[Prevalence of erectile dysfunction in patients consulting urological clinics: the ENJEU survey (one day national survey on prevalence of male sexual dysfunction among men consulting urologists)]. / Prévalence de la dysfonction érectile chez les patients consultant en urologie : l'enquête ENJEU (Enquête nationale de type 1 Jour sur la prévalence de la dysfonction Erectile chez des patients consultant en urologie).

OBJECTIVE: The objective of this survey was to describe medical reasons disclosed by men over 18 years old when visiting an urologist, to estimate the prevalence of male sexual dysfunction (MSD) including erectile dysfunction (ED) and to describe treatment options for ED. METHODS: This survey was performed, with Urology French Association (AFU) partnership, in 150 urological clinics, sample representative of urologists in France regarding age, geographical distribution and practice. The survey was proposed to all adults' men consulting a participating urologist the defined day (Tuesday the 19th of June 2007 or an imminent day). A total of 1848 (92.5%) patients agreed to participate; analysis was performed on 1740 patients. Information related to urological disorders, sexual dysfunctions, their treatment and their impact on the patient's life were gathered by a patient auto-questionnaire. Erectile dysfunction was assessed through the single question of John B. McKinlay. RESULTS: Among patients (mean age 63+/-14 years), 68% (IC95%=[65.2%; 70.7%]) had ED (44% severe); 25% were treated (of which 2/3 with IPDE5 alone or in association). Male sexual dysfunction was the first reason for visiting urologists (14%) following prostatic diseases (62%). About 60% of the patients had already talked about their ED to a physician, who was an urologist in 44.6% of cases. The perspective of living the rest of their life with this trouble was "unacceptable" for 21.1% of patients with ED and "fairly acceptable" for 34.4%. CONCLUSIONS: This first survey in French urologists' community emphasizes the high prevalence male sexual dysfunctions for inpatients visiting their urologists. Despite declared urologists' interest for male sexual dysfunction, the discrepancy between the high prevalence of ED and the low rate of patients consulting for this condition probably explains the low rate of patients using treatments.

Cost-effectiveness of linezolid versus vancomycin for hospitalized patients with complicated skin and soft-tissue infections in France.

UNLABELLED: Studies have shown similar clinical cure rates and shorter length of hospitalization when using linezolid compared to vancomycin in patients with complicated skin and soft-tissue infections due to suspected or proven methicillin-resistant Staphylococcus aureus (MRSA). OBJECTIVE: This study had for aim to compare the cost-effectiveness of linezolid versus vancomycin in French healthcare settings. METHOD: A decision-analytic model followed an average patient from the initiation of an empiric treatment until cure, death or second-line treatment failure. A clinical data probability was obtained from clinical trials, resource utilization data (including treatment duration and length of hospitalization) and prevalence of MRSA was obtained from a Delphi panel, and costs from published sources. RESULTS: First-line cure rate for linezolid-treated patients was 90.7% versus 85.5% for vancomycin; the total cure rates after two lines of treatment were 98.5% and 98.0%, respectively. The average total cost was 7,778euro for linezolid versus 8,777euro for vancomycin. The mean estimated length of hospitalization after two lines of treatment was 10.7 days for linezolid versus 13.3 days for vancomycin. The increased effectiveness and reduced cost lead to more frequent prescription. This did not change after one-way sensitivity analyses. CONCLUSION: Linezolid may be considered as a cost-effective treatment for patients with complicated skin and soft-tissue infections suspected to be MRSA related in France.

Types of choroidal neovascularisation in newly diagnosed exudative age-related macular degeneration.

AIM: To describe the types and location of choroidal neovascularisation (CNV) in exudative age-related macular degeneration (AMD), including vascularised pigment epithelial detatchments (PED), and most recently described subtypes, such as retinal choroidal anasmostosis, also termed "retinal angiomatous proliferation" (RAP). METHODS: Prospective multicentre consecutive descriptive case series. A total of 207 consecutive cases of newly diagnosed exudative AMD undergoing fluorescein angiography (FA) were recruited by 7 French referral hospital-based or private centres. Indocyanine green angiography (ICG) also was performed, when judged necessary by investigators. Types and location of CNV were classified by two independent experts and adjudicated by a third when discordant. RESULTS: All patients had FA, while ICG was performed in 50% of subjects. A total of 17.6% had classic CNV only, 5.4% and 8.3% had predominantly and minimally classic CNV, respectively. Occult CNV could be classified in occult CNV without PED (32.7%) and occult CNV with PED, ie, vascularised PED (23.9%). RAP was observed in 15.1% of cases, and accounted for 30% of vascularised PED. In 5.8% of the cases there was haemorrhagic AMD and 4.8% had fibrovascular scars. Lesions were mainly subfoveal (80%). Agreement between the centre's ophthalmologist and the final validated expert classification was moderate (kappa = 0.52 for location and 0.59 for type of lesion). CONCLUSION: This study confirms that newly diagnosed cases of exudative AMD are mainly occult and subfoveal. RAP appeared as a common lesion in patients with newly diagnosed exudative AMD.

[Prevalence and description of treatment with intraocular pressure-lowering topical medications in continental France]. / Prévalence et description du traitement par hypotonisants pour glaucome et hypertonie oculaire en France.

PURPOSE: The first objective of the present study was to estimate the prevalence of patients treated for glaucoma or ocular hypertension (OHT) with an IOP (intraocular pressure) -lowering topical medication. Subjects were 40 years old and living in continental France. The second objective was to describe the type of therapy, compliance, and the quality of life of the subjects treated. METHODS: This study was a telephone survey conducted in a general population on a representative sample of 5726 subjects in 2004. Subjects treated for glaucoma or OHT were defined as those declaring use of eye drops for more than 1 month and mentioning one of the 52 registered IOP-lowering topical medications and/or stating that they were taking eye drops for glaucoma or OHT. Quality of life was measured using the GlauQOL-17 questionnaire. RESULTS: 237 subjects (4.1%) corresponded to this definition. The prevalence increased with age, from 0.7% in subjects aged 40-44 years to 10.6% in those aged 80 years or more. The age-standardized prevalence was higher in the Paris area (5.3%), the Mediterranean (4.5%), and the Southwest (4.7%), and lower in rural areas (3.2%), and in subjects with less than high school education (2.9%). Beta-blockers (49.5%) and prostaglandins (37.4%) were the most frequent treatments, followed by carbonic anhydrase inhibitors (15.3%), sympathomimetics (5.7%), and myotics (1.3%). Compliance was not total in 39% of the patients (22% of those treated with once-a-day dosage). DISCUSSION: The results of the present study are in line with French health authority data and with the results of the French "Glaucoma and intraocular high pressure, one day" study results. Moreover, the lower prevalence in regions with low medical density, low socioeconomic level, and rural areas, suggest that glaucoma or OHT might be underdiagnosed. CONCLUSION: On the basis of this first nationwide study, it can be estimated that the prevalence of subjects treated with IOP-lowering topical medications is about 1.2 million in France. The number of daily doses appears to influence compliance.

[Clinical tolerance of antiglaucoma eyedrops with and without a preservative. Results of an unpublished survey in Europe]. / Tolérance clinique des collyres antiglaucomateux conservés et non conservés. Résultats d'une enquête inédite en Europe.

PURPOSE: To evaluate the clinical tolerance of antiglaucoma eye drops with and without a preservative in routine practice. METHODS: A total of 125 ophthalmologists in private practice located throughout France examined altogether 919 glaucomatous patients treated with eye drops containing a preservative or not. For some patients the examination was repeated during a second visit. Functional signs as well as conjunctival and corneal examination results were recorded. RESULTS: The proportion of patients experiencing discomfort or pain during instillation was 58% for eye drops containing a preservative and 30% for eye drops with no preservative (p < 0.001). Moreover, the proportion of patients presenting at least one symptom of eye irritation (sensation of itching or burning, sensation of a foreign body in the eye, and flow of tears) was greater with preservative-containing eye drops (53% vs 34%; p < 0.001). The experience of discomfort during instillation was more often associated with problems later on. The patient's complaints were correlated with objective signs of conjunctival (conjunctival redness, conjunctival follicles), or corneal (superficial punctuate keratitis) damages. A higher proportion of patients treated with eye drops containing a preservative showed at least one conjunctival sign (52% vs 35%; p = 0.001) or superficial punctuate keratitis (12% vs 4%; p = 0.01). In 164 patients, whose treatment was changed from eye drops containing a preservative to eye drops with no preservative and who were examined a second time (mean interval between visits: 3.3 months) the frequency of all symptoms and objective signs fell by a factor of 3 to 4 (p < 0.001). CONCLUSION: Patient complaints and objective damage to the ocular surface are more frequent among patients treated with eye drops containing a preservative. Ocular surface damage due to preservatives is partially reversible. Patient complaints should consequently be taken into account and preservative-free eye drops should be prescribed when ever possible.

Transcription factors and aldolase B gene expression in microdissected renal proximal tubules and derived cell lines.

Renal expression of the aldolase B isoenzyme and transcription factors previously shown to regulate the aldolase B gene promoter in the liver were analyzed in whole kidney, microdissected tubules, and the two PKSV-PCT and PKSV-PR proximal tubule cell lines derived from transgenic mice. Aldolase B gene expression appeared restricted to the proximal tubule, the site where HNF1 alpha, HNF1 beta, C/EBP alpha, and DBP transcripts were also abundant. Compared to the liver, another organ synthesizing aldolase B, proximal tubules from the kidney were characterized by the absence of HNF3 and the presence of higher ratio of HNF1 beta/HNF1 alpha transcripts. The same features were conserved in both PKSV-PCT and PKSV-PR proximal tubule cell lines. Transactivation experiments in PKSV-PCT cultured cells showed that HNF1 alpha, C/EBP alpha, and DBP behave as transactivators of the 190-bp aldolase B gene promoter, and that HNF1 beta had a low transactivating efficiency. HNF1 beta, as well as HNF3, antagonized the HNF1 alpha-dependent transactivation of the aldolase B promoter. The fact that both HNF1 beta and HNF3 factors play similar negative roles by competitively binding close to or on the HNF1 site could suggest that, in proximal tubule renal cells, HNF1 beta has the same attenuator effect on the aldolase B gene promoter as HNF3 in hepatocytes. Thus, these results indicate that such models of established renal tubule cell lines, which have conserved the same features of parental cells, represent valuable tools for studies of the regulation of genes expressed in proximal tubules of the kidney.

Influence of the content in transcription factors on the phenotype of mouse hepatocyte-like cell lines (mhAT).

We have described new well-differentiated mouse hepatocyte-like cell lines (mhAT) derived from transgenic mice expressing simian virus 40 large T antigen under the control of antithrombin III gene promoter (Exp. Cell Res. (1992) 200, 175-185). In an attempt to understand the phenotypic variations of the different cell lines, we analyzed their content in liver-specific transcription factors at the level of both the proteins, by gel shift analysis, and the mRNA, by quantitative reverse-transcription PCR. Moreover, the transactivating ability of endogenous HNF1 alpha and C/EBP alpha was also evaluated by measuring the activity of transfected synthetic promoters consisting of DNA element homopolymers upstream of a TATA box. High levels of HNF1, HNF3, and HNF4 transcription factors were maintained in mhAT cells. In contrast, C/EBP alpha was much more variable between the different cell lines and was less abundant than it was in vivo, in the liver. We investigated the influence of HNF1 alpha and C/EBP alpha on the activity of transfected liver-specific promoters. HNF1 alpha was not limiting for the activity of transfected liver-type pyruvate kinase and albumin promoters. In contrast, the activity of the albumin promoter in the different lines was clearly dependent on the C/EBP alpha content, which seems, therefore, to be an essential factor modulating the expression of this gene in HNF1 alpha-containing cells. This work shows that the correlations between promoter activities and transacting factor contents in a panel of well-differentiated cultured cells can be used to determine the respective role of transcription factors on the strength of some promoters.

Diet-dependent carcinogenesis of pancreatic islets and liver in transgenic mice expressing oncogenes under the control of the L-type pyruvate kinase gene promoter.

The liver-type pyruvate kinase (L-PK) gene is controlled positively by insulin and carbohydrates, negatively by glucagon and fasting. Diet-inducible models of carcinogenesis were obtained using the L-PK gene promoter and regulatory sequences to control the expression of c-myc and SV40 T oncogenes in transgenic mice. L-PK/c-myc and L-PK/Tag animals fed a carbohydrate-rich diet developed hepatocarcinomas. In addition, L-PK/Tag animals developed diet-dependent, aggressive endocrine pancreatic tumors, preceded by islet hyperplasia involving the different analysed cell populations (alpha, beta and delta). Expression of the L-PK gene was demonstrated in pancreatic tumors, in rat isolated islets and in rat insulinoma-derived cells (RIN line), revealing a new tissue specificity of the L-PK gene. Our results suggest that this gene may be expressed in islet progenitor cells from which the different mature endocrine cells derive.

Gene expression in hepatocyte-like lines established by targeted carcinogenesis in transgenic mice.

New hepatocyte-like cell lines (mhAT) were derived from the liver of a transgenic mouse expressing SV40 early genes under the direction of the liver-specific antithrombin III gene promoter (ATIII-TSV40). Their differentiated phenotypes were improved and stabilized by the use of liver-specific growth media (arginine-free, glucose-free, or low-fructose/glucose-free medium). The best differentiated lines display a very high level of albumin, transferrin, and L-type pyruvate kinase (L-PK) gene expression that is comparable to that observed in the mouse liver. Abundance of the aldolase B and phosphoenolpyruvate carboxykinase (PEPCK) transcripts varied from 5 to 35% of the in vivo concentrations while abundance of the alpha-fetoprotein and phenylalanine hydroxylase transcripts remained very low. Hormonal (cAMP and insulin) and nutritional (glucose) gene controls of PEPCK and L-PK were, at least partially, conserved. mhAT cells are readily transfectable by the calcium phosphate coprecipitation technique and exhibit a liver-specific pattern of expression of exogenous genes. Thus, mhAT cells seem suitable for the analysis of the regulatory regions involved in the tissue-specific transcription of genes. This work demonstrates, therefore, the great efficiency of targeted carcinogenesis in transgenic mice to create new differentiated cell lines. The availability of various lines of liver-specific cells with different phenotypes will constitute useful tools to establish correlations between expression of trans-acting factors and control of the phenotype.

Proteins binding to the liver-specific pyruvate kinase gene promoter. A unique combination of known factors.

A 183 base-pair fragment of the liver-specific promoter of the L-type puruvate kinase (L-PK) gene has been shown by transfection assay to be sufficient to confer a tissue-specific expression to a reporter gene. The proteins binding in vitro to this fragment have been investigated by a combination of DNase I footprinting, gel retardation of synthetic oligonucleotides and ultraviolet cross-linking. Four proteins from liver nuclear extracts bind to the investigated fragment. They were called, from 3' to 5', L1 to L4 binding factors. The L1 site (nucleotides -95 to -66 with respect to the cap site) binds hepatocyte nuclear factor 1 (HNF1), a liver-specific protein. The L2 site (nucleotides -114 to -97) binds the ubiquitous nuclear factor 1 (NF1), or a related factor. The L3 site (nucleotides -144 to -126) binds liver factor A1 (LF-A1), another liver-specific protein. Finally, the L4 site (nucleotides -168 to -145) binds major late transcription factor (MLTF/USF/UEF), an ubiquitous protein. Each of these proteins has been detected in other liver-specific promoters, but their combination is unique to the liver-specific promoter of the L-PK gene.

Transfection of hepatic genes into adult rat hepatocytes in primary culture and their tissue-specific expression.

We describe in this paper a method for studying transient gene expression in a primary culture of adult rat hepatocytes. After isolation by collagenase perfusion, hepatocytes in a monolayer were transfected with foreign DNA by the calcium phosphate precipitation technique during the first 24 hours after plating. When they were transfected with a plasmid containing the gene for chloramphenicol acetyltransferase driven by the early promoter of simian virus 40, hepatocytes reproducibly expressed high levels of chloramphenicol acetyltransferase (CAT); this transient expression was much higher than that obtained with the rat hepatoma cell line H4II. Different medium conditions have been tested; an optimal level of CAT activity can be obtained using a serum-free, hormonally defined medium. Using these techniques, we have investigated the expression of liver-specific genes transferred into hepatocytes. We show that the L-pyruvate kinase promoter is active in these hepatocytes while it is silent in fibroblasts. Moreover, the use of serum-free medium may allow investigation of the role of hormones and nutrients in cells which respond normally to these effectors.