3,3-Bis-(4-hydroxyphenyl)-7-methyl-2-indolinone (BHMI), the active metabolite of the laxative sulisatin

The disodium salt of the sulphuric diester of 3,3-bis-(4-hydroxyphenyl)-7-methyl-2-indolinone (sodium sulisatin, Laxitex), a synthetic laxative with two phenolic groups esterified with sulfate, has been studied in order to find out if its laxative properties may be attributed to the unaltered compound or to its diphenolic derivative BHMI. We first studied the effect of homogenates of the gastrointestinal tract of rats and of rat cecal content of the hydrolysis of sulfate ester bonds of sulisatin. Results show that sulisatin can be hydrolyzed by cecal content while homogenates of stomach, small intestine and large intestine have no hydrolytic effect. Sulisatin is also a substrate of arylsulfate sulphohydrolase obtained from the snail Helix pomatia. The unaltered drug has no effect on the intestinal motility since it does not change the intestinal transit speed in rats pretreated with neomycin sulfate. Sulisatin (1.5, 3 and 6 mg) is unable to inhibit water absorption in rat colon while small amounts of BHMI (15 and 30 micrograms) may inhibit it significantly. It is concluded that sulisatin passes unaltered through the small intestine and is hydrolyzed in the large intestine by the intestinal microflora to its diphenolic derivative BHMI, which is responsible for the laxative activity of the drug.     

Amino-terminal pro-brain natriuretic peptide predicts ventricular arrhythmogenesis in patients with ischemic cardiomyopathy and implantable cardioverter-defibrillators

STUDY OBJECTIVES: Even in high-risk population groups, not all patients have the same risk of sudden cardiac death (SCD). Given the emerging data about the amino-terminal fragment of the brain natriuretic peptide prohormone (NT-proBNP) value in heart failure, we planned to evaluate the importance of NT-proBNP levels in predicting the occurrence of malignant arrhythmias in patients with ischemic cardiomyopathy and implantable cardioverter-defibrillators (ICDs). DESIGN: Prospective study. SETTING: Tertiary referral center. PATIENTS: Thirty five ambulatory patients with previous myocardial infarction, left ventricular ejection fraction < 35%, and ICDs for primary prevention of SCD according to Multicenter Automatic Defibrillator Implantation Trial I criteria. INTERVENTIONS: Venous blood samples for plasma NT-proBNP measurement were obtained after 30 min of supine rest from all patients at the beginning of the study. Patients were evaluated every 2 months, or sooner in cases of device discharges, during a 1-year follow-up period. Data concerning arrhythmias and device therapy were stored at the time of device interrogation on each follow-up visit. MEASUREMENTS AND RESULTS: During 1-year follow-up, 11 of 35 patients (31.4%) received 18 antiarrhythmic device therapies for ventricular tachyarrhythmia (VT). Patients who experienced such arrhythmias had NT-proBNP levels of 997.27 +/- 335.14 pmol/L (mean +/- SD), whereas those without VT had NT-proBNP levels of 654.87 +/- 237.87 pmol/L (p = 0.001). An NT-proBNP cutoff value of 880 pmol/L had a sensitivity of 73%, a specificity of 88%, a positive predictive value of 80%, and a negative predictive value of 88% for the prediction of occurrence-sustained VT events. CONCLUSION: To achieve the maximum benefit by ICD therapy, more precise risk stratification is required, even in high-risk, post-myocardial infarction patients. Plasma NT-proBNP levels comprise a promising method that could help in the better identification of a patient group with an even higher risk of sudden death.     

Proceedings of a consensus conference: the screening of blood donors for variant CJD

Since the identification, in 1996, of the first case of variant Creutzfeldt-Jakob disease (vCJD) in humans various approaches have been implemented and/or proposed to prevent this disease from being transfusion transmitted. In addition, a variety of possible laboratory-based approaches have been developed and will continue to be developed for the vCJD screening of blood donors. Various issues related to the implementation of such vCJD testing is likely to assume greater importance as diagnostic tests for vCJD becomes available for the potential screening of blood donors. The purpose of this Consensus Conference was to bring together international experts in an effort to determine which principles should guide the introduction of such testing. These experts provided the scientific and biological background of bovine spongiform encephalopathy (BSE) and vCJD, an understanding of their current epidemiology, as well as the ethical and legal issues that would impact on the implementation of a screening test for preventing the transfusion transmission of vCJD. This contentious issue is of potential considerable importance to transfusion medicine personnel worldwide, as well as to future recipients of allogeneic blood components.     

Development of an immunofluorometric assay and quantification of human kallikrein 7 in tissue extracts and biological fluids

BACKGROUND: Human kallikrein 7 (hK7), also known as human stratum corneum chymotryptic enzyme, is a chymotrypsin-like serine protease first identified in human skin extracts and predicted to be a secreted protease. The aim of this study was to develop a sensitive and specific immunoassay for hK7 and to examine the distribution of hK7 in tissue extracts and biological fluids. METHODS: Recombinant hK7 was produced in human embryonic kidney cells (HEK293T) and purified by a three-step column chromatographic procedure. The purified hK7 was injected into mice for antibody generation. A sandwich-type immunoassay was developed with the anti-hK7 monoclonal antibodies. RESULTS: The assay had imprecision (CV) <10% through the dynamic range of 0.2-20 microg/L and had no detectable cross-reactivity from other members in the human kallikrein gene family. Highest concentrations were found in skin, esophagus, and kidney. hK7 was also found in amniotic fluid, ascites from ovarian cancer patients, breast milk, cerebrospinal fluid, saliva, seminal plasma, serum, sweat, synovial fluid, and urine. CONCLUSIONS: This study describes the first ELISA-type immunoassay for hK7 protein quantification. hK7 is found many human tissues and in various biological fluids.     

Hair analysis differentiates chronic from acute carbamazepine intoxication

This is a report of a 12-year-old epileptic child undergoing chronic treatment with carbamazepine who was found comatose. He was considered to have acute severe drug toxicity. Measurement of carbamazepine concentration in the patient's hair segments together with the carbamazepine blood levels were both important in determining the chronic nature of the patient's intoxication.     

MIB1 as a possible predictor of recurrence in low-grade endometrial stromal sarcoma of the uterus

OBJECTIVE: Immunohistochemical analysis of MIB1, p53, estrogen, and progesterone receptors can provide prognostic information in endometrial adenocarcinoma. Since predictors of recurrence for low-grade endometrial stromal sarcoma (LESS) are still unknown, a battery of immunostains was performed to find markers, which might be useful to predict prognosis. METHODS: Eleven patients with an average age of 43.8 years (range 27-76) were identified with stage I LESS. Immunostains, including MIB1, p53, ER, and PR, were evaluated by two pathologists, independently. RESULTS: All tumors were positive for ER and PR; 1/11 was positive for p53; MIB1 ranged from 0 to 20% positive tumor nuclei. Mitotic counts ranged from 0 to 7/10 hpf. Two patients developed recurrences. One had a pelvic recurrence 7 years after diagnosis. This tumor had a mitotic count of 1/10 hpf, MIB1 expression in 10% of nuclei, and focal p53 expression. A second patient developed pulmonary metastases 10.8 years after diagnosis; the tumor showed a mitotic count of 7/10 hpf and MIB1 expression in 20% of nuclei, but was negative for p53. There was a significant difference in MIB1 reactivity scores between patients who did or did not develop recurrence (P = 0.0303). A marginally significant association was detected between MIB1 (P = 0.0896) or p53 (P = 0.0833) positivity and length of recurrence-free survival. CONCLUSION: Although MIB1 and p53 appear to be useful prognostic markers, a larger study would be necessary to confirm their validity.     

Flavonoids can block PSA production by breast and prostate cancer cell lines

BACKGROUND: Prostatic carcinoma is the most commonly diagnosed cancer and the second leading cause of cancer death of North American men. Combined androgen blockade (CAB) is one treatment option for prostate cancer, using estrogen agonists, luteinizing hormone-releasing hormone (LHRH) agonists and non-steroidal anti-androgens such as nilutamide and cyproterone acetate. Since many of these drugs have serious side effects, many patients are searching for "natural" alternatives or complements to traditional therapy. These include phytoestrogens found in soy and other plant foods. Such compounds have only started to be evaluated for potential androgen-blocking activity. Inhibition of production of androgen-regulated proteins, including prostate-specific antigen (PSA), is one indicator of androgen blocking. METHODS: The ability of 72 flavonoids and related compounds to inhibit PSA production in a breast cancer cell line, BT-474, and a prostate cancer cell line (PC-3), transfected with the human androgen receptor cDNA, PC-3(AR)(2) was examined. RESULTS: Twenty-two of the 72 flavonoids tested were found to significantly block PSA production by the BT-474 cell line at the highest tested concentration (10(-5) mol/l), with 17 of these compounds inhibiting production of PSA in the PC-3(AR)(2) cells as well. CONCLUSIONS: That several flavonoids may significantly block production of this androgen-regulated protein. It will be worthwhile to examine these compounds as possible candidates for prostate cancer prevention or management.