Determination of polycyclic aromatic hydrocarbons in beverage by low density solvent based-dispersive liquid-liquid microextraction-high performance liquid chromatography-fluorescence detection

A simple and fast dispersive liquid-liquid microextraction (DLLME) technique using organic solvent lighter than water has been developed for the extraction of selected polycyclic aromatic hydrocarbons in green tea, chrysanthemum tea and coffee beverage. The 1-octanol and acetonitrile were pre-mixed and injected into the sample solution. After dispersing, the cloudy solution was subjected to centrifugation to separate the solution into 2 phases, where the micro-droplet of 1-octanol was clearly floated on the top layer. The 1-octanol extract was then diluted and injected into high performance liquid chromatography-fluorescence for the quantitative analysis. The technique offered rapid analysis as the 1-octanol was homogeneously dispersed in the sample solution thus speeding the analytes diffusion. Under the optimized extraction conditions, the technique achieved trace detection limits in the range of 0.001 to 0.3 μgL-1 for the targeted analytes, namely phenanthrene, fluoranthene and benzo[a]pyrene. The method was successfully applied to the spiked green tea, chrysanthemum tea and coffee beverage samples with good average relative recoveries obtained in the range of 86.7 to 103.0%. The utilization of low density organic solvent as extraction solvent has allowed for easier operation and eliminated the use of hazardous halogenated solvent that is commonly applied in DLLME

Characterization of aldehyde dehydrogenase isozymes in ovarian cancer tissues and sphere cultures

BACKGROUND: Aldehyde dehydrogenases belong to a superfamily of detoxifying enzymes that protect cells from carcinogenic aldehydes. Of the superfamily, ALDH1A1 has gained most attention because current studies have shown that its expression is associated with human cancer stem cells. However, ALDH1A1 is only one of the 19 human ALDH subfamilies currently known. The purpose of the present study was to determine if the expression and activities of other major ALDH isozymes are associated with human ovarian cancer and ovarian cancer sphere cultures. METHODS: Immunohistochemistry was used to delineate ALDH isozyme localization in clinical ovarian tissues. Western Blot analyses were performed on lysates prepared from cancer cell lines and ovarian cancer spheres to confirm the immunohistochemistry findings. Quantitative reverse transcription-polymerase chain reactions were used to measure the mRNA expression levels. The Aldefluor® assay was used to measure ALDH activity in cancer cells from the four tumor subtypes. RESULTS: Immunohistochemical staining showed significant overexpression of ALDH1A3, ALDH3A2, and ALDH7A1 isozymes in ovarian tumors relative to normal ovarian tissues. The expression and activity of ALDH1A1 is tumor type-dependent, as seen from immunohistochemisty, Western blot analysis, and the Aldefluor® assay. The expression was elevated in the mucinous and endometrioid ovarian epithelial tumors than in serous and clear cell tumors. In some serous and most clear cell tumors, ALDH1A1 expression was found in the stromal fibroblasts. RNA expression of all studied ALDH isozymes also showed higher expression in endometrioid and mucinous tumors than in the serous and clear cell subtypes. The expression of ALDH enzymes showed tumor type-dependent induction in ovarian cancer cells growing as sphere suspensions in serum-free medium. CONCLUSIONS: The results of our study indicate that ALDH enzyme expression and activity may be associated with specific cell types in ovarian tumor tissues and vary according to cell states. Elucidating the function of the ALDH isozymes in lineage differentiation and pathogenesis may have significant implications for ovarian cancer pathophysiology

Genome-Wide Association Study of Retinopathy in Individuals without Diabetes

10.1371/journal.pone.0054232PLoS ONE82

Genome-Wide Meta-Analysis of Five Asian Cohorts Identifies PDGFRA as a Susceptibility Locus for Corneal Astigmatism

Corneal astigmatism refers to refractive abnormalities and irregularities in the curvature of the cornea, and this interferes with light being accurately focused at a single point in the eye. This ametropic condition is highly prevalent, influences visual acuity, and is a highly heritable trait. There is currently a paucity of research in the genetic etiology of corneal astigmatism. Here we report the results from five genome-wide association studies of corneal astigmatism across three Asian populations, with an initial discovery set of 4,254 Chinese and Malay individuals consisting of 2,249 cases and 2,005 controls. Replication was obtained from three surveys comprising of 2,139 Indians, an additional 929 Chinese children, and an independent 397 Chinese family trios. Variants in PDGFRA on chromosome 4q12 (lead SNP: rs7677751, allelic odds ratio = 1.26 (95% CI: 1.16–1.36), Pmeta = 7.87×10−9) were identified to be significantly associated with corneal astigmatism, exhibiting consistent effect sizes across all five cohorts. This highlights the potential role of variants in PDGFRA in the genetic etiology of corneal astigmatism across diverse Asian populations

High School Course-Completion Trajectories and College Pathways for All: A Transcript Analysis Study on Elective Computer Science Courses

Whereas researchers regard high school math and science coursework as the best indicator of college readiness for students in the United States, computer science coursework and its relationship to college attendance, particularly for minoritized students, have not received due attention despite its root in the mathematical and scientific reasoning ability. We examined students’ high school course completion patterns across subjects and grade levels with a special focus on elective computer science courses and whether the coursework pattern transitions worked differently for minoritized students in Texas, USA. Latent profile analysis and latent transition analysis revealed multiple patterns of coursework, including Regular, Trailing, and Computer Science-Intensive. However, high school students seemed to attempt computer science courses with an experimental attitude. High school girls, low-income, and Latinx and African American students were less likely to complete computer science courses, despite demonstrating a similar coursework pattern in the previous year. Similarly, students with limited English proficiency, those eligible for free- or reduced-price lunch programs, and Native American students systematically have a lower chance to attend college, despite sufficient academic preparation in high school. Findings highlight the challenges minoritized students face and how students approach elective computer science courses in high school

High School Course-Completion Trajectories and College Pathways for All: A Transcript Analysis Study on Elective Computer Science Courses

Whereas researchers regard high school math and science coursework as the best indicator of college readiness for students in the United States, computer science coursework and its relationship to college attendance, particularly for minoritized students, have not received due attention despite its root in the mathematical and scientific reasoning ability. We examined students’ high school course completion patterns across subjects and grade levels with a special focus on elective computer science courses and whether the coursework pattern transitions worked differently for minoritized students in Texas, USA. Latent profile analysis and latent transition analysis revealed multiple patterns of coursework, including Regular, Trailing, and Computer Science-Intensive. However, high school students seemed to attempt computer science courses with an experimental attitude. High school girls, low-income, and Latinx and African American students were less likely to complete computer science courses, despite demonstrating a similar coursework pattern in the previous year. Similarly, students with limited English proficiency, those eligible for free- or reduced-price lunch programs, and Native American students systematically have a lower chance to attend college, despite sufficient academic preparation in high school. Findings highlight the challenges minoritized students face and how students approach elective computer science courses in high school

Presence of Bacillus coagulans spores and vegetative cells in rat intestine and feces and their physiological effects

This study was aimed to investigate the presence of Bacillus coagulans vegetative cells in the intestine and fecal samples in rats fed B. coagulans spores as well as to estimate the ratios of spores and vegetative cells in these samples. A two-step process has been developed to enumerate B. coagulans in different mixed bacterial samples, specifically (1) observation of yellow ring formation on modified GYEA medium upon incubation at 55°C, (2) microscopic examination of spore formation after 7 d of incubation. Our results have demonstrated the presence of vegetative cells in the intestinal and fecal samples in rats fed B. coagulans spores. The ratios of B. coagulans spores and vegetative cells in cecal fluid, colonic content, and feces were approximately 2:8, 2:8, and 4:6, respectively. The existence of B. coagulans vegetative cells improved the intestinal milieu through an elevated short-chain fatty acid concentrations, higher fecal moisture, and lower fecal pH

Altered striatal dopamine levels in Parkinson's disease VPS35 D620N mutant transgenic aged mice

Vacuolar protein sorting 35 (VPS35) is a major component of the retromer complex that mediates the retrograde transport of cargo proteins from endosomes to the trans-Golgi network. Mutations such as D620N in the VPS35 gene have been identified in patients with autosomal dominant Parkinson's disease (PD). However, it remains poorly understood whether and how VPS35 deficiency or mutation contributes to PD pathogenesis; specifically, the studies that have examined VPS35 thus far have differed in results and methodologies. We generated a VPS35 D620N mouse model using a Rosa26-based transgene expression platform to allow expression in a spatial manner, so as to better address these discrepancies. Here, aged (20-months-old) mice were first subjected to behavioral tests. Subsequently, DAB staining analysis of substantia nigra (SN) dopaminergic neurons with the marker for tyrosine hydroxylase (TH) was performed. Next, HPLC was used to determine dopamine levels, along with levels of its two metabolites, 3,4-dihydroxyphenylacetic acid (DOPAC) and homovanillic acid (HVA), in the striatum. Western blotting was also performed to study the levels of key proteins associated with PD. Lastly, autoradiography (ARG) evaluation of [3H]FE-PE2I binding to the striatal dopamine transporter DAT was carried out. We found that VPS35 D620N Tg mice displayed a significantly higher dopamine level than NTg counterparts. All results were then compared with that of current VPS35 studies to shed light on the disease pathogenesis. Our model allows future studies to explicitly control spatial expression of the transgene which would generate a more reliable PD phenotype.National Medical Research Council (NMRC)Published versionThis research was supported by the Open Fund-Large Collaborative Grant [OFLCG18May-0026]; and Open Fund-Individual Research Grant [NMRC/OFIRG/0074/2018] administered by the Singapore Ministry of Health’s National Medical Research Council