Distribution of variance associated with measurement of post-thaw function in ram sperm

1995 ◽  
Vol 7 (1) ◽  
pp. 129 ◽  
Author(s):  
AY Edward ◽  
DP Windsor ◽  
IW Purvis ◽  
LG Sanchez-Partida ◽  
WM Maxwell
Keyword(s):  
Total Variation ◽  
Semen Analysis ◽  
Amidase Activity ◽  
Ram Sperm ◽  
Semen Characteristic ◽  
The Way

Post-thaw characteristics of ram semen frozen as pellets were assessed using biochemically (amidase activity) or motility-based (Hamilton Thorn Motility Analyzer) techniques. The total variation associated with each semen characteristic measured was partitioned between rams (5), ejaculates within rams (5), pellets within ejaculates (5) and within pellets (2). A variety of variance distributions were observed for the characteristics measured. Of the 18 post-thaw characteristics examined, 10 had > 50% of variance distributed between within-ejaculate components. This has important implications for the way in which such measurements may be used in post-thaw semen analysis.

55 EFFECT OF TREHALOSE FOR FROZEN–THAWED RAM SPERM MOTILITY PARAMETERS

2015 ◽  
Vol 27 (1) ◽  
pp. 120
Author(s):  
M. M. Toishibekov ◽  
M. T. Jazkbayev ◽  
B. B. Molzhigitov
Keyword(s):  
Sperm Motility ◽  
Semen Analysis ◽  
Sperm Concentration ◽  
Egg Yolk ◽  
Computer Assisted ◽  
Progressive Motility ◽  
Indigenous Sheep ◽  
Standard Tool ◽  
Freezing Curve ◽  
Ram Sperm

Computer-assisted sperm analysers have become the standard tool for evaluating sperm motility because they provide objective results for thousands of mammalian spermatozoa. Ram semen was collected using electro-ejaculation from 10 adult rams of Chingizskaya indigenous sheep breed. Motility was determined using computer-automated semen analysis (Hamilton Thorne Motility Analyzer, Beverly, MA, USA). Trehalose solution (0.375 M) was added to Tris-buffered saline solution to give the following trehalose extenders: 25, 50, 75, and 100% (vol:vol), and analysed for motility using computer-automated semen analysis. The sperm pellets were resuspended at 24°C in cooling extender – trehalose extenders of each concentration containing 5% egg yolk. The diluted semen was cooled to 5°C within 2 h. The semen was then further diluted 1 : 1 with freezing extender – each trehalose extender containing 1.5% glycerol to obtain a sperm concentration of 2.0 × 108 cells mL–1 – and then loaded into 0.5-mL straws. Straws were frozen using a programmable freezer with a freezing curve of 5°C to –5°C at 4°C per min, –5°C to –110°C at 25°C per min, and –110°C to –140°C at 35°C per min, and then the straws were plunged into liquid nitrogen for storage. Frozen samples were thawed in a 37°C water bath for 30 s and analysed for motility using computer-automated semen analysis. Statistical analyses were performed with a Student's test. The fresh semen samples showed the next results: motility 88.3 ± 2.4%, progressive motility 26.8 ± 6.9%, and progressive velocity 61.9 ± 4.2 μm s–1. Motility of the frozen-thawed spermatozoa was 63.6 ± 2.9% (25% trehalose), 55.6 ± 5.2% (50%), 32.4 ± 4.7% (75%), and 23.6 ± 3.2 (100%). Progressive motility was 15.6 ± 3.9% (25%), 13.7 ± 3.7% (50%), 4.5 ± 1.3% (75%), and 5.2 ± 1.3% (100%). Progressive velocity was 93.5 ± 8.3 μm s–1 (25%), 85.4 ± 8.1 μm s–1 (50%), 65.7 ± 6.1 μm s–1 (75%), 35.2 ± 3.3 μm s–1 (100%). Motility of the frozen-thawed spermatozoa significantly decreased with increasing concentrations of trehalose in the extender (P < 0.05). These preliminary studies showed that further research is needed of use trehalose for ram spermatozoa cryoconservation.

97 COMPARISON OF MOTILITY AND ACROSOMAL INTEGRITY OF ELECTRO-EJACULATED AND EPIDIDYMAL RAM SPERM AFTER EXPOSURE TO VARIOUS ANISOSMOTIC SOLUTIONS, CRYOPROTECTIVE AGENTS, AND TEMPERATURES

2008 ◽  
Vol 20 (1) ◽  
pp. 129
Author(s):  
O. Varisli ◽  
C. Uguz ◽  
C. Agca ◽  
J. Green ◽  
Y. Agca
Keyword(s):  
Sucrose Solution ◽  
Semen Analysis ◽  
Chilling Stress ◽  
Molecular Probes ◽  
Current Data ◽  
Epifluorescence Microscopy ◽  
Computer Assisted ◽  
Cryoprotective Agents ◽  
Ram Sperm ◽  
Acrosomal Integrity

Effective ram sperm cryopreservation protocols, which would yield acceptable lambing rates following AI, are currently lacking. The objectives of the current studies were to compare the effects of various anisosmotic conditions, cryoprotective agents (CPA), and temperatures on the motility and acrosomal integrity of electro-ejaculated and epididymal ram sperm. Three experiments were conducted. In Experiment 1, electro-ejaculated and epididymal ram sperm were exposed to 75, 150, 225, 600, 900, and 1200 mOsm kg–1 sucrose solutions, held for 5 min, and then returned to isosmotic conditions. Motility characteristics of sperm during exposure to each anisosmotic solution and after return to isosmotic conditions were determined. In Experiment 2, electro-ejaculated and epididymal ram sperm were exposed to 1 m glycerol (Gly), dimethyl sulfoxide (DMSO), ethylene glycol (EG), or propylene glycol (PG) for 5 min and then returned to isosmotic conditions. Motility characteristics of sperm samples during exposure to each CPA solution and after return to isosmotic conditions were determined. In Experiment 3, effects of various temperatures on motility characteristics of electro-ejaculated and epididymal ram sperm were determined after exposure to three different sub-physiologic temperatures (4, 10, and 22�C) for 30 min and subsequently return to 37�C. A computer-assisted semen analysis system was used to determine sperm motility characteristics. Epifluorescence microscopy was used to determine sperm acrosomal integrity after fixing and staining with Alexa Fluor-488-PNA (Molecular Probes, Eugene, OR, USA). The data were analyzed by ANOVA by using SAS (SAS Institute, Inc., Cary, NC, USA). The motility of electro-ejaculated ram sperm was significantly more affected by anisosmotic stress than was that of epididymal ram sperm (P < 0.05). While anisosmotic stress had no effect on acrosomal integrity of epididymal ram sperm, there was a significant reduction in acrosomal integrity for electro-ejaculated ram sperm after the addition and removal of a 75 mOsm sucrose solution. The abrupt addition and removal of 1 m Gly, DMSO, EG, or PG had no effect on the motility and acrosomal integrity of epididymal ram sperm (P > 0.05). However, there was a slight decrease in acrosomal integrity for electro-ejaculated ram sperm after exposure to 1 m Gly, DMSO, or EG (P > 0.05). Both epididymal and electro-ejaculated ram sperm exhibited temperature-dependent loss of motility and acrosomal integrity (P < 0.05). However, electro-ejaculated ram sperm were more sensitive to chilling stress than were epididymal sperm (P < 0.05). In conclusion, current data overall suggest that while epididymal ram sperm are extremely resilient to various cryobiologically relevant stress conditions, ejaculated ram sperm demonstrate greater sensitivity to such stressors. These current findings should be taken into account when handling and developing cryopreservation protocols for ejaculated and epididymal ram sperm.

Optimization of a continuous real-time computerized semen analysis system for ram sperm motility assessment, and evaluation of four methods of semen preparation

1996 ◽  
Vol 8 (2) ◽  
pp. 219 ◽  
Author(s):  
WV Holt ◽  
MJ Palomo
Keyword(s):  
Real Time ◽  
Semen Analysis ◽  
Computer Assisted ◽  
Assessment And Evaluation ◽  
Analysis System ◽  
Set Up ◽  
Assisted Analysis ◽  
Ram Sperm ◽  
Preparation Techniques

Sampling conditions that affect the biological validity of computer-assisted analysis of ram sperm motion were examined using a continuous real-time computerized semen analysis system (Hobson Sperm Tracker). Search radius (SR, 10 settings) and minimum track point (MTP, 10 settings) were varied factorially to evaluate their effects on the inclusion of sperm subpopulations within derived datasets. Low SR (< 12 microns) or high MTP values (> 26 frames) precluded measurements of rapidly moving cells, whereas high SR (> 17 microns) and low MTP settings (< 22 frames) led to erroneous tracking and poor data quality. Suitable settings for these set-up parameters were derived and tested for their biological consistency with four methods of preparing ram semen for computerized analysis. The preparation techniques tested were: centrifugation through sucrose-based Ficoll and Percoll media, a swim-up technique and simple dilution in Tyrode's media. The 'selective' Percoll and swim-up methods generated sperm populations with significantly higher linear velocities and a lower tendency to deviate from linear trajectories than from either the Ficoll method or dilution technique. Deleterious effects of centrifugation were evident, particularly on sperm survival in vitro over several hours. It is concluded that computer-assisted semen analysis provides useful information about the behaviour of ram spermatozoa in vitro, but the measurement conditions must be defined carefully.

49 EFFECT OF ADDING Trolox C AND ASCORBIC ACID TO RAM SPERM BEFORE CRYOPRESERVATION ON THE MOTILITY AND BINDING CAPABILITY

2016 ◽  
Vol 28 (2) ◽  
pp. 154 ◽  
Author(s):  
J. Costa ◽  
W. Lima ◽  
E. Moraes ◽  
P. Sousa ◽  
L. Ramon ◽  
...  
Keyword(s):  
Ascorbic Acid ◽  
Sperm Quality ◽  
Semen Analysis ◽  
Egg Yolk ◽  
Computer Assisted ◽  
Beneficial Effects ◽  
C 30 ◽  
Motility Of Sperm ◽  
Ram Sperm ◽  
Trolox C

Different antioxidants have been tested to improve sperm quality, but distinct and consistent beneficial effects are lacking. The objective of this study was to evaluate whether the addition of different concentrations of the antioxidant Trolox C and ascorbic acid before cryopreservation could improve the binding of sperm to chicken egg perivitelline membrane (PM) after cryopreservation. Three ejaculates of ram were split and diluted with Tris egg yolk diluent to a final concentration of 200 × 106 cells mL–1, and after, the ejaculates were divided into 5 tubes. Each tube received one of the following antioxidants: control, no antioxidant; 200 μM of Trolox C; 300 μM of Trolox C; 0.05% of ascorbic acid; and 0.25% of ascorbic acid. The samples were cooled to 5°C/2 h, packaged into 0.5-mL straws, and frozen in static LN vapor for 15 min before being plunged into LN. Straws were thawed (37°C/30 s). The motility was determined using computer-assisted semen analysis. For PM binding test, PM was put in tubes with 1 mL of TALP and inseminated with 50 000 sperm. The PM and sperm were incubated for 90 min at 37°C in an atmosphere of 5% CO2 in air, and 20 min before the end of the incubation time, 10 μL of Hoechst 33342 was added in each treatment. After each PM was washed 5 times in TALP, placed under the coverslip on a slide, and evaluated by fluorescence microscopy at 400×. Spermatozoa were counted in 6 random fields of each piece of PM. Percentage data were transformed using arcsine prior analysis. Treatment differences were determine by analysis of variance and Tukey test. The total and progressive motility of sperm treated with 0.25% ascorbic acid Trolox C was higher (64.5 and 45%) than 100 μM of Trolox C (61.9 and 42.6%) and 200 μM of Trolox C (64.3 and 46.7%) and control (59.8 and 39.6%; P < 0.05), respectively. The binding test was higher when using 0.25% of ascorbic acid (155.73 cells; P < 0.05) compared with other treatments. Addition of 0.25% ascorbic acid to ram sperm before cryopreservation improved cell cryosurvival rates.

scholarly journals Riemann-Stieltjes Integral

2016 ◽  
Vol 24 (3) ◽  
pp. 199-204 ◽  
Author(s):  
Keiko Narita ◽  
Kazuhisa Nakasho ◽  
Yasunari Shidama
Keyword(s):  
Total Variation ◽  
Bounded Variation ◽  
Stieltjes Integral ◽  
Real Numbers ◽  
Basic Properties ◽  
Finite Sequences ◽  
The Way

Abstract In this article, the definitions and basic properties of Riemann-Stieltjes integral are formalized in Mizar [1]. In the first section, we showed the preliminary definition. We proved also some properties of finite sequences of real numbers. In Sec. 2, we defined variation. Using the definition, we also defined bounded variation and total variation, and proved theorems about related properties. In Sec. 3, we defined Riemann-Stieltjes integral. Referring to the way of the article [7], we described the definitions. In the last section, we proved theorems about linearity of Riemann-Stieltjes integral. Because there are two types of linearity in Riemann-Stieltjes integral, we proved linearity in two ways. We showed the proof of theorems based on the description of the article [7]. These formalizations are based on [8], [5], [3], and [4].

scholarly journals Effects of insulin on sperm cell quality in ram semen cooled at 5°C

2021 ◽  
Vol 42 (6supl2) ◽  
pp. 3803-3812
Author(s):  
Maurício Fraga van Tilburg ◽  
◽  
Rodrigo Vasconcelos de Oliveira ◽  
Carla Renata Figueiredo Gadelha ◽  
Bruno Fagundes ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Semen Analysis ◽  
Control Group ◽  
Computer Assisted ◽  
Insulin Group ◽  
Hypoosmotic Swelling Test ◽  
Semen Extender ◽  
Ram Sperm ◽  
Acrosomal Integrity

Insulin is present in the seminal plasma and is involved in sperm activities like motility and capacitation. However, the effects of insulin on the viability of cooled ram sperm are not fully understood. Therefore, the objective of the current study was to evaluate the effect of insulin addition on ram sperm maintained at 5ºC. Sperm samples were collected from six healthy, mature Santa Inês rams. The ejaculates were divided into two aliquots with (insulin group) or without (control group) insulin (3 IU mL-1) in the semen extender, and then cooled at 5°C for 48 hours. Subsequently, the sperm cells were evaluated for motility and kinetics using computer-assisted semen analysis. The samples were evaluated for acrosomal integrity by fluorescein using isothiocyanate combined with peanut agglutinin (FITC-PNA) and membrane functionality by the hypoosmotic swelling test. The semen analysis was performed after 24 or 48 hours of cooling. There was an increased percentage of progressive sperm motility (%), straightness (%), linearity (%) and beat caudal frequency (Hz) in the insulin group after 24 and 48 hours of cooling (p < 0.05). However, insulin did not affect total sperm motility, sperm velocities (VSL, VAP and VCL) (μm seg-1), acrosomal integrity and membrane functionality during cooling (p > 0.05). In conclusion, the addition of 3 IU mL-1 insulin to ram semen extender improved the quality of sperm motility after cooling.

Changes in sperm-bound amidase activity suggest subtle damage to ram sperm acrosomes by freezing/thawing, not detected by light microscopy

1996 ◽  
Vol 45 (1-2) ◽  
pp. 81-89 ◽  
Author(s):  
M.A. de las Heras ◽  
A. Valcarcel ◽  
C. Furnus ◽  
L. Pérez ◽  
D. Moses ◽  
...  
Keyword(s):  
Light Microscopy ◽  
Amidase Activity ◽  
Ram Sperm

Self-sacrifice for in-group's history: A diachronic perspective

2018 ◽  
Vol 41 ◽  
Author(s):  
Maria Babińska ◽  
Michal Bilewicz
Keyword(s):  
Clinical Psychology ◽  
Historical Events ◽  
Emotional Events ◽  
Reciprocal Process ◽  
The Way

AbstractThe problem of extended fusion and identification can be approached from a diachronic perspective. Based on our own research, as well as findings from the fields of social, political, and clinical psychology, we argue that the way contemporary emotional events shape local fusion is similar to the way in which historical experiences shape extended fusion. We propose a reciprocal process in which historical events shape contemporary identities, whereas contemporary identities shape interpretations of past traumas.

What is the purpose of cognition?

2020 ◽  
Vol 43 ◽  
Author(s):  
Aba Szollosi ◽  
Ben R. Newell
Keyword(s):  
Infinite Number ◽  
Human Cognition ◽  
The Way

Abstract The purpose of human cognition depends on the problem people try to solve. Defining the purpose is difficult, because people seem capable of representing problems in an infinite number of ways. The way in which the function of cognition develops needs to be central to our theories.

Peculiarity Parameters

1976 ◽  
Vol 32 ◽  
pp. 233-254
Author(s):  
H. M. Maitzen
Keyword(s):  
Magnetic Field ◽  
Magnetic Fields ◽  
Theoretical Work ◽  
Observational Evidence ◽  
Rotator Model ◽  
Peculiar Behaviour ◽  
Field Spectrum ◽  
Oblique Rotator ◽  
Ap Stars ◽  
The Way

Ap stars are peculiar in many aspects. During this century astronomers have been trying to collect data about these and have found a confusing variety of peculiar behaviour even from star to star that Struve stated in 1942 that at least we know that these phenomena are not supernatural. A real push to start deeper theoretical work on Ap stars was given by an additional observational evidence, namely the discovery of magnetic fields on these stars by Babcock (1947). This originated the concept that magnetic fields are the cause for spectroscopic and photometric peculiarities. Great leaps for the astronomical mankind were the Oblique Rotator model by Stibbs (1950) and Deutsch (1954), which by the way provided mathematical tools for the later handling pulsar geometries, anti the discovery of phase coincidence of the extrema of magnetic field, spectrum and photometric variations (e.g. Jarzebowski, 1960).

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