scholarly journals Gelatinases in Boar Seminal Plasma and Their Relation to Semen Indicators

2010 ◽  
Vol 79 (3) ◽  
pp. 491-496 ◽  
Author(s):  
Maja Zakošek Pipan ◽  
Marjan Kosec ◽  
Janko Mrkun ◽  
Petra Zrimšek
Keyword(s):  
Molecular Weight ◽  
Enzyme Activity ◽  
Seminal Plasma ◽  
Animal Species ◽  
Sperm Concentration ◽  
Gelatin Zymography ◽  
Motile Sperm ◽  
Molecular Weights ◽  
Simultaneous Identification ◽  
First Time

Matrix metalloproteinases were detected in reproductive tissues and seminal plasma of various animal species. The aim of this study was to determine for the first time the presence of gelatinases and metalloproteases in boar seminal plasma and to correlate the results with semen indicators. Gelatin zymography was used for simultaneous identification and measurement of gelatinase enzyme activity associated with their molecular weights. Several gelatinase forms were identified in seminal plasma of boars. Those that were stimulated by CaCl2 and inhibited by EDTA and phenanthroline were considered as metalloproteases. Negative correlation between semen indicators (sperm index, sperm concentration and concentration of progressive motile sperm) and the concentrations of metalloprotease at 78 kDa and 66 kDa means that higher values of semen indicators correlate with lower concentrations of these metaloproteases in seminal plasma. Gelatinases with molecular weight of 225, 78 and 66 kDa correlated with higher levels of acrosome damage. Samples with sperm index above 110 M/ml contained gelatinases of significantly lower band intensities at 78 and 66 kDa compared to samples with SI less than 110 M/ml. Bands with 225, 78 and 66 kDa are suggested to belong to a dimer of MMP-9, proMMP-2 and mature MMP-2.

scholarly journals Specific Seminal Plasma Fractions Are Responsible for the Modulation of Sperm–PMN Binding in the Donkey

Animals ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 1388
Author(s):  
Jordi Miró ◽  
Jaime Catalán ◽  
Henar Marín ◽  
Iván Yánez-Ortiz ◽  
Marc Yeste
Keyword(s):  
Molecular Weight ◽  
Sperm Motility ◽  
Seminal Plasma ◽  
Potential Effect ◽  
Polymorphonuclear Neutrophils ◽  
Low Fertility ◽  
Plasma Fractions ◽  
Complex Fluid ◽  
First Time

While artificial insemination (AI) with frozen-thawed sperm results in low fertility rates in donkeys, the addition of seminal plasma, removed during cryopreservation, partially counteracts that reduction. Related to this, an apparent inflammatory reaction in jennies is induced following AI with frozen-thawed sperm, as a high amount of polymorphonuclear neutrophils (PMN) are observed within the donkey uterus six hours after AI. While PMN appear to select the sperm that ultimately reach the oviduct, two mechanisms, phagocytosis and NETosis, have been purported to be involved in that clearance. Remarkably, sperm interacts with PMN, but the presence of seminal plasma reduces that binding. As seminal plasma is a complex fluid made up of different molecules, including proteins, this study aimed to evaluate how different seminal plasma fractions, separated by molecular weight (<3, 3–10, 10–30, 30–50, 50–100, and >100 kDa), affect sperm–PMN binding. Sperm motility, viability, and sperm–PMN binding were evaluated after 0 h, 1 h, 2 h, 3 h, and 4 h of co-incubation at 38 °C. Two seminal plasma fractions, including 30–50 kDa or 50–100 kDa proteins, showed the highest sperm motility and viability. As viability of sperm not bound to PMN after 3 h of incubation was the highest in the presence of 30–50 and 50–100 kDa proteins, we suggest that both fractions are involved in the control of the jenny’s post-breeding inflammatory response. In conclusion, this study has shown for the first time that specific fractions rather than the entire seminal plasma modulate sperm–PMN binding within the donkey uterus. As several proteins suggested to be involved in the control of post-AI endometritis have a molecular weight between 30 and 100 kDa, further studies aimed at determining the identity of these molecules and evaluating their potential effect in vivo are much warranted.

Alpaca semen quality in relation to different diets

2013 ◽  
Vol 25 (4) ◽  
pp. 683 ◽  
Author(s):  
N. S. Juyena ◽  
J. Vencato ◽  
G. Pasini ◽  
I. Vazzana ◽  
C. Stelletta
Keyword(s):  
Molecular Weight ◽  
Alkaline Phosphatase ◽  
Seminal Plasma ◽  
Total Protein ◽  
Semen Quality ◽  
Sodium Dodecyl ◽  
Sperm Concentration ◽  
Semen Parameters ◽  
Volume Viscosity ◽  
Glutamyl Transferase

The aim of the present study was to evaluate the biochemical composition of seminal plasma, along with semen quality, of alpacas maintained on different diets (hay; hay + pasture grazing; pasture grazing + sheep concentrate; pasture grazing + horse concentrate; Periods 1–4, respectively). Alpacas (n = 5) were fed the four different diets for a period of 6 weeks each. During the period of feeding of each diet, semen was collected using an artificial vagina to determine its volume, viscosity, sperm concentration and sperm motility. Moreover, testicular volume and body condition score were evaluated. Seminal plasma was analysed biochemically to measure total protein, triglyceride, cholesterol, γ-glutamyl transferase, alanine aminotransferase (ALT) and alkaline phosphatase levels. Protein profiles were investigated using one-dimensional sodium dodecyl sulfate–polyacrylamide gel electrophoresis. There was high variability in semen parameters between different males maintained on the same diet. Semen volume increased significantly (P < 0.05) when alpacas were fed diets containing commercial sheep and horse concentrates. In contrast, sperm concentration and motility decreased significantly (P < 0.05) from Period 1 to Period 4. Dietary changes had no effect on viscosity. Significant reductions were seen in triglyceride and cholesterol content, as well as γ-glutamyl transferase, ALT and alkaline phosphatase concentrations, from Period 1 to Period 4. Regardless of experimental period, a wide variation was seen in seminal plasma enzyme concentrations between alpacas, whereas diet had no effect on glucose and total protein concentrations in the seminal plasma. Eight protein bands, with molecular weights ranging from 200 to 14 kDa, were considered in electrophoresis gel after image analysis. Proteins fractions of the 14-kDa (total protein express in md dL–1 with a molecular weight of 14-kDa, TP8) and 21-kDa (total protein express in md dL–1 with a molecular weight of 21-kDa, TP7) bands were not present in all samples of alpaca seminal plasma. There were no significant changes in the concentration of any protein fractions during the four periods. Moreover, the protein fraction of the 60-kDa (total protein express in md dL–1 with a molecular weight of 60-kDa, TP3) band was the most prevalent in all periods. These results demonstrate that there are marked changes in semen quality, as well as some parameters related to the composition of alpaca seminal plasma, that are dependent on diet, which may indicate the need for specific diet formulation to improve reproductive performance. We hypothesise that, in alpacas, the mechanisms underlying the changes in some reproductive traits in response to feeding regimens could be related to changes in the endocrine–gonadal system.

scholarly journals Seminal plasma protein profile based on molecular weight and their correlation with semen quality of Simmental bull

2020 ◽  
Vol 46 (1) ◽  
pp. 20-28
Author(s):  
A. Baharun ◽  
I. Arifiantini ◽  
N. W. K. Karja ◽  
S. Said
Keyword(s):  
Molecular Weight ◽  
Plasma Protein ◽  
Seminal Plasma ◽  
Semen Quality ◽  
Protein Profile ◽  
Sperm Concentration ◽  
Protein Characterization ◽  
Protein Assay ◽  
Frozen Semen

Artificial Insemination Center (AIC) produces frozen semen that was taken from superior bull. The selection of superior bull was based on breeding soundness evaluation. This study is aimed to evaluating the correlation between the molecular weight (MW) of seminal plasma protein and semen quality of Simmental bulls. Semen ejaculates from nine Simmental Bulls, that belong to AIC of Central Java, at Ungaran, were collected by using an artificial vagina. The sub population of sperm in ejaculates was evaluated through macroscopically and microscopically, in which the semen then centrifugated 6500 rpm for 30 minutes. The supernatant was collected and inserted into mini straw and stored in liquid nitrogen container. The concentration of seminal plasma protein was determined by the Bradford method. The Bradford protocol of analysis was suitable with User Guide Coomasize (Bradford) Protein Assay Kit. Thermo Skanlt RE for Multiskan Go Software, 3.2 version was applied for analyzing of the data, and protein characterization used was 1D-SDS-PAGE. The Coomassie Brilliant Blue was used for coloring the gels, and the proteins MW was determined by MW markers. The result demonstrated that there was no significant correlation between protein concentration and semen quality (P>0.05). Protein expressions based on MW were 62-48 kD, 100-71 kD found in this study. The analysis of correlation showed that the proteins with 100-71 kD MW correlated with sperm motility, normal sperm morphology, and sperm concentration. Semen quality in all parameters was significant (P˂0.05 and P>0.01) with 62-48 kD protein MW. The result concluded that seminal plasma protein had a correlation with semen quality and can be used as additional indicator for bull’s candidate selection. 

A Labile CO2-Fixing Enzyme Complex in Spinach Chloroplasts

1972 ◽  
Vol 27 (8) ◽  
pp. 925-932 ◽  
Author(s):  
Bruno Müller
Keyword(s):  
Molecular Weight ◽  
Enzyme Activity ◽  
Phosphoglycerate Kinase ◽  
Gel Chromatography ◽  
Enzyme Complex ◽  
Molecular Weights ◽  
Spinach Chloroplasts ◽  
Reversible Dissociation ◽  
Glyceraldehyde Phosphate Dehydrogenase ◽  
Ribulose Diphosphate Carboxylase

Activity and activation of ribulose 5-phosphate kinase, ribulose diphosphate carboxylase, phosphoglycerate kinase, and the NADP- and NAD-dependent glyceraldehyde phosphate dehydrogenases from isolated diloroplasts were traced in the course of differential ultracentrifugation. Activation of some of these enzymes was carried out by incubation with NADPH2 or ATP. Activity of ribulose diphosphate carboxylase could be increased 2,4-fold by incubation with NADPH2 while phosphoglycerate kinase could not be activated.The degree of activation of the NADP-specific glyceraldehyde phosphate dehydrogenase and ribulose phosphate kinase was correlated with the velocity of sedimentation. Enzymes, which could be activated manifold, sedimented more quickly. Two fractions could be obtained after ultracentri fugation: the sedimented fraction could be highly activated while the fraction in the supernatant could not.It was shown by gel chromatography with Sephadex for the NADP-specific glyceraldehyde phosphate dehydrogenase and ribulose phosphate kinase that the enzyme fractions which could be greatly activated had molecular weights of at least 400 000. In contrast, the enzyme fraction which could not be activated had a molecular weight of 50 000 for the ribulose phosphate kinase and of 240 000 for the glyceraldehyde phosphate dehydrogenases.Results are discussed in the way that a reversible dissociation of a labile enzyme complex takes place between ribulose phosphate kinase, glyceraldehyde phosphate dehydrogenases, and ribulose diphosphate carboxylase. Enzyme activity changes with the state of aggregation. The dissociated enzymes have higher activity than the complexed ones. Dissociation can be performed by ATP and NADPH2, which are assumed to be the physiological regulators, and by unphysiological means as dilution and high salt concentrations.

scholarly journals Modification of Arabinogalactan Isolated from Larix sibirica Ledeb. into Sulfated Derivatives with the Controlled Molecular Weights

Molecules ◽  
2021 ◽  
Vol 26 (17) ◽  
pp. 5364
Author(s):  
Yuriy N. Malyar ◽  
Natalia Yu. Vasilyeva ◽  
Aleksandr S. Kazachenko ◽  
Valentina S. Borovkova ◽  
Andrei M. Skripnikov ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Numerical Optimization ◽  
Sulfamic Acid ◽  
Larix Sibirica ◽  
Process Time ◽  
Reaction Products ◽  
Molecular Weights ◽  
High Sulfur Content ◽  
Natural Polysaccharide ◽  
First Time

The process of sulfation of arabinogalactan—a natural polysaccharide from Larix sibirica Ledeb.—with sulfamic acid in 1,4-dioxane using different activators has been studied for the first time. The dynamics of the molecular weight of sulfated arabinogalactan upon variation in the temperature and time of sulfation of arabinogalactan with sulfamic acid in 1,4-dioxane has been investigated. It has been found that, as the sulfation time increases from 10 to 90 min, the molecular weights of the reaction products grow due to the introduction of sulfate groups without significant destruction of the initial polymer and sulfation products. Sulfation at 95 °C for 20 min yields the products with a higher molecular weight than in the case of sulfation at 85 °C, which is related to an increase in the sulfation rate; however, during the further process occurring under these conditions, sulfation is accompanied by the destruction and the molecular weight of the sulfated polymer decreases. The numerical optimization of arabinogalactan sulfation process has been performed. It has been shown that the optimal parameters for obtaining a product with a high sulfur content are a sulfamic acid amount of 20 mmol per 1 g of arabinogalactan, a process temperature of 85 °C, and a process time of 2.5 h.

Low molecular weight components in bovine semen diffusate and their effects on motility of bull sperm

1994 ◽  
Vol 6 (2) ◽  
pp. 165 ◽  
Author(s):  
N al-Somai ◽  
R Vishwanath ◽  
P Shannon ◽  
PC Molan
Keyword(s):  
Molecular Weight ◽  
Sperm Motility ◽  
Seminal Plasma ◽  
Detrimental Effect ◽  
Gel Permeation Chromatography ◽  
Low Molecular Weight ◽  
Small Molecular Weight ◽  
Bovine Semen ◽  
Molecular Weights ◽  
Sperm Survival

Dialysis of diluted semen before cryopreservation is beneficial to sperm survival. This is due to removal of low molecular weight components from seminal plasma that are damaging to sperm. The apparent molecular weights (M(r)) of these components range between 1000 and 12,000 as estimated by gel permeation chromatography and electrophoresis. The detrimental effect on sperm motility is greatest with the components of M(r) between 5000 and 12,000 and with those of M(r) < 1500. Their effect on sperm motility was dependent on concentration. The small molecular weight components were derived from the high molecular weight components of seminal plasma through disaggregation under prescribed conditions.

scholarly journals Determination of enzyme activity in rabbit seminal plasma and its relationship with quality semen parameters

2015 ◽  
Vol 23 (4) ◽  
pp. 247 ◽  
Author(s):  
M.P. Viudes de Castro ◽  
L. Casares-Crespo ◽  
A. Monserrat-Martínez ◽  
J.S. Vicente
Keyword(s):  
Enzyme Activity ◽  
Seminal Plasma ◽  
Semen Quality ◽  
Sperm Concentration ◽  
Significant Negative Correlation ◽  
Semen Parameters ◽  
Male Age ◽  
Head Displacement ◽  
The Status ◽  
Positive Correlations

<p>The objective of this study was to determine rabbit seminal plasma enzyme activity. Furthermore, correlations between semen parameters and enzyme activity and male age were examined. The study was performed using 17 New Zealand White males from 5 to 9 mo old. Overall, 252 semen samples were collected from bucks from May to September. Semen characteristics were analysed and the seminal plasma was obtained by centrifugation. The activities of alanyl aminopeptidase (APN), aspartate transaminase (AST), alanine aminotransferase (ALT), γ-glutamyl transpeptidase (GGT), lactate dehydrogenase (LDH) and alkaline phosphatase (ALKP) in the seminal plasma fluid were measured. Significant differences between males were found in APN, GGT, LDH, ALKP and ALT activities (P&lt;0.05). No significant differences between enzyme activity and male age were found. We also observed significant positive correlations between male age and sperm concentration (r=0.26), progressive motility (r=0.17) and amplitude of lateral head displacement (r=0.21), and negative ones between male age and average path velocity (r=–0.56), velocity of the sperm head along its actual curvilinear path (r=–0.61), straight line velocity (r=–0.50), linearity index (r=–0.13), and cytoplasmic droplet (r=–0.33). Furthermore, a significant negative correlation between APN activity and the status of the acrosome (r=–0.20) and significant positive correlations between APN activity and the sperm abnormalities (r=0.21), GGT activity and sperm concentration (r=0.34) and the status of the acrosome (r=0.31), and ALKP activity and sperm concentration were observed (r=0.41). In our study, APN and GGT seem to be the most predictive enzymes for rabbit semen quality.</p>

Green-solvent processable semiconducting polymers applicable in additive-free perovskite and polymer solar cells: molecular weights, photovoltaic performance, and thermal stability

2018 ◽  
Vol 6 (14) ◽  
pp. 5538-5543 ◽  
Author(s):  
Junwoo Lee ◽  
Tack Ho Lee ◽  
Mahdi Malekshahi Byranvand ◽  
Kyoungwon Choi ◽  
Hong Il Kim ◽  
...  
Keyword(s):  
Thermal Stability ◽  
Molecular Weight ◽  
Solar Cells ◽  
Polymer Solar Cells ◽  
Photovoltaic Performance ◽  
Semiconducting Polymers ◽  
Green Solvent ◽  
Molecular Weights ◽  
First Time

In this study, we demonstrated the effects of the molecular weight (MW) of a green processable polymer (asy-PBTBDT) on its photovoltaic performance and thermal stability for the first time.

Investigation by HPLC of the Catabolism of Recombinant Tissue Plasminogen Activator in the Rat

1988 ◽  
Vol 60 (01) ◽  
pp. 107-112 ◽  
Author(s):  
Roy Harris ◽  
Louis Garcia Frade ◽  
Lesley J Creighton ◽  
Paul S Gascoine ◽  
Maher M Alexandroni ◽  
...  
Keyword(s):  
Molecular Weight ◽  
Plasminogen Activator ◽  
Tissue Plasminogen Activator ◽  
Half Life ◽  
Recombinant Tissue Plasminogen Activator ◽  
Rat Plasma ◽  
High Molecular Weight Complex ◽  
Molecular Weights ◽  
Major Activity ◽  
Tissue Plasminogen

SummaryThe catabolism of recombinant tissue plasminogen activator (rt-PA) was investigated after injection of radiolabelled material into rats. Both Iodogen and Chloramine T iodination procedures yielded similar biological activity loss in the resultant labelled rt-PA and had half lives in the rat circulation of 1 and 3 min respectively. Complex formation of rt-PA was investigated by HPLC gel exclusion (TSK G3000 SW) fractionation of rat plasma samples taken 1-2 min after 125I-rt-PA injection. A series of radiolabelled complexes of varying molecular weights were found. However, 60% of the counts were associated with a single large molecular weight complex (350–500 kDa) which was undetectable by immunologically based assays (ELISA and BIA) and showed only low activity with a functional promoter-type t-PA assay. Two major activity peaks in the HPLC fractions were associated with Tree t-PA and a complex having a molecular weight of ̴ 180 kDa. HPLC fractionation to produce these three peaks at various timed intervals after injection of 125I-rt-PA showed each to have a similar initial rate half life in the rat circulation of 4-5 min. The function of these complexes as yet is unclear but since a high proportion of rt-PA is associated with a high molecular weight complex with a short half life in the rat, we suggest that the formation of this complex may be a mechanism by which t-PA activity is initially regulated and finally cleared from the rat circulation.

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