Survival and inactivation of hepatitis E virus on inanimate surfaces.

BACKGROUND: Hepatitis E virus (HEV) is the most common cause of acute viral hepatitis, and mainly transmitted via faecal-oral contamination or consumption of contaminated food products. However, limited data on the surface stability and HEV sensitivity to chemical disinfectants are available. AIM: To establish an HEV-based carrier assay to evaluate its surface stability and the virucidal activity of nine surface disinfectants. METHODS: A recently developed robust HEV-3 cell culture system for an HEV-based carrier assay. FINDINGS: Alcohol-based disinfectants were insufficient to eliminate HEV infectivity, whereas disinfectants based on aldehyde, peracetic acid, oxygen, and/or quaternary ammonium inactivated HEV. CONCLUSION: These findings have strong implications for the recommendation of evidence-based hygiene guidelines to reduce HEV transmission.

Single-catheter approach to pulmonary vein reisolation in selected patients : Data from a prospective registry.

BACKGROUND: In addition to different types of single-tip ablation catheters for pulmonary vein (PV) reisolation, a newly developed circular mapping and ablation catheter (nMARQ®) has been available since 2013 and is currently used only in initial PV isolation procedures. In this prospective registry we present feasibility and efficacy data for PV reisolation procedures with a single-catheter approach (nMARQ®) compared with a standard approach using a single-tip ablation catheter and a circular mapping catheter. METHODS: We included 35 carefully selected patients in this prospective registry and assigned them in a 2:1 ratio to undergo either PV reisolation with a single-tip ablation catheter together with a steerable circular mapping catheter (group 1) or with the nMARQ®catheter only (group 2). The recurrence rate was calculated for atrial tachyarrhythmias with a duration of > 30 s during a mean follow-up of 12.7 months. RESULTS: Reisolation of all PVs was achieved in all patients of both groups. In group 2, all gaps could be correctly identified with the nMARQ® catheter. PV isolation was clearly visible on the nMARQ® catheter in all targeted veins. With the nMARQ® catheter the ablation time decreased significantly (6.3 ± 3.0 vs. 18.6 ± 13.9 min, p < 0.05). The recurrence rate of atrial fibrillation did not differ significantly between the two groups (37.5 vs. 45.5 %, p = 0.66). CONCLUSION: In selected patients, a complete PV reisolation procedure is feasible with a singular circular mapping and ablation catheter. The 12-months success rate is comparable to a classic approach with a combination of a single-tip ablation catheter and a circular mapping catheter.

Nursing students' use of a psychopharmacology game for client empowerment.

Theoretical support is evaluated for nursing students' use of an innovative educational intervention, the Psychopharmacology R.A.C.E. (Wissman and Tankel, Kansas City, MO) game, as an empowerment tool with mental health clients. Based on mutual trust and respect, the interpersonal process occurs between an individual or group (e.g., client, student) with a desire for competency growth (e.g., knowledge, proactive health behaviors, negotiation), and development of a sense of control and self-determination and another individual or group (e.g., nurse) who serves as an enabler, supporter, and resource mobilizer. A review of empowerment literature identifies: (1) support of education as an empowerment tool; (2) identification of specific teaching methods and behaviors that promote empowerment; (3) empowerment as an outcome of involvement; and (4) support of the empowerment value of group participation. Preliminary clinical observations reveal strong support for nursing students' use of the Psychopharmacology R.A.C.E. game as a method to empower mental health clients.

Engineering of substrate mimetics as novel-type substrates for glutamic acid-specific endopeptidases: design, synthesis, and application.

This account reports on the development and function of novel substrate mimetics as artificial substrates for Glu-specific endopeptidases. Firstly, in an empirical way, various aliphatic and aromatic analogs of the already established carboxymethyl thioester-substrate mimetics were designed from simple structure-function relationship studies. The specificity of the newly developed substrates for Staphylococcus aureus V8 protease-catalyzed reactions have been examined by steady-state hydrolysis kinetic studies. Additionally, these studies were expanded to the use of the equally Glu-specific endopeptidase from Bacillus licheniformis (BL-GSE) which can easily be purified from alcalase in high yields. Finally, the novel substrate mimetics were used as acyl donor components in BL-GSE- and V8 protease-catalyzed model acyl transfer reactions. The results clarify the newly developed substrate mimetics as efficient acyl donors as well as BL-GSE as an attractive alternative to V8 protease for enzymatic peptide synthesis.

Substrate mimetics and freezing strategy: a useful combination that broadens the scope of proteases for synthesis.

[reaction: see text] We present an irreversible and efficient protease-based method for peptide synthesis which occurs independently of the primary specificity of proteases and also without proteolytic side reactions. The key feature of this approach is the combination of the substrate mimetics strategy with frozen state enzymology. Model reactions catalyzed by several proteases qualify this approach as a powerful concept in the direction of a more universal application of proteases as biocatalysts for peptide ligation.

Psychopharmacology RACE: a fun road to competence.

1 In this era of rapidly changing mental health services and new psychotropic drugs, health professionals are challenged to develop and maintain psychopharmacology competencies. 2 The Psychopharmacology RACE provides an interactive and learner-oriented method of developing and assessing theoretical and clinical competencies for psychopharmacology. 3 The Psychopharmacology RACE is also well suited for modification and application toward the professional development of nurses and the education of clients.

Reverse catalysis of elastase from porcine pancreas in frozen aqueous systems.

The reverse action of a trypsin-free elastase isolated from porcine pancreas was studied in frozen aqueous systems. Under frozen state conditions, porcine pancreatic elastase was able to catalyse peptide bond formation more effectively than in solution at room temperature. The acceptance of free amino acids as nucleophilic amino components indicates a changed specificity of the endoprotease in frozen reaction mixtures. In elastase-catalysed formation of Ser-, Ile- and Val-X-bonds in frozen aqueous reaction mixtures, peptide yields obtained depended on the P1 amino acid and the acyl donor chain length.

Annexins in Paramecium cells. Involvement in site-specific positioning of secretory organelles.

Annexins were isolated from Paramecium cell homogenates by standard ethylene glycol tetraacetic acid (EGTA) extraction and 100 000-g centrifugation. Two different antibodies (Abs) against synthetic peptides were used, Call-15 and B15, which in mammalian cells recognize a sequence of annexin II or a common sequence occurring in several annexins (except for annexin II), respectively. With anti-Call-15 Abs, western blots from EGTA extracts showed strongly reactive bands of 44.5 and 46 kDa and of higher values. Some of these bands bound to the 100 000-g pellet fraction when Ca(2+) was added. Immuno- and affinity labelling revealed selective, Ca(2+)-dependent labelling of the cell cortex, with enrichment around trichocyst docking sites (facing subplasmalemmal Ca(2+) stores). Cortical fluorescence labelling decreased in wild-type (7S) cells when trichocyst ghosts were detached after synchronous exocytosis. Similarly, cortical labelling was reduced when intact trichocysts were detached from the cell surface of non-discharge mutant cells (nd9-28 degrees C, showing identical bands on blots), which then contained numerous heavily labelled phagolysosomes. This strongly suggests annexin downregulation. All together, the dynamic labelling of cortical structures we observed strongly supports involvement of calpactin-like annexins in trichocyst docking. Anti-B15 Abs recognized a band of 51 kDa and some of higher values. These Abs selectively labelled the outlines of the cytoproct, the site of spent phagolysosome exocytosis. In conclusion, our data indicate involvement of specific sets of annexins in site-specific positioning and attachment of widely different secretory organelles at the cell surface in Paramecium cells.

Meeting the learning needs of senior citizens and nursing students through a community-based pharmacology experience.

Senior citizens commonly take multiple medications and lack knowledge about proper use and side effects. They often have no access to health professionals to clarify any misconceptions about their medications. Nursing students often learn pharmacology in the classroom with no opportunity to apply the knowledge as it is learned. Within one community, a creative strategy was developed to meet the learning needs of both groups. At a local senior center, clients with drug-knowledge deficits or compliance difficulties were identified. Pharmacology nursing students were then assigned to collect drug histories from these clients. Following analysis of gathered data, students were required to develop individualized drug teaching tools for their assigned client. Through these activities, the learning needs of both groups were met.

Caffeine inhibits Ca2+ uptake by subplasmalemmal calcium stores ('alveolar sacs') isolated from Paramecium cells.

Caffeine inhibits 45Ca2+ sequestration by subplasmalemmal calcium stores ('alveolar sacs') of low thapsigargicin sensitivity which we have isolated from the ciliated protozoan, Paramecium tetraurelia. Inhibition depends on caffeine concentration, with an IC50 of 31.8 mM. According to kinetic evaluation this is compatible with non-competitive inhibition of Ca2+ uptake, rather than with superimposed 45Ca2+ release during sequestration. It remains to be analysed whether this mechanism might be of possible relevance also for Ca2+-mediated activation in vivo in this or in any other secretory system. Such an effect could also operate indirectly, e.g., by Ca2+-release induction via sequestration inhibition. This is the first description of caffeine-mediated inhibition of Ca2+ uptake by calcium stores from a secretory system. Our data are compatible with some observations with sarcoplasmic reticulum from striated muscle fibers.

Strategies for teaching critical thinking in pharmacology.

The author presents six strategies that have been used successfully to teach and assess critical-thinking skills in a pharmacology course. Within the context of pharmacology content, the strategies were designed to develop student abilities in making critical distinctions, critical decision making, analysis of assessment findings, and the anticipation of intervention results and possible side effects/consequences.

A 63 kDa phosphoprotein undergoing rapid dephosphorylation during exocytosis in Paramecium cells shares biochemical characteristics with phosphoglucomutase.

We have enriched phosphoglucomutase (PGM; EC 5.4.2.2) approximately 20-fold from Paramecium tetraurelia cells by combined fractional precipitation with (NH4)2SO4, gel filtration and anion-exchange chromatography yielding two PGM peaks. Several parameters affecting PGM enzymic activity, molecular mass and pI were determined. Phosphorylation studies were done with isolated endogenous protein kinases. Like the 63 kDa phosphoprotein PP63, which is dephosphorylated within 80 ms during synchronous trichocyst exocytosis [Höhne-Zell, Knoll, Riedel-Gras, Hofer and Plattner (1992) Biochem. J. 286, 843-849], PGM has a molecular mass of 63 kDa and forms of identical pI. Since mammalian PGM activity depends on the presence of glucose 1,6-bisphosphate (Glc-1,6-P2) (which is lost during anion-exchange chromatography), we analysed this aspect with Paramecium PGM. In this case PGM activity was shown not to be lost, due to p-nitrophenyl phosphate-detectable phosphatase(s) (which we have separated from PGM), but also due to loss of Glc-1,6-P2. Like PGM from various vertebrate species, PGM activity from Paramecium can be fully re-established by addition of Glc-1,6-P2 at 10 nM, and it is also stimulated by bivalent cations and insensitive to chelating or thiol reagents. The PGM which we have isolated can be phosphorylated by endogenous cyclic-GMP-dependent protein kinase or by endogenous casein kinase. This results in three phosphorylated bands of identical molecular mass and pI values, as we have shown to occur with PP63 after phosphorylation in vivo (forms with pI 6.05, 5.95, 5.85). In ELISA, antibodies raised against PGM from rabbit skeletal muscle were reactive not only with original PGM but also with PGM fractions from Paramecium. Therefore, PGM and PP63 seem to be identical with regard to widely different parameters, i.e. co-elution by chromatography, molecular mass, phosphorylation by the two protein kinases tested, pI values of isoforms, and immuno-binding. Recent claims that PP63 ('parafusin') would not be identical with PGM specifically in Paramecium are critically evaluated. Since some glycolytic enzymes are discussed as being associated with the Ca(2+)-release channel in muscle sarcoplasmic reticulum, and since sub-plasmalemmal Ca2+ stores in Paramecium closely resemble sarcoplasmic reticulum, a possible function of PP63/PGM in exocytosis regulation is discussed, particularly since dephosphorylation strictly parallels exocytosis.

Teaching nursing theory through an election campaign framework.

The author describes a teaching-learning strategy that helped students gain an enhanced appreciation for nursing theories and their application to nursing practice. Students selected, campaigned for, and elected a nursing theorist to serve as Vice President for a Nursing Theory Club. Through the campaign and election process, they were able to select and investigate a nursing theorist and apply the theorists' work to nursing practice. Campaign materials, campaign headquarter displays, and campaign speeches allowed for assessment of many program outcomes including effective written and verbal communication, critical thinking, professionalism, empiric knowledge, and aesthetics.

[Psychopathologic changes in chronic schizophrenic patients following withdrawal of neuroleptics: relation to therapeutic, neuroendocrine and computer tomography variables]. / Psychopathologische Veränderungen bei chronisch schizophrenen Patienten nach Neuroleptikaentzug: Beziehungen zu therapeutischen, neuroendokrinen und computertomographischen Variablen.

The following data were obtained from 36 male chronic schizophrenics (ICD 295.6) of 47 +/- 11 years of age, treated with neuroleptics for the last 16 +/- 6 years: Age, age at first manifestation of disease, duration and dosage scheme of neuroleptic therapy and number of electroconvulsive shock treatments. Blood samples were drawn both under neuroleptic treatment and after a 12-day withdrawal of neuroleptics, for determining cortisol, prolactin, beta-endorphin and noradrenaline. Psychopathology was assessed by standard criteria via BPRS. In 27 patients CT determination was carried out to determine the breadth of the third ventricle and the ventricular brain ratio. Withdrawal of neuroleptics resulted in a marked improvement, whereas 11 patients showed pronounced deterioration of their psychotic symptoms. In respect of the entire group there was a significant improvement of anergy, while disturbances of thinking were significantly enhanced. Serum levels of beta-endorphin and cortisol increased after neuroleptic withdrawal, whereas the levels of prolactin and of noradrenaline dropped. A considerable proportion of the patients showed a distinct extension of the ventricular system, but the CT variables correlated only slight with psychopathological parameters or their changes after neuroleptic withdrawal. The other variables, too, were without clinically relevant prognostic importance compared with the psychopathological changes after neuroleptic withdrawal. These variables were e.g. demography, psychopathology, therapy and neuroendocrinology.

Neuroleptic withdrawal in chronic schizophrenia: CT and endocrine variables relating to psychopathology.

The psychopathology of 36 chronic, schizophrenic patients who had been on maintenance neuroleptics was rated during neuroleptic therapy and after 12 days of neuroleptic withdrawal. At both times, hormonal serum levels were also determined. Moreover, in 27 of these patients, ventricle-brain ratio, maximal width of the third ventricle, and sulcal widening were measured on computed tomography (CT). Neuroleptic withdrawal resulted in individually different psychopathological changes: 7 patients improved, 11 worsened. Within the whole group, thought disorder deteriorated, and anergia improved. Levels of cortisol and beta-endorphin increased; those of prolactin and norepinephrine decreased. The majority of patients showed ventricular enlargement, which was marginally related to reduced thought disorder. CT and endocrine variables were slightly related to psychopathology or psychopathological effects of neuroleptic withdrawal.