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Neovascularization is implicated in the pathology of retinopathy of prematurity (ROP), diabetic retinopathy (DR), and age-related macular degeneration (AMD), which are the leading causes of blindness worldwide. In our work, we analyzed how heme released during hemorrhage affects hypoxic response and neovascularization. Our retrospective clinical analysis demonstrated, that hemorrhage was associated with more severe retinal neovascularization in ROP patients. Our heme-stimulated human retinal pigment epithelial (ARPE-19) cell studies demonstrated increased expression of positive regulators of angiogenesis, including vascular endothelial growth factor-A (VEGFA), a key player of ROP, DR and AMD, and highlighted the activation of the PI3K/AKT/mTOR/VEGFA pathway involved in angiogenesis in response to heme. Furthermore, heme decreased oxidative phosphorylation in the mitochondria, augmented glycolysis, facilitated HIF-1α nuclear translocation, and increased VEGFA/GLUT1/PDK1 expression suggesting HIF-1α-driven hypoxic response in ARPE-19 cells without effecting the metabolism of reactive oxygen species. Inhibitors of HIF-1α, PI3K and suppression of mTOR pathway by clinically promising drug, rapamycin, mitigated heme-provoked cellular response. Our data proved that oxidatively modified forms of hemoglobin can be sources of heme to induce VEGFA during retinal hemorrhage. We propose that hemorrhage is involved in the pathology of ROP, DR, and AMD.

Background Chronic kidney disease (CKD) poses a significant public health challenge globally while impacting patients’ physical function and quality of life. Addressing the issues of physical inactivity and pain management is essential during treatment to improve health-related quality of life. The present study investigated the effect of an aerobic training program with core stabilization exercises for hemodialysis (HD) patients on a transplant waiting list and renal transplant (RTx) patients.Methods A total of 45 patients with CKD were included in the 12-week study: 25 patients receiving HD (12 HD treatment group, 13 HD control group) and 20 patients with RTx (9 RTx treatment group, 11 RTx control group). Functional capacity was measured using the 6-min walk test, pain was measured using the visual analog scale, and health-related quality of life was measured using the Kidney Disease Quality of Life–Short Form 12 questionnaire. Nonparametric statistical tests were performed at a significance level of 0.05.Results Both the HD and RTx treatment groups showed significantly reduced times for the 6-min walking test (p = 0.002 and p = 0.008, respectively), significantly reduced pain severity (p = 0.002 and p = 0.008, respectively), and significantly improved quality of life scores (p = 0.006 and p = 0.041, respectively) by the end of the study compared with control groups.Conclusion Based on the results, structured exercise programs could be effective therapies in CKD management. Therefore, health providers should promote their integration into routine care practices to enhance patient outcomes and well-being.

AbstractIn light chain amyloidosis (LA), the massive glomerular and vascular amyloid deposition leading to interstitial fibrosis/tubular atrophy (IFTA) is thought to be responsible for renal failure. The amyloid deposition in the interstitium and the tubular basement membrane (TBM) has received less attention in the study of LA. We, therefore, collected clinical and laboratory data on patients diagnosed with LA in our Nephrology Department and studied amyloid deposition in the TBM. Twelve LA patients were diagnosed by renal biopsy during a seven-year period. In 4 of the 12, amyloid deposition could also be detected in the TBM. In our first case of a patient with diabetes mellitus, non-amyloid fibrils resembling ‘diabetic fibrillosis’ were also seen by electron microscopy. Despite the double damage, IFTA was negligible, blood vessels were unaffected, and the glomerular deposition was segmental. In the other three cases, significant (>50%) IFTA and a severely reduced estimated glomerular filtration rate were already detected at the time of diagnosis and amyloid deposition was also observed in the blood vessels. These findings indicate the importance of TBM amyloid deposition in the progression of renal disease. This may represent a late-stage presentation of the disease with a heavy LC burden.

Objectives: Recognition of chronic kidney disease (CKD) is crucial in type 2 diabetes mellitus (T2DM). We conducted a nationwide epidemiological study to evaluate T2DM-associated CKD in Hungary between 2016 and 2020.Methods: Annual incidence and prevalence rates of registered CKD amongst all pharmacologically treated T2DM patients were analyzed in different age-groups by the central database of the Hungarian Health Insurance Fund Management. Statistical methods included Poisson regression, Bonferroni test, Chi-square test.Results: We found 499,029 T2DM patients and 48,902 CKD patients in 2016, and 586,075 T2DM patients and 38,347 CKD patients in 2020. The majority of all prevalent T2DM and CKD patients were older (aged 60–69 years: 34.1% and 25.8%; ≥70 years: 36.1% and 64.4%, respectively). The annual incidence of T2DM and incidence rates of CKD in T2DM decreased in 2017–2020 (p < 0.001). The annual prevalence of T2DM increased (p < 0.01), the prevalence rates of CKD in T2DM were low and decreased from 9.8% to 6.5% in 2016–2020 (p < 0.001).Conclusion: Incidence and prevalence of T2DM-associated CKD decreased significantly in Hungary in 2016–2020. Lower prevalence rates of CKD may suggest under-recognition and/or under-reporting.

Hydrogen sulfide (H2S) was previously revealed to inhibit osteoblastic differentiation of valvular interstitial cells (VICs), a pathological feature in calcific aortic valve disease (CAVD). This study aimed to explore the metabolic control of H2S levels in human aortic valves.Lower levels of bioavailable H2S and higher levels of interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α) were detected in aortic valves of CAVD patients compared to healthy individuals, accompanied by higher expression of cystathionine γ-lyase (CSE) and same expression of cystathionine β-synthase (CBS). Increased biogenesis of H2S by CSE was found in the aortic valves of CAVD patients which is supported by increased production of lanthionine. In accordance, healthy human aortic VICs mimic human pathology under calcifying conditions, as elevated CSE expression is associated with low levels of H2S. The expression of mitochondrial enzymes involved in H2S catabolism including sulfide quinone oxidoreductase (SQR), the key enzyme in mitochondrial H2S oxidation, persulfide dioxygenase (ETHE1), sulfite oxidase (SO) and thiosulfate sulfurtransferase (TST) were up-regulated in calcific aortic valve tissues, and a similar expression pattern was observed in response to high phosphate levels in VICs. AP39, a mitochondria-targeting H2S donor, rescued VICs from an osteoblastic phenotype switch and reduced the expression of IL-1β and TNF-α in VICs. Both pro-inflammatory cytokines aggravated calcification and osteoblastic differentiation of VICs derived from the calcific aortic valves. In contrast, IL-1β and TNF-α provided an early and transient inhibition of VICs calcification and osteoblastic differentiation in healthy cells and that effect was lost as H2S levels decreased. The benefit was mediated via CSE induction and H2S generation.We conclude that decreased levels of bioavailable H2S in human calcific aortic valves result from an increased H2S metabolism that facilitates the development of CAVD. CSE/H2S represent a pathway that reverses the action of calcifying stimuli.

Reactive sulfur species (RSS) entail a diverse family of sulfur derivatives that have emerged as important effector molecules in H2S-mediated biological events. RSS (including H2S) can exert their biological roles via widespread interactions with metalloproteins. Metalloproteins are essential components along the metabolic route of oxygen in the body, from the transport and storage of O2, through cellular respiration, to the maintenance of redox homeostasis by elimination of reactive oxygen species (ROS). Moreover, heme peroxidases contribute to immune defense by killing pathogens using oxygen-derived H2O2 as a precursor for stronger oxidants. Coordination and redox reactions with metal centers are primary means of RSS to alter fundamental cellular functions. In addition to RSS-mediated metalloprotein functions, the reduction of high-valent metal centers by RSS results in radical formation and opens the way for subsequent per- and polysulfide formation, which may have implications in cellular protection against oxidative stress and in redox signaling. Furthermore, recent findings pointed out the potential role of RSS as substrates for mitochondrial energy production and their cytoprotective capacity, with the involvement of metalloproteins. The current review summarizes the interactions of RSS with protein metal centers and their biological implications with special emphasis on mechanistic aspects, sulfide-mediated signaling, and pathophysiological consequences. A deeper understanding of the biological actions of reactive sulfur species on a molecular level is primordial in H2S-related drug development and the advancement of redox medicine.

Absztrakt Cél: A tanulmány célja a magyar-ukrán (schengeni külső) határszakaszon 2021-ben végrehajtott Automatizált Határellenőrző Rendszer működési tesztelés-értékelés körülményeinek és eredményeinek bemutatása. A hangsúlyt a rendszer működési hatékonyságára fektettük. Módszertan: Dokumentum- és tartalomelemzés segítségével megvizsgáltuk a rendszer elméleti és jogi hátterét, szabályozási körülményeit, az eredmények elérése érdekében minősítési mérést hajtottunk végre, melynek alapját eszközés módszerspecifikus szempontrendszerek alkották. Megállapítások: A mérések és értékelések során megállapítottuk, hogy a határforgalom- ellenőrzésben alkalmazott humán erőforrás és az ellenőrzés végrehajtási idejének arányát illetően az automatikus határellenőrző rendszernél végrehajtott ellenőrzés hatékonyabbnak bizonyult. Érték: Az Automatizált Határellenőrző Rendszer komponenseit alkotó kioszkok és automatizált átléptető (fix) kapuk hazai tesztelésére és értékelésére schengeni szárazföldi határátkelőhelyen korábban még nem került sor, ezért e vizsgálat és az azon alapuló tanulmány egyedülálló a határrendészeti szcénában.

Introduction: Overweight and obesity are major problems in our society, because it’s prevalence among children and adolescents aged 5-19 has risen dramatically from just 4% in 1975 to just over 18% in 2016. The rise has occurred similarly among both boys and girls: in 2016 18% of girls and 19% of boys were overweight. While just under 1% of children and adolescents aged 5-19 were obese in 1975, more 124 million children and adolescents (6% of girls and 8% of boys) were obese in 2016 (WHO, 2021). Objectives: The aim of our research is to examine the relationship between the time allocated to physical education at school and overweight/obesity in 6-10 year old children on a global and continental level. Methods: Our study is a cross-sectional, population-based descriptive study. We used the data from the World-wide Survey of School Physical Education report published by UNESCO in 2013. We used BMI for the age group 6-10 years from the database published by the Global Burden of Disease Collaborative Network. The GDP data for the countries was taken from the database of the United Nations Economic Commissions for Europe. Results: In our study, we analyze data from 145 countries on six continents. The average time devoted to physical education in these countries is 95.6±35.0 minutes per week. The minimum is 30 minutes and the maximum is 225 minutes. The 90 minutes occurs with the highest frequency, 26 times. The median also shows the 90-minute value. Examining the results by continent, it is revealed that significantly more time is devoted to school physical education in Europe than in Africa (p=0.020) and Asia (p=0.022), but there is no significant difference between the average of the other three continents and Europe. A Spearman’s rank-order correlation was run to assess the relationship between GDP and physical education time allocation. One hundred forty states were included in the analysis. There was a weak positive statistically significant correlation between GDP and physical education time allocation, rs(128) = .298, p < .001. Conclusions: The amount of time devoted to physical education at school shows a wide variation from state to state. The time allocated to physical education at school is 95.6 minutes on average. Europe and Australia/Oceania are the two continents where the most time (about 110 minutes) is devoted to physical education per week. We did not find a relationship between the time devoted to physical education at school and the frequency of obesity or overweight.

Vulnerable atherosclerotic plaques with hemorrhage considerably contribute to cardiovascular morbidity and mortality. Calcification is the main characteristic of advanced atherosclerotic lesions and calcified aortic valve disease (CAVD). Lyses of red blood cells and hemoglobin (Hb) release occur in human hemorrhagic complicated lesions. During the interaction of cell-free Hb with plaque constituents, Hb is oxidized to ferric and ferryl states accompanied by oxidative changes of the globin moieties and heme release. Accumulation of both ferryl-Hb and metHb has been observed in atherosclerotic plaques. The oxidation hotspots in the globin chain are the cysteine and tyrosine amino acids associated with the generation of Hb dimers, tetramers and polymers. Moreover, fragmentation of Hb occurs leading to the formation of globin-derived peptides. A series of these pro-atherogenic cellular responses can be suppressed by hydrogen sulfide (H2S). Since H2S has been explored to exhibit a wide range of physiologic functions to maintain vascular homeostasis, it is not surprising that H2S may play beneficial effects in the progression of atherosclerosis. In the present review, we summarize the findings about the effects of H2S on atherosclerosis and CAVD with a special emphasis on the oxidation of Hb/heme in atherosclerotic plaque development and vascular calcification.

Cél: A határrendészeti tisztképzés elmúlt harminc évi eredményének összefoglalása mellett elődlegesen a jelenlegi társadalmi és szakmai kihívásoknak megfelelő képzési módszerek, nehézségek, kihívások bemutatása, illetve a jövőbeni képzési rendszerre vonatkozó elképzelések vázolása. A jövőbeni határrendészeti tisztképzés alapjait képező Magyarország Nemzeti Integrált Határigazgatási Stratégiájában foglaltak és a határrendészeti tevékenység globális, európai uniós és kontinentális szerepének összevetése egy új, gyakorlatorientált képzési forma bemutatásával. Kiemelt cél olyan újszerű technológiák oktatásban történő bemutatása, mint a biometria, mesterséges intelligencia vagy a robotika. Módszertan: A publikáció elsődlegesen a tapasztalatok alapján összegyűjtött eredmények feldolgozására kíván fókuszálni, illetve e mellett az Európai Unió és a hazai határrendészeti kihívások számbavételével próbálja megközelíteni a határrendészeti tisztképzés jelenét és jövőjét. Megállapítások: A határrendészeti tisztképzés folyamatosan fejlődött az elmúlt harminc évben és ezt a fejlődést érdemes tovább gondozni. A cél azonban már az, hogy új módszerekkel gazdagítsuk az oktatói repertoárt, és egyértelműen nagyobb hangsúlyt kell fektetni a nemzetközi tapasztalatok feldolgozására. Érték: Az egyetemi határrendészeti tiszti oktatás eredményeként olyan menedzserszemléletű vezetők képezhetők, akik a jövőbeni kihívások esetén is képesek lesznek önállóan megállni helyüket nemcsak az államhatárhoz kapcsolódó, hanem a schengeni térség teljes mélységében zajló, a biztonság és jog térségéhez kötődő feladatok tekintetében is.

Less invasive surfactant administration techniques, together with nasal continuous airway pressure (LISA-nCPAP) ventilation, an emerging noninvasive ventilation (NIV) technique in neonatology, are gaining more significance, even in extremely premature newborns (ELBW), under 27 weeks of gestational age. In this review, studies on LISA-nCPAP are compiled with an emphasis on short- and long-term morbidities associated with prematurity. Several perinatal preventative and therapeutic investigations are also discussed in order to start integrated therapies as numerous organ-saving techniques in addition to lung-protective ventilations. Two thirds of immature newborns can start their lives on NIV, and one third of them never need mechanical ventilation. With adjuvant intervention, these ratios are expected to be increased, resulting in better outcomes. Optimized cardiopulmonary transition, especially physiologic cord clamping, could have an additively beneficial effect on patient outcomes gained from NIV. Organ development and angiogenesis are strictly linked not only in the immature lung and retina, but also possibly in the kidney, and optimized interventions using angiogenic growth factors could lead to better morbidity-free survival. Corticosteroids, caffeine, insulin, thyroid hormones, antioxidants, N-acetylcysteine, and, moreover, the immunomodulatory components of mother’s milk are also discussed as adjuvant treatments, since immature newborns deserve more complex neonatal interventions.

Background: Recent genome-wide association studies demonstrated the association between the prevalence of chronic kidney disease (CKD) and rs11959928, rs626277, and rs7805747 polymorphisms. Materials and Methods: In this study, we investigated the association between CKD and these polymorphisms in patients and controls according to gender. High-resolution melting analysis was performed to detect DAB2 rs11959928, DACH1 rs626277, and PRKAG2 rs7805747 single nucleotide polymorphisms. Genomic DNA was extracted from the buffy coat of 163 patients with chronic renal disease and 218 control individuals. Ten percent of the results were also randomly confirmed by direct DNA sequencing. Results: Multivariable logistic regression analysis with adjustment for confounders showed rs7805747 (dominant model) has a statistically significant protective effect in females, and rs11959928 (additive and dominant models) was significantly associated with the prevalence of CKD in males. rs7805747 (recessive model) was significantly associated with the prevalence of CKD in males. Conclusion: The very same genetic variants have different effects in males and females separately. Our results warrant the need for similar studies in larger cohorts.

Abstract Hemorrhage and hemolysis with subsequent heme release are implicated in many pathologies. Endothelial cells (ECs) encounter large amount of free heme after hemolysis and are at risk of damage from exogenous heme. Here we show that hemorrhage aggravates endoplasmic reticulum (ER) stress in human carotid artery plaques compared to healthy controls or atheromas without hemorrhage as demonstrated by RNA sequencing and immunohistochemistry. In EC cultures, heme also induces ER stress. In contrast, if cultured ECs are pulsed with heme arginate, cells become resistant to heme-induced ER (HIER) stress that is associated with heme oxygenase-1 (HO-1) and ferritin induction. Knocking down HO-1, HO-2, biliverdin reductase, and ferritin show that HO-1 is the ultimate cytoprotectant in acute HIER stress. Carbon monoxide-releasing molecules (CORMs) but not bilirubin protects cultured ECs from HIER stress via HO-1 induction, at least in part. Knocking down HO-1 aggravates heme-induced cell death that cannot be counterbalanced with any known cell death inhibitors. We conclude that endothelium and perhaps other cell types can be protected from HIER stress by induction of HO-1, and heme-induced cell death occurs via HIER stress that is potentially involved in the pathogenesis of diverse pathologies with hemolysis and hemorrhage including atherosclerosis.

Macrophages express the A subunit of coagulation factor XIII (FXIII-A), a transglutaminase which cross-links proteins through Nε-(γ-L-glutamyl)-L-lysyl iso-peptide bonds. Macrophages are major cellular constituents of the atherosclerotic plaque; they may stabilize the plaque by cross-linking structural proteins and they may become transformed into foam cells by accumulating oxidized LDL (oxLDL). The combination of oxLDL staining by Oil Red O and immunofluorescent staining for FXIII-A demonstrated that FXIII-A is retained during the transformation of cultured human macrophages into foam cells. ELISA and Western blotting techniques revealed that the transformation of macrophages into foam cells elevated the intracellular FXIII-A content. This phenomenon seems specific for macrophage-derived foam cells; the transformation of vascular smooth muscle cells into foam cells fails to induce a similar effect. FXIII-A containing macrophages are abundant in the atherosclerotic plaque and FXIII-A is also present in the extracellular compartment. The protein cross-linking activity of FXIII-A in the plaque was demonstrated using an antibody labeling the iso-peptide bonds. Cells showing combined staining for FXIII-A and oxLDL in tissue sections demonstrated that FXIII-A-containing macrophages within the atherosclerotic plaque are also transformed into foam cells. Such cells may contribute to the formation of lipid core and the plaque structurization.

Introduction: Hydrogen sulfide (H2S) was revealed to inhibit aortic valve calcification and inflammation was implicated in the pathogenesis of calcific aortic valve disease (CAVD). Objectives: We investigate whether H2S inhibits mineralization via abolishing inflammation. Methods and results: Expression of pro-inflammatory cytokines, interleukin-1β (IL-1β) and tumor necrosis factor α (TNF-α) were increased in patients with CAVD and in calcified aortic valve of ApoE-/- mice. Administration of H22S releasing donor (4-methoxyphenyl piperidinylphosphinodithioc acid (AP72)) exhibited inhibition on both calcification and inflammation in aortic valve of apolipoprotein E knockout mice (ApoE-/-) mice is reflected by lowering IL-1β and TNF-α levels. Accordingly, AP72 prevented the accumulation of extracellular calcium deposition and decreased nuclear translocation of nuclear factor-κB (NF-κB) in human valvular interstitial cells (VIC). This was also accompanied by reduced cytokine response. Double-silencing of endogenous H2S producing enzymes, Cystathionine gamma-lyase (CSE) and Cystathionine beta-synthase (CBS) in VIC exerted enhanced mineralization and higher levels of IL-1β and TNF-α. Importantly, silencing NF-κB gene or its pharmacological inhibition prevented nuclear translocation of runt-related transcription factor 2 (Runx2) and subsequently the calcification of human VIC. Increased levels of NF-κB and Runx2 and their nuclear accumulation occurred in ApoE-/- mice with a high-fat diet. Administration of AP72 decreased the expression of NF-κB and prevented its nuclear translocation in VIC of ApoE-/- mice on a high-fat diet, and that was accompanied by a lowered pro-inflammatory cytokine level. Similarly, activation of Runx2 did not occur in VIC of ApoE-/- mice treated with H2S donor. Employing Stimulated Emission Depletion (STED) nanoscopy, a strong colocalization of NF-κB and Runx2 was detected during the progression of valvular calcification. Conclusions: Hydrogen sulfide inhibits inflammation and calcification of aortic valve. Our study suggests that the regulation of Runx2 by hydrogen sulfide (CSE/CBS) occurs via NF-κB establishing a link between inflammation and mineralization in vascular calcification.

The introduction of Bruton's tyrosine kinase inhibitor ibrutinib has made a significant progress in the treatment of chronic lymphocytic leukemia and other B-cell malignancies. Due to the reduction of cytokine release, it is effective in chronic graft-versus-host disease, and its use has also been suggested in autoimmune diseases and in prevention of COVID-19-associated lung damage. Despite this effect on the immune response, we report a severe hypersensitivity reaction in a 76-year-old male patient diagnosed with prolymphocytic leukemia. Four weeks after the ibrutinib start, non-oliguric acute kidney injury with proteinuria and microscopic hematuria developed and that was accompanied by lower limb purpuras and paresthesia. Renal biopsy revealed acute interstitial nephritis. Employing 1 mg/kg methylprednisolone administration, serum creatinine decreased from 365 μmol/L to 125 μmol/L at 11 days and the proteinuria-hematuria as well as the purpura, paresthesia resolved. Three months later at stabile eGFR of 56 ml/min/1.73 m2 methylprednisolone was withdrawn and a rituximab-venetoclax treatment was initiated without side effects. We conclude that despite the beneficial effect on cytokines response in Th1 direction, ibrutinib can cause acute interstitial nephritis. Early detection, discontinuation of ibrutinib, glucocorticoid administration may help to better preserve renal function, thereby lowering the risk of potential subsequent kidney injury.

Chronic kidney disease (CKD) is a significant public health challenge with a substantial associated risk of mortality, morbidity, and health care expenditure. Culprits that lead to development and progression of CKD are multifaceted and heterogenous in nature. This notion underscores the need for diversification of animal models to investigate its pathophysiology, related complications, and to subsequently enable discovery of novel therapeutics. Importantly, animal models that could recapitulate complications of CKD in both genders are desperately needed. Cardiovascular disease is the most common cause of death in CKD patients that may be due in part to high prevalence of vascular calcification (VC). Using DBA/2 mice that are susceptible to development of VC, we sought to investigate the feasibility and reproducibility of a unilateral ischemia-reperfusion model followed by contralateral nephrectomy (UIRI/Nx) to induce CKD and its related complications in female and male mice. Our results demonstrate that irrespective of gender, mice faithfully displayed complications of moderate CKD following UIRI/Nx as evidenced by significant rise in serum creatinine, albuminuria, higher degree of collagen deposition, elevated expression of classic fibrotic markers, higher circulating levels of FGF-23, PTH and hepcidin. Moreover, we corroborate the osteoblastic transition of aortic smooth muscle cells and cardiomyocytes based on higher levels of osteoblastic markers namely, Cbfa-1, osteopontin, osteocalcin, and osterix. Our data confirms a viable, and consistent model of moderate CKD and its associated complications in both male and female mice. Furthermore, early evidence of osteoblastic transition of cardiovascular system in this model confirms its suitability for studying and implementing potential preventive and/or therapeutic approaches that are urgently needed in this field.

Abstract Background Wolcott-Rallison Syndrome (WRS) is a rare autosomal recessive disease that is the most common cause of neonatal diabetes in consanguineous families. WRS is caused by various genetic alterations of the Eukaryotic Translation Initiation Factor 2-Alpha Kinase 3 (EIF2AK3) gene. Methods Genetic analysis of a consanguineous family where two children were diagnosed with WRS was performed by Sanger sequencing. The altered protein was investigated by in vitro cloning, expression and immunohistochemistry. Results The first cases in Hungary, − two patients in one family, where the parents were fourth-degree cousins - showed the typical clinical features of WRS: early onset diabetes mellitus with hyperglycemia, growth retardation, infection-induced multiple organ failure. The genetic background of the disease was a novel alteration in the EIF2AK3 gene involving the splice site of exon 11– intron 11–12 boundary: g.53051_53062delinsTG. According to cDNA sequencing this created a new splice site and resulted in a frameshift and the development of an early termination codon at amino acid position 633 (p.Pro627AspfsTer7). Based on in vitro cloning and expression studies, the truncated protein was functionally inactive. Immunohistochemistry revealed that the intact protein was absent in the islets of pancreas, furthermore insulin expressing cells were also dramatically diminished. Elevated GRP78 and reduced CHOP protein expression were observed in the liver. Conclusions The novel genetic alteration causing the absence of the EIF2AK3 protein resulted in insufficient handling of severe endoplasmic reticulum stress, leading to liver failure and demise of the patients.

Given the abundance of heme proteins (cytochromes) in the mitochondrion, it is evident that a meticulously orchestrated iron metabolism is essential for cardiac health. Here, we examined the functional significance of myocardial ferritin heavy chain (FtH) in a model of acute myocardial infarction. We report that FtH deletion did not alter either the mitochondrial regulatory and surveillance pathways (fission and fusion) or mitochondrial bioenergetics in response to injury. Furthermore, deletion of myocardial FtH did not affect cardiac function, assessed by measurement of left ventricular ejection fraction, on days 1, 7, and 21 post injury. To identify the modulated pathways providing cardiomyocyte protection coincident with FtH deletion, we performed unbiased transcriptomic analysis. We found that following injury, FtH deletion was associated with upregulation of several genes with anti-ferroptotic properties, including heme oxygenase-1 (HO-1) and the cystine/glutamate anti-porter (Slc7a11). These results suggested that HO-1 overexpression mitigates ferroptosis via upregulation of Slc7a11. Indeed, using transgenic mice with HO-1 overexpression, we demonstrate that overexpressed HO-1 is coupled with increased Slc7a11 expression. In conclusion, we demonstrate that following injury, myocardial FtH deletion leads to a compensatory upregulation in a number of anti-ferroptotic genes, including HO-1. Such HO-1 induction leads to overexpression of Slc7a11 and protects the heart against ischemia-reperfusion-mediated ferroptosis, preserves mitochondrial function, and overall function of the myocardium.

Introduction. The early ages of adolescence, between 11-14, are accompanied by a great fluctuation of the values of sudden physical growth, of temperament and of personality. Periodical monitoring of somatic indicators and determining the body mass index are methods necessary for monitoring the state of health of school children. Goals. The goal of this study is to assess the prevalence of overweight and obesity among children in middle school within Bihor, Cluj and Sălaj county and to compare the obtained results with the results of other studies of this nature. Subjects and methods. The sample group was made up of 962 middle school students. The method used was anthropometry. The body mass index was calculated in accordance with the BMI reference chart according to age and gender. The results of the measurements were statistically processed with the SPSS program. The descriptive analysis was performed and the differences between the average values were tested with the independent samples t-test. We calculated the relation between different variables using the Pearson correlation coefficient. Results. In this study, the prevalence of overweight and obesity were 15.1% respectively 9.8%. There was a significant positive correlation between age and BMI, r = 0.15, p < 0.001, but a negative correlation between age and physical activity, r = - 0.25, p < 0.001. Conclusions. The prevalence of overweight and obesity was 24.9%. The prevalence of overweight and obesity is higher in boys than in girls. The prevalence of overweight is higher in urban boys than in rural boys. REZUMAT. Incidența supraponderalității și obezității la copiii din ciclul gimnazial din România. Introducere. Parcurgerea adolescenței timpurii, cu limite între 11 și 14 ani, se produce cu o mare variabilitate a indicilor creșterii fizice bruște, a temperamentului și a personalității. Supravegherea periodică a indicatorilor somatici și determinarea valorii indicelui de masă corporală reprezintă metode necesare de monitorizare a stării de sănătate a școlarilor. Obiective. Obiectivul acestui studiu a fost să evaluăm incidența supraponderalității și obezității la copii gimnaziali din județele Bihor, Cluj și Sălaj și să comparăm rezultatele obținute cu cele din alte studii asemănătoare. Subiecți și metode. Eșantionul a fost format din 962 elevi din ciclul gimnazial. Metoda de lucru a fost antropometria. Indicele de masă corporală s-a calculat conform hărții de referință a IMC după vârstă și gen. Datele măsurătorilor au fost prelucrate statistic cu ajutorul programului SPSS. A fost făcută analiza descriptivă iar diferențele dintre medii au fost testate cu ajutorul testului t pentru eșantioane independente. Legătura dintre variabile a fost calculat cu ajutorul coeficientului de corelație Pearson. Rezultate. În studiul prezent prevalența supraponderalității a fost de 15,1% iar cea a obezității de 9,8%. A existat o corelație pozitivă semnificativă între vârstă și IMC, r = 0,15, p

Az MTA Bölcsészettudományi Kutatóközpont Történettudományi Intézet színvonalas kiadványsorozatában jelent meg 2019-ben Bencsik Péter határrendészetet is érintő kötete. Az egységes kinézetű, egyszerű, ugyanakkor rendkívül gusztusos (amennyiben könyvről ilyen jelzőt szabad leírni) megjelenésű könyv illik a sorozatba. Ugyanaz a kellemes szürkés borító, az igényes keménytáblás védelem és az elmaradhatatlan három kis képecske teszi vonzóvá és a sorozatot gyűjtők számára keresetté az új kiadványt. A könyv találóan a Kelet és Nyugat között. Államhatárok, úti okmányok, határátlépés Magyarországon és Csehszlovákiában (1945–1989) címet kapta. Ami a rendészeti területen dolgozókat először megfogja a könyv kézbevételekor az a könyv borítóját színesítő (bár fekete-fehér) kis képek, amelyek egy-egy életképet mutatnak be az egykori magyar-csehszlovák határviszonylatban végrehajtott határforgalom-ellenőrzésekhez kapcsolódóan.

Introduction: Climbing consists in raising or moving the body using the arms or the arms and legs from a suspension or a holding position...climbing with the arms and legs recruits the muscles of the entire body, in particular the core and upper limbs. Objectives: Our primary goal is to find out how often the subjects from Romania are engaged in climbing activities and what the most popular types of climbing are. Methods: Throughout the school period of March 4, 2019 to March 11, 2019, 83 (46 boys with a mean age of 12.84±1.24, and 37 girls with a mean age of 12.83±1. 23) children were examined. 68.7% of the children were urban residents and 31.3% were rural residents. Results: On a weekly basis, 14.5% of respondents experience some form of climbing. The 64% of respondents say that their school gym has equipment (rope, rod, ribbed wall) that is suitable for practicing climbing. 89.2% of respondents have been climbing a forest adventure park at least once. However, 30% of these have been at least 5 times and another 35% 3-4 times in an adventure park. Discussion: Climbing is a movement skill that needs to be taught. Its teaching is prescribed in current and older Romanian school curricula. The teaching of its simpler forms in almost every school could be possible by a physical educator or teacher. It would be important to teach and practice as it has many benefits for children, as studies have shown. Conclusion: The interviewed children have already tried several types of climbing, among which tree climbing dominates. Of the climbs which use safety equipment, indoor and obstacle course climbs are more common. REZUMAT. Prevalența activităților de cățărare la copii de 11-15 ani din România: un studiu pilot transversal. Introducere: cățărarea constă în ridicarea și deplasarea corpului folosind doar brațele sau simultan membrele inferioare și superioare dintr-o poziție suspendată. În cățărare sunt angrenați toți mușchii corpului, mai ales ai brațelor și ai trunchiului. Obiective: obiectivul principal al acestui studiu este de a afla cât de frecvent sunt implicați subiecții din România în activități de cățărare și care sunt cele mai populare tipuri de cățărare. Metode: în perioada 4 martie 2019 - 11 martie 2019, au fost examinați 83 de copii (46 de băieți cu o vârstă medie de 12,84 ± 1,24 ani și 37 de fete cu o vârstă medie de 12,83 ± 1,23 ani). 68,7% dintre copii erau din mediul urban și 31,3% din mediul rural. Rezultate: 14.5% dintre copii practică cu frecvență săptămânală cel puțin o formă a cățărării. 64% dintre ei au răspuns că sala lor de sport este dotată cu diferite echipamente pentru practicarea cățărării (frânghie, bară, spalier). 89.2% dintre copii au fost deja într-un parc de aventură forestier. Discuții: cățărarea este o deprindere motrică care trebuie învățată. Predarea acesteia a fost și este prevăzută în programele școlare actuale și cele vechi. Învățarea formelor simple ale cățărârii ar putea fi posibilă de către un profesor de educație fizică sau de învățătoarele claselor primare. Practicarea acesteia ar fi importantă, deoarece are numeroase beneficii fizice pentru practicanți, așa cum au arătat unele studii. Concluzii: copiii intervievați au încercat deja mai multe tipuri de cățărare, printre care domină cățărarea pe copaci. Dintre acele tipuri care folosesc echipamente de siguranță sunt mai frecvente cățărările la sală și cele pe obstacole în parcuri de aventură. Cuvinte cheie: cățărare, cățărare pe obstacole, parc de aventură forestier, activitate fizică.

The long-term effects of low-density lipoprotein (LDL) apheresis (LA) on the progression and regression of atherosclerosis were evaluated by carotid ultrasonography in two severe heterozygous familial hypercholesterolemic patients with carotid artery stenosis. Furthermore, the lipid-lowering efficacy of LA was evaluated. A 63-year-old male and a 64-year-old female patient have been followed for severe heterozygous FH, being treated by LA for 5 years continuously, at 4-week intervals, added on a standard high-dose statin and ezetimibe therapy. The regular LA treatment significantly decreased the levels of total cholesterol (by −58.1 and −53.8%) and LDL-C (by −69 and −66%), while the HDL-C levels did not change significantly. The time-averaged LDL-C (TAL) levels were also significantly decreased (by 20.8 and 30.1%). Still, we could not achieve the 1.8 mmol/L LDL-C goal values (average TAL levels were 4.1 and 5.1 mmol/L). Carotid ultrasonography was performed in every 6 months; internal carotid arteries (ICA) and common carotid arteries (CCA) were evaluated on both sides. Although during the 5-year follow-up we found a progression in the left ICA of the male patient (from 20% to 45%), all the other parameters showed the slow up of the atherogenesis in the studied arteries. The results suggest that monthly LA-treatment can significantly delay the progression of the atherosclerotic process in FH patients, however, it is not adequate to stop progression in every case because of TAL levels exceeding the LDL-C goal value. LA treatment twice a month or PCSK9 inhibitor therapy should be considered to further lower LDL-C.

Vascular calcification is a life-threatening clinical condition in chronic kidney disease (CKD) and is associated with reduced zinc serum levels. Anemia is another frequent complication of CKD. Hypoxia-inducible factor (HIF) stabilizers, also known as HIF prolyl hydroxylase inhibitors (PHI), are promising candidates to treat CKD-associated anemia by increasing erythropoietin synthesis. Recent evidence suggests that HIFs play a pivotal role in vascular calcification. Our study explored feasible impacts of HIF PHI on phosphate (Pi)-induced calcification of vascular smooth muscle cells (VSMCs) and tested whether zinc might inhibit this mineralization process. Treatment of VSMCs with PHI aggravated Pi-induced calcium deposition and Pi uptake. PHI promoted Pi-induced loss of smooth muscle cell markers (ACTA-2, MYH11, SM22α) and enhanced osteochondrogenic gene expression (Msx-2, BMP-2, Sp7) triggering osteochondrogenic phenotypic switch of VSMCs. These effects of PHI paralleled with increased pyruvate dehydrogenase kinase 4 (PDK4) expression, decreased Runx2 Ser451 phosphorylation, and reduced cell viability. Zinc inhibited Pi-induced mineralization of VSMCs in a dose-dependent manner and also attenuated the pro-calcification effect of PHI in Pi-induced mineralization. Zinc inhibited osteochondrogenic phenotypic switch of VSMCs reflected by lowering Pi uptake, decreasing the expressions of Msx-2, BMP-2, and Sp7 as well as the loss of smooth muscle cell-specific markers. Zinc preserved phosphorylation state of Runx2 Ser451, decreased PDK4 level, and restored cell viability. PHI alone reduced the expression of smooth muscle markers without inducing mineralization, which was also inhibited by zinc. In addition, we observed a significantly lower serum zinc level in CKD as well as in patients undergoing carotid endarterectomy compared to healthy individuals. Conclusion - PHI promoted the loss of smooth muscle markers and augmented Pi-induced osteochondrogenic phenotypic switch leading to VSMCs calcification. This mineralization process was attenuated by zinc. Enhanced vascular calcification is a potential risk factor during PHI therapy in CKD which necessitates the strict follow up of vascular calcification and zinc supplementation.

Vascular calcification associated with high plasma phosphate (Pi) level is a frequent complication of hyperglycemia, diabetes mellitus, and chronic kidney disease. BGP-15 is an emerging anti-diabetic drug candidate. This study was aimed to explore whether BGP-15 inhibits high Pi-induced calcification of human vascular smooth muscle cells (VSMCs) under normal glucose (NG) and high glucose (HG) conditions. Exposure of VSMCs to Pi resulted in accumulation of extracellular calcium, elevated cellular Pi uptake and intracellular pyruvate dehydrogenase kinase-4 (PDK-4) level, loss of smooth muscle cell markers (ACTA, TAGLN), and enhanced osteochondrogenic gene expression (KLF-5, Msx-2, Sp7, BMP-2). Increased Annexin A2 and decreased matrix Gla protein (MGP) content were found in extracellular vesicles (EVs). The HG condition markedly aggravated Pi-induced VSMC calcification. BGP-15 inhibited Pi uptake and PDK-4 expression that was accompanied by the decreased nuclear translocation of KLF-5, Msx-2, Sp7, retained VSMC markers (ACTA, TAGLN), and decreased BMP-2 in both NG and HG conditions. EVs exhibited increased MGP content and decreased Annexin A2. Importantly, BGP-15 prevented the deposition of calcium in the extracellular matrix. In conclusion, BGP-15 inhibits Pi-induced osteochondrogenic phenotypic switch and mineralization of VSMCs in vitro that make BGP-15 an ideal candidate to attenuate both diabetic and non-diabetic vascular calcification.

With iron at its core, the tetrapyrrole heme ring is a cardinal prosthetic group made up of many proteins that participate in a wide array of cellular functions and metabolism. Once released, due to its pro-oxidant properties, free heme in sufficient amounts can result in injurious effects to the kidney and other organs. Heme oxygenase-1 (HO-1) has evolved to promptly attend to such injurious potential by facilitating degradation of heme into equimolar amounts of carbon monoxide, iron, and biliverdin. HO-1 induction is a beneficial response to tissue injury in diverse animal models of diseases, including those that affect the kidney. These protective attributes are mainly due to: (i) prompt degradation of heme leading to restraining potential hazardous effects of free heme, and (ii) generation of byproducts that along with induction of ferritin have proven beneficial in a number of pathological conditions. This review will focus on describing clinical aspects of some of the conditions with the unifying end-result of increased heme burden and will discuss the molecular mechanisms that ensue to protect the kidneys.

Infiltration of red blood cells into atheromatous plaques and oxidation of hemoglobin (Hb) and lipoproteins are implicated in the pathogenesis of atherosclerosis. α1-microglobulin (A1M) is a radical-scavenging and heme-binding protein. In this work, we examined the origin and role of A1M in human atherosclerotic lesions. Using immunohistochemistry, we observed a significant A1M immunoreactivity in atheromas and hemorrhaged plaques of carotid arteries in smooth muscle cells (SMCs) and macrophages. The most prominent expression was detected in macrophages of organized hemorrhage. To reveal a possible inducer of A1M expression in ruptured lesions, we exposed aortic endothelial cells (ECs), SMCs and macrophages to heme, Oxy- and FerrylHb. Both heme and FerrylHb, but not OxyHb, upregulated A1M mRNA expression in all cell types. Importantly, only FerrylHb induced A1M protein secretion in aortic ECs, SMCs and macrophages. To assess the possible function of A1M in ruptured lesions, we analyzed Hb oxidation and heme-catalyzed lipid peroxidation in the presence of A1M. We showed that recombinant A1M markedly inhibited Hb oxidation and heme-driven oxidative modification of low-density lipoproteins as well plaque lipids derived from atheromas. These results demonstrate the presence of A1M in atherosclerotic plaques and suggest its induction by heme and FerrylHb in the resident cells.

A schengeni eszme egyik meghatározó vívmánya, hogy a személyek, a járművek és szállítmányok szabadon, ellenőrzés nélkül mozoghatnak a részes tagállamok belső határain. Amennyiben a schengeni külső határok ellenőrzési mechanizmusa nem garantálja a térség biztonságát, illetve egy tagállam a saját biztonsága érdekében dönt a schengeni belső határain a határellenőrzés ideiglenes visszaállításáról, akkor annak mind személyzeti, mind technikai vonzatával számolni kell. A tanulmány fő célja az ellenőrzés idejének, helyének, tartalmának, humánerőforrás igényének, valamint technikai és informatikai hátterének vizsgálata.

Background: Depending on the severity, the nature and the type of the trunk asymmetry, three types of treatment are currently used: (1) conservative treatment with orthopedic brace, (2) physiotherapy and (3) surgical intervention. The efficiency of physical exercises in rehabilitation of spinal deformities is well known. They have been systematically used since the beginning of the 20th century. Objectives: The main objective of our study was to reduce the trunk asymmetry using an interventional program of physical exercises. Study design: This study follows a pretest-posttest design with an experimental and a control group. Materials and Methods: Forty one pupils (27 girls and 14 boys) aged 11.77 (± 0.96) years) were included. The experimental group consisted of 20 pupils and the control of 21 students. Participants were assigned non-randomly to the experiment or control groups. In order to determine the level of physical activity of the subjects, the FELS and PAQ-C questionnaires were applied. Experimental intervention consisted of a physical activity program based on basic gymnastics. Between October 2015 and June 2016 the members of the experimental group participated in our intervention twice a week (50 minutes a session), after their school program, for 8 months. Results: The mean value of the angle of trunk rotation in the case of experimental group was 6.15º (± 1.56) and 6.52º (± 1.66) in the case of control group. In the final test, it was found that the size of the asymmetry was reduced in both groups. In the case of experimental group the trunk asymmetry value was 4.05º (± 1.66), and in the case of control group 6.00º (± 1.94), meaning a decrease of 2.10º to experimental group and 0.52º to control group. Conclusions: The main findings are that basic gymnastics exercises can have a beneficial effect on trunk asymmetry, which is the hump will decrease. We recommend the use of these exercises to treat people with moderate or severe trunk asymmetry. REZUMAT. Diminuarea asimetriei toracelui la adolescenti folosind un program de intervenție bazate pe exerciții fizice. În funcție de gravitatea, natura și tipul asimetriei toracelui, în prezent sunt utilizate trei tipuri de tratament: tratamentul ortopedic prin utilizarea corsetelor corective, tratamentul cinetic și cel chirurgical. Eficiența exercițiilor fizice în reabilitarea deformărilor coloanei vertebrale este bine cunoscută. Acestea au fost folosite sistematic de la începutul secolului al XX-lea. Obiective: Obiectivul principal al studiului a fost diminuarea asimetriei toracelui folosind un program de intervenție bazate pe exerciții fizice. Study design: Studiul are un design experimental cu un grup experimental și unul de control și cu testare inițială și finală. Materiale și metode: Patruzeci și unu de elevi (27 de fete și 14 băieți) cu vârsta de 11.77±(0.96) ani au fost incluși în studiu. Grupa experimentală a fost alcătuit din 20 de elevi și cel de control din 21 de elevi. Participanții au fost repartizați în mod nealeatoriu în grupa experimentală sau cea de control. Pentru a determina nivelul activității fizice al subiecților au fost aplicate chestionarele FELS ȘI PAQ-C. Intervenția experimentală a constat dintr-un program de activitate fizică bazat pe mijloacele gimnasticii de bază. Între octombrie 2015 și iunie 2016, membrii grupei experimentale au participat la intervenție de două ori pe săptămână (câte 50 de minute pe fiecare sesiune), după programul școlar al elevilor, timp de 8 luni. Rezultate: Valoarea mediei unghiul de rotație al trunchiul la testarea inițială la Ge a fost de 6.15º (±1.56) și de 6.52º (±1.66) în cazul Gc. La finalul intervenției la ambele grupe s-a diminuat mărimea asimetriei, în cazul Ge rezultatul post-testului fiind de 4.05º (±1.66), iar în cazul Gc de 6.00º (±1.94), însemnând o diminuare de 2.10º la Ge și de 0.52º la cel de control. Concluzii: Constatările principale sunt că exercițiile fizice din gimnastica de bază pot avea un efect benefic asupra asimetriei toracelui, adică diminuarea acesteia. Recomandăm utilizarea acestor exerciții în tratarea persoanelor cu asimetria moderată sau severă a toracelui. Cuvinte cheie: asimetria trunchiului, scolioza, adolescenta, program de activitate fizică.

A szerző áttekintést nyújt a hamisítások technikai és környezeti változásairól az elmúlt két évtizedben.

Accumulation of damaged or misfolded proteins resulted from oxidative protein modification induces endoplasmic reticulum (ER) stress by activating the pathways of unfolded protein response. In pathologic hemolytic conditions, extracellular free hemoglobin is submitted to rapid oxidation causing heme release. Resident cells of atherosclerotic lesions, after intraplaque hemorrhage, are exposed to heme leading to oxidative injury. Therefore, we raised the question whether heme can also provoke ER stress. Smooth muscle cells are one of the key players of atherogenesis; thus, human aortic smooth muscle cells (HAoSMCs) were selected as a model cell to reveal the possible link between heme and ER stress. Using immunoblotting, quantitative polymerase chain reaction and immunocytochemistry, we quantitated the markers of ER stress. These were: phosphorylated eIF2α, Activating transcription factor-4 (ATF4), DNA-damage-inducible transcript 3 (also known as C/EBP homology protein, termed CHOP), X-box binding protein-1 (XBP1), Activating transcription factor-6 (ATF6), GRP78 (glucose-regulated protein, 78kDa) and heme responsive genes heme oxygenase-1 and ferritin. In addition, immunohistochemistry was performed on human carotid artery specimens from patients who had undergone carotid endarterectomy. We demonstrate that heme increases the phosphorylation of eiF2α in HAoSMCs and the expression of ATF4. Heme also enhances the splicing of XBP1 and the proteolytic cleavage of ATF6. Consequently, there is up-regulation of target genes increasing both mRNA and protein levels of CHOP and GRP78. However, TGFβ and collagen type I decreased. When the heme binding proteins, alpha-1-microglobulin (A1M) and hemopexin (Hpx) are present in cell media, the ER stress provoked by heme is inhibited. ER stress pathways are also retarded by the antioxidant N-acetyl cysteine (NAC) indicating that reactive oxygen species are involved in heme-induced ER stress. Consistent with these findings, elevated expression of the ER stress marker GRP78 and CHOP were observed in smooth muscle cells of complicated lesions with hemorrhage compared to either atheromas or healthy arteries. In conclusion, heme triggers ER stress in a time- and dose-dependent manner in HAoSMCs. A1M and Hpx as well as NAC effectively hamper heme-induced ER stress, supporting their use as a potential therapeutic approach to reverse such a deleterious effects of heme toxicity.

Heme oxygenase-1 (HO-1) plays a vital role in the catabolism of heme and yields equimolar amounts of biliverdin, carbon monoxide, and free iron. We report that macrophages engulfing either the low amount of heme-containing apoptotic thymocytes or the high amount of heme-containing eryptotic red blood cells (eRBCs) strongly upregulate HO-1. The induction by apoptotic thymocytes is dependent on soluble signals, which do not include adenylate cyclase activators but induce the p38 mitogen-activated protein (MAP) kinase pathway, while in the case of eRBCs, it is cell uptake-dependent. Both pathways might involve the regulation of BTB and CNC homology 1 (BACH1), which is the repressor transcription regulator factor of the HO-1 gene. Long-term continuous efferocytosis of apoptotic thymocytes is not affected by the loss of HO-1, but that of eRBCs is inhibited. This latter is related to an internal signaling pathway that prevents the efferocytosis-induced increase in Rac1 activity. While the uptake of apoptotic cells suppressed the basal pro-inflammatory cytokine production in wild-type macrophages, in the absence of HO-1, engulfing macrophages produced enhanced amounts of pro-inflammatory cytokines. Our data demonstrate that HO-1 is required for both the engulfment and the anti-inflammatory response parts of the efferocytosis program.

Over the past decades, substantial work has established that hemoglobin oxidation and heme release play a pivotal role in hemolytic/hemorrhagic disorders. Recent reports have shown that oxidized hemoglobins, globin-derived peptides, and heme trigger diverse biological responses, such as toll-like receptor 4 activation with inflammatory response, reprogramming of cellular metabolism, differentiation, stress, and even death. Here, we discuss these cellular responses with particular focus on their mechanisms that are linked to the pathological consequences of hemorrhage and hemolysis. In recent years, endogenous gasotransmitters, such as carbon monoxide (CO) and hydrogen sulfide (H2S), have gained a lot of interest in connection with various human pathologies. Thus, many CO and H2S-releasing molecules have been developed and applied in various human disorders, including hemolytic and hemorrhagic diseases. Here, we discuss our current understanding of oxidized hemoglobin and heme-induced cell and tissue damage with particular focus on inflammation, cellular metabolism and differentiation, and endoplasmic reticulum stress in hemolytic/hemorrhagic human diseases, and the potential beneficial role of CO and H2S in these pathologies. More detailed mechanistic insights into the complex pathology of hemolytic/hemorrhagic diseases through heme oxygenase-1/CO as well as H2S pathways would reveal new therapeutic approaches that can be exploited for clinical benefit.

Hemostasis disorder in patients with end-stage renal disease (ESRD) is frequently associated with bleeding diathesis but it may also manifest in thrombotic complications. Analysis of individual coagulation and fibrinolytic factors may shed light on the background of this paradox situation. Here we explored components essential for fibrin formation/stabilization in ESRD patients being on maintenance hemodiafiltration (HDF) or hemodialysis (HD). Pre-dialysis fibrinogen, factor XIII (FXIII) antigen concentrations and FXIII activity were elevated, while α2-plasmin inhibitor (α2PI) activity decreased. The inflammatory status, as characterized by C-reactive protein (CRP) was a key determinant of fibrinogen concentration, but not of FXIII and α2PI levels. During a 4-h course of HDF or HD, fibrinogen concentration and FXIII levels gradually elevated. When compensated for the change in plasma water, i.e., normalized for plasma albumin concentration, only FXIII elevation remained significant. There was no difference between HDF and HD treatments. Individual HDF treatment did not influence α2PI activity, however after normalization it decreased significantly. HD treatment had a different effect, α2PI activities became elevated but the elevation disappeared after normalization. Elevated fibrinogen and FXIII levels in ESRD patients might contribute to the increased thrombosis risk, while decreased α2PI activity might be associated with elevated fibrinolytic potential.

Heme released from red blood cells targets a number of cell components including the cytoskeleton. The purpose of the present study was to determine the impact of free heme (20–300 µM) on human skeletal muscle fibres made available during orthopedic surgery. Isometric force production and oxidative protein modifications were monitored in permeabilized skeletal muscle fibre segments. A single heme exposure (20 µM) to muscle fibres decreased Ca2+-activated maximal (active) force (Fo) by about 50% and evoked an approximately 3-fold increase in Ca2+-independent (passive) force (Fpassive). Oxidation of sulfhydryl (SH) groups was detected in structural proteins (e.g., nebulin, α-actinin, meromyosin 2) and in contractile proteins (e.g., myosin heavy chain and myosin-binding protein C) as well as in titin in the presence of 300 µM heme. This SH oxidation was not reversed by dithiothreitol (50 mM). Sulfenic acid (SOH) formation was also detected in the structural proteins (nebulin, α-actinin, meromyosin). Heme effects on SH oxidation and SOH formation were prevented by hemopexin (Hpx) and α1-microglobulin (A1M). These data suggest that free heme has a significant impact on human skeletal muscle fibres, whereby oxidative alterations in structural and contractile proteins limit contractile function. This may explain and or contribute to the weakness and increase of skeletal muscle stiffness in chronic heart failure, rhabdomyolysis, and other hemolytic diseases. Therefore, therapeutic use of Hpx and A1M supplementation might be effective in preventing heme-induced skeletal muscle alterations.

Hemoglobin, heme and iron are implicated in the progression of atherosclerosis. Therefore, we investigated whether the hydrophobic fungal iron chelator siderophore, desferricoprogen (DFC) inhibits atherosclerosis. DFC reduced atherosclerotic plaque formation in ApoE−/− mice on an atherogenic diet. It lowered the plasma level of oxidized LDL (oxLDL) and inhibited lipid peroxidation in aortic roots. The elevated collagen/elastin content and enhanced expression of adhesion molecule VCAM-1 were decreased. DFC diminished oxidation of Low-density Lipoprotein (LDL) and plaque lipids catalyzed by heme or hemoglobin. Formation of foam cells, uptake of oxLDL by macrophages, upregulation of CD36 and increased expression of TNF-α were reduced by DFC in macrophages. TNF-triggered endothelial cell activation (vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecules (ICAMs), E-selectin) and increased adhesion of monocytes to endothelium were attenuated. The increased endothelial permeability and intracellular gap formation provoked by TNF-α was also prevented by DFC. DFC acted as a cytoprotectant in endothelial cells and macrophages challenged with a lethal dose of oxLDL and lowered the expression of stress-responsive heme oxygenase-1 as sublethal dose was employed. Saturation of desferrisiderophore with iron led to the loss of the beneficial effects. We demonstrated that DFC accumulated within the atheromas of the aorta in ApoE−/− mice. DFC represents a novel therapeutic approach to control the progression of atherosclerosis.

We previously found higher level of endothelial cell (EC) activation in patients who suffered from in-stent restenosis after bare-metal stenting compared to subjects who underwent drug-eluting stenting (DES) showing no complications. Here we investigated the potential transcriptional and post-transcriptional regulatory mechanisms by which everolimus attenuated EC activation after DES. We studied the effect of everolimus on E-selectin (SELE) and VCAM1 mRNA levels when human coronary artery (HCAECs) and human umbilical vein ECs were challenged with recombinant TNF-α (100 ng/mL) for 1-24 hours in the presence or absence of everolimus using 0.5 μM concentration locally maintained by DES. EC activation was evaluated via the levels of IL-1β and IL-6 mRNAs with miR-155 expression by RT-qPCR as well as the nuclear translocation of nuclear factor kappa beta (NF-κB) detected by fluorescence microscopy. To investigate the transcriptional regulation of E-selectin and VCAM-1, TNF-α-induced enhancer RNA (eRNA) expression at p65-bound enhancers in the neighboring genomic regions of SELE and VCAM1 genes, including SELE_-11Kb and VCAM1_-10Kb, were measured in HCAECs. Mature and precursor levels of E-selectin and VCAM-1 repressor miR-181b were quantified to analyze the post-transcriptional regulation of these genes in HCAECs. Circulating miR-181b was analyzed in plasma samples of stented subjects by stem-loop RT-qPCR. TNF-α highly elevated E-selectin and VCAM-1 expression at transcriptional level in ECs. Levels of mature, pre- and pri-miR-181b were repressed in ECs by TNF-α, while everolimus acted as a negative regulator of EC activation via inhibited translocation of NF-κB p65 subunit into cell nuclei, lowered eRNA expression at SELE and VCAM1 genes-associated enhancers and modulated expression of their post-transcriptional repressor miR-181b. Significant negative correlation was observed between plasma miR-181b and soluble E-selectin and VCAM-1 in patients. In conclusion, everolimus attenuates EC activation via reduced NF-κB p65 translocation causing decreased E-selectin and VCAM-1 expression at transcriptional and post-transcriptional level after DES.

Background. Toxic shock syndrome is a potentially fatal toxin-mediated disease. The role of toxins in this clinical entity made us hypothesize that extracorporeal blood purification with CytoSorb® could play a beneficial role in the clinical management of toxic shock syndrome. This case report describes the successful treatment of toxic shock syndrome using a combination of renal replacement therapy and hemadsorption in a pediatric patient. Case Presentation. A 5-year-old girl with Down’s syndrome presented with an inflamed area surrounding an insect bite, signs of systemic inflammation, and multiple organ failure. As previous attempts of immune modulation therapy were unsuccessful, renal replacement therapy was supplemented by the cytokine absorber CytoSorb. Treatment using this combination was associated with a rapid and significant stabilization in the hemodynamic situation and a decrease in inflammatory mediators within hours after the initiation of therapy. The application of CytoSorb therapy was simple and safe. Conclusion. The use of extracorporeal blood purification with CytoSorb proved potentially beneficial by removing toxins and inflammatory mediators in this case and could therefore play a role in the clinical management of toxic shock syndrome. Whether CytoSorb has the potential to even positively influence mortality in patients with toxic shock syndrome still needs to be confirmed.

The prevalence of vascular disorders continues to rise worldwide. Parallel with that, new pathophysiological pathways have been discovered, providing possible remedies for prevention and therapy in vascular diseases. Growing evidence suggests that endoplasmic reticulum (ER) stress is involved in a number of vasculopathies, including atherosclerosis, vascular brain events, and diabetes. Heme, which is released from hemoglobin or other heme proteins, triggers various pathophysiological consequence, including heme stress as well as ER stress. The potentially toxic free heme is converted by heme oxygenases (HOs) into carbon monoxide (CO), iron, and biliverdin (BV), the latter of which is reduced to bilirubin (BR). Redox-active iron is oxidized and stored by ferritin, an iron sequestering protein which exhibits ferroxidase activity. In recent years, CO, BV, and BR have been shown to control cellular processes such as inflammation, apoptosis, and antioxidant defense. This review covers our current knowledge about how heme induced endoplasmic reticulum stress (HIERS) participates in the pathogenesis of vascular disorders and highlights recent discoveries in the molecular mechanisms of HO-mediated cytoprotection in heme stress and ER stress, as well as crosstalk between ER stress and HO-1. Furthermore, we focus on the translational potential of HIERS and heme oxygenase-1 (HO-1) in atherosclerosis, diabetes mellitus, and brain hemorrhage.

Iron is at the forefront of a number of pivotal biological processes due to its ability to readily accept and donate electrons. However, this property may also catalyze the generation of free radicals with ensuing cellular and tissue toxicity. Accordingly, throughout evolution numerous pathways and proteins have evolved to minimize the potential hazardous effects of iron cations and yet allow for readily available iron cations in a wide variety of fundamental metabolic processes. One of the extensively studied proteins in the context of systemic and cellular iron metabolisms is ferritin. While clinicians utilize serum ferritin to monitor body iron stores and inflammation, it is important to note that the vast majority of ferritin is located intracellularly. Intracellular ferritin is made of two different subunits (heavy and light chain) and plays an imperative role as a safe iron depot. In the past couple of decades our understanding of ferritin biology has remarkably improved. Additionally, a significant body of evidence has emerged describing the significance of the kidney in iron trafficking and homeostasis. Here, we briefly discuss some of the most important findings that relate to the role of iron and ferritin heavy chain in the context of kidney-related diseases and, in particular, vascular calcification, which is a frequent complication of chronic kidney disease.