Hidradenitis suppurativa (HS) is a Th1/17-driven inflammatory skin disease of the apocrine gland-rich (AGR) skin regions, where keratinocytes seem to be the crucial drivers of the initial pathogenic ...steps. However, the possible role of permeability barrier alteration in activating keratinocytes during HS development has not been clarified. We compared the major permeability barrier elements of non-lesional HS (HS-NL; n = 10) and lesional HS (HS-L; n = 10) skin with healthy AGR regions (n = 10) via RT-qPCR and immunohistochemistry. Stratum corneum components related to cornified envelope formation, corneocyte desquamation and (corneo)desmosome organization were analyzed along with tight junction molecules and barrier alarmins. The permeability barrier function was also investigated with transepidermal water loss (TEWL) measurements (n = 16). Junction structures were also visualized using confocal microscopy. At the gene level, none of the investigated molecules were significantly altered in HS-NL skin, while 11 molecules changed significantly in HS-L skin versus control. At the protein level, the investigated molecules were similarly expressed in HS-NL and AGR skin. In HS-L skin, only slight changes were detected; however, differences did not show a unidirectional alteration, as KRT1 and KLK5 were detected in decreased levels, and KLK7, KRT6 and DSG1 in increased levels. No significant differences in TEWL or the expression of junction structures were assessed. Our findings suggest that the permeability barrier is not significantly damaged in HS skin and permeability barrier alterations are not the driver factors of keratinocyte activation in this disease.
Hypertension (HTN) is a major risk factor for heart failure. We investigated the influence of HTN on cardiac contraction and relaxation in transgenic renin overexpressing rats (carrying mouse Ren-2 ...renin gene, mRen2, n = 6). Blood pressure (BP) was measured. Cardiac contractility was characterized by echocardiography, cellular force measurements, and biochemical assays were applied to reveal molecular mechanisms. Sprague-Dawley (SD) rats (n = 6) were used as controls. Transgenic rats had higher circulating renin activity and lower cardiac angiotensin-converting enzyme two levels. Systolic BP was elevated in mRen2 rats (235.11 ± 5.32 vs. 127.03 ± 7.56 mmHg in SD, P < 0.05), resulting in increased left ventricular (LV) weight/body weight ratio (4.05 ± 0.09 vs. 2.77 ± 0.08 mg/g in SD, P < 0.05). Transgenic renin expression had no effect on the systolic parameters, such as LV ejection fraction, cardiomyocyte Ca(2+)-activated force, and Ca(2+) sensitivity of force production. In contrast, diastolic dysfunction was observed in mRen2 compared with SD rats: early and late LV diastolic filling ratio (E/A) was lower (1.14 ± 0.04 vs. 1.87 ± 0.08, P < 0.05), LV isovolumetric relaxation time was longer (43.85 ± 0.89 vs. 28.55 ± 1.33 ms, P < 0.05), cardiomyocyte passive tension was higher (1.74 ± 0.06 vs. 1.28 ± 0.18 kN/m(2), P < 0.05), and lung weight/body weight ratio was increased (6.47 ± 0.24 vs. 5.78 ± 0.19 mg/g, P < 0.05), as was left atrial weight/body weight ratio (0.21 ± 0.03 vs. 0.14 ± 0.03 mg/g, P < 0.05). Hyperphosphorylation of titin at Ser-12742 within the PEVK domain and a twofold overexpression of protein kinase C-α in mRen2 rats were detected. Our data suggest a link between the activation of renin-angiotensin-aldosterone system and increased titin-based stiffness through phosphorylation of titin's PEVK element, contributing to diastolic dysfunction.
The aim of our study was to explore the pathophysiologic role of oxidation of hemoglobin (Hb) to ferrylHb in human atherosclerosis.
We observed a severe oxidation of Hb to ferrylHb in complicated ...atherosclerotic lesions of carotid arteries with oxidative changes of the globin moieties, detected previously described oxidation hotspots in Hb (β1Cys93; β1Cys112; β2Cys112) and identified a novel oxidation hotspot (α1Cys104). After producing a monoclonal anti-ferrylHb antibody, ferrylHb was revealed to be localized extracellularly and also internalized by macrophages in the human hemorrhagic complicated lesions. We demonstrated that ferrylHb is taken up
phagocytosis as well as CD163 receptor-mediated endocytosis and then transported to lysosomes involving actin polymerization. Internalization of ferrylHb was accompanied by upregulation of heme oxygenase-1 and H-ferritin and accumulation of iron within lysosomes as a result of heme/iron uptake. Importantly, macrophages exposed to ferrylHb in atherosclerotic plaques exhibited a proinflammatory phenotype, as reflected by elevated levels of IL-1β and TNF-α. To find further signatures of ferrylHb in complicated lesions, we performed RNA-seq analysis on biopsies from patients who underwent endarterectomies. RNA-seq analysis demonstrated that human complicated lesions had a unique transcriptomic profile different from arteries and atheromatous plaques. Pathways affected in complicated lesions included gene changes associated with phosphoinositide 3-kinase (PI3K) signaling, lipid transport, tissue remodeling, and vascularization. Targeted analysis of gene expression associated with calcification, apoptosis, and hemolytic-specific clusters indicated an increase in the severity of complicated lesions compared with atheroma. A 39% overlap in the differential gene expression profiles of human macrophages exposed to ferrylHb and the complicated lesion profiles was uncovered. Among these 547 genes, we found inflammatory, angiogenesis, and iron metabolism gene clusters regulated in macrophages.
We conclude that oxidation of Hb to ferrylHb contributes to the progression of atherosclerosis
polarizing macrophages into a proatherogenic phenotype.
. 35, 917-950.
Podocytes are highly specialized, arborized epithelial cells covering the outer surface of the glomerular tuft in the kidney. Terminally differentiated podocytes are unable to go through cell ...division and hereby they are lacking a key property for regeneration after a toxic injury. Podocytes are long-lived cells but, to date, little is known about the mechanisms that support their stress resistance. Our aim was to investigate whether the well-known morphological changes during podocyte differentiation are accompanied by changes in oxidative resistance in a manner that could support their long-term survival. We used a conditionally immortalized human podocyte cell line to study the morphological and functional changes during differentiation. We followed the differentiation process for 14 days by time-lapse microscopy. During this period nondifferentiated podocytes gradually transformed into large, nonproliferating, frequently multinucleated cells, with enlarged nuclei and opened chromatin structure. We observed that differentiated podocytes were highly resistant to oxidants such as H2O2 and heme when applied separately or in combination, whereas undifferentiated cells were prone to such challenges. Elevated oxidative resistance of differentiated podocytes was associated with increased activities of antioxidant enzymes and H-ferritin expression. Immunohistochemical analysis of normal human kidney specimens revealed that podocytes highly express H-ferritin in vivo as well.
Correlations between obesity and asthma control in children: Hungarian primary care pilot study Introduction: Asthma is often associated with overweight and obesity. The aim of this study was to find ...associations between asthma control, obesity and different levels of physical activity. Methods: Using a questionnaire, 117 asthmatic patients between 6-18 years of age were interviewed. Two groups of children, normal vs. overweight, at different levels of physical activity (sport, school-based proper or light activity and full physical exemption) were compared. Asthma control was evaluated in two groups (controlled vs. non or partially controlled). Results: The asthmatic status was generally controlled in 78%; in 81% and 72% of patients with a normal weight and overweight, respectively. Being overweight was more common among girls than boys (43.2% vs. 30.3%). A positive family history of asthma was revealed in many cases; the prevalence of asthma was found at 63% (P=0.0074) among the parents. Weight was significantly higher when the parents themselves were overweight. A lightened workload in physical education lessons at school doubled the risk of obesity (P=0.25), while full exemption increased it by six times (P=0.06). High bodyweight was found in 37% and 31% of children who had received steroid medication and other treatment, respectively (P=0.57). Conclusion: High bodyweight and physical inactivity worsened the chances of effective asthma treatment, while sport improved it. The rates of physical activity among the surveyed patients were lower than recommended. More focus is needed; paediatricians, school-teachers and parents should pay more attention to the issue when establishing a proper family background for healthier lifestyles. PUBLICATION ABSTRACT
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.
Hydrogen sulfide (H
S) 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 H
S levels in human aortic valves. Lower levels of bioavailable H
S 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 H
S 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 H
S. The expression of mitochondrial enzymes involved in H
S catabolism including sulfide quinone oxidoreductase (SQR), the key enzyme in mitochondrial H
S 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 H
S 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 H
S levels decreased. The benefit was mediated via CSE induction and H
S generation. We conclude that decreased levels of bioavailable H
S in human calcific aortic valves result from an increased H
S metabolism that facilitates the development of CAVD. CSE/H
S represent a pathway that reverses the action of calcifying stimuli.
Abstract only
Hypertension is a major co‐morbidity of diastolic dysfunction. Appearance of systolic dysfunction on top of diastolic dysfunction further increases the risk of cardiac failure and ...mortality. We hypothesized that a hypertensive rat model with diastolic dysfunction can give insight into the pathomechanism of progressing human heart failure.
To this end we compared 15 weeks old male rats (n=9‐9) with hypertension due to a renin transgene (mRen2, 228.9±4.9 mmHg; P<0.05) with geno‐/phentoype controls (SD, 121.3±4.4 mmHg).
Invasive parameters of left ventricular (LV) active relaxation (Tau, mRen2: 16.9±0.8 ms vs. SD: 12.6±0.6 ms; P<0.05) and passive stiffness (slope of EDPVR, mRen2: 0.050±0.004 mmHg/μL vs. SD:0.040±0.002 mmHg/μL; P<0.05) confirmed diastolic dysfunction with preserved ejection fraction (EF, mRen2: 52.5±0.7% vs. SD:53.3±0.7%) in the mRen2 animals.
In contrast, regional echocardiography (n=8‐8) and histology (n=5‐5) showed worse Tei index (0.78±0.04; P<0.05) and relatively higher wall thickness (0.74±0.06; P<0.05) of the LV free wall in mRen2 rats than those of the interventricular septum (0.68±0.02 and 0.54±0.03, respectively).
At the cellular level, both lower Ca
2+
sensitivity (pCa
50
) and higher troponin I (cTnI) phosphorylation of LV cardiomyocytes from the free wall and the septum (n=mean of 3‐3/4‐4 rats) suggest abnormal systolic function in the mRen2 group as compared to the control. LV heterogeneity of mRen2 rats is further supported by lower cardiomyocyte maximal Ca
2+
‐activated force (21.2±1.4 kN/m
2
), higher Ca
2+
sensitivity (5.85±0.01) and phosphorylation of cTnI at Thr‐144 (1.62±0.19; P<0.05) in the free wall than those in the septum (30.0±0.6 kN/m
2
, 5.76±0.02 and 1.00±0.15, respectively).
The local abnormalities of LV contraction on top of global diastolic dysfunction may provide a pathomechanism for the clinical progression with increasing mortality seen in human.
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