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Biomedical subjects

E F Graham

Publications and source records attributed to E F Graham.

At least 37 records · Page 2Linked to original sources
PubMed1987

Factors affecting the removal of low-molecular-weight fractions (LMWF) from egg yolk and seminal plasma in extended semen by dialysis: effect on post-thaw sperm survival.

Three factors affecting dialysis of bovine semen were studied. These factors were (1) dialysis rates of egg yolk, seminal plasma, and glycerol, (2) temperature (37 degrees C, 5 degrees C, and while cooling from 37 to 5 degrees C), and (3) dialysis ratios between retentate and dialysate (1:1, 1:10, 1:20, 1:50, and 1:100). Ninety percent of the low-molecular-weight fraction (LMWF) from seminal plasma, egg yolk, and glycerol was removed from the retentate in a 2-hr period at 5 degrees C, and only slight changes were detected after the third hour of dialysis. Temperature affected dialysis and was faster at 37 degrees C. It was also found that a 1:20 dialysis ratio was sufficient to obtain 90% clearance of the LMWF. The effect of sperm dilution ratio, dialysis ratio, and exchange of the LMWF from egg yolk and/or seminal plasma for buffer systems was also studied. An improvement in post-thaw motility of spermatozoa (P less than 0.05) was obtained when the LMWF from both seminal plasma and egg yolk were replaced. A third experiment was conducted to study the effect of different combinations between the buffer systems, TEST and Na citrate, in the dialysate. The results indicated that a 1:1 combination of iso-osmotic solutions (320-325 mOsm/Kg, pH 7.0) between these two buffers, with 5% glycerol (v/v), yielded significant (P less than 0.05) sperm post-thaw motility as compared with the individual use of TEST-glycerol or Na citrate-glycerol. Dialyzed samples also yielded sperm post-thaw motility higher than that of the nondialyzed samples. Colloidal materials in the dialysate did not affect survival of spermatozoa.

Animals↗
PubMed1987

Dialysis of bovine semen and its effect on fresh and freeze-thawed spermatozoa.

The effect of the removal of the low-molecular-weight fraction (LMWF, less than 12,000-14,000 Da) from the seminal plasma present in extended semen by dialysis and by centrifugation (1,376g for 20 min at 5 degrees C) were compared with the current methods of freezing bovine semen. Significantly higher sperm post-thaw motility (P less than 0.05) was obtained in the dialyzed samples than with the other two methods. The appropriate time and temperature for dialysis of semen was also studied. Semen aliquots were dialyzed (1:50, retentate:dialysate) for 30 min, 1 or 2 hr at 5 degrees C, and during the cooling process from 37 to 5 degrees C over a 2-hr period. Superior sperm motility (P less than 0.05) in prefreeze and post-thawed samples was observed when semen was dialyzed for 1 or 2 hr during the cooling process as compared with that of semen dialyzed at 5 degrees C. A third experiment was conducted to establish the effect of the use of dialysis bags of different molecular weight cutoffs (MWCO) on sperm motility. Semen samples were dialyzed (1:50) during the cooling process in dialysis bags of 1,000, 3,500, 6,000-8,000, 12,000-14,000, 25,000, and 50,000 MWCO. No statistical differences (P greater than 0.05) in sperm post-thaw motility were found after evaluation of the number of cells that passed through the Sephadex filter and all the dialyzed values obtained were significantly (P less than 0.05) superior to the results obtained with no dialysis.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1986

The effect of dialysis of extended ram semen prior to freezing on post-thaw survival and fertility.

The effect of dialysis on extended ram semen prior to cryopreservation was studied. Techniques were developed to improve post-thaw recovery of dialyzed semen and a fertility trial was used to evaluate the viability of dialyzed and frozen semen. Dialysis prior to freezing was shown to increase post-thaw recovery of motile cells and percentage of cells passing through a Sephadex filter. Freezing semen in pellets on dry ice was superior to freezing in French straws. Pellets were thawed in an aluminum thaw block at 42 to 45 degrees C before insemination of progestagen-PMSG synchronized ewes. Double inseminations were made at 12-hr intervals. Natural service of synchronized ewes was also made at 12-hr intervals as a control. There was no significant difference (P greater than 0.05) in fertility between naturally serviced ewes (44.4%) and ewes inseminated with frozen semen (44.7%).

Animals↗
PubMed1986

The effects of nonpenetrating cryoprotectants added to TEST-yolk-glycerol extender on the post-thaw motility of ram spermatozoa.

The effects of adding five different concentrations of 17 polymeric compounds to TEST-yolk-glycerol extender on ram spermatozoa survival was studied. These were Aquacide (I, II, and III); dextran (0.8-1.6, 1.9, 15-20, 70, and 200-300 kDa); three types of Dri-Sweet; hydroxyethyl starch; methylcellulose, polyethylene glycol, polyvinyl alcohol, polyvinyl pyrrolidone, and Supercol 912. All the compounds tested except the Dri-Sweet compounds and hydroxyethyl starch significantly (P less than 0.05) decreased percentages of motile cells in unfrozen samples. The use of dextran (0.8-1.6 kDa; hydrolyzed dextran separated by ethanol) and Aquacide II significantly (P less than 0.05) increased post-thaw motility of spermatozoa frozen in pellets. Dextran (15-20 kDa), dextran (0.8-1.6 kDa), Aquacide II, and hydroxyethyl starch significantly (P less than 0.05) increased the percentages of post-thaw motility of ram spermatozoa frozen in the presence of glycerol and egg yolk.

Animals↗
PubMed1983

Effects of various degrees of supercooling and nucleation temperatures on fertility of frozen turkey spermatozoa.

The relative roles of the degree of supercooling and nucleation temperatures on turkey sperm cell survival and fertilizing capacity during a freeze-thaw cycle were investigated. Basically, the higher the degree of supercooling, which produced lower spontaneous nucleation temperatures during the freezing process, the worse the sperm cells survived, in the present study. When induced nucleation was applied, which eliminated the high degree of supercooling, an improvement in sperm cell survival was noted. During the fertility trial, it was shown that in treatment 3 (induced nucleation) the fertility was higher (P less than 0.05) than in treatment 2 (spontaneous nucleation). In general, the degree of supercooling prior to freezing is an important variable and should be considered very seriously during the overall freezing process.

Animals↗
PubMed1983

Alteration of seminal proteins during freeze-drying of bovine semen.

Bovine semen in TEST-yolk extender was frozen, freeze-dried to 50, 25, 12, 6, and less than 2% residual moisture, and stored at -196 degrees C. The freeze-dried semen was rehydrated, sampled for protein analysis, and used to inseminate cattle. Agar gel electrophoresis revealed no moisture was reduced to less than 6%. At that point, the percent of cationic proteins and neural proteins decreased with a concurrent increase in anionic migrating proteins. Immunodiffusion data with antisera against spermatozoa and seminal plasma revealed no difference in formation of precipitin lines if residual moisture was at least 6%. However, semen freeze-dried to 2% residual moisture was modified antigenically as certain precipitin lines were lost, new lines appeared, and concentration of other seminal antigens decreased. The absence of fertility with semen freeze-dried to 2% residual moisture is hypothesized to 2% residual moisture is hypothesized to be from alteration of the tertiary structure of certain essential seminal proteins.

Animals↗
PubMed1982

Development of extender and techniques for frozen turkey semen. 1. Development.

An extender for turkey semen with a frozen-thawed recovery of greater than or equal to 50% motile spermatozoa and a vigorous swirl was developed. The effect of glucose, a comparison of 10 different sugars in the extender, the use of sodium acetate and potassium acetate, the ratio of dimethylsulfoxide to ethylene glycol, the percent total cryoprotectant, equilibration time, and osmotic pressure were all tested. Replacement of approximately half of the extender with an iso-osmolar glucose solution yielded higher percentages of motile spermatozoa with both fresh and frozen-thawed semen samples. Replacement of glucose with other carbohydrates did not enhance recovery of frozen-thawed semen. The proportion of sodium and potassium acetates to TESNaK2 in the extender had no effect on motility. Approximately equal proportions of ethylene glycol to dimethylsulfoxide, with a final concentration of 11.2% after dilution, produced at or near optimal recovery of spermatozoal motility after freezing. Twenty to 90 min equilibration times before freezing yielded significantly higher recovery of motile sperm postthaw than shorter periods. A wide range of extender osmotic pressures were compatible with recovery of spermatozoal motility postthaw. The final extender consisted of 4.70 g sodium acetate, 3.39 g potassium acetate 9.23 g TES, .353 g sodium hydroxide, .492 g potassium hydroxide, 32.00 g glucose, H2O q.s. 1000 ml, 370 mOsm/kg, pH 7.2. Ethylene glycol and dimethylsulfoxide were added (1:1 ratio) for a final concentration of 11.2% cryoprotectant after dilution. Semen was held undiluted for 6 min, extended 1:4 at 22 C, and equilibrated for 30 min at 0 C before pellet freezing.

Animals↗
PubMed1982

Development of extender and techniques for frozen turkey semen. 2. Fertility trials.

The effects of cryoprotectant concentrations, redilution, centrifugation, and dialysis techniques on fertility of fresh and frozen-thawed turkey semen were tested. Fresh extended semen with final concentrations of 4.8, 5.6, 9.6, and 11.2% cryoprotectant (dimethylsulfoxide and ethylene glycol, 1:1 ratio) showed a significant (P less than .05) decrease in fertility when compared to semen without cryoprotectant. Fertility of fresh semen extended with 4.8% cryoprotectant when centrifuged or rediluted and centrifuged, was unaffected when compared to cryoprotectant treated semen. Fertility of frozen-thawed semen after redilution and centrifugation was low, but samples rediluted one part semen for two parts extender yielded significantly (P less than .05) higher fertility than semen rediluted 1:1. Fresh semen extended with 11.2% cryoprotectant showed a highly significant (P less than .01) decrease in fertility despite dialysis, while semen with 9.6% cryoprotectant showed no difference when compared to dialyzed semen without cryoprotectant. Fertility of frozen-thawed semen dialyzed against blood plasma (approximately pH 7.9) and blood plasma adjusted with TES2 to pH 7.4 or pH 7.2 was not significantly different. Fertility of frozen-thawed semen dialyzed for 30 min was significantly higher (P less than .01) than semen dialyzed for 15 min or 0 min. Frozen-thawed semen dialyzed with a semen to dialysate ratio of 1:15 and 1:24 supported significantly higher (P less than .01) fertility than at 1:50 ratio. The AGPT3 dialysate yielded significantly higher (P less than .01) fertility than blood plasma adjusted to pH 7.2 with TES.

Animals↗
PubMed1981

The in vitro effect of boar semen on the contractility of rat uteri.

Contradictory reports in the literature provoked the investigation of a possible oxytocic effect of boar semen on in vitro rat uterus preparations. When fresh boar seminal plasma (SP), dialyzed seminal plasma (DSP) or a filtrate of acetone-extracted seminal plasma (AE) were added to uterine preparations, both the frequency and force of spontaneous contractile activity tended to be depressed. When the uterine preparations were primed with oxytocin (1.6 USP units/100 ml bathing solution), treatment with SP and AE resulted in an increased frequency of contraction but no increase in force. DSP increased neither the frequency nor force of contractions. A solution of salts and organic compounds that approximated the small molecular or dialyzable components of boar semen ("synthetic boar seminal plasma" or BS) affected contractility in a manner similar to SP and AE. When the rat uterus was bathed in a low calcium physiological salt solution, none of the seminal treatments (SP, DSP, AE, or BS) were capable of initiating a contraction. Further, the force of carbachol-induced contractions in these uterine preparations was markedly depressed by these seminal treatments. In contrast, treatment with oxytocin nearly doubled the contractile force. The depressant effect of SP, DSP and AE on both spontaneous and carbachol-induced contractions appeared to be irreversible. Mineral analysis of SP, DSP, AE, BS and the low calcium salt solution showed the SP, AE and BS contained amounts of Ca(2+) and Mg(2+) which could have altered the uterine contractility. In addition, a non-dial-yzable factor in SP appeared to have a similar effect.

Journal Article↗
PubMed1979

Assay of frozen boar semen with Sephadex filtration.

The suitability of filtration of frozen boar semen through Sephadex G-15-120 as a viability assay was investigated. Semen thawed on a hot plate at 38 degrees C was counted with a Coulter Counter before and after filtering through Sephadex columns with 0.1 M sodium citrate as flushing medium. More spermatozoa passed through the column when the temperature of the flushing medium was elevated from room temperature to 37 degrees C and added with 5 mM caffeine (12.3 vs. 22.8% p<0.01). The use of caffeine and 37 degrees C flushings produced a filtrate containing spermatozoa with 89+/-4% motility and 97.6+/-1.6% normal acrosome ridges with the use of frozen semen from 5 different boars. The repeatability was +/-7.3%. The unfiltered samples were judged to contain 45 +/- 8% motile spermatozoa and 66.6 +/- 7.1% spermatozoa with normal acrosome ridges. Filtering of frozen boar semen through Sephadex is proposed as a rapid, objective assay combining the benefits of differential counting of normal acrosome ridges and motility determination.

Journal Article↗
PubMed1979

Effect of alpha-chlorohydrin on epididymal sperm and epididymal plasma in swine.

Investigation of the contraceptive mechanism of alpha-chlorohydrin was done by analyses of epididymal plasma and certain epididymal sperm characteristics after oral administration of 0, 5, 10 or 30 mg/kg, day of the drug to boars for 15 days. Water resorption in caput epididymidis was slightly decreased in all treatment groups. Sodium, potassium, chloride, glycerylphosphorylcholine levels, and seminal antigens in epididymal plasma were not altered significantly by 10 or 30 mg/kg of the drug. The boars on 5 mg/kg exhibited significantly elevated sodium, potassium, or chloride values in various segments. Motility was significantly lower on corpus and proximal cauda epididymal spermatozoa from alpha-chlorohydrin treated boars. Only boars receiving 30 mg/kg exhibited impaired sperm motility in the distal cauda. The movement of the cytoplasmic droplets to the distal position was retarded in boars on the two highest dose levels. The results suggest that the contraceptive effects of alpha-chlorohydrin in the boar is probably not mediated via an impaired epididymal function.

Animals↗