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Analyzing all biological evidence at a crime scene presents serious time, budget, and labor constraints. Therefore, selecting valid evidence is crucial for efficient screening. The ABO blood group is a marker that can serve as valid evidence for identifying investigative leads in criminal case. Conventional identification of ABO blood groups using serological methods has only been for blood and is difficult to apply to other body fluids. ABO genotyping was conducted by analyzing single nucleotide polymorphisms (SNP) representative of each blood group. However, this method is time-consuming, expensive, and requires sophisticated instruments. In this study, we developed rapid ABO genotyping method using loop-mediated isothermal amplification (LAMP) and multiplex real-time polymerase chain reaction (PCR). Three SNP sites in the ABO gene (nt 261, 526, and 803) were selected to classify the ABO genotypes. For the specificity test, we performed sequencing of 60 saliva samples to confirm that the genotyping. We conducted experiments to apply ABO genotyping using two amplification methods to mock forensic sample using cotton swab and filter paper. As a result, using LAMP, we successfully identified six ABO genotypes within 30 min at a constant temperature (65 â). Moreover, by using multiple real-time PCR, it was possible to detect not only the major group but also the subgroup of the ABO genotype (ex. cis-AB). The amplification results using the new methods were in concordance with the sequencing results. Therefore, these ABO genotyping methods are expected to select valid evidence successfully and efficiently at the crime scene.
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Técnicas de Genotipaje , Técnicas de Diagnóstico Molecular , Técnicas de Amplificación de Ácido Nucleico , Humanos , Reacción en Cadena en Tiempo Real de la Polimerasa/métodos , Genotipo , Técnicas de Genotipaje/métodos , Sistema del Grupo Sanguíneo ABO/genética , Sensibilidad y EspecificidadRESUMEN
This study evaluated the positive effects of autumn olive berries (AOBs) extract on delaying aging by improving lipid metabolism in middle-aged Caenorhabditis elegans that had become obese due to a high-glucose (GLU) diet. The total phenolic content and DPPH radical scavenging abilities of freeze-dried AOBs (FAOBs) or spray-dried AOBs (SAOBs) were examined, and FAOBs exhibited better antioxidant activity. HPLC analysis confirmed that catechin is the main phenolic compound of AOBs; its content was 5.95 times higher in FAOBs than in SAOBs. Therefore, FAOBs were used in subsequent in vivo experiments. FAOBs inhibited lipid accumulation in both the young adult and middle-aged groups in a concentration-dependent manner under both normal and 2% GLU conditions. Additionally, FAOBs inhibited ROS accumulation in a concentration-dependent manner under normal and 2% GLU conditions in the middle-aged worms. In particular, FAOB also increased body bending and egg production in middle-aged worms. To confirm the intervention of genetic factors related to lipid metabolism from the effects of FAOB, body lipid accumulation was confirmed using worms deficient in the daf-16, atgl-1, aak-1, and akt-1 genes. Regarding the effect of FAOB on reducing lipid accumulation, the impact was nullified in daf-16-deficient worms under the 2% GLU condition, and nullified in both the daf-16- and atgl-1-deficient worms under fasting conditions. In conclusion, FAOB mediated daf-16 and atgl-1 to regulate lipogenesis and lipolysis in middle-aged worms. Our findings suggest that FAOB improves lipid metabolism in metabolically impaired middle-aged worms, contributing to its age-delaying effect.
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Proteínas de Caenorhabditis elegans , Elaeagnaceae , Olea , Animales , Caenorhabditis elegans/metabolismo , Metabolismo de los Lípidos , Olea/metabolismo , Proteínas de Caenorhabditis elegans/genética , Proteínas de Caenorhabditis elegans/metabolismo , Frutas/metabolismo , Envejecimiento , Elaeagnaceae/metabolismo , Lípidos/farmacología , LongevidadRESUMEN
INTRODUCTION: Exposure to electronic cigarette (EC) marketing is associated with EC use, particularly among youth. In England, the Tobacco and Related Products Regulations and Committee of Advertising Practice (CAP) regulate EC marketing to reduce appeal to youth; however, there are little published data on EC marketing claims used online. This study therefore provides an overview of marketing claims present on the websites of EC brands popular in England. METHODS: From January to February 2022, a content analysis of 10 of England's most popular EC brand websites was conducted, including violation of CAP codes. RESULTS: Of the 10 websites, all presented ECs as an alternative to smoking, 8 as a smoking cessation aid and 6 as less harmful than smoking. Four websites presented ECs as risk-free. All mentioned product quality, modernity, convenience, sensory experiences and vendor promotions. Nine featured claims about flavours, colours, customisability and nicotine salts. Seven featured claims concerning social benefits, personal identity, sustainability, secondhand smoke and nicotine strength. Six featured claims about fire safety. Some claimed ECs are cheaper than tobacco (n=5), cited health professionals (n=4) or featured collaborations with brands/icons (n=4). All were assessed by the research team to violate one or more CAP code(s) by featuring medicinal claims (n=8), contents which may appeal to non-smokers (n=7), associations with youth culture (n=6), depictions of youth using ECs (n=6) or media targeting youth (n=5). CONCLUSION: Among 10 top EC brand websites in England, marketing elements that might appeal to youth were commonly identified and CAP code compliance was low.
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To evaluate the value of Cirsium japonicum (CJ; thistle) as a material for functional foods, we studied the functional composition of cultivated CJ and the in vitro and in vivo antioxidant activity of the functional substance. The detected phenolics in farmed CJ were chlorogenic acid (CA), linarin (LIN), and pectolinarin (PLIN) by HPLC analysis. As a result of the antioxidant activity of CJ and its phenolics by DPPH and ABTS method, CA had shown the greatest antioxidant activity. We employed Caenorhabditis elegans to validate that in vitro effects of CA are shown in vivo. CA delayed reduction in pumping rate and progeny production during aging of C. elegans. Under both normal and oxidative stress conditions, CA reduced the production of reactive oxygen species (ROS) in worms and increased their lifespan. In particular, CA showed the reducing effect of ROS accumulation due to aging in aged worms (8 days old). To gain insight into the mechanism, we used skn-1/Nrf2 and daf-16/FOXO transformed worms. The CA effects (on catalase activity and lifespan extension) in the wild-type (WT) decreased in skn-1 and daf-16 mutants. In particular, CA strongly relied on daf-16 under mild oxidative condition and skn-1 under overall (from mild to strong) oxidative stress to reduce ROS and extend healthspan. Thus, we conclude that CA, a key bioactive phenolic of CJ, reduces ROS production and ultimately extends healthspan, and this effect is the result of actions of daf-16 or skn-1 at different stages depending on the degree of oxidation or aging. Our results suggest that CJ containing CA can be used as an antiaging material due to its antioxidant properties.
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We aimed to investigate the risk factors of early double-J ureteral stent (DJUS) dysfunction rates and the long-term patency of DJUSs inserted via a percutaneous approach in patients with benign uretero-ileal anastomosis stricture (UIAS) who had undergone radical cystectomy. In this retrospective study, 63 DJUS placements were placed via a percutaneous nephrostomy tract in 42 consecutive patients between May 2020 and March 2023. The technical success rate was 100% in all patients without major complications. The early dysfunction rate and long-term patency rate were 38.1% (24/63) and 84.2% (32/38), respectively. The blood clot retention grade, balloon dilatation, and length of the ureteral stricture exhibited a significant correlation with early DJUS dysfunction (blood clot retention grade: odds ratio (OR) 6.922 in grade two, p = 0.009; balloon dilatation: OR 0.186, p = 0.017; length of ureteral stricture: OR 8.715, p = 0.035 in moderate stenosis, and 7.646, p = 0.028 in severe stenosis). A multivariate Cox's proportional hazard analysis revealed that blood clot retention grade and length of ureteral stricture were independent predictors of long-term DJUS patency. Percutaneous insertion of the DJUSs was safe and effective in patients with benign UIAS.
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This study aimed to evaluate the antioxidant and antiaging effects of Indian almond (Terminalia catappa Linn.) leaf extract (TCE) on high-glucose (GLU)-induced obese Caenorhabditis elegans. Since TCE contains high contents of flavonoids and phenolics, strong radical scavenging activity was confirmed in vitro. The stress-resistance effect of TCE was confirmed under thermal and oxidative stress conditions at nontoxic tested concentrations (6.25, 12.5, and 25 µg/mL). GLU at 2% caused lipid and reactive oxygen species (ROS) accumulation in C. elegans, and TCE inhibited lipid and ROS accumulation under both normal and 2% GLU conditions in a concentration-dependent manner. In addition, TCE proved to be effective in prolonging the lifespan of C. elegans under normal and 2% GLU conditions. The ROS reduction effect of TCE was abolished in mutants deficient in daf-16/FOXO and skn-1/Nrf-2. In addition, the lifespan-extending effect of TCE in these two mutants disappeared. The lifespan-extending effect was abolished even in atgl-1/ATGL-deficiency mutants. The TCE effect was reduced in aak-1/AMPK-deficient mutants and completely abolished under 2% GLU conditions. Therefore, the effect of prolonging lifespan by inhibiting lipid and ROS accumulation under the high GLU conditions of TCE is considered to be the result of atgl-1, daf-16, and skn-1 being downregulated by aak-1. These results suggest that the physiological potential of TCE contributes to antiaging under metabolic disorders.
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Polystyrene (PS), a major plastic waste, is difficult to biodegrade due to its unique chemical structure that comprises phenyl moieties attached to long linear alkanes. In this study, we investigated the biodegradation of PS by mesophilic bacterial cultures obtained from various soils in common environments. Two new strains, Pseudomonas lini JNU01 and Acinetobacter johnsonii JNU01, were specifically enriched in non-carbonaceous nutrient medium, with PS as the only source of carbon. Their growth after culturing in basal media increased more than 3-fold in the presence of PS. Fourier transform infrared spectroscopy analysis, used to confirm the formation of hydroxyl groups and potentially additional chemical bond groups, showed an increase in the amount of oxidized PS samples. Moreover, field emission scanning electron microcopy analysis confirmed PS biodegradation by biofilms of the screened microbes. Water contact angle measurement additionally offered insights into the increased hydrophilic characteristics of PS films. Bioinformatics and transcriptional analysis of A. johnsonii JNU01 revealed alkane-1-monooxygenase (AlkB) to be involved in PS biodegradation, which was confirmed by the hydroxylation of PS using recombinant AlkB. These results provide significant insights into the discovery of novel functions of Pseudomonas sp. and Acinetobacter sp., as well as their potential as PS decomposers.