EGFR activation differentially affects the inflammatory profiles of female human aortic and coronary artery endothelial cells.

Endothelial cells (EC) are key players in vascular function, homeostasis and inflammation. EC show substantial heterogeneity due to inter-individual variability (e.g. sex-differences) and intra-individual differences as they originate from different organs or vessels. This variability may lead to different responsiveness to external stimuli. Here we compared the responsiveness of female human primary EC from the aorta (HAoEC) and coronary arteries (HCAEC) to Epidermal Growth Factor Receptor (EGFR) activation. EGFR is an important signal integration hub for vascular active substances with physiological and pathophysiological relevance. Our transcriptomic analysis suggested that EGFR activation differentially affects the inflammatory profiles of HAoEC and HCAEC, particularly by inducing a HCAEC-driven leukocyte attraction but a downregulation of adhesion molecule and chemoattractant expression in HAoEC. Experimental assessments of selected inflammation markers were performed to validate these predictions and the results confirmed a dual role of EGFR in these cells: its activation initiated an anti-inflammatory response in HAoEC but a pro-inflammatory one in HCAEC. Our study highlights that, although they are both arterial EC, female HAoEC and HCAEC are distinguishable with regard to the role of EGFR and its involvement in inflammation regulation, what may be relevant for vascular maintenance but also the pathogenesis of endothelial dysfunction.

Hypothesis-generating analysis of the impact of non-damaging metabolic acidosis on the transcriptome of different cell types: Integrated stress response (ISR) modulation as general transcriptomic reaction to non-respiratory acidic stress?

Extracellular pH is an important parameter influencing cell function and fate. Microenvironmental acidosis accompanies different pathological situations, including inflammation, hypoxia and ischemia. Research focussed mainly on acidification of the tumour micromilieu and the possible consequences on proliferation, migration and drug resistance. Much less is known regarding the impact of microenvironmental acidosis on the transcriptome of non-tumour cells, which are exposed to local acidosis during inflammation, hypoxia, ischemia or metabolic derailment. In the present hypothesis-generating study, we investigated the transcriptional impact of extracellular acidosis on five non-tumour cell types of human and rat origin, combining RNA-Sequencing and extensive bioinformatics analyses. For this purpose, cell type-dependent acidosis resiliences and acidosis-induced transcriptional changes within these resilience ranges were determined, using 56 biological samples. The RNA-Sequencing results were used for dual differential-expression analysis (DESeq and edgeR) and, after appropriate homology mapping, Gene Ontology enrichment analysis (g:Profiler), Ingenuity Pathway Analysis (IPA®), as well as functional enrichment analysis for predicted upstream regulators, were performed. Extracellular acidosis led to substantial, yet different, quantitative transcriptional alterations in all five cell types. Our results identify the regulator of the transcriptional activity NCOA5 as the only general acidosis-responsive gene. Although we observed a species- and cell type-dominated response regarding gene expression regulation, Gene Ontology enrichment analysis and upstream regulator analysis predicted a general acidosis response pattern. Indeed, they suggested the regulation of four general acidosis-responsive cellular networks, which comprised the integrated stress response (ISR), TGF-ß signalling, NFE2L2 and TP53. Future studies will have to extend the results of our bioinformatics analyses to cell biological and cell physiological validation experiments, in order to test the refined working hypothesis here.

The mineralocorticoid receptor leads to increased expression of EGFR and T-type calcium channels that support HL-1 cell hypertrophy.

The EGF receptor (EGFR) has been extensively studied in tumor biology and recently a role in cardiovascular pathophysiology was suggested. The mineralocorticoid receptor (MR) is an important effector of the renin-angiotensin-aldosterone-system and elicits pathophysiological effects in the cardiovascular system; however, the underlying molecular mechanisms are unclear. Our aim was to investigate the importance of EGFR for MR-mediated cardiovascular pathophysiology because MR is known to induce EGFR expression. We identified a SNP within the EGFR promoter that modulates MR-induced EGFR expression. In RNA-sequencing and qPCR experiments in heart tissue of EGFR KO and WT mice, changes in EGFR abundance led to differential expression of cardiac ion channels, especially of the T-type calcium channel CACNA1H. Accordingly, CACNA1H expression was increased in WT mice after in vivo MR activation by aldosterone but not in respective EGFR KO mice. Aldosterone- and EGF-responsiveness of CACNA1H expression was confirmed in HL-1 cells by Western blot and by measuring peak current density of T-type calcium channels. Aldosterone-induced CACNA1H protein expression could be abrogated by the EGFR inhibitor AG1478. Furthermore, inhibition of T-type calcium channels with mibefradil or ML218 reduced diameter, volume and BNP levels in HL-1 cells. In conclusion the MR regulates EGFR and CACNA1H expression, which has an effect on HL-1 cell diameter, and the extent of this regulation seems to depend on the SNP-216 (G/T) genotype. This suggests that the EGFR may be an intermediate for MR-mediated cardiovascular changes and that SNP analysis can help identify subgroups of patients that will benefit most from MR antagonists.

Weighted Correlation Network Analysis Reveals CDK2 as a Regulator of a Ubiquitous Environmental Toxin-Induced Cell-Cycle Arrest.

Environmental food contaminants constitute a threat to human health. For instance, the globally spread mycotoxin Ochratoxin A (OTA) contributes to chronic kidney damage by affecting proximal tubule cells via unknown mechanisms. We applied a top-down approach to identify relevant toxicological mechanisms of OTA using RNA-sequencing followed by in-depth bioinformatics analysis and experimental validation. Differential expression analyses revealed that OTA led to the regulation of gene expression in kidney human cell lines, including for genes enriched in cell cycle-related pathways, and OTA-induced gap 1 and 2 (G1 and G2) cell-cycle arrests were observed. Weighted correlation network analysis highlighted cyclin dependent kinase 2 (CDK2) as a putative key regulator of this effect. CDK2 was downregulated by OTA exposure, and its overexpression partially blocked the OTA-induced G1 but not G2 cell-cycle arrest. We, therefore, propose CDK2 as one of the key regulators of the G1 cell-cycle arrest induced by low nanomolar concentrations of OTA.

Long-term follow-up after anterior cruciate ligament reconstruction using a press-fit quadriceps tendon-patellar bone autograft.

BACKGROUND: The use of a quadriceps tendon-patellar bone (QTPB) autograft provides an alternative procedure in primary reconstruction of the anterior cruciate ligament (ACL). Using the press-fit technique for femoral fixation and knotting over a bone bridge as well as additional spongiosa filling for tibial fixation can completely eliminate the need for fixation implants. The objective of this study was to evaluate the long-term clinical, functional and radiological results of this operating method. METHODS: Sixty-nine patients (27 female-42 male) were included in this study. Fifty-seven patients (83%) received a comprehensive follow-up review after an average period of 7.5 years (range: 7-8.7). All other patients were surveyed by telephone. Six patients (9%) suffered a re-rupture of the ACL graft caused by a new related trauma and were therefore excluded from the statistical analysis. RESULTS: Of all patients, 98% were satisfied with the operation. Normal or almost normal results were recorded in the subjective IKDC scores form by 88% of the patients. The Lysholm score demonstrated very good and good results in 83% of the patients. Only 1 patient reported minor complaints in the donor area. Seven (12%) patients developed Cyclops syndrome with limited knee extension. This complication was treated arthroscopically within the first year postoperatively. Their results on follow-up were not worse than the results of the patients without Cyclops syndrome. Regarding the 57 patients who received a comprehensive evaluation, the stability test with the KT-1000 Arthrometer yielded a difference of less than 3 mm in the contralateral comparison for 89% of the operated knees. The pivot-shift test was normal in 79% and almost normal in 21%. In the Single-leg Triple Hop Test, patients achieved an average of 98% of the hopping distance attained with the contralateral leg. The radiological examination revealed a slight deterioration in the Kellgren-Lawrence Score in 2 patients. CONCLUSION: The ACL reconstruction using the QTPB autograft performed with the press-fit technique leads to good results in comparison with published results of established procedures for primary ACL surgery using other autografts. Further investigations should involve comparative studies with the objective of providing evidence-based, individually adapted therapy for ACL rupture.

Using gas mixtures of CO, CO2 and H2 as microbial substrates: the do's and don'ts of successful technology transfer from laboratory to production scale.

The reduction of CO2 emissions is a global effort which is not only supported by the society and politicians but also by the industry. Chemical producers worldwide follow the strategic goal to reduce CO2 emissions by replacing existing fossil-based production routes with sustainable alternatives. The smart use of CO and CO2 /H2 mixtures even allows to produce important chemical building blocks consuming the said gases as substrates in carboxydotrophic fermentations with acetogenic bacteria. However, existing industrial infrastructure and market demands impose constraints on microbes, bioprocesses and products that require careful consideration to ensure technical and economic success. The mini review provides scientific and industrial facets finally to enable the successful implementation of gas fermentation technologies in the industrial scale.

Perioperative Risks of Untreated Obstructive Sleep Apnea in the Bariatric Surgery Patient: a Retrospective Study.

BACKGROUND: The use of continuous positive airway pressure (CPAP) perioperatively in bariatric surgery patients with obstructive sleep apnea (OSA) has been controversial. Although studies have demonstrated that CPAP use is safe in bariatric patients, prior studies have not shown improvement in outcomes in these patients. METHODS: A retrospective review of patients who underwent bariatric surgery from 2005 to 2009 was performed. All patients underwent polysomnography preoperatively. Patient age, sex, BMI, comorbidities, polysomnogram data, type of bariatric procedure, length of hospital stay, and postoperative complications were reviewed. The Fisher exact test was used for statistical analysis. RESULTS: Among the 352 patients studied, 47 with apnea-hypopnea index (AHI) ≥5 did not receive CPAP postoperatively. A total of 7/47 (14.9 %) developed postoperative pulmonary complications. There were no non-pulmonary complications. Some 9/305 (2.95 %) with CPAP developed pulmonary complications. There were 26/305 patients with all-cause complications (8.52 %). The AHI was higher in the group receiving treatment. There was a statistically significant difference in pulmonary complications between patients with and those without treatment (p value 0.0002). The average length of stay was 3.0 and 3.2 days in treatment and comparison groups, respectively, a difference that did not reach statistical significance. CONCLUSIONS: Patients who did not receive CPAP postoperatively developed more pulmonary complications than those with CPAP, suggesting that CPAP might be beneficial in decreasing pulmonary complications in patients undergoing bariatric surgery. However, further investigation is warranted to better delineate other risk factors due to small sample size in our study group.

Collective cell migration induced by mechanical stress and substrate adhesiveness.

Mechanical stress normal to the boundary of a tissue sheet can arise in both constrained as well as unconstrained epithelial layers through pushing and pulling of surrounding tissue and substrate adhesiveness, respectively. A continuum model is used to investigate how such stress influences the epithelial dynamics. Four types of spreading and motility can be identified: a uniformly stretched stationary state, uniform sheet migration, active stress compensation by polarization close to the boundary, and a wormlike progression by deformation waves. Analytical and numerical solutions are presented along with bifurcation diagrams using normal stress and active force as control parameters.

Recent Advancements in Prognostic Factors of Epithelial Ovarian Carcinoma.

Ovarian cancer remains the most common cause of gynecologic cancer-related death among women in developed countries. Nevertheless, subgroups of ovarian cancer patients experience relatively longer survival. Efforts to identify prognostic factors that characterize such patients are ongoing, with investigational areas including tumor characteristics, surgical management, inheritance patterns, immunologic factors, and genomic patterns. This review discusses various demographic, clinical, and molecular factors implicating longevity and ovarian cancer survival. Continued efforts at identifying these prognosticators may result in invaluable adjuncts to the treatment of ovarian cancer, with the ultimate goal of advancing patient care.

Free testosterone drives cancer aggressiveness: evidence from US population studies.

Cancer incidence and mortality are higher in males than in females, suggesting that some gender-related factors are behind such a difference. To analyze this phenomenon the most recent Surveillance, Epidemiology and End Results (SEER) database served to access cancer survival data for the US population. Patients with gender-specific cancer and with limited information were excluded and this fact limited the sample size to 1,194,490 patients. NHANES III provided the distribution of physiologic variables in US population (n = 29,314). Cox model and Kaplan-Meier method were used to test the impact of gender on survival across age, and to calculate the gender-specific hazard ratio of dying from cancer five years following diagnosis. The distribution of the hazard ratio across age was then compared with the distribution of 65 physiological variables assessed in NHANES III. Spearman and Kolmogorov-Smirnov test assessed the homology. Cancer survival was lower in males than in females in the age range 17 to 61 years. The risk of death from cancer in males was about 30% higher than that of females of the same age. This effect was present only in sarcomas and epithelial solid tumors with distant disease and the effect was more prominent in African-Americans than Caucasians. When compared to the variables assessed in the NHANES III study, the hazard ratio almost exactly matched the distribution of free testosterone in males; none of the other analyzed variables exhibited a similar homology. Our findings suggest that male sex hormones give rise to cancer aggressiveness in patients younger than 61 years.

Infective endocarditis in two young men with previously undiagnosed bicuspid aortic valves.

Infective endocarditis (IE) is an uncommon, potentially life-threatening microbial infection of the endocardial surface of the heart. An increased risk for IE is associated with a variety of pre-existing structural intracardiac conditions, including a bicuspid aortic valve, which provide a nidus for infection. A bicuspid aortic valve affects 0.5 to 2% of the population, the majority of patients are male, and patients usuallyremain unaware of their condition until infection or calcific valvular dysfunction occurs. We report two cases of native valve IE in otherwise healthy young men seen within a recent two-month period; both cases involved a previously undiagnosed bicuspid aortic valve and required a combination of aggressive antimicrobial therapy and early surgery as definitive treatment. Any patient, especially a young male, presentingwith a febrile illness and a previously or newly diagnosed heart murmur should raise the index of clinical suspicion for this disease.