Optimization of warfarin dose by population-specific pharmacogenomic algorithm.

To optimize the warfarin dose, a population-specific pharmacogenomic algorithm was developed using multiple linear regression model with vitamin K intake and cytochrome P450 IIC polypeptide9 (CYP2C9(*)2 and (*)3), vitamin K epoxide reductase complex 1 (VKORC1(*)3, (*)4, D36Y and -1639 G>A) polymorphism profile of subjects who attained therapeutic international normalized ratio as predictors. New algorithm was validated by correlating with Wadelius, International Warfarin Pharmacogenetics Consortium and Gage algorithms; and with the therapeutic dose (r=0.64, P<0.0001). New algorithm was more accurate (Overall: 0.89 vs 0.51, warfarin resistant: 0.96 vs 0.77 and warfarin sensitive: 0.80 vs 0.24), more sensitive (0.87 vs 0.52) and specific (0.93 vs 0.50) compared with clinical data. It has significantly reduced the rate of overestimation (0.06 vs 0.50) and underestimation (0.13 vs 0.48). To conclude, this population-specific algorithm has greater clinical utility in optimizing the warfarin dose, thereby decreasing the adverse effects of suboptimal dose.

Evaluation of various parameters of calcium-alginate immobilization method for enhanced alkaline protease production by Bacillus licheniformis NCIM-2042 using statistical methods.

Calcium-alginate immobilization method for the production of alkaline protease by Bacillus licheniformis NCIM-2042 was optimized statistically. Four variables, such as sodium-alginate concentration, calcium chloride concentration, inoculum size and agitation speed were optimized by 2(4) full factorial central composite design and subsequent analysis and model validation by a second-order regression equation. Eleven carbon, 11 organic nitrogen and seven inorganic nitrogen sources were screened by two-level Plackett-Burman design for maximum alkaline protease production by using optimized immobilized conditions. The levels of four variables, such as Na-alginate 2.78%; CaCl(2), 2.15%; inoculum size, 8.10% and agitation, 139 rpm were found to be optimum for maximal production of protease. Glucose, soybean meal and ammonium sulfate were resulted in maximum protease production at 644 U/ml, 720 U/ml, and 806 U/ml when screened for carbon, organic nitrogen and inorganic nitrogen sources, respectively, using optimized immobilization conditions. Repeated fed batch mode of operation, using optimized immobilized conditions, resulted in continuous operation for 12 cycles without disintegration of beads. Cross-sectional scanning electron microscope images have shown the growth pattern of B. licheniformis in Ca-alginate immobilized beads.

Evaluation of two anesthetic techniques for hemodynamic control during carotid surgery. Preliminary results.

BACKGROUND: To evaluate two anesthetic techniques for hemodynamic control during carotid TEA surgery and early post-surgery. METHODS: Two study groups treated by carotid surgery were compared; the Fentanyl group consisted of 7 patients in ASA class 3, the Remifentanil-Sevoflorane group included 12 patients in ASA class 3. The double product was monitored on entry to the operating room, at 5, 15, 30 min after induction of anesthesia and tracheal intubation, and at 30 min after extubation. Time of extubation, re-awakening and attention levels during early post-surgery, and myocardial ischemia markers were monitored for 48 h after surgery in the Remifentanil group. RESULTS: Statistical analysis using Student's "t"-test for paired data showed that the double product indicated better hemodynamic stability in the patients who received Remifentanil-Sevoflorane than in those who received Fentanyl. CONCLUSIONS: Compared with anesthesia using Fentanyl and with locoregional techniques, anesthesia with Remifentanil-Sevoflorane in carotid surgery provides a valuable alternative and secures good hemodynamic stability.

An oral regimen of cyproterone acetate and testosterone undecanoate for spermatogenic suppression in men.

OBJECTIVE: To test the effectiveness, safety, and reversibility of the combined administration of cyproterone acetate and T undecanoate. DESIGN: Open clinical trial. SETTING: Healthy volunteers in an academic research environment. PATIENT(S): Eight healthy men, aged 25-42 years were selected. INTERVENTION(S): Cyproterone acetate, 12.5 mg, and T undecanoate, 80 mg, were administered orally twice daily for 16 weeks. MAIN OUTCOME MEASURE(S): Semen analyses every 2 weeks; physical examination, chemistries, hematology, prostatic-specific antigen, gonadotropins and T levels, and a questionnaire on sexual and behavioral function every 4 weeks. RESULT(S): In all subjects a profound suppression of spermatogenesis occurred; one subject became azoospermic, five subjects had sperm counts of < or = 3 x 10(6)/mL, and in two subjects sperm counts were 4 and 6 x 10(6)/mL in week 16. Sperm counts returned to baseline in all men after hormone administration was discontinued. No changes in metabolic parameters and total prostatic-specific antigen were detected. Hemoglobin and hematocrit decreased statistically significantly at week 16 of treatment and returned to baseline by week 12 of recovery. There was no change in sexual function or behavior. CONCLUSION(S): The oral administration of T undecanoate plus cyproterone acetate induces a profound suppression of spermatogenesis with no major adverse effects. These data suggest the feasibility of oral contraception in men.

A combined regimen of cyproterone acetate and testosterone enanthate as a potentially highly effective male contraceptive.

In this study we tested the effectiveness of the combined administration of cyproterone acetate (CPA) and testosterone enanthate (TE) in suppressing spermatogenesis. After a control phase of 3 months, 15 normal men were randomized to receive TE (100 mg/week) plus CPA at a dose of 100 mg/day (CPA-100; n = 5) or 50 mg/day (CPA-50; n = 5) or TE (100 mg/week) alone (n = 5) for 16 weeks. Semen analysis was performed every 2 weeks. Every 4 weeks, fasting blood samples were drawn for the measurement of LH, FSH, testosterone, estradiol, and biochemical and hematological parameters; subjects underwent a physical examination; and they and their partners filled in a sexual and behavioral questionnaire. Regardless of the dose, each of the 10 subjects receiving CPA plus TE became azoospermic, whereas only 3 of 5 subjects treated with TE alone achieved azoospermia. Times to azoospermia were 6.8 +/- 0.5, 8.4 +/- 1.0, and 14.0 +/- 1.2 weeks in groups CPA-100, CPA-50, and TE alone, respectively (P = NS). Throughout treatment, both gonadotropins tended to be higher in the TE alone group than in the other groups. This difference was mostly due to the higher gonadotropin levels present in the 2 men treated with TE alone that remained oligospermic. No difference in testosterone or estradiol levels was found among the groups. No significant change in lipoprotein levels or liver function tests could be detected. In the CPA-100 and CPA-50 groups, hemoglobin, hematocrit, and red blood cells were lower at the end of the treatment phase, whereas no change was detected in TE alone group. A tendency for a decrease in body weight was detected in subjects treated with CPA, whereas there was no change in subjects receiving TE alone. At the end of the treatment phase, a decrease in testis size was present in all groups. There was no significant change in sexual function, aggressive behavior, mood states, or satisfaction with relationship in any group. These results suggest that the combined administration of CPA and TE is very effective in suppressing spermatogenesis and may represent a promising regimen for reversible contraception in males.

Different gonadotropin and leuprorelin ovulation induction regimens markedly affect follicular fluid hormone levels and folliculogenesis.

OBJECTIVES: To clarify the endocrine mechanisms underlying the outcome of different ovulation induction regimens with gonadotropins and GnRH agonists (GnRH-a). DESIGN: Prospective study. SETTING: Reproductive Endocrinology Center, University of Bologna. PATIENTS: Forty eumenorrheic women randomly assigned to four groups of 10 subjects each. INTERVENTIONS: Ovulation induction regimens: group A, purified FSH only; group B, purified FSH and flare-up GnRH-a; group C, purified FSH and long GnRH-a; and group D, hMG and long GnRH-a. MAIN OUTCOME MEASURES: Pelvic ultrasound and hormone levels in daily serum samples and in follicular fluid drawn immediately before hCG administration. RESULTS: Exogenous gonadotropin dose did not differ among groups. Group B had fewer preovulatory follicles than group C. Group B had higher serum LH, FSH, E2, P, T, and follicular fluid LH, E2, T, and alpha-inhibin than groups C and/or D. Groups C and D did not differ. CONCLUSIONS: Long GnRH-a regimens improved follicle yield and the endocrine milieu in spite of comparable exogenous gonadotropin dose and lower serum FSH and thus appear to be preferable in assisted reproduction. Reduced folliculogenesis found in flare-up GnRH-a regimens could be mediated by the atretic effects of high intraovarian androgens. Efficacy of purified FSH and hMG was comparable.

Dissociated 24-hour patterns of somatotropin and prolactin in fatal familial insomnia.

To assess the changes in the 24-hour profiles of serum somatotropin and prolactin levels during total disruption of the sleep/wake cycle sustained over several months, we studied 2 subjects affected by fatal familial insomnia, a rare disease characterized by selective thalamic degeneration that causes chronic sleep loss. Under standardized conditions and polysomnographic control, the patients underwent repeated 24-hour study sessions covering the entire clinical course of the disease. Hormones were assayed at 30-min intervals. Four healthy volunteers were used as controls. A sleep/wake cycle was always absent in fatal familial insomnia. Serum somatotropin and prolactin concentrations never exceeded the normal range of variation. The nocturnal elevation of somatotropin disappeared simultaneously with sleep loss, whereas a significant 24-hour component of variations in serum prolactin levels was present for months after total disruption of the sleep/wake cycle, with normally placed nocturnal acrophases. Complete obliteration of the 24-hour component was achieved for prolactin only in the advanced stages, through a progressive decrease in 24-hour amplitude of variation. Selective and progressive degeneration of the mediodorsal and anterior ventral nuclei of the thalamus causes an early obliteration of the 24-hour rhythm of somatotropin and a later disappearance of circadian prolactin rhythmicity.(ABSTRACT TRUNCATED AT 250 WORDS)

Diurnal blood pressure variation and hormonal correlates in fatal familial insomnia.

Fatal familial insomnia is a prion disease in which a selective thalamic degeneration leads to total sleep deprivation, hypertension, dysautonomia, adrenal overactivity, and impaired motor functions. With patients under continuous recumbency and polysomnographic control, we assessed the changes in the 24-hour patterns of blood pressure, heart rate, plasma catecholamines, corticotropin, and serum cortisol in three patients at different stages of the disease. Six healthy volunteers were used as control subjects. A dominant 24-hour component was detected at rhythm analysis of all variables, both in patients and control subjects. In the patients, the amplitudes gradually decreased as the disease progressed, leading to the obliteration of any significant dirunal variation only in the preterminal stage. A shift in phase corresponded to the loss of the nocturnal fall in blood pressure in an early stage of the disease, when nocturnal bradycardia was still preserved. Plasma cortisol was high and became increasingly elevated, whereas corticotropin remained within normal levels; abnormal nocturnal peaks appeared in their circadian patterns. The disrupted patterns of cortisol and blood pressure preceded the development of hypertension and severe dysautonomia, which in turn were paralleled by increasing catecholamine and heart rate levels. Our data demonstrate that in patients with fatal familial insomnia the changes detectable in the rhythmic component of diurnal blood pressure variability result in a pattern of secondary hypertension. Disturbances in thalamic, pituitary-adrenal, and autonomic functions seem to be involved in mediating these changes.

Progressive disruption of the circadian rhythm of melatonin in fatal familial insomnia.

Fatal familial insomnia (FFI) is a disease characterized by loss of sleep activity due to selective thalamic degeneration. To assess the secretory pattern of melatonin (MT) in FFI, we studied two cases of overt disease under standardized conditions and polysomnographic control. Each patient underwent repeated 24-h study sessions, and MT was assayed at 30-min intervals. Six healthy volunteers were used as controls. Slow wave sleep was never recorded, whereas occasional episodes of enacted dreaming accompanied by rapid ocular movements and complex muscular activities were documented, with no detectable rhythm. Plasma MT concentrations gradually decreased as the disease progressed. A significant circadian rhythm was detected in the earlier recordings, with decreasing amplitudes with disease progression. Complete rhythm obliteration was achieved in the most advanced stage. Normally placed nocturnal acrophases were detected in the earlier stages, but then a shift toward the daytime hours was observed. Thalamic lesions of FFI appear to determine a progressive disruption of the sleep/wake cycle accompanied by decreased circulating levels of MT, with progressive alterations in the circadian rhythm of this hormone. On the other hand, decreased secretion of MT may contribute to the sleep disturbances of FFI.

Accuracy of transvaginal ultrasound and serum hCG in the diagnosis of ectopic pregnancy.

Transvaginal ultrasound was performed upon admission of 127 patients with a clinical suspicion of ectopic pregnancy in association with human chorionic gonadotropin (hCG) determination. Failure to visualize with sonography an intrauterine gestational sac with an hCG level superior to 1000 mIU/ml identified 25/42 tubal pregnancies with a positive predictive value of 86% and a specificity of 93%. Abnormal adnexal findings occurred in 95% of the ectopic pregnancies. Extrauterine gestational sacs with or without embryos could be confidently detected in 19 ectopic pregnancies (45%). A complex adnexal mass was seen in 19 cases and yielded a positive predictive value of 90% (19/21). Adnexal gestational sacs and complex masses were seen more frequently in those ectopic pregnancies with an hCG level above 1000 mIU/ml but the difference was not significant (100% versus 78%). Simple adnexal cysts were found more frequently in intrauterine pregnancies, and fluid in the cul-de-sac was also not indicative of ectopic pregnancy (positive predictive value, 29%). Transvaginal ultrasound has a primary role in the diagnosis of ectopic pregnancy. The combined use of uterine and adnexal sonography associated with elevated hCG levels allows a definitive diagnosis in the vast majority of cases at a very early stage, when the chances for a successful conservative treatment are greater.

Circadian hormonal rhythms in two new cases of fatal familial insomnia.

We used a chronobiological inferential statistical method to investigate circadian rhythms of hypophyseal hormones, cortisol, melatonin and catecholamines in two females of the same family affected by fatal familial insomnia. Case 1 (confirmed at autopsy) presented an absent or progressive loss of circadian rhythms of all hormones. In case 2 there was a loss of GH circadian rhythm and a less significant rhythm for melatonin, catecholamines and gonadotropins. These results confirm the role of the thalamus in regulating hormonal circadian rhythm.

[Anesthesia and sickle cell disease]. / Anestesia e drepanocitosi.

Sickle cell anaemia is the most common hereditary haemoglobin pathology. It is found in either a homozygous or heterozygous form, associated in the latter case with other haemoglobinopathies. In view of the pathogenesis and the various related imbalances, amply confirmed by others, which can well prove disastrous, the pre, per and post-operative precautions to be adopted in such patients are assessed.