Effect of the cathepsin K inhibitor odanacatib administered once weekly on bone mineral density in Japanese patients with osteoporosis--a double-blind, randomized, dose-finding study.

UNLABELLED: The efficacy and safety of oral placebo or odanacatib 10, 25, or 50 mg once weekly for 52 weeks were evaluated in a double-blind, randomized, multi-center study in Japanese female and male patients with osteoporosis. INTRODUCTION: Odanacatib is a selective and reversible cathepsin K inhibitor that decreases bone resorption and increases bone mineral density (BMD). METHODS: The primary efficacy endpoint was percent change from baseline to week 52 in lumbar spine BMD. Secondary endpoints included percent change in total hip, femoral neck, and trochanter BMD and in bone biomarkers after treatment for 52 weeks. RESULTS: In this study, 286 patients [94% female, mean age (SD) 68.2 (7.1) years] were included in the analysis. The least-squares mean percent changes from baseline to week 52 in the groups receiving placebo, 10, 25 and 50 mg of odanacatib for lumbar spine (L1~L4) BMD were 0.5, 4.1, 5.7, and 5.9% and for total hip BMD were -0.4, 1.3, 1.8, and 2.7%, respectively. The changes in femoral neck and trochanter BMD were similar to those at the total hip. Bone turnover markers were reduced in a dose-dependent manner. However, the effects of odanacatib on bone formation markers were less compared with the effects on bone resorption markers. Tolerability and safety profiles were similar among all treatment groups with no dose-related trends in any adverse events. CONCLUSIONS: Weekly odanacatib treatment for 52 weeks increased BMD at the lumbar spine and at all hip sites in a dose-dependent manner and was well tolerated in Japanese patients with osteoporosis.

Finasteride 1 mg has no inhibitory effect on omeprazole metabolism in extensive and poor metabolizers for CYP2C19 in Japanese.

OBJECTIVE: To examine the inhibitory effect of finasteride 1 mg on the metabolism of omeprazole in genetically determined extensive (EMs) and poor metabolizers (PMs) for CYP2C19 in young healthy Japanese male subjects. METHODS: Twenty-four volunteers participated in this study, among whom 12 were homozygous EMs and 12 were PMs for CYP2C19. A single center, controlled, randomized, open, crossover study with a 5 day washout between the two study periods was performed. Each of the six EMs and PMs received a single oral 20 mg dose of omeprazole on day 1 (treatment I). After a 5 day washout period, these subjects received 1 mg of finasteride once a day for three consecutive days, and a single oral 20 mg dose of omeprazole was co-administered on day 3 (treatment II). The 12 other EMs and PMs received treatments I and II in reverse. Plasma samples were collected for up to a 12 hours postdose of omeprazole, and the pharmacokinetic parameters of omeprazole were determined. RESULTS: The geometric mean ratio (GMR) for the AUC((0-12 hr)) of omeprazole when co-administered with finasteride/omeprazole alone is 1.13 (90%CI, 1.03, 1.25) and 0.96 (0.88, 1.05) in EMs and PMs, respectively. Finasteride did not significantly alter C(max), T(max) and t(1/2) in both genotypes. CONCLUSION: Finasteride 1 mg, widely used for the treatment of androgenetic alopecia in men, did not meaningfully increase omeprazole exposure (20 mg) in both EMs and PMs for CYP2C19. These results indicate that finasteride does not meaningfully inhibit CYP2C19 activity in vivo at the dose of 1 mg.

[Recurrent lung cancer with interstitial pneumonia treated with percutaneous radiofrequency ablation].

We performed computed tomography (CT)-guided percutaneous radiofrequency ablation (RFA) for postoperative recurrent pulmonary metastases developed in a 77-year-old man with interstitial pneumonia. He had received left upper segmentectomy with ND 2a nodal dissection. RFA was safely performed for pulmonary metastases in right S6 and left S6. There was no evidence to suggest any deterioration on interstitial pneumonia, including KL 6 and CT findings. Autopsy revealed residual cancer cells in peripheral lesion in 1 of 2 tumors treated by RFA. Although RFA is palliative, it is a promising treatment for local control of pulmonary malignancy in high-risk patients.

Derivatization LC/MS for the simultaneous determination of fatty alcohol and alcohol ethoxylate surfactants in water and wastewater samples.

A new LC/MS method has been developed for the simultaneous measurement, in water and wastewater samples, of all species contained in commercial samples of linear type of alcohol ethoxylate (AE) surfactants including fatty alcohols. The method requires derivatization of the terminal hydroxyl of each surfactant species with 2-fluoro-N-methylpyridinium p-toluenesulfonate, which imparts a permanent cationic charge, allowing all species including the fatty alcohols and those with only one ethoxylate to be effectively detected by electrospray MS. Detection limits of typically <10 ppt for each individual species were attained in treated wastewater, in which total AE concentrations (combination of up to 114 individual species) are not expected to exceed 10 ppb. The method was validated for clean water as well as sewage influent and effluent samples.

Calcium concentration and fertilization by subzonal insemination of a single spermatozoon in mouse oocytes.

The effect of calcium concentration in culture medium on the fertilization of subzonally microinseminated mouse oocytes was examined. Oocytes were injected with a single spermatozoon so that the sperm head was forced to adhere onto the ooplasmic membrane with a micromanipulation technique. For the inseminations, epididymal spermatozoa preincubated in culture medium and those treated with ionophore A23187 were used. Inseminated oocytes were cultured using media with three different calcium concentrations of 1.71, 3.42, and 5.13 mM; 40.0%, 71.6%, and 47.9% of oocytes microinjected with preincubated sperm were fertilized after incubation with those media, respectively. When the oocytes inseminated with ionophore-treated sperm were incubated in media containing 1.71 and 3.42 mM calcium, their fertilization rates were 58.2% and 87.5%. Thus fertility of subzonally microinseminated oocytes was obviously enhanced when cultured in medium with 3.42 mM of calcium, irrespective of being inseminated with preincubated sperm (P < 0.01) or with ionophore-treated sperm (P < 0.005). Some of the microinseminations with preincubated sperm were performed without sperm adhered to the oolemma. In these cases, the incidence of fertilization was not improved by incubating the inseminated oocytes in medium containing 3.42 mM calcium (32.6%) as compared to those incubated in medium with 1.71 mM calcium (28.3%). These results suggest that the concentration of extracelluar calcium exerts an important effect on the progress of fertilization events subsequent to sperm adherence onto the ooplasmic membrane. Almost 80% of the zygotes fertilized via incubation in medium with 3.42 mM of calcium developed into blastocysts after culturing in vitro.

Subzonal insemination with a single spermatozoon using manipulation assisted sperm adhesion onto the ooplasmic membrane in mouse ova.

A simple and successful method of microinjection of a single spermatozoon under the zona pellucida of a mouse oocyte has been developed. A characteristic of this method is that the tip of the sperm injection needle pierces the zona pellucida without touching the ooplasmic membrane. All the ova (277) used for this series of experiments had normal morphology after the injection procedure. Spermatozoa preincubated in culture medium for capacitation and those treated with ionophore A23187 for induction of acrosome reaction were used. In combination with some of these injections, a manipulation assisting the adhesion of the sperm head onto the ooplasmic membrane was employed. The fertilization rate (67.3%) of the ova injected with the ionophore-treated sperm using the sperm-adhesion treatment was significantly higher (P less than 0.005) than that obtained by the injection of the preincubated sperm without applying the adhesion treatment (23.6%). All three of the recipients that received the 24 fertilized ova became pregnant and gave birth to 11 offspring (45.8%). The inseminations performed with the sperm-adhesion treatment using the immotile sperm from the preincubated population and/or those from the ionophore-treated population did not result in fertilization in any case. These results suggest that the fertilization rate of subzonal insemination with motile ionophore-treated sperm can be improved by applying the sperm-adhesion treatment and that sperm motility might be involved in the establishment of fertilization, even after the adhesion of the sperm head with the mouse ovum membrane.

Effects of aminoglutethimide phosphate administration on the incidence of pregnancy and number of live fetuses in superovulated rats.

This study examined the effects of aminoglutethimide phosphate (AGP) administration on the incidence of pregnancy and number of live fetuses in superovulated rats. Immature (31- to 33-day-old) young (8- to 9-week-old) and (14- to 16-week-old) rats were treated with 20 or 40 IU of PMSG and the same dosses of hCG at 52 to 54 hour-intervals (GTH rats). One half of the GTH rats were injected with 10 mg AGP twice a day for the first 3 days of pregnancy (AGP rats). Pregnancy rates of all AGP rats (95 to 100%) were much higher than the GTH rats (12.7). Young and adult AGP rats treated with 20 IU of GTH had 14.7 and 15.3 live fetuses per dam, respectively. These numbers of live fetuses increased to 17.8 and 18.4 after treatment with 40 IU of GTH. These values were significantly greater than those of the control rats (12.3 to 12.8; P<0.01), but the fetal weights were lower. The results show that AGP administration improves pregnancy rates and the number of live fetuses in young and adult superovulated rats.

Factors affecting the response of the female rat reproductive system to cannabinoids.

Chronic oral administration of either crude marihuana extract (CME) or delta 9-tetrahydrocannabinol (THC) to female Fischer rats for 64-72 days, at a dose approximating heavy usage by humans, reduces food intake by about 8%. Pair-feeding studies demonstrate that this decreased food intake accounts for previously described decreases in uterine and ovarian weights, which are much more affected by food restriction than is body weight. THC-treated rats lost weight initially which was not regained. Pair-fed rats gained only about one-half of the weight of the untreated control or vehicle-treated control rats over a 64-day period. Although long-term cannabinoid administration leads to tolerance and the resumption of the estrous cycle, the onset of estrus is often delayed when cannabinoid is administered 5-6 hr before the proestrus luteinizing hormone (LH) surge. Our results indicate that although chronic exposure to cannabinoids can continue to affect the rat estrous cycle, they do not have a direct effect on growth of the reproductive organs. The results reemphasize the need for adequate nutritional controls in marihuana and other toxicological research.

The effect of intragastric administration of delta 9-tetrahydrocannabinol on the growth and development of fetal mice of the A/J strain.

Pregnant A/J mice were intubated with vehicle (sesame oil:Tween 80:water) or 60, 120, or 240 mg/kg of delta 9-tetrahydrocannabinol on Days 11 and 12, 12 and 13, or 13 and 14 (vehicle and 240 mg doses only) of gestation. Mice were killed on Day 20 of gestation, and examined for number of corpora lutea and live and resorbed fetuses. Fetuses were weighed and examined for gross external and internal malformations. Each treatment group consisted of a minimum of 10 litters with about 10 pups per litter. In a few groups the effects of feed deprivation on Day 12 or of glucocorticoid administration on Days 12 and 13 (positive control) were assessed. Intubation with vehicle or delta 9-tetrahydrocannabinol, or feed deprivation did not affect number of live fetuses, incidence of resorption, fetal weights, or gross malformations other than cleft palate. Intubation of delta 9-tetrahydrocannabinol on gestational Days 12 and 13 or 13 and 14 increased the mean frequency of cleft palate formation. The increase was 2- to 2.5-fold at the 240-mg dose, being significant (p = 0.05) in the Days 12 and 13 group. Cortisone acetate and corticosterone injection induced both resorption and cleft palate formation. Other developmental or reproductive parameters were not influenced by delta 9-tetrahydrocannabinol treatment. We conclude that delta 9-tetrahydrocannabinol administered by gavage during Days 12 and 13 of gestation retards normal palatal development.

Effects of cannabinoids on function of testis and secondary sex organs in the Fischer rat.

Chronic oral treatment of young adult male Fischer rats with delta 9-tetrahydrocannabinol (THC), 1, 5 and 25 mg kg-1 day-1, or crude marihuana extract (CME), 3, 15 and 75 mg kg-1 day-1, reduced body weight gain by about 50-80% at the high CME or THC dose and was correlated with decreased food intake. When cannabinoid was administered early in the light cycle (9-11 a.m.), cauda epididymis sperm count and seminal vesicle fluid and fructose content were depressed to 50-65% at the high dosages but were not significantly different from those of pair-fed controls. Administration late in the light cycle (4-5 p.m.) depressed epididymal sperm count, seminal vesicle fluid content, and weight of testis, seminal vesicles and epididymis to 40-80% below that seen for pair-fed controls. 24 h after the last treatment, serum testosterone was unchanged in intubated control and low-dose treated rats, compared with untreated controls, but was elevated nearly twofold in medium-dose-treated rats (p less than 0.05). The results indicate that time of cannabinoid administration as well as feeding pattern are critical in studies of the rat reproductive system.

Effects of cannabinoids given orally and reduced appetite on the male rat reproductive system.

Chronic oral treatment of young adult male Fischer rats with delta 9-tetrahydrocannabinol (THC), 1, 5 and 25 mg/kg/day, or crude marihuana extract (CME), 3, 15 and 75 mg/kg/day, suppresses growth of accessory sex organs and body weight gain in a dose-related manner. Animals pair fed with the THC (25 mg/kg) group gained slightly more in body weight than the THC group, but their relative accessory sex organ weights were intermediate between THC and ad libitum-fed control group weights. These latter differences may be due to altered serum androgen levels since these levels 2-6 h after last treatment were 0.15, 0.77 and 3.33 ng/ml for THC, pair-fed and ad libitum-fed groups, respectively. 24 h after the last treatment all groups were within normal levels. Thus, chronic cannabinoid treatment suppresses accessory sex organ weights and serum androgen levels greater than the suppression caused by reduced food intake alone.

Effect of cannabinoids on estrous cycle, ovulation and reproductive capacity of female A/J mice.

Virgin A/J female mice were intubated daily for 8 days (short term) or 70 days (long term) with 0, 1, 5, or 25 mg/kg delta 9-tetrahydrocannabinol (delta 9-THC) or 0, 3, 15, or 75 mg/kg crude marihuana extract (CME) in a sesame oil:polysorbate 80:saline vehicle. These dosages approximate light, moderate, and heavy human usage. Short-term exposure to CME has no significant effect on PMS-HCG-induced ovulation but appears to: (1) delay entry into proestrus at all dose levels; (2) depress serum progesterone during the luteal phase at the highest CME level used (75 mg/kg), and (3) inhibit female receptivity to males at least at the highest dosage. Long-term oral administration of CME or delta 9-thc had no significant effect on length of estrous cycles or mating (plug formation) but term pregnancies were reduced by 32 and 68% for medium and high dosages, respectively. After a 30-day recovery period, 80% of those females that failed to have successful pregnancies now became pregnant.

Effect of colchicine on estrogen action. I. Inhibition of 17 beta-estradiol-induced water and potassium uptake in the immature rat uterus.

The effects of colchicine on 17 beta-estradiol-induced water and electrolyte uptake in the uterus of the immature rat have been examined 6 h after treatment with this estrogen. Estradiol stimulates an increase in total uterine Na+, K+ and water while intracellular Na+ and K+ concentrations remain relatively unchanged. Assuming the sodium space is equivalent to the extracellular space, the extracellular fluid compartment increases about 84% in response to estradiol. Similarly, the intracellular compartment increases by about 62%. The uptake of water into the cellular compartment may be a direct response to a stimulation of K+ accumulation by uterine cells. Colchicine inhibits both estradiol-induced rise in intracellular potassium and both intra- and extracellular water.

Effect of colchicine on estrogen action. II. Translocation of 17 beta-estradiol cytosol receptor complex into the nucleus.

Colchicine has previously been shown in our laboratory to inhibit 17 beta-estradiol stimulation of uterine water uptake in the immature rat measured 6 h after administration of the agents. We sought to determine whether this effect was mediated through colchicine action on translocation of estradiol receptor complex into the uterine cell nucleus. The time course of estradiol effect on uterine water uptake was followed with and without concurrent colchicine administration up to 6 h after administration. At no time during this period did there appear to be any influence of colchicine on translocation of the estradiol receptor complex into the nucleus. Examination of physical chemical characteristics of the nuclear estradiol receptr complex after estradiol and estradiol plus colchicine treatments revealed no observable differences. Thus, colchicine inhibition of estradiol-stimulated uterine water retention does not appear to be mediated through inhibition of nuclear translocation of estradiol-receptor complex nor to be due to any reduced retention time of estradiol-receptor complex in uterine nuclei.

Uterine tubulin production during early pregnancy in the rabbit.

Rabbit uterine tubulin levels rise rapidly in the preimplantation period to peak values of about 10-fold in the endometrium and about 4-5 fold in the myometrium on days 3-5 of pregnancy. Endometrial tubulin falls to non-pregnancy levels by mid-pregnancy while myometrial tubulin declines much more slowly. The rise in tubulin exceeds the early rate of increase in total uterine protein and DNA. Following implantation, tubulin content of the decidual basal plate area increases rapidly to peak about day 12 then declines. Production of uterine tubulin appears capable of a major, short term response to gonadal hormone stimulation.

Translational control of specific uterine protein synthesis after estrogen induction.

The rate of leucine incorporation into a specific estrogen-induced protein of immature rat isolated by gel electrophoresis declines rapidly between 2-4 hr after estrogen stimulation in vivo followed by incubation in vitro. Actinomycin D present during the in vitro phase prevents this decline, and elicits a "superinduction" effect at 2 hr. Labeled induced protein remains stable during this time period, indicating that its decline is due to a reduction in synthesis capacity for the inducible protein, rather than to its degradation. A second injection of hormone at 3 hr has no effect on the reduced level of synthesis capacity for induced protein noted at 4 hr in the rat uterus.