On the basis of the semen analysis in 66 subjects, they were divided into six different groups: Group I consisted of 16 normal subjects with sperm counts of over 40 ${\times}10^6$/ml and motility of over 40 percent, Group II, 7 subjects with normal sperm counts, but motility of under 40 percent, Group III, 15 oligospermic patients with under 40 ${\times}10^6$/ml, Group IV 14 azoospermic patients, Group V, 10 patients with vasectomy and Group VI, 4 abnormal patients with 2 cases of hypoplastic testis, 1 case of Klinefelter's syndrome and 1 case of testis tumor. After seperation of semen into sperm and seminal plasma by centrifugation, the protein contents and the activities of hyaluronidase, ${\beta}$-N acetylglucosaminidase, ${\beta}$-glucuronidase, arylsulfatase, acrosin and azocoll proteinase in seminal plasma were measured. Vasectomy group has 30 percent less of total protein than normal group. For the comparison of enzyme activities of seminal plasma, it could be assumed that the enzymes in seminal plasma were not contaminated with the enzymes of spermatozoa by testing the enzymes of the seminal plasma from the vasectomy and azoospermic groups. It had been reported that hyaluronidase was only released from spermatozoa, however, the result obtained in this investigation showed that azoospermic and vasectomy group had high specific activities of hyaluronidase. The results indicated that hyaluronidase was not only from the testis but also from the male accessory sexual glands. Oligospermic group (Group III) showed the lowest total activity of hyaluronidase among them. The specific activities of ${\beta}$ -N-acetylglucosaminidase was high in oligospermic group (Group III) and low in vasectomy group (Group V). These results were contradictory with the pattern of hyaluronidase activities. This indicated that the spermatozoa which were stayed in epididymis would increase the activity of this enzyme. The specific activity of ${\beta}$ - glucuronidase was low in oligospermic and vasectomy groups. Group VI including testis tumor had remarkably high arylsulfatase activity. Arylsulfatase, a typical lysosomal enzyme, has been known to be released unusually large amounts from certain tumor cells. Arylsulfatase was also released with high activities from azoospermic and vascetomy group. This result indicated that this enzyme was also released from the sources other than testis. Acrosin, a proteolytic enzyme locating in the sperm acrosome, was not found throughout all the samples of seminal plasma. The activities of azocoll proteinase, a non-specific neutral proteinase was nearly identical in all the groups. This enzyme must have been released from the sources other than testis.
Background: Human seminal plasma (HSP)-induced hypersensitivity is one of the serious complications with sexual intercourse. The clinical manifestations of HSP-induced hypersensitivity may be related to the release of vasoactive mediators from mast cell induced by HSP. It has recently been reported that HSP modulates immune systems and induces mast cell degranulation and histamine release from rat peritoneal mast cells (RPMC). Ketotifen and disodium cromoglycate (DSCG), anti-asthmatic and anti-allergic drugs, have a role of mast cell stabilization and inhibit mast cell-induced leukocyte rolling and adhesion. But the inhibitory agents of HSP-induced mast cell activation are unknown. This study was performed to investigate the effects of DSCG and ketotifen on the HSP-induced mast cell activation. Methods: For this, influences of DSCG and ketotifen on the human seminal plasma-induced degranulation, histamine release and morphological changes of RPMC were observed. Results: The mast cell degranulation and histamine release of RPMC by HSP were induced in a dose-dependent fashion. The HSP-induced cytomorphological changes such as swelling, intracellular vacoules, and interrupted cell boundary were significantly inhibited by pretreatment with DSCG or ketotifen. DSCG and Ketotifen inhibited the HSP-induced degranulation and histamine release from RPMC. Conclusion: From the above results, it is suggested that DSCG and ketotifen have a inhibitory effect of the HSP-induced mast cell activation. DSCG and ketotifen may be used for treatment of HSP-induced hypersensitivity.
Lee, Hyo Serk;Park, Yong-Seog;Lee, Joong Shik;Seo, Ju Tae
Clinical and Experimental Reproductive Medicine
/
제43권2호
/
pp.97-101
/
2016
Objective: Growth hormone and its mediator, insulin-like growth factor-1 (IGF-1), have been suggested to exert gonadotropic actions in both humans and animals. The present study was conducted to assess the relationship between serum IGF-1 concentration, seminal plasma concentration, and sperm parameter abnormalities. Methods: A total of 79 men were enrolled in this study from December 2011 to July 2012 and were prospectively analyzed. Patient parameters analyzed included age, body mass index, smoking status, urological history, and fertility history. Patients were divided into four groups based on their semen parameters: normal (A, n=31), abnormal sperm motility (B, n=12), abnormal sperm morphology (C, n=20), and two or more abnormal parameters (D, n=16). Patient seminal plasma and serum IGF-1 concentrations were determined. Results: Patient baseline characteristics were not significantly different between any of the groups. The serum IGF-1 levels in groups B, C, and D were significantly lower than the levels in group A; however, the seminal plasma IGF-1 levels were not significantly different between any of the groups. Conclusion: Men with abnormal sperm parameters had significantly lower levels of serum IGF-1 compared with men with normal sperm parameters. Seminal plasma IGF-1 levels, however, did not differ significantly between the groups investigated here. Further investigations will be required to determine the exact mechanisms by which growth hormone and IGF-1 affect sperm quality.
Objective: In vitro fertilization (IVF) is a well-known method for the treatment of infertility. The present study aimed to compare the differences between infertile women with successful and unsuccessful IVF outcomes regarding the expression of T helper (Th) cell transcription factors and a group of related cytokines before and after exposure to their husbands' seminal plasma. Methods: This study was performed on 19 couples with unexplained infertility undergoing IVF treatment. Among the studied group, nine and 10 couples had successful and unsuccessful IVF outcomes, respectively. This study was carried out using real-time polymerase chain reaction. Results: Before seminal plasma exposure, the expression levels of T-bet (p< 0.007), $interferon-{\gamma}$ (p= 0.013), and tumor necrosis factor $(TNF)-{\alpha}$ (p= 0.017) were higher in the infertile women with IVF failure than in those with successful IVF outcomes, while those of GATA3 (p< 0.001), Foxp3 (p= 0.001), and interleukin (IL)-35 (p< 0.003) were lower. After seminal exposure, the expression of T-bet (p= 0.02), Rorc (p< 0.001), $TNF-{\alpha}$ (p= 0.001), Foxp3 (p= 0.02), and $interferon-{\gamma}$ (p= 0.001) increased in the unsuccessful IVF group, while the expression of Foxp3 (p= 0.02), Rorc (p< 0.001), IL-23 (p= 0.04), IL-17 (p= 0.02), IL-6 (p< 0.001), transforming growth $factor-{\beta}$ (p= 0.01), and IL-35 (p< 0.001) increased in the successful IVF group. Conclusion: In summary, IVF failure was associated with imbalanced Th1/Th2/Th17/Treg responses. Moreover, our results show that seminal plasma might have a positive effect on IVF outcomes via changes in peripheral blood T cell subsets.
Despite a considerable amount of investigation there continues to be disagreement concerning the proteins present in human seminal plasma. Recently their identification has assumed a greater importance following evidence that infertility in men and women may have an immunological cause (Katsh, 1959; Quinlivan, 1969). Seminal plasma is composed of fluids secreted by the prostate, seminal vesicles, ampullae, ducti deferentes, bulbourethral (Cowper's) glands, urethral(Littre's) glands and the epididymes. Prostatic juice, one of the major components of seminal plasma, has an important role in secretion of acid phosphatase and prostaglandin. A few studies have been reported of human prostatic juice, since, in human subjects, there were some problems in studying prostatic juice due to quite small amount of secretion and possibility of contamination with fluids from the seminal vesicles and ejaculatory ducts. The purpose of the present study was to determine the basic components of proteins in human prostatic juice. Prostatic juice was obtained from normal healthy man of $20{\sim}30\;year-old$ by massage of the prostate, and protein components were separated by means of disc electrophoresis. The results are summarized as follows; 1) Total numbers of protein fractions of normal human serum and prostatic juice are $14{\sim}18$ bands and $9{\sim}12$ bands, respectively. Prostatic juice produces two deeply staining bands which appear similar to those formed by $beta-_1$ globulin and albumin. 2) $Alpha-_1$ globulin area in the fractions of prostatic juice shows 4 bands and one more band is found than that of serum. On the other hand, the fractions of immunoglobulin and $alpha-_2$ globulin areas are eight in serum and it has three bands more than that of prostatic juice. 3) $Alpha-_1$ globulin area in the prostatic juice is more deeply stained than that of serum. In contrast with $alpha-_1$ globulin area, immunoglobulin and $alpha-_2$ globulin areas in the prostatic juice show weaker staining than serum.
The efficiency of artificial insemination (AI) for horses remains unsatisfactory. It is mainly because each process of AI causes a detrimental effect on semen quality. To sustain quality of semen properly, several factors including libido of stallions and sperm damage during sperm processing and preservation should be considered. Stallions with decent libido produce a high ratio of sperm to seminal plasma in their ejaculates, which is the ideal semen composition for maintaining sperm quality. Thus, to maximize the fertility rate upon AI, stallions should be appropriately managed to enhance their libido. Seminal plasma should have a positive effect on horse fertility in the case of natural breeding, whereas the effects of seminal plasma on both sperm viability and quality in the context of AI remain controversial. Centrifugation of semen is performed during semen processing to remove seminal plasma and to isolate fine quality sperm from semen. However, the centrifugation process can also result in sperm loss and damage. To solve this problem, several different centrifugation techniques such as Cushion Fluid along with dual and single Androcoll-E$^{TM}$ were developed to minimize loss of sperm and to damage at the bottom of the pellet. Most recently, a new technique without centrifugation was developed with the purpose of separating sperm from semen. AI techniques have been advanced to deliver sperm to optimal region of female reproductive tract at perfect timing. Recombinant equine luteinizing hormone (reLH) and low dose insemination techniques have been developed to maximize both fertility rate and the efficiency of AI. Horse breeders should consider that the entire AI procedure should be optimized for each stallion due to variation in individual horses for a uniformed AI protocol.
The total proteins were estimated in both deoxycholate (DOC)-extract of sperm membrane and seminal plasma of chilled as well as frozen semen obtained from five Murrah buffalo bulls. Proteins were further characterized by polyacrylamide gel electrophoresis (PAGE) in three bulls. The protein content of sperm membrane extract (SME) and that of seminal plasma (SP) decreased gradually with increase in freezing period from 6 to 24 mo when compared with the values observed in freshly chilled semen in all bulls. The total decrease in protein content of SME and SP varied from 30-40% and 28-59% respectively during 6-24 mo of freezing. The number of glycoproteins/proteins (GP/P) in SME varied from 4-8 in freshly-chilled semen of all bulls and reduced to 2-4 after 24 mo of freezing. In SP, the number of proteins varied from 6-10 in freshly chilled semen of all bulls and reduced to 3-8 after 24 mo of freezing. Some of the proteins in SME and SP disappeared, others got altered and appeared with change in molecular weight after different freezing times. These studies reveal that alterations in the sperm membrane proteins may be responsible for damage to their membrane during freezing and thus lowering their fertilizability.
Objective: Oxidative stress plays a key role in the pathogenesis of male infertility. But, the adverse effects of oxidative biomarkers on sperm quality remain unclear. This study aimed to investigate the levels of nitric oxide (NO), 8-hydroxydesoxyguanosine (8-OHdG), and total antioxidant capacity (TAC) oxidative biomarkers in seminal plasma and their relationship with sperm parameters. Methods: A total of 77 volunteers participated in the study, including fertile (n = 40) and infertile men (n = 37). NO, 8-OHdG, and TAC levels were measured using the ferric reducing ability of plasma, Griess reagent method and an enzyme-linked immunosorbent assay kit, respectively. Results: The mean values of sperm parameters in the infertile group were significantly lower than those in the fertile group (p< 0.001). The mean 8-OHdG in the seminal plasma of infertile men was significantly higher (p= 0.013) than those of controls, while the mean TAC was significantly lower (p= 0.046). There was no significant difference in NO level between the two groups. The elevated seminal 8-OHdG levels were negatively correlated with semen volume, total sperm counts and morphology (p< 0.001, p= 0.001 and p= 0.052, respectively). NO levels were negatively correlated with semen volume, total sperm counts and morphology (p= 0.014, p= 0.020 and p= 0.060, respectively). Positive correlations between TAC and both sperm count and morphology (p= 0.043 and p= 0.025, respectively) were also found. Conclusion: These results suggested that increased levels of NO and 8-OHdG in seminal plasma could have a negative effect on sperm function by inducing damage to the sperm DNA hence their fertility potentials. Therefore, these biomarkers can be useful in the diagnosis and treatment of male infertility.
Heparin binding proteins (HBPs) are produced by accessory glands. These are secreted into the seminal fluid, bind to the spermatozoa at the time of ejaculation, favour capacitation, acrosome reaction, and alter the immune system response toward the sperm. The present study was conducted with an objective to assess the effect of purified seminal plasma-HBPs (SP-HBPs) on cross bred cattle bull sperm attributes during two phases of cryopreservation: Pre freezing and freezing-thawing. SP-HBPs were purified from pooled seminal plasma by heparin affinity chromatography. Three doses of SP-HBPs i.e. 10, 20, $40{\mu}g/mLs$ semen were standardized to find out the optimum dose and $20{\mu}g/mLs$ was found to be an optimum dose. Semen as such and treated with SP-HBPs was diluted with sodium citrate-egg yolk diluter and cryopreserved as per the standard protocol. Sperm parameters i.e. motility, viability, Hypo-osmotic swelling test (HOST), acrosome damage, in vitro capacitation and lipid peroxidation were evaluated in SP-HBP treated and untreated (control) semen at both phases of cryopreservation. A considerable variation in percent sperm motility, viability, membrane integrity (HOST), acrosome damage, acrosome reaction and lipid peroxidation was observed at both phases among the bulls irrespective of the treatment. Incubation of neat semen with $20{\mu}g/mL$ SP-HBP before processing for cryopreservation enhanced the average motility, viability, membrane integrity by 7.2%, 1.5%, 7.9%, and 5.6%, 6.6%, 7.4% in pre-frozen and frozen-thawed semen in comparison to control. There was also an average increase of 4.1%/3.9% in in vitro capacitation and acrosome reaction in SP-HBPs-treated frozen-thawed semen as compared to control. However, binding of SP-HBPs to the sperm declined acrosome damage and lipid peroxidation by 1.3%/4.1% and 22.1/$32.7{\mu}M$/$10^9$ spermatozoa in SP-HBP treated pre-frozen/frozen-thawed semen as compared to control, respectively. Significant (p<0.05) effects were observed only in motility, HOST and in vitro acrosome reaction. It can be concluded that treatment of neat semen with SP-HBPs before cryopreservation minimized the cryoinjury by decreasing the generation of reactive oxygen species.
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