[Integration of a hospital pharmacist in the ICU]. / Intégration d'un pharmacien hospitalier en service de réanimation.

The French regulatory system strongly encourages strict regulation of health products' production and distribution, especially concerning risk management and economic aspects. An ICU is an unusual environment for a local pharmacy practice (a nurse for every 2.5 patients, continuous adaptation of therapeutics…). However, a literature review reports interesting data concerning risk management and economics. This article aims to relate the experience of a pharmacist integration in a French teaching hospital ICU (half-time position).

[Innovative therapeutic strategies for intravesical drug administration]. / Stratégies thérapeutiques innovantes pour l'administration médicamenteuse intravésicale.

INTRODUCTION: Perspectives for innovative pharmaceutical molecules and intravesical administration of pharmacological agents are presented in the present review carried out from a recent literature. MATERIALS AND METHODS: This review of the literature was built by using the PubMed and ScienceDirect databases running 20keywords revealing 34publications between 1983 and 2012. The number of referenced articles on ScienceDirect has increased in recent years, highlighting the interest of scientists for intravesical drug administration and the relevance of innovating drug delivery systems. RESULTS: Different modalities of intravesical administration using physical (e.g., iontophoresis, electroporation) or chemical techniques (e.g., enzyme, solvent, nanoparticles, liposomes, hydrogels) based on novel formulation methods are reported. Finally, the development of biopharmaceuticals (e.g., bacillus Calmette-Guérin, interferon α) and gene therapies is also presented and analyzed in this review. CONCLUSION: The present review exhibits new development in the pipeline for emerging intravesical drug administration strategies. Knowledge of all these therapies allows practitioners to propose a specific and tailored treatment to each patient with limiting systemic side effects.

Novel interaction partners of Bardet-Biedl syndrome proteins.

Bardet-Biedl syndrome (BBS) is a rare, developmental disorder characterized by six major symptoms: rod-cone dystrophy, obesity, polydactyly, renal abnormalities, learning difficulties, and hypogonadism. Secondary features include cardiac and hepatic anomalies, metabolic disturbancies, and hearing loss. BBS is genetically heterogeneous with 12 disease genes (BBS1-BBS12) described thus far. Current data suggest a functional disturbance in ciliary function and intraflagellar transport being associated with the phenotype. However, the precise functions of the BBS proteins have yet to be elucidated. This study focuses on the detection of protein factors interacting with BBS proteins. Applying yeast two-hybrid (Y2H) technology we found a series of novel, functionally potentially plausible binding partners of BBS1, BBS2, BBS4, and BBS7. Protein interactions were supported by coimmunoprecipitation analyses (ALDOB, EPAS1) and substantiated by colocalization studies at the subcellular level (ALDOB, EXOC7, FLOT1, KRT18, PAX2). Our work provides new insights into the understanding of BBS interactions and thus their biological function.

Divergence between the high rate of p53 mutations in skin carcinomas and the low prevalence of anti-p53 antibodies.

Circulating anti-p53 antibodies have been described and used as tumoural markers in patients with various cancers and strongly correlate with the p53 mutated status of the tumours. No study has yet looked at the prevalence of such antibodies in skin carcinoma patients although these tumours have been shown to be frequently p53 mutated. Most skin carcinoma can be diagnosed by examination or biopsy, but aggressive, recurrent and/or non-surgical cases' follow up would be helped by a biological marker of residual disease. We performed a prospective study looking at the prevalence of anti-p53 antibodies using an ELISA technique in a series of 105 skin carcinoma patients in comparison with a sex- and age-matched control skin carcinoma-free group (n = 130). Additionally, p53 accumulation was studied by immunohistochemistry to confirm p53 protein altered expression in a sample of tumours. Anti-p53 antibodies were detected in 2.9% of the cases, with a higher prevalence in patients suffering from the more aggressive squamous cell type (SCC) of skin carcinoma (8%) than for the more common and slowly growing basal cell carcinoma type or BCC (1.5%). p53 protein stabilization could be confirmed in 80% of tumours studied by IHC. This low level of anti-p53 antibody detection contrasts with the high rate of p53 mutations reported in these tumours. This observation shows that the anti-p53 humoral response is a complex and tissue-specific mechanism.

Searching for excystment-regulated genes in Sterkiella histriomuscorum (Ciliophora, Oxytrichidae): a mRNA differential display analysis of gene expression in excysting cells.

In the absence of food, the oxytrichid Sterkiella histriomuscorum transforms like many ciliates into resting cysts. When transferred back into feeding medium, the cyst re-transforms into a vegetative cell. The entry into and exit from the dormant cyst stage are complex developmental processes still poorly investigated at the molecular level. Assuming that these changes in state could involve changes in gene expression, we have used the technique of mRNA differential display to detect differentially expressed genes in cysts and two different stages of excysting cell. Variation in the temporal expression pattern of transcripts could be detected and, in using an inverse-PCR strategy on circularized macronuclear DNA, we have sequenced the macronuclear genes of three of the isolated cDNAs. which correspond to 1) a nucleotide-binding domain-encoding gene, 2) a DHHC-domain-carrying gene, and 3) a phosphatase type 2C-encoding gene. For the first two genes, Northern blot analyses supported an excystment-associated regulated gene expression. We discuss their possible role during excystment and we show that the combination of differential display and inverse PCR constitutes a powerful approach to isolate excystment-regulated genes in hypotrichs.

Sensitivity of a clinical scale of balance and gait in frail nursing home residents.

PURPOSE: The purpose of this paper was to describe this application, and to assess the sensitivity of the application when evaluating clinical interventions for improving balance and gait. METHODS: The records of fifteen consecutive patients referred to physical therapy for mobility problems or recent falls were reviewed for this study. The subjects were evaluated upon initial referral to physical therapy and after 30 days of treatment. Treatment was developed around the problems noted on initial evaluation and applied 5 days/week for 20 sessions. RESULTS: A Wilcoxon signed-rank comparison of the initial and 30 day re-evaluation of the balance and the gait assessment indicated that significant improvement had occurred in the balance scores (Z = -3.20, p = 0.001) and the gait scores (Z = -2.82, p = 0.005) in this group. CONCLUSION: These assessments are sensitive to clinical improvements in mobility among frail elders.

Transcription of the nfrA-ywcH operon from Bacillus subtilis is specifically induced in response to heat.

The NfrA protein, an oxidoreductase from the soil bacterium Bacillus subtilis, is synthesized during the stationary phase and in response to heat. Analysis of promoter mutants revealed that the nfrA gene belongs to the class III heat shock genes in B. subtilis. An approximate 10-fold induction at both the transcriptional and the translational levels was found after thermal upshock. This induction resulted from enhanced synthesis of mRNA. Genetic and Northern blot analyses revealed that nfrA and the gene downstream of nfrA are transcribed as a bicistronic transcriptional unit. The unstable full-length transcript is processed into two short transcripts encoding nfrA and ywcH. The nfrA-ywcH operon is not induced by salt stress or by ethanol. According to previously published data, the transcription of class III genes in general is activated in response to the addition of these stressors. However, this conclusion is based on experiments which lacked a valid control. Therefore, it seems possible that the transcription of all class III genes is specifically induced by heat shock.

P16 UV mutations in human skin epithelial tumors.

The p16 gene expresses two alternative transcripts (p16alpha and p16beta) involved in tumor suppression via the retinoblastoma (Rb) or p53 pathways. Disruption of these pathways can occur through inactivation of p16 or p53, or activating mutations of cyclin dependant kinase 4 gene (Cdk4). We searched for p16, Cdk4 and p53 gene mutations in 20 squamous cell carcinomas (SSCs), 1 actinic keratosis (AK), and 28 basal cell carcinomas (BCCs), using PCR-SSCP. A deletion and methylation analysis of p16 was also performed. Six different mutations (12%) were detected in exon 2 of p16 (common to p16alpha and p16beta), in five out of 21 squamous lesions (24%) (one AK and four SCCs) and one out of 28 BCCs (3.5%). These included four (66%) ultraviolet (UV)-type mutations (two tandems CC : GG to TT : AA transitions and two C : G to T : A transitions at dipyrimidic site) and two transversions. P53 mutations were present in 18 samples (37%), mostly of UV type. Of these, only two (one BCC and one AK) harboured simultaneously mutations of p16, but with no consequence on p16beta transcript. Our data demonstrate for the first time the presence of p16 UV induced mutations in non melanoma skin cancer, particularly in the most aggressive SCC type, and support that p16 and p53 are involved in two independent pathways in skin carcinogenesis.

The human pH aldolase A promoter directs widespread but muscle-predominant expression in transgenic mice.

In order to identify regulatory elements that direct widespread in vivo expression of a linked gene, we have examined one of the human aldolase A alternative promoters, the ubiquitous pH promoter, which is active in most foetal and adult tissues. We have used the pH promoter region to drive expression of an heterologous CAT reporter gene in transgenic mice. We show that a short 820 bp pH promoter fragment is able to confer a ubiquitous and reproducible activity pattern on the CAT reporter gene in most of the transgenic lines analysed, with a particularly high level of expression in adult skeletal muscle. Activity of this transgene was detected from early embryonic stages. Therefore, this pH promoter region appears to be a powerful tool to direct ubiquitous and early expression of a transgene in vivo. Deletion analysis revealed that: (i) the region between -651 and -369 bp relative to the pH promoter transcription start site includes DNA elements capable of overriding effects of the surrounding chromatin at the integration site, (ii) the region between -285 and -211 bp is involved in pH promoter tissue-specific expression pattern in skeletal muscle and/or nervous tissues, (iii) the region located between -211 and -108 bp is necessary for its ubiquitous and muscle-predominant activity and (iv) the most proximal region downstream from -108 bp is still sufficient to confer an activity in brain and lung.

The sigmaD-dependent transcription of the ywcG gene from Bacillus subtilis is dependent on an excess of glucose and glutamate.

We investigated the function and transcriptional regulation of ywcG. The protein is essential for Bacillus subtilis. Biochemical characterization of the protein revealed that it is an FMN-containing NADPH oxidase. ywcG is transcribed throughout the whole life cycle of B. subtilis. The start point of transcription is preceded by potential promoter sequences for sigmaA, sigmaB and sigmaD. A boost in transcription occurs at the beginning of stationary phase in complex media containing glutamate and glucose. The induction of transcription at the beginning of stationary phase needs the activity of a different alternative sigma-factor sigmaD. ywcG is, therefore, the first gene with a putative role in energy metabolism from B. subtilis that is transcribed in a sigmaD-dependent fashion, but its regulation is unique and the reverse of that described for all other sigmaD-dependent genes.

Independence and interdependence of the three human aldolase A promoters in transgenic mice.

The human aldolase A gene is transcribed from three alternative promoters, clustered in a small 1.6-kb DNA domain. In transgenic mice, the upstream pN and the downstream pH promoters are ubiquitous, whereas the pM promoter, located between pN and pH, is activated specifically in fast skeletal muscles. A strong ubiquitous enhancer, lying upstream of the pH promoter, is necessary for both pN and pH ubiquitous activities, whereas a fast-muscle-specific enhancer, located upstream of the pM promoter, is required for pM-specific activation. In the present study, we use the transgenic mice model to further investigate the contribution of these two regulatory elements to the overall control of these three promoters. We confirm that the pM and pH promoters are activated independently of each other and, in particular, we show that the activation of pM in fast muscle is not responsible for the downregulation of the downstream pH in this tissue. By contrast, the pN promoter needs the presence of both enhancers to reproduce its correct pattern of activity and is unable to function autonomously in vivo.

Fast-muscle-specific expression of human aldolase A transgenes.

The expression of the human aldolase A gene is controlled by three alternative promoters. In transgenic mice, pN and pH are active in all tissues whereas pM is activated specifically in adult muscles composed mainly of fast, glycolytic fibers. To detect potential regulatory regions involved in the fast-muscle-specific activation of pM, we analyzed DNase I hypersensitivity in a 4.3-kbp fragment from the 5' end of the human aldolase A gene. Five hypersensitive sites were located near the transcription initiation site of each promoter in those transgenic-mouse tissues in which the corresponding promoter was active. Only one muscle-specific hypersensitive site was detected, mapping near pM. To functionally delimit the elements required for muscle-specific activity of pM, we performed a deletion analysis of the aldolase A 5' region in transgenic mice. Our results show that a 280-bp fragment containing 235 bp of pM proximal upstream sequences together with the noncoding M exon is sufficient for tissue-specific expression of pM. When a putative MEF-2-binding site residing in this proximal pM region is mutated, pM is still active and no change in its tissue specificity is detected. Furthermore, we observed a modulation of pM activity by elements lying further upstream and downstream from pM. Interestingly, pM was expressed in a tissue-specific way in all transgenic mice in which the 280-bp region was present (32 lines and six founder animals). This observation led us to suggest that the proximal pM region contains elements that are able to override to some extent the effects of the surrounding chromatin.

An opportunistic promoter sharing regulatory sequences with either a muscle-specific or a ubiquitous promoter in the human aldolase A gene.

The human aldolase A gene is transcribed from three different promoters, pN, pM, and pH, all of which are clustered within a small 1.6-kbp DNA domain. pM, which is highly specific to adult skeletal muscle, lies in between pN and pH, which are ubiquitous but particularly active in heart and skeletal muscle. A ubiquitous enhancer, located just upstream of pH start sites, is necessary for the activity of both pH and pN in transient transfection assays. Using transgenic mice, we studied the sequence controlling the muscle-specific promoter pM and the relations between the three promoters and the ubiquitous enhancer. A 4.3-kbp fragment containing the three promoters and the ubiquitous enhancer showed an expression pattern consistent with that known in humans. In addition, while pH was active in both fast and slow skeletal muscles, pM was active only in fast muscle. pM activity was unaltered by the deletion of a 1.8-kbp region containing the ubiquitous enhancer and the pH promoter, whereas pN remained active only in fast skeletal muscle. These findings suggest that in fast skeletal muscle, a tissue-specific enhancer was acting on both pN and pM, whereas in other tissues, the ubiquitous enhancer was necessary for pN activity. Finally, a 2.6-kbp region containing the ubiquitous enhancer and only the pH promoter was sufficient to bring about high-level expression of pH in cardiac and skeletal muscle. Thus, while pH and pM function independently of each other, pN, remarkably, shares regulatory elements with each of them, depending on the tissue. Importantly, expression of the transgenes was independent of the integration site, as originally described for transgenes containing the beta-globin locus control region.

[Influence of marginal and submarginal restoration margins on periodontal tissues]. / Der Einfluss marginaler und submarginaler Restaurationsränder auf das periodontale Gewebe.

Subgingival and irregular restoration margins have an unfavourable influence on the marginal periodont. The margins close to the gingiva of 206 restorations (age of restorations mean = 49.4 months) showed incorrectnesses in most cases with marginal inflammation as a result. That's why the demand of high precision, supragingival positioning of margin restoration and removal of all potential plaque-retentive or mechanic irritated surface is raised.

[The pancreatic toxicity of cyclosporin A. An experimental study in rats]. / Zur Pankreastoxizität von Cyclosporin A. Eine experimentelle Untersuchung an Ratten.

Animal experiments were conducted to elucidate effects of cyclosporine A (CsA) treatment on pancreas. A daily dose of 15 mg/kg B.W. of cyclosporine A was orally administered to recipient animals (rats), over a period of 28 days. Both endocrinic and exocrinic pancreas tissues were morphometrically investigated, using optical light microscopy and electron microscopy. Increase in the exocrinic pancreas portion in response to cyclosporine A treatment was suggested by these morphometric findings. Disorders in the exocrinic pancreas were additionally revealed by electron microscopy. Some of the mitochondria exhibited degenerative alterations, while decline in matrix density as well as separation or rupture of mitochondrial cristae were recorded from others. The assumption might be derived from these results that even therapeutic CsA doses were capable of inflicting disorders on mitochondrial functionally and thus of generating toxic action.

[Construction and managing of maintenance care in periodontology]. / Aufbau und Gestaltung einer Dispensairebetreuung in der Periodontologie.

The maintenance care of periodontal treated patients is the necessary prerequisite for the individual long-life tooth preservation. The recall interval derives from the recidivity, the compliance of the patient, and from the individual disposition. The use of a self-developed data-bank-system relieve evaluation and processing of the extensive data which will be raised site-specific in the recall session. It will be given some directions to the content of the periodontal maintenance care.