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.

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.

Temporal relation of changes in regional coronary flow and myocardial lactate and nucleoside metabolism during pacing-induced ischemia.

The temporal relation between myocardial lactate and hypoxanthine metabolism and regional changes in krypton-81m perfusion during pacing-induced ischemia was studied in 17 patients with coronary artery disease (CAD). During incremental atrial pacing, lactate production and hypoxanthine release occurred early and simultaneously, accompanied by ST-segment changes, but before angina and only few minutes after a significant (17%) reduction in krypton-81m perfusion in areas with more than 90% luminal diameter reduction. During maximal pacing heart rates, krypton-81m distribution decreased to 68 +/- 7% of control in areas with more than 90% diameter reduction and to 80 +/- 4% in 70 to 90% reduction (both p less than 0.05 vs control). Maximal lactate production occurred 15 seconds after pacing (extraction -15 +/- 7% vs 16 +/- 2% during control, p less than 0.05) and peak hypoxanthine release 1 minute after pacing (delta arteriovenous -2.64 +/- 0.8 microM vs 0.08 +/- 0.21 microM during control, p less than 0.05). Krypton-81m perfusion decreased in 20 of the 21 CAD areas. Angina, ST-segment changes, hemodynamic alterations and lactate production occurred in 15, 14, 9 and 15 patients, respectively. In contrast, hypoxanthine release was found in all cases. After pacing, lactate production and all general indexes of ischemia persisted for only 2 to 3 minutes. In contrast, krypton-81m perfusion was still significantly reduced 5 minutes after pacing and was only accompanied by hypoxanthine release (delta arteriovenous -1.41 +/- 0.6 microM, p less than 0.05 vs control).(ABSTRACT TRUNCATED AT 250 WORDS)

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.

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).

Determination of bromine in blood serum by 125I excited x-ray fluorescence.

A radioisotope excited XRF method based on the use of five 10-mCi 125I point sources arranged in a newly designed configuration is applied to the bromine determination in very small blood serum samples (less than or equal to 50 microL). The possibility of determination of 0.05 micrograms Br in blood serum during 100 s counting time with a standard deviation of +/- 10% is described. The simplicity and low cost of the present method are major advantages over the other nondestructive methods and render it suitable for medical laboratories.

A rapid quantitative determination of triglycerides by means of thin-layer chromatography.

A quantitative chromatographic method for determining triglycerides in serum is described. The serum is applied directly to a thin-layer plate and after partition with one developing solvent the fractions are charred with 10% sulphuric acid. Charring takes 20 minutes. The concentration of triglycerides is measured against a triolein standard, which in itself is corrected for having too many double bonds by scanning the charring of the triglyceride fraction with a "flying spot"-scanner or a slitscanner. The results are compared with other methods of determination.

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.

The distribution of selenium in human blood samples of Israeli population--comparison between normal and breast cancer cases.

A preliminary study was carried out in order to compare the selenium concentration in breast cancer patients and healthy subjects (controls) in Israel. Blood serum samples were obtained from 32 breast cancer patients and 36 controls and were analyzed for selenium by the XRF method. A weighted mean of 0.076 +/- 0.014 ppm Se in the blood serum of breast cancer patients, as compared to 0.119 +/- 0.023 ppm Se for controls, was obtained. These results indicate that the concentration of selenium in breast cancer patients is significantly lower than in controls. The relationship between selenium concentration and malignancy stage shows an inverse dependence, i.e., the concentration decreases with stage number.

Rapid and simple determination of selenium and other trace elements in very small blood samples by XRF.

Selenium and other trace elements (Cu, Zn, Br, and Rb) were determined in very small (0.75 microL) human serum and mice whole blood samples, by an XRF method. Accurate results of elemental concentration were obtained without the need of exact volume measurement, because of the backscatter correction used. The XRF method is highly sensitive (M.D.L. = 0.06, 0.13, 0.09, 0.07, and 0.05 ppm for Se, Cu, Zn, Br, and Rb, respectively), rapid (counting time--100 s/sample), easy to perform and therefore suitable for routine trace element analyses. The results obtained are in good agreement with the values reported in the literature.

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.

Reversibility of the ATP:glutamine synthetase adenylyltransferase reaction.

The reversibility of adenylylation of glutamine synthetase from E. coli by adenylyltransferase was demonstrated. Several positive effectors (Gln, 2-hydroxyethyl-S-cysteine, Trp and Met) stimulate the back reaction in the same manner as the forward reaction. The apparent Michaelis constant for PP(i) is 2.2 mM at pH 7.35. The pH optimum of the back reaction is 6.5-7 while the pH optimum of the forward reaction is 7.6. The apparent equilibrium constant in the presence of 10 mM Mg(2+) at pH 7.36 is 8.5 in favor of adenylylated glutamine synthetase and PP(i). The equilibrium constant is strongly dependent from pH and from Mg(2+) concentration. There is a difference of about 0.5 to 1 kcal/mole free energy between the adenylyl-O-tyrosine bond and the pyrophosphate bond of adenosine triphosphate (ATP). It follows from these considerations that the adenylyl-O-tyrosine bond is an "energy-rich phosphate bond."