Neutral Α-glucosidase Research Articles

Activating Transcription Factor 6 Mediates Inflammation in Experimental Varicocele-Induced Epididymal Epithelial Cells.

Varicocele is a dilatation of the internal spermatic vein and it is generally recognized as one cause of male infertility. This study aimed to analyze the roles of activating transcription factor 6 (ATF-6) in experimental varicocele-induced epididymal epithelial cells. Experimental left varicocele was established in rats through partial left renal vein ligation. At 8weeks after surgery, the left epididymal damage was observed using H&E and TUNEL staining. The expressions of neutral α-glucosidase (NAG), ATF-6, tumor necrosis factor (TNF)-α, and phospho-nuclear factor (p-NF)-κB p65 (S536) in the left epididymis were measured by immunohistochemistry. ATF-6 silence in rat epididymal epithelial cells was established by ATF-6 siRNA transfection. The cells were treated with hypoxia for 24h, and cell viability was measured by CCK-8, levels of NAG, TNF-α, and interleukin (IL)-8 in cells were measured by ELISA, levels of p-NF-κB p65 (S536)/NF-κB p65 protein in cells were measured by Western blotting. The results showed that the experimental left varicocele induced hypertrophy and apoptosis of epididymal epithelial cells (p<0.05), and decreased the expressions of NAG in the epididymal epithelial cells compared with the sham-operated control rats (p<0.01). Meanwhile, the expressions of ATF-6, TNF-α, and p-NF-κB p65 (S536) were increased in the epididymal epithelial cells after the experimental left varicocele compared with the sham-operated control rats (p<0.05). In the hypoxia-treated cells, ATF-6 silence increased the cell viability and decreased the levels of TNF-α, IL-8, and p-NF-κB p65 (S536) compared with the control cells (p<0.05). The ATF-6 pathway was activated in a rat's left varicocele-induced epididymal damage. Inhibition of the ATF-6 pathway might be a possible novel therapeutic approach for left varicocele-induced epididymal damage.

The Effect of Neutral Alpha-Glucosidase on Semen Parameters.

The objective of this study was to investigate the relationship between the activity of neutral α-glucosidase in seminal plasma and semen quality and to explore the effect of secretory capability of the epididymis on male fertility. A retrospective analysis of 542 men treated in the Center for Reproductive Medicine and Infertility from February to December 2022, the semen parameters and neutral α-glucosidase were tested and compared among different groups. These 542 men included normozoospermia, oligospermia, asthenospermia, and teratozoospermia. There was statistical difference in neutral alpha-glucosidase (NAG) level among different groups with different sperm concentration, motility, and morphology (p &lt; 0.001). The NAG activity in seminal plasma was positively correlated with ejaculate volume and sperm concentration; meanwhile, a very weak positive correlation was found between NAG level and sperm motility, sperm morphology, respectively. Our results indicated that the secretion of NAG affected the volume, concentration, motility, and morphology of sperm to a certain extent. Given that NAG is a specific and marker enzyme in epididymis, where is the site of sperm maturation, we can conclude that there is a close relationship between NAG and sperm quality. Therefore, seminal plasma NAG has a definite clinical value in helping diagnosis of male infertility.

Establishment and validation of a 5-factor diagnostic model for obstructive and non-obstructive azoospermia based on routine clinical parameters.

Azoospermia is a serious leading male-factor cause of infertility in couples of childbearing age. The two main azoospermia types, obstructive (OA) and non-obstructive (NOA) azoospermia, differ in their treatment approaches. Therefore, their clinical diagnosis is extremely important, requiring an accurate, efficient, and easy-to-use diagnostic model. This retrospective observational study included 707 patients with azoospermia treated between 2017 and 2021, 498 with OA, and 209 with NOA. Hematological and seminal plasma parameters, hormone levels, and testicular volume were used in logistic regression analysis to evaluate and compare their diagnostic performance, results showed that the optimal diagnostic model is constructed by five variables including semen volume, semen pH, seminal plasma neutral α-glucosidase activity, follicle-stimulating hormone in the serum, and testicular volume, compared with follicle-stimulating hormone-based and testicular volume-based models. The 5-factor diagnostic model had an accuracy of 90.4%, sensitivity of 96.4%, positive predictive value of 90.6%, negative predictive value of 89.8%, and area under the curve of 0.931, all higher than in the other two models. However, its specificity (76.1%) was slightly lower than in the other models. Meantime, the internal 5-fold cross-validation results indicated that the 5-factor diagnostic model had a good clinical application value. This study established an accurate, efficient, and relatively accessible 5-factor diagnostic model for OA and NOA, providing a reference for clinical decision-making when selecting an appropriate treatment.

Investigation of lysing excess sludge slurry using hydrolase secreting thermophilic bacterial communities

Sludge reduction is a critical challenge in biological wastewater treatment. Combining excess sludge slurry lysis technology with traditional activated sludge processes is a promising approach for in-situ sludge reduction. Here, a strategy for excess sludge slurry lysis based on thermophilic bacterial communities (LTBC) was proposed. This investigation focused on the process of excess sludge slurry lysis dominated by thermophilic bacterial communities domesticated at different temperatures (55–75 °C). The evolution of sludge lysate was analyzed, and the mechanism of excess sludge slurry lysis under the action of thermophilic bacterial communities was elucidated through amplicon sequencing analysis. The results demonstrated that the aerobic thermophilic bacterial communities adapted to 75 °C exhibit the highest efficiency in sludge slurry lysis. During LTBC process, the removal efficiency of volatile suspended solids reached 53.9 ± 1.8% within 2 h, and 97.0 ± 1.0% of the protein and 96.0 ± 1.0% of the polysaccharide in the extracellular polymers was solubilized, and bacterial cell walls in sludge were disrupted. Fourier transform infrared spectroscopy and excitation-emission matrix spectroscopy of the sludge lysate demonstrated that the LTBC process was accompanied by humification process. The accumulation of humic acid primarily occurred at 55 °C and 65 °C, while fulvic acid occurred at 75 °C. The thermophilic bacterial communities adapted to 75 °C were dominated by Thermus and Thermaerobacter. Phylogenetic studies showed that the LTBC hydrolase system comprises enzymes related to protein hydrolysis, carbohydrate hydrolysis, and peptidoglycan hydrolysis, including metalopeptidase MepB, neutral α-glucosidase C, N-acetyl Muramyl-L-alanine amidase, and others enzymes. These results provide a theoretical basis for the application of LTBC technology in the reduction of sludge which generated in traditional waste water activated sludge processes.

Two fluorimetric determinations of acid α-glucosidase activity in dried blood spot: Pompe disease in Iranian population

IntroductionPompe disease is a lysosomal storage disorder. This study aimed to validate and compare 2 fluorimetric methods for measuring α-glucosidase acid activity in dried blood spot sample (DBS), with potential applications in neonatal screening, and disease follow-up of Pompe patients among the Iranian population for the first time. Materials and methodsThe evaluation involved 3 enzyme levels and 7 parameters. The analysis included 141 Healthy individuals, 8 Pompe patients, and 10 obligate heterozygotes using reference and modified methods. ResultsBoth methods exhibited highly linear calibration curves. The limit of detection (LOD) and limit of quantification (LOQ) were obtained in the micromolar concentration range in 2 methods. Inter-day and intra-day precision, expressed as relative standard deviations (RSD%) were calculated. The normal ranges were determined in healthy individuals. Receiver operating characteristic (ROC) curves were analyzed, and 2 parameters, total neutral α-glucosidase (NAG)/acid α-glucosidase (GAA) and pH ratio, were identified as cut-off values with excellent accuracy, sensitivity, and specificity for evaluating Pompe disease in both methods. ConclusionsEstablishing and implementing these 2 methods for the Iranian population effectively differentiated between healthy and patient individuals. Method II, with its shorter incubation time, demonstrated practicality in the clinical setting.

Cytokine Profile and Enzymes of Tubular Nephrothelium in the Urine of Children with Congenital Megaureter

The objective: to determine the concentration of transforming growth factor β1 (TGF-β1), interleukin 1β (IL-1β) and tumor necrosis factor-α (TNF-α), activity levels of tubular nephrothelium enzymes total N-acetyl-β-D-glucosaminidase (NAG), its thermostable NAG isoenzyme B (NAG-B), β-galactosidase (β-Gal), γ-glutamyl transpeptidase (GGT) and neutral α-glucosidase (Nα-Gl) as cytokine-enzymological risk criteria for the development of nephrofibrosis in the urine of children with congenital nonrefluxing megaureter (NRMU) and refluxing megaureter (RMU) with unilateral and bilateral localization of the pathological process. Materials and methods. 174 patients with congenital MU were examined, who were divided into two groups. Group 1 included 131 children with NRMU, group 2 – 43 children with RMU aged from 6 months to 15 years. The control group included 25 practically healthy children. There were 97 patients with unilateral NRMU (group 1a), with bilateral – 34 persons (group 1b); there were 16 patients with unilateral RMU (group 2a), with bilateral – 27 (group 2b). Cytokine and enzyme indicators were studied in the morning portion of urine before reconstructive treatment. The levels of cytokines were determined by the ELISA method. Enzyme activity of urine was determined by the colorimetric method. Statistical processing of data was performed after analyzing the conformity of their distribution to the norm parameters. Results. It was found that the levels of TGF-β1, IL-1β and TNF-α, as well as the activity levels of NAG, NAG-B, β-Gal, GGT and Nα-Gl in the urine of patients with both congenital NRMU and congenital RMU are significantly increased compared to similar indicators in the control group (p&lt;0.001). In patients with congenital RMU, compared to similar average indicators of patients with congenital NRMU, there is a significant increase of all cytokine and enzyme parameters (р&lt;0.05–0.001), which indicates the probability of ischemic-hypoxic, as well as fibrotic damages in the kidney parenchyma in children with MU, especially in patients with RMU. A significant increase in the cytokine levels and enzyme activity levels were established in patients with NRMU and RMU with a unilateral process compared to similar control data (p&lt;0.001. A statistically significant increase of TNF-α, the activity of NAG-B and β-Gal in patients with bilateral NRMU, the concentration of IL-1β, TNF-α and activity levels of NAG, NAG-B, β-Gal, GGT and Nα-Gl in patients with bilateral RMU compared with similar data in groups of patients with unilateral process was found (p&lt;0.05–0.001). Conclusions. Congenital MU (NRMU, RMU) negatively affects the function of the tubular nephrothelium, which is manifested by an increased level of profibrogenic cytokine TGF-β1 and pro-inflammatory cytokines IL-1β and TNF-α in the urine of children, as well as the activity levels of lysosomal enzymes NAG, NAG-B, β-Gal and brush border enzymes GGT and Nα-Gl, which have renal specific properties. This indicates a high probability of ischemic processes in the kidney parenchyma and the risk of nephrofibrosis development in these patients, especially in patients with RMU with bilateral localization of the pathological process.

RISUG® offers early contraception: An experience during Phase III clinical trials

Objectives: An early contraceptive efficacy with reasonable assurance of reversibility has been a challenge in male contraception. With nearly four decades of research in reversible inhibition of sperm under guidance (RISUG®) as an intravasal male contraceptive, including pre-clinical trials in rats, rabbits, langur monkeys, and three phases of clinical trials, the present study aims to evaluate the additional parameters of a center of Phase III clinical trials. Material and Methods: Subjects were recruited following ICMR guidelines of inclusion and exclusion criteria. Samples were analyzed for sperm functional tests, namely, hypo-osmotic swelling, acrosomal intactness, nuclear chromatin decondensation, and sperm mitochondrial activity index. Furthermore, seminal biochemistry and serum hormones such as follicle-stimulating hormone, luteinizing hormone, testosterone, cortisol, and prolactin were assessed along with levels of anti-sperm antibodies and prostate-specific antigen (PSA). Results: The present study, on human subjects, emphasizes the efficacy of RISUG® with early onset of contraception and indication of a greater possibility of reversal. A significant decrease in all sperm functional parameters was observed following RISUG® injection along with increased sperm abnormalities. Semen biochemistry revealed no marked alterations in the concentration of fructose and acid phosphatase, while significantly decreased levels of glycerophosphorylcholine and neutral α-glucosidase were observed. No significant changes in the circulatory levels of hormones and the levels of PSA were observed. In addition, the development of anti-sperm antibodies, an adverse effect of other vas occlusive methods, was not indicated after RISUG® administration, implying the potential of reversibility in humans as observed earlier in different animal models. Conclusion: RISUG® presenting deleterious effects on spermatozoa and marked alterations in epididymal markers provides early contraception with a greater possibility of reversal. Although the progress of RISUG® toward development as an ideal male contraceptive is slow, the study implies a strong future possibility.

Warming-needle moxibustion with different lengths of moxa stick for asthenospermia with kidney deficiency and liver depression: a randomized controlled trial

To explore the clinical effect of warming-needle moxibustion with different lengths of moxa stick for asthenospermia with kidney deficiency and liver depression. A total of 240 patients with asthenospermia of kidney deficiency and liver depression were randomly divided into a 4-cm group (moxibustion with 4-cm moxa stick, 60 cases, 3 cases dropped off), a 3-cm group (moxibustion with 3-cm moxa stick, 60 cases, 4 cases dropped off), a 2-cm group (moxibustion with 2-cm moxa stick, 60 cases, 2 cases dropped off) and an acupuncture group (60 cases, 3 cases dropped off). All patients were treated with warming-needle moxibustion with different lengths of moxa stick or conventicnal acupuncture at Zhongwan (CV 12), Guanyuan (CV 4), Zhongji (CV 3), Guilai (ST 29), Yaoyangguan (GV 3), Guanyuanshu (BL 26), etc., once a day, five times a week; 4-week treatment was taken as one course, a total of two courses of treatment were given. The semen routine indexes, seminal plasma biochemical indexes, sex hormone levels and TCM syndrome score were compared before and after treatment among the 4 groups. After treatment, the sperm density, sperm viability, ratio of grade A sperm, ratio of grade A and B sperm, seminal plasma fructose and neutral α-glucosidase were higher than those before treatment (P<0.05, P<0.01), and the sperm deformity rates were lower than those before treatment in the 4-cm group and the 3-cm group (P<0.01, P<0.05). The ratio of grade A sperm, ratio of grade A and B sperm, seminal plasma fructose and neutral α-glucosidase in the 4-cm group were higher than the other three groups (P<0.05, P<0.01), and the sperm deformity rate was lower than the other three groups (P<0.05). After treatment, except for dizziness and tinnitus score, each-domain score and total scores of TCM syndrome scale in the 4-cm group and the 3-cm group were lower than those before treatment (P<0.01, P<0.05). The each-domain score of depression, weak waist and knees, low sexual function and total score in the 4-cm group were lower than those in the other three groups (P<0.05, P<0.01). The total effective rate was 87.7% (50/57) in the 4-cm group, which was higher than 78.6% (44/56) in the 3-cm group, 77.6% (45/58) in the 2-cm group and 70.2% (40/57) in the acupuncture group (P<0.05, P<0.01). The warming-needle moxibustion with 4-cm moxa stick could effectively improve quality and motility of sperm and clinical symptoms in patients with asthenospermia of kidney deficiency and liver depression, which is superior to moxibustion with 3-cm, 2-cm moxa sticks and conventional acupuncture.

Peculiarities of Restoration of the Functional Condition of the Kidney Parenchyma in Children with Congenital Defects of the Ureterovesical Segment Depending on Methods of Its Operative Correction (According to Cytokine-enzymological Criteria)

Peculiarities of Restoration of the Functional Condition of the Kidney Parenchyma in Children with Congenital Defects of the Ureterovesical Segment Depending on Methods of Its Operative Correction (According to Cytokine-enzymological Criteria)

Human and animal fertility studies in cystinosis reveal signs of obstructive azoospermia, an altered blood‐testis barrier and a subtherapeutic effect of cysteamine in testis

Cystinosis is an inherited metabolic disorder caused by autosomal recessive mutations in the CTNS gene leading to lysosomal cystine accumulation. The disease primarily affects the kidneys followed by extra‐renal organ involvement later in life. Azoospermia is one of the unclarified complications which are not improved by cysteamine, which is the only available disease‐modifying treatment. We aimed at unraveling the origin of azoospermia in cysteamine‐treated cystinosis by confirming or excluding an obstructive factor, and investigating the effect of cysteamine on fertility in the Ctns −/− mouse model compared with wild type. Azoospermia was present in the vast majority of infantile type cystinosis patients. While spermatogenesis was intact, an enlarged caput epididymis and reduced levels of seminal markers for obstruction neutral α‐glucosidase (NAG) and extracellular matrix protein 1 (ECM1) pointed towards an epididymal obstruction. Histopathological examination in human and mouse testis revealed a disturbed blood‐testis barrier characterized by an altered zonula occludens‐1 (ZO‐1) protein expression. Animal studies ruled out a negative effect of cysteamine on fertility, but showed that cystine accumulation in the testis is irresponsive to regular cysteamine treatment. We conclude that the azoospermia in infantile cystinosis is due to an obstruction related to epididymal dysfunction, irrespective of the severity of an evolving primary hypogonadism. Regular cysteamine treatment does not affect fertility but has subtherapeutic effects on cystine accumulation in testis.

Study of the Functional State of the Proximal Renal Tubules in Patients with Asymptomatic Chronic Heart Failure in Dyslipidemia and its Correction with Simvastatin Treatment

Aim. To study indicators of epithelial dysfunction in the proximal renal tubules by determining the activity of organ-specific enzymes neutral α-glucosidase (NAG) and L-alanine aminopeptidase (LAAP), in patients with the initial stage of chronic heart failure in dyslipidemia, and the possibility of reducing with simvastatin.Material and methods. The study involved 90 subjects, who were divided into control and main groups. The control group consisted of 30 practically healthy individuals, the main group was divided into 2 subgroups: patients with stage I chronic heart failure (CHF) without type 2 diabetes mellitus (DM2) and patients with CHF with DM2. Patients of each of the main subgroups received simvastatin 20-40 mg/day in addition to treatment of the main pathology. The main group included patients with a total serum cholesterol level of more than 6.0 mmol/l, a BMI level of more than 30 kg/m2, and who had not previously taken statins. The exclusion criterion was a violation of the filtration capacity of the kidneys and the presence of gross dysfunction of organs and systems of the body. The functional state of the proximal renal tubules was assessed by the concentration of NAG and LAAP in dialized urine.Results. Initially, the level of activity of renal enzymes in representatives of both major subgroups is higher than the group of practically healthy individuals. Taking simvastatin in the CHF without DM2 subgroup does not cause an increase in enzyme activity throughout the entire observation period, either at a daily dosage of 20 mg (NAG - 12.36±2.65 ncat/1 14.1±5.23 ncat/1 and after 3 and 6 months, LAAP - 9.4±1.62 and 11.2±2.99 ncat/1 after 3 and 6 months), or at a dosage of 40 mg/day (NAG - 30.47±3.85 and 26.2±6.75 ncat/1 after 3 and 6 months; LAAP -17.3±3.56 and 19.58±3.83 ncat/1 after 3 and 6 months). Taking simvastatin 20 mg/day in patients with CHF with DM 2 causes an increase in the NAG activity: 26.68±6.03 and 34.57±9.73 ncat/1 after 3 and 6 months). Taking simvastatin 40 mg/day increase both enzyme activity: NAG -34.3±8.7 and 46.94±9.02 ncat/1 after 3 and 6 months, LAAP - 17.08±5.81 and 22.41±4.89 ncat/1 after 3 and 6 months).Conclusion. The appointment of simvastatin in patients with dyslipidemia on the background of obesity is permissible in order to normalize lipid metabolism. Safe for the functional state of the proximal renal tubules, long-term administration of simvastatin, within the limits of medium-therapeutic dosages, is possible for patients without type 2 diabetes. Long-term use of simvastatin in patients with dyslipidemia on the background of type 2 diabetes mellitus has a negative effect on the epithelium of the proximal renal tubules, in the form of an increase in the activity of renal organ-specific enzymes, which indicates an increased dystrophy of the epithelium.

Сhanges in the functional state of the epithelium of the proximal renal tubules in patients with the initial stage of chronic heart failure during simvastatin therapy.

To assess the change in the functional state of the proximal renal tubule epithelium in patients with dyslipidemia on the background of obesity, by determining the concentration in the urine of the examined level of cystatin C and the degree of activity of the renal organ-specific enzymes neutral α-glucosidase (NAG) and L-alaninaminopeptidase (laap) during simvastatin therapy at a daily dose of 20 mg for 6 months. The study involved 88 people who were divided into three groups: control, comparison and main. The control group is a group of practically healthy individuals: 30 people, average age 20.67 ± 0.18 years, body mass index (BMI) 21.36 ± 0.4 kg/m2. Comparison group (obese): 27 people, average age 22.38 ± 0.76 years, BMI 31.48 ± 0.56 kg / m2. Patients of the main group were divided into 2 subgroups. The first main subgroup of persons with chronic heart failure stage I (CHF I) without type 2 diabetes mellitus (DM 2)) - 15 observed: average age 56.8 ± 1.8 years, BMI 30.28 ± 1.11 kg / m2. The second main subgroup (CHF I with DM 2) - 16 observed: average age 48.25 ± 2.45 years, BMI 30.37 ± 1.11 kg/m2. The study found that simvastatin therapy does not affect glomerular filtration rate in patients with asymptomatic heart dysfunction. There was an increased level of cystatin C in the urine of the comparison group compared to the control group, the concentration of cystatin C in the main subgroups was statistically significantly higher than the control group. On the background of simvastatin therapy for 6 months, the level of this analyte is statistically significantly increased. The activity of LAAP and NAG during simvastatin therapy during the follow-up period in the CHF I subgroup without DM2 significantly decreased. In the subgroup of CHF I + DM2, a decrease in the concentration of LAAP and an increase in the activity of NAG was revealed, which may indicate that the brush border epithelium dystrophy occurred during simvastatin therapy. Simvastatin therapy for 6 months in patients with the initial stage of heart failure at a daily dosage of 20 mg does not impair glomerular function in the form of reduced glomerular filtration rate (GFR). Cystatin C levels are higher in obese individuals without heart failure and significantly higher in those with asymptomatic heart failure. When treating dyslipidemia with simvastatin at a dose of 20 mg / day, there is a decrease in the activity of NAG and laap in patients with CHF I without DM2. In the result of lipid-lowering therapy with simvastatin in a daily dosage of 20 mg in patients with CHF I+D2M there is increased activity of NAG while reducing the concentration of the LAAP, which may be due to degeneration of the proximal tubular epithelium, amid additional load on a partially renal route of metabolism of simvastatin.

A novel laboratory procedure to validate American Urological Association guideline on vasectomy success and to diagnose obstructive azoospermia.

The objective was to develop a laboratory procedure to validate American Urological Association (AUA) Guideline on vasectomy success when nonmotile spermatozoa are found in the post-vasectomy ejaculate. The neutral α-glucosidase (NAG) an epididymal protein assay modified to determine the activity at 30 and 90min of incubation from 24 pre- and 47 post-vasectomy ejaculates. The difference between the two points in the relative activity was calculated and if the difference was nonsignificant will confirm vasectomy success. The mean differences in the relative NAG activity were significantly different in pre- and post-vasectomy ejaculates, respectively. The mean differences in the relative NAG activity were similar in post-vasectomy ejaculates with and without nonmotile spermatozoa. No difference in relative NAG activity in post-vasectomy ejaculates between two time points of incubation may be a reliable method to confirm occlusion of the vas deferens. It also validates the recommendation by AUA Guideline on vasectomy success in the presence of few nonmotile spermatozoa.

The Hydroxyproline Proteome of HeLa Cells with Emphasis on the Active Sites of Protein Disulfide Isomerases.

Hydroxyproline-containing proteins (other than collagens) are rare and difficult to identify. Only 14 such proteins have been found in the human proteome by biochemical methods despite the fact that the list includes examples of biological importance such as hypoxia-inducible factor and the 40S ribosomal protein S23 (RPS23), both of which have significant biological function of the post-translational modification. Comparison of multinotch search software Global-PTM-Discovery to conventional proteomic database search software gave a nine-fold improvement in correctly identifying non-collagen peptides containing hydroxyproline. Manual interpretation of MS-MS spectra refined this list to discover 36 unique peptides representing 24 unique hydroxyproline sites in 21 proteins, of which only Pro62 of RPS23 had been reported previously in UniProt. Eight of the sites were found to be conserved as prolines in nine species examined, ranging from humans to yeast. These include sites 51 and 395 in protein disulfide-isomerase (PDIA1) and sites 204 and 553 in protein disulfide-isomerase A4 (PDIA4). The apparent occupancy of these sites ranged from 72-89%, suggesting a structural and possibly functional role of these PTMs. Fifteen of the sites most likely contain 4R-hydroxyproline (Pro30 of serpin H1, Pro520 of aspartyl/asparaginyl β-hydroxylase, Pro223 of neutral α-glucosidase AB, Pro977 of hypoxia upregulated protein 1, Pro378 of protein ERGIC-53, Pro252 of protein CASC4, Pro545 of bromodomain-containing protein 2, Pro488 of bromodomain-containing protein 3, Pro130 of nucleolar RNA helicase 2, Pro51 of PDIA1, Pro395 of PDIA1, Pro404 of PDIA3, Pro89 of PDIA4, Pro204 of PDIA4, and Pro553 of PDIA4). The remaining sites could be either 4R-hydroxyproline or 3S-hydroxyproline. Recommendations are made to improve automated interpretation of proteomic data to improve future proteomic research whose goal is to mine more of the remaining dark matter of the proteome.

Effects and mechanism of action of transcutaneous electrical acupuncture point stimulation in patients with abnormal semen parameters.

To evaluate the effect of transcutaneous electrical acupuncture point stimulation (TEAS) on sperm parameters and the underlying molecular mechanisms. A total of 121 patients diagnosed with oligozoospermia, asthenozoospermia or oligoasthenozoospermia were randomised into four groups (three treatment groups, one control): the TEAS groups were treated with 2 Hz (n=31), 100 Hz (n=31), or mock stimulation (n=29) at acupuncture points BL23, ST36, CV1 and CV4 for 2 months. The control group (n=30) was provided with lifestyle advice only. The changes in total sperm count and motility in the 2 Hz TEAS group were significantly greater than those in the mock group and the control group. The change in neutral α-glucosidase (NAG) and zinc levels in the 2 Hz group were significantly greater than those in the mock group and control group, and the changes in fructose levels of the 2 Hz group were significantly greater than those in the control group. Significant increases in calcium and integrin-binding protein 1 (CIB1) and reduction of cyclin-dependent kinase 1 b (CDK1) were also found after 2 Hz TEAS treatment. The present findings suggest that 2 Hz TEAS can improve sperm count and motility in patients with abnormal semen parameters, and is associated with increases in seminal plasma zinc, NAG and fructose. The upregulation of CIB1 and downregulation of CDK1 by TEAS may be associated with its positive effects on sperm motility and count. http://www.chictr.org ; registration no. ChiCTR-TRC-11001775.

Improved sperm motility after 4 h of ejaculatory abstinence: role of accessory sex gland secretions.

Various studies have sought to determine the typical v. optimal abstinence period after which semen samples should be collected, with many contradictory results reported. Several factors influence the semen microenvironment, and thus sperm parameters. In this study we focused on the secretions of the prostate, seminal vesicles and the epididymis. Semen samples were obtained from healthy normozoospermic males (n=16) after 4-day and 4-h periods of ejaculatory abstinence, and standard semen analysis was performed using computer-aided sperm analysis, whereas seminal plasma citric acid, neutral α-glucosidase and fructose concentrations were measured using assay kits. There were significant decreases in total sperm count (P<0.001), sperm concentration (P<0.05) and semen volume (P<0.05) after 4h compared with 4 days ejaculatory abstinence. Furthermore, increases were observed in total sperm motility (P<0.05) and sperm progressive motility (P<0.01) after a 4-h abstinence period, accompanied by significant reductions in citric acid (P<0.05), α-glucosidase (P<0.01) and fructose (P<0.01) concentrations. In addition, due to the decreased number of spermatozoa, these concentrations translated to a significant decrease in fructose (P<0.05) per spermatozoon, indicating an intrinsic mechanism capitalising on alternative sources of energy for increased metabolic function and subsequent sperm motility.

Microsurgical Epididymal Cystectomy does not Impact Upon Sperm Count, Motility or Morphology and is a Safe and Effective Treatment for Epididymal Cystic Lesions (ECLs) in Young Men With Fertility Requirements

ObjectiveTo assess the therapeutic efficacy and safety of microsurgical epididymal cystectomy for the treatment of epididymal cystic lesions in young men with fertility requirements and the impact of this surgery upon sperm quality and epididymal function. Materials and MethodsWe prospectively evaluated the therapeutic outcomes, complications, safety and efficacy of microsurgical epididymal cystectomy and the impact of microsurgery upon sperm parameters. All patients were followed-up 3, 6 and 12 months after surgery to investigate sperm count, motility, morphology, neutral α-glucosidase, improvement of symptoms, recurrence, and complications. ResultsPalpated cyst pain was evident in 32 out of 51 patients. Pathologists identified 29 spermatoceles and 22 epididymal cysts. Mean operation time was 39.27 ± 5.98 minutes, and the mean length of postoperative hospital stay was 2.02 ± 0.35 days. After surgery, scrotal pain disappeared in 80.4% of patients (41/51). A significantly higher rate of pain disappearance was observed in patients with palpated pain than those without palpated pain. Insignificant difference was seen in terms of sperm count, motility, sperm morphology or neutral α-glucosidase between preoperation and postoperation at 1-year follow-up. Sperm count and NGA in patients with an epididymal cystic lesions > 2.5 cm were significantly increased after surgery. The rate of complications was 7.3%. No cyst recurrence was observed during follow-up. ConclusionMicrosurgical epididymal cystectomy does not impact upon sperm count, motility, morphology, or epididymal function and is a safe and effective surgical modality for young men with fertility requirements. Local palpated pain on the epididymal cyst is recommended as an optional surgical indication.

Quantitative proteomics and biochemical analyses reveal the role of endoplasmin in the regulation of the expression and secretion of A Disintegrin And Metalloproteinase 12

A Disintegrin And Metalloproteinase 12 (ADAM12) is highly expressed in multiple cancers such as breast and cervical cancers and its high expression reduces the overall patient survival rate. ADAM12 has two major splicing variants, the long membrane-anchored form ADAM12L and the short secreted form ADAM12S. However, how they are regulated and whether they are modulated similarly or differently in cells are not clear. Here, we use affinity purification and mass spectrometry to identify the ADAM12S-interacting proteins. Spectral counting and MaxQuant label-free quantification reveal that ADAM12S but not ADAM12L specifically interacts with a subset of endoplasmic reticulum proteins, such as endoplasmin (GRP94), 78 kDa glucose-regulated protein (GRP78), and UDP-glucose:glycoprotein glucosyltransferase I (UGGT1), that regulate the folding and processing of secreted proteins. Further biochemical experiments validate the interaction between ADAM12S and several of its interacting proteins. Computational docking analysis demonstrates that GRP94 preferentially interacts with ADAM12S over ADAM12L. The data also suggest that both the protein expression level and the secretion of ADAM12S are regulated by GRP94 expression and knockdown. Our results reveal a link between these two proteins that are highly expressed in cancer cells. Furthermore, our studies define a new ADAM12S-specific regulator that may contribute to the cancer development. SignificanceA Disintegrin And Metalloproteinase 12 (ADAM12) is highly expressed in many cancers such as lung, breast, and cervical cancers. ADAM12 has two major splicing variants, the long membrane-anchored form ADAM12L and the short secreted form ADAM12S. However, how they are regulated and whether they are modulated similarly or differently are not completely understood. We use affinity purification and label-free quantitative proteomics to identify the ADAM12S-interacting proteins. Our results reveal that ADAM12S specifically interacts with a subset of endoplasmic reticulum proteins, including endoplasmin (GRP94), UDP-glucose:glycoprotein glucosyltransferase I (UGGT1), and neutral α-glucosidase AB (GANAB). Computer modeling reveals that ADAM12S interacts with the surface amino acids of GRP94 more strongly than ADAM12L. Biochemical experiments further reveal that GRP94 regulates both the protein level and the secretion of ADAM12S. Database mining finds that both GRP94 and ADAM12 are highly expressed in multiple cancers and their high expression is correlated with poor patient survival rate. Taken together, our work discovers a new upstream regulator for ADAM12S, which may contribute to its distinct functions in the regulation of the migration and invasion of cancer cells.

Relationship between asthenozoospermia and selected macroscopic, microscopic and biochemical semen parameters

Asthenozoospermia is a sperm motility disorder in which &lt;32% of spermatozoa show progressive motility, according to the World Health Organization definition (WHO, 2010). Among causes of male infertility asthenozoospermia accounts for nearly 19%. The aim of the study was to determine the relationship between asthenozoospermia and selected macroscopic, microscopic and biochemical parameters of semen. The semen of 112 males from infertile couples was studied. Basic sperm analysis was performed by manual method according to the WHO 2010 guidelines. Macroscopic parameters (volume, pH) and microscopic (total sperm number and concentration, percentage of vital spermatozoa and with normal morphology) were evaluated. Biochemical parameters (neutral α-glucosidase activity – epididymis activity marker, fructose concentration – seminal vesicles activity marker, and citric acid – prostatic activity marker) were evaluated by spectrophotometric method. Patients with asthenozoospermia had a statistically significant decrease in the ejaculate volume, lower total sperm number and lower percentage of vital sperms, as well as lower fructose and citric acid concentrations in comparison to those without asthenozoospermia. There were statistically significant positive correlations between the percentage of spermatozoa showing progressive movement and the ejaculate volume, total number and concentration of spermatozoa, percentage of with normal vitality and morphology, as well as total fructose concentration in the ejaculate. Conclusions: Asthenozoospermia may be associated with abnormal macroscopic and microscopic semen parameters such as reduced ejaculate volume, reduced sperm count, reduced spermatozoa and normal morphology, and reduced biochemical parameters. Co-occurrence of abnormal macroscopic, microscopic and biochemical parameters of semen may indicate a common etiological factor for these disorders.

POST-CHORNOBYL REMOTE RADIATION EFFECTS ON HUMAN SPERM AND SEMINAL PLASMA CHARACTERISTICS

The research was aimed on analysis of the remote consequences of Chornobyl accident on the reproductive function of men adult residing in Ukraine. 232 male volunteers with mean age of 34 years (range 20-47) from 5 different regions of Ukraine (Zhytomyr, Ivano-Frankivsk, Kyiv, Poltava, and Kyiv city) were enrolled in cross-sectional studies of long term radiation effects on seminal plasma and sperm. All manipulations, analysis and classification of ejaculates were done accordingly to WHO recommendations. The content of neutral α-glucosidase, fructose, citric acid in the seminal plasma was determined by spectrophotometry, L-carnitine - by high performance liquid chromatography, zinc - by atomic absorption spectroscopy. In the men residing in the regions heavily contaminated with radioonuclides, the decreased sperm quality with concurrent asthenozoospermia, oligozoospermia, teratozoospermia, asthenoteratozoospermia, oligoasthenoteratozoospermia and oligoteratozoospermia was revealed. Moreover, the concomitant shifts in seminal plasma content of neutral α-glucosidase, fructose, citric acid, L-carnitine and zinc were detected. The study has revealed the ample sperm and semen abnormalities amongst the inhabitants of radiation polluted territories that should have to be a subject of careful research in forthcoming years. This article is a part of a Special Issue entitled "The Chornobyl Nuclear Accident: Thirty Years After".