The Effect of Temperature on the Survival In Vitro of Rabbit Spermatozoa Obtained from the Vagina

1930 ◽  
Vol 7 (2) ◽  
pp. 175-195
Author(s):  
J. HAMMOND

1. The time during which spermatozoa, obtained from the vagina of the female rabbit after service, retain their fertilising power when kept in vitro at diferent temperatures has been determined. 2. At 35° C., or just below body temperature, young are obtained up to 14 hours at 10° C. up to 96 hours and at 0° C. up to 16 hours, there being a well-marked optimum temperature for survival. 3. Not only does the percentage of animals fertile fall with increase in the length of time the semen is kept, but also the average size of the litter decreases in the small litters so obtained the birth weight is increased considerably and the duration of pregnancy slightly lengthened. 4. The percentage of spermatozoa motile, when examined at room temperature is a fairly good indication of the percentage fertility. 5. The sex-ratio is not materially affected by keeping the sperm. 6. Sterility due to adherent tubes was frequent during the course of the experiments. 7. These results with semen obtained from the vagina are compared with those obtained in a parallel series direct from the epididymis of the male under paraffin

1930 ◽  
Vol 7 (2) ◽  
pp. 201-219
Author(s):  
ARTHUR WALTON

1. A technique is described for the investigation of in vitro problems with spermatozoa removed from the vas deferens of the rabbit. The method involves the protection of the spermatozoa by means of liquid paraffin from evaporation of rapid gaseous exchange. 2. The survival of functional integrity (fertilizing capacity) has been shown to be a function of temperature and the effect of temperature has been studied over the range from 45° C. to 0° C. Above body temperature the spermatozoa are rapidly destroyed. At body temperature (40° C.) maximal survival is about 13 hours. As the temperature is lowered survival becomes increasingly prolonged until a maximum of 7 days is reached at 15°C. The curve over the range from 15° C. to 40° C. is only approximately exponential and it is doubtful whether an analogy can be drawn between the effect of temperature on the velocity with which the spermatozoa are destroyed, and the effect of temperature on the velocity of many biological reactions which follow approximately the van't Hoff and Arrhenius questions. Below the optimum temperature (10°-15°C.) the velocity of destruction is acclerated by fall of temperature. 3. The sex-ratio of the resulting offspring is not significantly altered by keeping the spermatozoa outside the body.


1973 ◽  
Vol 29 (01) ◽  
pp. 183-189
Author(s):  
C. A Praga ◽  
E. M Pogliani

SummaryTemperature represents a very important variable in ADP-induced platelet aggregation.When low doses of ADP ( < 1 (μM) are used to induce platelet aggregation, the length of the incubation period of PRP in the cuvette holder of the aggregometer, thermostatted at 37° C, is very critical. Samples of the same PRP previously kept at room temperature, were incubated for increasing periods of time in the cuvette of the aggregometer before adding ADP, and a significant decrease of aggregation, proportional to the length of incubation, was observed. Stirring of the PRP during the incubation period made these changes more evident.To measure the exact temperature of the PRP during incubation in the aggre- gometer, a thermocouple device was used. While the temperature of the cuvette holder was stable at 37° C, the PRP temperature itself increased exponentially, taking about ten minutes from the beginning of the incubation to reach the value of 37° C. The above results have a practical significance in the reproducibility of the platelet aggregation test in vitro and acquire particular value when the effect of inhibitors of ADP induced platelet aggregation is studied.Experiments carried out with three anti-aggregating agents (acetyl salicyclic acid, dipyridamole and metergoline) have shown that the incubation conditions which influence both the effect of the drugs on platelets and the ADP breakdown in plasma must be strictly controlled.


2012 ◽  
Vol 06 (04) ◽  
pp. 361-369 ◽  
Author(s):  
Serdar Baglar ◽  
Adil Nalcaci ◽  
Mustafa Tastekin

ABSTRACTObjective: The aim of this study was to examine the effect of temperature on fluoride uptake by enamel specimens from a 0.05% NaF-fluoridated mouthrinse (Oral-B Advantage; Oral-B Laboratories, Newbridge, UK).Methods: Enamel specimens were prepared from extracted human maxillary central incisors. A fluoride-specific ion electrode was used to measure the uptake from a 2 ppm fluoride solution containing 50.0 mL of distilled water, total ion strength adjustment buffer, and fluoridated rinse at 3 different temperatures (room temperature, 25°C; human body temperature, 37°C; hyper-fever temperature, 43°C). One-way analysis of variance and least significant difference were used to assess intragroup and intergroup differences (P<.05).Results: The study found that both the amount and the rate of fluoride uptake increased significantly with increase in temperature. This effect was particularly noticeable at 43°C.Conclusions: The temperature of the NaF mouthrinse may easily and safely be increased beyond room temperature by placing a container of the NaF mouthrinse in a bowl of hot water, allowing greater fluoride penetration into the enamel from the mouthrinse when used at home as a routine prophylactic agent. (Eur J Dent 2012;6:361-369)


1988 ◽  
Vol 140 (1) ◽  
pp. 421-435 ◽  
Author(s):  
JON M. HARRISON

In this study, I examine the effect of temperature on haemolymph acid-base status in vivo and in vitro in the two-striped grasshopper Melanoplus bivittatus. Melanoplus bivittatus experience wide (up to 40 °C) diurnal body temperature fluctuations in the field, but maintain body temperature relatively constant during sunny days by behavioural thermoregulation. Haemolymph pH was statistically constant (7.12) between 10 and 25°C, but decreased by −0.017 units °C− from 25 to 40°C. Relative alkalinity and fractional protein dissociation were conserved only at body temperatures at which feeding and locomotory activity occur, above 20°C. Haemolymph total CO2 (Ctot) increased from 10 to 20°C and decreased from 20 to 40°C. Haemolymph Pco2 increased from 10 to 20°C and was statistically constant between 20 and 40°C. Carbonic acid pKapp in haemolymph was 6.122 at 35°C, and decreased with temperature by −0.0081 units°C−1. Haemolymph buffer value averaged −35mequivl−1pHunit−1. Haemolymph pH changes with temperature were small (less than −0.004 units°C−1) in vitro at constant Pco2. Therefore, passive physicochemical effects cannot account for the pattern of acid-base regulation in vivo. The temperature shift from 10 to 20°C was accompanied by a net addition of 4.2-6.2 mmoll−1 of bicarbonate equivalents to the haemolymph. The temperature shift from 20 to 40°C was accompanied by a net removal of 10–14 mmoll−1 of bicarbonate equivalents from the haemolymph. Haemolymph acid-base regulation in vivo during temperature changes is dominated by active variation of bicarbonate equivalents rather than by changes in Pco2 as observed for most other air-breathers.


2011 ◽  
Vol 19 (9) ◽  
pp. 781-788 ◽  
Author(s):  
Zhi Huang ◽  
Bo Yu ◽  
Qingling Feng ◽  
Songjian Li

The chitosan/β-glycerophosphate (C/GP) system, which allows injection at room temperature and gel-formation at body temperature, can be considered as a candidate for injectable scaffolds. To improve the cytocompatiblity of the C/GPhydrogel, we used NaHCO3 as buffer to neutralize chitosan solution and lower the GP concentration from 400 to 135 mM. The chitosan/β-glycerophosphate//NaHCO3 (C/GP/NaHCO3) system was liquid before injection and became a three-dimensional porous hydrogel at body temperature, as revealed by scanning electron microscopy (SEM). The C/GP/NaHCO3 hydrogels showed higher cytocompatibility with rat bone marrow-derived mesenchymal stem cells (rBMSCs) in vitro compared to C/GP hydrogels. Our results indicated that the C/GP/NaHCO3 system was suitable as an injectable scaffold for tissue engineering.


2005 ◽  
Vol 17 (2) ◽  
pp. 270 ◽  
Author(s):  
L.S.A. Camargo ◽  
J.H.M. Viana ◽  
A.A. Ramos ◽  
W.F. de Sa ◽  
A.M. Ferreira ◽  
...  

Gir cattle are a popular zebu dairy breed in tropical and subtropical regions because of their tolerance of heat stress and resistance to tick-borne disease. The use of in vitro embryo production (IVP) technology may help accelerate genetic improvement of this breed. However, in general, IVP systems have been implicated in the production of large offspring and a greater proportion of male calves. Natural breeding results in newborn Gir calves weighing around 25 kg despite the fact that dams may weigh over 500 kg. It is unknown whether in vitro-produced Gir embryos also result in large offspring. The aim of this work was to evaluate the effect of in vitro embryo production on gestation length, birth weight, and sex ratio in Gir cattle. COCs were harvested by oocyte pickup from mature non-lactating Gir cows and in vitro-matured in TCM 199 medium (Gibco, Sao Paulo, Brazil) with 10% inactivated estrous cow serum for 24 h under 5% CO2 at 38.5°C in air. Gir spermatozoa were obtained through the swim-up method and co-incubated with oocytes in Fert-TALP media (Parrish JJ et al. 1988 Biol. Reprod. 38, 1171–1180) with 10 μg/mL heparin (Sigma, Sao Paulo, Brazil) and 6 mg/mL fatty acid-free bovine albumin (Sigma) for 18 h in 5% CO2 at 38.5°C in air. Presumptive zygotes were co-cultured with their own cumulus cells in CR2aa medium (Wilkinson RF et al. 1996 Theriogenology 45, 41–49) with 10% fetal calf serum in humid atmosphere of 5% CO2 at 38.5°C in air. Fresh Day 7 blastocysts were transferred to synchronized B. indicus × B. taurus crossbred recipients. Data of gestation length, birth weight, and gender ratio from 26 IVP calves (IVP group) were recorded and compared to data obtained from Gir calves produced by artificial insemination or natural mating (n = 24; control group) using ANOVA or chi-square analysis. There was no difference (P > 0.05) in gestation length between pregnancies of the IVP and control groups (mean ± SEM, 285.4 ± 1.5 vs. 284.4 ± 1.1 days, respectively). IVP and control calves showed similar (P > 0.05) weight at calving (29.6 ± 0.9 vs. 26.9 ± 1.2 kg for IVP and control male calves, and 27.0 ± 2.5 vs. 25.2 ± 0.5 kg for IVP and control female calves, respectively). The percentage of male calves was greater (P < 0.05) in the IVP group than in the control group (76.9% vs. 43.4%, respectively). IVP calves did not show abnormalities associated with Large Offspring Syndrome, such as breathing difficulty and perinatal death. These data suggest that in vitro production may affect the development of Gir embryos, biasing the sex ratio in a manner similar to previously reported for in vitro-produced embryos from Bos taurus breeds. This work was supported by FAPEMIG and CNPq.


1988 ◽  
Vol 137 (1) ◽  
pp. 529-548 ◽  
Author(s):  
J. N. Stinner ◽  
R. L. Wardle

The effect of temperature upon respiratory exchange ratio (R) was measured in snakes (Coluber constrictor) and turtles (Chrysemys scripta). Increasing body temperature produced a transient elevation of R, and lowering body temperature transiently depressed R. These thermal effects resulted from an ‘excess’ and a ‘deficit’ CO2 elimination, respectively. Steady-state blood CO2 content (CCO2) in C. constrictor decreased linearly with rising temperature. Plasma bicarbonate concentration, calculated from in vivo arterial PCO2 and pH, followed the same pattern. Also, time courses of blood CCO2 were consistent with the metabolic studies. Less than half of the change in blood CCO2 could be explained by shifts of the in vitro CO2 dissociation curve; the remainder was contributed by other tissues. Blood lactate levels changed little with temperature. Based upon the blood studies, the predicted quantity of CO2 eliminated from the extracellular space when temperature increases is about 29% of the excess CO2 eliminated from the snakes. Thus, CCO2 in other tissues also decreases with rising temperature. It is concluded that reptiles function as open systems with respect to CCO2, which does not agree with alphastat control. Systemic arterial PO2 and PCO2 increased with rising body temperature in C. constrictor. The mechanisms producing these increases are discussed.


2021 ◽  
Vol 21 (8) ◽  
pp. 4151-4156
Author(s):  
Jeong Jun Kim ◽  
Jong Kook Lee

Zirconia ceramics has a bioinert property with low bioactivity. So, it is necessary to improve its low bioactivity by the surface modification using effective coating methods. In this study, we fabricated the hydroxyapatite-coated zirconia substrate by room temperature spray processing to improve the bioactivity of the zirconia implant and investigated its coating effect on the biological performance of zirconia substrate via an in vitro test in simulated body fluid (SBF) solution. Before the room temperature spray coating was completed, size-controlled hydroxyapatite powder that had an average size of 4.5 μm, was obtained by the calcination and milling of a commercial powder. By controlling the processing parameters, such as spraying distance, and deposition cycles, we fabricated homogeneous and dense hydroxyapatite coatings on zirconia substrate. Surface morphology, coating thickness, and microstructure were dependent on deposition cycles, and were related to surface roughness and bioactivity. Zirconia substrates with wave-patterned and roughened hydroxyapatite coatings demonstrated high bioactivity in their in vitro tests. Via the immersion test in an SBF solution, surface dissolution and new precipitates of hydroxyapatite were observed on coated zirconia substrate, indicating the degree of bioactivity.


Author(s):  
C. Jennermann ◽  
S. A. Kliewer ◽  
D. C. Morris

Peroxisome proliferator-activated receptor gamma (PPARg) is a member of the nuclear hormone receptor superfamily and has been shown in vitro to regulate genes involved in lipid metabolism and adipocyte differentiation. By Northern analysis, we and other researchers have shown that expression of this receptor predominates in adipose tissue in adult mice, and appears first in whole-embryo mRNA at 13.5 days postconception. In situ hybridization was used to find out in which developing tissues PPARg is specifically expressed.Digoxigenin-labeled riboprobes were generated using the Genius™ 4 RNA Labeling Kit from Boehringer Mannheim. Full length PPAR gamma, obtained by PCR from mouse liver cDNA, was inserted into pBluescript SK and used as template for the transcription reaction. Probes of average size 200 base pairs were made by partial alkaline hydrolysis of the full length transcripts. The in situ hybridization assays were performed as described previously with some modifications. Frozen sections (10 μm thick) of day 18 mouse embryos were cut, fixed with 4% paraformaldehyde and acetylated with 0.25% acetic anhydride in 1.0M triethanolamine buffer. The sections were incubated for 2 hours at room temperature in pre-hybridization buffer, and were then hybridized with a probe concentration of 200μg per ml at 70° C, overnight in a humidified chamber. Following stringent washes in SSC buffers, the immunological detection steps were performed at room temperature. The alkaline phosphatase labeled, anti-digoxigenin antibody and detection buffers were purchased from Boehringer Mannheim. The sections were treated with a blocking buffer for one hour and incubated with antibody solution at a 1:5000 dilution for 2 hours, both at room temperature. Colored precipitate was formed by exposure to the alkaline phosphatase substrate nitrobluetetrazoliumchloride/ bromo-chloroindlylphosphate.


1990 ◽  
Vol 80 (3) ◽  
pp. 431-436 ◽  
Author(s):  
Isabelle Delvallee ◽  
Annie Paffen ◽  
Geert-Jan De Klerk

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