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C Jean

Publications and source records attributed to C Jean.

At least 37 records · Page 2Linked to original sources
PubMed1997

Androgen induction of the SVS family related protein MSVSP99: identification of a functional androgen response element.

The gene encoding MSVSP99 (mouse seminal vesicle secretory protein of 99 amino acids) is specifically expressed in the mouse seminal vesicle under androgenic control. To study hormonal regulation, fragments of the 5'-flanking region, extending from -2365 to +16 were linked to the chloramphenicol acetyl transferase (CAT) gene and cotransfected with an androgen receptor expression vector into CV-1 cells. A minimal region (-387 to +16) was sufficient for full androgen induction. Further deletion, up to nt-261, almost completely abolished androgen inducibility. DNase I footprinting and band-shift assays, using the DNA binding domain of the androgen receptor (AR-DBD), revealed three AR binding sites: two putative androgen response elements (AREs) occurring at positions -361 (AREd) and -208 (AREp), and an androgen receptor binding region (ARBR) located between positions -317 and -293. Transient transfection assays revealed that site-directed mutation in AREp abolished androgen induced expression, whereas mutation in AREd or in ARBR had no effect. The results demonstrate that AREp is a functional sequence that must cooperate with additional cis-acting elements, located between -387 and -261, for androgen induction of the MSVSP99 gene.

Animals↗
PubMed1997

Isolation of the mouse aldose reductase promoter and identification of a tonicity-responsive element.

Aldose reductase (AR; EC 1.1.1.21) is an oxidoreductase that catalyzes the NADPH-dependent conversion of glucose to sorbitol, the first step of the polyol pathway. AR is of great interest due to its implication in the etiology of diabetic complications. In renal medullary cells, AR also plays an osmoregulatory role by accumulating sorbitol to maintain the intracellular osmotic balance during antidiuresis. We have previously cloned the AR cDNA from mouse kidney, and we report here the isolation of the mouse AR gene promoter. Transient transfection of chloramphenicol acetyltransferase reporter constructs containing various 5'-flanking regions of the mouse AR gene in CV1 cells led to the identification of a sequence spanning base pairs -1053 to -1040, required for an enhancer activity in hypertonic compared with isotonic cell culture conditions. This sequence is similar to the tonicity-responsive element first characterized in the betaine-gamma-aminobutyric acid transporter promoter.

Aldehyde Reductase↗
PubMed1997

The androgen-dependent mouse seminal vesicle secretory protein of 99 amino acids (MSVSP99): regulation of the mRNA and preliminary characterization of the promoter.

MSVSP99 (mouse seminal vesicle secretory protein of 99 amino acids) is a member of the rat and mouse seminal vesicle secretory protein (SVS) family. In order to characterize its androgenic regulation, the cloned cDNA and gene encoding MSVSP99 have been used. At adulthood, the MSVSP99 mRNA represents from 3 to 7% of the total mRNA population. This mRNA accumulation is under androgenic control because it is abolished by castration and restored in castrated mice by heptylate testosterone injection. During ontogenesis, MSVSP99 mRNA is just detectable in 10-day-old mice, and reaches adult levels at 30 days. Neonatal castration abolishes MSVSP99 mRNA accumulation in 20-day-old mice. Transcription elongation assays show that androgens act mainly on the MSVSP99 gene transcription. In an attempt to obtain information about the mechanism of androgen action on transcription, preliminary transient transfection experiments in CV-1 cells permitted us to define a promoter region (-387/ + 16), the activity of which is enhanced by dihydrotestosterone.

Androgens↗
PubMed1997

Immunological detection of potential signal-transduction proteins expressed during wheat somatic tissue culture.

An immunochemical approach was used to detect the expression of putative guanine nucleotide-binding proteins (G-proteins), arrestin, and nucleoside diphosphate kinases during wheat (Triticum aestivum) tissue culture initiated from immature embryos. Both the soluble and membrane extracts from the immature embryos revealed bands of 58, 40, and 16 kD with antibodies to G-protein (alpha subunit), arrestin, and nucleoside diphosphate kinase, respectively. These proteins were overexpressed in vitro in both nonembryogenic callus and embryogenic cultures. An additional soluble protein (32 kD) was detected by anti-G alpha antibodies in cultured tissues but not in immature embryos, suggesting a possible function in cell multiplication. Moreover, somatic embryogenesis was associated with the appearance of a 29-kD protein reactive with anti-arrstin antibodies, both in soluble and membrane fractions. Tissue-cultured genetic stocks of Chinese Spring wheat, including the disomic, 36 ditelosomic, and 6 nullisomic-tetrasomic wheat lines, were used to ascertain the chromosomal location of the genes encoding the 29-kD arrestin-like protein. The lack of a signal with the nonembryogenic ditelosomic 3 D short chromosome arm line suggests that the 3 D long chromosome arm possesses at least one gene involved in the expression of the 29-kD protein. The putative role of the 29-kD protein in signal-transduction regulating embryogenesis is discussed.

Amino Acid Sequence↗
PubMed1996

Protein kinase C pathway potentiates androgen-mediated gene expression of the mouse vas deferens specific aldose reductase-like protein (MVDP).

Transcription of the mouse vas deferens protein (MVDP) gene, a member of the aldo-keto reductase superfamily, is stimulated by androgens via the androgen responsive element (ARE) located in the proximal promoter (-111 to -97). We investigated interaction between androgens and the protein kinase C (PKC) signalling pathway. Transcriptional regulation was determined by analysis of chloramphenicol acetyltransferase (CAT). T47D cells were transiently transfected with 5' flanking MVDP DNA promoter sequences (-1804 to +41; -510 to +41 and -121 to +41) fused to the reporter (CAT) gene. Androgen-induced transcriptional activity can be enhanced from 6 (1.8 and 0.5 kb MVDP-CAT constructs) to 18 fold (0.16 kb MVDP-CAT construct), in a time and dose-dependent manner, by the PKC activator 12-o-tetradecanoylphorbol-13 acetate (TPA). A mutation in the proximal ARE abolished both androgen and TPA-dependent gene enhancement. TPA influenced minimally MMTV promoter in T47D cells and MVDP promoter in CV1 cells suggesting that the effects of the PKC activator are probably promoter and cell-specific. In contrast, activation of protein kinase A (PKA) via addition of dibutyryl-cAMP (db-cAMP) reduced androgen induction of the MVDP gene.

Aldehyde Reductase↗
PubMed1996

Face-straight-down and face-near-straight-down positions in healthy, prone-sleeping infants.

OBJECTIVE: To determine the frequency and physiologic consequences of the face-straight-down (FSD) position, a postulated mechanism for the sudden infant death syndrome in prone-sleeping infants. STUDY DESIGN: A survey of 151 infants, aged 1 to 7 months, in Montreal showed that 33% slept prone. Ten healthy prone-sleeping infants were studied in their homes at age 10 to 22 weeks. Infrared video and cardiorespiratory recordings were made on 3 consecutive nights in the prone (nights 1 and 3) and lateral (night 2) positions. RESULTS: Infants maintained the prone position during 17 of 19 studies, but only 4 of 9 infants maintained the lateral position. The FSD position was observed 27 times in 17 prone nights: median frequency, 0.6 times per night (interquartile range, 0 to 4), and median total duration, 3.3 minutes (0.8% of total sleep time). A related position, the face-near-straight-down (FNSD) position, occurred more often, 5.3 (1 to 10) time per prone night, for 22.4 minutes (5.8% of total sleep time). Most periods in the FSD and FNSD position had no physiologic consequences; however, 14% of FSD and 3% of FNSD episodes were associated with airway obstruction as indicated by snoring, paradoxical respiratory movements, apnea, and/or increased partial pressure of transcutaneous carbon dioxide. Spontaneous arousal and head turning terminated the FSD and FNSD episodes. CONCLUSION: The FSD and FNSD positions occur commonly in healthy prone-sleeping infants, and these positions can cause airway obstruction. We speculate that those infants with sudden infant death syndrome found in the FSD or FNSD position either have a congenital or an acquired defect in the arousal-head turning response or have encountered insurmountable environmental factors that prevent effective head turning.

Airway Obstruction↗
PubMed1996

Comparative efficacy and safety of single oral doses of sparfloxacin versus ciprofloxacin in the treatment of acute gonococcal urethritis in men.

A double-blind, randomised, international multicentre study was conducted to compare the efficacy and safety of sparfloxacin 200 mg versus ciprofloxacin 250 mg as single oral-dose treatment for acute gonorrhoea in men. A total of 238 patients were included and 191 were evaluable for primary efficacy (eradication of Neisseria gonorrhoeae). At follow-up, 99% (96/97) of the sparfloxacin-treated patients were culture-negative compared with 98% (92/94) in the ciprofloxacin group. Three of the four patients who harboured gonococci at follow-up admitted to having had unprotected sexual intercourse after treatment and were probably reinfections. The rate of post-gonococcal urethritis was 26% in both groups, although Chlamydia trachomatis was isolated in only 4% of patients at inclusion. Both drugs were well tolerated. Single oral doses of sparfloxacin 200 mg and ciprofloxacin 250 mg are equally effective in the treatment of acute gonorrhoea in men.

Acute Disease↗
PubMed1996

A single gamma-tubulin gene and mRNA, but two gamma-tubulin polypeptides differing by their binding to the spindle pole organizing centres.

Cells of eukaryotic organisms exhibit microtubules with various functions during the different developmental stages. The identification of multiple forms of alpha- and beta-tubulins had raised the question of their possible physiological roles. In the myxomycete Physarum polycephalum a complex polymorphism for alpha- and beta-tubulins has been correlated with a specific developmental expression pattern. Here, we have investigated the potential heterogeneity of gamma-tubulin in this organism. A single gene, with 3 introns and 4 exons, and a single mRNA coding for gamma-tubulin were detected. They coded for a polypeptide of 454 amino acids, with a predicted molecular mass of 50,674, which presented 64-76% identity with other gamma-tubulins. However, immunological studies identified two gamma-tubulin polypeptides, both present in the two developmental stages of the organism, uninucleate amoebae and multinucleate plasmodia. The two gamma-tubulins, called gamma s- and gamma f-tubulin for slow and fast electrophoretic mobility, exhibited apparent molecular masses of 52,000 and 50,000, respectively. They were recognized by two antibodies (R70 and JH46) raised against two distinct conserved sequences of gamma-tubulins. They were present both in the preparations of amoebal centrosomes possessing two centrioles and in the preparations of plasmodial nuclear metaphases devoid of structurally distinct polar structures. These two gamma-tubulins exhibited different sedimentation properties as shown by ultracentrifugation and sedimentation in sucrose gradients. Moreover, gamma s-tubulin was tightly bound to microtubule organizing centers (MTOCs) while gamma f-tubulin was loosely associated with these structures. This first demonstration of the presence of two gamma-tubulins with distinct properties in the same MTOC suggests a more complex physiological role than previously assumed.

Amino Acid Sequence↗
PubMed1996

Oral empiric treatment of community-acquired pneumonia. A multicenter, double-blind, randomized study comparing sparfloxacin with roxithromycin. The Scandinavian Sparfloxacin Study Group.

STUDY OBJECTIVE: Comparison of efficacy and safety of sparfloxacin (Spfx) vs roxithromycin (ROXI) for treatment of community-acquired pneumonia (CAP). DESIGN: Multicenter, double-blind, randomized study. SETTING: Twenty-three university and community hospitals in Scandinavia. PATIENTS: Three hundred four adults (> or = 18 years of age) with CAP treated as outpatients (25%) or inpatients (75%). INTERVENTIONS: Randomization 1:1 to Spfx, 400 mg on day 1, then 200 mg once daily, or ROXI, 150 mg twice daily, 10 to 14 days. Safety and efficacy analyses in intention-to-treat (ITT) and evaluable populations. RESULTS: Three hundred three of 304 patients were included in the ITT and safety analyses and 260 (86%) were evaluable at the end of follow-up. Streptococcus pneumoniae was the cause of pneumonia in 62 (20%) patients (11 with bacteremia), Chlamydia pneumoniae in 40 (13%), and Mycoplasma pneumoniae in 38 (13%) patients. The success rates for Spfx and ROXI at the end of follow-up were 82% and 72%, respectively, in the ITT population, and 94% and 79%, respectively, in the evaluable population. The odds ratio Spfx/ROXI for success was 4.5 (95% confidence interval, 1.9, 10.8) for the evaluable population. Both drugs were, overall, equally safe. GI symptoms were the most common adverse experiences in both groups. Prolongation of QTc, without clinical symptoms, was seen in 3% of Spfx patients and in 1% of ROXI patients, and photosensitivity, mostly mild to moderate, was seen in 5% of the Spfx group. CONCLUSIONS: Oral treatment with Spfx was superior to ROXI for the treatment of moderately severe CAP. Spfx was effective for all isolated pathogens, including S pneumoniae, and may be an alternative for empiric treatment of CAP, especially in areas with a high incidence of beta-lactam-resistant pneumococci.

Administration, Oral↗
PubMed1995

Characterization of the promoter of the gene for a mouse vas deferens protein related to the aldo-keto reductase superfamily: effect of steroid hormones and phorbol esters.

Mouse vas deferens protein (MVDP) is a member of the aldo-keto reductase family regulated by androgens. The expression of a hybrid gene containing the promoter of the MVDP gene and the chloramphenicol acetyl transferase (CAT) gene coding region was analyzed in two cell lines that do not normally express the MVDP gene: T47D and CV1 cells. A small region of the promoter (-121 to +41) was able to direct significant expression of the reporter gene in both cell lines. Additional elements, between -510 and -121 modulate basal expression in a cell-dependent manner. Interestingly, the 162 bp fragment serves as an androgen-dependent enhancer, and mutation of the consensus ARE sequence located between positions -111 and -97 resulted in a loss of androgen response in both cell lines. Additional elements, upstream of the enhancer, modulate induction positively or negatively in relation to the cell line used. The expression of different MVDP-CAT constructs was more effectively induced by androgens than by glucocorticoids at physiological hormonal concentrations. In addition to the 162 bp enhancer, sequences upstream of -510 were also required for specific androgen regulation. Phorbol 12-myristate 13-acetate (TPA) had no effect on basal activity of the 1.8 kb MVDP-CAT construct but strongly enhanced the induction by androgens.

Alcohol Oxidoreductases↗
PubMed1995

Exportation of mouse vas deferens protein, a protein without a signal peptide, from mouse vas deferens epithelium: a model of apocrine secretion.

Mouse vas deferens protein (MVDP) is a major androgen-dependent protein of deferential fluid. It is specifically expressed in the epithelium of the mouse vas deferens. Its amino acid sequence as deduced from the nucleotidic sequence of its cDNA does not possess a signal sequence characteristic of secretory proteins. In vitro, transcription of MVDP cDNA followed by translation of mRNA in the rabbit reticulocyte system, in the absence or the presence of microsomes, demonstrated that there was no internalization of MVDP into microsomes that could protect it from degradation by proteinase K; this confirmed the absence of signal sequence. Moreover, MVDP has its NH2-terminus blocked. To understand how MVDP can be exported, its ultrastructural distribution and secretion process were analyzed by means of electron microscopy. Immunolocalization of MVDP revealed that it was distributed in the whole cytoplasm; it was never detected in the lumen of endoplasmic reticulum, Golgi apparatus, or vesicles but was abundant in apical protrusions and in the fluid, where it was associated with cellular material undergoing degradation. These data clearly demonstrated that exportation of MVDP into the luminal fluid does not occur in the classical manner for secretory proteins but rather involves an apocrine secretion process.

Aldehyde Reductase↗
PubMed1995

Structure and sequence of a mouse gene encoding an androgen-regulated protein: a new member of the seminal vesicle secretory protein family.

The gene encoding MSVSP99 (mouse seminal vesicle secretory protein of 99 amino acids), an androgen-dependent protein specifically expressed in the mouse seminal vesicle, was isolated and sequenced. A mouse genomic library constructed in the lambda EMBL12 vector was screened using a full length cDNA probe. One genomic clone was selected, 7.4 kb of which were shown to contain the whole MSVSP99 gene. The complete sequence of the MSVSP99 gene (1.7 kb), plus 0.8 and 0.3 kb of the 5' and 3' flanking regions respectively, has been determined. The gene is composed of four exons interrupted by three introns. The size range for the four exons is 47-217 bp, while that of introns is 87-615 bp. The transcription start site was identified as an adenine residue located 21 nucleotides upstream from the ATG start codon. Putative TATA and CAAT boxes were identified, along with a number of regions that shared homologies with known regulatory sequences. These included androgen-responsive elements located in the promoter as well as in the gene sequence. Sequence comparisons with other androgen-responsive genes showed strong homologies between the MSVSP99 gene and the seminal vesicle secretory protein (SVS) family genes (rat SVS II, IV, V and VI). Moreover, some regions were found to be conserved between the MSVSP99 gene and the human semenogelin I and II genes.

Amino Acid Sequence↗
PubMed1994

Identification of a functional androgen response element in the promoter of the gene for the androgen-regulated aldose reductase-like protein specific to the mouse vas deferens.

Mouse vas deferens protein (MVDP), a member of the aldo-keto reductase superfamily, is exclusively produced in the epithelial cells of the deferent duct under androgenic regulation. To better understand androgenregulated MVDP gene expression, the location and sequences of androgen response elements (AREs) in the 5'-flanking DNA were determined. Sequence analysis revealed two putative AREs as follows: one between positions -1186 and -1171 (distal ARE) and the other between -111 and -97 (proximal ARE). To study hormonal regulation, fragments of the MVDP promoter region, extending from residue -1804 to +41, were linked to the chloramphenicol acetyltransferase (CAT) reporter gene and cotransfected with a human androgen receptor expression vector into T47D cells in a transient expression assay. A minimal region (-121 to +41) was identified as being sufficient for androgen-regulated gene expression. A mutation in proximal ARE almost completely abolished androgen induction of CAT. One copy of the sequence TGAAGT tcc TGTTCT, cloned in the opposite orientation in front of the thymidine kinase promoter, confers androgen responsiveness to the CAT reporter gene. Androgen transcriptional activity was not detected with the distal ARE. The data provide strong evidence that transcriptional regulation of the MVDP gene occurs via the sequence TGAAGT tcc TGTTCT.

Aldehyde Reductase↗
PubMed1994

Interaction of G-actin with thymosin beta 4 and its variants thymosin beta 9 and thymosin beta met9.

Thymosin beta 4 is a major actin sequestering peptide in vertebrate cells and plays a role in the regulation of actin monomer/polymer ratio. Thymosin beta 9 and thymosin beta met9 are minor variants of thymosin beta 4. The possible function of these peptides has been investigated by comparing the actin binding properties of these beta-thymosins. Thymosin beta 9 and thymosin beta met9 were found to inhibit polymerization of ATP-actin with identical KDs of 0.7-0.8 microM (as compared to 2 +/- 0.3 microM for thymosin beta 4); like thymosin beta 4, they bound to ADP-G-actin with a 100-fold lower affinity than to ATP-G-actin. The interaction of thymosin beta 4 and thymosin beta met9 with G-actin was weakened 20-fold upon oxidation of methionine-6 into methionine sulfoxide. Binding of thymosin beta 4 to G-actin was accompanied by a 15% increase in the fluorescence intensity of actin tryptophans, and a 10 nm emission blue shift. Methionine-6 played an important role in this effect. The fluorescence change was used to monitor the kinetics of thymosin beta 4 binding to G-actin in the stopped-flow. The reaction was bimolecular, with association and dissociation rate constants of approximately 1.5 microM-1 s-1 and 2 s-1 respectively, under physiological conditions. The possible physiological significances of methionine-6 oxidation and of the relatively slow binding kinetics in regulating thymosin beta 4 function in vivo is discussed.

Actins↗
PubMed1994

Androgens regulate expression of the gene coding for a mouse vas deferens protein related to the aldo-keto reductase superfamily in epithelial cell subcultures.

Mouse vas deferens protein (MVDP), a member of the aldo-keto reductase superfamily, is exclusively produced in the vas deferens. To better understand androgen-regulated MVDP gene expression we have used RNA hybridization to study the effects of androgens on the steady-state levels of MVDP mRNA in vas deferens epithelial cell subcultures. Northern blot analysis revealed that these cells only express MVDP mRNA in the presence of androgens. There was a close relationship between MVDP mRNA levels and dihydrotestosterone concentrations. MVDP mRNA is induced over a period of 24h and maximal induction is about 25-fold. Treatment of cells with cycloheximide completely abolished the observed androgen effect suggesting that the induction of the MVDP gene by androgens depends on continuous protein synthesis. Transient transfection of vas deferens epithelial cells with MMTV-CAT vector showed that these cells contained functional androgen receptors and that they are a suitable system to study androgen effect on MVDP gene regulatory elements.

3T3 Cells↗
PubMed1994

Diminished diabetogenic effect of streptozotocin in adrenodemedullated rats.

Although stress is suspected to play a role in the development of diabetes mellitus, no direct experimental evidence for involvement of the adrenal medullae in onset of the disease has yet been found. We recently observed that, in comparison with sham-operated rats, fewer adrenodemedullated rats become diabetic after an i.v. injection of streptozotocin (STZ); thus, the present study examined the role of the adrenal medullae in the development of experimental diabetes. Male Wistar rats were surgically adrenodemedullated (ADM) or sham-operated (SHAM). After a 3-week recovery period, they were injected with 40 mg STZ/kg freshly dissolved in citrate buffer or buffer alone. A 1-mL arterial blood sample was withdrawn 12 days later in previously cannulated animals; then the rats were killed and their pancreas was removed. The plasma glucose levels were lower in ADM rats injected with buffer alone than in their SHAM counterparts (7.7 +/- 0.1 vs. 8.5 +/- 0.1 mmol/L; p < 0.05). The glucose levels were higher (p < 0.001) in both groups of STZ-treated rats, with values 28% lower in ADM than in SHAM rats (14.7 +/- 1.6 vs. 18.8 +/- 1.2 mmol/L; p < 0.03). Whereas plasma insulin levels did not differ between ADM and SHAM rats injected with buffer alone (431 +/- 38 vs. 428 +/- 35 pmol/L; p > 0.05), they were diminished in SHAM animals injected with STZ (292 +/- 37 vs. 428 +/- 35 pmol/L; p < 0.02).(ABSTRACT TRUNCATED AT 250 WORDS)

Adrenal Medulla↗
PubMed1993

Modulation of the interaction between G-actin and thymosin beta 4 by the ATP/ADP ratio: possible implication in the regulation of actin dynamics.

The interaction of G-actin with thymosin beta 4 (T beta 4), the major G-actin-sequestering protein in motile and proliferating cells, has been analyzed in vitro. T beta 4 is found to have a 50-fold higher affinity for MgATP-actin than for MgADP-actin. These results imply that in resting platelets and neutrophils, actin is sequestered by T beta 4 as MgATP-G-actin. Kinetic experiments and theoretical calculations demonstrate that this ATP/ADP dependence of T beta 4 affinity for G-actin can generate a mechanism of desequestration of G-actin by ADP, in the presence of physiological concentrations of T beta 4 (approximately 0.1 mM). The desequestration of G-actin by ADP is kinetically enhanced by profilin, which accelerates the dissociation of ATP from G-actin. Whether a local drop in the ATP/ADP ratio can allow local, transient desequestration and polymerization of actin either close to the plasma membrane, following platelet or neutrophil stimulation, or behind the Listeria bacterium in the host cell, while the surrounding cytoplasm contains sequestered ATP-G-actin, is an open issue raised by the present work.

Actins↗