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Biomedical subjects

A Lambert

Publications and source records attributed to A Lambert.

At least 55 records · Page 3Linked to original sources
PubMed2000

Improved FSH sensitisation and aromatase assay in human granulosa-lutein cells.

Granulosa-lutein (GL) cells from follicular aspirates from women undergoing in-vitro fertilization (IVF) treatment are usually refractory to follicle stimulating hormone (FSH) regarding the induction and/or maintenance of aromatase activity which converts androgens (e.g. testosterone) to oestrogens. The normal method of assaying FSH-stimulated aromatase activity in GL cell cultures is to add exogenous testosterone throughout the cell culture period and measure the secreted oestradiol. Thus under the conditions usually employed for studying FSH-stimulated oestradiol secretion in GL cells, the 'total' FSH effect is dependent both on the decay of the aromatase concentration in culture relative to its induction/maintenance by FSH and on changes in its activity in the face of a declining substrate concentration as the exogenous testosterone is converted over several days to oestradiol. We have therefore used a technique for challenging the cells with testosterone (10 micromol/l) for just 2 h at the end of the normal longer-term culture period such that its concentration was essentially unchanged, thus ensuring that there was no depletion of the aromatase substrate and that the FSH stimulation phase could be performed independently of exogenous testosterone. Consequently, GL cells were incubated for 0, 24 and 48 h prior to stimulation with FSH (100 IU/l) for 24 h after which they were washed and challenged with testosterone for 2 h and the secreted oestradiol was assayed. Freshly isolated GL cells from women undergoing IVF were refractory to FSH but after preincubation were responsive such that there was a 3-14-fold increase over basal activity depending on the cell preparation. In conclusion, we have developed a simple 48 h procedure for sensitizing GL cells to FSH and established the conditions for optimizing the assay of aromatase activity independently from the effect of FSH on its induction.

Aromatase↗
PubMed2000

Characterization of a Snorhizobium meliloti ATP-binding cassette histidine transporter also involved in betaine and proline uptake.

The symbiotic soil bacterium Sinorhizobium meliloti uses the compatible solutes glycine betaine and proline betaine for both protection against osmotic stress and, at low osmolarities, as an energy source. A PCR strategy based on conserved domains in components of the glycine betaine uptake systems from Escherichia coli (ProU) and Bacillus subtilis (OpuA and OpuC) allowed us to identify a highly homologous ATP-binding cassette (ABC) binding protein-dependent transporter in S. meliloti. This system was encoded by three genes (hutXWV) of an operon which also contained a fourth gene (hutH2) encoding a putative histidase, which is an enzyme involved in the first step of histidine catabolism. Site-directed mutagenesis of the gene encoding the periplasmic binding protein (hutX) and of the gene encoding the cytoplasmic ATPase (hutV) was done to study the substrate specificity of this transporter and its contribution in betaine uptake. These mutants showed a 50% reduction in high-affinity uptake of histidine, proline, and proline betaine and about a 30% reduction in low-affinity glycine betaine transport. When histidine was used as a nitrogen source, a 30% inhibition of growth was observed in hut mutants (hutX and hutH2). Expression analysis of the hut operon determined using a hutX-lacZ fusion revealed induction by histidine, but not by salt stress, suggesting this uptake system has a catabolic role rather than being involved in osmoprotection. To our knowledge, Hut is the first characterized histidine ABC transporter also involved in proline and betaine uptake.

ATP-Binding Cassette Transporters↗
PubMed2000

Increased expression of growth hormone and prolactin receptors in hepatocellular carcinomas.

The liver is an essential target tissue for growth hormone (GH) and prolactin (PRL). The aim of this study was to determine the in situ expression of growth hormone receptor (GHR) and prolactin receptor (PRLR) in hepatocellular carcinomas and to compare the results with normal liver. For this purpose, in situ hybridization (ISH) and immunohistochemical techniques were performed and several tests were conducted to validate the results. By radioactive ISH, all the hepatocellular carcinomas studied showed labeling for GHR and PRLR mRNAs. Relative expression levels, determined by computer-assisted microdensity, were higher in hepatocellular carcinomas than in normal liver. Immunohistochemistry led us to confirm the constant expression of both receptor proteins in hepatocellular carcinomas and normal liver and to demonstrate their localization not only in the cytoplasm but also in the nucleus. These results confirm that the liver is a major GH and PRL target tissue and suggest that in hepatocellular carcinomas the proliferative effects of these hormones may be increased by a higher expression of their receptors.

Adult↗
PubMed2000

Host death: a necessary condition for the transmission of Aphalloides coelomicola Dollfus, Chabaud, and Golvan, 1957 (Digenea, Cryptogonimidae)?

Eggs of the trematode Aphalloides coelomicola were observed free in the abdominal cavity of Pomatoschistus microps from March 1993 to October 1995 (Pisces, Gobiidae) in the Vaccares lagoon (delta du Rhone, France). The eggs were contained in a fine membrane of parasitic origin as no immunological reaction of the fish was noted. The alternatively high prevalence of adults and low prevalence of free eggs, and vice versa, suggest that P. microps is both the second intermediate and final hosts of the trematode Aphalloides coelomicola and that the mechanism of transfer to the first intermediate host is through death of the definitive host.

Animals↗
PubMed1999

Influence of the trematode Aphalloïdes caelomicola Dollfus, Chabaud & Golvan, 1957 on the fecundity and survival of Pomatoschistus microps (Krøyer, 1838) (Teleostei: Gobiidae).

Monthly data collected during 1993 in the Vaccarès lagoon (Rhône Delta, France) were used to investigate the influence of the trematode Aphalloïdes coelomicola on the survival of the common goby Pomatoschistus microps, which acts both as second intermediate and definitive host. Prevalence of the parasite did not increase with fish size in either sex. Mean abundance and variance to mean abundance ratio gave evidence that the trematode could have an impact on host mortality. Gonad weight of parasitized females was reduced and the parasite seemed to provoke a gonadal regression above a certain parasitic biomass. Fecundity and egg diameters did not differ between parasitized and unparasitized females, we thus hypothesize that the energy content of eggs (composition in lipids and proteins) was probably modified as an adaptation to energy losses. Prevalence of infection in males compared to females and influence of the parasite on host survival and reproductive effort are discussed.

Analysis of Variance↗
PubMed1999

Seven new species of Quadriacanthus Paperna, 1961 (Monogenea) from the gills of Heterobranchus longifilis and H. isopterus from the Ivory Coast, west Africa.

The gills of two species of Siluriformes, Heterobranchus longifilis and H. isopterus collected from the Rivers Agnéby and Bia in the Ivory Coast (West Africa) revealed the presence of seven new species of Quadriacanthus Paperna, 1961 (Monogenea: Ancyrocephalidae). The following parasites are described: Quadriacanthus longifilisi n. sp., Q. thysi n. sp., Q. ayameensis n. sp., Q. agnebiensis n. sp., Q. simplex n. sp., Q. gourenei n. sp. and Q. macrocirrus n. sp.

Animals↗
PubMed1999

Socioeconomic and psychosocial conditions of parents and cardiovascular risk factors in their children: the Belgian Luxembourg Child Study III.

Socioeconomic and psychosocial handicaps are often associated with disease. There is a large body of information on adults on such relationships, but data are sparse on children. In a cohort of 1028 boys and girls, selected at random from school classes in Province de Luxembourg, a mainly rural area of Belgium, these problems were analysed in age strata of 6-8, 8-10, 10-12 years. Participation rate was 71%. Information was collected from questionnaires,. Anthropometric variables, blood pressure and glucose as well as cholesterol, triglycerides and insulin were measured in the children and related to the psychosocial and socioeconomic factors, organized into rural, psychosocial, socioeconomic and alcohol/smoking clusters of observations. Rural: Duration of living in the area of the child and parents correlated with diastolic blood pressure, particularly in boys (p < 0.01). Psychosocial: Housewives (p = 0.002) and their children (p = 0.002) had higher body mass indexes (BMI) than other mothers and their children. Sons of housewives also had higher blood pressure (systolic, p = 0.0007, diastolic, p = 0.007). Socioeconomic: Socioeconomic factors of parents (profession, unemployment) played relatively minor roles. Alcohol/smoking: Alcohol consumption was related to skinfold thickness in boys (p = 0.022), but not in girls. Girls, but not boys, with smoking parents had higher BMI (p=0.014). Multiple regression analyses suggested that psychosocial factors, such as housewives as mothers of large families, may be important for associations with cardiovascular risk factors in their children. There were apparent differences in the findings between girls and boys, suggesting that boys are more vulnerable to the impacts of the factors analysed.

Adult↗
PubMed1999

Ovulation induction with low dose alternate day recombinant follicle stimulating hormone (Puregon).

We investigated whether a recombinant follicle stimulating hormone (FSH) (Puregon) can be administered less frequently and at lower doses during ovulation induction than is current practice. Patients (20-35 years, body mass index <30 kg/m(2)) with infertility and chronic anovulation secondary to polycystic ovarian syndrome and resistant to previous clomiphene treatment received (Puregon); 100 IU, n = 17 patients, or 50 IU, n = 10 patients) on alternate days. After 2 weeks and in the absence of follicular recruitment, doses were increased stepwise at weekly intervals (50 IU/alternate days). Twenty-two cycles out of 27 were ovulatory. There were six pregnancies, five from Puregon (100 IU) and one from Puregon (50 IU); four pregnancies proceeded to term. The duration of stimulation (mean, range) with Puregon (100 IU) was 16.4, 7-29 and Puregon (50 IU) 19.1, 8-38 days. The gonadotrophin doses administered (mean; range) were 689, 200-1800 IU (Puregon 50 IU) and 939, 400-2300 IU (Puregon 100 IU). We conclude that low dose alternate day Puregon treatment is suitable for this difficult patient group.

Adult↗
PubMed1999

Vitamin D regulates human ectocervical epithelial cell proliferation and insulin-like growth factor-binding protein-3 level.

The differentiation status of the cervical epithelial cell has an important influence on responsiveness to estrogens and progestins. Several agents, including glucocorticoids and retinoids, are known to influence cervical cell differentiation. However, the effects of vitamin D have not been examined. Vitamin D is known to regulate cell proliferation and gene expression in a variety of epithelial cells. In the present study we investigated the ability of 1alpha25-dihydroxyvitamin D3 (D3) to regulate cell proliferation and expression of insulin-like growth factor-binding protein-3 (IGFBP-3) in human ectocervical epithelial cells. ECE16-1, a non-tumorigenic cervical cell line, was growth inhibited by D3 with maximal inhibition at 1000 nM. IGFBP-3 levels increased in parallel with the growth inhibition. IGFBP-3 levels were half-maximally increased at approximately 10-100 nM and maximally increased (10- to 30-fold) at 1000 nM D3. These studies show that vitamin D regulates cervical epithelial cell gene regulation and cell proliferation and that IGFBP-3 may be an in vivo marker of vitamin D action in the cervix.

Calcitriol↗
PubMed1999

Insulin-like growth factor binding protein-3 does not mediate the interferon-dependent suppression of human ectocervical epithelial cell proliferation.

Interferon is a potential therapeutic agent for the treatment of cervical cancer. In the present study we examine the role of IFNgamma as a regulator of proliferation and production of IGFBP-3 expression in ectocervical epithelial cells. ECE16-1 cells are a model for studying early human papillomavirus-dependent cervical disease. IFNgamma produces a concentration-dependent inhibition of ECE16-1 cell proliferation that is associated with an increase in insulin-like growth factor binding protein-3 level. Growth suppression and IGFBP-3 increase is maximal at concentrations of IFNgamma >/=0.75 ng/ml. The increased IGFBP-3 expression is mediated via an increase in IGFBP-3 encoding mRNA. In contrast, IFNgamma inhibits proliferation of CaSki and SiHa cells, but IGFBP-3 is barely detectable and levels are not regulated by IFNgamma. These results suggest that the IFNgamma-dependent suppression of CaSki and SiHa cell proliferation is not mediated by secreted IGFBP-3. This result was confirmed when vector-mediated overexpression of immunoreactive IGFBP-3 in SiHa and CaSki cells did not consistently result in reduced cell proliferation rate.

Animals↗
PubMed1998

Cellular expression of growth hormone and prolactin receptors in human breast disorders.

Growth hormone (GH) and prolactin (PRL) exert their regulatory functions in the mammary gland by acting on specific receptors. Using isotopic in situ hybridization and immunohistochemistry, we have localized the expression of hGH receptor (hGHR) and hPRL receptor (hPRLR) in a panel of human breast disorders. Surgical specimens from adult females included normal breast, inflammatory lesions (mastitis) benign proliferative breast disease (fibroadenoma, papilloma, adenosis, epitheliosis), intraductal carcinoma or lobular carcinoma in situ, and invasive ductal, lobular or medullary carcinoma. Cases of male breast enlargement (gynecomastia) were also studied. In situ hybridization analysis demonstrated the co-expression of hGHR and hPRLR mRNA in all samples tested. Epithelial cells of both normal and tumor tissues were labelled. Quantitative estimation of receptor mRNA levels was regionally measured in areas corresponding to tumor cells and adipose cells from the same section. It demonstrated large individual variation and no correlation emerged according to the histological type of lesion. Receptor immunoreactivity was detected both in the cytoplasm and nuclei or in the cytoplasm alone. Scattered stromal cells were found positive in some cases, but the labeling intensity was always weaker than for neoplastic epithelial cells. Our results demonstrate the expression of the hGHR and hPRLR genes and their translation in epithelial cells of normal, proliferative and neoplastic lesions of the breast. They also demonstrate that stromal components express GHR and PRLR genes. Thus the putative role of hGH or hPRL in the progression of proliferative mammary disorders is not due to grossly altered levels of receptor expression.

Adult↗
PubMed1998

Obesity and nutrition in children. The Belgian Luxembourg Child Study IV.

OBJECTIVE: To analyse the association between nutritional and familial factors and obesity in boys and girls. DESIGN: Randomized, cross-sectional population study. SETTING: Province de Luxembourg, Belgium. SUBJECTS: One thousand and twenty-eight boys and girls in age strata 6-8, 8-10 and 10-12 y, comprising 70.3% of primary cohort. METHODS: Examinations included anthropometric measurements and questionnaires covering familial, socioeconomic and psychosocial factors. A three day dietary record was obtained in 955 children. This was analysed in relation to the anthropometric data. RESULTS: In comparison with similar studies from other regions and recommended allowances, the intakes of total energy, fat, particularly saturated fat and cholesterol, were high, while consumption of carbohydrate and fiber was low, as well as the polyunsaturated/saturated ratio of fat. Total energy intake showed no or weakly significant correlations with anthropometric factors. However, total fat (P=0.045) and saturated fat (P=0.0005) intake showed consistent positive correlations with body mass index (BMI, kg/m2) and skinfold thickness, with corresponding negative relationships to carbohydrate intake (P=0.034) in boys. Such relationships were also found when calculated as energy density. These associations were not statistically significant in girls. The high fat, low carbohydrate pattern of the nutritional status seemed to be more pronounced in families where the father had a low level of education (lipids, boys, P=0.0007), and where both parents were obese (saturated fat, boys, P=0.023), suggesting involvement of socioeconomic and familial factors. CONCLUSION: The lack of correlation between factors indicating obesity and total energy intake suggests that the positive energy balance causing obesity is due mainly to a low energy output. However, since energy intake measurements are imprecise, overeating can not be excluded, particularly since elevated consumption of food with high contents of fat, found in these children seems to be poorly regulated.

Belgium↗
PubMed1998

A rapid method for the measurement of the oxoreductase activity of 11beta-hydroxysteroid dehydrogenase in granulosa-lutein cells from patients undergoing in-vitro fertilization.

The presence of 11beta-hydroxysteroid dehydrogenase (11beta-HSD; EC1.1.1.146), the enzyme responsible for the interconversion of cortisol and cortisone, in granulosa-lutein (GL) cells is associated with a poor outcome in in-vitro fertilization (IVF). We have developed a simple method of assessing the reductase component of 11beta-HSD in these cells which is sufficiently rapid to provide data on the enzyme's activity prior to embryo replacement. Cells were pooled from follicular aspirates and challenged with cortisone within 2 h of aspiration. Cortisol secretion was then measured by radioimmunoassay. Conversion of cortisone to cortisol was linear for up to 3 h and was completely inhibited by glycyrrhetinic acid, a specific 11beta-HSD inhibitor. Initial velocity rates were determined for eight cortisone concentrations (range 0.1-8 micromol/l), and the apparent Km calculated (1.6 +/- 0.4 micromol/l). There was no evidence of substrate/product inhibition and conversion of cortisone to cortisol was <2% in all experiments. In subsequent work, cells were challenged with cortisone (6 micromol/l) for 2 h. Cells challenged for 2 h immediately following purification from follicular aspirates produced varying amounts of cortisol (range 25-150 nmol/pooled follicles from each patient, n = 10 patients), while basal outputs were <6 nmol/l. Enzyme activity was also examined in cells on a per follicle basis from individual patients and found to vary considerably (e.g. 19, 53 and 36 nmol/l cortisol/1000 cells, three follicles). Having established the method for assessing 11beta-reductase activity within GL cells, we performed a small prospective study on a series of 20 patients examining the enzyme activity within 110 individual follicles. 11Beta-reductase activity varied greatly from patient to patient and from follicle to follicle ranging from <0.024-0.57 nmol cortisol/microg DNA but at present low patient numbers preclude a meaningful correlation between enzyme activity and pregnancy rate. In summary, we have developed a simple, rapid (<8 h) assay for detecting the reductase activity of 11beta-HSD in GL cells isolated from pooled or individual follicles. This procedure is sufficiently quick to aid in the choice of embryo for replacement.

11-beta-Hydroxysteroid Dehydrogenases↗
PubMed1998

Gonadotrophin heterogeneity and biopotency: implications for assisted reproduction.

The extensive heterogeneity of the gonadotrophin hormones, follicle stimulating hormone (FSH) and luteinizing hormone (LH), is due primarily to the heterogeneous nature of their carbohydrate side-chains, in particular sialic acid residues. In this review, we discuss the role of carbohydrate chains in receptor binding and activation, biological activity, and metabolic half-life. The synthesis and secretion of the various glycoforms of both FSH and LH appear to be under endocrine control with gonadotrophin-releasing hormone (GnRH), oestradiol and testosterone playing important roles. Evidence for different glycoforms having variable biopotency or different encoded functions is increasing, and the production and secretion of more or less acidic gonadotrophin species in different physiological states may represent an important mechanism whereby the pituitary regulates gonadal cell and organ function. This has potential importance for the development of new pharmaceutical reagents and new therapeutic regimens in assisted reproduction. It is envisaged that the use of existing and new forms of FSH/LH will allow patients to be treated in a more controlled and physiological manner, with treatment regimens individualized to the needs of the patient.

Animals↗
PubMed1998

In vitro cross-linking of recombinant human involucrin.

Human involucrin (hINV) is a constituent of the scaffolding of the cornified envelope. In the present study, we describe an in vitro model system to study the role of hINV in scaffold formation. We characterize the in vitro cross-linking of full-length (585 amino acid) recombinant hINV, rhINV(1-585). When reacted with detergent-solubilized, particulate transglutaminase type 1 (TG1) or partially purified type 2 transglutaminase (TG2), rhINV(1-585) functions as a TG substrate in a calcium-dependent manner. When the reaction is supplemented with 14C-putrescine tracer, the radiolabeled cosubstrate is incorporated into a high-molecular-weight product in a calcium-, rhINV(1-585)- and time-dependent manner. 35S-rhINV(1-585) is also cross-linked to form a high-molecular-weight product. These results suggest that rhINV(1-585) is extensively multimerized. Products having a molecular weight smaller than authentic rhINV(1-585) are also formed, providing evidence for intramolecular cross-link formation. Transmission electron microscopy of cross-linked product reveals immunoreactive large-molecular-weight loop-string-loop and branched structures. Our studies (1) show that rhINV(1-585) is a substrate for both TG1 and TG2, (2) indicate that rhINV(1-585) can be cross-linked to form macromolecular products having distinct structural features, (3) demonstrate that rhINV(1-585) forms intramolecular cross-links when hINV concentration is limiting and (4) establish that hINV possesses reactive Gln and Lys residues.

Bacteria↗
PubMed1998

Stimulatory effect of glucose on macrophage lipoprotein lipase expression and production.

Cardiovascular diseases are the leading cause of morbidity and mortality in diabetes. Lipoprotein lipase (LPL), a major secretory product of macrophages, has been suggested to play a key role in the development of atherosclerosis. In the present study, we evaluated the effect of high glucose on macrophage LPL mRNA expression and secretion. Exposure of murine J774 macrophages to high D-glucose concentrations (20-30 mmol/l) resulted in a dramatic upregulation of LPL mRNA expression and immunoreactive mass. This effect was not observed when these cells were incubated in the presence of L-glucose or mannitol. High glucose concentrations were also found to enhance LPL gene expression and immunoreactive mass in human monocyte-derived macrophages. J774 cells cultured in a high glucose environment expressed increased c-fos mRNA levels. Treatment of these cells with c-fos antisense DNA or protein kinase C inhibitor inhibited the stimulatory effect of glucose on LPL mRNA expression. In J774 cells exposed to high glucose concentrations, enhanced nuclear protein binding to the AP-1-responsive region of the murine LPL promoter was observed, while LPL mRNA stability remained unchanged. Overall, these results demonstrate that high glucose upregulates macrophage LPL gene expression and immunoreactive mass and that this effect involves transcriptional events.

Animals↗
PubMed1997

Physical activity, obesity, and cardiovascular risk factors in children. The Belgian Luxembourg Child Study II.

Physical activity was measured in relation to cardiovascular (CV) risk factors in a randomly selected population of 1028 children from Province de Luxembourg in Belgium, a mainly rural area with a high prevalence of such risk factors among adults and children. Physical activity was estimated as participation in sport activities, a major indicator of leisure-time physical activity in schoolchildren, and physical inactivity was estimated as frequency and duration of television (TV) watching. Boys participated more frequently in sport activities than girls did (p = 0.001). A majority of the children watched TV daily. After age adjustment, bodyweight (girls, p < 0.012; boys, p < 0.027) and, in boys, body mass index (BMI) (p < 0.039) were related to days per week of TV watching. No significant relationships with other CV risk factors remained after adjustments for BMI. In analyses of independent contributions of age, TV watching, and sports activity on CV risk factors, age showed highly significant relationships. In boys, TV showed relationships with BMI (p < 0.04) and (borderline) with systolic blood pressure, independent of age and sports activity, whereas the latter was significantly related to subscapular skinfold (p < 0.04) and (borderline) with triceps skinfold and cholesterol. In girls, no significant independent contributions to risk factor associations were found. The father's education was directly associated with sports activities, whereas the mother being a housewife showed negative relationships to physical activity and positive to TV watching in their children, suggesting socioeconomic influence on the activity patterns of children. Furthermore, registrations suggested less physical activity in the most rural part of the area. It is concluded that children in this mainly rural area watch TV frequently. In boys, physical inactivity, measured both as TV watching and as registrations of sports activities, contributes independently to body fat mass. In girls, no contribution or weaker contributions of physical inactivity were found. This suggests that contributory factors leading to obesity might be different in girls and boys.

Belgium↗