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Urinary tract infections (UTIs) caused by anaerobic bacteria have scarcely been reported. Since anaerobic bacteria are commensals of the genitourinary tract, their presence in a urine sample adds ambiguity in making a definitive diagnosis of anaerobic UTI. It is well known that standard urine culture is the gold standard method for the detection, identification, and antimicrobial susceptibility testing of uropathogens. Nonetheless, both the difficulties in establishing them as pathogens and the scarcity of reported anaerobic UTI cases led to the discontinuation of routine urine culture under an anaerobic atmosphere (UCAA). On the other hand, it is important to emphasize that culture-independent methods, such as proteomics and molecular technics, may detect anaerobes directly on a urine sample. Anaerobes are not included in guidelines for the diagnosis and management of UTIs. At the same time, as fastidious uropathogens and antibiotic resistance become more common, accurate pathogen identification becomes even more important for effective UTI treatment. As a result, we conducted a review of the clinical context, pathogen antimicrobial susceptibility, and treatment of patients with anaerobic UTIs. Because UCAA is a contentious topic, we narrowed our search to cases with both negative standard urine culture and positive UCAA.

La rabdomiolisi e definita come la distruzione del muscolo striato. Puo essere traumatica o meno. Da un punto di vista fisiopatologico, si distingue la fase di ischemia muscolare, caratterizzata da acidosi, ipovolemia e insufficienza renale, dalla fase di riperfusione, che aumenta questi fenomeni e contribuisce all’iperkaliemia. La crush syndrome o sindrome da seppellimento rappresenta la causa piu frequente di rabdomiolisi traumatica. Le rabdomiolisi non traumatiche sono legate soprattutto alle abitudini tossiche, alle malattie ereditarie e alle cause iatrogene o infettive. Clinicamente, la sindrome muscolare e caratterizzata da un dolore muscolare associato alla presenza di un disturbo neurologico sensoriale o motorio. La sindrome urinaria, definita da un colore bruno scuro delle urine legato all’esistenza di una mioglobinuria, puo completare un quadro clinico talora piu grezzo. Un tasso di creatina fosfochinasi (CPK) superiore a 1000 UI/l e un segno di rabdomiolisi. L’insufficienza renale si verifica quasi sistematicamente per dei tassi superiori a 75 000 UI/l. La prognosi e legata all’acidosi, all’ipovolemia, all’insufficienza renale e all’iperkaliemia indotta. La gestione terapeutica si propone di correggere e trattare queste anomalie. Si basa, quindi, principalmente sul riempimento vascolare con della soluzione salina isotonica, allo scopo di indurre una diuresi di 3 ml/kg all’ora. Delle terapie adiuvanti, come l’alcalinizzazione o l’osmoterapia, devono essere limitate a delle indicazioni precise (pH

IF 7.191 (2017); International audience; BACKGROUND:Previous studies showed that high and low body mass index (BMI) was associated with worse prognosis in early-stage colorectal cancer (CRC), and low BMI was associated with worse prognosis in metastatic CRC (mCRC). We aimed to assess efficacy outcomes according to BMI.PATIENTS AND METHODS:A pooled analysis of individual data from 2085 patients enrolled in eight FFCD first-line mCRC trials from 1991 to 2013 was performed. Comparisons were made according to the BMI cut-off: Obese (BMI ≥30), overweight patients (BMI ≥ 25), normal BMI patients (BMI: 18.5-24) and thin patients (BMI

Resume Objectif Evaluation de la faisabilite et de l’interet du controle echographique dans la verification du positionnement correct de la sonde nasogastrique (SNG) en prehospitalier. Type d’etude Etude prospective monocentrique observationnelle. Patients et methode Le positionnement correct des SNG de patients intubes en prehospitalier a ete verifie par echographie et confronte systematiquement aux resultats de deux controles prehospitaliers, le test d’insufflation d’air a la seringue combine a l’auscultation epigastrique a la recherche de borborygmes et le test d’aspiration a la seringue de liquide gastrique. Un controle systematique radiologique a ete effectue et compare aux resultats prehospitaliers. Resultats Nous avons inclus 96 patients. L’âge moyen etait de 52 ans (mediane 53,5 ans, ecart-type 23 ans). Dans 83 % des cas ( n = 80), la SNG etait echoreperee instantanement dans la dynamique du geste de pose. Les delais moyens de confirmation echographique du positionnement correcte de la SNG etaient de 7 s (mediane 2 s, ecart-type 16 s) et de 19 s pour les tests a la seringue (mediane 19 s, ecart-type 5 s). Huit controles echographiques ne permettaient pas de visualiser la SNG. Un echoreperage facilite par l’insufflation d’air a la seringue permettait de demasquer la position digestive de la SNG, sans toutefois confirmer sa position gastrique. Le controle radiologique hospitalier confirmait les resultats echographiques prehospitaliers obtenus. Conclusion Le controle echographique du positionnement correct de la sonde nasogastrique dans notre etude est faisable en prehospitalier. Cette technique rapide, non irradiante, plus sensible et specifique que les tests a la seringue couramment utilises en prehospitalier pourrait s’y substituer.

Resume Introduction La douleur est un symptome frequemment rencontre en prehospitalier. Il peut etre mal evalue, sous-estime et parfois mal pris en charge. Patients et methode Il s’agit d’une etude retrospective portant sur l’ensemble des fiches d’interventions medicales ou le mot « douleur » apparait. L’effectif est de 517 patients. L’objectif de notre travail est d’apprecier l’evaluation et la prise en charge de la douleur dans les pathologies medicales et traumatiques. Nous avons denombre deux groupes de patients : traites (T+) et non traites (T−) en fonction de l’evaluation ou non de la douleur. Les medecins ont ete sensibilises au probleme de l’analgesie au travers de reunions de service et de formation medicale continue. Resultats Cent trente-et-un personnes appartiennent au groupe T+ et 386 au groupe T−. Dans le groupe T+, l’etiologie traumatique represente 71 % des cas et l’evaluation douloureuse est effectuee dans 55 % des cas. Dans le groupe T−, l’etiologie medicale represente 91 % des cas et l’evaluation douloureuse est effectuee dans 13 % des cas. L’etiologie traumatique et l’âge d’autant plus jeune sont des facteurs favorisant la prise en charge de la douleur par le medecin. Conclusion Notre travail souligne l’insuffisance d’evaluation et de traitement de la douleur dans notre service. La sensibilisation de l’ensemble de l’equipe soignante est indispensable. La formation medicale continue, la diffusion d’un protocole d’analgesie, l’utilisation d’echelles d’evaluation adaptees et l’investissement de l’infirmier dans l’evaluation douloureuse prennent toute leur importance afin d’ameliorer la prise en charge des patients.

Renin-angiotensin system antagonists, either angiotensin-converting enzyme (ACE) inhibitors or angiotensin II receptor (AT(1)) antagonists, may interfere with regulation of arterial pressure during anaesthesia. This study aimed to compare the haemodynamic profile of anaesthetized pigs, which were subjected to haemorrhage in the presence of the ACE inhibitor enalaprilat or the AT(1) antagonist valsartan.Thirty-six pigs were assigned randomly to placebo, enalaprilat or valsartan groups. After a 30-min period of stabilization following anaesthesia and injection of the study drug, the animals were bled in two equal steps of 20% of their estimated blood volume (20% BV and 40% BV).After bleeding of 20% BV, the mean arterial pressure (MAP) decreased significantly but similarly in each group (20-25%) but the placebo and the enalaprilat groups had a significant decrease in cardiac index (CI, 22% and 16%, respectively) without significant change in systemic vascular resistance (SVR). Conversely, in the valsartan group, SVR decreased significantly (23%, P0.02 vs other groups) without significant change in CI (-4%). After bleeding of 40% BV, the CI decreased significantly compared with 20% BV in the three groups (19% in the placebo and enalaprilat groups, 14% in the valsartan group) but the MAP decreased significantly in the enalaprilat group only (23%). The SVR increased significantly in the placebo group (P0.01 vs each of the other groups), but there were no differences in the change in SVR between the other groups.Blockade of the renin-angiotensin system by either enalaprilat or valsartan leads to a similar decrease in arterial pressure during anaesthesia and haemorrhage but the haemodynamic profiles are quite different.

PURPOSE: One of the major challenges for health care professionals in heart failure (HF) management is to maintain and/or improve HF patient health-related quality of life. The Minnesota Living With Heart Failure Questionnaire (MLHFQ) is one of the most comprehensive and widely used tools for measuring health-related quality of life among patients with HF. The aim of the study was to assess the psychometric properties of the Greek version of the tool. METHODS: The MLHFQ was administered among 128 Greek-Cypriot HF patients to assess the internal consistency, content validity, and contrast validity of its Greek version. Exploratory factor analysis was undertaken to establish its construct validity. RESULTS: The factor analysis in this study provided support for a 3-factor solution explaining 64.15% of the variance (physical, emotional, and social subscales). The internal consistency for the Greek version of the MLHFQ total scale (0.95) and subscales (0.80-0.94) were found to be high. The contrast validity of the Greek version of the MLHFQ was explored through cumulative MLHFQ scores and comparisons that were able to distinguish among all different levels of HF severity, as defined by the New York Heart Association functional class grouping. CONCLUSION: This study provides support for the reliability and validity of the Greek version of the MLHFQ. Copyright © 2013 Wolters Kluwer Health | Lippincott Williams &Wilkins.

Autoradiographic experiments using iodinated vasopressin analog revealed the presence of specific vasopressin-binding sites in the human adrenal cortex (zona glomerulosa and zona fasciculata). These receptors exhibited a good affinity for arginine vasopressin (3.3 nM), with classical V1a pharmacology and densities of 65 and 135 fmol/mg protein-enriched membranes from zona glomerulosa and fasciculata, respectively. Vasopressin receptors present in both glomerulosa and fasciculata cell-enriched primary cultures were coupled to phospholipase C (ED50, 0.9 and 1.8 nM; maximal stimulation, 4.3- and 5.8-fold, respectively). Vasopressin also stimulated an increase in intracellular calcium through at least two distinct mechanisms: the mobilization of intracellular pools via vasopressin-stimulated inositol phosphate accumulation and the activation of calcium influx. In glomerulosa cell-enriched primary cultures, vasopressin increased aldosterone secretion (ED50, 0.4 nM; maximal stimulation, 2.5-fold) and was found to be as potent as angiotensin-II in stimulating aldosterone secretion, phosphoinositide turnover, and calcium mobilization. In fasciculata cells, vasopressin and angiotensin-II were also able to stimulate cortisol secretion and inositol phosphate accumulation. Moreover, perifusion experiments demonstrated that vasopressin was released from the adrenal medulla. Together, these results indicate that vasopressin can be considered a potent paracrine modulator of adrenal steroid secretion in man.

Decision-makers increasingly need to bring together multiple models across a broad range of disciplines to guide investment and policy decisions around highly complex issues such as population health and safety. We discuss the use of the Smarter Planet Platform for Analysis Simulation of Health (Splash) for cross-disciplinary modeling, simulation, sensitivity analysis, and optimization in the setting of complex systems of systems. Splash is a prototype system that allows combination of existing heterogeneous simulation models and datasets to create composite simulation models of complex systems. Splash, built on a combination of data-integration, workflow management, and simulation technologies, facilitates loose coupling of models via data exchange. We describe the various components of Splash, with an emphasis on the experiment-management component. This latter component uses user-supplied metadata about models and datasets to provide, via an interactive GUI, a unified view over all of the parameters in all of the component models that make up a composite model, a mechanism for selecting the factors to vary, and a means for allowing users to easily specify experimental designs for the selected factors. The experiment manager also provides a mechanism for systematically varying the inputs to the composite models. We show how the experiment manager can be used to implement some simple stochastic-optimization functionality by implementing the Rinott procedure for selecting the best system. We also implement a sensitivity-analysis method based on a fractional-factorial experimental design. We demonstrate this technology via a composite model comprising a financial-rate model and a healthcare payer model.

To assess the feasibility and actual performance of ultrasound control in verification of the correct positioning of a nasogastric tube in pre-hospital settings.Prospective, observational, single-centre study.Correct positioning of nasogastric tubes in patients intubated in a pre-hospital setting was verified by ultrasound and routinely compared with the results of two pre-hospital tests, namely a test involving insufflation of air through a syringe coupled with epigastric auscultation and a test involving aspiration of gastric fluid with a syringe. Routine x-ray control was carried out and compared with the pre-hospital results.Ninety-six patients were included. Mean age was 52 years (median: 53.5 years, SD: 23 years). In 83% of the patients (n=80), the nasogastric tube was located by ultrasound immediately during the insertion procedure. The mean times to ultrasound confirmation of correct positioning of the nasogastric tube were 7s (median: 2s; SD: 16s) and 19s for the syringe tests (median 19s, SD: 5s). Eight ultrasound control tests were negative. Location coupled with insufflation of air through a syringe allowed detection of the nasogastric tube in the stomach but without providing confirmation of the actual gastric position. The pre-hospital ultrasound results were confirmed by subsequent radiological controls at the hospital.The ultrasound test performed in our study to verify correct positioning of a nasogastric tube is feasible in a pre-hospital setting. This technique is rapid and non-irradiating and is more sensitive and specific than the syringe tests commonly used in pre-hospital settings, and it may be performed in place of the latter tests.

We show that prospect theory offers a rich theory of casino gambling, one that captures several features of actual gambling behavior. First, we demonstrate that, for a wide range of preference parameter values, a prospect theory agent would be willing to gamble in a casino even if the casino only offers bets with no skewness and with zero or negative expected value. Second, we show that the probability weighting embedded in prospect theory leads to a plausible time inconsistency: at the moment he enters a casino, the agent plans to follow one particular gambling strategy; but after he starts playing, he wants to switch to a different strategy. The model therefore predicts heterogeneity in gambling behavior: how a gambler behaves depends on whether he is aware of the time inconsistency; and, if he is aware of it, on whether he can commit in advance to his initial plan of action.

A number of authors have suggested that investors derive utility from realizing gains and losses on assets that they own. We present a model of this "realization utility," analyze its predictions, and show that it can shed light on a number of puzzling facts. These include the disposition effect, the poor trading performance of individual investors, the higher volume of trade in rising markets, the effect of historical highs on the propensity to sell, the individual investor preference for volatile stocks, the low average return of volatile stocks, and the heavy trading associated with highly valued assets.

Enterobacter spp. are becoming increasingly frequent nosocomial pathogens with multiple resistance mechanism to beta-lactam antibiotics. We carried out the phenotypic and genotypic characterization of beta-lactamases in 27 Enterobacter spp. (25 Enterobacter cloacae y 2 Enterobacter aerogenes), as well as the ability of different extended spectrum-lactamase (ESBL) screening methods. Resistance to third generation cephalosporins was observed in 15/27 (63%) isolates. Twelve resistant isolates produced high level chromosomal encoded AmpC beta-lactamase; 6 of them were also producers of PER-2. Resistance to third generation cephalosporins in the remaining 3 isolates was due to the presence of ESBLs, PER-2 in 2 cases, and CTX-M-2 in the other. Only CTX-M-2 production was detected with all tested cephalosporins using difusion synergy tests, while cefepime improved ESBLs detection in 7/8 PER-2 producers, 4/8 in the inhibitor approximation test and 7/8 with double disk test using cefepime containing disk with and without clavulanic acid. Dilution method, including cephalosporins with and without the inhibitor detected 1/9 ESBLs producers.

Eikenella corrodens is a gram-negative bacillus that colonizes as normal flora of the mouth, the upper respiratory tract and the gastrointestinal tract. The aim of this study was to determine the susceptibility patterns against fourteen antibiotics of 25 E. corrodens strains isolated at our hospital. MICs were determined by the agar dilution technique using Müeller-Hinton agar with sheep blood (5% v/v) to penicillin, ampicillin, ampicillin-sulbactam, cephalotin, cefoxitin, ceftiaxone, colistin, gentamicin, amikacin, erythromycin, rifampin, ciprofloxacin and clindamycin. The most active antibiotics were ciprofloxacin and ceftriaxone (MIC90 = 0.008 and 0.125 microgram/ml, respectively), whereas eritromycin, gentamicin and amikacin showed less activity. Only one strain was beta lactamase positive, and it was inhibited by sulbactam. Erithromycin, gentamicin and amikacin had poor activity (MIC90 = 16.8 and 64 micrograms/ml, respectively), whereas all the strains were uniformly resistant to clindamycin (MICor = 32 micrograms/ml). We suggest about the need of periodical surveys of E. corrodens susceptibility patterns, since strains have been found with decreased susceptibility against antibiotics which are currently being used for the treatment of infectious diseases.

Reliable data regarding the efficacy of different schemes of triple therapy for the eradication of Helicobacter pylori in our country, are not available. Patients with Helicobacter pylori infection and non-ulcer dyspepsia or active peptic ulcer disease were randomized in three different groups for therapy with, omeprazole 20 mg, clarithromycin 500 mg and amoxicillin 1000 mg, twice daily for one week (OCA 1, 40 patients) and the same treatment but for two weeks in a second group (OCA 2, 40 patients). The third group received omeprazole 20 mg, clarithromycin 500 mg and metronidazole 500 mg twice daily during one week (OCM, 40 patients). The primary efficacy end point was the eradication of Helicobacter pylori as confirmed by negative urea breath test, 4 weeks after the completion of treatment. Of 120 patients enrolled in the study, 113 met the entry criteria. Of them, 103 completed the treatment. When analyzed by intention to treat, after 4 weeks of finishing the treatment, Helicobacter pylori was eradicated in 92.3% of patients in OCA 1, 89.7% in OCA 2, and 82.8% in OCM. There was no significant difference between the three groups, regarding the eradication efficacy. Side effects were observed more frequently in OCA 2 and OCM groups. Primary resistance to amoxicillin and clarithromycin was not demonstrated, while 20% of cultured strains were resistant to metronidazole. In patients with peptic ulcer disease or non-ulcer dysplasia, triple therapy with omeprazole and two antibiotics is highly effective in the eradication of Helicobacter pylori. One week of OCA therapy is as effective as two weeks of OCA or one week of OCM, with less side effects.

Understanding of the molecular determinants responsible for antagonist binding to the oxytocin receptor should provide important insights that facilitate rational design of potential therapeutic agents for the treatment of preterm labor. To study ligand/receptor interactions, we used a novel photosensitive radioiodinated antagonist of the human oxytocin receptor, d(CH(2))(5) [Tyr(Me)(2),Thr(4),Orn(8),Phe(3(125)I,4N(3))-NH(2)9]vasotocin. This ligand had an equivalent high affinity for human oxytocin and V(1a) vasopressin receptors expressed in Chinese hamster ovary cells. Taking advantage of this dual specificity, we conducted photoaffinity labeling experiments on both receptors. Photolabeled oxytocin and V(1a) receptors appeared as a unique protein band at 70-75 kDa and two labeled protein bands at 85-90 and 46 kDa, respectively. To identify contact sites between the antagonist and the receptors, the labeled 70-75- and the 46-kDa proteins were cleaved with CNBr and digested with Lys-C and Arg-C endoproteinases. The fragmentation patterns allowed the identification of a covalently labeled region in the oxytocin receptor transmembrane domain III consisting of the residues Leu(114)-Val(115)-Lys(116). Analysis of contact sites in the V(1a) receptor led to the identification of the homologous region consisting of the residues Val(126)-Val(127)-Lys(128). Binding domains were confirmed by mutation of several CNBr cleavage sites in the oxytocin receptor and of one Lys-C cleavage site in the V(1a) receptor. The results are in agreement with previous experimental data and three-dimensional models of agonist and antagonist binding to members of the oxytocin/vasopressin receptor family.

Despite their opposite effects on signal transduction, the nonapeptide hormone arginine-vasopressin (AVP) and its V1a receptor-selective cyclic peptide antagonist d(CH2)5[Tyr(Me)2]AVP display homologous primary structures, differing only at residues 1 and 2. These structural similarities led us to hypothesize that both ligands could interact with the same binding pocket in the V1a receptor. To determine receptor residues responsible for discriminating binding of agonist and antagonist ligands, we performed site-directed mutagenesis of conserved aromatic and hydrophilic residues as well as nonconserved residues, all located in the transmembrane binding pocket of the V1a receptor. Mutation of aromatic residues of transmembrane region VI (W304, F307, F308) reduced affinity for the d(CH2)5[Tyr(Me)2]AVP and markedly decreased affinity for the unrelated strongly hydrophobic V1a-selective nonpeptide antagonist SR 49059. Replacement of these aromatic residues had no effect on AVP binding, but increased AVP-induced coupling efficacy of the receptor for its G protein. Mutating hydrophilic residues Q108, K128 and Q185 in transmembrane regions II, III and IV, respectively, led to a decrease in affinity for both agonists and antagonists. Finally, the nonconserved residues T333 and A334 in transmembrane region VII, controlled the V1a/V2 binding selectivity for both nonpeptide and cyclic peptide antagonists. Thus, because conserved aromatic residues of the V1a receptor binding pocket seem essential for antagonists and do not contribute at all to the binding of agonists, we propose that these residues differentiate agonist vs. antagonist ligand binding.

A novel photoactivatable linear peptide antagonist selective for the V(1a) vasopressin receptor, [(125)I][Lys(3N(3) Phpa)(8)]HO-LVA, was synthesized, characterized, and used to photolabel the human receptor expressed in Chinese hamster ovary cells. Two specific glycosylated protein species at 85-90 and 46 kDa were covalently labeled, a result identical to that obtained with a previous photosensitive ligand, [(125)I]3N(3)Phpa-LVA (Phalipou, S., Cotte, N. , Carnazzi, E., Seyer, R., Mahe, E., Jard, S., Barberis, C., and Mouillac, B. (1997) J. Biol. Chem. 272, 26536-26544). To identify contact sites between the new photoreactive analogue and the V(1a) receptor, the labeled receptors were digested with Lys-C or Asp-N endoproteinases and chemically cleaved with CNBr. Fragmentation with CNBr, Lyc-C, and Asp-N used alone or in combination, led to the identification of a restricted receptor region spanning the first extracellular loop. The results established that sequence Asp(112)-Pro(120) could be considered as the smallest covalently labeled fragment with [(125)I][Lys(3N(3)Phpa)(8)]HO-LVA. Based on the present experimental result and on previous photoaffinity labeling data obtained with [(125)I]3N(3)Phpa-LVA (covalent attachment to transmembrane domain VII), three-dimensional models of the antagonist-bound receptors were constructed and then verified by site-directed mutagenesis studies. Strikingly, these two linear peptide antagonists, when bound to the V(1a) receptor, could adopt a pseudocyclic conformation similar to that of the cyclic agonists. Despite divergent functional properties, these peptide antagonists could interact with a transmembrane-binding site significantly overlapping that of the natural hormone vasopressin.

Tuberculous cervicitis (TC) is a rare disease the diagnosis of which is based on the microscopic and/or cultural recognition of mycobacteria. In recent years, the polymerase chain reaction (PCR), especially with double-round amplification ("nested" PCR [nPCR]), has been increasingly used for rapid detection of mycobacteria in clinical samples.The present case is the first example of tuberculosis diagnosed with the aid of nPCR amplification of mycobacterial DNA fragments on smeared and Papanicolaou-stained cytologic material. First detected on vaginal smears, the amplicon IS6110 was subsequently identified also on paraffin-embedded tissue sections.The technique described here could also be applied to aspiration cytology smears to give rapid and accurate information on mycobacterial infections.

High affinity binding sites for vasopressin (VP) are widely distributed within the rat brain and spinal cord. Since their presence is associated with neuronal sensitivity to VP application, their anatomical distribution maps structures which could be activated by endogenous VP. Interestingly, marked species-related differences of the VP receptor distribution have been revealed. Some evidence has also been provided that mechanisms of receptor regulation may vary among species. In the rat, the expression of VP binding sites in some motor nuclei shows remarkable plasticity, in particular up-regulation after axotomy. These data suggest that VP may, in addition to affecting motoneuronal excitability, act as a trophic factor onto motoneurones.

An unusual campylobacter strain, a urease positive thermophilic variant of Campylobacter lari, was isolated from the urine of a patient with urinary tract infection who was hospitalized because of cirrhosis and haemorrhage. The strain was isolated from urine specimens on three separate occasions. A significant serological response to the organism was also detected. This is the first documented case of extra-intestinal infection due to this group of organisms.

To document that a polyclonal antiserum to calretinin, a 29-kd calcium-binding protein, consistently decorates normal and tumor mesothelial cells in cytologic preparations.Thirty-three archival cytologic specimens from eight patients with histologically confirmed malignant mesothelioma and 13 from patients with metastatic serous effusions were destained and then immunostained with anticalretinin antiserum. For investigation of cell suspensions, four pleural fluids were incubated with anticalretinin antiserum. After cytocentrifugation the specimens were stained in accordance with the alkaline phosphatase anti-alkaline phosphatase (APAAP) method. For electron microscopic examination the cell suspensions were then incubated with gold-labeled antirabbit antibody.The diagnostic sensitivity of this new immunocytochemical approach reached 100% for the eight malignant mesotheliomas investigated. Only 3 of the 13 adenocarcinomas metastatic to the serous membranes included in this study were weakly reactive, accounting for 81% specificity. Binding of anticalretinin antiserum to living mesothelial cells was consistently documented in all four cases investigated.Calretinin is a very useful marker for positive identification of normal and tumor mesothelial cells in serous effusions.

To identify receptor functional domains underlying binding of the neurohypophysial hormones vasopressin (AVP) and oxytocin (OT), we have constructed a three-dimensional (3D) model of the V1a vasopressin receptor subtype and docked the endogenous ligand AVP. To verify and to refine the 3D model, residues likely to be involved in agonist binding were selected for site-directed mutagenesis. Our experimental results suggest that AVP, which is characterized by a cyclic structure, could be completely buried into a 15-20-A deep cleft defined by the transmembrane helices of the receptor and interact with amino acids located within this region. Moreover, the AVP-binding site is situated in a position equivalent to that described for the cationic neurotransmitters. Since all mutated residues are highly conserved in AVP and OT receptors, we propose that the same agonist-binding site is shared by all members of this receptor family. In contrast, the affinity for the antagonists tested, including those with a structure closely related to AVP, is not affected by mutations. This indicates a different binding mode for agonists and antagonists in the vasopressin receptor.

An established clinical development plan was expanded to form a framework for a competency assessment model. Each nursing competency has a technical, interpersonal and critical thinking component. Behaviors describe all areas from novice to expert. The framework identifies and supports a professional model of nursing practice.

Despite intensive efforts over three decades in many laboratories, attempts to design peptide antagonists of oxytocin (OT) which are more selective for OT uterine receptors than for vasopressin (AVP), vasopressor V1a receptors, have met with only limited success. We will review the current status of the field and report on studies in our laboratories which have led to the design of highly potent non-selective and selective OT antagonists. Virtually all are more potent (2-6 fold) and a number are more selective (10-12 fold) than Atosiban, currently in clinical trial as a tocolytic agent. Many of these new published and unpublished OT antagonists are thus promising candidates for development as potential tocolytic agents for the prevention of pre-term labor. We also report on promising new radioiodinatable ligands for OT receptors. All the new OT antagonists are valuable new tools for studies on the physiological roles of OT and as probes for OT and AVP receptors.

The present study aims at delineating residues in the vasopressin/oxytocin receptor family responsible for the high affinity binding of the hormone. Therefore, we have constructed a computer-generated 3 dimensional model of the rat V1a vasopressin receptor subtype which allowed us to propose residues likely to be involved in agonist binding. Among these residues, several are highly conserved in the receptor family. They were selected for site-directed mutagenesis on the basis of putative direct interaction with bound ligands. The present model and experimental results led us to conclude that the hormone is docked in a pocket completely buried in the transmembrane core of the receptor. Large polar residues, such as glutamine and lysine, located in transmembrane regions 2,3,4 and 6 are involved in the binding of the neurohypophysial hormone. Since all the mutated residues are highly conserved in AVP and OT receptors, we propose that the agonist binding site is similar in all members of the receptor family; only minor changes were found in antagonist potencies, suggesting that agonist and antagonist binding sites do not completely overlap.

Using autoradiography on film, specific binding sites for arginine-vasopressin (AVP) and for oxytocin (OT) were localized in various areas of the brain of adult male guinea pigs. Vasopressin binding sites were detected with [3H]AVP or with [125I]VPA, a recently synthetized linear vasopressin antagonist radiolabeled with 125I. [125I]VPA and [3H]AVP yielded similar results, thus suggesting that AVP binding sites present in the guinea pig brain are V1 type receptors. Supporting evidence on this was obtained in competing studies using structural analogues allowing to discriminate V1 receptors from V2 and from OT receptors. Oxytocin binding sites were labeled with [3H]OT or with the iodinated OT antagonist [125I]OTA; both ligands yielded similar results. The localization in the guinea pig brain of AVP binding sites differed from that of OT binding sites. AVP binding sites were mainly detected in the olfactory bulb and throughout the cerebral cortex. Oxytocin binding sites were most noticeable in the hypothalamic ventromedial nucleus, in the amygdaloid complex and in restricted areas of the cerebral cortex. A comparison of the present data with those previously described in the rat, the mouse, the human and the hamster brain suggests that similar binding sites are present in these species, but that their anatomical distribution differs markedly. These data are discussed in relation to immunocytochemical and electrophysiological data which suggest that binding sites detected by autoradiography may represent, at least in part, functional neuronal receptors.

A cDNA of 2149 base pairs with an incomplete open reading frame (ORF) encoding amino acids 1-516 of a 531-amino acid protein highly homologous to muscarinic receptors was cloned from a rat brain cDNA library. The complete ORF was then deduced from a DNA fragment cloned from a rat genomic library. This ORF was subcloned into the eukaryotic expression vector p91023(B) under control of the adenovirus major late promoter and co-transfected with the thymidine kinase selection marker into muscarinic receptor-negative, thymidine kinase-negative murine L cells. Stable transformants were selected and tested for acquisition of muscarinic receptors by following appearance of specific binding sites for the muscarinic ligand [3H] N-methylscopolamine. Two cell lines, LM5.36 and LM5.40, were cloned and shown to express typical muscarinic receptor sites, thus confirming that the newly cloned ORF encodes a muscarinic receptor, the rat M5 muscarinic acetylcholine receptor. Tests for activities showed it to stimulate phosphoinositide hydrolysis in intact cells, without affecting positively or negatively adenylyl cyclase activity. The M5 receptor contains two putative glycosylation sites at its amino terminus and, based on hydropathicity analysis, is predicted to span the plasma membrane seven times. Like 17 other receptors of this class, the M5 receptor has 19 conserved amino acids, among which are 4 prolines located in the 4th, 5th, 6th, and 7th predicted transmembrane regions, conferring possible bends to these helices, and 2 cysteines, one in the 1st and the other in the 2nd extracellular loop, possibly providing for a disulfide bond. Similarity in amino acid composition and in patterns of antagonist binding and biologic effects suggest the M5 receptor to be M1-like.

The distribution of cytochrome c oxidase, monoamine oxidase and lactate dehydrogenase, together with protein, after isopycnic centrifugation of a crude mitochondrial fraction of chick telencephalon homogenate in a linear sucrose density gradient, was followed during late embryogenesis and postnatal maturation. Two main populations of subcellular organelles differentiate; they were characterized biochemically and analyzed by electron microscopy. One population, with a progressively defined mean bouyant density of 1.170 g/ml, exhibited a high relative activity of monoamine oxidase, with a low and relatively constant cytochrome c oxidase/monoamine oxidase activity ratio. This population was composed of free mitochondria and mitochondria enclosed in nerve endings, and possibly of mitochondria of perikaryal and glial origin. A second population, with a progressively well defined mean buoyant density of 1.182–1.186 g/ml, exhibited a high relative activity of cytochrome c oxidase, with a high and increasing cytochrome c oxidase/monoamine oxidase activity ratio. The biochemical and functional significance of these results were discussed.

— –(l) The contents of potassium and of ATP, ADP and AMP of homogenates of guinea pig neocortex in 0.32 M-SUCROSE, and of synaptosomal preparations derived therefrom, were determined and effects of incubation, superfusion and stimulation of the preparation were examined. The synaptosome preparations in M-SUCTOSe carried a smaller content of 5′-nucleotide per unit protein than did the homogenate from which they were derived. However, the proportion of synaptosomal nucleotides present as ATP was markedly greater than in the homogenate as a whole, the adenylate energy-charge being 83% greater in the synaptosomes than in the homogenate. Dilution of the synaptosomal preparation from the 1 M-sucrose to isotonic sucrose, decreased the contcnt of ATP per unit protein, but did not change the sum ATP + ADP + AMP. (2) Examined as deposited beds during 5 to 20 min incubation in oxygenated glucose-bicarbonate salines, synaptosomal K content and adenylate energy charge increased. These changes were sustained during a subsequent 40 min of incubation and also during superfusion. During such continued superfusion, electrical stimulation caused diminution of the ATP and the adenylate energy charge of the beds, as also did superfusion with fluids of increased K-content. (3) Lactate formation by the superfused beds was relatively stable during 30 min incubation. By electrical stimulation, the rate of lactate formation was increased by up to 30%. Increase in the potassium content of superfusion fluids could however increase lactate production 2.4-fold. The basis for these actions is discussed.

A cDNA of 2149 base pairs with an incomplete open reading frame (ORF) encoding amino acids 1-516 of a 531-amino acid protein highly homologous to muscarinic receptors was cloned from a rat brain cDNA library. The complete ORF was then deduced from a DNA fragment cloned from a rat genomic library. This ORF was subcloned into the eukaryotic expression vector p91023(B) under control of the adenovirus major late promoter and co-transfected with the thymidine kinase selection marker into muscarinic receptor-negative, thymidine kinase-negative murine L cells. Stable transformants were selected and tested for acquisition of muscarinic receptors by following appearance of specific binding sites for the muscarinic ligand [3H] N-methylscopolamine. Two cell lines, LM5.36 and LM5.40, were cloned and shown to express typical muscarinic receptor sites, thus confirming that the newly cloned ORF encodes a muscarinic receptor, the rat M5 muscarinic acetylcholine receptor. Tests for activities showed it to stimulate phosphoinositide hydrolysis in intact cells, without affecting positively or negatively adenylyl cyclase activity. The M5 receptor contains two putative glycosylation sites at its amino terminus and, based on hydropathicity analysis, is predicted to span the plasma membrane seven times. Like 17 other receptors of this class, the M5 receptor has 19 conserved amino acids, among which are 4 prolines located in the 4th, 5th, 6th, and 7th predicted transmembrane regions, conferring possible bends to these helices, and 2 cysteines, one in the 1st and the other in the 2nd extracellular loop, possibly providing for a disulfide bond. Similarity in amino acid composition and in patterns of antagonist binding and biologic effects suggest the M5 receptor to be M1-like.

A new, highly selective radio-iodinated oxytocin receptor antagonist [( 1-(beta-mercapto-beta, beta-cyclopentamethylenepropionic acid, 2-O-methyltyrosine, 4-threonine, 8-ornithine, 9-tyrosylamide]-vasotocin) was used to identify and quantitate specific binding sites for the neurohypophyseal hormone oxytocin with in vitro incubation of rat brain sections and autoradiography. Exclusively oxytocin binding sites were detected in view of the high affinity of the [125I]-labelled oxytocin antagonist for oxytocin binding sites and the negligible affinity for the vasopressin liver (V1) and kidney (V2) receptor types. The putative oxytocin receptors were abundantly present in several brain regions, where previously discrimination between oxytocin and vasopressin binding was difficult, i.e. the olfactory nucleus, the islands of Calleja, the ventromedial nucleus of the hypothalamus, the central amygdaloid nucleus and the ventral subiculum of the hippocampus. In addition oxytocin receptors were demonstrated in other areas, such as the taenia tecta, dorsolateral caudate putamen, ventral pallidum, accumbens, lateral septum, bed nucleus of the stria terminalis, thalamic paraventricular nucleus, lateral, basolateral and medial amygdala, the dorsal subiculum, perirhinal cortex and the amygdaloid-hippocampal area. The high affinity and the low detection threshold of this [125I]-labelled oxytocin antagonist permitted identification of oxytocin receptors in new regions such as the ventral part of the lateral septum, medial septum, dorsal motor nucleus of the vagus nerve and the olive nuclei in the brain stem.

Kinetics for transport of adenosine into guinea-pig neocortex synaptosomes were studied by incubating them with [14C]adenosine for up to 30 s. The apparent Km value of the high-affinity transport system for adenosine was 21.1 microM and the Vmax value was 257.3 pmol/min/mg protein. The transport system was inhibited by both compounds structurally related (compounds 554 and 555) and unrelated (dipyridamole) to adenosine. Because electrically stimulated synaptosomes release up to 1.5% of the adenosine derivative content per min, the physiological significance of adenosine uptake is discussed as a possible mechanism to compensate for the loss of adenine nucleotides from synaptosomal preparations.