Composition and Acute Inflammatory Response from Tetraponera rufonigra Venom on RAW 264.7 Macrophage Cells

Tetraponera rufonigra (Arboreal Bicoloured Ant) venom induces pain, inflammation, and anaphylaxis in people and has an increased incident in Southeast Asia regions. The bioactive components and mechanism of action of the ant venom are still limited. The aim of this research was to identify the protein composition and inflammatory process of the ant venom by using RAW 264.7 macrophage cells. The major venom proteins are composed of 5’ nucleotidase, prolyl endopeptidase-like, aminopeptidase N, trypsin-3, venom protein, and phospholipase A2 (PLA2). The venom showed PLA2 activity and represented 0.46 μg of PLA2 bee venom equivalent/μg crude venom protein. The venom induced cytotoxic in a dose- and time-dependent manner with IC20 approximately at 4.01 µg/mL. The increased levels of COX-2 and PGE2 were observed after 1 h of treatment correlating with an upregulation of COX-2 expression. Moreover, the level of mPGES-1 expression was obviously increased after 12 h of venom induction. Hence, our results suggested that the induction of COX-2/mPGEs-1 pathway could be a direct pathway for the ant venom-induced inflammation

Effect of <i>Nelumbo nucifera</i> Petals Extract on Antioxidant Activity and Sperm Quality in Charolais Cattle Sperm Induced by Mancozeb

The white Nelumbo nucifera petals aqueous extraction (NAE) was prominent in phytochemical content, antioxidant activity, and enhanced rat sperm viability induced by FeSO4, a heavy metal. Mancozeb (MZ) contains heavy metals and is widely used for fungal control in agriculture and industry. It induces oxidative stress and causes of spermatogenesis and reproductive organs’ abnormalities in both humans and animals. The aims of the present study were to investigate the effects of white Nelumbo nucifera petals aqueous extraction (WNAE) on sperm quality in cattle sperm induced by MZ. Moreover, this study investigated phytochemical compounds by liquid chromatography-mass spectrometry. A protein profile related to sperm quality with SDS-page and sperm energy preservation for each treatment was determined. The results found nine phytochemical compounds, in which quercetin-3-O-arabinoglycoside was a major flavonoid that was found in the WNAE. MZ induced free radicals in cells, leading to LPO and protein oxidation, while decreasing sperm motility, sperm viability, acrosome integrity, and normal sperm morphology. The cattle sperm found four proteins related to sperm quality including MWs of 17, 31, 34, and 55 kDa. The WNAE effectively increased energy preservation, sperm motility, sperm viability, acrosome integrity, and normal sperm morphology. The WNAE enhanced sperm qualities by reducing oxidative stress. It might be suggested that WNAE has benefits for sperm preservation which may be used to guard against toxicity in animals or humans exposed to MZ contaminants

Phytochemical Screening, Antioxidant and Sperm Viability of Nelumbo nucifera Petal Extracts

Sacred lotus (Nelumbo nucifera Gaertn.; N. nucifera) is a common ingredient in traditional medicine and Thai recipes. Its petal is an agricultural waste from stamen production. There are limitations in the used and pharmacological data of the petals resulting in more petals waste. The aims of this study were to investigate the phytochemical contents, antioxidant activity, and potential effects on sperm viability of aqueous (NAE) and ethanolic extracts (NEE) of both red and white N. nucifera petals. The white NAE had the highest total phenolics content, total tannins content and maximal antioxidant activity. The white NEE had the highest concentration of total flavonoids. Quercetin was a major flavonoid and was found in the aqueous extracts. Both red and white of NAE in the range of 0.22 to 1.76 mg/mL increased sperm viability. The white NAE was prominent in phytochemical content, antioxidant activity, and both red and white NAE effectively increased rat sperm viability in the in vitro model. The white NAE enhanced sperm viability by decreasing oxidative stress. It might be suggested that the N. nucifera petals have benefits for sperm viability health promotion and may increase the economic value of agricultural waste

Categorization of Cytochrome P4502D6 Activity Score by Urinary Amphetamine/Methamphetamine Ratios

Methamphetamine (MA) level in urine has been used for judgment in MA consumption. Metabolism and intoxication of MA are correlated with the activity of cytochrome P450 2D6 (CYP2D6). The activity score (AS) is a potential tool for predicting exposure and personalized dose of drugs metabolized by CYP2D6. Prediction of the CYP2D6 activity score might be described as MA intoxication. The objective of this study was to categorize the CYP2D6 activity score using the urinary amphetamine (AM)/MA ratio. Urine samples (n = 23,258) were collected. The levels of MA and AM were determined by a gas chromatography–nitrogen–phosphorus detector. The log AS was calculated by an AM/MA ratio and classified into four groups following the percentile position: lower than the 2.5th, the 2.5th–the 50th, the 50th–97.5th, and greater than the 97.5th percentile, respectively. The AS value for males presented was less than 0.024, 0.024–0.141, 0.141–0.836, and greater than 0.836. Meanwhile, the AS values were revealed to be lower than 0.023, 0.023–0.148, 0.148–0.850, and higher than 0.850 for females. The AS value of CYP2D6 can be applied to describe the toxicity of MA in forensic crime scenes and relapse behavior

Postmortem Alteration of Purine Metabolism in Coronary Artery Disease

A new approach for assisting in the diagnosis of coronary artery disease (CAD) as a cause of death is essential in cases where complete autopsy examinations are not feasible. The purine pathway has been associated with CAD patients, but the understanding of this pathway in postmortem changes needs to be explored. This study investigated the levels of blood purine metabolites in CAD after death. Heart blood samples (n = 60) were collected and divided into CAD (n = 23) and control groups (n = 37). Purine metabolites were measured via proton nuclear magnetic resonance. Guanosine triphosphate (GTP), nicotinamide adenine dinucleotide (NAD), and xanthine levels significantly decreased (p p < 0.05) in the CAD group compared to the control group. Decreasing xanthine levels may serve as a marker for predicting the cause of death in CAD (AUC = 0.7). Our findings suggest that the purine pathway was interrupted by physiological processes after death, causing the metabolism of the deceased to differ from that of the living. Additionally, xanthine levels should be studied further to better understand their relationship with CAD and used as a biomarker for CAD diagnosis under decomposition and skeletonization settings

Hypoxia and Proton microbeams: Role of Gap Junction Intercellular Communication in Inducing Bystander Responses on Human Lung Cancer Cells and Normal Cells

Hypoxia plays an important role in the development of malignant solid tumor and is associated with therapeutic resistance and poor outcome treatment. This condition may be due to the ability of hypoxic cells to influence bystander response through gap junction intercellular communication (GJIC) that may reduce the effectiveness of radiation therapy (RT) and increase normal tissue toxicity. The present study is aimed to investigate the role of GJIC and hypoxia-induced bystander response in lung cancer (A549) and normal lung fibroblast (WI-38) under normoxia and hypoxia, when exposed to high LET (linear energy transfer) proton microbeams. The results of our study showed that for the first time there was a difference hypoxia-induced bystander response be-tween cancer and normal cells and GJIC plays a key role in the observed effects

Hypoxia and Gap Junction Communication-Induced Bystander Effect

Radiation-induced bystander effect (RIBE) has been identified as an important contributing factor to normal tissue damage and tumor development. However, the RIBE in cancer and normal cells under hypoxia remain unclear. In this study, we investigated the effect of gap junction intercellular communication (GJIC) in confluent A549 lung cancer and WI-38 normal lung cells after exposed to protons microbeam under hypoxic conditions. Following 6 hour incubation, cells were harvested and assayed for colony formation, micronucleus formation, chromosome aberration and western blotting. Our results show that there were differences of RIBE in bystander A549 and WI-38 cells under hypoxia and normoxia. The differences were also observed in the expression of HIF-1 in bystander A549 and WI-38 cells under both conditions. Interestingly, inhibition of GJIC showed a decreased in toxicity of hypoxic bystander A549 cells, but increased in hypoxic bystander WI-38 cells. We demonstrate the first time that GJIC protects bystander normal cells from toxicity while enhancing in bystander cancer cells under hypoxia.日本マイクロビーム生物研究会2021年度シンポジウ

Identification of Microorganisms Dwelling on the 19th Century Lanna Mural Paintings from Northern Thailand Using Culture-Dependent and -Independent Approaches

Lanna painting is a unique type of painting in many temples in the Northern Thai region. Similar to most mural paintings, they usually decay over time partly due to the activity of microbes. This study aimed to investigate the microorganisms from two Lanna masterpiece paintings in two temples that differ in the numbers of visitors using both culture-dependent and -independent approaches. The microorganisms isolated from the murals were also tested for the biodeterioration activities including discoloration, acid production and calcium precipitation. Most microorganisms extracted from the paintings were able to discolor the paints, but only fungi were able to discolor, produce acids and precipitate calcium. The microorganism communities, diversity and functional prediction were also investigated using the culture-independent method. The diversity of microorganisms and functional prediction were different between the two temples. Gammaproteobacteria was the predominant group of bacteria in both temples. However, the fungal communities were different between the two temples as Aspergillus was the most abundant genus in the site with higher number of visitors [Buak Krok Luang temple (BK)]. Conversely, mural paintings at Tha Kham temple (TK) were dominated by the Neodevriesia genera. We noticed that a high number of visitors (Buak Krok Luang) was correlated with microbial contamination from humans while the microbial community at Tha Kham temple had a higher proportion of saprotrophs. These results could be applied to formulate a strategy to mitigate the amount of tourists as well as manage microorganism to slow down the biodeterioration process