. Therefore, this study provides an evidence that real time PCR-HRM method might be beneficial to screen the mutant genotype from its wildtype counterpart based on differences in the melting temperatures due to changes at nucleotide base leve

Determination of Genetic Mutation Profile of drpr Gene in Drosophila melanogaster using High-Resolution Melting Analysis

Genotype determination of experimental animals is generally conducted using sequencing methods that require expensive cost as well as experience and special equipment. This study aimed to determine the presence of drpr gene mutation in Drosophila melanogaster using coupled real time PCR-High Resolution Melting (real time PCR-HRM) as an alternative method. Two types of fly samples, w1118 and drprΔ5 were used as wildtype control and mutant genotype, respectively. The DNA from twenty of each w1118 and mutant drprΔ5 flies were isolated and amplified using real time PCR. The generated amplicons were then further processed by HRM method at the temperature of 60-95°C. This study demonstrated that the real time PCR-HRM method could distinguish wildtype control w1118 and mutant drprΔ5 based on the HRM data with the confidence level was more than 90%. Therefore, this study provides an evidence that real time PCR-HRM method might be beneficial to screen the mutant genotype from its wildtype counterpart based on differences in the melting temperatures due to changes at nucleotide base leve

Efek Sinergitas Ekstrak Kulit Jeruk (Citrus sinensis L) Pada Patch Bioselulosa Dalam Meningkatkan Penyembuhan Luka Bakar

Burns is one of the incidents that can lead to death (mortality). One of the natural products that have  potential to serve as an alternative treatment of burns is orange peel. Orange peel has a chemical composition such as ascorbic acid, vitamin E, vitamin A, and polyphenols as antioxidants that inhibit free radicals responsible in the pathogenesis of both acute and chronic inflammatory. In this study, formulation was made in the form of biocellulose which is the primary metabolism product of bacteria.  The purpose of this research was to obtain the concentration of the extract of orange peel on bioselullose that have the effect of decreasing the burn wound in rats. Orange peel was extracted then fortified into biocellulose with a concentration of 3%, 6%, and 9%. After that, the wound healing was tested on animals in the form of decreasing the wound diameter.  The results showed that the concentration of extract of orange peel 3% on the fortification of biocellulose showed the good percentage of burn wound decreasing i.e. 45.52% with diameter average of 18.35 mm. This result indicates the concentration of extract of orange peel 3% is better than others

Ex Vivo and In Vivo Retention Time Evaluation of Fucoidan Isolated from Macrocystis pyrifera Through a Thermosensitive Gel System in The Vaginal Route

This study evaluated Fucoidan from Macrocystis pyrifera as a potential treatment for cervical cancer. The research aimed to examine Fucoidan’s in vivo retention capacities in poloxamer-based in situ gels for vaginal drug delivery systems. Five different thermosensitive gel formulations were developed, each with varying concentrations of Pluronic F127 and F68 polymers. The incorporation of HPMC affected the gelation temperature, viscosity, and bioadhesive strength. The accepted formula, F3, had a bioadhesive value of 5415.93 ± 98.74 dyne/cm2 and could form a gel at physiological temperature. Ex vivo animal models showed that Fucoidan components retained well on vaginal tissue. Only F1, F2, and F3 achieved the media after 8 hours of examination. In vivo evaluation showed F3 had the highest drug concentration retained in the vaginal mucosa of female rats after 8 hours (24,115 ± 4,842 g), slowly removed after 24 hours (13,014 ± 5,596 g). In conclusion, increases in the hydrophilic content of formulations led to the retained hydrogel formula, which increased drug release and lowered intravaginal elimination