Inhibition by an ergoline derivative (dopaminergic agonist) of oestradiol-induced adenohypophyseal growth and of the decrease in the hypothalamic ascorbic acid (HAA) concentration in rats

The increase in adenohypophyseal weight and the decrease in the ascorbic acid concentration in the hypothalamus of rats injected i.m. twice a week with oestradiol benzoate as an aqueous microcrystal suspension in doses of 2.65 mumol (1 mg) was completely (HAA) or almost completely (adenohypophyseal weight) inhibited by the simultaneous administration of the ergoline derivative D-6-methyl-8-ergoline-(1)-yl acetic acid amide (Deprenon SPOFA) in the diet in daily doses of 0.28 mumol (200 micrograms) per rat. The functional significance of HAA in dopaminergic modulation of oestrogen-induced adenohypophyseal growth is discussed, with special reference to the possible function of HAA as a dopamine-beta-hydroxylase cofactor.     

The effect of hexose monophosphate shunt inhibitors on adenohypophyseal and ceruloplasmin reactions to oestrogen.

Oestradiol benzoate, as an aqueous microcrystal suspension, was administered i.m. to rats in doses of 1 mg twice a week; it induced adenohypophyseal hyperplasia and an increase of the thyroxine-binding capacity of the adenohypophyseal proteins in vitro and raised the blood ceruloplasmin level. The simultaneous administration of a hexose monophosphate shunt inhibitor--6-aminonicotinamide (200 microgram/rat/day in food) or oxythiamine (8 mg/rat/day in food)--did not modify the reaction of the adenohypophysis; the hexose monophosphate shunt thus probably does not play a significant role in the adenohypophyseal reaction to oestrogens. By themselves, both inhibitors raised the blood ceruloplasmin level and their effect summated with that of oestradiol. The mechanism of action of the inhibitors is not known, but a nonspecific stress effect leading to an increase in the ceruloplasmin level as an "acute phase protein" is considered to be the most likely.     

Metoclopramide potentiates the hypophyseal reactions to oestradiol

Male rats to which oestradiol benzoate was administered intramuscularly twice a week for three weeks in 1 mg doses as an aqueous microcrystal suspension showed an increase in adenohypophyseal weight, in the number of lactotropic cells in the adenohypophysis (demonstrated by immunohistochemical detection of prolactin) and in polyphenol oxidase (ceruloplasmin) activity in the blood and hypothalamus. The simultaneous administration of metoclopramide (methoxychloroprocainamide) in doses of 10 mg/rat per day in food potentiated the adenohypophyseal reaction to oestradiol (weight and the number of lactotropic cells), but potentiated the polyphenol oxidase reaction only little or not at all. Metoclopramide thus has an anti-dopaminergic effect similar to that of perphenazine and other inhibitors of dopaminergic neurones.     

Dopaminergic antagonists potentiate the adenohypophyseal growth reaction to oestrogen: thioridazine is less effective than perphenazine

The injection of oestradiol benzoate as an aqueous microcrystal suspension, in a dose of 1 mg twice a week, evokes a marked adenohypophyseal growth reaction in male rats. The reaction is potentiated by dopaminergic antagonists from the group of neuroleptics (specifically perphenazine and thioridazine). Elicitation of the same adenohypophyseal reaction required twice as much thioridazine (10 mg/rat per day) as perphenazine. Thyroxine inhibited the adenohypophyseal growth reaction to oestradiol. The serum polyphenol oxidase (ceruloplasmin) level rose after oestradiol and perphenazine and thioridazine slightly potentiated the increase.     

Inhibition of thyroxine binding to adenohypophysial proteins in vitro by 2,4-dinitrophenol administered in vivo.

Male and female rats were given oestradiol benzoate (1 mg as a microcrystal aqueous suspension i.m. twice a week), 0.0033% 2.4-dinitrophenol (DNP) in their food (about 1 mg/rat/day), or 0.1% DNP in their food (about 30 mg/rat/day), or both oestradiol and DNP. The smaller DNP dose mildly stimulated food consumption and did not affect body weight. The larger dose strongly inhibited food consumption in the first two weeks of the experiment; consumption then returned to the control level, but body weight fell markedly at the same time. After 3 weeks' administration of both the small and the large dose of DNP, adrenal weight in the males was raised and the weight of the gonads was unchanged. The large DNP dose severely reduced the weight of the seminal vesicles and the uteri. It also inhibited the accumulation of radioiodine in the thyroid of both males and females. Isolated administration of the oestrogen raised adrenal weight in the males and ovarian and uterine weight in the females; it reduced the weight of the testes and seminal vesicles. These reactions were not affected by DNP. A pronounced oestradiol-induced increase in the weight of the adenohypophyses was accompanied by raised thyroxine binding to the adenohypophysial proteins in vitro. DNP inhibited the growth reaction of the adenohypophysis to the oestrogen only slightly and non-significantly, but significantly inhibited the thyroxine binding reaction to the adenohypophysial proteins in vitro. By itself, DNP had no effect on adenohypophysial weight, but reduced thyroxine binding to the adenohypophysial proteins in vitro, especially in males. The effect of DNP was similar to that of thyroxine observed in earlier experiments; nothing is known of its mechanism.     

Interaction of perphenazine and an ergoline derivative on oestrogen-induced adenohypophyseal growth.

Male and female rats were injected twice a week for three weeks with doses of 1 mg oestradiol benzoate (OE), were given perphenazine (P, 2 mg/rat/day) or the ergoline derivative D-6-methyl-8-ergoline-(I)-yl acetic acid amide (Deprenon SPOFA, D, 200 microng/rat/day) in their food or were treated with various combinations of all three factors. OE-induced adenohypophyseal growth was inhibited by D, but the inhibitory effect of D was completely suppressed by P. D also inhibited the OE-induced increase in the thyroxine-binding capacity of the adenohypophyseal proteins, but this inhibition was not suppressed by the simultaneous administration of P. The administration of OE was followed by elevation of the serum ceruloplasmin level, which was not inhibited by P or D, either alone or combined. Ovarian weight rose markedly after D and the increase was inhibited by the simultaneous administration of either OE or P.     

Modulation of the effect of oestradiol on adenohypophyseal weight: ineffectiveness of nickel chloride, potentiation by cimetidine

Male rats received an i.m. injection of oestradiol benzonate twice a weak, as an aqueous microcrystal suspension in doses of 1 mg, and/or were given, in their food, nickel chloride in daily doses of 10 mg [first experiment] or 20 mg [second experiment] per rat, or cimetidine, an antagonist of H2 receptors, in daily doses of 20 mg [first experiment] or 40 mg [second experiment] per rat. Neither dose of nickel chloride affected the oestrogen-induced growth reaction of the adenohypophysis, the increase in polyphenol oxidase [ceruloplasmin] activity in the blood [the larger dose stimulated it slightly], the increase in hypothalamic polyphenol oxidase activity or the post-oestrogen drop in the hypothalamic ascorbic acid concentration. Both doses of cimetidine potentiated the growth reaction of the adenohypophysis to oestrogens, but did not affect the blood polyphenol oxidase, the hypothalamic polyphenol oxidase or the hypothalamic ascorbic acid reaction. If administered alone, the larger dose of cimetidine mildly reduced serum polyphenol oxidase [ceruloplasmin] activity.     

Dopaminergic control of adenohypophyseal growth after oestrogens: negative results with L-DOPA, RO-4-4602, alpha-methyl-DOPA, apomorphine, haloperidol, pyridoxine and cyproheptadine.

Chronic oestrogen treatment augments adenohypophyseal weight and the thyroxine-binding capacity of adenohypophyseal proteins in rats. Since these reactions are both inhibited by ergocornine and ergoline derivatives (as well as by the thyroid hormones, testosterone, anti-oestrogens and antiandrogens), and are potentiation by perphenazine and the dopaminergic neurone blocker Pimozide, the peroral effectiveness of various other substances presumed to act in the region of the dopaminergic or serotoninergic neurones of the hypothalamus was tested. L-DOPA (10 mg/rat/day) did not modify the adenohypophyseal reaction, either alone or combined with the DOPA-decarboxylate inhibitor Ro-4-4602 (1 mg/rat/day). alpha-Methyl-DOPA (10 mg/rat/day), apomorphine (3 mg/rat/day), haloperidol (0.2 mg/rat/day), pyridoxine (20 mg/rat/day) and cyproheptadine (1 mg/rat/day) were likewise ineffective.     

Antioestrogenic action of the aldosterone antagonist canrenoate K in the rat (adenohypophysis, ceruloplasmin).

In a dose of 7,k mg/rat/day in food, the aldosterone antagonist canrenoate K inhibited the adenohypophyseal reaction (growth, raised thyroxine-binding capacity of the adenohypophyseal proteins in vitro) and the ceruloplasmin reaction (elevation of the serum ceruloplasmin level) to three weeks' intramuscular administration of long-acting oestradiol benzoate in doses of 1 mg twice a week. The effect was similar to that of the antioestrogen clomiphen in a dose of 1.25 mg/rat/day in food. In combined administration of clomiphen and canrenoate K, no summation of their effect was observed. Neither canrenoate nor clomiphen affected the post-oestradiol drop in body weight, but they both potentiated the oestradiol-induced decrease in testicular weight and canrenoate potentiated the effect of oestradiol on uterine weight. It was therefore concluded that the effect of canrenoate is not of a catatoxic nature, i.e. that it is not determined by increased metabolic degradation of oestradiol.     

Thyroxine cardiomegaly in rats is not hypertension-bound

For 13-27 days, male rats were given dried thyroid (Thyreoidin SPOFA) in their food, 0.9% NaCl to drink instead of water, or both. In all the experiments, Thyreoidin induced pronounced cardiomegaly. The blood pressure rose only after 0.9% NaCl combined with Thyreoidin and only in experiments lasting 13 or 16 days (not 25 and 27 days). After three weeks of the experiment, therefore, hypertension disappeared, but cardiomegaly persisted. In all the experimental groups, including the controls, a significant positive correlation was found between heart weight and adrenal weight, but not between heart weight and blood pressure or between adrenal weight and blood pressure. The adrenals may thus participate in some way in the development of thyroxine cardiomegaly.     

Reactions of hypothalamic ascorbic acid, serum ceruloplasmin and the adenohypophysis to oestradiol: inhibition by L-thyroxine

Four weeks' administration of oestradiol benzoate to male and female rats in doses of 1 mg twice a week leads to adenohypophyseal hyperplasia and to an increase in the thyroxine-binding capacity of the adenohypophyseal proteins in vitro. At the same time, serum polyphenol oxidase (ceruloplasmin) activity rises and the hypothalamic ascorbic acid concentration falls. The simultaneous administration of L-thyroxine (0.1 mg/rat/per day) or dried thyroid (but not D-thyroxine) significantly inhibits these changes (adenohypophysis, ceruloplasmin) or completely suppresses them (hypothalamic ascorbic acid). L-thyroxine evidently blocks the action of oestradiol in the adenohypophysis, the liver and the hypothalamus; the significance of this inhibition is discussed in relation to dopaminergic modulation of the adenohypophyseal reaction to oestradiol.     

Interferon-beta (INF-beta) antibodies in interferon-beta1a- and interferon-beta1b-treated multiple sclerosis patients. Prevalence, kinetics, cross-reactivity, and factors enhancing interferon-beta immunogenicity in vivo

We analysed the role of dosage, route and frequency of administration of clinical grade interferon-beta (IFN-beta) preparations in inducing anti-IFN-beta antibodies (IFN-beta-Abs) in 5 groups of relapsing-remitting multiple sclerosis (RRMS) patients who were respectively treated as follows: 1) weekly intramuscular (i.m.) injections of 30 mg of recombinant IFN-beta1a (Avonex), 2) subcutis (s.c.) injections of 250 mg IFN-beta1b (Betaferon) every other day, 3) weekly i.m. injections of 250 mg IFN-beta1b (Betaferon), 4) s.c. injections of 22 mg of IFN-beta1a (Rebif) three times a week, and 5) i.m. injections of 22 mg of IFN-beta1a (Rebif) twice a week. IFN-beta-Abs were determined by ELISA. IFN-beta1b was more immunogenic than IFN-beta1a not only when administered s.c. every other day, but also when administered i.m. at a lower weekly dose; i.m. injection, however, significantly delayed the appearance, and induced lower serum levels of IFN-beta-Abs. In patients treated with s.c. IFN-beta1b, Ab levels peaked 3 to 9 months after therapy initiation, and then slowly, but progressively, declined to pre-therapy levels that in some patients were reached after three years. Patients treated with i.m. or s.c. IFN-beta1a only rarely developed IFN-beta-Abs, and then at very low titers. Overall, the i.m. weekly administration of IFN-beta1a was the less immunogenic treatment. In IFN-beta1b-treated patients, a wash-out period of two/three months was sufficient to bring the IFN-beta-Ab levels below the cut-off. Our findings suggest that the immunogenicity of IFN-beta1a is low, regardless of the route of administration and the dosage, while that of IFN-beta1b is high, and is significantly, but not completely reduced by i.m. administration. As IFN-beta-Abs are cross-reactive, a wash-out period is suggested when the preparation is changed from IFN-beta1b to IFN-beta1a in order to maintain the clinical benefits of the therapy.     

Characterization of Streptozotocin-induced Diabetic Rats and Pharmacodynamics of Insulin Formulations

Morphological and functional changes of rat pancreatic islets caused by administration of streptozotocin (STZ) and the bioavailability of insulin formulations administered to STZ-induced diabetic rats with fasting (12 h) or non-fasting were investigated. Islets isolated from normal rats maintained a good three-dimensional structure and the islet yield was 962.5±86.5 islet equivalent number (IEQ, islets converted to an average diameter of 150 μm). In the diabetic group (>500 mg/ml blood glucose), the islet yield was only 44.4±8.3 IEQ and the islet was severely damaged. The minimum reduction of blood glucose of each formulation, such as insulin solution, microcrystal, and insulin microcrystal capsule, was shown to be 11.3, 11.0, and 16.3 mg/dl, respectively, at 6 h in fasting with diabetic rats. These results indicated that the administration of insulin formulations to the fasting groups increased the severe hypoglycemic effect of insulin action more than in non-fasting diabetic rats. The diabetic rat with fasting has a regulatory disorder in maintaining the blood glucose level. Accordingly, the validity of pharmacological availability as an optimal modeling of insulin formulations is best in non-fasting STZ-induced diabetic rats.     

Prevention of N-methylnitrosourea-induced colon carcinogenesis in rats by oxygenated carotenoid capsanthin and capsanthin-rich paprika juice

Epidemiological and animal studies have provided evidence that dietary carotenoids may reduce the risk of certain types of cancer. An inhibitory activity of oxygenated carotenoid capsanthin, a potent antioxidant, and paprika juice rich in capsanthin (3.54 mg/100 ml) against colon carcinogenesis was investigated in F344 rats. In Experiment I (short-term assay), six rats each were given a gavage of 5 mg, 0.2 mg, or 0.008 mg capsanthin six times a week for Weeks 2-6 after receiving three intrarectal doses of 4 mg N-methylnitrosourea in Week 1. The number of colonic aberrant crypt foci, preneoplastic lesions, at Week 6 was significantly fewer (by 42%) in the 0.2 mg capsanthin group, but not in other groups, than the control group. In Experiment II (long-term assay), five groups of 30 or 25 rats each received an intrarectal dose of 2 mg N-methylnitrosourea three times a week for Weeks 1-3, and had either of 10 p.p.m. or 2 p.p.m. capsanthin solutions, 1:2.5 and 1:16.7 diluted solution of paprika juice (containing 10 p.p.m. or 2 p.p.m. capsanthin), and tap water (control fluid) as drinking fluid throughout the experiment. The experimental groups were fed 0.2 mg or 0.04 mg capsanthin/day/rat. The colon cancer incidence at Week 30 was significantly lower in the highly diluted paprika juice group (40%), but not in the moderately diluted paprika juice group (60%) and the capsanthin solution groups (68% and 68%) than the control group (83%). The results suggested that paprika juice may affect colon carcinogenesis. However, capsanthin alone failed to inhibit colon tumorigenesis, in spite of suppression of aberrant crypt foci formation in the short-term assay. Further studies are needed to explain this discrepancy.     

Adverse reactions to frusemide in hospital inpatients.

Out of 2580 medical inpatients included in a drug-surveillance programme, 585 (22.7%) were treated with frusemide. Of these, 123 (21.0%) had a total of 177 adverse reactions. The most common were hypovolaemia (85 cases), hyperuricaemia (54), and hypokalaemia (21). Most reactions were mild, and only three patients had potentially life-threatening effects. The incidence of adverse reactions increased significantly with daily dose, occurring in 47 patients (13.5%) given up to 40 mg, 42 (26.3%) given up to 80 mg, and 34 (43.6%) given over 80 mg (P less than 0.001). There was no clear association between side effects and a raised blood urea concentration on admission, confirming that treatment with frusemide is not more hazardous in patients with renal failure. Frusemide is a safe and highly effective diuretic. Nevertheless, in view of the potential seriousness of volume depletion, dosage should probably begin at 20 rather than 40 mg daily.     

布地奈德混悬液雾化吸入辅助治疗婴幼儿毛细支气管炎的最佳剂量研究

目的:探讨不同剂量布地奈德(Budesonide,BUD)混悬液雾化吸入对于婴幼儿毛细支气管炎进行治疗的临床效果。方法:选取我院2013年6月-2014年6月间收治的120例毛细支气管炎患儿为研究对象,采用随机数字表法将患儿随机分为高剂量组(44例)、低剂量(40例)和对照组(36例)三组。对照组进行综合治疗,高剂量组和低剂量组在综合性治疗的基础上分别给予BUD混悬液1 mg/次与0.5 mg/次治疗。观察三组患儿不良反应的发生情况,并比较他们的气促缓解时间、咳嗽消失时间、哮鸣音消失时间以及住院时间是否存在差异。结果:高剂量组的气促缓解时间、咳嗽消失时间、哮鸣音消失时间以及住院时间均明显低于低剂量组和对照组,差异均有统计学意义(P0.05);低剂量组的气促缓解时间、咳嗽消失时间、哮鸣音消失时间以及住院时间明显低于对照组,差异均有统计学意义(P0.05)。高剂量组出现3例鹅口疮(6.82%),低剂量组出现2例(5.00%),两组比较,差异无统计学意义(x2=0.124,P=0.725),且经对症处理后很快治愈,三组未见其他不良反应。结论:高剂量BUD混悬液雾化吸入佐治患儿毛细支气管炎具有疗效好,成本低和安全性高的特点,但临床应注意患者不良反应的发生。     

Human Insulin Microcrystals with Lactose Carriers for Pulmonary Delivery

Dry powder formulations for pulmonary delivery are attractive because many issues of solubility and stability can be minimized. Human insulin microcrystals with lactose carriers were produced for pulmonary delivery. The average particle diameter was 2.3 μm, with a narrow, monodispersed size distribution. The percentages of high molecular weight proteins (%HMWPs), other insulin-related compounds (%OIRCs), and A-21 desamido insulin (%D_es) were very low throughout the microcrystal preparation process. Administration of the microcrystal powder by intratracheal insufflation significantly reduced the blood glucose levels of Sprague-Dawley rats. The percent minimum reductions of the blood glucose concentration (%MRBG) produced by the insulin microcrystal powder and by an insulin solution reached 40.4% and 33.4% of the initial glucose levels respectively, and their bioavailability relative to subcutaneous injection (F) was 15% and 10% respectively. These results confirm that the insulin microcrystal powder prepared is suitable for pulmonary delivery in an effective dosage form.     

Evaluation of three estrus synchronization regimens for use in extensively managed Bos indicus and Bos indicus/taurus heifers in Northern Australia

Two herds of extensively managed yearling Bos indicus and Bos indicus x Bos taurus heifers (n = 169) were assigned to one of three estrus synchronization treatment groups. The treatments consisted of 1) two injections of the prostaglandin (F(2)alpha) analogue, luprostiol (7.5 mg), given 12-d apart; 2) insertion of an intravaginal progesterone-releasing device (CIDR-B), followed by 400 IU i.m. pregnant mare serum gonadotrophin (PMSG) and 7.5 mg luprostinol at CIDR-B removal or 3) subcutaneous progestagen implant (SMB) for 10-d, followed by 400 IU i.m. and 7.5 mg i.m. luprostiol at implant removal. A dual tail paint-raddle system was used to evaluate response to treatment. Each treatment group was inseminated according to the manufacturer's recommendations for fixed-time artificial insemination (A.I.; luprostiol, 72h after the second injection, SMB and CIDR-B 48h after implant/device removal). The overall 51-d pregnancy rates for the three treatments were 18.6% (luprostiol), 48.8% (CIDR-B), and 53.3% (SMB). There were significant differences in the pregnancy rate between CIDR-B and luprostiol (P = 0.004) and between SMB and luprostiol (P =< 0.0001), but there was no difference in pregnancy rate between CIDR-B and SMB treatment.     

Gemcitabine and irinotecan as first-line therapy for carcinoma of unknown primary: results of a multicenter phase II trial

Metastatic carcinoma of unknown primary (CUP) has a very poor prognosis, and no standard first-line therapy currently exists. Here, we report the results of a phase II study utilizing a combination of gemcitabine and irinotecan as first-line therapy. Treatment was with gemcitabine 1000 mg/m(2) and irinotecan 75 mg/m(2) weekly times four on a six week cycle (Cohort I). Due to excessive toxicity, the dose and schedule were modified as follows: gemcitabine 750 mg/m(2) and irinotecan 75 mg/m(2) given weekly times three on a four week cycle (Cohort II). The primary endpoint was the confirmed response rate (CR + PR). Secondary endpoints consisted of adverse events based upon the presence or absence of the UDP glucuronosyltransferase 1 family, polypeptide A1*28 (UGT1A1*28) polymorphism, time to progression, and overall survival. Thirty-one patients were enrolled with a median age of 63 (range: 38-94), and 26 patients were evaluable for efficacy. Significant toxicity was observed in Cohort 1, characterized by 50% (7/14) patients experiencing a grade 4+ adverse event, but not in cohort II. The confirmed response rate including patients from both cohorts was 12% (95% CI: 2-30%), which did not meet the criteria for continued enrollment. Overall median survival was 7.2 months (95% CI: 4.0 to 11.6) for the entire cohort but notably longer in cohort II than in cohort I (9.3 months (95% CI: 4.1 to 12.1) versus 4.0 months (95% CI: 2.2 to 15.6)). Gemcitabine and irinotecan is not an active combination when used as first line therapy in patients with metastatic carcinoma of unknown primary. Efforts into developing novel diagnostic and therapeutic approaches remain important for improving the outlook for this heterogeneous group of patients. TRIAL REGISTRATION: ClinicalTrials.gov NCT00066781.     

Establishment of a system of high-frequency embryogenesis from long-term cell suspension cultures of rice (Oryza sativa L.)

Summary Suspension cultures which maintained embryogenic potency for more than 18 months were established from excised immature embryos of rice (Oryza sativa L. cv. Konansou). The cultures were subcultured every three days in N6 medium supplemented with proline (10 mM), casein hydrolysate (300 mg/l), sucrose (30 g/l) and 2,4-D (1 mg/l). The frequency of embryogenesis from the embryogenetic suspension cultures reached about 90% when cell clusters (about 1 mm in diameter) were transferred to a solid medium which consisted of N6 medium, NAA (1 mg/l), kinetin (5 mg/l), sucrose (30 g/l) and Gelrite (2 g/l). When smaller clusters of cells (approximately 200–400 m in diameter) were transferred to a liquid medium which consisted of salts of N6 medium diluted with an equal volume of water plus sucrose (45 g/l), NAA (0.01 mg/l) and 4-PU (0.1 mg/l) at a cell density of 13 clusters/ml in 2 ml of medium, somatic embryogenesis was initated at high frequency (about 50%). Morphological evidence is provided to demonstrate that the regeneration occurred via embryogenesis. This is the first report of high-frequency embryogenesis in suspension cultures of rice cells.