EVALUATION OF ANTI-GLYCATION EFFECT AND SAFETY OF SERUM ANTI-AGING FORMULATION CONTAINING GOLD NANOPARTICLES (AUNP) USING SIDAGURI EXTRACT (SIDA RHOMBIFOLIA)

Objective: The purpose of this study is to determine the anti-aging properties and safety of serum containing gold nanoparticles (AuNP) using Sidaguri extract (Sida rhombifolia) through their anti-glycation effect. Methods: The anti-aging effect of serum was performed in vitro by measuring advance glycation end products (AGEs) formed during incubation using a Microplate reader, and safety of serum was performed using hen’s egg chorioallantoic membrane (HET-CAM) method using White Leghorn egg. Results: The study showed that serum formulation had an anti-glycation effect with inhibition percentages are 68.20±6.86% and 74.83%±19.91% for a serum containing 10% and 20% gold nanoparticles and little to no irritation potency for both serum formulations with RI value 0.0 and 0.0, respectively. Conclusion: Due to both their anti-glycation effect and irritation behavior, serum formulation containing gold nanoparticle synthesized using Sidaguri extracts could be utilized as anti-aging cosmetics in the future.

Detecting Redox Potentials Using Porous Boron Nitride/ATP-DNA Aptamer/Methylene Blue Biosensor to Monitor Microbial Activities

Microbial activity has gained attention because of its impact on the environment and the quality of people’s lives. Most of today’s methods, which include genome sequencing and electrochemistry, are costly and difficult to manage. Our group proposed a method using the redox potential change to detect microbial activity, which is rooted in the concept that metabolic activity can change the redox potential of a microbial community. The redox potential change was captured by a biosensor consisting of porous boron nitride, ATP-DNA aptamer, and methylene blue as the fluorophore. This assembly can switch on or off when there is a redox potential change, and this change leads to a fluorescence change that can be examined using a multipurpose microplate reader. The results show that this biosensor can detect microbial community changes when its composition is changed or toxic metals are ingested.

Synthesis of 5(S)-Methyl-L-Proline containing Peptidomimetic Compounds and their In-Vitro Evaluation for Dipeptidyl Peptidase-4 Inhibition

Background: Type 2 diabetes mellitus (T2DM), which is the epidemic of the 21st century, has affected millions of people worldwide. Traditional methods available for the treatment are associated with various side effects. Among the newer therapies, DPP-4 (Dipeptidyl peptidase-4) inhibition has been a promising therapy for the past decade with the scope of further development, especially in peptidomimetics. Objective: 5(S)-methyl-L-proline containing peptidomimetic compounds were designed in the previous work. The designed compounds were synthesized and characterized by spectral methods, such as mass spectrometry, 1H NMR, and 13C NMR (Nuclear magnetic resonance) spectroscopy. The purity of the final compounds was determined by high-performance liquid chromatography (HPLC). The synthesized compounds were in vitro evaluated for their DPP-4 inhibitory activity. Method: Compounds were peptide in nature and were synthesized using the conventional synthesis approach, where peptide synthesis was done using an acid-amine coupling reagent. They were evaluated through fluorimetric enzyme-based assay using a DPP-4 inhibitor screening kit. Moreover, the CLARIOstar microplate reader instrument was used to measure fluorescence. Results: 5(S)-methyl-L-proline containing 13 compounds were synthesized. All of them were characterized for structural integrity using spectral methods. They had HPLC purity of more than 95% and were evaluated for DPP-4 inhibition. Compounds 001, 007, 010, 011, 014, and 017 were found to have good inhibition than others. These compounds were further evaluated at different concentrations to develop a linear correlation coefficient (R2). Conclusion: Six compounds were found to have good DPP-4 inhibition, hence it further opens the possibility of developing DPP-4 inhibitor-containing 5(S)-methyl-L-proline.

Uji Aktivitas Ekstrak Daun Dan Akar Singawalang (Petiveria alliaceae) Terhadap Penghambatan Tirosinase

Tyrosinase is an enzyme that plays a role in the formation of skin pigments from a person because it is involved in the process of melanogenesis. Tyrosinase plays a very important role in the skin depigmentation process, there are several attempts to inhibit the skin depigmentation process, one of which is by inhibiting tyrosinase. Research on the leaves and roots of singawalang (Petiveria alliacea) was conducted to determine the potential as a tyrosinase inhibitor. Leaves and root extracts of singawalang were macerated with ethanol, then tested for identification of secondary metabolites. Singawalang leaves extract contains alkaloids, tannins and terpenoids while singawalang root extract contains alkaloids, flavonoids, tannins and terpenoids. Tyrosinase inhibitory activity used the microplate reader ELISA technique at a wavelength of 492 nm. Tests were carried out on kojic acid as a comparison and L-DOPA as a substrate. The results of the tyrosinase inhibition activity test for the extracts of singawalang leaves, singawalang roots and kojic acid, IC50 were 9.817 mg / mL, 4.987 mg / mL and 0.092 mg / mL, respectively.

Comparative Study Highlights the Potential of Spectral Deconvolution for Fucoxanthin Screening in Live Phaeodactylum tricornutum Cultures

Microalgal biotechnology shows considerable promise as a sustainable contributor to a broad range of industrial avenues. The field is however limited by processing methods that have commonly hindered the progress of high throughput screening, and consequently development of improved microalgal strains. We tested various microplate reader and flow cytometer methods for monitoring the commercially relevant pigment fucoxanthin in the marine diatom Phaeodactylum tricornutum. Based on accuracy and flexibility, we chose one described previously to adapt to live culture samples using a microplate reader and achieved a high correlation to HPLC (R2 = 0.849), effectively removing the need for solvent extraction. This was achieved by using new absorbance spectra inputs, reducing the detectable pigment library and changing pathlength values for the spectral deconvolution method in microplate reader format. Adaptation to 384-well microplates and removal of the need to equalize cultures by density further increased the screening rate. This work is of primary interest to projects requiring detection of biological pigments, and could theoretically be extended to other organisms and pigments of interest, improving the viability of microalgae biotechnology as a contributor to sustainable industry.

Forms of infectious complications after use of metal foreign bodies of the auricle and determination of the ability of detected microorganisms to biofilm formation

Ear or nose are quite vulnerable to foreign bodies. Foreign bodies cause various side effects in the body. A special category consists of foreign bodies that are introduced for aesthetic purposes, in particular piercing products. Decoration implanted in the tissues of the ear or nose is a foreign body and is a field of increased contamination and reproduction of pathogenic and opportunistic microorganisms. The aim of the study was to determine the infectious forms of complications that occurred after prolonged use of metal foreign bodies and to study the ability of biofilms by microorganisms isolated from the pathological contents of the foreign body of the auricle. 93 patients who applied to the ENT department of Kharkiv Regional Hospital for complications after implantation of metal foreign bodies and 10 people, control group, which had no foreign bodies, and were randomly selected from healthy individuals, were examined, to determine the qualitative composition of the microbiocenosis of the auricle skin. The material for the study was pathological discharge from a foreign body of the auricle. The microbiological study was performed using MICRO-LA-TEST identification kits. Studies of the formation of biofilms were studied by determining the ability of bacterial strains to adhere to the surface of polystyrene. The obtained cultures were washed with suspension media individual for each family of bacteria. The optical density of bacterial suspensions was measured using a microplate reader "MultiskanEX" (type 355). Statistical analysis of the obtained data was performed using MSExcel, Statisica 10 software. As a result of the conducted researches the dependence between the development of infectious complications of the auricle in the presence of metal foreign bodies and the microbiocenosis of the pathological focus was revealed. The study allowed to establish the structure of the microbial landscape of the skin of the auricle in the area of the metal foreign body, to determine the dominant forms of complications of infectious origin, after prolonged use of metal foreign bodies and to study the ability to form biofilms by microorganisms from different metals. The ability of various types of microorganisms to form biofilms when using products from different types of metals has been studied. It is established that the use of metal foreign bodies made of silver and gold reduces the risk of purulent-inflammatory process. It is proved that the optical density of biofilms of most microorganisms isolated from the pathological contents of the area of foreign bodies made of silver and gold is significantly lower than when using steel and titanium products.

FPCount protocol - Full protocol v1

FPCount is a complete protocol for fluorescent protein calibration, consisting of: 1. FP expression/purification using Thermo's HisPur Cobalt Resin. 2. FP concentration determination in a microplate reader. 3. FP fluorescence quantification in a microplate reader. Results can be analysed with the corresponding R package, FPCountR. --- Summary 1. Expression 2. Harvesting/Washing 3. Lysis 4. Fractionation 5. Gel1: Verification of Expression/Fractions 6. Purification 7. Gel2: Verification of Purification 8. Protein concentration and buffer exchange 9. Quantification of FP concentration (part1) 10. Quantification of FP fluorescence 11. Quantification of FP concentration (part2) 12. Protein Storage 13. Calibration of Plate Reader