Normal sperm head morphometric reference values in fertile Asian males

Sperm head morphology is crucial for male factor infertility diagnosis and assessment of male reproductive potential.Severalcriteria are available to analyze sperm head morphology,but they are limited by poor methodology comparability and populationapplicability.This study aimed to explore comprehensive and new normal morphometric reference values for spermatozoa heads infertile Asian males.An automated sperm morphology analysis system captured 23152 stained spermatozoa from confirmed fertilemales.Of these samples,1856 sperm head images were annotated by three experienced laboratory technicians as“normal”.Weemployed 14 novel morphometric features to describe sperm head size(head length,head width,length/width ratio,and girth),shape(ellipse intersection over union,girth intersection over union,short-axis symmetry,and long-axis symmetry),area(head,acrosome,postacrosomal areas,and acrosome area ratio),and degrees of acrosome and nuclear uniformity.This straight-forwardmethod for the morphometric analysis of sperm by accurate visual measurements is clinically applicable.The measured parameterspresent valuable information to establish morphometric reference intervals for normal sperm heads in fertile Asian males.Thepresented detailed measurement data will be valuable for interlaboratory comparisons and technician training.In vitro fertilizationand andrology laboratory technicians can use these parameters to perform objective morphology evaluation when assessing malefertilization potential.

SCNN1B and CA12 play vital roles in occurrence of congenital bilateral absence of vas deferens(CBAVD)

Dear Editor,Congenital bilateral absence of the vas deferens(CBAVD),a complete or partial defect of the Wolffian duct derivatives,is found in>25% of men with obstructive azoospermia(OA),but the underlying pathological mechanism remains poorly understood.Previous research has shown that the most common disease associated with CBAVD is cystic fibrosis(CF),the predominant manifestations of which include progressive lung disease,pancreatic dysfunction,elevated sweat chloride electrolyte,meconium ileus,and male infertility.

Supplementation with L-Carnitine Rescues Sperm Epigenetic Changes in Asthenospermic Male Semen with Altered Acetyl-L-Carnitine Levels

Objective:To investigate the relationship between the concentration of L-carnitine in semen and sperm parameters and investigate the epigenetic profile in sperm cell after L-carnitine usage.Methods:From February 2017 to February 2018,46 semen samples from asthenospermic males and 41 semen samples from healthy donors were acquired.Motility parameters were assessed using computer-assisted sperm analysis(CASA,n=78)and the DNA fragmentation index(DFI)was evaluated through flow cytometry(n=86),%DFI=%cells outside main population.Other oxidative stress markers,such as reactive oxygen species(ROS)levels(n=86)and the mitochondria DNA copy numbers,were detected(n=78).The concentration of L-carnitine and acetyl-L-carnitine was detected(n=82),and methylation was analyzed(n=30).After that,we collected 13 fresh semen samples from asthenospermic males and 23 fresh semen samples from healthy donors.These samples were used in a freeze-thaw model that was used to determine whether adding L-carnitine could change sperm progressive motility(n=23),apoptosis index(n=9),and methylation analysis(n=7).In total,we have done 13 asthenospermia samples for Western blot,and except for the poor Western result,we analyzed 6 samples for H3K9ac detection,7 samples for H3K9m3 and H3K27m3 detection,and immunofluorescence(n=3).Finally,we had recruited 30 volunteers,and they were given oral administration of L-carnitine for 3 months and then collected semen samples at different time points for methylation analysis.Results:The concentration of acetyl-L-carnitine is negatively correlated with the%DFI value(r^2=0.1090;P=0.0026),and the concentration of acetyl-L-carnitine is positively correlated with sperm forward motility(r^2=0.0543;P=0.0458)and ROS(r^2=0.1854;P<0.0001),and the acetyl-L-carnitine level is negatively correlated with%DFI in asthenospermia(r^2=0.1701;P=0.0066),and the level of acetyl-L-carnitine in asthenospermic semen is significantly lower than the normal group(P=0.0419).In addition,this study indicates that adding L-carnitine significantly improved sperm motility(P=0.0325)and reduced sperm apoptosis(P=0.0032).Importantly,Western blotting(P=0.0429)and immunofluorescence staining results showed that the addition of L-carnitine reduced H3K9Me3 methylation level in sperm,respectively.Furthermore,semen samples from asthenospermic patients had reduced methylation levels in a specific region(16^thP=0.0003;17^thP=0.0016)of the brain-derived neurotrophic factor(BDNF)promoter.The 16^th methylation decreased with age(r^2=0.1564;P=0.0306),and the 17^th methylation was decreased after treatment with L-carnitine for 28 days(P=0.0341).Conclusion:L-carnitine can reduce the%DFI and also affect the methylation of the histone modification marker in sperm as a possible epigenetic regulator.