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Biomedical subjects

R A Mead

Publications and source records attributed to R A Mead.

At least 37 records · Page 2Linked to original sources

Psychological differentiation, arousal, and lipreading efficiency in hearing-impaired and normal children.

Psychological differentiation, psychophysiological arousal, hearing capacity, and ability to lipread unrelated sentences presented in a silent technicolor film were investigated in 62 children (mean age 10 years, 4 months), classified into three hearing categories (normal, mild to moderate hearing loss, severe hearing loss) based on their average hearing threshold in the range of speech frequencies for their best ear. All subjects were administered the Rod and Frame Test, Craig Lipreading Inventory, and skin conductance measures under two conditions (pre-task and task). The results showed that the ability to lipread sentences from a film was related significantly to psychological differentiation (cognitive style) and pre-task conductance level, but was not related to hearing capacity or task conductance level. The best predictors of lipreading ability were successful performance on the Rod and Frame Test and high increases in skin conductance from pre-task to task situation.

Arousal↗

Role of prolactin and luteinizing hormone in regulating timing of implantation in the spotted skunk.

The western spotted skunk exhibits an obligate delay of implantation lasting 200-220 days. The pituitary is essential for luteal activation. The corpora lutea, in turn, secrete the hormones necessary for blastocyst implantation. Two experiments were designed to determine which pituitary hormones are responsible for increasing luteal activity and induction of implantation. Forty-two pregnant skunks with delayed implanting blastocysts were treated as follows: 13 served as untreated controls, 6 received 0.5 mg prolactin (PRL) daily, and 5 received diluent beginning in January. Four received 1.5 mg bromocriptine (CB-154) daily, 3 received both CB-154 and PRL, 3 received diluent, 5 received a gonadotropin-releasing hormone agonist (GnRHa) dispensed from osmotic minipumps, and 3 received diluent dispensed from osmotic minipumps starting in April. The skunks were subjected to a natural photoperiod. Duration of preimplantation and blood levels of progesterone and luteinizing hormone were measured. PRL significantly (p less than 0.05) shortened and CB-154 significantly (p less than 0.05) prolonged the duration of preimplantation when compared to controls (148 +/- 33.6 vs. 251 +/- 3.2 vs. 199 +/- 5.1 days, respectively). PRL was able to reverse the inhibitory effect of CB-154 when both were administered simultaneously (195 +/- 4.0 vs. 251 +/- 3.2 days). GnRHa had no significant (p greater than 0.05) effect on duration of preimplantation (199 +/- 5.1 days) when compared to controls (203 +/- 3.2 days). These results indicate that PRL is the primary pituitary hormone responsible for increased luteal activity and subsequent blastocyst implantation in the spotted skunk.

Animals↗

Lesions to the anterior hypothalamus prevent the melatonin-induced lengthening of delayed implantation.

The anterior hypothalamic area (AHA) has been postulated as a site of action for melatonin. We tested the hypothesis that lesions to the AHA (AHAx) would counteract the inhibitory effect of exogenous melatonin on blastocyst implantation in the spotted skunk by removing a possible site of action. Forty-seven females were treated as follows during delayed implantation. In Exp 1, five received empty Silastic capsules, five received Silastic capsules containing melatonin, six received sham AHAx plus empty capsules, none received AHAx plus empty capsules, and eight received AHAx plus capsules containing melatonin. In Exp 2, four skunks each received two empty capsules, five skunks each received two capsules containing melatonin, and five skunks received AHAx plus capsules containing melatonin. All capsules were inserted sc in the interscapular region 14-35 days after surgery in Exp 1 and 2 weeks before surgery in Exp 2. Surgery was performed between January 22 and February 12, 1988, in Exp 1 and on March 2-3, 1989, in Exp 2. The skunks were subjected to a natural photoperiod, and the duration of preimplantation was measured. In Exp 1, AHAx plus empty capsules significantly (P less than 0.05) shortened the duration of preimplantation (163 +/- 14.7 days) compared to that in sham AHAx or intact controls (193 +/- 26.1 and 188 +/- 10.6 days, respectively). Melatonin significantly (P less than 0.05) prolonged the duration of preimplantation (289 +/- 2.9 days) in intact skunks, but failed to do so in skunks with AHAx, as the preimplantation period was significantly shortened (159 +/- 6.1 days). In Exp 2, AHAx reversed the inhibitory effect of melatonin on the duration of preimplantation (191 +/- 21.5 days), as intact melatonin-treated skunks had a significantly longer preimplantation period (260 +/- 2.5 days) than skunks receiving empty capsules (191 +/- 16.4 days). The inhibitory effect of melatonin was reversible in all intact skunks, as blastocysts implanted 23 days, on the average, after cessation of treatment with melatonin. These data are consistent with the hypothesis that a portion of the AHA and/or adjacent regions play an essential role in timing blastocyst implantation in the spotted skunk. The lesions may have given this result by ablating a neural pathway controlling PRL secretion and may or may not have involved a site of action for melatonin.

Animals↗

Hormonal induction of oestrus and pregnancy in anoestrous ferrets (Mustela putorius furo).

Three doses of FSH were tested for their ability to induce oestrus in ferrets. A dose of 0.25 mg, administered twice daily, induced oestrus and breeding in most females within 6-13 days. Addition of 5 i.u. hCG during the final stages of follicular development enhanced the percentage of females with implantation sites (85%), and 23% of the ferrets so treated gave birth to kits, none of which survived for more than 3 days. This may have been due to insufficient prolactin secretion and thus inadequate luteal maintenance and milk production, as these females were maintained on a short-day photoperiod.

Anestrus↗

Studies on the mechanism of action of prostaglandin F2 alpha induced luteolysis in rats.

The effects of prostaglandin F2 alpha (PGF2 alpha) administration on the utilization of low density lipoprotein (LDL) and progesterone secretion were examined in dispersed luteal cells from rat ovaries. Immature rats were rendered pseudopregnant with administration of pregnant mare serum gonadotropin and human chorionic gonadotropin. Animals were sacrificed at different times after PGF2 alpha (5 mg/kg) or vehicle administration on day-5 of pseudopregnancy. Administration of PGF2 alpha in vivo decreased human chorionic gonadotropin (hCG) binding to luteal cell membranes in vitro but enhanced binding of LDL. Utilization of labelled cholesterol for steroid synthesis from reconstituted LDL [(3H)-CL-LDL] by dispersed luteal cells was enhanced following PGF2 alpha administration. This suggests that the LDL pathway is not suppressed during prostaglandin induced luteolysis. Progesterone and total progestin secretion in response to N6-2'-0-Dibutyryladenosine 3'5'-cyclic monophosphate (cAMP) was decreased at 2, 4 and 24 hours following PGF2 alpha administration demonstrating a post-cAMP defect in steroidogenesis. Addition of the hydroxylated sterols, 20 or 25-OH cholesterol as substrate stimulated progesterone secretion in vehicle treated rats in a dose dependent fashion with 20-OH cholesterol being more potent. Progesterone secretion in response to stimulation with luteinizing hormone (LH) and cAMP from vehicle treated rats was less than that observed with 20 or 25-OH cholesterol, indicating that endogenous substrate may be a limiting factor in steroid synthesis. The maximal capacity of luteal tissue to produce progestins following PGF2 alpha administration was determined with 20-OH cholesterol as the substrate. The results suggest that the post-cAMP defect at 4 hours following PGF2 alpha administration may be due to failure of the cells to mobilize endogenous cholesterol. However at 24 hours following PGF2 alpha administration the decreased ability of luteal cells to convert cholesterol to pregnenolone may contribute to decreased progesterone synthesis.

Animals↗

Partial characterization of a luteal factor that induces implantation in the ferret.

This study was designed to test the hypothesis that ferret corpora lutea (CL) secrete a compound that acts in conjunction with progesterone to induce blastocyst implantation and to identify the chemical nature of this compound. CL and the residual ovarian tissue, obtained predominantly on the ninth day of pseudopregnancy, were extracted with 0.05 M phosphate-buffered saline. The extracts were injected into pregnant ferrets that had been ovariectomized on Day 6 of pregnancy and had received Silastic implants containing progesterone. Aqueous luteal extracts, but not those of the residual ovarian tissue, induced implantation in test animals. Fractionation of the luteal extracts by passage through a series of filters with molecular weight (MW) cutoffs ranging from 500 to 50,000 consistently revealed that the biologically active fraction was retained on the filter with the highest MW cutoff employed. Moreover, blastocyst implantation failed to occur in ovariectomized, progesterone-treated ferrets after one-half of a luteal preparation (MW greater than 50,000) was incubated with a broad-spectrum protease. These data are consistent with the hypothesis that CL of the ferret secrete a protein during the preimplantation period that is essential for blastocyst implantation.

Animals↗

Size distribution of ferret luteal cells during pregnancy.

Steroidogenic cells in the corpus luteum of the ferret (Mustela putorius) during early (Days 6 and 13) to midpregnancy (Day 24) were characterized using electron microscopy, immunocytochemical localization of neurophysin, and smears of dispersed cells obtained by dissociating luteal cells with collagenase. The latter were stained for 3 beta-hydroxysteroid dehydrogenase (3 beta-HSD) activity, and the diameters of the cells were determined with an ocular micrometer. Very small cells (less than 12 microns) stained negative for 3 beta-HSD, occurred in clumps of 5-50 cells, and were presumed to be primarily endothelial cells. 3 beta-HSD-positive cells covered a wide spectrum of sizes ranging from 14 to 56 microns and did not exist as two discrete populations. The ratio of small (less than 25 microns) to large (greater than 25 microns) cells was 1.86:1.0 on Day 6, with the 17- to 20-microns cell size class predominating. On the day of implantation (Day 13), about 75% of the cells ranged from 26 to 50 microns, with the 29-microns size predominating. By Day 24, the ratio of small-to-large cells had declined to 0.15. Nearly 90% of the cells were in the 26- to 56-microns range, the predominant size being 35 microns. All size classes of luteal cells stained negative for neurophysin on all 3 days of pregnancy studied. Luteal cells obtained on Days 6, 13, and 24 of pregnancy failed to reveal any evidence of mitosis after in vivo or in vitro colchicine treatment. We interpret these results as indicating that the 3 beta-HSD-positive luteal cells of ferrets progressively increase in size as small luteal cells complete their differentiation from granulosa cells and ultimately form larger luteal cells with somewhat different ultrastructural characteristics.

Animals↗

Effects of suprachiasmatic nuclear ablation and melatonin on delayed implantation in the spotted skunk.

The suprachiasmatic nuclei (SCN) have been implicated as neural timers of reproductive events and as possible sites of action for melatonin. We tested the hypothesis that ablation of the SCN (SCNx) would counteract the inhibitory effect of exogenous melatonin in blastocyst implantation in the spotted skunk by removing a possible site of action. Thirty-eight pregnant females with unimplanted blastocysts were treated as follows: 4 served as untreated controls, 6 received empty Silastic capsules, 5 received Silastic capsules containing melatonin, 10 received sham lesions in the SCN, 7 received lesions in the SCN and Silastic capsules containing melatonin, and 6 received lesions in the SCN and empty Silastic capsules. All surgical treatments were completed by February 15. The skunks were subjected to a natural photoperiod, and the duration of preimplantation was measured. The lesions destroyed an average of 87 +/- 10% of the SCN and a small amount of the surrounding hypothalamus. SCNx had no significant effect on duration of preimplantation (200 +/- 25.6 days) when compared to sham SCNx (205 +/- 21.5 days). Melatonin significantly (p less than 0.05) lengthened the duration of preimplantation in both intact (277 +/- 59 days) and SCNx (265 +/- 64.7 days) skunks when compared to all other groups. These data are not consistent with the hypothesis that the SCN are required for melatonin to exert its influence on timing of implantation in the spotted skunk.

Animals↗

Use of histamine2-receptor blocking agents and sucralfate in a health maintenance organization following continued clinical pharmacist intervention.

The effect of continued clinical pharmacist intervention on the proper use of histamine2-receptor blocking agents and sucralfate in a health maintenance organization was studied. New prescriptions written for cimetidine, famotidine, ranitidine, and sucralfate in January 1986 (preintervention), October 1986 (postintervention), and February 1987 (follow-up), and the medical record of each patient were analyzed for appropriateness using approved criteria. Clinical pharmacist intervention occurred throughout the study. There was a significant decrease (p less than 0.005) in the rate of inappropriate prescribing in the postintervention (42.4 percent) and follow-up (48.7 percent) periods versus the preintervention period (81.5 percent), and in the average number of refills authorized per patient in the postintervention (1.2 +/- 1.54) and follow-up (1.3 +/- 1.00) periods versus the preintervention period (3.0 +/- 3.67). There were no significant differences in these areas between the postintervention and follow-up periods (p greater than 0.05). A benefit-to-cost ratio of 4.3:1 was generated from this study which demonstrated that continued clinical pharmacy interventions can have a positive, cost-effective impact on the proper use of these commonly prescribed medications.

Adolescent↗

Changes in endometrial vascular permeability during the periimplantation period in the ferret (Mustela putorius).

A highly localized increase in permeability of uterine blood vessels in the immediate vicinity of implanting blastocysts was first detected on the morning of the 12th day of pregnancy (290 h post coitum). The amount of extravasated dye which accumulated at implantation sites continued to increase through the evening of Day 13 (321 h p.c.). Blastocyst expansion, as indicated by small uterine swellings, preceded a detectable change in vascular permeability by about 10 h, suggesting that the timing of increased permeability is closely associated with initial blastocyst attachment to the uterine epithelium. The results do not support the hypothesis that prostaglandins are required for increased uterine vascular permeability as two doses of indomethacin (4 and 8 mg/kg body wt) administered 5 times/day failed to decrease endometrial vascular permeability. However, the 8 mg dose did cause a significant reduction in size and number of uterine swellings and delayed or inhibited attachment of the trophoblast to the uterine epithelium in 2 of 5 ferrets. These findings suggest that prostaglandins play an important role in the process of implantation that is unrelated to decidual formation as the ferret is an adeciduate species.

Animals↗

Morphological changes in the blastocyst of the western spotted skunk during activation from delayed implantation.

Blastocysts collected from the spotted skunk during delay of implantation, early activation and late activation demonstrate three-tiered growth and developmental changes. The slow-growing blastocyst from the several months of delay is small (less than 1.1 mm) with a rounded inner cell mass consisting of clusters of rounded, lipid-filled cells. During the several days of early activation, the lipid in both inner cell mass and trophoblast diminishes, polyribosomes increase in number, and the endodermal layer differentiates as the blastocyst grows (1.2-1.6 mm). At activation the inner cell mass flattens, becomes uncovered by polar trophoblast, and forms a disc of columnar epiblast cells. The blastocyst expands rapidly during the last 24-48 h prior to implantation to 1.7-2.0 mm, and the trophoblast becomes cuboidal with a marked endocytotic apparatus. The morphological evidence, together with previous studies of protein and RNA synthesis, suggests a tooling-up period during early activation with progressive increases in rates of growth and differentiation in the last hours as implantation approaches.

Animals↗

Evidence for pineal involvement in timing implantation in the western spotted skunk.

Experiments were designed to test the hypothesis that the pineal gland mediates the effects of changing day length and thereby synchronizes implantation within the population of spotted skunks. Intact, sham-superior cervical ganglionectomized (SCGx), SCGx, and SCGx/bilateral orbitally enucleated (B1) pregnant skunks were subjected to a natural photoperiod, and the duration of the preimplantation period was monitored. In a second set of experiments, melatonin was administered to pregnant skunks via two methods to determine whether or not this pineal hormone would mimic the effects of short day photoperiods or B1 on duration of the preimplantation period. Bilateral SCGx, which presumably denervated the pineal, had no effect on the average duration of the preimplantation period (232 +/- 57 vs. 199 +/- 6 days). However, SCGx reversed the inhibitory effect of B1 on implantation as SCGx/B1 animals had an average duration of the preimplantation period that was not significantly different from intact controls (262 +/- 46 vs. 214 +/- 11 days) but was significantly shortened in comparison to enucleated animals (262 +/- 46 vs. 316 +/- 45 days). Melatonin significantly lengthened the duration of the preimplantation period in animals receiving either daily afternoon injections of melatonin or those receiving melatonin Silastic capsule implants. These results are consistent with the hypothesis that the pineal gland synchronizes photoperiod-induced blastocyst implantation in the western spotted skunk.

Animals↗

Role of the corpus luteum in controlling implantation in mustelid carnivores.

The studies that have been reviewed in this paper indicate that the ovaries of mustelids play an essential role in induction of blastocyst implantation. Moreover, the data suggest that CL are the primary source of a hormone, which acts in conjunction with progesterone, to initiate the chain of events leading to implantation. The identity of this luteal factor is unknown; however, existing evidence suggests that it may not be a steroid.

Animals↗

Accumulation of RNA in blastocysts during embryonic diapause and the periimplantation period in the western spotted skunk.

The in vivo incorporation of 3H-uridine into RNA was studied in delayed implanting and activated blastocysts obtained from 33 western spotted skunks. 3H-uridine was incorporated into RNA by all blastocysts; however, significantly more label was incorporated as blastocyst diameter increased. Activated blastocysts with diameters of 1.6 mm or greater on average incorporated 65 times more 3H-precursor in 5 hr than diapausing blastocysts with diameters of 1.1 mm or less. Polyadenylated RNA was likewise synthesized by delayed implanting and activated skunk blastocysts; however, the proportion of polyadenylated RNA synthesized by the former was greater than in the latter (43.9% vs. 27.5%). Our data suggest that the transition from embryonic diapause to fully activated blastocysts first occurs gradually for several days before entering a 1-2-day period of rapid development characterized by an abrupt increase in RNA accumulation.

Animals↗

Presence of an inhibitor of plasminogen activator in uterine fluid of the western spotted skunk during delayed implantation.

This study was designed to test the hypothesis that protease inhibitors would be present in uterine fluids of the western spotted skunk during the period of delayed implantation but would decline or disappear during implantation. Blood plasma, uterine flushings and medium from endometrial cultures were collected 40-70 days preimplantation (Stage 1), 20-25 days preimplantation (Stage 2), 1-3 days preimplantation (Stage 3) and 1-5 days postimplantation (Stage 4). Protein content of the flushings was low in Stage 1 (12.9 +/- 3.0 micrograms), increased slightly (21.8 +/- 6.4 micrograms) during Stage 2 and increased markedly in Stages 3 (99 +/- 44.8 micrograms) and 4 (256 +/- 150 micrograms). No protease activity was detected in any fluid at any stage. Uterine flushings, however, contained an inhibitor of plasminogen activator (PA) of apparent molecular weight approximately 70,000. The total amount of inhibitor increased as pregnancy progressed, but specific inhibitory activity was highest during Stages 1 and 2 (1.7 and 3.2 units/micrograms protein, respectively) and declined in Stages 3 and 4 (1.2 and 0.6 units/micrograms protein, respectively). The inhibitor was present in medium from uterine cultures but was low in plasma. All of the fluids could inhibit trypsin, but not plasmin. The antitrypsin activity in uterine fluids is attributed to plasma transudate. The results demonstrate an inhibitor of PA in skunk uterine fluids but show no temporal relationship between its presence and the period of obligate delay of implantation.

Animals↗