Semen analysis is characterized by high levels of intra- and inter-laboratory variability, due to a low level of standardization, high subjectivity of the assessments, and problems with automated procedures. To improve consistency of laboratory results, quality control and training of technicians are important requisites. The goals of this study are to evaluate the results of an external quality control (EQC) program and standardized training by ESHRE Basic Semen Analysis Courses (BSAC) on the variability in manual assessments of semen parameters. We performed retrospective analyses of (1) the interlaboratory variability in the Dutch EQC program and (2) the interobserver variability in BSACs for concentration, motility, and morphology assessments. EQC data showed that the interlaboratory coefficient of variation (CV) for concentration assessment decreased (range from 24.0%-97.5% to 12.7%-20.9%) but not for morphology and motility assessments. Concentration variability was lower if improved Neubauer hemocytometers were used. Morphology assessment showed highest CVs (up to 375.0%), with many outliers in the period of 2007-2014. During BSAC, a significant reduction of interobserver variability could be established for all parameters (P < 0.05). The absence of an effect in the EQC program for motility and morphology might be explained by respectively the facts that motility assessment was introduced relatively late in the EQC program (since 2013) and that criteria for morphology assessment changed in time. BSAC results might have been influenced by the pretraining level of participants and the influence of external factors. Both EQC and training show positive effects on reducing variability. Increased willingness by laboratories to change their methods toward standards may lead to further improvements.
Humans ; Netherlands ; Quality Control ; Retrospective Studies ; Semen ; Semen Analysis ; Sperm Count ; Sperm Motility

Advances in the oncology field have led to improved survival rates. Consequently, quality of life after remission is anticipated, which includes the possibility to conceive children. Since cancer treatments are potentially gonadotoxic, fertility preservation must be proposed. Male fertility preservation is mainly based on ejaculated sperm cryopreservation. When this is not possible, testicular sperm extraction (TESE) may be planned. To identify situations in which TESE has been beneficial, a systematic review was conducted. The search was carried out on the PubMed, Scopus, Google Scholar, and CISMeF databases from 1 January 2000 to 19 March 2020. Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) recommendations were followed in selecting items of interest. Thirty-four articles were included in the systematic review, including 15 articles on oncological testicular sperm extraction (oncoTESE), 18 articles on postgonadotoxic treatment TESE and 1 article on both oncoTESE and postgonadotoxic treatment TESE. Testicular sperm freezing was possible for 42.9% to 57.7% of patients before gonadotoxic treatment and for 32.4% to 75.5% of patients after gonadotoxic treatment, depending on the type of malignant disease. Although no formal conclusion could be drawn about the chances to obtain sperm in specific situations, our results suggest that TESE can be proposed before and after gonadotoxic treatment. Before treatment, TESE is more often proposed for men with testicular cancer presenting with azoospermia since TESE can be performed simultaneously with tumor removal or orchiectomy. After chemotherapy, TESE may be planned if the patient presents with persistent azoospermia.
Child ; Humans ; Male ; Azoospermia/therapy* ; Testicular Neoplasms/therapy* ; Quality of Life ; Spermatozoa ; Testis ; Syndrome ; Sperm Retrieval ; Retrospective Studies

Objective: To investigate the application of the self-made semen quality control (QC) product in internal QC of computer-assisted sperm analysis (CASA). METHODS: CASA was calibrated with high- and low-concentration commercially available semen QC product and meanwhile 15 samples of self-made mixed semen QC product were placed in 75 cryotubes containing liquid nitrogen, followed by CASA of the concentration, motility, curvilinear velocity (VCL), straight line velocity (VSL), average path velocity (VAP), linearity (LIN), wobble (WOB) and straightness (STR) of the sperm using standard procedures and 50 days of continuous monitoring. The Makler counting plate was used to measure the concentration and motility of the self-made sperm. RESULTS: The coefficients of variation (CV) of the commercially available semen QC product at high and low concentrations were 6.18% and 7.85%, respectively. CASA showed that the concentration of the self-made QC product was (25.97 ± 1.41) ×10⁶/ml, with a CV of 5.42%, and the sperm motility, VCL, VSL, VAP, LIN, WOB and STR were (22.15 ± 1.75)% (CV = 7.9%), (59.18 ± 2.05) μm/s (CV = 3.46%), (26.79 ± 1.2) μm/s (CV = 4.48%), (34.98 ± 1.4) μm/s (CV = 4.01%), 46.81 ± 1.55 (CV = 3.3%), 60.52 ± 1.3 (CV = 2.15%) and 76.46 ± 1.98 (CV = 2.59%), respectively. The concentration and motility of the self-made sperm detected with the Makler counting plate were (34.39 ± 2.37) ×10⁶/ml (CV = 6.89%) and (38.04 ± 1.69)% (CV = 4.44%), respectively. Levey-Jennings QC charts were plotted for the indicators using the means and standard deviation. CONCLUSIONS The self-made internal QC product by liquid nitrogen cryopreservation is feasible and effective for monitoring the accuracy and precision of CASA-derived sperm concentration and motion parameters, and it has a smaller CV than the commercially available QC product in measuring sperm concentration.
Computers ; Humans ; Male ; Quality Control ; Semen Analysis/standards* ; Sperm Motility ; Spermatozoa

Male infertility is a relatively common condition affecting approximately 1 in 20 of the male population. DNA fragmentation is an important factor in the etiology of male infertility. Men with high DNA fragmentation levels have significantly lower odds of conceiving, naturally or through procedures such as intrauterine insemination and IVF. The most common contributing factor of male infertility is smoking. Studies have shown that smoking intensity is positively associated with job demands and stress. Therefore, we believe that work stress increases the nicotine-dependent thus causing lower male fertility rate. As proper protamine to histone ratio is essential to produce viable sperm, smoking is strongly suspected to reduce sperm viability through histone-to-protamine transition abnormalities. These abnormalities, results in sperm with high DNA damage when exposed to excessive free radical. This present study was undertaken to evaluate the relationship of work stress, smoking and sperm quality. A total of 210 infertile patients attending Medical Assisted Contraceptive Clinic (MAC), UKMMC were selected for the study. Smoking status and stress level of patients were collected after obtaining relevant consent. Histone-to-protamine ratio was acquired using Aniline Blue staining and Chromomycin A3 staining respectively. Sperm DNA fragmentation was estimated using Comet Assay. Result revealed that smokers tend to be more stressful (r = .446, p <. 001). The result showed a significantly increased level of histone (r = .385, p <. 001) and incomplete protamination (r = .492, p <. 001) in smokers. The imbalance of histone-to-protamine ratio lead to increase of DNA damage. All the data were analyzed using SPSS version 20.0. Result revealed that patients who smoke are more stressful at work. Higher proportion of abnormal sperm histone to protamine ratio were found among smokers suggesting that cigarette smoking may inversely affect male fertility.
male ; stress ; smoking ; sperm quality

PURPOSE: Patients with bladder pain syndrome/interstitial cystitis (BPS/IC) can have pain as a main symptom and overactive bladder (OAB) symptoms that are directly or indirectly related to a major mechanism that causes pain. The primary purpose of this study is firstly to identify the prevalence rate of OAB symptoms in patients with BPS/IC, secondly to identify changes in OAB symptoms after low-dose triple therapy, and thirdly to build a theoretical foundation to improve quality of life for patients. METHODS: Patients who met the inclusion criteria of BPS/IC through basic tests including the O'Leary-Sant symptom index, overactive bladder symptom score (OABSS), and visual analog scale (VAS) were identified. Treatment-based changes in OAB symptoms were identified using the IC Symptom Index and IC Problem Index (ICSI/ICPI), OABSS, and VAS before, and 4 and 12 weeks after low-dose triple therapy. RESULTS: The patients consisted of 3 men and 20 women, and their mean age was 61.9 years (41.0-83.2 years). Comparing values before treatment, and 4 and 12 weeks after treatment (baseline vs. 4 weeks to baseline vs. 12 weeks), the rates of improvement were as follows: ICSI, 44.2% to 63.7%; ICPI, 46.9% to 59.4%; OABSS, 34.3% to 58.2%; and VAS, 53.6% to 75.0%, which showed statistically significant differences (P<0.05). However, comparing values at 4 and 12 weeks after treatment (4 weeks vs. 12 weeks), the ICSI and VAS showed a statistically significant decrease (P<0.05). The ICPI and OABSS showed slight improvement, but no statistically significant differences (P>0.05). CONCLUSIONS: Low-dose triple therapy in BPS/IC results in a clear decrease in OAB symptoms in the first 4 weeks after treatment, and additional treatment for 8 weeks had a partial effect with varied statistical significances depending on the questionnaires.
Amines ; Amitriptyline ; Cyclohexanecarboxylic Acids ; Cystitis ; Cystitis, Interstitial ; Female ; gamma-Aminobutyric Acid ; Humans ; Male ; Prevalence ; Quality of Life ; Sperm Injections, Intracytoplasmic ; Urinary Bladder ; Urinary Bladder, Overactive

OBJECTIVETo establish a method for internal quality control (IQC) of sperm concentration test in the laboratory.

METHODSWe set the concentrations of frozen semen at 20 x 10(6) and 80 x 10(6) as low and high concentrations of putative IQC products, with QC-BEADSTM quality control beads (QCBs) as the control. Using the double-blind method, four technicians determined the sperm concentrations of the IQC products and QCBs by computer-assisted sperm analysis, and drew a quality control chart (Xbar chart and Sbar chart) for each product. Through a month of continuous detection, we calculated and compared the intra- and inter-batch coefficients of variation (CV%) of the quality control products of high and low concentrations.

RESULTSThe intra-batch coefficients of variation of the assumed IQC products of high and low concentrations were CV3.5% and CV2.4%, and their inter-batch coefficients of variation were CV10.2% and CV9.6%. The intra-batch coefficients of variation of the QCBs of high and low concentrations were CV5.1% and CV7.1%, and their inter-batch coefficients of variation were CV7.1% and CV8%. The intra-batch coefficients of variation of both IQC products and QCBs of high and low concentrations were <10%, and their inter-batch coefficients of variation were <15%, which conformed to Levey-Jennings quality control principles and achieved IQC purposes. No significant differences were found in either intra- or inter-batch coefficients of variation between the IQC products and QCBs of high and low concentrations (P>0.05), indicating that assumed IQC products can replace QCBs for internal quality control in the laboratory.

CONCLUSIONThe IQC method we established for determining sperm concentration is simple, feasible and reliable.

Double-Blind Method ; Humans ; Male ; Quality Control ; Semen Analysis ; methods ; standards ; Semen Preservation ; Sperm Count ; Sperm Motility ; Spermatozoa

OBJECTIVETo discuss how some pre-analysis processes influence the results of semen analysis and how to minimize their influence on the accuracy of laboratory results based on the concept of total quality management (TQM).

METHODSWe conducted semen quality analyses for 21 male volunteers, who had abstained from tobacco and alcohol for over 72 days for the purpose of fertilization, before and after the abstinence, and obtained their seminal parameters at 0.5, 1, 2 and 3 hours after semen sample collection.

RESULTSSperm concentration, sperm motility and the percentage of grade a + b sperm were significantly higher after the abstinence of tobacco and alcohol than before (P < 0.01). With the lengthening of post-ejaculation time, there was a significant decrease in sperm motility and the percentage of grade a + b sperm (P < 0.05), but not in sperm concentration (P > 0.05).

CONCLUSIONA lot of factors may affect the results of semen analysis, including the subjects' habits of drinking and smoking and the length of time after semen collection. Therefore, every procedure of semen analysis has to be dealt with very carefully so as to meet the requirements of TQM and achieve most reliable results for clinical use.

Adult ; Humans ; Male ; Quality Control ; Semen Analysis ; methods ; standards ; Smoking Cessation ; Sperm Count ; Sperm Motility ; Temperance

OBJECTIVETo investigate and analyze the results of the determination of sperm concentration, fructose concentration, alpha-glucosidase and acid phosphatase (ACP) activities in the seminal plasma from different hospitals in the city of Nanjing, so as to provide a basis for the external quality control (EQC) of semen analysis within Jiangsu Province or even the whole country.

METHODSEight samples of quality control products for low and high concentrations sperm count, fructose, alpha-glucosidase and ACP determination were prepared and divided, each detected for the sperm concentration, fructose, alpha-glucosidase and ACP activity, and the coefficient variances (CVs) were calculated. The products were then distributed to 11 hospitals in the city, and the results were collected and analyzed. In addition, the total relative errors (REs) for each product was calculated based on the results after dividing as reference values.

RESULTSThe CVs from the 8 samples after dividing were 3.83% - 11.16%. Collected from the 11 hospitals attending EQC were 11 reports of the results of sperm concentration, and 5 the results of fructose, alpha-glucosidase and ACP in seminal plasma. Among the results from different laboratories, those of fructose determination showed the minimal difference (CVs: 8.99% and 3.95% for low and high concentrations, respectively) , next came alpha-glucosidase (CVs: 16.66% and 18.41% for low and high activities, respectively), and ACP determination showed the maximal difference (CVs: 54.12% and 65.58% for low and high activities, respectively). Moreover, the same trend was observed in RE values, as shown in the total REs, which were 11.99% (low concentration) and 20.31% (high concentration) for the determination of fructose in seminal plasma, 22.92% and 27.26% for alpha-glucosidase, 7.34% and 318.35% for ACP in different laboratories, and the maximal RE value was detected in the result of the high-activity ACP sample. Of the 11 hospitals, 6 determined sperm concentration with the computer-assisted semen analysis (CASA) system, and the other 5 with the modified hemocytometer. RE values (148.47% and 187.59% for low and high concentration samples, respectively) and sperm concentrations ([62.74 +/- 16.63] x 10(6)/ml and [163.32 +/- 36.24] x 10(6)/ml) counted with the hemocytometer were significantly higher than those with the CASA system (REs 13.97% and 10.48%; sperm concentrations [24.88 +/- 4.16] x 10(6)/ml and [54.24 +/-23.06] x 10(6)/ml ).

CONCLUSIONThe methods of seminal alpha-glucosidase and fructose determination were relatively stable in current andrology laboratories, and the variance range could be accepted. However, the method of seminal ACP determination might be unadaptable to clinical application, and needs to be further improved. Hemocytometer, which significantly overestimated sperm concentration, could not be applied to the assay of sperm concentration.

Acid Phosphatase ; analysis ; China ; Humans ; Male ; Quality Control ; Semen ; enzymology ; Sperm Count ; standards ; Sperm Motility ; alpha-Glucosidases ; analysis

Semen analysis is a basic test to evaluate male reproductive function. In recent years, urgent needs for the standardization of semen analysis have been emphasized among andrologists worldwide. This review discusses the standardization and quality control (QC) for the analysis of sperm quality parameters, including sperm concentration, motility and morphology. The key to sperm concentration analysis is the standardization of sperm-counting chamber, thus Cell-VU chamber may be the first choice. The analysis of sperm motility and morphology is too subjective to be reliable. Therefore, the computer-aided semen analysis (CASA) system may be the final selection. QC of semen analysis mainly lies in the selection of QC materials and the administration of external QC and internal QC. Meanwhile, the charts and arithmetic methods should be established to monitor QC of semen analysis.
Humans ; Male ; Quality Control ; Reference Standards ; Semen ; cytology ; Sperm Count ; instrumentation ; standards ; statistics & numerical data ; Sperm Motility ; Spermatozoa ; cytology ; physiology

The epididymal lumen represents a unique extracellular environment because of the active sperm maturation process that takes place within its confines. Although much focus has been placed on the interaction of epididymal secretory proteins with spermatozoa in the lumen, very little is known regarding how the complex epididymal milieu as a whole is maintained, including mechanisms to prevent or control proteins that may not stay in their native folded state following secretion. Because some misfolded proteins can form cytotoxic aggregate structures known as amyloid, it is likely that control/surveillance mechanisms exist within the epididymis to protect against this process and allow sperm maturation to occur. To study protein aggregation and to identify extracellular quality control mechanisms in the epididymis, we used the cystatin family of cysteine protease inhibitors, including cystatin-related epididymal spermatogenic and cystatin C as molecular models because both proteins have inherent properties to aggregate and form amyloid. In this chapter, we present a brief summary of protein aggregation by the amyloid pathway based on what is known from other organ systems and describe quality control mechanisms that exist intracellularly to control protein misfolding and aggregation. We then present a summary of our studies of cystatin-related epididymal spermatogenic (CRES) oligomerization within the epididymal lumen, including studies suggesting that transglutaminase cross-linking may be one mechanism of extracellular quality control within the epididymis.
Amino Acid Substitution ; Amyloid ; physiology ; standards ; Cystatin C ; Cystatins ; genetics ; Dimerization ; Epididymis ; physiology ; Humans ; Male ; Mutation ; Protein Folding ; Proteins ; standards ; Quality Control ; Sperm Maturation ; physiology ; Transglutaminases ; physiology