Pharmacological characterization of muscarinic receptors implicated in rabbit detrusor muscle contraction and activation of inositol phospholipid hydrolysis in rabbit detrusor and parotid gland

In the present study, we evaluated the pharmacological characteristics of the functional muscarinic receptors implicated in rabbit detrusor contraction and coupled to inositol phospholipid turnover in rabbit detrusor and parotid gland. The selectivity of several muscarinic antagonists for detrusor vs. salivary gland muscarinic receptors was also examined. The affinities for the muscarinic m1-, m2- and m3-receptor subtypes were determined using membranes from human cloned receptors expressed in CHO-K1 cells using [3H]-N-methyl scopolamine as a radioligand. Anti-muscarinic activity was determined in isolated rabbit detrusor by measuring the displacement of the contractile response to carbachol, and in rabbit detrusor and rabbit parotid by measuring the displacement of inositol phospholipid hydrolysis (total inositol phosphate accumulation) to carbachol. A significant correlation was found between the potencies to antagonize carbachol-induced rabbit detrusor contraction (pK(B)) and the affinities (pKi) for the m3-receptor subtype (r = 0.93, P = 5 x 10(-6)). Lower, but significant, correlations [0.88 (P = 6.3 x 10(-5)), 0.72 (P = 4.6 x 10(-3))] were obtained with m1- or m2-receptor subtypes, respectively. Each muscarinic antagonist tested displayed similar potency to antagonize carbachol-stimulated inositol phospholipid hydrolysis in rabbit detrusor and parotid (r = 0.96, P = 8 x 10(-3)). A significant correlation was found between the potencies to antagonize carbachol-stimulated inositol phospholipid hydrolysis (pK(B)), determined in rabbit detrusor and rabbit parotid, and the affinities (pK(i)) for the m3-receptor subtype [r = 0.96 (P = 0.01), 0.99 (P = 5 x 10(-5)), respectively] and for the m1-receptor subtype [r = 0.98 (P = 3.5 x 10(-3)), 0.94 (P = 0.02), respectively] but not for the m2-receptor subtype [r = 0.33, 0.57, ns, respectively]. In each in vitro assay, methoctramine (preferential M2 selective antagonist) and pirenzepine (preferential M1 selective antagonist) were slightly potent. We suggest that the muscarinic receptor implicated in the response to carbachol in rabbit detrusor and parotid gland corresponds to the M3-subtype. None of the muscarinic antagonists studied in rabbit tissues displayed preferential affinity for the detrusor.     

Investigation of Nonimmune Hydrops Fetalis: Multidisciplinary Studies Are Necessary for Diagnosis—Review of 94 Cases

This review of 94 cases of nonimmune hydrops fetalis (NIHF) over a 10-year period was undertaken to evaluate the frequency of this pathology among fetal and infant deaths and to determine the most common likely etiologies in a northeastern region of France. NIHF represented 6% of the fetal deaths examined in our laboratory. The combination of findings from morphologic examination of the placenta and fetus with the results of microbiological and cytogenetic investigations (conventional cytogenetic study, fluorescent in situ hybridization [FISH], or DNA ploidy image analysis) led to an etiologic diagnosis for NIHF in two-thirds of the cases and suggested a diagnosis in an additional 23% of cases. The most common causes of NIHF were chromosome abnormalities (33%), infections (16%), and cardiac pathology (13.8%). The detection of a cause for NIHF is important for genetic counseling and management of subsequent pregnancies. Our experience suggests that a diagnosis is possible in a large majority of NIHF when obstetricians and pathologists carefully coordinate the management of prenatal and postnatal investigations and when new techniques, such as molecular biology and DNA quantification, are used. Received January 12, 1998; accepted October 14, 1998.     

Acute lower gastrointestinal bleeding in Crohn's disease: characteristics of a unique series of 34 patients

OBJECTIVE: Acute lower gastrointestinal bleeding is a rare complication of Crohn's disease, which represents a diagnostic and therapeutic challenge. The aim of this study was to define epidemiological characteristics and therapeutic options of hemorrhagic forms of Crohn's disease. METHODS: Thirty-four cases of hemorrhagic forms of Crohn's disease were studied retrospectively. Acute lower gastrointestinal hemorrhage was defined as acute rectal bleeding originating in diseased bowel and requiring a transfusion of at least 2 units of red blood cells within 24 h. Upper gastrointestinal tract hemorrhage or anal lesions and postoperative bleeding were excluded. RESULTS: Mean age at time of hemorrhage was 34.2 +/- 14 yr. Mean duration of disease before the hemorrhage was 5.6 +/- 6 yr. The hemorrhage occurred during a flare up of the disease in 35% of cases. The hemorrhage revealed Crohn's disease in 23.5% of cases. The hemorrhage was more frequent in colonic disease (85%) than in isolated small bowel disease (15%) (p < 0.0001). The origin of bleeding was identified in 65% of cases, by colonoscopy (60%), by angiography (3 patients), or at surgery (1 patient). The bleeding lesion was an ulcer in 95% of cases, most often in the left colon. The treatment was surgical in 20.5% (colectomy in 36%), endoscopical (7 patients, including 5 successes), or medical. Hemorrhage recurred in 12 patients (35%) within a mean time of 3 yr (4 days-8 yr), requiring surgery in 3 cases. No death was observed. CONCLUSIONS: This study performed in a series characterized by a nonsurgical recruitment, the largest to date, shows that hemorrhagic forms of Crohn's disease may reveal disease in 23.5%, occurs in quiescent Crohn's disease in two-thirds of cases. Given the potential efficacy of endoscopical or medical treatment, as well as the absence of mortality, a conservative approach may be suggested as first-line therapy in the majority of patients.     

Design and structure-activity relationship analysis of ligands of gamma-hydroxybutyric acid receptors

With the use of [³H]gamma-hydroxybutyric acid, binding experiments allowed the screening of new compounds as ligands of gamma-hydroxybutyric acid receptors. Starting from the acid-alcohol gamma-hydroxybutyric acid structure, structure–activity relation analysis and lead optimization highlighted gamma-hydroxybutyric acid derivatives with significantly increased affinities, when compared with the affinity of gamma-hydroxybutyric acid. Further pharmacological studies with the use of gamma-hydroxybutyric acid derivatives allowed the characterization of the first competitive antagonist acting at gamma-hydroxybutyric acid receptors (NCS 382).     

Adolescent physical performance and adult physical activity in Flemish males

Limited information is available about the associations between adolescent fitness levels and adult physical activity. In the present study, these associations are investigated using different indicators of physical activity. It is hypothesized that both health‐ and performance‐related fitness characteristics, observed during the adolescent period, contribute equally to the explained variance in adult physical activity levels. Subjects were 109 Flemish males followed over a period of 27 years from 13 to 40 years of age in the Leuven Longitudinal Study on Lifestyle Fitness and Health. Performance and health‐related fitness characteristics were observed during the growth period and at 40 years of age. The Work Index, Leisure Time Index, and Sport Index of the Baecke questionnaire were used as indicators of physical activity together with triaxial accelerometry. Multiple regression and discriminant analyses contrasting extreme quintiles of activity groupings were used to analyse the associations. Only the Baecke Sport Index showed consistent significant associations (R² = 0.03 to R² = 0.23) with adolescent fitness levels observed at 13, 15, and 18 years. When upper and lower quintiles were contrasted, fitness characteristics observed at the three age levels during adolescence were significantly different for each of the three indices of the Baecke questionnaire at 40 years of age. Lowest associations (R² = 0.09 to R² = 0.17) were found for the Work Index, followed by the Leisure Time Index (R² = 0.12 to R² = 0.28) and Sport Index (R² = 0.25 to R² = 0.43). Highest associations were evident for the 18‐ to 40‐year interval. Performance‐ and health‐related fitness characteristics explain equally well the variance in physical activity indicators. Am. J. Hum. Biol. 13:173–179, 2001. © 2001 Wiley‐Liss, Inc.     

Clinical variability and genetic homogeneity of the camptodactyly‐arthropathy‐coxa vara‐pericarditis syndrome

The camptodactyly‐arthropathy‐coxa vara‐pericarditis syndrome (CACP) is an autosomal recessive condition characterized by the association of congenital or early onset camptodactyly and noninflammatory arthropathy with synovial hyperplasia. Progressive coxa vara deformity and/or noninflammatory pericardial or pleural effusions have been observed in some patients. Recently, the disease gene has been assigned to human chromosome region 1q25‐q31, and truncating mutations have been identified in the megakaryocyte stimulating factor gene. Studying 12 patients from 8 unrelated families, we emphasized hip and spine involvement, particularly in the course of the disease as shown in a 58‐year‐old patient. Despite clinical variability, linkage studies support genetic homogeneity of the disease. Am. J. Med. Genet. 95:233–236, 2000.     

Relation of smoking to breast cancer by estrogen receptor status

It has been suggested that smoking is associated with estrogen-negative breast cancer but not with estrogen-positive breast cancer. A population-based case-control study was conducted in Geneva, Switzerland, to determine the relation of passive and active smoking to breast cancer when the referent unexposed category consisted of women unexposed to active and passive smoke. The 242 patients with breast cancer (cases), in whom estrogen receptor (ER) status was determined on biopsy material, were compared with 1,059 women free of breast cancer (controls). Lifetime history of active and passive smoking was recorded year by year, between age 10 and the date of interview. Prevalence rates of ER⁺ tumors were 74.7% in pre-menopausal women and 74.2% in post-menopausal women. Post-menopausal active smokers had a lower prevalence of ER⁺ tumors (70.0%, p = ns). Among pre-menopausal women, the age-adjusted ORs of breast cancer with having smoked an average of ≥20 cigarettes per day (cpd) during lifetime were 2.7 for ER⁻ tumors and 2.6 for ER⁺ tumors. Among post-menopausal women, corresponding ORs were 5.7 for ER⁻ tumors and 2.4 for ER⁺ tumors. Smoking was related to both ER⁻ and ER⁺ breast cancer in pre- and post-menopausal women, but the strength of the association appeared to be greater for ER⁻ tumors among post-menopausal women. Int. J. Cancer 75:339–342, 1998. © 1998 Wiley-Liss, Inc.     

Resistance to high concentrations of methotrexate and 5-fluorouracil of differentiated HT-29 colon-cancer cells is restricted to cells of enterocytic phenotype

Adaptation of HT-29 cells to increasing concentrations of methotrexate (MTX) results in the selection of differentiated populations which show sequential dose-dependent changes of their differentiated phenotype with, at the highest concentrations (0.1 and 1 mM), a shift of differentiation from a mucus-secreting to an enterocytic phenotype coinciding with an amplification of the DHFR gene. We show here that DHFR gene amplification itself does not play a role in the shift of differentiation. An alternative explanation is the presence, within the mucus-secreting population, of an undetectable minor population of cells committed to enterocytic differentiation and able to develop resistance to higher concentrations of MTX. This was confirmed by cloning the population of cells resistant to 10 μM MTX. Out of 19 isolated clones, 17 were found to be mucus-secreting and 2 enterocytic. We tested 9 of these clones for their ability to develop resistance to 0.1 mM MTX: only 1 of enterocytic phenotype, was found to develop resistance to this higher concentration and to amplify the DHFR gene. The ability of enterocytic cells to develop resistance to elevated MTX concentration through amplification of the DHFR gene was demonstrated in another enterocytic HT-29 population selected by glucose deprivation. Enterocytic cells resistant to 10 μM MTX were also found, unlike mucus-secreting cells, to be readily adaptable to 5-fluorouracil, this occurring without amplification of the thymidylate synthase gene. Together these results highlight a previously uncharacterized relationship between commitment to enterocytic differentiation of colon-cancer cells and their ability to develop resistance to MTX and 5-fluorouracil. Int. J. Cancer 76:383–392, 1998.© 1998 Wiley-Liss, Inc.