Phase I and pharmacokinetic studies of PNU-159548, a novel alkycycline, administered intravenously to patients with advanced solid tumours.

PNU-159548 (4-demethoxy-3'-deamino-3'-aziridinyl-4'-methylsulphonyl-daunorubicin) is the lead compound of a novel class of cytotoxic agents (alkycyclines) with a unique mechanism of action combining DNA intercalation with alkylation of guanines in the DNA major groove. The objectives of two phase I studies were to assess the dose-limiting toxicities (DLTs), to determine the maximum tolerated dose (MTD) and to study the pharmacokinetics (PKs) of PNU-159548 and its active metabolite PNU-169884 when administered intravenously (i.v.) over 10 or 60 min to patients with advanced solid tumours. Patients were treated with escalating doses of PNU-159548, courses repeated every 21 days at doses ranging from 1.0 to 16 mg/m(2). For pharmacokinetic analysis, plasma sampling was performed during the first course and assayed using a validated high-performance liquid chromatographic assay with mass spectrometric detection. 69 patients received a total of 161 courses. The MTD was reached at 14 and 16 mg/m(2) in heavily (HP) and minimally pretreated/non-pretreated (MP) patients, respectively, with thrombocytopenia as the DLT. A hypersensitivity reaction was observed in 8 patients across all dose levels, characterised by fever with chills, erythema, facial oedema and dyspnoea. The PKs of PNU-159548 and PNU-169884 were linear over the dose range studied. A significant correlation was observed between the percentage decrease in platelet count and the AUC of PNU-159548. In these studies, the DLT of PNU-159548 was thrombocytopenia. The recommended dose for phase II studies of PNU-159548 is 12 and 14 mg/m(2) administered i.v. over 10 min, once every 21 days, in HP and MP patients, respectively.

The impact of drug administration sequence and pharmacokinetic interaction in a phase I study of the combination of docetaxel and gemcitabine in patients with advanced solid tumors.

Our objective was to determine the maximum tolerated dose (MTD) of two administration sequences of docetaxel and gemcitabine in cancer patients, and to describe the pharmacokinetics of both drugs. Patients were treated in a 4-weekly schedule at two dose levels: gemcitabine 800 mg/m2 on days 1, 8 and 15, and docetaxel 85 or 100 mg/m2 on day 15 (levels I and II). The protocol was amended to a 3-weekly schedule, testing gemcitabine 800 or 1000 mg/m2 on days 1 and 8, with docetaxel 85 mg/m2 on day 8 given initially (dose levels IIIa and IV). At the recommended dose, an extra cohort of patients initially received gemcitabine (dose level IIIb). Eleven patients were treated with the 4-week schedule; 29% of cycles were delayed predominantly because of hematological toxicity. Four patients developed dose-limiting toxicities (DLTs), predominantly hematological. In the 3-week schedule, 14 patients were treated. At level IV, three of four patients developed DLTs, defining the MTD. With the reverse sequence, three patients received a total of 10 cycles. Overall, nine partial remissions were observed. We conclude the recommended dose for phase II studies is gemcitabine 800 mg/m2 on days 1 and 8, combined with docetaxel 85 mg/m2 on day 8, on a 3-weekly schedule. Gemcitabine distribution is significantly altered upon docetaxel administration.

Dose and schedule-finding study of oral topotecan and weekly cisplatin in patients with recurrent ovarian cancer.

Both weekly cisplatin chemotherapy and single agent topotecan have proven to be effective in recurrent ovarian cancer. Preclinical data show synergism between cisplatin and topotecan. Side effects for this combination are drug sequence dependent and predominantly haematologic. Since preclinical data suggest that Cremophor EL (CrEL), the formulation vehicle of paclitaxel, has a protective effect on haematological toxicity of cisplatin, CrEL was added to the combination cisplatin and topotecan. In this phase I study, escalating doses of oral topotecan administered on day 1, 2, 8, 9, 15, 16, 29, 30, 36, 37, 43, 44 were combined with weekly cisplatin 70 mg m(-2) d(-1) on day 1, 8, 15, 29, 36, 43 (scheme A) or with the presumably less myelotoxic sequence weekly cisplatin day 2, 9, 16, 30, 37, 44 (scheme B). In scheme C, CrEL 12 ml was administered prior to cisplatin in the sequence of Scheme A. 18 patients have received a total of 85 courses. In scheme A 4/10 patients, all treated with topotecan 0.45 mg m(-2) d(-1), experienced DLT: 1 patient had vomiting grade 4, 1 patient had grade 4 neutropenia >5 days, 1 patient had >2 weeks delay due to thrombocytopenia and 1 patient due to neutropenia. Both patients in scheme B (topotecan 0.45 mg m(-2) d(-1)) had DLT due to a delay > 2 weeks because of prolonged haematological toxicity. No DLT was observed in the first 3 patients in scheme C (topotecan 0.45 mg m(-2) d(-1)). However, 2 out of 3 patients treated at dose level topotecan 0.60 mg m(-2)d(-1) in scheme C experienced DLT due to >2 weeks delay because of persistent thrombocytopenia or neutropenia. We conclude that there is a modest clinical effect of CrEL on haematological toxicity for this cisplatin-based combination regimen, which seems to reduce these side effects but does not really enable an increase of the oral topotecan dose.

Measurement of pyruvate in blood by high-performance liquid chromatography with fluorescence detection.

Accurate measurement of pyruvate is particularly important in the detection of mitochondrial enzyme disorders. describe a simple high-performance liquid chromatography assay for pyruvate with derivatization and fluorescence detection. Between-run variation is below 4.5% and the method is linear between I and 1,200 micromol/L. The mean recovery is 101%. After sample pretreatment, the calculated pyruvate concentration proved to be stable for 15 days when samples were kept at -20 degrees C.

Phase I pharmacologic study of oral topotecan and intravenous cisplatin: sequence-dependent hematologic side effects.

PURPOSE: In in vitro studies, synergism and sequence-dependent effects were reported for the combination of topotecan and cisplatin. Recently, an oral formulation of topotecan became available. This phase I study was performed to assess the feasibility of the combination of oral topotecan and cisplatin, the pharmacokinetic interaction, and sequence-dependent effects. PATIENTS AND METHODS: Topotecan was administered orally (PO) daily for 5 days in escalating doses and cisplatin was given intravenously (IV) at a fixed dose of 75 mg/m(2) either before topotecan administration on day 1 (sequence CT) or after topotecan administration on day 5 (sequence TC) once every 3 weeks. Patients were treated in a randomized cross-over design. RESULTS: Forty-nine patients were entered onto the study; one patient was not eligible. Sequence CT induced significantly more severe myelosuppression than did sequence TC, and the maximum-tolerated dosage of topotecan in sequence CT was 1.25 mg/m(2)/d x 5. In sequence TC, the maximum-tolerated dosage of topotecan was 2.0 mg/m(2)/d x 5. Dose-limiting toxicity consisted of myelosuppression and diarrhea. Pharmacokinetics of topotecan and cisplatin were linear over the dose range studied; no sequence-dependent effects were observed. In addition, topotecan did not influence the protein binding of cisplatin or the platinum-DNA adduct formation in peripheral leukocytes in either sequence. CONCLUSION: The recommended dosages for phase II studies involving patients like the patients in our study are topotecan 1.25 mg/m(2)/d PO x 5 preceded by cisplatin 75 mg/m(2) IV day 1 once every 3 weeks, and topotecan 2.0 mg/m(2)/d PO followed by cisplatin 75 mg/m(2) IV day 5. No pharmacokinetic interaction could be discerned in our study. The antitumor efficacy of both schedules should be evaluated in a randomized phase II study.

Decrease of free thyroxine levels after controlled ovarian hyperstimulation.

Controlled ovarian hyperstimulation could lead to opposing effects on thyroid function. Therefore, in a prospective study of 65 women undergoing controlled ovarian hyperstimulation, thyroid hormones, T4-binding globulin, TPO antibodies, gonadotropins, estradiol, and PRL were measured before and after controlled ovarian hyperstimulation. After ovarian stimulation (mean +/- SE of mean): free T4 decreased, 14.4 +/- 0.2 vs. 12.9 +/- 0.2 pmol/L (P < 0.0001); thyroid-stimulating hormone increased, 2.3 +/- 0.3 vs. 3.0 +/- 0.4 mU/L (P < 0.0001); T4-binding globulin increased, 25.2 +/- 0.7 vs. 33.9 +/- 0.9 mg/L (P < 0.0001); total T4 increased, 98.1 +/- 2.3 vs. 114.6 +/- 2.5 nmol/L (P < 0.0001); total T3 increased, 2.0 +/- 0.04 vs. 2.3 +/- 0.07 nmol/L (P < 0.0001); TPO antibodies decreased, 370 +/- 233 U/mL vs. 355 +/- 224 U/mL (P < 0.0001); LH decreased, 8.1 +/- 1.1 vs. 0.4 +/-0.1 U/L (P < 0.0001); FSH did not change, 6.5 +/- 0.6 vs. 7.9 +/- 0.9 U/L (P = 0.08); human CG increased, <2 +/- 0.0 vs. 195 +/- 16 U/L (P < 0.0001); estradiol increased, 359.3 +/- 25.9 pmol/L vs. 3491.8 +/-298.3 pmol/L (P < 0.0001); and PRL increased, 0.23 +/- 0.02 vs. 0.95 +/- 0.06 U/L (P < 0.0001). Because low maternal free T4 and elevated maternal thyroid-stimulating hormone levels during early gestation have been reported to be associated with impaired psychomotor development in the offspring, our findings indicate the need for additional studies in the children of women who where exposed to high levels of estrogens around the time of conception.

Thyroid autoimmunity and abortion: a prospective study in women undergoing in vitro fertilization.

OBJECTIVE: To determine whether an association exists between the presence of thyroid peroxidase (TPO) antibodies before pregnancy and miscarriage in women without a history of habitual abortion. DESIGN: Prospective study and nested case-control study. SETTING: Inner-city teaching hospital. PATIENT(S): Four hundred eighty-nine women in an IVF program. INTERVENTION(S): In the prospective study, we measured levels of TPO antibodies and TSH. In the nested case-control study, we also measured levels of anticardiolipin antibodies. MAIN OUTCOME MEASURE: Miscarriage. RESULT(S): One hundred seventy-three women were observed, of whom 31% (54/173) became pregnant. Pregnancy occurred in 48% (12/25) of the antibody-positive women and in 28% (42/148) of the antibody-negative women. Among those who became pregnant, miscarriage occurred in 33% (4/12) of TPO antibody-positive women and in 19% (8/42) of TPO antibody-negative women. The TSH level was abnormal (<0.2 microIU/mL) in only one of the TPO antibody-positive women who miscarried. The presence of anticardiolipin antibodies was not associated with miscarriage. CONCLUSION(S): No association was found between the presence of TPO antibodies before pregnancy and miscarriage in women without a history of habitual abortion. The presence of TPO antibodies did not adversely affect a woman's chances of becoming pregnant.

The effect of herbal extracts in an experimental mouthrinse on established plaque and gingivitis.

The purpose of the present study was to establish in vitro the inhibiting effect of a herbal extract mixture on a selected number of micro-organisms and to test in vivo the effect of a mouthwash containing 6.3 mg/ml herbal extract mixture on plaque and gingivitis as compared to a minus active control mouthrinse. The herbal extract was a mixture of: Juniperus communis (juniper), Urtica dioca (nettle), Achillaea millefolium (yarrow); 1:1:1. In the study, in-vitro, the effect of pure herbal extract mixture on acid production of Streptococcus mutans was tested and the minimum inhibitory concentrations (MIC) of the following micro-organisms were tested: Streptococcus mutans, Streptococcus mitis, Actynomyces viscosus, Actynomyces naeslundii, Actinobacillus actinomycetemcomitans, Prevotella intermedia, Campylobacter rectus, Fusobacterium nucleatum, Veillonella parvula. The MIC-values for A. viscosus and P. gingivalis were 100 mg/ml. The MIC-values for A. naeslundii and A. actinomycetemcomitans were considerably lower (10 mg/ml). S. mitis was the most susceptible of the tested organisms to the extract with a MIC value of 1 mg/ml. S. mutans, C. rectus, V. parvula, and F. nucleatum were not influenced by the extracts. No inhibitory effect of the 6.3 mg/ml herbal extract mixture was observed on the acid production of S. mutans. For the study in-vivo, 45 volunteers were selected on the basis of having moderate gingival inflammation. As efficacy parameters the plaque index, modified gingival index and angulated bleeding index were assessed. The subjects were randomly divided among 3 experimental groups (2x test and 1 'minus active' control). The participants were requested to rinse with 10 ml of mouthwash twice a day for a period of three months. After 6 weeks and 3 months, the same clinical indices as at baseline were recorded. The results show no difference between the two test groups and the control group. In conclusion, the results of the present study have shown that the mixture of the 3 herbal extracts, Juniperus communis, Urtica dioca and Achillaea millefolium when used in a mouthrinse has no effect on plaque growth and gingival health.

Pharmacokinetics of the enantiomers of bupivacaine and mepivacaine after epidural administration of the racemates.

UNLABELLED: We investigated the pharmacokinetics of the enantiomers of bupivacaine and mepivacaine after epidural injection of the racemate of each drug into six surgical patients. After epidural administration of either bupivacaine/HCl (115 mg) or mepivacaine/HCl (460 mg), blood samples were collected for 24 h. Unbound fractions were determined by using ultrafiltration for bupivacaine and equilibrium dialysis for mepivacaine. Concentrations in plasma, ultrafiltrate, and dialysate were determined by using stereoselective high-performance liquid chromatography. Peak plasma concentrations of R(+)-bupivacaine (389 +/- 93 ng/mL) and R(-)-mepivacaine (1350 +/- 430 ng/mL) were smaller than those of S(-)-bupivacaine (449 +/- 109 ng/mL, P < 0.0001) and S(+)-mepivacaine (1740 +/- 490 ng/mL, P < 0.002), respectively. However, the unbound peak concentrations of R(+)-bupivacaine (20 +/- 11 ng/mL) were larger than those of S(-)-bupivacaine (15 +/- 9 ng/mL, P < 0.005); unbound peak concentrations of R(-)-mepivacaine (485 +/- 158 ng/mL) and S(+)-mepivacaine (460 +/- 139 ng/mL) did not differ. These observations reflect differences in the systemic disposition (distribution and elimination) of the enantiomers, because the systemic absorption was not enantioselective with either drug. This study supports the opinion that the use of single enantiomers, rather than racemates, is preferable, particularly for bupivacaine. IMPLICATIONS: Measurements of the plasma concentrations of the enantiomers of bupivacaine and mepivacaine after epidural administration of the racemates demonstrated that the systemic disposition, but not the systemic absorption, of these drugs is enantioselective and supports the opinion that the use of single enantiomers, rather than racemates, is preferable.

Evaluation of the long-term efficacy and safety of locally-applied minocycline in adult periodontitis patients.

The objectives of the present study were to establish in a long-term investigation the safety as well as the clinical and microbiological efficacy of scaling and rootplaning combined with local application of 2% minocycline hydrochloride-gel versus placebo-gel in patients with moderate to severe chronic adult periodontitis. This was an 18 months, randomized, double-blind, parallel, comparative study, in which 20 healthy patients with moderate to severe chronic periodontitis participated. At baseline, all patients received professional oral hygiene-instruction and supra- and subgingival scaling and root planing. The minocycline-gel was applied subgingivally baseline, 2 weeks, 1, 3, 6, 9 and 12 months. Microbiological evaluation was carried out using DMDx to identify the following bacteria: Porphyromonas gingivalis, Prevotella intermedia, Actinobacillus actinomycetemcomitans, Campylobacter rectus, Fusobacterium nucleatum and Treponema denticola. In addition standard microbiological techniques were used for the detection of P. gingivalis, P. intermedia, P. micros, A. actinomycetemcomitans, C. rectus, F. nucleatum, C. albicans and Enterobacteriaceae. Results showed a statistically significant improvement for all clinical parameters irrespective of the treatment modality. No differences were observed between test and control with regard to probing depth and attachment level. The DMDx data showed a significant reduction in both the numbers and the prevalence over the 15 months period, but no significant difference between groups. Culture data showed that at baseline two-third were positive for P. gingivalis and P. intermedia. Analysis over the 18 month period showed no significant difference between the two treatment modalities. C. albicans and Enterobacteriaceae were detected only in small proportions at each time interval in a limited number of patients. No adverse reactions were observed during the trial period. The present patient group responded favourably to scaling and rootplaning, but did not benefit from an effect of local of minocycline. Subgingival debridement in combination with oral hygiene instruction by itself has been shown to be effective. It remains to be studied whether local application of minocycline can be effective as an adjunct to mechanical therapy in sites that respond poorly to conventional treatment.

The effectiveness of an electronic toothbrush in the removal of established plaque and treatment of gingivitis.

The purpose of the present study was to evaluate the effect of an electronic toothbrush on established plaque and gingivitis in a 5-month clinical trial. This electronic toothbrush sends approximately a 0.15 mA current through the brushhead which presumedly enhances the efficacy of the brush in plaque elimination. For this study, 80 volunteers (non-dental University students) were selected on the basis of having moderate gingival inflammation. At baseline, plaque and bleeding upon probing were assessed on the vestibular, mesio-vestibular, distovestibular and lingual surfaces using the 'half-mouth' design. After 2-months and 5-months, these clinical indices were again recorded. At each assessment, the participants received a new brushhead. All baseline indices appeared to be well-balanced. A mean Silness and Löe plaque score of 1.70 and 1.64 were found in the control and test groups, respectively. Little change was observed from baseline to 5-months. The Quigley and Hein plaque index behaved comparably. The mean bleeding upon probing score at baseline was 1.43 and 1.39 for the control and test group, respectively. Little to no improvement was observed in the course of this study. No beneficial effect could be shown for this 'electronic' design of manual toothbrush.

Cystatins S and C in human whole saliva and in glandular salivas in periodontal health and disease.

Cystatins are inhibitors of cysteine proteinases and could play a protective and regulatory role under inflammatory conditions. Since total cystatin activity of whole saliva was increased in periodontal patients (Henskens et al., 1993), we wanted to investigate the types or origins of cystatins involved in this increase. Distinct types of cystatins were identified by isoelectric focusing and immunoblotting with specific antibodies against one of the salivary acidic isoforms, cystatin S. and the widely distributed basic cystatin C. Clarified human whole saliva (CHWS) of healthy subjects contained cystatin S, whereas cystatin C was barely detectable. In contrast, in CHWS of gingivitis and periodontitis patients, both cystatin C and S levels were higher. The origin of cystatin activity was investigated by collecting submandibular (SM), sublingual (SL), and parotid (PAR) saliva from seven subjects with mild gingivitis. Total cystatin activity was about five times higher in SM saliva than in PAR saliva. In SM and SL saliva, both cystatins S and C were demonstrated. In contrast, in PAR samples, solely cystatin C was detectable. The introduction of experimental gingivitis in one periodontally healthy subject resulted in the appearance of a cystatin C band in PAR saliva and in an increase of cystatins S and C in SM saliva. We conclude that the previously observed increase of cystatin activity in whole saliva in inflammatory periodontal disease is, at least in part, due to an increased glandular output of both the isoform cystatin S (pI 4.7) and the basic cystatin C (pI 9.0).

Formation and elimination of sulphamethoxazole hydroxylamine after oral administration of sulphamethoxazole.

The formation and elimination of sulphamethoxazole hydroxylamine in relation to the pharmacokinetics of the parent compound and its N4-acetyl metabolite were investigated in six healthy subjects after a single oral dose of 800 mg sulphamethoxazole. The apparent half-lives of sulphamethoxazole and its metabolites were approximately 10 h, indicative of formation rate-limited metabolism. The mean residence time of the hydroxylamine metabolite was 5.5 +/- 1.5 h. The renal clearance of sulphamethoxazole hydroxylamine was 4.39 +/- 0.91 l h-1. The urinary recovery of sulphamethoxazole accounted for 16.5 +/- 5.5% of the dose, N4-acetyl-sulphamethoxazole for 46.2 +/- 6.6% and the hydroxylamine metabolite for 2.4 +/- 0.8%. The remaining 35% of the dose was unaccounted for. Acetylator phenotype was determined using sulphadimidine. The renal excretion of sulphamethoxazole hydroxylamine was 1.9 +/- 0.9% in slow acetylators (n = 3) and 2.8 +/- 0.3% in fast acetylators (n = 3); for N4-acetyl-sulphamethoxazole the values were 48 +/- 6% and 44 +/- 8%, respectively. Sulphamethoxazole is metabolized, although to a limited extent, to a hydroxylamine metabolite. This metabolite may be important for the pathogenesis of adverse reactions.

The long-term effect of an oscillating/rotating electric toothbrush on gingivitis. An 8-month clinical study.

The purpose of this study was to evaluate the safety and efficacy of the Braun Plak Control for the removal of supragingival plaque and improving gingival health in a long-term clinical trial, and to compare it to regular manual toothbrush. Assessed were plaque accumulation, amount of gingival inflammation, gingival bleeding on probing, and calculus. In total, 77 young individuals were selected on the basis of having 'moderate gingivitis'. They were monitored over 8 months and divided among 2 groups; a control group that used a manual toothbrush and a test group that used the Braun Plak Control. The clinical assessments were repeated after 1, 2, 5, and 8 months. At baseline, subjects were handed their assigned toothbrushes together with written oral hygiene instructions. They were instructed to brush for at least 2 min. 1 month after baseline examinations, all subjects received a professional prophylaxis and oral hygiene instruction from an experienced dental hygienist. Plaque removal was reinforced at the 2-and 5-month examination. In conclusion, results indicate that the Braun Plak Control is a safe and efficient home care device. At the end of this trial, this electric toothbrush proved to be more effective than a regular manual toothbrush.

Preparing your organization for technological change.

It is not always easy to know exactly why a particular group resists change. However, experience shows that an intelligent application of this basic five-step change process--coupled with a sound technological implementation plan--leads to more rapid and more productive introductions of technology into organizations. The process can be expensive in terms of time and energy but nowhere near the cost of an expensive technical system that never gains user acceptance.

Effect of selenite on the toxicity of cis-DDP in mice: estimation of trace elements.

A significant accumulation of copper, zinc, iron, rubidium and bromine, in addition to platinum, was observed in kidneys and liver of mice treated with cis-DDP. This observation suggests that the toxicity of cis-DDP is due to the overall accumulation of trace elements and not only to the expected high platinum levels. Sodium selenite administration prior to cis-DDP results in fast clearance of all these trace elements from kidneys and liver; control levels are reached within 6 days. This may explain the important reduction in cis-DDP toxicity following selenite administration. An X-ray fluorescence (XRF) facility was used for the evaluation of the trace elements.