We aimed to evaluate the effects of solid-state fermentation (SSF) on red rice bran, specifically for a new rice variety (RD69,Tubtim Chumphae) in Thailand. Fermentation of red rice bran with Aspergillus oryzae was continued for 6 days at 30 °C, and LC MS/MS was used to determine the free amino acid composition, antioxidant activity of 2,2-diphenyl-1-picrylhydrazyl, ferric reducing antioxidant power, presence of phenolic and flavonoid compounds, and cytotoxicity in HeLa cells. The results showed that SSF significantly increased (p < 0.05) the total amino acid content by 59.00% after 5 days of fermentation. Fermented red rice bran (FRRB) contained a significant increase (p < 0.05) in antioxidant activity and compounds, which reached their maximum levels by fermentation for 5 days. FRRB also exhibited increased cytotoxicity in HeLa cells compared with unfermented rice bran. The increases of cytotoxicity and antioxidant activities were due to enzymatic hydrolysis and fermentation, in correspondence with the increase in phenolic and flavonoid compounds, suggesting that enzymatic hydrolysis improves the nutritional and phytochemical properties. Notably, the duration of fermentation is a key to optimizing the biological activity of the fermented product.
Evaluation of the amino acid composition, phytochemical properties and antioxidant activity of Coprinus radiata mushrooms was conducted. Free amino acid composition of fresh mushrooms was analyzed by LC-MS/MS. The phytochemical properties analysis included total phenolic compound content and total flavonoid content of these mushrooms. Furthermore, antioxidant activity assays were performed using 2,2-diphenyl-1-picrylhydrazyl (DPPH) and ferric reducing antioxidant power (FRAP). Based on the results shown that a high concentration of non-essential amino acids, especially those of arginine and glutamic acid, were observed. However, isoleucine and leucine were major essential amino acids in these mushrooms. The ethanolic extract of C. radiata showed higher phytochemical contents and antioxidant activity than those of the water extract. It suggested that the C. radiata is a natural source containing some essential amino acids, a high amount of phytochemical properties and high antioxidant activity, potentially having medicinal and nutritional properties that provide health benefits.
MlrA is a gene involved in the degradation of toxic cyanobacterial microcystins. This gene encodes microcystinase, mlrA, the 1st enzyme in the pathway that breaks down toxic cyanobacterial microcystins. In this study, primers were designed, and polymerase chain reaction (PCR) was performed to amplify the mlrA gene in Novosphincgobium sp. KKU25s. A PCR product of 752 base pairs was obtained. The nucleotide sequence of the mlrA gene of Novosphingobium sp. KKU25s was similar to that of Sphingomonas sp. ACM-3962 (98 % similarity). The mlrA gene of Novosphingobium sp. KKU25s was further cloned into the pGEM T-Easy plasmid to obtain the nucleotide sequence of the mlrA gene. The gene was also ligated into the pET32a plasmid for gene expression. Expression was induced by isopropyl β-D-1-thiogalactopyranoside (IPTG) and verified using SDS-PAGE. The expressed protein was approximately 22 kilodaltons. The cell-free extract (CE) containing the crude protein from confirmed recombinant cells showed high activity in the biodegradation of [Dha7] MC-LR. [Dha7] MC-LR at an initial concentration of 30 mg L-1 and was completely biodegraded within 30 h. A distinct product derived from [Dha7] MC-LR appeared with a decrease in the [Dha7] MC-LR peak in the HPLC profile. The product (m/z 999.51) showed a molecular weight of 18, which is higher than that of native [Dha7] MC-LR (m/z 981.50), and was determined to be a linearized peptide fragment of [Dha7] MC-LR using LC-MS analysis. The 3-dimensional structure of microcystinase was predicted from the amino acid sequence deduced from the mlrA gene by the Swiss Model and Phyre2 programs. The structure contained a predicted alpha helix. The predicted 3-dimensional structure was also similar to that of a protein in the CAAX protease group. HIGHLIGHTS Research focused on characterization of microcystinase (MlrA) protein First research worked on the degradation of [Dha7] MC-LR by MlrA This work is useful for the applications aimed at the removal of MCs in freshwater environments GRAPHICAL ABSTRACT
BACKGROUND: Substance use disorders (SUDs) are one of the most stigmatized health conditions that impact drug user's treatment engagement. However, to date, little is known about perceived stigma and its correlates with psychological factors among high-risk groups such as people with SUDs in the criminal justice system. This study aimed to determine the association of psychological factors and perceived stigma of addiction among women with SUDs. MATERIALS AND METHODS: This cross-sectional design was conducted on 652 women with SUDs who were treated in all 7 compulsory drug detention centers in Thailand with consecutive sampling technique. The data were collected by standardized interviewers with interviewing questionnaire. Multiple logistic regression was applied to examine the effect of psychological factors and perceived stigma. RESULTS: More than half of SUD patients (57.2%) were methamphetamine abuse or dependence, approximately 69.9% reported high level of perceived stigma, 56.7% had mild depression, 34.8% had low level of perceived social support, and the average perceived stress score was 19.2 (standard deviation, 7.5). The perceived stigma was positively related to depressive symptom and perceived stress while was inversely related to perceived social support. CONCLUSION: The psychological factors were strongly associated with perceived stigma of addiction. Thus, the detection of psychological problems among SUD patients may be benefit clinicians for identifying which patients are most at risk of perceived stigma and are the potential targets of intervention to reduce stigma in clinical practice.
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