Biomedical subjects

A D Ealy

Publications and source records attributed to A D Ealy.

29 records · Page 2Linked to original sources

PubMed1998

Loss of the signature six carboxyl amino acid tail from ovine interferon-tau does not affect biological activity.

Interferon-tau (IFN-tau) is a type I IFN that is secreted from conceptuses of Bovidae (sheep, cattle, and related ruminant ungulates) for a few days during early pregnancy. It acts to prolong the life span of the corpus luteum. All secreted forms of IFN-tau, like the related IFN-omega, are 172 amino acids in length and differ from IFN-alpha and -beta by the presence of six additional amino acids at their carboxyl termini. The aim of this study was to determine whether this carboxyl tail was important for biological activity of IFN-tau, particularly for its antiluteolytic function in ewes. Full-length ovine IFN-tau (p3) and a mutated form truncated by six amino acids at its carboxyl terminal (p3Trn6, 166 amino acids) were produced in Escherichia coli. Both proteins had similar antiviral activities (2.12 +/- 0.92 x 10(8) IU/mg for p3; 1.96 +/- 0.58 x 10(8) IU/mg for p3Trn6) when tested on Madin-Darby bovine kidney (MDBK) cells. Antiproliferative activity, as measured on human Daudi cells by determining the protein concentration required to inhibit growth by 50%, was slightly higher (p < 0.05) for p3Trn6 (7.36 +/- 0.46 pM) than for p3 (13.99 +/- 0.85 pM). Most importantly, p3 and p3Trn6 were equally capable of prolonging the life span of the corpus luteum of nonpregnant ewes when the proteins were administered at doses of either 60 or 300 microg/day into the uterine lumen through indwelling uterine cannulae from Day 10 to Day 18 postestrus. Therefore, the carboxyl-terminal amino acid extension for IFN-tau does not appear to serve a functional role in the action of these proteins.

Animals↗

PubMed1997

Ontogeny of temperature-regulated heat shock protein 70 synthesis in preimplantation bovine embryos.

Development of the preimplantation embryo is very susceptible to disruption by heat shock. As embryos proceed through development, they acquire resistance to heat shock, perhaps because they become transcriptionally active and can respond to environmental changes by undergoing transcriptionally-regulated cellular adaptation. Objectives were to determine the ontogeny of heat shock protein 70 (HSP70) synthesis in preimplantation bovine embryos and to ascertain whether heat-induced increases in HSP70 in embryos are caused by environmental alterations in gene expression. Exposure of bovine embryos to heat shock induced synthesis of a 68 kDa form of HSP70 called HSP68 as early as the two-cell stage of development. Induction of HSP68 was alpha-amanitin independent at the two-cell stage but was blocked by alpha-amanitin as early as the early four-cell stage. Therefore, heat-induced synthesis of HSP68 is regulated at the level of transcription at a time before the major round of embryonic genome activation is considered to occur. Two other constitutive HSP70 molecules were identified called heat shock cognates (HSC) 71 and 70; both proteins were synthesized during all stages of development from the two-cell to hatched blastocyst stages. However, heat-induced synthesis of HSC71 and HSC70 was not evident until the expanded blastocyst stage. In conclusion, environmental signals can activate gene expression before the major round of embryonic genome activation occurs in bovine embryos. Moreover, differences in thermal sensitivity of early embryos to heat shock is not caused by an inability to synthesize HSP70, suggesting that other mechanisms are involved in developmental acquisition of thermotolerance.

Amanitins↗

PubMed1995

Regulation of heat shock protein 70 synthesis by heat shock in the preimplantation murine embryo.

Induced thermotolerance in murine embryos occurs at the 8-cell stage when embryos are maintained in vitro but not until the blastocyst stage if development proceeds in vivo. Present results indicate that ability of embryos to undergo induced thermotolerance is not limited by heat shock protein 70 (HSP70) synthesis. Exposure of 8-cell embryos to 40 degrees C enhanced synthesis of 2 constitutive HSP70 proteins (HSC70 and HSC72) and induced another protein, HSP68; exposure of 43 degrees C was required to induce similar responses in expanded blastocysts. Unlike induced thermotolerance, increased synthesis of HSP70 molecules did not depend on whether embryos were cultured or developed in vivo. Thus, other biochemical mechanisms in addition to HSP70 confer thermotolerance in the preimplantation-stage murine embryo. The observation that the temperature threshold for induction of HSP70 synthesis increased from the 8-cell to the blastocyst stage is indicative of these other biochemical processes.

Journal Article↗

PubMed1995

Effect of heat shock on function of frozen/thawed bull spermatozoa.

Deposition of spermatozoa in the reproductive tract of hyperthermic cows could conceivably result in sperm damage. Accordingly, a series of experiments tested the effects of heat shock on functional characteristics and free radical production of bull spermatozoa. Viability was reduced slightly by short-term (1 to 3 h) culture at 42 and 43 degrees C as compared with culture at 39 degrees C. There was no effect of culture at 42 degrees C on the ability of spermatozoa to undergo swim-up or of 42 degrees C on the percentage of motile spermatozoa. However, exposure to 41 degrees C for 3 h reduced percentage of motile sperm, 41 and 42 degrees C reduced sperm velocity and 43 degrees C decreased the proportion of spermatozoa undergoing swim-up. In other experiments, there was no effect of heat shock (41 or 42 degrees C for 1 to 3 h) on DNA integrity, presence of intact acrosomes, or fertilizing ability of the spermatozoa. Superoxide production by spermatozoa was higher at 42 degrees C than at 39 or 41 degrees C, but there was no detectable hydrogen peroxide production at any temperature. The antioxidant, glutathione, tended to improve the ability of spermatozoa to undergo swim-up at 39 degrees C but not at 43 degrees C. Taken together, these results suggest that heat shock of a magnitude similar to that seen in vivo (41 to 42 degrees C) has little effect on sperm functions that affect fertilizing capability.

Journal Article↗

PubMed1995

Evidence that glutathione is involved in thermotolerance of preimplantation murine embryos.

Experiments were conducted to determine whether or not glutathione (GSH) is involved in thermotolerance responses of murine morulae. In the first experiment, morulae were exposed to either homeothermic temperature (37 degrees C), mild heat shock (40 degrees C for 1 h), severe heat shock (43 degrees C for 2 h), or a mild heat shock followed by severe heat shock (to induce thermotolerance). Exposure to mild heat shock did not affect viability and development, but severe heat shock reduced viability (i.e., live/dead staining) and the proportion of morulae that developed to blastocysts. This effect of 43 degrees C was reduced if embryos were first exposed to a mild heat shock of 40 degrees C. In the presence of DL-buthionine-[S,R]-sulfoximine (BSO), an inhibitor of GSH synthesis, the ability of 40 degrees C to confer thermotolerance was reduced. BSO decreased embryonic GSH content but did not decrease overall protein synthesis. In another experiment, administration of S-adenosyl-L-methionine, an inducer of GSH synthesis, decreased the deleterious effects of heat shock of 43 degrees C for 2 h on viability and percentage of embryos that became blastocysts. Addition of 5 microM GSH or GSH ester reduced the effect of 42 degrees C for 2 h on viability but not on continued development. The results suggest a role for GSH-dependent mechanisms in the processes by which murine embryos limit deleterious effects of heat shock.

Animals↗

PubMed1995

Developmental changes in sensitivity of bovine embryos to heat shock and use of antioxidants as thermoprotectants.

Experiments were conducted with in vitro-produced bovine embryos to determine whether 1) increased culture temperatures (i.e., heat shock) adversely affected embryonic development, 2) embryos become more resistant to heat shock as they advance in development, and 3) selective antioxidant molecules alleviate heat shock effects on embryonic development. Development of 2-cell embryos to > or = 16-cell stage on d 5 after in vitro fertilization was not affected by a heat shock of 40 degrees C for 3 h, but 41 or 42 degrees C for 3 h decreased (P = .004) development. In a separate experiment, development of 2-cell embryos was decreased (P = .01) by exposure to 41 degrees C for 3 h but not for 1 h. In contrast, development of morulae to blastocysts was not affected by heat shock of 41 degrees C for 1 or 3 h. Medium supplementation with 50 nM glutathione or 50 mM taurine before heat shock did not reduce the effects of heat shock (41 degrees C for 3 h) on 2-cell embryos. Likewise, addition of glutathione ester, a more membrane-permeable analog of glutathione, did not protect 2-cell embryos from heat shock. In conclusion, early bovine embryos are susceptible to disruption in development caused by heat shock. As embryos progress in development, they acquire resistance to heat shock. Glutathione, taurine, and glutathione ester were not effective in alleviating the effects of heat shock on development of 2-cell embryos. Consequently, molecules have yet to be identified that can protect early-stage bovine embryos from the adverse effects of heat shock.

Animals↗

PubMed1994

Induced thermotolerance during early development of murine and bovine embryos.

During early development, elevated temperatures have deleterious effects on embryonic viability and development. The primary objective of the current study was to determine the ontogeny of induced thermotolerance during early murine embryonic development. Embryos were either retrieved from superovulated ICR female mice at the 2 cell and 4 cell stages and cultured thereafter or were retrieved from oviducts or uterine horns at the desired stage of development. Induction of thermotolerance was detected by evaluating viability and further development after embryos were exposed to homeothermic temperature (37 degrees C), mild heat shock (40 degrees C for 1 h), severe heat shock (42 degrees C for 1 h or 43 degrees C for 2 h), or mild heat shock followed by severe heat shock (to induce thermotolerance). Induction of thermotolerance was observed beginning at the 8 cell stage when embryos were developed in culture from the 2 cell to 4 cell stage. When embryos were developed in vivo (i.e., were retrieved from the reproductive tract at the desired stage of development), thermotolerance was not induced until the blastocyst stage of development. The induction of thermotolerance was dependent on serum supplementation since induction of thermotolerance was not observed when embryos were placed in medium without serum. Induced thermotolerance could also be demonstrated in bovine blastocysts. In conclusion, embryos acquire the ability to undergo thermotolerance as they progress through development. The timing of processes leading to acquisition of thermotolerance can, however, be hastened by exposure of embryos to in vitro conditions.

Adaptation, Physiological↗

PubMed1994

Effectiveness of short-term cooling and vitamin E for alleviation of infertility induced by heat stress in dairy cows.

Four experiments were performed to determine whether cooling cows during final maturation of oocytes and early embryonic development or injection of vitamin E at AI prevented adverse effects of heat stress on pregnancy rates in lactating Holstein dairy cows. In Experiment 1, cows were placed in a cooling facility containing sprinklers and forced ventilation or received shade only from 2 to 3 d before until 5 to 6 d after breeding. Although cooling had no effect on detection of estrus, pregnancy rates were increased slightly for cooled cows (8 of 50 cows; 16.0%) compared with those for cows exposed to shade only (2 of 32 cows; 6.2%). In Experiments 2 through 4, cows were administered 3000 IU of vitamin E or placebo i.m. at AI during two consecutive summers and one winter in Florida. Administration of vitamin E had no consistent beneficial effect on pregnancy rates during summer or winter. Short-term cooling improved pregnancy rates slightly in heat-stressed cows, but administration of vitamin E had no beneficial effects on pregnancy rates during heat stress. Further improvements in cooling schemes during early pregnancy and delineation of antioxidant effects are necessary before such systems become practical for improvement of fertility in heat-stressed dairy cows.

Animals↗

PubMed1993

Developmental changes in embryonic resistance to adverse effects of maternal heat stress in cows.

The objective of this study was to determine whether bovine embryos become more resistant to deleterious effects of maternal heat stress as early embryonic development progresses. Superovulated, lactating Holstein cows were bred by AI and assigned to be heat stressed on d 1, 3, 5, or 7 of pregnancy (d 0 = day of estrus) or not heat stressed (control). Embryos were retrieved from the uterus on d 8 and evaluated for viability and stage of development. Compared with embryos of control cows, embryos of cows receiving heat stress on d 1 had decreased viability and development. Maternal heat stress on other days had no detrimental effect on embryonic viability or stage of development. Bovine embryos become more resistant to adverse effects of maternal heat stress as pregnancy progresses; substantial resistance develops by d 3. This information may be useful in design of environmental modification systems that provide cooling at critical periods of gestation to enhance pregnancy rates during summer in hot climates.

Animals↗

PubMed1992

Thermoprotection of preimplantation bovine embryos from heat shock by glutathione and taurine.

To determine if deleterious effects of heat shock on embryos could be reduced in vitro by glutathione or taurine, morulae from superovulated cows were placed in modified Hams-F10 medium supplemented with 50 nM glutathione (GSH), 50 mM taurine or neither. Morulae were incubated for 2 hours at 38.5 degrees C, then at 42.0 degrees C (heat shock) or 38.5 degrees C for 2 hours and followed by incubation at 38.5 degrees C for 20 hours. Neither GSH nor taurine enhanced viability or blastocyst development at 38.5 degrees C. At 42.0 degrees C, however, GSH and taurine increased (P less than 0.02) viability (73%, 41% and 26% live for GSH, taurine and control); GSH increased (P less than 0.05) blastocyst development (55% for GSH vs. 30% for control). In conclusion, partial thermoprotection of bovine embryos from heat shock can be achieved in vitro by administration of GSH. Taurine is only slightly effective.

Animals↗

PubMed1990

Energy balance and body condition influence luteal function in Holstein heifers.

A factorial experiment was conducted to determine influence of energy balance (EB) and body condition (BC) on luteal function in heifers. Heifers with moderate (MBC) or fat (FBC) BC were fed individually to sustain positive EB (PEB) or to cause negative EB (NEB). Intake of feed was measured daily and body weight weekly. Progesterone was quantified daily in serum for 3.5 estrous cycles. On days 9, 10, or 11 after fourth estrus, blood was sampled every 15 min for 12 hr to quantify luteinizing hormone (LH), growth hormone (GH), insulin and non-esterified fatty acids (NEFA). The next day, luteal cells were incubated and proportions of small to large cells were determined. After fourth estrus, area of progesterone profiles in serum for 10 days postestrus was reduced in all heifers relative to MBC-PEB heifers. But, luteal weight from FBC-PEB and MBC-NEB heifers was less than MBC-PEB heifers and FBC-NEB heifers were intermediate. Secretion of progesterone in vitro was increased by LH for PEB but not NEB heifers. MBC-NEB heifers had increased ratios of small to large luteal cells. Independent of BC, NEB decreased concentrations of insulin and increased GH and NEFA. Secretion of progesterone was not associated with LH, GH or insulin, but was correlated negatively with NEFA. We conclude that reduced concentrations of progesterone in serum of FBC-PEB and MBC-NEB heifers is due to impaired luteal development. But, reduced concentrations of progesterone in serum of NEB heifers is due also to reduced basal (MBC) and LH-induced (MBC and FBC) secretion of progesterone by luteal cells. Body condition at onset of NEB may determine when effects of NEB on progesterone are detected.

Animals↗