[Pre- and post-prostatectomy rehabilitation by the certified nurse-urologist pair (REHAB): Feasibility study]. / Parcours de réhabilitation par le binôme IDE-urologue avant et après prostatectomie (REHAB) : étude de faisabilité.

INTRODUCTION: The functional results of radical prostatectomy are a crucial issue for patients to resume fulfilling sexuality. We assessed the feasibility of a care pathway dedicated to sexual rehabilitation in order to improve information, screening of risk situations and the implementation of therapeutic measures. METHODS: From January to May 2023, sexually active patients under 75 years of age undergoing prostatectomy for cancer were offered the opportunity to participate in two sexual rehabilitation sessions (REHAB) led by a nurse-urologist pair. The sessions took place in parallel with the care pathway already in place before and after surgery. The evaluations were carried out by carrying out questionnaires and a clinical examination. A satisfaction questionnaire was given to the patient after the two sessions to assess the format and relevance of the sessions. RESULTS: Fifteen patients were included in the REHAB program. All patients attended both sessions in person and the majority of them (91%) felt they had obtained new information for their rehabilitation. Post-operatively at 6 weeks, 82% of patients were dissatisfied with their sexuality (compared to 64% pre-operatively), Five patients (33%) had regained orgasmic abilities and 20% (n=3) had a penetrative ability. The average IIEF5 score was 19 (6-28) compared to 22.5 pre-operatively (14-30). All patients would recommend these sessions. CONCLUSION: The REHAB sexual rehabilitation program after prostatectomy could be implemented with significant patient adherence and satisfaction.

[Endoscopic papillary abnormalities and urinary stone recognition (EPSR): How and why?] / Reconnaissance endoscopique des anomalies papillaires et des calculs urinaires (REPC) : comment et quel intérêt ?

INTRODUCTION: The purpose of this article is to present the endoscopic papillary abnormalities and stone recognition (EPSR) to state-certified nurses (IDE and IBODE) working in the operating room. METHODS: This article is based on a literature review and the author's experience concerning the endoscopic papillary abnormalities and stone recognition. RESULTS: Since the advent of minimally invasive surgery and the laser, stones are no longer sent as one piece to laboratories, but fragmented. This has made it more difficult for biologists to fully analyze the stones, because they have less morphological data than before. Therefore, endoscopic papillary abnormalities and stone recognition have positioned themselves as tools that can compensate for this loss of information. They play a pivotal role in the identification of the lithogenesis cause, and thus allow a recurrence risk reduction of stones. CONCLUSION: Endoscopic papillary abnormalities and stone recognition are recent tools that require learning. However, the benefit of their uses is proven and is necessary for a complete management of urolithiasis.

[Double J stent removal's cost estimate using a re-sterilizable fibroscope in a French private structure]. / Évaluation du coût en structure privée d'une ablation de sonde double J en externe à l'aide d'un fibroscope re-stérilisable.

In front of the arrival of new devices intended to simplify the removal of double J stent, it poses the problem of the knowledge of the real cost of such an ablation under the current conditions of realization. MATERIAL AND METHOD: This is a monocentric economic evaluation of cost and remuneration needed data-gathering of quotation (CCAM, GHS/SE, …), estimate of the associated costs of wear and damping of the endoscopic equipments (endoscopes, cables, …), estimate of the cost of sterilization, estimate of the associated costs to the intervention of staff (Auxiliary nurse [AS] and Nurse [IDE]) with timing of the various tasks. RESULTS: Quotation CCAM JCGE004 (48€) gives access to fixed price SE1 (73.71€ for private clinic, and 75.89€ for public institution) without hospitalization nor anaesthesia. The costs were reported to an act of single double J removal. Concerning the equipments: 4.42€HT for the fibroscopes, graspers, cable and light. The costs of sterilization were: 17.95€HT. The timed workforce's costs were: 7.61-9.51€ for AS and 9.92-10.84€ for IDE. The cost of consumable was about 1.37 €HT, by excluding the common base from the extractions (1.876€HT). The total costs in France in 2016 were thus about 47.4 to 50.496€ including all taxes. CONCLUSIONS: This estimate will be used certainly for reflection on the investments and the future studies of the economic impact of the new devices of extraction, by correlating it of course with the various maintenance contracts from each institution. LEVEL OF PROOF: 4.

Role of a new mammalian gene family in the biosynthesis of very long chain fatty acids and sphingolipids.

Whereas the physiological significance of microsomal fatty acid elongation is generally appreciated, its molecular nature is poorly understood. Here, we describe tissue-specific regulation of a novel mouse gene family encoding components implicated in the synthesis of very long chain fatty acids. The Ssc1 gene appears to be ubiquitously expressed, whereas Ssc2 and Cig30 show a restricted expression pattern. Their translation products are all integral membrane proteins with five putative transmembrane domains. By complementing the homologous yeast mutants, we found that Ssc1 could rescue normal sphingolipid synthesis in the sur4/elo3 mutant lacking the ability to synthesize cerotic acid (C(26:0)). Similarly, Cig30 reverted the phenotype of the fen1/elo2 mutant that has reduced levels of fatty acids in the C(20)-C(24) range. Further, we show that Ssc1 mRNA levels were markedly decreased in the brains of myelin-deficient mouse mutants known to have very low fatty acid chain elongation activity. Conversely, the dramatic induction of Cig30 expression during brown fat recruitment coincided with elevated elongation activity. Our results strongly implicate this new mammalian gene family in tissue-specific synthesis of very long chain fatty acids and sphingolipids.

The immunosuppressant SR 31747 blocks cell proliferation by inhibiting a steroid isomerase in Saccharomyces cerevisiae.

SR 31747 is a novel immunosuppressant agent that arrests cell proliferation in the yeast Saccharomyces cerevisiae, SR 31747-treated cells accumulate the same aberrant sterols as those found in a mutant impaired in delta 8- delta 7-sterol isomerase. Sterol isomerase activity is also inhibited by SR 31747 in in vitro assays. Overexpression of the sterol isomerase-encoding gene, ERG2, confers enhanced SR resistance. Cells growing anaerobically on ergosterol-containing medium are not sensitive to SR. Disruption of the sterol isomerase-encoding gene is lethal in cells growing in the absence of exogenous ergosterol, except in SR-resistant mutants lacking either the SUR4 or the FEN1 gene product. The results suggest that sterol isomerase is the target of SR 31747 and that both the SUR4 and FEN1 gene products are required to mediate the proliferation arrest induced by ergosterol depletion.

Compliance with clothing regulations and traffic flow in the operating room: a multi-centre study of staff discipline during surgical procedures.

This multi-centre study assessed operating room (OR) staff compliance with clothing regulations and traffic flow during surgical procedures. Of 1615 surgical attires audited, 56% respected the eight clothing measures. Lack of compliance was mainly due to inappropriate wearing of jewellery (26%) and head coverage (25%). In 212 procedures observed, a median of five people [interquartile range (IQR) 4-6] were present at the time of incision. The median frequency of entries to/exits from the OR was 10.6/h (IQR 6-29) (range 0-93). Reasons for entries to/exits from the OR were mainly to obtain materials required in the OR (N=364, 44.5%). ORs with low compliance with clothing regulations tended to have higher traffic flows, although the difference was not significant (P=0.12).

Human lamin B receptor exhibits sterol C14-reductase activity in Saccharomyces cerevisiae.

Lamin B receptor (LBR), a nuclear protein of avian and mammalian cells, contains an hydrophobic domain that shares extensive structural similarities with the members of the sterol reductase family. To test if the sterol-reductase-like domain of LBR could be enzymatically competent, several sterol reductase-defective strains of Saccharomyces cerevisiae were transformed with a human-LBR expressing vector. LBR production did not change the ergosterol biosynthesis defect in an erg4 mutant impaired in sterol C24(28) reductase. In contrast, the sterol C14 reduction step and ergosterol prototrophy were restored in LBR-producing erg24 transformants which lack endogenous sterol C14 reductase. To test the effects of C14 reductase inhibitors on LBR activity, we constructed EMY54, an ergosterol-requiring strain that is devoid of both sterol C8-C7 isomerase and sterol C14 reductase activities. EMY54 cells recovered the capability of synthesizing ergost-8-en-3beta-ol upon transformation with a vector that expressed either yeast sterol C14 reductase or hLBR. In addition, growth in sterol-free medium was restored in these transformants. Sterol biosynthesis and proliferation of LBR-producing cells were found to be highly susceptible to fenpropimorph and tridemorph, but only moderately susceptible to SR 31747. Our results strongly suggest that hLBR is a sterol C14 reductase.

[Ureterolysis: technique, indications]. / Urétérolyse: technique, indications.

Ureteral obstruction due to idiopathic retroperitoneal fibrosis is a rare but severe clinical problem. The open approaches, as well as surgical techniques used to prevent stenosis recurrence, are described. Ureterolysis remains the procedure to relieve ureteral obstruction. The ureter is dissected and freed from the fibrotic process, and then separated to prevent the recurrence of the stenosis. Recently, the development of Laparoscopic urology has allowed for minimal invasive treatment of many urological problems. We present our technique of ureterolysis for extrinsic ureteral obstruction. Advantages and complications of each method are considered and indications are proposed.

Expression of a cloned Saccharomyces cerevisiae gene (URA1) is controlled by a bacterial promoter in E. coli and by a yeast promoter in S. cerevisiae.

The expression of a cloned yeast URA1 gene in Escherichia coli and in Saccharomyces cerevisiae was studied. In E. coli, only one orientation of the cloned yeast DNA segment inserted into the bacterial vector (pBR322) allows URA1 expression. Moreover, the permissive orientation changes with the cloning site. The absence of URA1 expression in E. coli can be corrected by the spontaneous integration into the cloned yeast DNA of a 0.9-kb bacterial DNA. Several copies of such a bacterial IS element have been detected in the host E. coli genome. The results strongly suggest that, in E. coli, transcription of the yeast URA1 needs a prokaryotic promoter for its initiation. In S. cerevisiae, the expression of non-chromosomally cloned URA1 does not depend on the orientation of the cloned fragment. Furthermore, it remains under the control of a nuclear regulatory gene (pprX-1) which constitutively enhances the expression of URA1 as well as URA3 at the transcriptional level. Therefore, in S. cerevisiae, transcription of non-chromosomally cloned URA1 involves a physiological yeast promoter cloned along with the structural part of the gene.

High-level production of a peroxisomal enzyme: Aspergillus flavus uricase accumulates intracellularly and is active in Saccharomyces cerevisiae.

Strains of Saccharomyces cerevisiae producing Aspergillus flavus uricase (Uox) have been constructed. An artificial promoter which combined the upstream and downstream sequences of the GAL7 and ADH2 promoters, respectively, was found to be efficient in directing the synthesis of uaZ mRNAs encoding Uox. A good proportionality between the copy number of the uaZ expression cassette and the level of Uox production was found in the range of 1-10 copies. Transformants accumulated active and soluble Uox to a level exceeding 13% of total protein, as deduced from enzymatic assays. This relative level could be improved two- to threefold by using a recipient strain in which the wild-type GAL4 gene had been deleted and which expressed a GAL4 construct placed under the control of the ADH2 promoter.

High-level production of a human membrane protein in yeast: the peripheral-type benzodiazepine receptor.

The human peripheral-type benzodiazepine receptor (PBR) has been produced in Saccharomyces cerevisiae where it retains its pharmacological properties [Riond et al., Eur. J. Pharmacol. 208 (1991) 307-312]. As the rate of production was low, we analysed the mRNA level, the effect of variation of the 5' sequence and the production in mitochondria. Translation rather than transcription or targeting was found to be the main limiting factor. We were able to produce a chimeric PBR, with an N-terminal extension, to a very high level in the yeast mitochondrial membrane.

Sterol metabolism and ERG2 gene regulation in the yeast Saccharomyces cerevisiae.

Certain exogenously-supplied sterols, like ergost-8-enol, are efficiently converted into ergosterol in yeast. We have taken advantage of this property to study the regulation of the Delta8-Delta7-sterol isomerase-encoding ERG2 gene in an ergosterol auxotrophic mutant devoid of squalene-synthase activity. Ergosterol starvation leads to an 8-16-fold increase in ERG2 gene expression. Such an increase was also observed in wild-type cells either grown anaerobically or treated with SR31747A a sterol isomerase inhibitor. Exogenously-supplied zymosterol is entirely transformed into ergosterol, which represses ERG2 transcription. By contrast, exogenously-supplied ergosterol has little or no effect on ERG2 transcription.

Molecular basis for the different binding properties of benzodiazepines to human and bovine peripheral-type benzodiazepine receptors.

The 18 kDa peripheral benzodiazepine receptor (PBR) can be labelled by benzodiazepines, such as Ro5-4864, and isoquinoline carboxamides such as PK11195. These two compounds are reversible competitive inhibitors of each other. However, while the binding affinity of Ro5-4864 varies enormously across species, PK11195 always displays high affinity, suggesting that their binding domains are overlapping but not identical. We report here that recombinant human and bovine PBR produced in yeast, a microorganism devoid of endogenous PBR, can be labelled with [3H]PK11195, but only the human receptor can be labelled with [3H]Ro5-4864. Furthermore, we identified, through the binding analysis of human-bovine chimaeric receptors, a region near the C-terminal end of the PBR, with only five non-conserved amino acids between human and bovine sequences, as responsible for the difference in high affinity binding of Ro5-4864 to the two receptors.

Expression and pharmacological characterization of the human peripheral-type benzodiazepine receptor in yeast.

Recently we cloned the cDNA coding for the putative human peripheral-type benzodiazepine receptor (hPBR). This report describes the expression of this cDNA in Saccharomyces cerevisiae and the characterization of the recombinant protein. The expression was achieved by placing the receptor cDNA under the control of a galactose-regulated artificial promoter. After galactose induction, the transformed cells expressed a functional hPBR which displayed a Kd for the specific peripheral-type ligand [3H]PK11195 of 9.9 +/- 1.3 nM and a maximal binding capacity of 249,300 +/- 50,400 sites/cell. The pharmacological characterization of the recombinant receptor, determined in competitive ligand binding experiments, agrees closely with that described for the natural receptor expressed by human cells. Furthermore, the binding was stereospecific as shown by the displacement of the [3H]PK11195 binding by PK14067 (-Q1) and not by PK14068 (+Q1). Photolabeling experiments showed that transformed cells expressed a 18 kDa protein which was specifically labeled with [3H]PK14105. Altogether these results show that the cDNA transfected in yeast encodes a 18 kDa protein with the expected characteristics of the hPBR.

Genetic improvements of an industrial strain of Aspergillus flavus for urate oxidase production.

Urate oxidase, an enzyme used in human therapy, is currently produced industrially by a strain of Aspergillus flavus. Two strategies of strain improvement were tested in order to obtain higher yields of urate oxidase. The first one, based on a classical mutation-selection protocol, led to the isolation of a mutant strain that overproduced uricase two-fold as compared to the industrial strain. The second one consisted in the construction of transformed strains that had integrated multiple copies of a urate oxidase-expression vector. A twenty-fold improvement in urate oxidase was obtained by this method.

Self-cloning in filamentous fungi: application to the construction of endothiapepsin overproducers in Cryphonectria parasitica.

The filamentous ascomycete fungus Cryphonectria parasitica naturally secretes endothiapepsin, an aspartic proteinase. It is cultured on a commercial scale as a source of the milk-clotting enzyme for cheese making. Our objective was to increase enzyme production of an industrial C. parasitica strain by a new technique of self-cloning; it consisted in the screening for transformants producing higher levels of endothiapepsin and having integrated only the DNA fragment of interest. Such genetically improved strains that are devoid of any foreign genes should be more readily acceptable for the production of food-grade enzymes.

Constitutive mutants for orotidine 5 phosphate decarboxylase and dihydroorotic acid dehydrogenase in Saccharomyces cerevisiae.

A mutant yeast constitutive for synthesis of both dihydroorotic acid dehydrogenase and orotidine 5' phosphate decarboxylase has been isolated. This phenotype is due to a single gene unlinked to any of the five genes of the pyrimidine pathway. This gene, called ppr1, induces the unlinked genes ural and ura3. Non-chromosomal cloned ura3 is also under the control of ppr1.