Food intake diet and sperm characteristics in a blue zone: a Loma Linda Study.

Objectives: The study examined the effect the life-long vegetarian diet on male fertility and focused on vegetarians living in the Loma Linda blue zone, a demographic area known for life longevity. The objective was to compare sperm characteristics of vegetarian with non-vegetarian males.

Study Design: The cross-sectional observational study was based on semen analyses of 474 males from 2009 to 2013.

Further evidence supporting the Diff-Quik stain for sperm chromatin integrity testing.

Objective: To correlate intracytoplasmic sperm injection (ICSI) fertilization with chromatin status assessed by the Diff-Quik procedure modified with a one-minute soak step, and to determine the association of chromatin status with in vitro fertilization (IVF) pregnancy.

Study Design: This was a retrospective study of 81 IVF patients. Gradient-centrifuge washed sperm remaining after ICSI were fixed, stained by Diff-Quik, immersed in water for 1 minute, and analyzed under oil immersion light microscopy.

A simple sperm DNA toroid integrity test and risk of miscarriage.

Current methods of analyzing sperm chromatin competency overlook the inner sperm compartment which is inaccessible to probes and reagents. By breaking the molecular protamine disulfide bridges, the DNA toroids are exposed to integrity analysis. The aim was to develop a simple nuclear toroid test and determine its association with fertilization, pregnancy, and miscarriage.

Addition of eosin to the aniline blue assay to enhance detection of immature sperm histones.

Objectives: To compare the aniline blue assay with and without eosin, and to correlate the results with pregnancy outcome after intracytoplasmic sperm injection (ICSI) procedure.

Design: A retrospective study.

Setting: University-based fertility center.

Retention of membrane charge attributes by cryopreserved-thawed sperm and zeta selection.

Purpose: Mature sperm can be selected based on their negative zeta electrokinetic potential. The zeta selection of cryopreserved sperm is unknown. The objective was to study the effect of zeta processing on the morphology and kinematic parameters of cryopreserved-thawed sperm.

A simple zeta method for sperm selection based on membrane charge.

Objective: The objectives were: [1] to develop a simple zeta potential method for sperm isolation; and [2] to analyze the sperm maturity, morphology, kinematic, and DNA parameters.

Design: The phenomenon of sticky sperm adhering to slide surfaces was adapted for collecting charged sperm.

Setting: Clinical and academic research environment.

Mouse embryonic stem cells for quality control testing in assisted reproductive technology programs.

Objective: To compare the mouse embryonic stem (ES) cell assay with the sperm motility test or 1-cell mouse embryo bioassay for embryotoxic materials.

Study Design: Cryo-preserved-thawed mouse ES-D3 cells, 1-cell mouse embryos and donor sperm were incubated for 1-4 days in culture medium exposed to a control and 4 different test materials. ES cell viability (eosin method), apoptosis (Sybr-Gold fluorescence), development of blastocysts and sperm motility parameters were measured.

Luteal phase serum cell-free DNA as a marker of failed pregnancy after assisted reproductive technology.

Purpose: DNA-damaging factors have been reported in patients that failed to achieve pregnancy after assisted reproductive technologies (ART). The hypothesis was that increased circulating cell-free DNA released by damaged cells could predict unfavorable conditions leading to failed ART treatment. The objective was to compare the relative concentrations of cell-free DNA in the luteal phase sera of nonpregnant versus pregnant patients.

Cyclooxygenase (COX)-2 and granulosa cell apoptosis in vitro.

Purpose: C-myc was studied in cyclooxygenase (COX)-2 associated granulosa cell apoptosis,

Methods: Granulosa cells (N = 5 cases) were incubated for 24 h in either 1 or 50 microM COX-2 inhibitor, 1 or 50 microM COX-1/COX-2 inhibitor, negative or positive controls Single primer polymerase chain reaction of c-myc exon 1 were performed. Bisbenzimide-stained control single-stranded (ssDNA) were hybridized to SYBR Gold-stained ssDNA and fluorescent images analyzed.

Results: C-myc was disrupted by the high-dose COX-2 inhibitor.

Temperature variable and the efficiency of sperm mediated transfection of HPV16 DNA into cells.

Aim: To pretreat sperm at various temperatures before exposure to human papillomavirus (HPV) 16 DNA fragments and to assess the efficiency of HPV carrier sperm to transfect cumulus cells.

Methods: Cumulus cells from follicular aspirates were obtained, pooled and divided into culture dishes containing Sybr Gold-stained HPV DNA carrying sperm that were either pretreated at 4 degree C, 37 degree C or 40 degree C (n = 5). The cells were incubated in 5% CO(2) in air mixture at 37 degree C for 24 hours.

A simple DNA disc chip in a microarray design based on modified comparative genomic hybridization for sperm DNA analysis.

Objective: A DNA disc chip assay, based on comparative genomic hybridization, was designed to measure changes in sperm DNA intensities. The objective was to analyze the DNA integrity of hyperactive sperm cells after mild heat treatment.

Design: The assay based on a multiple cell comet assay was used to analyze changes in genomic DNA.