István Kocsis

Genetic Variants of TNF-α, IL-1β, IL-4 Receptor α-Chain, IL-6 and IL-10 Genes Are Not Risk Factors for Sepsis in Low-Birth-Weight Infants

The amount of inflammatory cytokines is a major determinant for the development of sepsis in very-low-birth-weight (VLBW) neonates. We investigated whether variants of tumor necrosis factor-α, interleukin (IL)-1β, IL-4 receptor α-chain, IL-6 and IL-10 genes, associated with altered cytokine production, might influence the risk and complications of sepsis in VLBW infants. We determined the presence of these genetic variants in dried blood samples of 33 septic, 35 infected and 35 healthy VLBW neonates by PCR and RFLP methods and analyzed their association with the risk and complications of sepsis. The frequencies of genetic variants did not differ in uninfected and in infected infants with or without sepsis. Moreover, none of the studied complications was associated with carrier state of any of genetic variants. Four of the 5 septic neonates with disseminated intravascular coagulation, however, carried simultaneously the variants of IL-1β and IL-10 genes. We concluded that these genetic polymorphisms do not influence the risk and course of sepsis in VLBW neonates.

Biallelic genotype distributions in papers published in Gut between 1998 and 2003: altered conclusions after recalculating the Hardy-Weinberg equilibrium

The Hardy-Weinberg law1–4 presents a mathematical statement that describes the relationship between gene frequencies and genotype frequencies: gene frequencies at a locus in a randomly interbreeding diploid population and population genotype frequencies remain constant from generation to generation if mating is random and mutation, selection, and migration do not occur. The law states a fundamental principle of population genetics that is approximately true for small, and holds with increasing exactness for large populations. Should the frequencies be perturbed for any reason, they will come to the expected equilibrium frequencies after one generation of random mating. The Hardy-Weinberg law can be used for analytical purposes. It is suitable to test the hypothesis of panmixia and evolutionary stasis. Moreover, it represents a null hypothesis to test in genetic studies. However, according to our personal experience, data for Hardy-Weinberg equilibrium (HWE) calculations in studied populations are not always presented in articles with data on the genotype distributions of biallelic polymorphisms of Mendelian inheritance. In this retrospective survey, we tested in papers published in Gut , if this important and qualifying law was checked in studies investigating genetic polymorphisms between 1998 and April 2003. We collected genotype distributions published in papers in Gut from 1998 (volume 42) to 2003 (volume 52). Of 2389 total publications, we found 69 where genetic polymorphisms were part of the study. Of these, those articles that fulfilled the following criteria were selected: investigation of biallelic genetic polymorphism with Mendelian inheritance; use of healthy reference population in the study; and availability of genotype distribution data. We recalculated HWE in each paper and in each study group. For this purpose, we used Arlequin software (http://anthropologie.unige.ch/arlequin/).5,6 The level of statistical significance was set at p<0.05. Deviations from …

Angiotensin II type 1 receptor A1166C polymorphism and prophylactic indomethacin treatment induced ductus arteriosus closure in very low birth weight neonates

Altered pulmonary vascular resistance might be a factor for delayed closure of the ductus arteriosus (DA) in preterm infants. Angiotensin II plays a central role in the elevation of pulmonary vascular resistance. Angiotensin II exerts its vasoconstrictor effect on the angiotensin II type 1 receptor (AT1R). Homozygous carriers of the AT1R A1166C genetic variant present an exaggerated vasoconstrictor response to angiotensin II. We have investigated whether the presence of AT1R CC1166 influences the effect of prophylactic indomethacin treatment on the closure of DA until the fifth postnatal day in preterm infants. In this retrospective study detailed medical history of the first postnatal week was obtained in 159 infants born before the 33rd gestational week. All were treated by prophylactic indomethacin to induce permanent closure of the DA. On the sixth postnatal day the DA was still open in 56, whereas it was permanently closed in 103. The AT1R A1166C genotype of the infants was determined from Guthrie spots. Stepwise binary logistic regression analysis was used to assess the effect of medical conditions and genotype on the risk of patent DA (PDA). Birth weight, infantile respiratory distress, and severe hypotension were independent risk factors for PDA (p < 0.01, p < 0.05, p < 0.05, respectively). The carrier state of AT1R CC1166 was protective against PDA (p < 0.05; odds ratio, 0.067). AT1R AC1166 genotype was not associated with PDA. Our results indicate that the risk of PDA might be lower in infants of AT1R CC1166 than in those with AC or AA genotypes.

Short-term omeprazole treatment does not influence biochemical parameters of bone turnover in children

Gastric proton pump inhibitors are widely used in the treatment of dyspeptic problems and for the eradication of H. pylori infection. Data are not available on whether omeprazole, a representative of proton pump inhibitors, influences the function of osteoclastic H+-pump in children. We studied the impact of short-term omeprazole administration on the biochemical parameters of bone turnover in pediatric patients. Urinary calcium excretion, serum total alkaline phosphatase activity, collagen type 1 crosslinked C-telopeptide, and osteocalcin levels were determined in 34 children [20 girls (9 prepubertal) and 14 boys (6 prepubertal)] before and after 2 weeks of omeprazole treatment at a dose of 20 mg/day. The measured parameters were within the healthy reference range in each patient. None of them altered during the study in any age or in any gender. We conclude that omeprazole, at a dose of 20 mg/day, does not significantly influence the investigated biochemical parameters of osteoclast and osteoblast function in pediatric patients.

Developmental changes in erythrocyte Na+,K+-ATPase subunit abundance and enzyme activity in neonates

AIM To study the relation between erythrocyte Na+,K+-ATPase subunit isoform composition, Na+,K+-ATPase activity, and cation pump function in preterm and term neonates. DESIGN Erythrocyte Na+,K+-ATPase subunit isoform abundance, Na+,K+-ATPase activity, and cation pump function were studied in blood samples obtained from 56 preterm neonates of 28–32 weeks gestation (group 1), 58 preterm neonates of 33–36 weeks gestation (group 2), and 122 term neonates (group 3) during the first two postnatal days. RESULTS α1isoform abundance was higher and β2 isoform abundance was lower in group 1 than in group 3 (p = 0.0002). α2 and β1 isoform abundance did not change with maturation and there was no evidence for the presence of the α3 isoform. Gestational age was inversely related to Na+,K+-ATPase activity (p = 0.0001) and directly related to intracellular Na+ concentration (p = 0.0025). CONCLUSIONS Expression of the α1 and β2Na+,K+-ATPase subunit isoforms is developmentally regulated. The increased abundance of α1isoforms of immature neonates translates to increased ATPase activity. The lower intracellular Na+ concentration of immature neonates suggests that their erythrocyte Na+,K+-ATPase cation pump function may also be increased.

Technical Approach Determines Inflammatory Response after Surgical and Transcatheter Aortic Valve Replacement

Objective To investigate the periprocedural inflammatory response in patients with isolated aortic valve stenosis undergoing surgical aortic valve replacement (SAVR) or transcatheter aortic valve implantation (TAVI) with different technical approaches. Material and Methods Patients were prospectively allocated to one of the following treatments: SAVR using conventional extracorporeal circulation (CECC, n = 47) or minimized extracorporeal circulation (MECC, n = 15), or TAVI using either transapical (TA, n = 15) or transfemoral (TF, n = 24) access. Exclusion criteria included infection, pre-procedural immunosuppressive or antibiotic drug therapy and emergency indications. We investigated interleukin (IL)-6, IL-8, IL-10, human leukocyte antigen (HLA-DR), white blood cell count, high-sensitivity C-reactive protein (hs-CRP) and soluble L-selectin (sCD62L) levels before the procedure and at 4, 24, and 48 h after aortic valve replacement. Data are presented for group interaction (p-values for inter-group comparison) as determined by the Greenhouse-Geisser correction. Results SAVR on CECC was associated with the highest levels of IL-8 and hs-CRP (p<0.017, and 0.007, respectively). SAVR on MECC showed the highest descent in levels of HLA-DR and sCD62L (both p<0.001) in the perioperative period. TA-TAVI showed increased intraprocedural concentration and the highest peak of IL-6 (p = 0.017). Significantly smaller changes in the inflammatory markers were observed in TF-TAVI. Conclusion Surgical and interventional approaches to aortic valve replacement result in inflammatory modulation which differs according to the invasiveness of the procedure. As expected, extracorporeal circulation is associated with the most marked pro-inflammatory activation, whereas TF-TAVI emerges as the approach with the most attenuated inflammatory response. Factors such as the pre-treatment patient condition and the extent of myocardial injury also significantly affect inflammatory biomarker patterns. Accordingly, TA-TAVI is to be classified not as an interventional but a true surgical procedure, with inflammatory biomarker profiles comparable to those found after SAVR. Our study could not establish an obvious link between the extent of the periprocedural inflammatory response and clinical outcome parameters.

Expression and Activity of the Ca2+-ATPase Enzyme in Human Neonatal Erythrocytes

The plasma membrane Ca2+-ATPase (PMCA) is one of the main regulators of Ca2+ homeostasis. We studied the perinatal alteration of the abundance and the activity of PMCA molecules in human erythrocytes in pre-term and full-term neonates and children at the age of 1–4 years. The lower abundance of the 4b isoform was associated with lower enzyme activity in full-term neonates compared to children. Although the number of PMCA molecules was higher in pre-term neonates, their total PMCA activities were identical to those of full-term neonates. Our findings suggest that the abundance of PMCA molecules changes during the perinatal development. The same activity at higher enzyme molecule numbers might indicate a potential immaturity of the enzyme in the pre-term infant.

Angiotensin-converting enzyme DD genotype is preventive against circulatory failure in very-low-birthweight neonates

UNLABELLED We studied the association between genetic polymorphisms of the renin-angiotensin system and the risk for circulatory failure (CF) during the first three postnatal days in 104 very-low-birthweight preterm infants. CONCLUSION Infants with angiotensin-converting enzyme DD genotype were protected against CF (adjusted OR 0.41, 95% CI 0.19-0.89).

CD62L (L-selectin) shedding for assessment of perioperative immune sensitivity in patients undergoing cardiac surgery with cardiopulmonary bypass.

Objective To investigate the suitability of blood granulocyte and monocyte sensitivity, as measured by the quantity of different agonists required to induce CD62L shedding, for assessment of perioperative immune changes in patients undergoing cardiac surgery with cardiopulmonary bypass. Methods Patients scheduled for aortocoronary bypass grafting or for valve surgery were included in this prospective observational study. Blood samples were drawn before anesthesia induction, directly after surgery and 48 hours after anesthesia induction. We determined the concentration of two different inflammatory stimuli – lipoteichoic acid (LTA) and tumor necrosis factor alpha (TNF) - required to induce shedding of 50% of surface CD62L from blood granulocytes and monocytes. In parallel monocyte surface human leukocyte antigen (HLA)-DR, and plasma interleukin (IL)-8, soluble (s)CD62L, soluble (s)Toll-like receptor (TLR)-2 and ADAM17 quantification were used to illustrate perioperative immunomodulation. Results 25 patients were enrolled. Blood granulocytes and monocytes showed decreased sensitivity to the TLR 2/6 agonist Staphylococcus aureus LTA immediately after surgery (p = 0.001 and p = 0.004 respectively). In contrast, granulocytes (p = 0.01), but not monocytes (p = 0.057) displayed a decreased postoperative sensitivity to TNF. We confirmed the presence of a systemic inflammatory response and a decreased immune sensitivity in the post-surgical period by measuring significant increases in the perioperative plasma concentration of IL-8 (p≤0.001) and sTLR (p = 0.004), and decreases in monocyte HLA-DR (p<0.001), plasma sCD62L (p≤0.001). In contrast, ADAM17 plasma levels did not show significant differences over the observation period (p = 0.401). Conclusions Monitoring granulocyte and monocyte sensitivity using the “CD62L shedding assay” in the perioperative period in cardiac surgical patients treated with the use of cardiopulmonary bypass reveals common changes in sensitivity to TLR2/6 ligands and to TNF stimulus. Further long-term follow-up studies will address the predictive value of these observations for clinical purposes.

Determination of H+/K+-ATPase activity in human gastric biopsy specimens.

Abstract The aim of our work was to develop a method to determine the the H+/K+-ATPase activity of human gastric biopsy samples. Our method is based on the phosphatase activity and the K+-induceable property of the enzyme. K+-induceable pNPPase activity was determined from homogenated corpus and antrum biopsy samples. H+/K+-ase activity was calculated as the difference between the corpus and antrum K+-induceable pNPPase activities. Quality control measurements were done during 20 successive days from pooled homogenates. The total, between-day and between-run, within-day and within-run coefficients of variations were between 10 and 16%. The healthy mean and reference range of K+-induceable pNPPase activity in the corpus was 95.8 (95% CI: 83.4–108.2 mU/mg protein); in the antrum it was 28.3 (21.6–35.0) mU/mg protein. The calculated H+/K+-ATPase activity was 67.2 (56.9–77.5) mU/mg protein. The measured activities were independent of the age and gender. Summarizing our results we have concluded, that our novel method might be a potential tool to gather data about the functional acid producing capability of human gastric mucosa.