A heterotypic assembly mechanism regulates CHIP E3 ligase activity.

CHIP (C-terminus of Hsc70-interacting protein) and its worm ortholog CHN-1 are E3 ubiquitin ligases that link the chaperone system with the ubiquitin-proteasome system (UPS). CHN-1 can cooperate with UFD-2, another E3 ligase, to accelerate ubiquitin chain formation; however, the basis for the high processivity of this E3s set has remained obscure. Here, we studied the molecular mechanism and function of the CHN-1-UFD-2 complex in Caenorhabditis elegans. Our data show that UFD-2 binding promotes the cooperation between CHN-1 and ubiquitin-conjugating E2 enzymes by stabilizing the CHN-1 U-box dimer. However, HSP70/HSP-1 chaperone outcompetes UFD-2 for CHN-1 binding, thereby promoting a shift to the autoinhibited CHN-1 state by acting on a conserved residue in its U-box domain. The interaction with UFD-2 enables CHN-1 to efficiently ubiquitylate and regulate S-adenosylhomocysteinase (AHCY-1), a key enzyme in the S-adenosylmethionine (SAM) regeneration cycle, which is essential for SAM-dependent methylation. Our results define the molecular mechanism underlying the synergistic cooperation of CHN-1 and UFD-2 in substrate ubiquitylation.

HaDeX: an R package and web-server for analysis of data from hydrogen-deuterium exchange mass spectrometry experiments.

MOTIVATION: Hydrogen-deuterium mass spectrometry (HDX-MS) is a rapidly developing technique for monitoring dynamics and interactions of proteins. The development of new devices has to be followed with new software suites addressing emerging standards in data analysis. RESULTS: We propose HaDeX, a novel tool for processing, analysis and visualization of HDX-MS experiments. HaDeX supports a reproducible analytical process, including data exploration, quality control and generation of publication-quality figures. AVAILABILITY AND IMPLEMENTATION: HaDeX is available primarily as a web-server (http://mslab-ibb.pl/shiny/HaDeX/), but its all functionalities are also accessible as the R package (https://CRAN.R-project.org/package=HaDeX) and standalone software (https://sourceforge.net/projects/HaDeX/). SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Left ventricular diastolic dysfunction and diseases severity contribute to impaired exercise capacity in systemic lupus erythematosus.

OBJECTIVES: Patients with systemic lupus erythematosus (SLE) have a higher risk of myocardial involvement, which can result in ventricular dysfunction. The aim of our study was to estimate potential relationship between exercise capacity assessed by six minute walk test (6MWT) and echocardiographic parameters of left and right ventricular function in SLE patients. METHODS: We prospectively studied 66 SLE patients (57 F, age 44 (20-75) years) and 27 age matched healthy subjects. In addition to routine evaluation, 6MWT and transthoracic echocardiography including LV diastolic dysfunction parameters (E/A, E/É) were performed. RESULTS: While E/A was similar in both groups, E/E' was higher in patients with SLE than in controls, 7.5 (4-22) vs 6.8 (1.6-9.4), p = 0.018. The mean 6MWT distance was significantly shorter in SLE (561.6 ± 150.7 vs 682.6 ± 98.1 m, p < 0.002). Among SLE patients only 53 (80.3%) were capable to walk at least 450 m, while in controls 27 (100%) (p = 0.013). We observed significant correlations between 6MWT distance and SLICC/ACR-DI (rho=-0.44, p < 0.001), E/A (rho = 0.30, p = 0.004), E/E' (rho=-0.36, p < 0.001) in SLE patients. Univariable logistic regression models revealed that SLICC/ACR-DI, E/E', tricuspid regurgitant peak gradient (TRPG), and right ventricular systolic pressure (RVSP) were associated with 6MWT distance lower than < 450 m. ROC curves shown high predictive value of E/E' ratio, TRPG, RVSP in the prediction for 6MWT distance < 450 m. CONCLUSION: Impaired exercise tolerance seems to result mainly from the severity of SLE and LV diastolic dysfunction.

The Role of Nrf2 Transcription Factor and Sp1-Nrf2 Protein Complex in Glutamine Transporter SN1 Regulation in Mouse Cortical Astrocytes Exposed to Ammonia.

Ammonia toxicity in the brain primarily affects astrocytes via a mechanism in which oxidative stress (OS), is coupled to the imbalance between glutamatergic and GABAergic transmission. Ammonia also downregulates the astrocytic N system transporter SN1 that controls glutamine supply from astrocytes to neurons for the replenishment of both neurotransmitters. Here, we tested the hypothesis that activation of Nrf2 is the process that links ammonia-induced OS formation in astrocytes to downregulation and inactivation of SN1 and that it may involve the formation of a complex between Nrf2 and Sp1. Treatment of cultured cortical mouse astrocytes with ammonia (5 mM NH4Cl for 24 h) evoked Nrf2 nuclear translocation, increased its activity in a p38 MAPK pathway-dependent manner, and enhanced Nrf2 binding to Slc38a3 promoter. Nrf2 silencing increased SN1 mRNA and protein level without influencing astrocytic [3H]glutamine transport. Ammonia decreased SN1 expression in Nrf2 siRNA treated astrocytes and reduced [3H]glutamine uptake. In addition, while Nrf2 formed a complex with Sp1 in ammonia-treated astrocytes less efficiently than in control cells, treatment of astrocytes with hybrid-mode inactivated Sp1-Nrf2 complex (Nrf2 silencing + pharmacological inhibition of Sp1) did not affect SN1 protein level in ammonia-treated astrocytes. In summary, the results document that SN1 transporter dysregulation by ammonia in astrocytes involves activation of Nrf2 but does not require the formation of the Sp1-Nrf2 complex.

Psychometric properties of the Polish version of two screening tests for gambling disorders: the Problem Gambling Severity Index and Lie/Bet Questionnaire.

To date, no screening tests for gambling disorders have been adapted and validated in Central and Eastern Europe. The aim of this study is to adapt the Problem Gambling Severity Index (PGSI) and Lie/Bet questionnaire (Lie/Bet) and assess their psychometric properties once translated for use with the Polish population. A mixed sample (N = 300) was drawn from venues, social media, snowballing and treatment centers. PGSI had a higher coefficient of predictive power than Lie/Bet. However, differences between validated tests are not significant. Validation of screening tests of gambling disorders showed the necessity for verification of the scale of interpretation of results when conducting tests in Poland, changing cutoff values. The PGSI and Lie/Bet tests are short and easy to apply, they can be implemented in various types of institutions: for screening patients in primary health care facilities and for identifying comorbid gambling disorders in alcohol- and drug-dependence treatment facilities, and in social welfare centers.

Synergistic interaction of diclofenac and its metabolites with selected antibiotics and amygdalin in wastewaters.

Diclofenac (DCF), a non-steroidal anti-inflammatory drug (NSAID) belongs to one of the most frequently detected pharmaceutical residues in the environment. Little is known on the interactions of DCF as well as its major biodegradation metabolites 4'-OHDCF and 5-OHDCF with chemical compounds found in wastewater, including antibiotics such as ampicillin and kanamycin. In the present work we examined the potential interactions between DCF, its metabolites 4'-OHDCF and 5-OHDCF and ampicyllin and kanamycin. We also measured the effect of the mixture of DCF with natural compound - amygdalin. We evaluated the following parameters: E. coli K-12 cells viability, growth inhibition of E. coli K-12 culture, genotoxicity, oxidative stress parameters: sodA promoter induction and ROS generation. The reactivity of E. coli SM recA:luxCDABE biosensor strain in wastewaters matrices contaminated with DCF and kanamycin was also monitored. Obtained results indicated that used antibiotics (ampicyllin, kanamycin) enhanced the toxic effect of DCF used individually and in the mixtures with its metabolites 4'-OHDCF and 5-OHDCF toward E. coli. Similar effect was also obtained in genotoxicity assay. The oxidative stress assays revealed that the highest level of ROS generation and sodA promoter induction were obtained also for the mixtures of DCF, its metabolites with antibiotics. It was also showed that amygdalin influenced the activity of DCF and its biodegradation metabolites. The strongest luminescence response of E. coli SM biosensor strain with recA:luxCDABE genetic construct in filtered treated wastewaters, comparable to control sample was noticed. Obtained results showed that DCF and its biodegradation metabolites 4'-OHDCF and 5-OHDCF can interact with tested antibiotics and compounds of natural origin, i.e. amygdalin to form mixtures showing stronger antimicrobial activity against E. coli than parent chemicals. Moreover the assays in wastewater matrices revealed that E. coli SM recA:luxCDABE biosensor strains is a good tool for bacteria monitoring in wastewater environments.

The study of biological activity of transformation products of diclofenac and its interaction with chlorogenic acid.

In the present work we compared the biological activity of DCF, 4'-OHDCF and 5-OHDCF as molecules of most biodegradation pathways of DCF and selected transformation products (2-hydroxyphenylacetic acid; 2,5-dihydroxyphenylacetic acid and 2,6-dichloroaniline) which are produced during AOPs, such as ozonation and UV/H2O2. We also examined the interaction of DCF with chlorogenic acid (CGA). CGA is commonly used in human diet and entering the environment along with waste mainly from the processing and brewing of coffee and it can be toxic for microorganisms included in activated sludge. In the present experiment the evaluation of following parameters was performed: E. coli K-12 cells viability, growth inhibition of E. coli K-12 culture, LC50 and mortality of Chironomus aprilinus, genotoxicity, sodA promoter induction and ROS generation. In addition the reactivity of E. coli SM recA:luxCDABE biosensor strain in wastewater matrices was measured. The results showed the influence of DCF, 4'-OHDCF and 5-OHDCF on E. coli K-12 cells viability and bacteria growth, comparable to AOPs by-products. The highest toxicity was observed for selected, tested AOPs by-products, in comparison to the DCF, 4'-OHDCF and 5-OHDCF. Genotoxicity assay indicated that 2,6-dichloroaniline (AOPs by-product) had the highest toxic effect. The oxidative stress assays revealed that the highest level of ROS generation and sodA promoter induction were obtained for DCF, 4'-OHDCF and 5-OHDCF, compared to other tested compounds. We have also found that there is an interaction between chlorogenic acid and DCF, which resulted in increased toxicity of the mixture of the both compounds to E. coli K-12, comparable to parent chemicals. The strongest response of E. coli SM biosensor strain with recA:luxCDABE genetic construct in filtered treated wastewaters, comparable to control sample was noticed. It indicates, that E. coli SM recA:luxCDABE biosensor strains is a good tool for bacteria monitoring in wastewater environment. Due to toxicity and biological activity of tested DCF transformation products, there is a need to use additional wastewater treatment systems for wastewater contaminated with pharmaceutical residues.

Silencing of Transcription Factor Sp1 Promotes SN1 Transporter Regulation by Ammonia in Mouse Cortical Astrocytes.

The involvement of the astrocytic SN1 (SNAT3) transporter in ammonia-induced l-glutamine retention was recently documented in mouse-cultured astrocytes. Here we investigated the involvement of specificity protein 1 (Sp1) transcription factor in SN1 regulation in ammonium chloride ("ammonia")-treated astrocytes. Sp1 expression and its cellular localization were determined using real-time qPCR, Western blot, and confocal microscopy. Sp1 binding to Snat3 promoter was analyzed by chromatin immunoprecipitation. The role of Sp1 in SN1 expression and SN1-mediated [³H]glutamine uptake in ammonia-treated astrocytes was verified using siRNA and mithramycin A. The involvement of protein kinase C (PKC) isoforms in Sp1 level/phosphorylation status was verified using siRNA technology. Sp1 translocation to the nuclei and its enhanced binding to the Snat3 promoter, along with Sp1 dependence of system N-mediated [³H]glutamine uptake, were observed in astrocytes upon ammonia exposure. Ammonia decreased the level of phosphorylated Sp1, and the effect was reinforced by long-term incubation with PKC modulator, phorbol 12-myristate 13-acetate, which is a treatment likely to dephosphorylate Sp1. Furthermore, silencing of the PKCδ isoform appears to enhance the ammonia effect on the Sp1 level. Collectively, the results demonstrate the regulatory role of Sp1 in regulation of SN1 expression and activity in ammonia-treated astrocytes and implicate altered Sp1 phosphorylation status in this capacity.

Correction to: Psychiatric comorbidity and intimate partner violence among women who inject drugs in Europe: a cross-sectional study.

The original version of this article unfortunately missed the Acknowledgment.

Psychiatric comorbidity and intimate partner violence among women who inject drugs in Europe: a cross-sectional study.

Women who inject drugs (WWID) are an especially vulnerable group of drug users. This study determined the prevalence of psychiatric comorbidity and intimate partrner violence (IPV), and factors associated with psychiatric comorbidity among WWID recruited from drug treatment services (67%) and harm reduction services in five European regions in Austria, Catalonia, Italy, Poland, and Scotland. Psychiatric comorbidity was assessed among 226 WWID using the Dual Diagnosis Screening Instrument. IPV was assessed using the Composite Abuse Scale and injecting and sexual risk behaviors were assessed using a battery of questionnaires adapted and developed for the study. Eighty-seven percent met criteria for at least one lifetime psychiatric disorder. The most common disorders were depression (76%), panic (54%), and post-traumatic stress (52%). WWID recruited in drug treatment services were almost three times as likely (OR 2.90 95% CI 1.30-6.43; p = 0.007) to meet criteria for a lifetime psychiatric disorder than those recruited from harm reduction services, specifically dysthymia (OR 5.32 95% CI 2.27-12.48; p = 0.000) and post-traumatic stress disorder (OR 1.83 95% CI 1.02-3.27; p = 0.040). WWID who reported sharing needles and syringes were almost three times as likely to meet criteria for lifetime psychiatric comorbidity than those who did not (OR 2.65 95% CI 1.07-6.56). Compared to WWID who had not experienced IPV, victims (70%) were almost two times more likely to meet criteria for post-traumatic stress disorder (OR 1.95 95% CI 1.10-3.48). Psychiatric comorbidity and IPV among WWID are common. Drug treatment and harm reduction services should address psychiatric comorbidity and IPV to improve treatment outcomes.

Barriers in Access to the Treatment for People with Gambling Disorders. Are They Different from Those Experienced by People with Alcohol and/or Drug Dependence?

A prevalence of gambling disorders is diversified depending on the region of the world. Almost three quarters of pathological gamblers had never sought a professional treatment as well as an assistance in self-help groups. Reasons why they do not initiate a treatment are complex. The aim of the article is to compare barriers to the treatment for people with gambling disorders found in presented study and barriers to alcohol and drug treatment identified in the available literature. The semi structured interviews were applied and conducted with people with gambling disorders, social workers, therapists employed in the addiction treatment facilities, General Practitioners and psychiatrists. Selection of the respondents was based on purposive sampling. In total, 90 interviews were completed. Respondents identified individual barriers as well as structural ones. Individual barriers include internal resistance and a fear of the treatment. In turn structural barriers apply to the organization of the therapy, infrastructure, personnel, and the therapeutic program. A comparison of barriers experienced by people with gambling disorders and substance use disorders showed that they are largely similar, but people with gambling disorders also experience specific barriers. Empirical studies focused specifically on treatment needs of people experiencing gambling disorders may improve an offer of help for them. More adequate treatment options could contribute to the increasing in the number of people who start the treatment. It can result in improving their quality of life and may have positive impact on public health.

System N transporters are critical for glutamine release and modulate metabolic fluxes of glucose and acetate in cultured cortical astrocytes: changes induced by ammonia.

Glutamine (Gln) is synthesized in astrocytes from glutamate (Glu) and ammonia, whereupon it can be released to be transferred to neurons. This study evaluated the as yet not definitely established role of the astrocytic Gln transporters SN1 and SN2 (Slc38a3 and Slc38a5 respectively) in Gln release and metabolic fluxes of glucose and acetate, the canonical precursors of Glu. Cultured neocortical astrocytes were grown in the absence or presence of ammonia (5 mM NH4 Cl, 24 h), which deregulates astrocytic metabolism in hyperammonemic encephalopathies. HPLC analyses of cell extracts of SN1/SN2 siRNA-treated (SN1/SN2-) astrocytes revealed a ~ 3.5-fold increase in Gln content and doubling of glutathione, aspartate, alanine and glutamate contents, as compared to SN1/SN2+ astrocytes. Uptake and efflux of preloaded [(3) H]Gln was likewise significantly decreased in SN1/SN2- astrocytes. The atom percent excess (13) C values (given as M + 1) for alanine, aspartate and glutamate were decreased when the SN1/SN2- cells were incubated with [1-(13) C] glucose, while Gln consumption was not changed. No difference was seen in M + 1 values in SN1/SN2- cells incubated with [2-(13) C] acetate, which were not treated with ammonia. In SN1/SN2- astrocytes, the increase in Gln content and the decrease in radiolabeled Gln release upon exposure to ammonia were found abrogated, and glutamate labeling from [2-(13) C]acetate was decreased as compared to SN1/SN2+ astrocytes. The results underscore a profound role of SN1 and/or SN2 in Gln release from astrocytes under physiological conditions, but less so in ammonia-overexposed astrocytes, and appear to manifest dependence of astrocytic glucose metabolism to Glu/Gln on unimpaired SN1/SN2- mediated Gln release from astrocytes. The astrocytic N system transporters SN1 and SN2 show preponderance to mediate glutamine (Gln) efflux. Under hyperammonemic conditions, accumulation of Gln, a direct product of ammonia detoxification, may deregulate astrocytic metabolism and seems to be responsible for astrocytic swelling. This study evaluated not definitely established role of SN1 and SN2 in Gln release and metabolic fluxes of radiolabeled glucose and acetate. Simultaneous silencing of SN1/SN2 transporters increase Gln, glutathione, aspartate, alanine and glutamate contents (Panel B; marked in red) as compare to non-silenced astrocytes (Panel A). The atom percent excess (13) C values (given as M + 1) for alanine, aspartate and glutamate were decreased when the cells with silenced transporters were incubated with [1-(13) C]glucose, whereas no difference was seen in M + 1 values when those cells were incubated with [2-(13) C]acetate. Ammonia abrogated the increase in Gln content and decrease in radiolabeled Gln release in astrocytes with silenced transporters, but caused a decrease in glutamate labeling from [2-(13) C]acetate.

Spuriously high androstendione concentrations due to assay interference as a cause of diagnostic conundrum in women with oligomenorrhoea.

UNLABELLED: Polycystic ovary syndrome (PCOS) is a diagnosis of exclusion. We present two cases of women with oligomenorrhoea and high concentration of androstendione, suggestive of possible androgen-secreting tumour; caused by assay interference. The first patient, investigated for oligomenorrhoea, had no significant hirsutism or acne. Androstendione concentration was above 10.0 ng/ml (rr: 0.3-3.3 ng/ml). In order to rule out possible androgen-secreting tumour or hypercortisolaemia we performed 48-hour low dose dexamethasone suppression test (LDDST). This failed to demonstrate adequate suppression of androstendione (6.05 ng/ml and 9.32 ng/ml after the first and the second day respectively). Pelvic ultrasound examination showed polycystic ovaries, while abdominal CTscan failed to show any ovarian or adrenal lesion. Despite such high androstendione concentrations, urinary steroid profile (gas chromatography/mass spectrometry method) yielded normal results. Hence a possibility of androstendione assay interference was raised. The second patient was also admitted for investigations of oligomenorrhoea. Clinical examination was unremarkable. There was a high concentration of testosterone 0.78 ng/ml (rr. 0.084-0.481 ng/ml) and androstendione above 10.0 ng/ml (rr: 0.3-3.3 ng/ml). LDDST failed to demonstrate any suppression of androstendione, while recalculated concentrations of androstendione after serial dilutions were markedly lower in comparison to initial values. Therefore, such high androstendione concentrations (i.e. above the upper limit of the assay) must have resulted from assay interference. In both cases a final diagnosis of PCOS was established. CONCLUSIONS: In the absence of clinical features, contrasting with unusually high androgen levels, a possibility of androgen assay interference should be considered in differential diagnosis of hyperandrogenism or PCOS.

The value of Pa(CO2) in relation to outcome in congenital diaphragmatic hernia.

BACKGROUND: Postnatal assessment of disease severity is critical for analysis of mortality rates and development of future interventions in congenital diaphragmatic hernia (CDH). OBJECTIVE: The objective of this study was to stratify the risk of mortality based on arterial Paco 2. METHODS: Retrospective analysis of infants (n = 133) with CDH admitted to a regional extracorporeal membrane oxygenation (ECMO) center in two different periods: period I (1987-1996; n = 46) and period II (2002-2010; n = 87). RESULTS: The mortality rate (37%) was similar in both periods (p = 0.98). Paco 2 < 60 mm Hg in the first arterial blood gas (ABG) was an independent predictor of survival in both periods (p = 0.03). The predicted survival rate was 84% if initial Paco 2 was < 55 mm Hg. For infants with initial Paco 2 > 55 mm Hg treated with ECMO (n = 83), the predicted survival rate was 11% if the Paco 2 was > 88 mm Hg before the initiation of ECMO. CONCLUSION: Paco 2, a surrogate of lung hypoplasia, may be useful for risk stratification in CDH. Paco 2 < 60 mm Hg in the first ABG may indicate milder pulmonary hypoplasia. A Paco 2 > 80 mm Hg in the first ABG and/or before ECMO may indicate severe pulmonary hypoplasia.

How Does Ethylenediaminetetraacetic Acid Irrigation Affect Biodentine? A Multimethod Ex Vivo Study.

The activity of biomaterials used during endodontic treatment can be affected by various factors. One of them is the chemical action of the irrigant that they are exposed to. The aim of this multimethod ex vivo study was to evaluate the influence of ethylenediaminetetraacetic acid (EDTA) on the surface appearance and chemical composition of Biodentine used in perforation repair. Twenty material specimens were prepared according to manufacturers' recommendations and divided into two setting-time-based groups, tested after 45 min (group A) and 24 h (group B) of setting. Material was irrigated with 17% EDTA solution with or without simultaneous ultrasonic activation. The surface characteristics and the chemical composition analysis of the Biodentine specimens were performed with the aid of a scanning electron microscope (SEM) and an energy dispersive spectroscopy (EDS) method, respectively. The volumetric loss of material was measured by dedicated digital software in an optical microscope. Statistical analysis was performed. The EDS study confirmed that after the rinsing protocol, the percentage content of elements differed between the groups. The EDTA rinse, whether ultrasonically activated or not, visibly affected the surface appearance and chemical composition of Biodentine. The specimens' surface subjected to irrigation was more irregular under SEM than in a control group. The US activation of the liquid amplified its impact on the tested material. The average volume loss in group A after 5 min irrigation was 3.98 µm3 for each µm2 of the chosen area and it increased up to 7.74 µm3/µm2 after the ultrasonic activation. In group B, indicated volume loss values were 6.30 and 11.70 µm3/µm2 for 5 min irrigation without and with US activation, respectively. Using a 20 min irrigation time and ultrasonic activation increased it up to 32.71 µm3/µm2. Each rinsing protocol involving irrigation with ethylenediaminetetraacetic acid modified the surface features and the chemical composition of the evaluated hydraulic tricalcium silicate cement. Further research is needed to indicate the possible impact of the observed changes on its long-term clinical performance.

The Utility of Intravenous Methylprednisolone as an Adjunct Treatment for Drug-Resistant Amiodarone-Induced Thyrotoxicosis.

BACKGROUND: Amiodarone-induced thyrotoxicosis (AIT) may pose treatment challenges. We present a series of patients in which we achieved the normalisation of free T3 (FT3) using intravenous methylprednisolone (ivMP) in AIT refractory to thiamazole and oral prednisone. Namely, in three males (aged 56, 50 and 64, all with a history of AF and/or a low ejection fraction), an addition of ivMP resulted in the normalisation of FT3, which allowed successful thyroidectomy. In another case of a 65-year-old man, we initially succeeded in the normalisation of FT3 using ivMP from FT4 > 7.77 ng/dL (0.93-1.7) to 2.41 ng/dL and in that of FT3 from 14.95 pg/mL (2-4.4) to 2.05 pg/mL), but four weeks after stopping ivMP, despite the continuation of thiamazole and prednisone, there was rebound thyrotoxicosis: FT4 > 7.77 ng/dL and FT3-5.46 pg/mL. Intravenous MP was restated leading to a decline in FT4 to 2.51 ng/dL and in FT3 to 1.92 pg/mL, thus allowing a successful thyroidectomy. Finally, in a 78-year-old man with AF, goitre, and AIT resistant to thiamazole, prednisone and lithium carbonate, we obtained a reduction in FT4 to 1.51 ng/dL and in FT3 to 3.17 pg/mL after seven pulses of ivMP. Oral prednisone was gradually reduced and successfully stopped about six months later. He remained on low-dose thiamazole (5 mg od). CONCLUSIONS: Pulse ivMP in addition to oral steroids may be a useful adjunct therapy either for the preparation of a thyroidectomy or as a treatment modality in drug-resistant AIT. Though a total cure is possible, there is a danger of a rebound worsening of thyrotoxicosis after premature discontinuation of ivMP.

The legal highs problem in the Polish printed media--actors, claims, and its hidden meanings.

The authors present the results of a qualitative analysis of press articles focused on legal highs in Poland. The aim of analysis was the identification of all social actors involved in the media discourse, the arguments used, and the claims made in the frame of social, political, economical, legal, and moral issues. This analysis covered two major daily newspapers--Gazeta Wyborcza and Rzeczpospolita and two weeklies--Polityka and Newsweek. Articles were collected during a systematic analysis covering the complete number of issues starting from 2008 to 2011. As a result, a base of 386 articles was developed. The study was founded by the statutory budget.

Analysis of Reconstituted Tripartite Complex Supports Avidity-based Recruitment of Hsp70 by Substrate Bound J-domain Protein.

Hsp70 are ubiquitous, versatile molecular chaperones that cyclically interact with substrate protein(s). The initial step requires synergistic interaction of a substrate and a J-domain protein (JDP) cochaperone, via its J-domain, with Hsp70 to stimulate hydrolysis of its bound ATP. This hydrolysis drives conformational changes in Hsp70 that stabilize substrate binding. However, because of the transient nature of substrate and JDP interactions, this key step is not well understood. Here we leverage a well characterized Hsp70 system specialized for iron-sulfur cluster biogenesis, which like many systems, has a JDP that binds substrate on its own. Utilizing an ATPase-deficient Hsp70 variant, we isolated a Hsp70-JDP-substrate tripartite complex. Complex formation and stability depended on residues previously identified as essential for bipartite interactions: JDP-substrate, Hsp70-substrate and J-domain-Hsp70. Computational docking based on the established J-domain-Hsp70(ATP) interaction placed the substrate close to its predicted position in the peptide-binding cleft, with the JDP having the same architecture as when in a bipartite complex with substrate. Together, our results indicate that the structurally rigid JDP-substrate complex recruits Hsp70(ATP) via precise positioning of J-domain and substrate at their respective interaction sites - resulting in functionally high affinity (i.e., avidity). The exceptionally high avidity observed for this specialized system may be unusual because of the rigid architecture of its JDP and the additional JDP-Hsp70 interaction site uncovered in this study. However, functionally important avidity driven by JDP-substrate interactions is likely sufficient to explain synergistic ATPase stimulation and efficient substrate trapping in many Hsp70 systems.

The Effect of Irrigation with Citric Acid on Biodentine Tricalcium Silicate-Based Cement: SEM-EDS In Vitro Study.

There are various factors that may interfere with the activity of biomaterials during endodontic therapy. One of them is the canal system irrigation procedure with different rinsing solutions performed after the placement of bioactive cements. The authors investigate the influence of citric acid, a chelating agent, on the surface and the chemical composition of Biodentine tricalcium silicate-based cement using a multimethod approach. Twenty samples were divided into two groups based on the material setting time. They were subjected to citric acid irrigation with or without ultrasonic activation for 5 and 20 min. The chemical analysis was made with energy dispersive spectroscopy (EDS). The visual assessment of Biodentine surface was carried out in scanning electron microscope (SEM). The volume of material loss during the procedure was measured with Keyence optic microscope and dedicated digital software. Statistical analysis was performed. The results of the study show that the irrigation with citric acid influenced the surface appearance of the material and changed its chemical composition in both investigated groups. The ultrasonic activation (US) of the liquid has also aggravated its impact. Further research is needed to assess if that fact may change the sealing properties of the material influencing the long-term clinical outcome.

Motives for using new psychoactive substances in three groups of Polish users: nightlife, marginalised and active on the In. / Motywy uzywania nowych substancji psychoaktywnych w trzech grupach uzytkowników: rekreacyjnych, zmarginalizowanych i aktywnych w internecie.

OBJECTIVES: The aim of this article is to present the motives for using new psychoactive substances (NPS) among users in Poland and to evaluate the relationships between motives and consumption of different types of NPS. METHODS: The fieldwork was conducted in four locations: in Warsaw, Krakow, Poznan, and Tricity. The study involved a total of 596 users of new psychoactive substances. Among them were: nightlife users, using NPS recreationally (N = 172), socially marginalised users (N = 86) and users active on the internet (N = 338). The technique used in the study was a self-filled questionnaire. RESULTS: As assessed by all respondents, enhancement of mood was the most frequently indicated motive for using NPS. Among nightlife users, the NPS were most often used for the purpose of having more fun at parties. Among the marginalised users, the most common motive for using the new psychoactive substances was the desire to get intoxicated, which is a motive belonging to the group of motives related to enhancement. In the group of people active on the Internet, the most common motives for using these substances were those related to expansion. CONCLUSIONS: Identifying motives for using new psychoactive substances may contribute to reducing the use of NPS. The recreational and marginalised users have different motives for using substances. The same conclusion applies to the use of the individual NPS. The motives of using them vary. Thus, preventive, educational and therapeutic programs should be judiciously adapted to the needs of the users as well as to the kind of substances they use.