Biomedical subjects

M Fasinski

Publications and source records attributed to M Fasinski.

4 recordsLinked to original sources

PubMed1992

Bovine early pregnancy factor (EPF) activity dependent on a 67-kDa polypeptide.

Maternal bovine EPF activity can be reduced to one single polypeptide enriched and identified from serum of cows in early pregnancy. The relative molecular weight of this active polypeptide was estimated at 67 kDa. This bovine EPF was labelled by 125I and peroxidase. In parallel investigations of non-pregnant animals a 67-kDa polypeptide was additionally identified in the last purification step, but without EPF activity in the rosette inhibition test. This indicated occurrence of an inactive pre-compound (or carrier protein) of the EPF in the non-pregnant state. On pre-incubation of lymphocytes with EPF analogues (inactive polypeptide from nonpregnancy serum) EPF retained its optimal activity, its lymphocyte receptors being unaffected. Monoclonal antibodies produced against HPLC-enriched EPF were reactive to the 67-kDa polypeptide in pregnancy material as well as in nonpregnancy material and were not able to differentiate between 'pregnant' and 'nonpregnant'. A mouse anti-EPF serum produced against highly purified EPF isolated from SDS PAGE showed reactivity only against the 67-kDa polypeptide of pregnancy serum but not against that of non-pregnancy serum. This is the first evidence for a difference in antigenic determinants of the two 67-kDa proteins found in pregnancy and non-pregnancy serum. Furthermore, a second higher molecular weight protein could be identified by this antiserum in pregnancy and non-pregnancy serum.

Animals↗

PubMed1989

[The detection of in vitro capacitation of bull sperm by heparin treatment].

The capacitating effect of heparin upon spermatozoa from original and deep-frozen semen was characterised, using new methods for detection of inducible acrosomal reaction and heparin-mediated sperm aggregation, and was compared with frequently used capacitation by media of high ion strength. Heparin treatment was undertaken also by means of two culture media, "defined medium" (DM) and TCM 199, with 10% fetal calf serum. Higher motility was maintained by means of 10 I.U. of heparin/ml (= 77 micrograms/ml) which also proved helpful in achieving higher capability of inducible acrosomal reaction, as compared to pretreatment, using media of high ion strength. This applied to both fresh and deep-frozen sperm. The highest level of inducible acrosomal reaction was achieved after 2 hours of heparin action on fresh sperm and 30 minutes of action on deep-frozen sperm. That highest value was at its maximum, when TCM with 10% fetal calf serum had been used. This was the medium, after all, in which photometrically recorded aggregation of motile spermatozoa was at its fastest rate, reaching its maximum after about 60 minutes. The photometrically recorded activated motility of spermatozoa occurred more frequently in TCM, as compared to DM. Preparation of bull sperm in TCM 199 with fetal calf serum and heparin may be recommended as an effective and time-saving method for in vitro capacitation.

Animals↗

PubMed1989

[Characterizing the in vitro capacitation of motile bull sperm by the hyamine-induced acrosome reaction].

Described in this paper is a method to characterize capacitation of bull spermatozoa. Involved is the action of 0.005 per cent of hyamine 2389 on selected motile cells at 37 degrees C for 30 minutes to detect the number of spermatozoa capable of acrosomal reaction. The latter amount results from the percentual difference between acrosome-free spermatozoa prior to hyamine action and those after hyamine action. Acrosome-free spermatozoa can be easily identified by means of simple staining under optical light microscopy. The value thus obtained is related to the amount of spermatozoa in progressively motile in the unselected original suspension and is called "Percentage Acrosome Reaction Induction" (% ARI). The ARI capacitation-specific acts: Only small amounts of acrosome-free spermatozoa are recordable among incapacitated spermatozoa or from spermatozoa incubated in the presence of decapacitation factors (1.9 or 2.3 per cent, respectively). On the other hand, clearly increased values of 17.6 ot 26.1 per cent are obtainable from treatments with capacitation effect (high ionic strength or addition of heparin). The method is claimed to be suitable for adequate choice of conditions for improved capacitation and in vitro fertilization as well as for elucidation of specific individual differences in capacitation.

Acrosome↗

PubMed1989

[A simple method for the quantitative determination of sperm aggregation].

Described in this paper are 2 methodological variants for photometric recording of sperm aggregation. Heparin was used to induce aggregation. One of the working principles was related to aggregation-associated alteration of turbidity in an agitated cell suspension, measured by the KZM-1 coagulation-time meter, in analogy to measurement of thrombocyte aggregation. The 2nd variant was based on variation of cloudiness due to sedimentation of aggregates in a non-agitated suspension. Both methods provided equally valid information. They characterised both intensity of aggregation as a combined effect of the number and size of aggregates as well as the rate of aggregation. Photometric recording has proved to be an objectivated method for quantitative assessment of aggregation. Its use is proposed for studies into capacitation or immunological response of spermatozoa.

Animals↗