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Angiotensin II elicits cytosolic Ca2+ signal that is transferred into the mitochondria. Previously we found in H295R cells that this signal transfer is enhanced by both the inhibition of p38 MAPK and a novel isoform of PKC [G. Szanda, P. Koncz, A. Rajki, A. Spät, Participation of p38 MAPK and a novel-type protein kinase C in the control of mitochondrial Ca2+ uptake, Cell Calcium 43 (2008) 250-259]. Now we report that simultaneous activation of these protein kinases (by TNFalpha and PMA+an inhibitor of the conventional PKC isoforms, respectively) attenuates the transfer of cytosolic Ca2+ signal, elicited by depolarisation or store-operated Ca2+ influx, into the mitochondria. The Ca2+ uptake enhancing effect of the p38 MAPK inhibitor SB202190 is due to the inhibition of p38 MAPK and not to a direct mitochondrial action. Protein kinases reduce mitochondrial [Ca2+] by inhibiting the uptake mechanism. The threshold of mitochondrial Ca2+ uptake may depend on the activity of p38 MAPK. The silencing of protein kinase D (PKD) also results in enhanced transfer of Ca2+ signal from the cytosol into the mitochondria. Our data indicate that Ca2+ mobilising agonists, through the simultaneous activation of p38 MAPK, a novel PKC isoform and PKD, exert a negative feed-forward action on mitochondrial Ca2+ uptake, thus reducing the risk of Ca2+ overload.

Cytosolic Ca2+ signals are followed by mitochondrial Ca2+ uptake, which, in turn, modifies several biological processes. Mg2+ is known to inhibit Ca2+ uptake by isolated mitochondria, but its significance in intact cells has not been elucidated. In HEK293T cells, activation of purinergic receptors with extracellular ATP caused cytosolic Ca2+ signals associated with parallel changes in cytosolic [Mg2+]. Neither signals were affected by omitting bivalent cations from the extracellular medium. The effect of store-operated Ca2+ influx on cytosolic Mg2+ concentration ([Mg2+]c) was negligible. Uncaged Ca2+ displaced Mg2+ from cytosolic binding sites, but for an equivalent Ca2+ signal, the change in [Mg2+] was significantly smaller than that measured after adding extracellular ATP. Inositol 1,4,5-trisphosphate mobilized Ca2+ and Mg2+ from internal stores in permeabilized cells. The increase of [Mg2+] in the range that occurred in ATP-stimulated cells inhibited mitochondrial Ca2+ uptake in permeabilized cells without affecting mitochondrial Ca2+ efflux. Therefore, the Mg2+ signal generated by Ca2+ mobilizing agonists may attenuate mitochondrial Ca2+ uptake.

Clinical practice requires knowledge of health professionals for the application of problem solving of different tasks in various clinical settings. Therefore, a set of reliable measurement tools is required to assess various components of the professional knowledge including factual knowledge, skills and competence as outcomes of nursing education and training. Traditional questionnaires and test methods do not allow the measurement of these characteristics properly, as these tools typically provide overall scores without relevant insight into areas in which nurses perform well, and those where their knowledge and/or skills should be improved substantially.The aim of this nationwide survey conducted among nurses between November 2014 and February 2015 was to determine how the teaching/training objectives have been achieved in the nurses' transfusion practice. The authors attempted to exceed the capabilities of classical test theory and acquire a detailed picture about what the nurses know about transfusion therapy and how they are involved and behave in routine clinical practice.The knowledge and skills of 657 participants were assessed using a validated instrument consisting of a set of questions covering every aspects of transfusion therapy. The answers to these items were evaluated on a binary (good or bad) scale. Recorded answers of the participants were analysed using hierarchical cluster analysis and item response theory tools such as the one-parametric Rasch model suitable for dichotomous data. Data analysis was performed with the SPSS program and the ltm module of the R statistical program.The paper presents the distribution of correct and incorrect answers to various questions about transfusion therapy along with the corresponding logit values and odds ratios, respectively. The characteristic curves of each item were determined on the basis of the number of correct answers that have been recorded. These curves highlight which questions were answered easily and which items were found harder to answer by the nurses who participated in the survey. In addition to the separate analysis of individual questions, a set of response patterns is also presented which shows how frequently the nurses responded correctly to different combinations (sub-sets) of questions. On the whole, nurses exhibited medium level performance in terms of knowledge and skills required for efficient and safe transfusion practice.Objective and reliable measurement of the level of acquired knowledge is a key requirement in nursing education. This paper, which demonstrates the use of cluster analysis and item response theory for the assessment of transfusion-related knowledge of nurses, focuses on this issue for the first time in nursing research. The results of this survey have revealed substantial limitations and deficiencies both in knowledge and skills of nurses which need to be addressed by training in order to improve the efficiency and safety of transfusion therapy.

International literature data clearly show that the role of nurses in transfusion therapy, which needs professional knowledge and skills, continues to increase.The aim of the authors was to perform a national-level mapping of the practice of transfusion therapy including transfusion therapy-related knowledge of nurses and their knowledge about their competence. Further aim was to identify similarities and differences as referred to the requirements of the transfusion regulations (2008), which was valid at the time of the survey in Hungary.The nationwide survey took place between November 19, 2014 and February 20, 2015. Data were collected using paper-form and web-based anonymous, self-administered questionnaire. 657 nurses who were involved in transfusion therapy with changing regularity in adult in-patient wards took part in the survey. Data were processed using descriptive statistical methods.It was found that the transfusion therapy related knowledge of nurses was very incomplete (50.72%), and only about half of the nurses (52.3%) systematized the nursing functions correctly. Significant differences were also found in the practice of transfusion therapy. The vast majority of the institutions had their own protocol, but responsible nurses assigned for transfusion care were present only in 23.9% of the wards.The authors conclude that the identified problems can be resolved by improving professional responsibility of nurses, increasing supervision by nursing leaders and appropriate transfusiology training and regular retrainings.

Glomerulosa cells secrete aldosterone in response to agonists coupled to Ca(2+) increases such as angiotensin II and corticotrophin, coupled to a cAMP dependent pathway. A recently recognized interaction between Ca(2+) and cAMP is the Ca(2+)-induced cAMP formation in the mitochondrial matrix. Here we describe that soluble adenylyl cyclase (sAC) is expressed in H295R adrenocortical cells. Mitochondrial cAMP formation, monitored with a mitochondria-targeted fluorescent sensor (4mtH30), is enhanced by HCO3(-) and the Ca(2+) mobilizing agonist angiotensin II. The effect of angiotensin II is inhibited by 2-OHE, an inhibitor of sAC, and by RNA interference of sAC, but enhanced by an inhibitor of phosphodiesterase PDE2A. Heterologous expression of the Ca(2+) binding protein S100G within the mitochondrial matrix attenuates angiotensin II-induced mitochondrial cAMP formation. Inhibition and knockdown of sAC significantly reduce angiotensin II-induced aldosterone production. These data provide the first evidence for a cell-specific functional role of mitochondrial cAMP.

All people involved in transfusion therapy have professional, legal and ethical responsibility for their own actions.The aims of the authors were to evaluate competencies, knowledge and practice of nurses about transfusion therapy.Descriptive statistical approach using observation, questionnaire survey and interview was applied. Reliability and validity of the self-made questionnaire were examined with statistical methods. Variance, standard deviation, Cronbach's alpha and Pearson correlations were calculated.Transfusiology-related knowledge of nurses regarding current transfusion regulations was almost 90%, and 56.2% of nurses systematized the nursing functions correctly. Significant differences were found among different institutions in transfusion therapy, transfusion practice and the use of relevant documentation. 75-77% of the institutions examined had their own protocol, and assigned transfusion care nurses worked in more than 20% of the wards.The authors propose a better supervision by leaders in nursing aspects of transfusion therapy, and an increased professional responsibility.Bevezetés: A transzfúziós terápiában részt vevő valamennyi személy szakmai, jogi és etikai felelősséggel tartozik saját tevékenységéért. A kompetenciák megfogalmazásához elengedhetetlen az ápolás funkcióinak meghatározása. Célkitűzés: A szerzők célul tűzték ki a transzfúziós terápia gyakorlatának, valamint az ápolók transzfúziós terápiával kapcsolatos ismereteinek feltérképezését. Módszer: A vizsgálathoz leíró statisztikai megközelítésben a megfigyelés, a kérdőíves felmérés és az interjú módszerét választották. A saját készítésű kérdőív megbízhatóságát és érvényességét matematikai statisztikai módszerekkel vizsgálták, amelynek során variancia-, standarddeviáció-, valamint Cronbach-alfa-számítást és Pearson-korreláció-meghatározást végeztek. Eredmények: Az ápolók transzfuziológiai ismeretei az érvényes transzfúziós szabályzat tekintetében megközelítették a 90%-ot, az ápolási funkciókat az ápolók 56,2%-a rendszerezte helyesen. Az intézményeknél jelentős különbséget találtak a transzfúziós terápia gyakorlatára, az idevonatkozó dokumentációk alkalmazására vonatkozóan. Az intézmények 75–77%-a rendelkezik saját protokollal, és a betegosztályok ötödén dolgozik megbízott transzfúziós felelős ápoló. Következtetések: A szerzők a megoldást az ápolás vezetőinek a transzfúziós terápia ápolási vonatkozásainak nagyobb mértékű felügyeletében, a kollégák szakmai felelősségének növelésében látják. Orv. Hetil., 2015, 156(9), 366–378.

INTRODUCTION AND OBJECTIVES: Sacral nerve stimulation (SNS) is FDA-approved for refractory lower urinary tract and bowel dysfunction. Improved surgical technique and stimulator programming have reduced complications and extended device life. Troubleshooting methods and case series of complications have been documented, but shed little insight into physical lead failures. This study described patterns and rates of physical lead failures with abnormal impedances in a multi-surgeon, single-center experience to formulate potential recommendations for programming, implantation, and improvements in lead manufacturing. METHODS: All cases between September 2007 and 2011 with abnormal lead impedances were identified. A manual review of the electronic medical record was conducted to collect patient demographics, comorbidities, procedural details, program settings, and device interrogation results. RESULTS: A total of 24 hardware-related lead failures were identified by abnormal impedance readings, of which 92% (22/24) were open-circuit (impedance 4000 Ohms) and 8% (2/24) were shortcircuit (impedance 400 Ohms) failures. Coincidentally, all patients had a primary indication of refractory urgency-frequency and underwent a staged, unilateral implantation. Median age at implant was 48[IQR: 42-64] years and BMI was 28.7[24.1-33.5] kg/m2. Failure was noted at 84[26-152] weeks after implant, with 1 patient having a second failure 74 weeks after re-implantation. Amplitude after permanent implantation was 1.7[0.9-2.2] V and 2[1.1-4.1] V at the time of failure. A single electrode accounted for 45.8% of failures, while 54.2% had multi-electrode failures. Electrode 0 (most distal) and 3 (most proximal) each accounted for 33% of failures, while electrodes 1 and 2 accounted for 17.8% and 15.6%, respectively. CONCLUSIONS: Electrode 0 (most distal) and 3 (most proximal) appear to have the highest risk of open-circuit and short-circuit failure in this retrospective series. Moving forward, optimization of the other electrode (i.e. 1 and 2) responses during implantation and reprogramming around electrodes 0 and 3, if damaged, may help minimize lead-related therapeutic failures necessitating surgical revision.

Two experiments were performed to investigate the role of extra-legal factors in a simulated product liability trial. In cases where the factual evidence was identical, subjects' liability judgments varied as a function of the case-specific factor of the alleged source of the plaintiffs injury. In deciding cases differently depending on the alleged cause, subjects relied on intuitions about what injury sources are more or less likely to cause a certain kind of injury. Juror-specific factors also influenced subjects' verdicts. There was no difference between students and non-students, but race and SEC—factors that are often correlated with student status—did affect subjects' verdicts. Low-SES and minority subjects were more likely to find the defendant liable than high-SES and white subjects. The results are considered in terms of general decision-making processes, and the implications for jury selection and mock jury research are discussed.

Angiotensin II elicits cytosolic and mitochondrial Ca2+ signal in H295R adrenocortical cells. We found that Ca2+ uptake rate and peak values in small mitochondrial regions both depend on the colocalization of these mitochondrial regions with GFP-marked endoplasmic reticular (ER) vesicles. The dependence of the Ca2+ response on this colocalization is abolished by SB202190 and PD169316, inhibitors of p38 MAPK, as well as by transfection with siRNA against p38 MAPK mRNA. The same manoeuvres result in an increased ratio of global mitochondrial to global cytosolic Ca2+ response, indicating that inhibition of p38 MAPK is followed by enhanced mitochondrial Ca2+ uptake. alpha-Toxin and TNFalpha, agents which similarly to angiotensin II increase the phosphorylation of p38, failed to affect mitochondrial Ca2+ uptake, indicating that activation of p38 MAPK is necessary but not sufficient for the inhibition of Ca2+ uptake. Bisindolylmaleimide, an inhibitor of the conventional and novel-type protein kinase C isoforms also evokes enhanced mitochondrial Ca2+ uptake, whereas Gö6976 that inhibits the conventional isoforms only failed to exert any effect. These data show that angiotensin II attenuates Ca2+ uptake predominantly into mitochondria that do not colocalize with ER, by a mechanism involving p38 MAPK and a novel-type PKC.

The acute effects of ultraviolet light, the superoxide-generating xanthine-xanthine oxidase system and H(2)O(2) to on calcium signaling and mitochondrial pyridine nucleotide metabolism were investigated in rat glomerulosa cells. UV light induced the formation of superoxide, that, similar to exogenously applied superoxide and H(2)O(2), decreased the level of mitochondrial NAD(P)H. Free radical scavengers antagonized this effect of UV light. Extracellularly generated superoxide elicited Ca(2+) transients and inhibited angiotensin II-induced cytoplasmic Ca(2+) signaling. Low intensity UV light did not affect basal [Ca(2+)] and failed to influence Ca(2+) signaling induced by depolarization or store depletion. UV light of the same low power reduced both cytoplasmic and mitochondrial Ca(2+) signals induced by angiotensin II. The lack of UV effect on inositol phosphate formation indicates that the inhibition of cytoplasmic Ca(2+) signaling is due to reduced Ca(2+) release from InsP(3)-sensitive stores. Decreased mitochondrial Ca(2+) uptake may be attributed to UV-induced perturbation of the perimitochondrial microdomain.

Rapid detection and quantitative assessment of specific microbial species in environmental samples is desirable for monitoring changes in ecosystems and for tracking natural or introduced microbial species during bioremediation of contaminated sites. In the interests of developing rapid tests for hydrocarbon-degrading bacteria, species-specific PCR primer sets have been developed for Pseudomonas aeruginosa, Stentrophomonas (Xanthomonas) maltophilia, and Serratia marsescens. Highly variable regions of the 16S rRNA gene were used to design these primer sets. The amplification products of these primer sets have been verified and validated with hemi-nested PCR and with ligase chain reaction (LCR) techniques, and have been applied to the analyses of environmental water samples. These species-specific primer sets were also chosen to amplify in conjunction with a universal set of PCR primers chosen from highly conserved neighboring sequences in the same gene. These multiplex or competitive PCR procedures enable testing with an internal marker and/or the quantitative estimation of the relative proportion of the microbial community that any one of these species occupies. In addition, this universal PCR primer set amplified the same size amplicon from a wide spectrum of procaryotic and eucaryotic organisms and may have potential in earth biota analyses.

Zinc, cadmium and bismuth have been electrodeposited on copper from tetramethylurea, a new organic electrolyte. The nature of the deposits so obtained was good. A more quantitative treatment of the electro-deposition of these metals from this solvent is suggested.

Voltage-gated proton current (I(Hv)) has been characterized in several cell types, but the majority of the data was collected in phagocytes, especially in human granulocytes. The prevailing view about the role of I(Hv) in phagocytes is that it is an essential supporter of the intense and sustained activity of Nox2 (the core enzyme of the phagocyte NADPH oxidase complex) during respiratory burst. Recently H(v)1, a voltage-gated proton channel, was cloned, and leukocytes from H(v)1 knockout mice display impaired respiratory burst. On the other hand, hardly anything is known about H(v)1 in human granulocytes. Using qPCR and a self made antibody, we detected a significant amount of H(v)1 in human eosinophil and neutrophil granulocytes and in PLB-985 leukemia cells. Using different crosslinking agents and detergents in reducing and non-reducing PAGE, significant expression of H(v)1 homodimers, but not that of higher-order multimers, could be detected in granulocytes. Results of subcellular fractionation and confocal imaging indicate that H(v)1 is resident in both plasmalemmal and granular membrane compartments of resting neutrophils. Furthermore, it is also demonstrated that H(v)1 accumulates in phagosome wall during zymosan engulfment together with, but independently of Nox2. During granulocytic differentiation early and parallel upregulation of H(v)1 and Nox2 expression was observed in PLB-985 cells. The upregulation of H(v)1 or Nox2 expression did not require the normal expression of the other molecule. Using RNA interference, we obtained strong correlation between H(v)1 expression and I(Hv) density in PLB-985 cells. It is also demonstrated that a massive reduction in H(v)1 expression can limit the Nox2 mediated superoxide production of PLB-985 granulocytes. In summary, beside monomers native H(v)1 forms stable proton channel dimer in resting and activated human granulocytes. The expression pattern of H(v)1 in granulocytes is optimized to support intense NADPH oxidase activity.