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INKA is a test facility designed by Framatome and built in the technical center in Karlstein. The original objective for establishing this test rig was the investigation of the performance of the passive safety systems developed in a new Framatome Boiling Water Reactor (BWR) design – KERENA. INKA was constructed in the scale of 1:1 in heights while the total volume of the containment was replicated in 1:24. Since the geometries of particular safety systems are faithfully reflected, their actual performance in the original plant can be investigated at the full scale. Due to the unquestionable interest of the nuclear community in the inherent safety, not only new BWR and PWR designs are equipped with the passive systems, but also particular passive solutions are considered to be applied into the already existing Light Water Reactors (LWR). In this context and due to the fact that both, single component tests and integral tests can be conducted at INKA, the facility can be employed for a demonstration/qualification of a large range of passive safety systems foreseen for quite different types of LWRs. Hence, the goal of the EASY project was the experimental confirmation of the passive systems performance and the analysis of their interactions between each other in the integral tests. Besides, the overarching target of all tests performed at INKA is provision of data for codes validation. This paper presents major outcomes and conclusions drawn on the basis of EASY project results.

PKL is the only test facility in Europe that replicates the entire primary side and the most important parts of the secondary side of western-type Pressurized Water Reactors (PWR) in the scale of 1:1 in heights. It is also worldwide the only test facility with 4 identical reactor coolant loops arranged symmetrically around the Reactor Pressure Vessel (RPV) for simulation of nonsymmetrical boundary conditions between the reactor loops. Thermal-hydraulic phenomena observed in PWRs are simulated in the PKL test facility for over 40 years. The analyses carried out in these years encompass a large spectrum of accident scenario simulations and corresponding cool-down procedures. The overall goal of the PKL experiments is to show that under accident conditions - even for extreme and highly unlikely assumptions as additional loss of safety systems - the core cooling can be maintained or re-established by automatic or operator- performed procedures and that a severe accident e.g. a core melt-down can be avoided under all circumstances. Another goal of the tests performed in the PKL facility is the provision of data for validation of thermal-hydraulic system codes. This paper presents recent modifications of the PKL facility, applied in order to adapt the facility to the latest western-type designs currently built in the world. The paper discusses also important results obtained in the last years.

The condensation in horizontal tubes plays an important role in determining the operation mode of passive safety systems of modern nuclear power plants. In this paper, two different approaches for modelling of this phenomenon are compared and verified against experimental data. The first approach is based on the flow regime map developed by Tandon. Depending on the regime, the heat transfer coefficient is calculated according to corresponding semi-empirical correlation. The second approach uses a general, fully empirical correlation proposed by Shah. Both models are developed with utilization of the object-oriented, equation-based Modelica language and the open-source Open-Modelica environment. The results are compared with data obtained during a large scale integral test, simulating a Loss of Coolant Accident scenario performed at the dedicated Integral Test Facility Karlstein (INKA) which was built at the Components Testing Department of AREVA in Karlstein, Germany. The INKA facility was designed to test the performance of the passive safety systems of KERENA, the new AREVA boiling water reactor design. INKA represents the KERENA containment with a volume scaling of 1:24. Components heights and levels over the ground are in the full scale. The comparison of simulations results shows a good agreement.

Tuberculosis remains a leading cause of death from a single infection worldwide. Drug resistance to existing and even new antimycobacterials calls for research into novel targets and unexplored mechanisms of action. Recently we reported on the development of tight-binding inhibitors of Mycobacterium tuberculosis (Mtb) lipoamide dehydrogenase (Lpd), which selectively inhibit the bacterial but not the human enzyme based on a differential modality of inhibitor interaction with these targets. Here we report on the striking improvement in inhibitor residence time on the Mtb enzyme associated with scaffold progression from an indazole to 2-cyanoindole. Cryo-EM of Lpd with the bound 2-cyanoindole inhibitor 19 confirmed displacement of the buried water molecule deep in the binding channel with a cyano group. The ensuing hours-long improvement in on-target residence time is associated with enhanced antibacterial activity in axenic culture and in primary mouse macrophages. Resistance to 2-cyanoindole inhibitors involves mutations within the inhibitor binding site that have little effect on inhibitor affinity but change the modality of inhibitor-target interaction, resulting in fast dissociation from Lpd. These findings underscore that on-target residence time is a major determinant of antibacterial activity and in vivo efficacy., (© 2024 Wiley-VCH GmbH.)

Lpd (lipoamide dehydrogenase) in Mycobacterium tuberculosis (Mtb) is required for virulence and is a genetically validated tuberculosis (TB) target. Numerous screens have been performed over the last decade, yet only two inhibitor series have been identified. Recent advances in large-scale virtual screening methods combined with make-on-demand compound libraries have shown the potential for finding novel hits. In this study, the Enamine REAL library consisting of ∼1.12 billion compounds was efficiently screened using the GPU Shape screen method against Mtb Lpd to find additional chemical matter that would expand on the known sulfonamide inhibitor series. We identified six new inhibitors with IC 50 in the range of 5-100 μM. While these compounds remained chemically close to the already known sulfonamide series inhibitors, some diversity was found in the cores of the hits. The two most potent hits were further validated by one-step potency optimization to submicromolar levels. The co-crystal structure of optimized analogue TDI-13537 provided new insights into the potency determinants of the series., Competing Interests: The authors declare no competing financial interest., (© 2023 The Authors. Published by American Chemical Society.)

Tuberculosis remains a leading cause of death from a single bacterial infection worldwide. Efforts to develop new treatment options call for expansion into an unexplored target space to expand the drug pipeline and bypass resistance to current antibiotics. Lipoamide dehydrogenase is a metabolic and antioxidant enzyme critical for mycobacterial growth and survival in mice. Sulfonamide analogs were previously identified as potent and selective inhibitors of mycobacterial lipoamide dehydrogenase in vitro but lacked activity against whole mycobacteria. Here we present the development of analogs with improved permeability, potency, and selectivity, which inhibit the growth of Mycobacterium tuberculosis in axenic culture on carbohydrates and within mouse primary macrophages. They increase intrabacterial pyruvate levels, supporting their on-target activity within mycobacteria. Distinct modalities of binding between the mycobacterial and human enzymes contribute to improved potency and hence selectivity through induced-fit tight binding interactions within the mycobacterial but not human enzyme, as indicated by kinetic analysis and crystallography.

Efforts at host-directed therapy of tuberculosis have produced little control of the disease in experimental animals to date. This is not surprising, given that few specific host targets have been validated, and reciprocally, many of the compounds tested potentially impact multiple targets with both beneficial and detrimental consequences. This puts a premium on identifying appropriate molecular targets and subjecting them to more selective modulation. We discovered an aminopyrimidine small molecule, 2062, that had no direct antimycobacterial activity, but synergized with rifampin to reduce bacterial burden in Mtb infected macrophages and mice and also dampened lung immunopathology. We used 2062 and its inactive congeners as tool compounds to identify host targets. By biochemical, pharmacologic, transcriptomic and genetic approaches, we found that 2062's beneficial effects on Mtb control and clearance in macrophages and in mice are associated with activation of transcription factor EB via an organellar stress response. 2062-dependent TFEB activation led to improved autophagy, lysosomal acidification and lysosomal degradation, promoting bacterial clearance in macrophages. Deletion of TFEB resulted in the loss of IFNγ-dependent control of Mtb replication in macrophages. 2062 also targeted multiple kinases, such as PIKfyve, VPS34, JAKs and Tyk2, whose inhibition likely limited 2062's efficacy in vivo. These findings support a search for selective activators of TFEB for HDT of TB., Competing Interests: The authors have declared that no competing interests exist.

Background: Cytomegalovirus (CMV) can cause severe disease in children and adults with a variety of inherited or acquired T-cell immunodeficiencies, who are prone to multiple infections. It can also rarely cause disease in otherwise healthy persons. The pathogenesis of idiopathic CMV disease is unknown. Inbred mice that lack the gene encoding nitric oxide synthase 2 ( Nos2 ) are susceptible to the related murine CMV infection., Methods: We studied a previously healthy 51-year-old man from Iran who after acute CMV infection had an onset of progressive CMV disease that led to his death 29 months later. We hypothesized that the patient may have had a novel type of inborn error of immunity. Thus, we performed whole-exome sequencing and tested candidate mutant alleles experimentally., Results: We found a homozygous frameshift mutation in NOS2 encoding a truncated NOS2 protein that did not produce nitric oxide, which determined that the patient had autosomal recessive NOS2 deficiency. Moreover, all NOS2 variants that we found in homozygosity in public databases encoded functional proteins, as did all other variants with an allele frequency greater than 0.001., Conclusions: These findings suggest that inherited NOS2 deficiency was clinically silent in this patient until lethal infection with CMV. Moreover, NOS2 appeared to be redundant for control of other pathogens in this patient. (Funded by the National Center for Advancing Translational Sciences and others.)., (Copyright © 2020 Massachusetts Medical Society.)

Heart failure appears in 2% of the adult population in Europe. One in five people aged 40 years will develop heart failure during their lifetime. Heart failure touch 20,000 people in the Opole province. Heart failure is the second, after acute coronary syndromes, urgent cause of admissions to the Clinic of Cardiology at the University Hospital in Opole. The paper presents the prognosis of hospitalization of patients with heart failure for the years 2015-2050 taking into account the processes of depopulation taking place in our region. The analysis makes it possible to predict that the age group particularly exposed to heart failure in the coming decades will be people who today belong to teenagers and young adults. The article presents current methods of treatment of heart failure. Improvement in the prognosis of patients with heart failure can occur through the implementation of the guidelines for treatment of heart failure recommended by the ESC. This goal is to be achieved by introducing the "Comprehensive care for patients with heart failure (KONS)" program in our country. The shift of the burden of care for patient with heart failure to outpatient unit will result in a significant reduction in the number of hospitalizations.

Genetic deficiency of protein kinase R (PKR) in mice was reported to enhance macrophage activation in vitro in response to interferon-γ (IFNγ) and to reduce the burden of Mycobacterium tuberculosis (Mtb) in vivo (Wu et al. PloS One. 2012 7:e30512). Consistent with this, treatment of wild-type (WT) macrophages in vitro with a novel PKR inhibitor (Bryk et al., Bioorg. Med. Chem. Lett. 2011 21:4108-4114) also enhanced IFN-γ-dependent macrophage activation (Wu et al. PloS One. 2012 7:e30512). Here we show that co-treatment with IFN-γ and a new PKR inhibitor identified herein to be highly but not completely selective likewise induced macrophages to produce more reactive nitrogen intermediates (RNI) and tumor necrosis factor alpha (TNF-α) and less interleukin 10 (IL-10) than seen with IFN-γ alone. Unexpectedly, however, this new PKR inhibitor had a comparable effect on PKR-deficient macrophages. Retrospective investigation revealed that the PKR-deficient mice in (Wu et al. PloS One. 2012 7:e30512) had not been backcrossed. On comparing genetically matched PKR-deficient and WT mice, we saw no impact of PKR deficiency on macrophage activation in vitro or during the course of Mtb infection in vivo. In addition, although 129S1/SvImJ macrophage responses to IFN-γ were greater than those of C57BL/6J macrophages, PKR was not required to mediate the IFN-γ-dependent production of IL-10, RNI or TNF-α in either strain. Together the data cast doubt on PKR as a potential therapeutic target for tuberculosis., (© 2018 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Objective: Introduction: Stroke is one of the leading causes of mortality and disability in the world. A wide knowledge of risk factors for cerebrovascular diseases is one of the main elements of primary prevention, while knowledge of stroke symptoms can significantly speed up treatment and reduce negative health effects. The aim: To assess the knowledge about stroke and risk factors of cerebrovascular diseases among the inhabitants of the randomly selected rural community (in Lodz Province)., Patients and Methods: Material and methods: The study included 143 adult residents of Rzasnia. The research was carried out using a self-constructed questionnaire, prepared on the basis of available literature., Results: Results: The average score obtained from the knowledge survey on stroke was 5.79 +/- 3.47 (maximum number of points - 14). The average number of points in people with a family history of stroke was statistically significantly higher (7.46±3.31) than in those with no stroke in the family (4.20±2.98 ) (p <0.001). There was no statistically significant difference between the average number of points and gender (women - 6.13±3.55 points, men - 5.64±3.55, NS) and age (R = -0.1, NS ). However, the positive correlation between educational level and the score obtained in our questionnaire was found (R =0.4, p <0.05). The average number of correctly listed risk factors was: 0.6 ± 1.6. At least one risk factor was reported by only 34% of respondents. Respondents most often mentioned: excessive alcohol consumption (18%), smoking (18%), hypertension (14%), atherosclerosis (11%). The average number of correctly listed symptoms of stroke was: 1.2±1.3. At least one symptom of stroke was reported by only 58% of respondents. The respondents most often reported: headache (29%), speech disorders (22%), gait disturbances (13%), and sensory disorders (13%). If a stroke was suspected, the majority of respondents would call an ambulance (94%)., Conclusion: Conclusions: Knowledge of the residents of the chosen rural commune about the stroke is not satisfactory and should be supplemented with an educational program that would include as many people as possible.

We compared phenotypes of five strains of Mycobacterium tuberculosis (Mtb) differing in their expression of rv1248c and its product, 2-hydroxy-3-oxoadipate synthase (HOAS), with a focus on carbon source-dependent growth rates and attenuation in mice. Surprisingly, an rv1248c transposon mutant on a CDC1551 background grew differently than an rv1248c deletion mutant on the same background. Moreover, the same rv1248c deletion in two different yet genetically similar strain backgrounds (CDC1551 and H37Rv) gave different phenotypes, though each could be complemented. Whole genome re-sequencing did not provide an obvious explanation for these discrepancies. These observations offer a cautionary lesson about the strength of inference from complementation and sequence analysis, and commend consideration of more complex phenomena than usually contemplated in Mtb, such as epigenetic control., (Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Enzymes of central carbon metabolism (CCM) in Mycobacterium tuberculosis (Mtb) make an important contribution to the pathogen's virulence. Evidence is emerging that some of these enzymes are not simply playing the metabolic roles for which they are annotated, but can protect the pathogen via additional functions. Here, we found that deficiency of 2-hydroxy-3-oxoadipate synthase (HOAS), the E1 component of the α-ketoglutarate (α-KG) dehydrogenase complex (KDHC), did not lead to general metabolic perturbation or growth impairment of Mtb, but only to the specific inability to cope with glutamate anaplerosis and nitroxidative stress. In the former role, HOAS acts to prevent accumulation of aldehydes, including growth-inhibitory succinate semialdehyde (SSA). In the latter role, HOAS can participate in an alternative four-component peroxidase system, HOAS/dihydrolipoyl acetyl transferase (DlaT)/alkylhydroperoxide reductase colorless subunit gene (ahpC)-neighboring subunit (AhpD)/AhpC, using α-KG as a previously undescribed source of electrons for reductase action. Thus, instead of a canonical role in CCM, the E1 component of Mtb's KDHC serves key roles in situational defense that contribute to its requirement for virulence in the host. We also show that pyruvate decarboxylase (AceE), the E1 component of pyruvate dehydrogenase (PDHC), can participate in AceE/DlaT/AhpD/AhpC, using pyruvate as a source of electrons for reductase action. Identification of these systems leads us to suggest that Mtb can recruit components of its CCM for reactive nitrogen defense using central carbon metabolites.