APPLICATION OF IMAGE-BASED HIGH CONTENT ANALYSIS FOR THE SCREENING OF BIOACTIVE NATURAL PRODUCTS

Each bioactive compound induces phenotypic changes in target cells that can be made visible by labelling selected molecules of the cells with fluorescent dyes and/or directly observed under the high-throughput microscope. A comparison of the cellular phenotype induced by a compound of interest with known cellular targets allows predicting its mode of action. Over the past 15 years, high-throughput microscopy has been one of the fastest growing fields in cell biology. When combined with automated multiparametric image and data analysis, it is referred to as high-content screening (HCS). Whilst HCS has been successfully applied to the bioactivity characterization of natural products, recent studies used automated microscopy and software to increase speed and to reduce subjective interpretation. In 2017, Institute of Natural Products Chemistry (INPC-VAST) has been equipped with a HCS platform (Olympus Scan^R) that designed for fully automated image acquisition and analysis of biological samples to visually inspect the cellular morphology induced by hit compounds as well as to discriminate from false positives. Accordingly, this short review covers the concepts of HCS and its application in screening of biologically active natural products whose molecular targets could be identified through such approaches

USING EXPERIMENTAL PLANNING TO OPTIMIZE THE HYDROLYSIS OF SUGAR CANE BAGASSE INTO FERMENTABLE SUGARS FOR BIOETHANOL PRODUCTION BY FUNGAL ENZYME MIXTURE

In this study, the unpretreated sugar cane bagasse was milled to a particle size of 0.5 – 1 mm and be used as material for bioconversion into fermentable sugars by using an enzyme cocktail acted synergistically. Experimental planning was used to optimize the enzyme conversion through assessment and analysis of individual parameter. As the result, the optimal condition for enzymatic conversion of sugar cane bagasse into reducing sugar product are at pH = 5, 400C, and 48 h incubation in rate of enzyme (Cell/Xyl, AltFAE, XpoAE) and substrate (bagasse meal) is 3.1. From the above application, the mathematical model is found to describe equation of the bioconversion of bagasse into reducing sugars: = 206.946 + 29.954x1 + 5.501x2 + 7.323x3 + 2.288x2x3 – 7.011; and using flexible algorithm of nonlinear planning to identify optimal conditions of enzyme mixture of conversion into reducing sugars that the reaction reached max = 251.86 mg per gram bagasse with x1 = 1.215, x2 = 1.215, x3 = 1.215 or Cell/Xyl = 1 ml (100U), AltFAE = 0.5 ml (7.56U), XpoAE = 0.4 ml (10.8U) on the test range. Experimental verification has the same result in constant conditons and reached total reducing sugars of 260.2 mg per gram substrate.In this study, the unpretreated sugar cane bagasse was milled to a particle size of 0.5 – 1 mm and be used as material for bioconversion into fermentable sugars by using an enzyme cocktail acted synergistically. Experimental planning was used to optimize the enzyme conversion through assessment and analysis of individual parameter. As the result, the optimal condition for enzymatic conversion of sugar cane bagasse into reducing sugar product are at               pH = 5, 40 oC, and 48 h incubation in rate of enzyme (Cell/Xyl, AltFAE, XpoAE) and substrate (bagasse meal) is 3.1. From the above application, the mathematical model is found to describe equation of the bioconversion of bagasse into reducing sugars: = 206.946 + 29.954x1 + 5.501x2 + 7.323x3 + 2.288x2x3 – 7.011; and using flexible algorithm of nonlinear planning to identify optimal conditions of enzyme mixture of conversion into reducing sugars that the reaction reached max = 251.86 mg per gram bagasse with x1 = 1.215, x2 = 1.215, x3 = 1.215 or Cell/Xyl = 1 ml (100U), AltFAE = 0.5 ml (7.56U), XpoAE = 0.4 ml (10.8U) on the test range. Experimental verification has the same result in constant conditons and reached total reducing sugars of 260.2 mg per gram substrate

Isolation and identification of indole acetic acid producing bacteria from the coasts of Ben Tre and Tra Vinh Provinces

Beneficial plant-growth-promoting bacteria (PGPB) have been reasonably applied to rescue crucial issue for agriculture by salinity soil. Observed most of PGPB was found in endophyte, rhizosphere and soil. Indole acetic acid (IAA)-producing bacteria could naturally stimulate and facilitate plant growth. The knowledge of IAA production and content of bacteria resident in the marine environment has been typically insufficient and limited to date. In recent years, unwarrantable intrusions of sea water have been enlarged in the Mekong River Delta of Vietnam, threatening productive rice fields, local fruits, and cash crops. Therefore, finding PGPB in the coastal regions in the Mekong River Delta as a creative resource for sustainable agriculture is necessary and is a prompt challenge. In this study, IAA-producing bacteria from coastal regions of Ben Tre and Tra Vinh Provinces were isolated and adequately identified. Out of 202 bacterial isolates, 10 isolates showed the possible ability to produce IAA from L-tryptophan. These 10 isolates were objectively evaluated the capacity to produce IAA under 5% (w/v) NaCl in King B and marine broths. The results revealed that IAA production decreased in 5% NaCl, even though bacterial growth increased. These 10 IAA-producing bacteria were classified at the species level, Marinobacter hydrocarbonoclasticus, M. pelagius, M. daepoensis, and Mameliella phaeodactyli by 16S rRNA gene analysis. The most IAA producer in King’s B broth, the isolate C7, was investigated in more detail. The isolate C7 produced the maximum IAA amount (192.2 ± 1.14 µg/ml) under the presence of 20 g/l yeast extract, 2 g/l of L-tryptophan and 1% NaCl. The isolate C7 was able to grow at 1–17% (w/v) NaCl (optimum, 4%), but not in the absence of NaCl, indicating it is a moderate halophilic bacteria. This study highlighted the considerable ability to produce IAA of marine bacteria, which could be thoughtfully considered to use naturally as biofertilizers to promote plant growth in saline intrusion lands.

A modern purification by accelerated solvent extraction and centrifugal partition chromatography and biological evaluation of capsaicin from Capsicum chinense

A special alkaloid compound known as capsaicin, which can only be found in the fruit of the Capsicum plant, was isolated and tested for its anti-inflammatory activity. The purpose of this work is to establish a simple and quick approach for capsaicin purification utilizing centrifugal partition chromatography (CPC) as well as an effective method - accelerated solvent extraction (ASE), for extracting capsaicin from Capsicum chinense. After purification, capsaicin was validated by HPLC-DAD at 281 nm to be > 90% purity. The in vivo anti-inflammatory activity of the isolated capsaicin was also investigated, and the IC50 value of the capsaicin was determined to be 57.61 µg/mL. The current work emphasizes how an ASE and CPC system may combine to extract high-purity capsaicin from Capsicum chinense, which have the anti-inflammatory activity, as we evaluated in the experiment

The Effectiveness of Narrow Band Uvb (Nb-Uvb) In the Treatment of Pityriasis Lichenoides Chronica (PLC) In Vietnam

AIM: This prospective clinical study presents the experiences with NB-UVB monotherapy in the treatment of PLC on Vietnamese patients. METHODS: We enrolled at National Hospital of Dermatology and Venereology (NHDV), Vietnam, 29 PLC patients with generalised disease involving at least 60% of the total body surface (based on Nine’s Rule) and/or failed to respond to other modalities of treatment. Patients were treated with NB-UVB followed the guideline of the psoriatic treatment of AAD-2010, three times weekly. RESULTS: A complete response (CR) was seen in 24 out of 29 PLC patients (82.8%) with a mean cumulative dose of 9760.5 mJ/cm2 after a mean treatment period of 4.6 weeks (13.8 ± 7.4 exposures). Mild side effects were observed: 69% erythema minimum, 55.2% irritation related to dry skin. No severe side effects were seen during the study. No relapses occurred in 24 CR patients within a mean period of 3 months after the last treatment. CONCLUSION: NB-UVB therapy is an effective and safe option for the treatment and management of PLC

The Decline of PUVA Therapy in Vietnam: Effective Treatment of Narrow Band UVB in Vietnamese Vitiligo Patients

AIM: To examine the efficacy and safety of Narrowband ultraviolet B (NB-UVB) in Vietnamese vitiligo patients. METHODS: We recruited thirty-one patients (14 males, 17 females), aged from 7 to 67 years, with both segmental vitiligo (SV) and non-segmental vitiligo (NSV), treated three times weekly with NB-UVB. The starting dose for adults from 15 years old and children less than 15 years old was 200 mJ/cm2 and 150 mJ/cm2, respectively, with 50 mJ/cm2 and 20 mJ/cm2 dose increments at each subsequent visit, respectively, until mild erythema lasting less than 24 hrs reported by patient, given for a period of 6 months. Response to therapy was assessed based on VASI score changes. RESULTS: Based upon our results, 38.7% (12/31) of patients achieved a very good response of more than 50% VASI changes, 41.9% (13/31) obtained a good response (VASI changed from 25 to 50%). Total good and very good response to therapy significantly increased with prolonged treatment, increasing from 19.4% to 64.5% and 80.6% after 2, 4 and 6 months, respectively. Localised NSV patients obtained good and very good response significantly more frequently than generalised NSV (55.6% versus 18.2%). Adverse effects were minimal, of which one case developed herpes simplex, and 4 cases reported mild photo burn reaction which completely disappeared after adjusting the dose. CONCLUSION: NB-UVB therapy is an effective and safe tool in the management of Vietnamese vitiligo patients

Developing Application Based Upon An Ontology-Based Modelling of Vietnamese Traditional Dances

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