scholarly journals Papain and its inhibitor E-64 reduce camelid semen viscosity without impairing sperm function and improve post-thaw motility rates

2017 ◽  
Vol 29 (6) ◽  
pp. 1107 ◽  
Author(s):  
C. M. Kershaw ◽  
G. Evans ◽  
R. Rodney ◽  
W. M. C. Maxwell
Keyword(s):  
Sperm Motility ◽  
Exposure Time ◽  
Seminal Plasma ◽  
Assisted Reproductive Technologies ◽  
Reproductive Technologies ◽  
Plasma Viscosity ◽  
Dna Integrity ◽  
Sperm Function ◽  
Viscous Component ◽  
Acrosome Integrity

In camelids, the development of assisted reproductive technologies is impaired by the viscous nature of the semen. The protease papain has shown promise in reducing viscosity, although its effect on sperm integrity is unknown. The present study determined the optimal papain concentration and exposure time to reduce seminal plasma viscosity and investigated the effect of papain and its inhibitor E-64 on sperm function and cryopreservation in alpacas. Papain (0.1 mg mL–1, 20 min, 37°C) eliminated alpaca semen viscosity while maintaining sperm motility, viability, acrosome integrity and DNA integrity. Furthermore E-64 (10 µM at 37°C for 5 min after 20 min papain) inhibited the papain without impairing sperm function. Cryopreserved, papain-treated alpaca spermatozoa exhibited higher total motility rates after chilling and 0 and 1 h after thawing compared with control (untreated) samples. Papain treatment, followed by inhibition of papain with E-64, is effective in reducing alpaca seminal plasma viscosity without impairing sperm integrity and improves post-thaw motility rates of cryopreserved alpaca spermatozoa. The use of the combination of papain and E-64 to eliminate the viscous component of camelid semen may aid the development of assisted reproductive technologies in camelids.

scholarly journals From One Ejaculate to Another: Transference of Sperm Traits via Seminal Plasma Supplementation in the Ram

Biology ◽  
2020 ◽  
Vol 9 (2) ◽  
pp. 33
Author(s):  
Christine Green ◽  
Jessica P. Rickard ◽  
Simon P. de Graaf ◽  
Angela J. Crean
Keyword(s):  
Sperm Motility ◽  
Seminal Plasma ◽  
Perceived Risk ◽  
Positive Relationship ◽  
Assisted Reproductive Technologies ◽  
Sperm Quality ◽  
Reproductive Technologies ◽  
Before And After ◽  
The Difference ◽  
Sperm Traits

Males can adjust sperm motility instantaneously in response to the perceived risk of sperm competition. The speed of this response suggests that sperm motility is regulated by changes in seminal plasma rather than changes in the sperm cells themselves. Hence, here we test whether inter-ejaculate variation in seminal plasma can be used to alter sperm quality prior to use in assisted reproductive technologies. We supplemented fresh ejaculates of Merino rams with seminal plasma collected from previous ‘donor’ ejaculates to test whether changes in sperm kinetics were related to the relative quality of donor to focal ejaculates. We found a positive relationship between the change in sperm traits before and after supplementation, and the difference in sperm traits between the donor and focal ejaculate. Hence, sperm motility can be either increased or decreased through the addition of seminal plasma from a superior or inferior ejaculate, respectively. This positive relationship held true even when seminal plasma was added from a previous ejaculate of the same ram, although the slope of the relationship depended on the identity of both the donor and receiver ram. These findings indicate that seminal plasma plays a key role in the control and regulation of sperm kinetics, and that sperm kinetic traits can be transferred from one ejaculate to another through seminal plasma supplementation.

scholarly journals Structure-based redesigning of pentoxifylline analogs against selective phosphodiesterases to modulate sperm functional competence for assisted reproductive technologies

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Mutyala Satish ◽  
Sandhya Kumari ◽  
Waghela Deeksha ◽  
Suman Abhishek ◽  
Kulhar Nitin ◽  
...  
Keyword(s):  
Sperm Motility ◽  
In Silico ◽  
Assisted Reproductive Technologies ◽  
Ex Vivo ◽  
Binding Free Energy ◽  
Reproductive Technologies ◽  
Early Embryo ◽  
Dna Integrity ◽  
Binding Studies

AbstractPhosphodiesterase (PDE) inhibitors, such as pentoxifylline (PTX), are used as pharmacological agents to enhance sperm motility in assisted reproductive technology (ART), mainly to aid the selection of viable sperm in asthenozoospermic ejaculates and testicular spermatozoa, prior to intracytoplasmic sperm injection (ICSI). However, PTX is reported to induce premature acrosome reaction (AR) and, exert toxic effects on oocyte function and early embryo development. Additionally, in vitro binding studies as well as computational binding free energy (ΔGbind) suggest that PTX exhibits weak binding to sperm PDEs, indicating room for improvement. Aiming to reduce the adverse effects and to enhance the sperm motility, we designed and studied PTX analogues. Using structure-guided in silico approach and by considering the physico-chemical properties of the binding pocket of the PDEs, designed analogues of PTX. In silico assessments indicated that PTX analogues bind more tightly to PDEs and form stable complexes. Particularly, ex vivo evaluation of sperm treated with one of the PTX analogues (PTXm-1), showed comparable beneficial effect at much lower concentration—slower AR, higher DNA integrity and extended longevity of  spermatozoa and  superior embryo quality. PTXm-1 is proposed to be a better pharmacological agent for ART than PTX for sperm function enhancement.

scholarly journals A Comparison Between the Effects of Global Sperm Washing® and FertiCult Flushing TM Media on Certain Sperm Function Parameters of Asthenozoospermic Men

2017 ◽  
Vol 11 (2) ◽  
pp. 37-41
Author(s):  
Saad S. Al-Dujaily ◽  
Khalid Al-Azzawi ◽  
Zena Hussein ◽  
Ban Al-Anii
Keyword(s):  
Sperm Motility ◽  
Assisted Reproductive Technologies ◽  
Semen Analysis ◽  
Reproductive Technologies ◽  
World Health ◽  
Sperm Function ◽  
Sperm Activation ◽  
Optimum Result ◽  
Sperm Washing

The World Health Organization (WHO) and many studies considered the infertility as a disease and so many couples complaining from unsuccessful assisted reproductive technologies procedures to overcome their problem. One of the reasons of this dilemma is the sperm preparation method when no optimum result obtained even by using any of media found globally. However Global sperm washing®, and FertiCult flushingTM media were proved their capability to obtain good results of certain sperm function parameters. Nevertheless, the studies that compare between these media were rare. Therefore, this study aimed to compare between Global sperm washing medium®, FertiCult flushing TM media that used for sperm washing before using the partner sperm for ART procedure. After detecting asthenozoospermia in sixty semen samples, they were divided into two groups according to medium used for sperm activation in vitro Global sperm washing medium ® (n=31) and FertiCult flushing mediumTM (n=29) groups.The semen analysis was done after 3-5 days of abstinence as recommended by the manual of WHO (1999). Certain sperm function parameters were recorded. Semen fluid samples were treated with sperm activation media (Global sperm washing medium and FertiCult flushing medium TM) by using direct swim-up technique for in vitro sperm activation test. A significant (P<0.05) improvement was noticed between the two media regarding active sperm motility grades A and  B when using FertiCult flushing mediumTM compared to Global sperm washing medium®. Whereas no significant (P>0.05) differences were detected between the two media regarding sperm motility grades C and D. There was no significant (P>0.05) differences in morphologically normal sperm  following in vitro activation by using the two media. It is concluded that FertiCult flushing mediumTM was better than Global sperm washing medium®  in improving active sperm motility of asthenozoospermic men which can be utilized in future for successful of assisted reproduction.

Glycosaminoglycans in the accessory sex glands, testes and seminal plasma of alpaca and ram

2012 ◽  
Vol 24 (2) ◽  
pp. 362 ◽  
Author(s):  
Claire M. Kershaw-Young ◽  
G. Evans ◽  
W. M. C. Maxwell
Keyword(s):  
Seminal Plasma ◽  
Assisted Reproductive Technologies ◽  
Prostate Gland ◽  
Keratan Sulfate ◽  
Reproductive Technologies ◽  
Plasma Viscosity ◽  
Accessory Sex Glands ◽  
The Relationship

The viscous nature of alpaca semen limits its use in cryopreservation and other assisted reproductive technologies. The cause and source of this viscosity is unknown although it has been postulated, but never proven, that glycosaminoglycans (GAGs) secreted by the bulbourethral gland are responsible. The present study investigated the concentration and composition of GAGs in alpaca seminal plasma, testes, bulbourethral gland and prostate gland and compared them to those in the ram to determine the relationship between seminal plasma GAGs and viscosity and to identify the source of seminal plasma GAGs. Alpaca seminal plasma contained more GAGs than ram (P < 0.001) and the predominant GAG, keratan sulfate, was correlated with viscosity (P = 0.05, R2 = 0.2635). The alpaca bulbourethral gland contained most GAGs compared with prostate or testis (P < 0.001). In the ram, the prostate contained most GAGs. These findings suggest that GAGs, particularly keratan sulfate, may be the cause of seminal plasma viscosity in alpacas, and that the seminal plasma GAGs originate from the bulbourethral gland.

P–050 The effectiveness of the platelet-rich plasma treatment of men with severe oligoasthenoteratozoospermia

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
O Somova ◽  
H Ivanova ◽  
N Sotnyk ◽  
K Kovalenko ◽  
I Feskova
Keyword(s):  
Male Infertility ◽  
Sperm Motility ◽  
Testosterone Level ◽  
Assisted Reproductive Technologies ◽  
Platelet Rich Plasma ◽  
Sperm Concentration ◽  
Free Testosterone ◽  
Reproductive Technologies ◽  
Positive Effects ◽  
Free Testosterone Level

Abstract Study question To evaluate the effect of platelet-rich plasma (PRP) testicular injections on spermogram parameters of men with severe oligoasthenoteratozoospermia (OAT). Summary answer The PRP testicular injections have beneficial effects on spermatogenesis and enhance sperm concentration and motility in infertile men with OAT. What is known already The use of PRP therapy in assisted reproductive technologies is debatable. Despite the recent evidence of its positive effects in promoting endometrial and follicular growth, data from clinical studies are limited. There are only a few papers on the effectiveness of PRP therapy in the treatment of male infertility and sexual dysfunction. In more detail, the influence of PRP on spermatogenesis was carried out only on experimental animals. Although the mechanisms of its action have not yet been clarified, it is assumed that PRP, containing many biologically active molecules, realizes its effect through the tissue regeneration and cell proliferation. Study design, size, duration This prospective study included 68 men (34.6±5.2) years old with severe OAT (≤4 million/ml, motility ≤30%, normal sperm morphology ≤1%) receiving hormonal and antioxidant (AO) therapy during 6 months before in vitro fertilization cycles. 33 of them were injected once with autologous PRP (0.5 ml in each testicle). Spermogram and testosterone level were analyzed before the treatment and in 3, 4 and 6 months after it. Participants/materials, setting, methods: Sperm concentration, motility and morphology in ejaculate of 33 men of PRP group were compared with those in the group of 35 men without PRP within 6 months of starting the treatment. Total and free testosterone level were measured in blood serum. PRP was prepared by centrifuging the patient’s own blood in the anticoagulant-containing tubes. The final concentration of platelets in the obtained sample was 950.000 – 1.250 000 cells in 1 ml. Main results and the role of chance 4 months after the PRP injection, sperm concentration and motility increased in 18 of 33 men of the PRP group compared with the baseline (before the treatment) – 4.2 (1.0; 6.9) vs 1.4 (0.1; 3.4) mln/ml (p &lt; 0.05) and 36.7 (30.6; 45.8) vs 17.7 (6.7; 28.2)% respectively (p &lt; 0.05).The maximum increase in sperm motility (but not in sperm concentration!) was observed in 24 men in 6 months – 49.6 (39.6; 56.4)% (p &lt; 0.05). Percent of morphologically normal spermatozoa in ejaculate slightly increased only in 12 men in that time period from 0–1% to 1–2%. The total testosterone level was 2.4 times higher than the baseline (31.6±7.2 vs 13.2±4.3 nmol/l, p &lt; 0.05), the free testosterone level was 1.8 times higher (14.5±3.5 vs 7.9±3.0 pgl/ml, p &lt; 0.05). Unlike the PRP group, in the group of men without PRP treatment, the sperm parameters did not changed compared with the baseline in 4 months after the starting hormonal and AO treatment. A significant increase of sperin concentration was observed only in 17 of 35 patients in 6 months. Sperm motility and percent of morphologically normal spermatozoa after the treatment did not differ from the baseline. Changes in the testosterone levels were similar to changes in PRP group. Limitations, reasons for caution Only young and middle-aged men were considered in the study. Large randomized controlled studies are required to confirm the PRP therapy efficacy and safety of f various fertility disorders. There are also no standardized protocols for PRP preparation. Wider implications of the findings: PRP therapy may have great potential for the treatment of male infertility and improving spermatogenesis. Optimization of methods of PRP preparation and dosage of testicular injections can enhance reproductive outcomes in assisted reproductive technologies. Trial registration number Not applicable

97 IMPROVED CRYOPRESERVATION OF DOMESTIC CAT SPERMATOZOA IN A SOY LECITHIN-BASED EXTENDER

2011 ◽  
Vol 23 (1) ◽  
pp. 153 ◽  
Author(s):  
M. M. Vick ◽  
H. L. Bateman ◽  
W. F. Swanson
Keyword(s):  
Sperm Motility ◽  
Room Temperature ◽  
Egg Yolk ◽  
Tris Buffer ◽  
Domestic Cat ◽  
Dna Integrity ◽  
Freezing And Thawing ◽  
Soy Lecithin ◽  
Acridine Orange Staining ◽  
Acrosome Integrity

Development of a chemically defined, plant-based cryopreservation media would reduce extender variability and the potential for transmission of zoonotic pathogens compared with traditional egg-yolk-based extenders. The objective of this study was to compare effects of egg yolk- and soy lecithin-based cryopreservation media and the temperature of glycerol addition on sperm parameters following freezing and thawing of domestic cat spermatozoa. Fresh semen was collected by manual stimulation on 3 separate occasions from 4 adult male cats. Each ejaculate was washed to remove seminal plasma, divided into 4 equal aliquots, and extended at room temperature in one of the following treatments: 1) TEST-egg yolk (Irvine Scientific Inc., Santa Ana, CA, USA) medium with 4% glycerol (EYG); 2) TEST-egg yolk, with 4% glycerol added after cooling to 5°C (EY); 3) TES-Tris buffer with soy-lecithin (1%) and 4% glycerol (SLG); and 4) TES-Tris buffer with 1% soy-lecithin, and 4% glycerol added after cooling to 5°C (SL). Sperm progressive motility (%) and rate of progressive movement (scale of 0–5) were evaluated at 0, 1, 3, 6, and 24 h post-thaw. Sperm capacitation (chlortetracycline staining), acrosome integrity (FITC-PNA staining), and DNA integrity (acridine orange staining) were assessed at 15 min post-thaw. Data were exponentially transformed to achieve normal distribution and then subjected to GLM analysis to determine effects of media and temperature of glycerol addition on sperm traits. At 0 and 1 h post-thaw, acrosome integrity, DNA integrity and % sperm motility did not differ (P > 0.05) among treatments. However, % sperm motility was greater in the soy-based media compared to egg yolk-based media at 3, 6, and 24 h post-thaw (Table 1; P < 0.05). A higher percentage of uncapacitated spermatozoa were present in soy-based compared to egg-yolk based cryopreservation media (63.9 ± 9.3 v. 51.2 ± 11.5, respectively; P < 0.05), regardless of temperature of glycerol addition. Finally, addition of glycerol at 5°C resulted in higher % sperm motility compared to room temperature at 6 and 24 h post-thaw in both medium types (Table 1; P < 0.05). Our results suggest that use of a chemically defined, soy-based medium improves long-term motility and capacitation status of frozen–thawed domestic cat spermatozoa compared with cryopreservation in a traditional egg yolk-based extender. Table 1.Motile spermatazoa and motility score at 3, 6, and 24 h

163 EVALUATION OF VIABILITY OF BULL SEMEN COLLECTED BY ELECTRO-EJACULATION USING COMMERCIAL SEMEN EXTENDER AND 2 CULTURE MEDIA AT CONTROLLED ROOM TEMPERATURE

2017 ◽  
Vol 29 (1) ◽  
pp. 190
Author(s):  
A. M. Raseona ◽  
O. A. Ajao ◽  
L. D. Nethengwe ◽  
L. R. Madzhie ◽  
T. L. Nedambale ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Assisted Reproductive Technologies ◽  
Culture Media ◽  
Reproductive Technologies ◽  
Sperm Viability ◽  
Progressive Motility ◽  
Bull Semen ◽  
Significant Difference ◽  
Semen Extender ◽  
Motility Rate

Preservation of semen is an important process to ensure that semen quality is sufficient for assisted reproductive technologies. The aim of this study was to evaluate the viability of bull semen collected by electro-ejaculation using commercial semen extender and 2 modified culture media stored at controlled RT (24°C) for 72 h. Two Nguni bulls were used for semen collection; after collection, the semen was evaluated macroscopically for volume, pH, and colour, and microscopically for sperm motility, viability, and morphology. Uncontaminated semen samples with progressive motility >70% and morphological defects <20% were pooled after collection before being aliquoted into 3 extenders, namely Triladyl, modified Ham’s F10, and TCM-199 culture media, at a dilution ratio of 1:4 and then stored at controlled RT (24°C). Sperm motility rate was analysed using the computer-aided sperm analyser after 0, 24, 48, and 72 h of storage. Sperm morphology and viability was performed after staining the sperm cells with spermac and nigrosine-eosin stain, respectively. The study was replicated 4 times and data were analysed using ANOVA. Triladyl had a higher sperm viability rate (41.3%) and total motility rate (96.3%) for 72 h (P < 0.01) compared with the 2 modified culture media, Ham’s F10 (26.5 and 86.8%) and TCM-199 (25.0 and 86.7%), respectively. However, Ham’s F10 had higher progressive motility rate (37.8%) as compared with the other extenders, TCM-199 (31.7%) and Triladyl (23.4). There was no significant difference (P > 0.05), in viability rate between Ham’s F10 (26.5%) and TCM-199 (25.0%). No significant difference (P > 0.05) in straight line velocity was observed for the three extenders. Furthermore, no significant difference was observed in total sperm abnormalities, except for reacted acrosomes and absent tails (P > 0.05), between the 2 Nguni bulls. Nguni semen can be preserved in Triladyl or modified Ham’s F10 and TCM-199 culture media stored at 24°C and stay viable for 72 h. Triladyl proved to be the best suitable extender of the 3 extenders, showing higher sperm viability and total motility rate as compared with Ham’s F10 and TCM-199 modified culture media.

108 Synchronization and artificial insemination of South African communal cattle

2021 ◽  
Vol 33 (2) ◽  
pp. 161
Author(s):  
T. L. Magopa ◽  
M. L. Mphaphathi ◽  
T. Mulaudzi ◽  
F. V. Ramukhithi ◽  
M. M. Tshabalala ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Assisted Reproductive Technologies ◽  
Bos Taurus ◽  
Body Condition Score ◽  
Bos Indicus ◽  
Prostaglandin F2α ◽  
Reproductive Technologies ◽  
Computer Assisted ◽  
Bos Taurus Indicus ◽  
Oestradiol Benzoate

The present study aimed to evaluate an oestrous synchronization protocol and oestrus response before AI in cows from an organised communal production system. A total of 74 cows Bos indicus (Brahman) and Bos taurus/indicus hybrid (Nguni) type from different communal farmers were screened based on: age (3 to 8 years), body condition score of ≥3 (1–5 scale), not pregnant (excluding heifers), 90 days postpartum, number of parities, good mothering ability, and negative for contagious abortion. Selected cows were subjected to a 9-day OvSynch+CIDR protocol. In brief, the protocol included the insertion of controlled intravaginal drug release (CIDR®, Pfizer Laboratories) devices in the reproductive tract (vagina) containing 1.9g of progesterone in each cow on Day 0 with first oestradiol benzoate (Oestradiol benzoate®, VTech) 2-mL intramuscular (IM) injection. Pregnant mare serum gonadotrophin (PMSG; Chronogest®, Intervet International B.V.) 2.5-mL (IM) injection on Day 5. Removal of CIDR and (IM) injection of prostaglandin F2α (PGF2α; Estrumate®, Intervet South Africa (Pty.) Ltd.) 2mL on Day 8. Following CIDR removal, a heat mount detector (Kamar®) was applied on the individual cow’s tail head for oestrus observation (signs of heat) with second oestradiol benzoate 1-mL (IM) injection on Day 9. AI was performed 36h following withdrawal of the CIDR. Frozen/thawed semen from Bonsmara bulls (n=2) of known and proven fertility was used for AI. The GameteTek Cryo-Mobile laboratory was used during thawing of semen straws. and sperm motility and morphological traits were immediately evaluated by computer-assisted sperm analysis (Sperm Class Analyzer®) before each individual cow was inseminated. Pregnancy diagnosis was performed 90 days after timed AI by ultrasound and transrectal palpation. Data were analysed using the logistic regression procedure of SAS (SAS Institute Inc.), with synchronisation response and conception being treated as binary response variables. All cows were synchronized successfully and an oestrus response rate of 100% was recorded. Pregnancy rates were similar (37.8% vs. 38.9%) for both Bos indicus and Bos taurus/indicus hybrid cattle. Bulls total sperm motility of ≥75% were recorded, following thawing of semen straws. Thus, there was no effect of bull on pregnancy. In conclusion, acceptable oestrus synchronization response was achieved in this communal setup. Superior genetic materials can be successfully introduced through assisted reproductive technologies in organised communal production systems.

229 From head to tail: A red wolf sperm project

2020 ◽  
Vol 32 (2) ◽  
pp. 242
Author(s):  
S. Kamen ◽  
J. Nagashima ◽  
N. Songsasen ◽  
M. Ferraz
Keyword(s):  
Sperm Motility ◽  
Proteomic Analysis ◽  
Assisted Reproductive Technologies ◽  
Culture Media ◽  
Egg Yolk ◽  
Reproductive Technologies ◽  
Canis Rufus ◽  
Red Wolf ◽  
Paired Samples ◽  
Sperm Survival

Development of assisted reproductive technologies for the critically endangered red wolf (Canis rufus) is crucial to the maintenance of genetic diversity to support species recovery. Towards this goal, a cryopreservation protocol has previously been developed for red wolf sperm; however, the ability of the gametes to undergo capacitation has not been assessed in this species. Previously, we have shown that oviductal extracellular vesicles (oEVs) improve cat sperm motility and fertilizing ability. The objectives were to (1) compare the effects of culture media on motility and acrosomal integrity of fresh sperm, and select the best medium that can be used in a capacitation protocol; (2) identify potential biomarkers for sperm cryo-tolerance; and (3) determine the influence of canine oEVs on sperm survival and motility post-thaw. In Study 1, sperm were collected by electro-ejaculation from adult red wolves (n=8) and immediately cryopreserved in TRIS-egg yolk buffer with 8% glycerol or incubated for 18h in 5% CO2 and 38.5°C in one of the following media: canine capacitation medium (CCM), FERT-TALP (FERT), NCSU, synthetic oviductal fluid (SOF) and TRIS. At 0, 1, 2, 3, 4, and 18h, sperm were evaluated for total motility and acrosome integrity (FITC-PNA). In Study 2, sperm with high (&gt;80%, HM, n=2 wolves) and low (&lt;15%, LM, n=2 wolves) motility post-incubation at 4°C in the cryopreservation medium for 18h were subjected to proteomic analysis. In Study 3, oviducts were collected from domestic dogs (1-9 years, n=12) after elective spaying, and oEVs from various stages of the oestrous cycle [early follicular (EF), late follicular (LF), early luteal (EL), and late luteal (LL)] were isolated using the Total Exosome Isolation kit (Invitrogen). Frozen-thawed red wolf sperm (n=4 males) were incubated with 30×106 oEVs in non-capacitating CCM, and assessed as in study 1 at 0, 0.5, 1, 2, 3, 4, 6, 8, and 10h. Data were analysed using a paired samples t-test with 95% CI (Prism8, GraphPad Inc.). Sperm incubated in CCM and NCSU had higher motility than those in FERT, SOF, and TRIS after 2h of incubation and onward (2 h: 65±6, 68±6, 42±10, 57±8, and 43±5; 3 h: 60±9, 63±8, 36±11, 46±9, and 34±6; 4 h: 60±9, 60±10, 30±10, 43±8, and 20±5; 18 h: 12±7, 15±7, 9±5, 3±2, and 0, respectively; P&lt;0.05). After 1h of incubation, samples incubated in CCM, NCSU, and SOF had a higher number of sperm with intact acrosomal membranes (P&gt;0.05) than other treatments. A total of 179 proteins were identified, of which 129, including those regulating energy metabolism and mitochondrial mediated apoptosis, were differentially expressed between HM and LM. Preliminary data from Study 3 suggested that thawing and incubating sperm in the presence of LF, EL, and LL oEVs improved sperm motility. In conclusion, CCM and NCSU sustained sperm survival after invitro incubation and could be candidates for invitro fertilization studies in the red wolf. Data generated from sperm proteomic analysis provided insights into cellular pathways regulating sperm cryo-sensitivity. Finally, we demonstrated the potential of oEVs in improving wolf sperm survival post-thawing.

Features of seminal plasma cytokine profile and an occurrence of pregnancy when carrying out a program of assisted reproductive technologies

GYNECOLOGY ◽  
2019 ◽  
Vol 21 (4) ◽  
pp. 9-13
Author(s):  
Tatiana V Dontsova ◽  
Alina A Babayan ◽  
Marina A Nikolaeva ◽  
Alla S Arefieva ◽  
Elena A Kalinina ◽  
...  
Keyword(s):  
Sexual Intercourse ◽  
Seminal Plasma ◽  
Transforming Growth Factor ◽  
Assisted Reproductive Technologies ◽  
Reproductive Technologies ◽  
Female Reproductive Tract ◽  
Cytokine Profile ◽  
Sexual Life ◽  
Operating Characteristics ◽  
Diagnostic Significance

Aim. To asses an effectiveness of assisted reproductive technologies (ART) program given characteristics of the cytokine profile of seminal plasma (SP) entering the female reproductive tract during sexual intercourse. Outcomes and methods. 33 married couples who applied for a treatment of infertility by means of in vitro fertilization /ICSI (Intracytoplasmic Sperm Injection) were included in the prospective study. Patients were recommended to have sexual intercourse with no restrictions during treatment and to have the last sexual intercourse 3 days before an intended transvaginal puncture (ITP). Testing of cytokines (transforming growth factor (TGF)-b1, interferon (IFN)-g, interleukin (IL)-33, IL-6, IL-8, IL-23, IL-10, tumor necrosis factor (TNF)-a, IL-18, IL- 17A, IFN-a, IL-12, monocyte chemotactic protein-1) levels in samples of partners’ SP obtained on the day having ITP were carried out using multiplex analysis with LegendPlex kits (BioLegend, USA). Results. When comparing a cytokine profile of SP in couples who did not become pregnant (n=25) and couples who become pregnant (n=8) increased IL-18 and reduced IL-10 levels (p=0,017 and p=0, 01 respectively) were revealed in the group which got pregnant. To assess a clinical relevance of cytokine content in SP ROC (Receiver Operating Characteristics) curve was used. It was established that determining of IL-18 concentration in SP has the greatest diagnostic significance (the area under a curve was 0.792±0.107, test sensitivity - 62.5%, test specificity - 95.24% at threshold concentration>210.43 pg/ml). Incidence of pregnancy at IL-18 concentration above threshold levels was 83,3% while at lower concentrations of IL-18 pregnancy occurred only in 13.0% of women. Conclusions. Elevated IL-18 levels and decreased IL-10 levels in SP of female patients’ partners who don’t have restriction of sexual life when treating infertility with ART are favorable factors for a pregnancy to occur.

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