A new prostaglandin E1 analogue (CL115, 574): I. Antisecretory and cytoprotectige effects in the rat

The effect of a new analogue of PGE₁ (CL115, 574) on gastric acid secretion, mucus secretion and protection against stress and indomethacin- induced ulcers were studied in the rat. CL115, 574 was more potent than PGE₁ and cimetidine in inhibiting acid secretion. CL115, 574 protected against the development of stress and indomethacin-induced ulcers prevented the indomethacin-induced decrease in hematocrit at an ED₅₀ (3 μg/kg) far below the antisecretory ED₅₀ (1 mg/kg). While the inhibiting acid secretion, CL115, 574 increased the volume of gastric secretion indicating a stimulation of nonparallel cell secretion by the rat stomach. In addition the compound stimulated the secretion of mucus into the gastric juice. On the basis of its potency as an inhibitor of acid secretion and these additional effects which are indicative of cytoprotective activity, CL115, 574 should be further studied as a possible anti-ulcer agent in man.     

Naloxone hyperalgesia and stress-induced analgesia in rats

Since past studies concerning the effects of naloxone on nociception have yielded inconclusive findings, the variables of pain test, baseline sensitivity, and stress condition were examined. Within a pure-bred strain of rats, consistent individual differences did not occur. All three measures of pain responsiveness demonstrated hyperalgesic effects of naloxone, but they differed in their capacity to reflect the effects of analgesia produced by continuous or intermittent electrical shock. By some measures, naloxone reversed the stress-induced analgesia due to intermittent shock; it did not influence the analgesia produced by continuous stress. The data support a model of pain inhibition involving both opioid and non-opioid systems and suggest that the hyperalgesic effects of naloxone can sometime gives rise to erroneous conclusions concerning apparent naloxone-reversability of putative analgesic procedures.     

Inhibition of an IUD-induced precocious luteolysis by prostaglandin E1 (PGE1) in sheep

Fifteen ewes were assigned as they came into estrus to one of three randomized treatment groups: 1. Sham IUD + Vehicle, 2. IUD + Vehicle or 3. IUD + PGE₁ in vehicle. An IUD was inserted adjacent to the luteal-bearing ovary on day 3 postestrus. Prostaglandin E₁ (500 μg) in vehicle (Na₂CO₃) or vehicle was given intrauterine through an indwelling uterine cannula every four hours from day 3 postestrus until ewes returned to estrus. Precocious estrus was induced in both the sham IUD groups receiving vehicle. Prostaglandin E₁ prevented an IUD-induced premature luteolysis based on daily concentrations of progesterone in peripheral blood and the interestrous interval. It is concluded that an IUD-induced premature luteolysis is not necessarily via physical distention by the IUD. It is also concluded that chronic intrauterine infusions of PGE₁ can prevent an IUD-induced premature luteolysis.     

Effects of prostaglandin E2 (PGE2) on estradiol-17β-induced luteolysis in the nonpregnant ewe

Fifteen ewes were assigned as they came into estrus to the following randomized treatment groups: 1) Vehicle (1 ml corn oil + vehicle Na₂CO₃ buffer), 2) Estradiol-17β + vehicle and 3) Estradiol-17β + PGE₂ (500 μg) in Na₂CO₃ buffer (5 ewes/treatment group). Prostaglandin E₂ was given through an intrauterine cannula every four hours from days 8 through 15 postestrus. PGE₂ prevented a luteolytic dose of estradiol-17β given on days 9 and 10 from causing a precious luteolysis. PGE₂ maintained concentrations of progesterone in peripheral blood (days 8 through 15) and weights and concentrations of progesterone in corpora lutea on day 15 postestrus of ewes receiving estradiol-17β. It is concluded that chronic intrauterine infusions of PGE₂ can prevent an estradiol-17β-induced premature luteolysis.     

Auto and heterotransplantation of the ovary to the uterus or parotid region in beef cows and prepuberal heifers

Eighteen mature estrous cycling beef cows and 9 prepuberal heifers were stratified by breed, age and weight to determine the effect of ovary-transplantation to a proximal site (right uterine horn) (U) and distal site (parotid region) (P) upon ovarian activity. Active ovaries (AO), ovaries with the corpus luteum (CL), were autotransplanted to the myometrium of the U in 3 cows and to the muscles of the P in 2 cows and their inactive ovaries (IO), ovaries without a CL, remained. Active ovaries of 6 cows were removed and heterotransplanted to 6 prepuberal heifers and their 10 were heterotransplanted to the U or the parotid (3). Six heifers received either a mature AO in the U or in the parotid. Three heifers were ovariectomized and their ovaries were heterotransplanted to 6 cows, 3 per site. Cows and heifers were slaughtered randomly 2 months after surgery and their ovaries were collected for microscopic and histological analysis. The transplants were successfully accomplished in 94% of the cows and in 83% of the heifers. Both of the unsuccessful heterotransplantations were located in the uterus. More estrous activity was found (P<.025) in cows than in heifers with their own ovaries in situ . All prepuberal ovaries in situ showed follicular development when mature AO were transplanted to either the U or parotid. The same trend was found in prepuberal ovaries transplanted to mature cycling cows. Cows with an IO in situ and AO transplanted to either site had more estrous activity than did ovariectomized cows with an IO transplanted to either site. Pregnancy rates in mature cycling cows with an least one ovary in situ were higher (P<.005) in cows with an ovary in the parotid region than cows with an ovary transplanted to the uterus.     

Pheromone-stimulated locomotory and orientation responses in the American cockroach

Males of Periplaneta americana respond to the sex pheromone secreted by females with increased locomotion and positive chemo-orientation. Both sexes orient toward aggregation pheromone without an increase in locomotion. Immediately following removal of one antenna males exhibit ‘circus’ movements, but after 2 days orient toward pheromones; a bi-modal mechanism of chemo-orientation is proposed. Cockroaches turn away from air currents, but orient toward air currents carrying sex or aggregation pheromone, suggesting the possibility of up-wind orientation. Antennae deflect upward and outward when pheromone is first perceived; the head then moves toward the pheromone source. Following removal of one antenna, the pattern of head and antennal movement changes in a manner which enhances the sweeping of the intact antenna.     

Cyclooxygenase and lipoxygenase pathways in the preimplantation rabbit uterus and blastocyst

We have measured by radioimmunoassay the concentration and production of 5(S)-hydroxy-6,8,11,14-eicosatetraenoic acid (5-HETE), a metabolite in the lipoxygenase pathway, and PGs in different uterine compartments, and blastocysts during the preimplantation period in the rabbit. The production is defined as the synthesis minus the metabolism for a defined period of time. The pattern of uterine PGF production on days 5–6.5 was quite similar for the whole uterus and the myometrium showing a peak production on Day 6. The concentration and production of PGF were always higher in the endometrium. While significant production of PGE was noticed in the whole uterus on days 5–6 and in the myometrium on Day 6, the endometrium showed some production on these days. On the contrary, absolutely no production of this PG was observed in the endometrium on Day 6.5. The concentration and production of 6-keto-PGF_1α were always lower in the endometrium than those observed in the myometrium or the whole uterus. While highest production of this PG was found to be on Day 6.5 in the whole uterus and on Day 5 in the endometrium, the production in the myometrium remained constant on all days examined. The production of 5-HETE in the endometrium was noticeable on Days 5–6.5, in the whole uterus on Days 5 and 6.5, and in the myometrium only on Day 6.5. However, the concentrations of 5-HETE showed a tendency to be higher at 2 h than at 0 h in these compartments on Days 5–6.5. Furthermore, a linear increase in 5-HETE levels both 0 h and 2 h was observed in the endometrium on Days 5–6.5; no such differences in mean 5-HETE level was noted in the whole uterus or myometrium on any of these days. The production of 5-HETE in the blastocyst was noted only on Day 5. The results not only demonstrate the presence of both the cyclo-oxygenase and the lipoxygenase pathways in the preimplantation rabbit uterus and blastocyst, their differential operation in various compartments of the uterus on various days of early pregnancy suggests an integrated role for these mediators in embryo-uterine interaction during implantation.     

Central cardiovascular and thermal effects of prostaglandin F2 alpha in rats.

Administration of PGF2 ALPHA (0.2--6.4 micrograms) into the lateral cerebral ventricle (i.c.v.) induced dose-dependent increases in blood pressure, heart rate and body temperature in urethane-anaesthetised rats, but had no effect on these parameters when the same dose range was administered intravenously. Peripheral pretreatment with sodium meclofenamate (50 mg/kg s.c.) shifted all the dose-response curves for PGF2 alpha (i.c.v.) to the left, but indomethacin (50 mg/kg s.c.) did not significantly affect those changes. Central pretreatment with sodium meclofenamate or indomethacin (1.25 mg per rat i.c.v.) failed to modify significantly the effects of centrally administered PGF2 alpha. The results support previous suggestions that PGF2 alpha may participate in the central control of the cardiovascular and thermoregulatory systems, and also suggest that there may be differences in the sites and/or modes of action between sodium meclofenamate and indomethacin.     

Treatment of chronic kidney transplant rejection with prostacyclin — reduction of platelet deposition in the transplant; Prolongation of platelet survival and improvement of transplant function

8 patients with chronic kidney transplant rejection were treated intravenously with prostacyclin during 5 days. This treatment seemed to have a beneficial effects as measured by platelet deposition in the transplant and prolongation of platelet survival. In the majority of cases the transplant function improved. A longer duration of this therapeutic effect could be a new way for the long-term treatment of chronic kidney transplant rejection.     

A study of the effect of caffeine upon excision repair of damaged DNA.

Cultured mammalian cells incur damage to their DNA when exposed to ultraviolet light or adduct-producing mutagens such as 4-nitroquinoline-1-oxide (4NQO). At least two processes are important in repair of such damage: post-replication repair and excision repair. Many researchers have reported that caffeine inhibits the former process, which occurs in connection with semiconservative DNA replication, especially in rodent cell lines such as mouse lymphoma or Chinese hamster. Excision repair is not generally considered caffeine-sensitive, although the data are somewhat conflicting because some studies had used rodent cells, which show little or no excision repair, or human cells in which alternate repair processes may have been operating.Human peripherhal blood lymphocytes from healthy donors were treated with UV light or 4NQO in order to produce pyrimidine dimers or adducts. Caffeine at concentrations of 0.75–3.0 mM was included in some cultures. The cells treated with caffeine were incubated for 90 min prior to mutagen treatment and for the entire period thereafter until cell harvests. [³H]Thymidine was added and the uptake quantitated as a measure of DNA repair. DNA replication was inhibited by hydroxyurea, so that only excision repair was measured by this method. Separate plates of cells not exposed to mutagens exhibited negligible or low thymidine uptakes.Following harvest, the cells were lysed and the DNA extracted. The DNA released was measured spectrophotometrically and then placed into liquid-scintillation counter (LSC) vials for measurement of incorporated radioactivity. Resulting cpm/μ DNA were compared for cells with and without caffeine. Lymphocytes from patients with systemic lupus erythematosus (SLE), who previously had demonstrated reduced levels of excision repair under these conditions, were also tested with caffeine. Caffeine did not inhibit repair by normal lymphocytes and the reduced repair seen in the SLE patients was not further reduced in its presence.In a series of pulse-chase experiments, some cells were treated with 4NQO and allowed to incubate with [³H]thymidine for 3 h and were harvested at the end of this period, while others were given a 13-h chase i n cold thymidine before harvest. The cpm/μg DNA for both groups were virtually identical, both in the presence and absence of 2.0 mM caffeine.     

Central cardiovascular effects of baclofen in spontaneously hypertensive rats

This study was conducted to investigate the effects of centrally administered baclofen on blood pressure and heart rate in conscious spontaneously hypertensive (SHR) and normotensive Wistar-Kyoto (WKY) rats. Administration of baclofen (1.0 microgram/kg) into the lateral cerebral ventricle (icv) produced an increase in mean arterial pressure (MAP) in both SHR and WKY rats. The increase in MAP was significantly lower in SHR (13 +/- 3 mmHg) when compared with WKY (27 +/- 5 mmHg). The changes in heart rate (HR) were variable, from no change to a very small increase and did not differ significantly between SHR and WKY rats. The ability of baclofen to interfere with baroreceptor reflexes was also tested in separate experiments. In SHR, icv injection of baclofen (1.0 microgram/kg) significantly suppressed the pressor response and bradycardia evoked by phenylephrine 3.0 micrograms/kg iv, whereas in WKY, the pressor and HR responses to similar injections of phenylephrine were not affected by icv baclofen. Similarly, baclofen treatment modified hypotensive response and reflex tachycardia induced by nitroprusside (10.0 micrograms/kg) iv in SHR but not in WKY rats. Administration of sympathetic ganglionic blocker hexamethonium (HEX; 25 mg/kg) iv produced an equivalent decrease in MAP between SHR and WKY following icv injection of baclofen (1.0 microgram/kg). These results suggest that the effects of baclofen on the baroreceptor reflexes in SHR may not be mediated by a change in peripheral sympathetic tone.     

ACTH1-24 effects on d-amphetamine self-administration and the dynamics of brain dopamine in rats

Experiments aimed at determining the neural basis of reward have previously focused on the role of neurotransmitters and have only recently begun to investigate the role of peptides. The present experiment investigated the effect of ACTH1-24 on d-amphetamine self-administration in rats. Animals were trained daily (8 hour sessions) to press a lever which activated a system that administered 0.125 mg/kg of intravenous amphetamine. After achievement of a stable self-injection frequency, subjects were injected SC with 10, 20 or 40 micrograms/80 microliters ACTH1-24 immediately prior to placement in the apparatus. The 20 micrograms and 40 micrograms doses of the peptide fragment induced a statistically significant attenuation of d-amphetamine self-injection which lasted for 2 days. Control rates of responding were achieved by 5 to 10 days after the peptide treatment. An experiment was conducted to evaluate possible neuromodulatory effects of the peptide fragment. Twenty-four hr after ACTH1-24, HVA was elevated in the caudate. When both apomorphine and ACTH1-24 were administered, the combination lowered HVA in the caudate to a greater degree than apomorphine alone. The peptide fragment, when combined with haloperidol, attenuated the haloperidol-induced increases of DOPAC and HVA in both the caudate and nucleus accumbens. It was tentatively concluded that the neuromodulatory action of ACTH1-24 on dopaminergic neurons may result in an increase in the rewarding quality of d-amphetamine, thus rendering control level self-infusions superfluous.(ABSTRACT TRUNCATED AT 250 WORDS)     

Correlation of the plasma tyrosine to phenylalanine ratio with energy intake in self-selecting weanling rats

The relationship between changes in blood plasma amino acids and the quantity of protein and energy self-selected by the weanling rat, simultaneously offered two diets varying only in gluten (15 and 55%) concentration, was examined. Gluten and energy intakes were manipulated by additions of lysine, arginine or ammonia to gluten. In two experiments groups of ten weanling rats were fed the diets for a two week or four week period and food intake selection recorded. Blood samples were obtained between 0900–1100 hr at the end of the two week or four week period. Correlation coefficients of protein intake with the plasma TRP/NAA (Tyr+Phe+Leu+Val+Ile) ratios were ?0.97 and ?0.98, and of energy intake with TYR/PHE ratios were 0.77 and 0.70 in experiments 1 and 2, respectively. It is suggested that the plasma TRP/NAA and TYR/PHE ratios reflect the mechanisms regulating protein and energy intakes, respectively.     

Effects of prostacyclin on ultrastructural features of human platelets in vitro

Examining platelets plasma-membrane by freeze-etching (FE)-technique irregularly placed among the “particles”, some larger formations, called by us “protuberances” can be seen. These protuberances may represent the stomata of the open canalicular system (OCS) and the morphological aspect of an exocytotic process. Addition of prostacyclin to platelet rich plasma (PRP) leads to a decrease of the average value of protuberances which is statistically significant in comparison to control platelets.     

Localization of glycoprotein glycosyltransferases in the Golgi apparatus of rat and mouse testis

Golgi fractions prepared from rat testis have been shown to be enriched in the following glycoprotein glycosyltransferases: N-acetylglucosaminyltransferase, 47-fold, galactosyltransferase, 33-fold, and N-acetylglucosaminide fucosyltransferase, 15-fold. Appreciably lower transferase levels were obtained in other subcellular fractions. In the mouse, Golgi fractions were prepared from testis homogenates, testis cell suspensions and partially purified testis germinal cells; these fractions were also enriched in the above glycoprotein glycosyltransferases. Electron microscopic analysis indicated that a major portion of the total transferase activity was located in the Golgi apparatus of both rat and mouse testis although these experiments could not rule out the possible presence of some transferase activity in other organelles.     

Radiation induced alterations in rabbit aortic prostacyclin formation

Earlier studies have demonstrated very severe vascular lesions occuring after irradiation in human and experimental animals as well. We examined the rabbit aortic PGI₂-formation using the platelet bioassay technique in one year aged rabbits after different irradiation doses and a time course. The findings demonstrate a signficant increase in prostacyclin formation, which might be due to the damage of endothelial cells. This stage is followed by a long lasting severe depression. This behaviour in addition demonstrates a dose dependent manner. The findings of a long lasting decrease in vascular PGI₂-formation might contribute to the understanding of the high incidence of vascular lesions being found at the site of irradiation.     

Morphine is most effective on gastrointestinal propulsion in rats by intraperitoneal route: Evidence for local action

A very low dose of morphine (37.5 μg/kg) entirely delivered into the peritoneal cavity of rats, consistently reduced the transit of a forced charcoal meal through the small intestine (to about 30% of control), but failed to elicit such action in naloxone-pretreated animals or if administered either intravenously or intracerebroventricularly. The same dose of tritium labeled morphine, injected i.v., resulted in brain and small intestine morphine levels respectively 2.7 times higher and 3.9 times lower than in the corresponding tissues of rats injected i.p.. These findings suggest that activation of opiate specific sites located in the gastrointestinal tract can $\text{per se}$ be primarily responsible for the antipropulsive effects of morphine.     

Central cardiovascular effects of mammalian neurohypophyseal peptides in conscious rats

To confirm and extend the results of previous studies which demonstrated central cardiovascular effects of vasopressin in anesthetized rats, we determined blood pressure and heart rate changes for 30 minutes after intracerebroventricular injections of arginine vasopressin, arginine vasotocin and oxytocin in conscious rats. As compared to sham injections, significantly greater increases in either systolic or diastolic blood pressure were noted over the 30 minutes which followed the injection of 0.15, 1.0 or 10.0 nM of either vasopressin or vasotocin. In animals given vasopressin, plasma levels of the peptide were determined. There was a substantial increase in plasma vasopressin only after the highest dose. Overall blood pressure responses to doses of oxytocin as high as 100 nM were not significantly different than sham injections. Heart rate following both vasopressin and vasotocin was increased at 0.15 nM, was initially decreased then increased at 1.0 nM and was substantially decreased after the 10.0 nM dose. There was a significant increase in heart rate at the 10.0 nM and 100 nM doses of oxytocin. Dose response curves for systolic blood pressure and heart rate 20 minutes after injection were similar for vasopressin and vasotocin. We conclude that arginine vasopressin has significant central pressor and tachycardic effects in conscious rats, and it is related, at least in part, to the tail structure of the peptide, which is shared with arginine vasotocin.