Lipid peroxidation and biochemical parameters in umbilical cord blood of well-adapted term newborns

Lipid peroxidation (LPX) can play an important role in the development of pathological changes of foetal and neonatal tissues. We investigated LPX and biochemical parameters in plasma from mixed umbilical cord (m.u.c.) blood and acid-base balance (ABB) parameters in m.u.c. blood of well-adapted full-term newborns. LPX products were estimated as thiobarbituric acid reacting substances (TBARS) and were expressed by using of malondialdehyde (MDA) as a standard solution. Intensity of LPX was estimated in vitro in m.u.c. blood plasma without and with added LPX activator (125 μM L-ascorbate plus 5 μM FeSO₄) and in the incubated plasma (30 min, 37°C) under both conditions. Actual TBARS (3.51 ± 0.49 nmol/mL) were determined in the non-incubated plasma without the added LPX activator. Approximately twice higher TBARS were found in the incubated plasma without the LPX activator (7.29 ± 2.17 nmol/mL) or with it (8.57 ± 2.20 nmol/mL), as well as in the non-incubated plasma after its addition (7.38 ± 1.98 nmol/mL). All analysed biochemical parameters (Fe, total iron-binding capacity, uric acid, proteins, Mg, Ca, phosphate, glucose, K, Na, Cl, ALT, AST, GMT, CK, LD, HBD, AMS, ALP, ACP) and ABB parameters were within their reference ranges. The actual TBARS levels were found being positively correlated with α-hydroxybutyrate dehydrogenase (HBD) activity and negatively with pO₂. These results suggest that LPX in m.u.c. blood plasma might be activated. This activation could probably depend on extent of hypoxia. TBARS and their formation in vitro could be suitable parameters of LPX in m.u.c. blood.     

Human MSH6 deficiency is associated with impaired antibody maturation

Ig class-switch recombination (Ig-CSR) deficiencies are rare primary immunodeficiencies characterized by defective switched isotype (IgG/IgA/IgE) production. Depending on the molecular defect, defective Ig-CSR may also be associated with impaired somatic hypermutation (SHM) of the Ig V regions. Although the mechanisms underlying Ig-CSR and SHM in humans have been revealed (at least in part) by studying natural mutants, the role of mismatch repair in this process has not been fully elucidated. We studied in vivo and in vitro Ab maturation in eight MSH6-deficient patients. The skewed SHM pattern strongly suggests that MSH6 is involved in the human SHM process. Ig-CSR was found to be partially defective in vivo and markedly impaired in vitro. The resolution of γH2AX foci following irradiation of MSH6-deficient B cell lines was also found to be impaired. These data suggest that in human CSR, MSH6 is involved in both the induction and repair of DNA double-strand breaks in switch regions.     

Proteomic analysis of iron overload in human hepatoma cells

Iron-mediated organ damage is common in patients with iron overload diseases, namely, hereditary hemochromatosis. Massive iron deposition in parenchymal organs, particularly in the liver, causes organ dysfunction, fibrosis, cirrhosis, and also hepatocellular carcinoma. To obtain deeper insight into the poorly understood and complex cellular response to iron overload and consequent oxidative stress, we studied iron overload in liver-derived HepG2 cells. Human hepatoma HepG2 cells were exposed to a high concentration of iron for 3 days, and protein expression changes initiated by the iron overload were studied by two-dimensional electrophoresis and mass spectrometry. From a total of 1,060 spots observed, 21 spots were differentially expressed by iron overload. We identified 19 of them; 11 identified proteins were upregulated, whereas 8 identified proteins showed a decline in response to iron overload. The differentially expressed proteins are involved in iron storage, stress response and protection against oxidative stress, protein folding, energy metabolism, gene expression, cell cycle regulation, and other processes. Many of these molecules have not been previously suggested to be involved in the response to iron overload and the consequent oxidative stress.     

Comparison of an oligo-chip based assay with PCR method to measure the prevalence of tick-borne pathogenic bacteria in central Slovakia

Ticks are well-known vectors for a wide range of pathogenic microorganisms. We examined the presence of Rickettsia spp., Anaplasma spp., Borrelia spp., Coxiella burnetii and Francisella tularensis in Ixodes ricinus ticks collected in central Slovakia using oligo-chip based assay. Rickettsiae were detected in 5.6% of examined ticks. Borreliae and anaplasmae were identified in 2.1% and 2.8% ticks, respectively. All tested samples were negative for presence of Coxiella burnetii and Francisella tularensis. All these results were compared with those obtained by PCR analysis, and a close correlation between them was found. In addition, rickettsiae of spotted fever group (SFG), Anaplasma phagocytophilum and Borrelia burgdorferi sensu lato were found in ticks using genera or species-specific PCR methods. They are circulating in 10 out of 18 studied localities.     

Mechanical competition alters the cellular interpretation of an endogenous genetic program

The intrinsic genetic program of a cell is not sufficient to explain all of the cell’s activities. External mechanical stimuli are increasingly recognized as determinants of cell behavior. In the epithelial folding event that constitutes the beginning of gastrulation in Drosophila, the genetic program of the future mesoderm leads to the establishment of a contractile actomyosin network that triggers apical constriction of cells and thereby tissue folding. However, some cells do not constrict but instead stretch, even though they share the same genetic program as their constricting neighbors. We show here that tissue-wide interactions force these cells to expand even when an otherwise sufficient amount of apical, active actomyosin is present. Models based on contractile forces and linear stress–strain responses do not reproduce experimental observations, but simulations in which cells behave as ductile materials with nonlinear mechanical properties do. Our models show that this behavior is a general emergent property of actomyosin networks in a supracellular context, in accordance with our experimental observations of actin reorganization within stretching cells.