Diets containing sea cucumber (Isostichopus badionotus) meals are hypocholesterolemic in young rats

Peer reviewedPublisher PD

Titration-free massively parallel pyrosequencing using trace amounts of starting material

Continuous efforts have been made to improve next-generation sequencing methods for increased robustness and for applications on low amounts of starting material. We applied double-stranded library protocols for the Roche 454 platform to avoid the yield-reducing steps associated with single-stranded library preparation, and applied a highly sensitive Taqman MGB-probe-based quantitative polymerase chain reaction (qPCR) method. The MGB-probe qPCR, which can detect as low as 100 copies, was used to quantify the amount of effective library, i.e. molecules that form functional clones in emulsion PCR. We also demonstrate that the distribution of library molecules on capture beads follows a Poisson distribution. Combining the qPCR and Poisson statistics, the labour-intensive and costly titration can be eliminated and trace amounts of starting material such as precious clinical samples, transcriptomes of small tissue samples and metagenomics on low biomass environments is applicable

Quantitative assessment of heavy metal effects on sperm function using computer‐aided sperm analysis and cytotoxicity assays

One known environmental risk factor impacting on human reproduction is heavy metal pollution. Although some metals (e.g., Cu, Se and Zn) have protective effects on the male reproductive system in low doses, heavy metals can accumulate to toxic levels and result in poor semen quality and decreased sperm function. We investigated the effect of CuSO4 and CdCl2 (10, 50, 100 and 250 μg/ml or 500 μg/ml) on human sperm motility and vitality by using computer‐aided sperm analysis (CASA) and two cytotoxicity assays (WST‐1 and XTT). Several sperm motility parameters were significantly reduced after 5 hr of exposure to the highest concentrations of CuSO4 (250 μg/ml) and CdCl2 (500 μg/ml). The WST‐1 assay also revealed significantly lower absorbance values for 50, 100 and 250 μg/ml CuSO4 and for 500 μg/ml CdCl2; however, no significant effect was seen with XTT. The calculated average IC50 value was 50.31± 4.34 μg/ml for CuSO4 and 392.32 ±76.79 μg/ml for CdCl2. The effects of these metals were confirmed with MgCl2, a positive control. This study provides threshold concentrations for the harmful effect of CuSO4 and CdCl2 on human spermatozoa and recommends the use of WST‐1 as vitality assay in future in vitro studies

Assessment of vitamin D intake among Libyan women - adaptation and validation of specific food frequency questionnaire

Vitamin D deficiency (VDD) has pandemic proportions worldwide. Numerous studies report on high prevalence of VDD in sunny regions like Near East and North Africa (NENA). Previous studies indicated that Libyan population was at risk of VDD. To contribute to the body of evidence, measurement of vitamin D status on children, adults, in Misurata region was conducted, and confirmed with validated dietary intake study. Serum 25(OH)D was analysed using electrochemiluminescence protein binding assay. Existing Food Frequency Questionnaires (FFQ) were adapted to Libyan Women Food Frequency Questionnaire (LW-FFQ). Repeated 24h dietary recalls and LW-FFQ were employed in vitamin D intake evaluation. LW-FFQ was validated using 24h dietary recall and vitamin D status as referent methods. The questionnaires included anthropometry and lifestyle information. Vitamin D status assessment revealed inadequate levels (25(OH)D lt 50nmol/l) in almost 80% of participants. Women (25-64y) were identified as the most vulnerable group with vitamin D inadequacy present in 82% (61.6% had 25(OH)D lt 25nmol/l, and 20.2% had 25-50nmol/l 25(OH)D). Average Vitamin D intake within the study sample (n=316) was 3.9 +/- 7.9 mu g/d, with 92% participants below both Institute of Medicine (IOM) (10 mu g/d) and European Food Safety Authority (15 mu g/d) recommendations. Measured vitamin D status, in 13% of this group, correlated significantly (p=0.015) with intake estimates. Based on self-report, consumption of vitamin D supplements does not exist among study participants. Additional lifestyle factors influencing vitamin D status were analysed. Only 2% of study participants spend approximately 11 min on the sun daily, 60.4% were obese, 23.1% were overweight and 71.2% reported low physical activity. These findings confirm previous reports on high prevalence of VDD in women across NENA, and in Libya. The situation calls for multi-sectoral actions and public health initiatives to address dietary and lifestyle habits