Tamoxifen is an estrogen antagonist on gonadotropin secretion and responsiveness of the hypothalamic-pituitary- adrenal axis in female monkeys.

The selective estrogen receptor modulator, tamoxifen, effectively slows the progression of estrogen-positive breast cancer and reduces the possibility of this cancer developing in women at high risk. Despite the widespread acceptance of tamoxifen as a therapeutic agent for this disease, its effects on other estrogen-dependent pathways, particularly on neural circuits regulating brain function and peripheral hormone secretion, are poorly understood. The present study, using previously ovariectomized rhesus monkeys, examined the effects of tamoxifen, in both the presence and absence of estradiol replacement, on the reproductive and hypo-thalamic-pituitary-adrenal (HPA) axes. In Experiment 1, monkeys randomly assigned to three groups (n = 8 each) were treated with placebo and either two doses of estradiol, two doses of tamoxifen alone, or two doses of tamoxifen plus high-dose estradiol to assess the effects on negative feedback suppression of luteinizing hormone (LH). Both doses of tamoxifen effectively antagonized the negative feedback efficacy of estradiol on LH secretion. In contrast, neither the low- or high-dose tamoxifen alone had any effect on LH secretion, as concentrations during tamoxifen treatments were indistinguishable from those during placebo. In Experiment 2, females were randomly assigned to one of four treatment groups (placebo, n = 6; estradiol, n = 5; tamoxifen only, n = 5; or tamoxifen plus estradiol, n = 6) to assess the effects on glucocorticoid negative feedback and pituitary and adrenal responsiveness to exogenous corticotropin- releasing hormone (CRH). Tamoxifen also antagonized the facilitating effects of estradiol on basal and CRH-induced ACTH and cortisol secretion. However, this antagonism produced basal and CRH-stimulated cortisol and ACTH concentrations that were lower than placebo-treated females. Interestingly, tamoxifen in the absence of estradiol produced a similar diminution in ACTH and cortisol response. These data suggest that, in the presence of estradiol, tamoxifen not only antagonized estrogenic facilitation of HPA responsivity but also actually attenuated the response compared with the placebo-treatment condition. Taken together, these data indicate that tamoxifen acts as an estrogen antagonist on the neural circuits controlling the neuroendocrine regulation of the hypothalamic-pituitary-ovarian and adrenal axes in ovariectomized macaque females.

A comparison of the effects of local analgesic solution in the navicular bursa of horses with lameness caused by solar toe or solar heel pain.

We hypothesised that analgesia of the navicular bursa is not selective for the navicular apparatus; and that solar pain in some horses can be temporarily abolished or attenuated by analgesia of the navicular bursa. To test this hypothesis, we caused lameness in horses by inducing pain in the dorsal margin or the angles of the sole and then evaluated the ability of a local analgesic solution administered into the navicular bursa to attenuate lameness. The response of horses with solar pain in the dorsal or palmar aspect of the foot to 3.5 ml local analgesic solution administered into the navicular bursa was examined. Lameness was induced in 6 horses by creating solar pain in the dorsal aspect of one forefoot and, at another time, the palmar aspect of the other forefoot, with set-screws inserted into a custom-made shoe. Horses were videotaped trotting before and after application of set-screws and after administering 3.5 ml local analgesic solution into the navicular bursa. Lameness scores were assigned by examining videotaped gaits. Scores were significantly lower (P<0.05) for all horses with set-screws applied to the dorsal margin of the sole after administration of local analgesic solution into the navicular bursa. In conclusion, analgesia of the navicular bursa was less effective in desensitising the angles of the sole than in desensitising the dorsal margin of the sole. Pain arising from the sole should not be excluded as a cause of lameness when lameness is attenuated by analgesia of the navicular bursa.

A comparison of the effects of two volumes of local analgesic solution in the distal interphalangeal joint of horses with lameness caused by solar toe or solar heel pain.

The response of horses, with solar pain in the dorsal or palmar aspect of the foot, to 6 or 10 ml local analgesic solution administered into the distal interphalangeal (DIP) joint was examined. Lameness was induced in 7 horses by creating solar pain in the dorsal aspect of one forefoot and, at another time, the palmar aspect of the other forefoot with set-screws inserted into a custom-made shoe. Horses were videotaped trotting before and after application of set-screws and, in separate trials, after 6 or 10 ml local analgesic solution was administered into the DIP joint. Lameness scores were assigned by examining videotaped gaits. Scores were significantly lower (P < 0.05) for horses with set-screws applied to the angles of the sole and receiving 10 ml, but not 6 ml, local analgesic solution into the DIP joint. Scores were significantly lower (P < 0.05) for all horses with set-screws in the dorsal margin of the sole receiving either volume of local analgesic solution. Analgesia of the DIP joint was less effective in desensitising the angles of the sole than in desensitising the dorsal margin of the sole, and 10 ml local analgesic solution was more effective than 6 ml in desensitising these regions. The response of horses with solar pain to local analgesic solution in the DIP joint was influenced by the volume administered and the region of sole affected.

Consequences of the presence of the mother or unfamiliar adult female on cortisol, ACTH, testosterone and behavioral responses of periadolescent guinea pigs during exposure to novelty.

Periadolescent guinea pigs were exposed to a novel environment for 10, 30, 60, or 90 min while either alone or with either their biological mother or an unfamiliar adult female. Both classes of females reduced plasma cortisol, ACTH and vocalization responses to the novel environment, and did so to an equivalent degree. However, behavioral interactions with the mother and unfamiliar female differed considerably. The offspring exhibited more defensive behavior and were the targets of more agonistic acts when with the unfamiliar female, though one agonistic behavior, kicking, was observed more often in tests with the mother. Males displayed more social/courtship behavior and tumescence with the unfamiliar female. Further, males exhibited higher plasma testosterone levels when with the unfamiliar female than when either with the mother or alone. These data in conjunction with earlier findings suggest that the ability of unfamiliar adult females to moderate hypothalamic-pituitary-adrenal activity fully emerges during the periadolescent period, is not dependent on nurturant behavior by the adult female, and may facilitate the redirection of social behavior from the mother to unrelated adults. Further, mothers appear to inhibit maternally directed sexual behavior and plasma testosterone elevations in their periadolescent sons, effects which likely serve to inhibit inbreeding.

Functional expression of the pore forming subunit of the ATP-sensitive potassium channel in Saccharomyces cerevisiae.

We have expressed the pore-forming subunits (Kir 6.1 and Kir 6.2) of the mammalian ATP-sensitive potassium channel in a potassium-transport deficient yeast strain (trk1 trk2). Functional expression of Kir 6.2 and Kir 6.1 can complement growth deficiency weakly and strongly respectively of the yeast strain on low-potassium medium. Mutations of Kir 6.2 that abolish ATP sensitivity (K185Q, I182Q) and enhance trafficking to the plasma membrane surface (Kir 6.2DeltaC36) lead to significantly better growth rescue. Growth rescue of Kir 6.1, Kir 6.2 and the above mutants can be inhibited by pharmacological agents (cesium ions, phentolamine and quinine) known to decrease channel activity by direct interaction with the pore forming subunit. Thus we have developed a system in yeast that can report both loss and gain of function mutations in these subunits and pharmacological interventions.

Effects of analgesia of the distal interphalangeal joint or palmar digital nerves on lameness caused by solar pain in horses.

OBJECTIVE: To determine if pain of the dorsal margin of the sole in horses can be attenuated by anesthesia of either the distal interphalangeal (DIP) joint or the palmar digital (PD) nerves. STUDY DESIGN: A unilateral forelimb lameness was induced by creating solar pain. Response to administration of local anesthetic or saline solution into the DIP joint and to administration of local anesthetic around the PD nerves was evaluated. ANIMALS: Six horses. METHODS: Lameness was induced by creating pressure on the dorsal margin of the sole by screwing set-screws into a nut welded to the inside of each branch of a shoe. Gaits were evaluated before and after application of set-screws and after a local anesthetic or saline solution was administered into the DIP joint and, in a second trial, after a local anesthetic was injected around the PD nerves. Gaits recorded on videotape were evaluated, and lameness scores were assigned to each gait. RESULTS: Lameness scores were high after application of set-screws and remained high after saline solution was administered into the DIP joint. Scores decreased significantly (P < or = .05) after a local anesthetic was administered into the DIP joint or around the PD nerves. CONCLUSIONS: Analgesia of the DIP joint or the PD nerves desensitizes at least a portion of the sole. CLINICAL RELEVANCE: Pain arising from the sole should not be excluded as a cause of lameness when lameness is attenuated by analgesia of the DIP joint or PD nerves.

Comparison of the effects of the mother and an unfamiliar adult female on cortisol and behavioral responses of pre- and postweaning guinea pigs.

In the guinea pig, the presence of the mother, but not littermates, has been found to inhibit hypothalamic-pituitary-adrenal (HPA) responses during brief (30-60 min) exposure to novel surroundings both prior to and several weeks following the completion of weaning. In the present study, we found that an unfamiliar adult female inhibited plasma cortisol and vocalization responses of pre- and postweaning guinea pigs during a 60-min exposure to a novel environment. However, the presence of the mother still had a greater effect on the cortisol levels of the young, at least during the preweaning period. The moderating influence of the unfamiliar adult female on vocalizations and cortisol levels occurred despite behavioral interactions, such as heightened aggression and sexual behavior, not seen during tests with the mother. It is suggested that the unfamiliar adult female's effectiveness in reducing HPA activity during exposure to novelty may facilitate the change in patterns of social interaction occurring in recently weaned animals.

The C-terminus of eRF1 defines a functionally important domain for translation termination in Saccharomyces cerevisiae.

Translation termination in eukaryotes is mediated by two release factors, eRF1 and eRF3, which interact to form a heterodimer that mediates termination at all three stop codons. By C-terminal deletion analysis of eRF1 from the yeast Saccharomyces cerevisiae, we show that the extreme C-terminus of this 437-amino-acid protein defines a functionally important domain for translation termination. A strain encoding eRF1 lacking the C-terminal 32 amino acids is not viable, whereas deletion of the C-terminal 19 amino acids is viable but shows a termination defect in vivo causing an enhancement of nonsense suppression. Using a combination of two-hybrid analysis and in vitro binding studies, we demonstrate that deletions encompassing the C-terminus of eRF1 cause a significant reduction in eRF3 binding to eRF1. All of the C-terminally truncated eRF1 still bind the ribosome, suggesting that the C-terminus does not constitute a ribosome-binding domain and eRF1 does not need to form a stable complex with eRF3 in order to bind the ribosome. These data, together with previously published data, suggest that the region between amino acids 411 and 418 of yeast eRF1 defines an essential functional domain that is part of the major site of interaction with eRF3. However, a stable eRF1:eRF3 complex does not have to be formed to maintain viability or efficient translation termination. Alignment of the seven known eukaryotic eRF1 sequences indicates that a highly conserved motif, GFGGIGG/A is present within the region of the C-terminus, although our deletion studies suggest that it is sequences C-terminal to this region that are functionally important.

Ketoprofen concentrations in plasma and milk after intravenous administration in dairy cattle.

OBJECTIVE: To determine plasma and milk concentration-time profiles and pharmacokinetic variables after i.v. administration of ketoprofen to lactating dairy cows. DESIGN: Cows received a single i.v. bolus of ketoprofen (3.31 mg/kg of body weight). Blood and milk were collected at 0, 5, 10, 15, 25, 40, 60, 90, 120, 180, 240, 360, and 480 minutes. Ketoprofen concentrations in plasma and milk were determined. ANIMALS: 6-clinically normal lactating Holstein cows. PROCEDURE: Plasma and milk samples were processed by solvent extraction, and ketoprofen concentrations were determined, using high-performance liquid chromatography with octadecyl silane reverse-phase guard and analytic columns. A computer polyexponential curve-stripping program was used to fit ketoprofen concentration-time data and to calculate pharmacokinetic variables. RESULTS: The lower limit of detection for ketoprofen in plasma was 18 ng/ml; the lower limit of quantification was 60 ng/ml. The lower limit of detection for ketoprofen in milk was 27 ng/ml; the lower limit of quantification was 90 ng/ml. Plasma ketoprofen concentration-time curves best fit an open two-compartment pharmacokinetic model. Harmonic mean apparent volume of distribution at steady state was 0.11 (range, 0.095 to 0.13) L/kg, elimination half-life was 0.49 (range, 0.40 to 0.67) hour, and total clearance was 0.17 (range, 0.14 to 0.19) L/kg/h. Ketoprofen was detected in some milk samples, 10 to 120 minutes after administration, but all concentrations were below the limit of quantification. Adverse effects were not observed in cows given ketoprofen. CONCLUSIONS: The elimination of half-life for ketoprofen is short, and low concentrations of ketoprofen can be detected in normal milk, after i.v. treatment of cattle with ketoprofen. Milk and meat from cattle treated i.v. with ketoprofen should not be an important drug residue risk if appropriate withholding periods are used.

Cellulases and hemicellulases of the anaerobic fungus Piromyces constitute a multiprotein cellulose-binding complex and are encoded by multigene families.

More than 80% of the extracellular Avicelase, endoglucanase, xylanase and mannanase activities of the anaerobic fungus Piromyces were associated with a cellulose-binding complex. The complex was composed of at least 10 polypeptides ranging in size from 190 kDa to 50 kDa, and contained numerous endoglucanases, xylanases and mannanases. Multiple genes encoding each of these activities were isolated from an expressing cDNA library.

An experimental study of the anatomy of the tubules of the human kidney and its relation to calculus formation.

During a period of 28 years 6 clinical observations concerning humans and a dog indicated that micro-calculi can form and acquire laminated growth at the base of the papillae in the renal medulla. In the subsequent research, plastic casts were made of the distal tubules of human post mortem kidneys. Study of these showed that acute angle and Z-bends were more prevalent at the bases of the papillae in the lower calices. Macro-photographs revealed that kinks in the wall of the tubule can reduce the lumen at these points. The combined evidence suggests that crystals and cellular debris may become impacted at these sites and form the nucleus of a stone.

An osteotomy technique in the treatment of staghorn calculus.

A technique was devised for dividing the staghorn calculus with an osteotome after obtaining access to the stone by means of a partial nephrectomy aided by renal hypothermia. The advantages of the procedure are a better control of haemorrhage and a clean cut in the stone with removal of predetermined fragments and a diminished splinter formation. In 17 operations there were no complications of haemorrhage nor urinary fistula and, as yet, no recurrence of stone.