Effects of rHuEPO treatment on red blood cell osmotic resistance

Red blood cell osmotic resistance (RBCOR) is defined as resistance to osmotic changes in cell integrity after their exposure to hypotonic saline solution. The investigation examined the effect of rHuEPO on RBCOR in hemodialysed patients. The study included 58 patients aged 49 +/- 14 years, treated by hemodialysis for 59 +/- 43 months on average. Half of the patients received rHuEPO for anemia correction. RBCOR was determined in all patients as 3 values: hemolysis start point (HSP), hemolysis end point (HEP) and middle osmotic resistance (MOR). The patients underwent laboratory checkup for parameters characteristically changed in the uremic syndrome. In the control group of healthy subjects (n = 16) RBCOR was only determined. No differences were found in the average values of HSP, HEP and MOR between the rHuEPO treated group of patinets and the untreated group. Compared to healthy individuals, the hemodialysed patients displayed significantly higher values of HSP, HEP and MOR. The only one significant correlation of RBCOR and routine laboratory features was found between MOR and predialytic serum concentrations of calcium (r = 0.28, p < 0.05) and hydrogen ions (r = 0.37, p < 0.05). Our results suggest that the administration of rHuEPO does not affect RBCOR in hemodialysed patients, that RBCOR is not always reduced in this population and that it correlates with a small number of laboratory parameters characteristic for the uremic syndrome.     

Socio-demographic characteristics and lifestyle habits of pregnant women smokers

The aim of this study was to describe anthropometric, clinical, socio-demographic characteristics and lifestyle habits of pregnant smokers in comparison to pregnant nonsmokers. During years 1999-2003, 1,435 pregnant smokers and 4,772 pregnant nonsmokers were interviewed after delivery with a questionnaire. They were recorded clinical, anthropometric and socio-demographic data, smoking status, labor outcome, maternal and fetal hemoglobin concentrations for each patient. The two groups were comparable in anthropometric and clinical characteristics, duration of pregnancy and mode of delivery, except for birth weights, which were significantly lower in newborns of smokers. Maternal hemoglobin concentrations were significantly lower in smokers, but fetal hemoglobin concentrations were significantly higher in babies of smokers. The proportion of pregnant women who smoked during pregnancy was higher among urban women, among women with lower educational level and among unemployed subjects in comparison with nonsmokers. The pregnant women who smoked during pregnancy were more often caffeine and alcohol consumers. To further reduce smoking during pregnancy it is important to continue to promote smoking cessation among teenagers.     

Comparison of fetal plasma cortisol level between eutrophic and hypotrophic newborns

We tested two groups of singletons born at term: fifty-six eutrophic newborns and 56 hypotrophic subjects. They were selected randomly from all newborns delivered by vaginal route between 8 and 14 hours. Excluded were preeclampsia, diabetes, labours longer than 12 hours and newborns with malformations. Written informed consent was obtained from all women and data were collected before and after labour. Umbilical cord blood samples were obtained immediately following the delivery and plasma cortisol concentrations were measured by radioimmunoassay. The groups did not differ significantly regarding maternal age, parity, gestational age and Apgar score, but birth weight was significant differed (p < 0.001). In addition, eutrophic newborns had significantly elevated cortisol levels (457.7 nmol/L, 321.8-696.6 nmol/L) compared with hypotrophic newborns (320.5 nmol/L, 215.1-578.7 nmol/L, p < 0.001). The role of fetal cortisol in intrauterine growth restriction (IUGR) pregnancy and labour is uncertain, but fetal plasma cortisol levels may be lower in IUGR newborns.     

Biodiversity of cryopegs in permafrost

This study describes the biodiversity of the indigenous microbial community in the sodium-chloride water brines (cryopegs) derived from ancient marine sediments and sandwiched within permafrost 100-120,000 years ago after the Arctic Ocean regression. Cryopegs remain liquid at the in situ temperature of -9 to -11 degrees C and make up the only habitat on the Earth that is characterized by permanently subzero temperatures, high salinity, and the absence of external influence during geological time. From these cryopegs, anaerobic and aerobic, spore-less and spore-forming, halotolerant and halophilic, psychrophilic and psychrotrophic bacteria, mycelial fungi and yeast were isolated and their activity was detected below 0 degrees C.     

Environmental and Hormonal Physiology of Postharvest Needle Abscission in Christmas Trees

Several of the conifer species are increasingly adopted as Christmas trees worldwide. These species have become integral parts of the horticultural economies of North American and European countries. Postharvest characteristics such as needle abscission/retention, color, fragrance and rehydration abilities vary with species and these complex physiological traits are strongly modulated by hormonal and environmental factors. A large body of research indicates that prevalence of low temperature before harvest evokes cold acclimation responses that involve an increase in complex sugar concentrations, alterations in membrane structures and enhancements in scavenging abilities promoting postharvest needle retention. Adverse postharvest environmental factors, for example, high temperature and vapor pressure deficit are found to increase water stress, cause dehydration and accelerate needle abscission and/or discoloration. Postharvest water stress/cellular dehydration is one of the fundamental biophysical signals that triggers a cascade of hormonal changes, leading to needle abscission. Abscissic acid levels increase during cold acclimation as well as prior to abscission indicating a complicated and paradoxical role in abscission. Ethylene levels increase before abscission and are well proven to instigate the needle fall. Concentrations of cytokinins, auxins and polyamines decline postharvest. However, their interactive roles with other phytohormones orchestrating the abscission process still remain elusive. This review presents and discusses our current knowledge of the physiological aspects of pre-and postharvest environmental factors on needle abscission.     

Mitochondrial genomes of the genus Ephydatia Lamouroux, 1816: can palindromic elements be used in species-level studies?

Poriferan mitochondrial DNA (mtDNA), especially large intergenic regions, is a target for the insertion of repetitive hairpin-forming elements. These elements are responsible for the large mt genome size differences observed even among closely related sponge taxa. In this study, we present the new, nearly complete, mt genome sequence of Ephydatia fluviatilis and compare it with previously published mt genomes of freshwater sponges. Special emphasis was placed on comparison with the closely related species Ephydatia muelleri, thereby comparing the only two species of the genus Ephydatia on the western Balkan Peninsula. In particular, we analyzed repetitive palindromic elements within the mitochondrial intergenic regions. The genomic distribution of these repetitive elements was analyzed and their potential role in the evolution of mt genomes discussed. We show here that palindromic elements are widespread through the whole mt genome, including the protein coding genes, thus introducing genetic variability into mt genomes.     

Compensation for the absence of the catalytically active half of DNA polymerase ε in yeast by positively selected mutations in CDC28

Current eukaryotic replication models postulate that leading and lagging DNA strands are replicated predominantly by dedicated DNA polymerases. The catalytic subunit of the leading strand DNA polymerase ε, Pol2, consists of two halves made of two different ancestral B-family DNA polymerases. Counterintuitively, the catalytically active N-terminal half is dispensable, while the inactive C-terminal part is required for viability. Despite extensive studies of yeast Saccharomyces cerevisiae strains lacking the active N-terminal half, it is still unclear how these strains survive and recover. We designed a robust method for constructing mutants with only the C-terminal part of Pol2. Strains without the active polymerase part show severe growth defects, sensitivity to replication inhibitors, chromosomal instability, and elevated spontaneous mutagenesis. Intriguingly, the slow-growing mutant strains rapidly accumulate fast-growing clones. Analysis of genomic DNA sequences of these clones revealed that the adaptation to the loss of the catalytic N-terminal part of Pol2 occurs by a positive selection of mutants with improved growth. Elevated mutation rates help generate sufficient numbers of these variants. Single nucleotide changes in the cell cycle-dependent kinase gene, CDC28, improve the growth of strains lacking the N-terminal part of Pol2, and rescue their sensitivity to replication inhibitors and, in parallel, lower mutation rates. Our study predicts that changes in mammalian homologs of cyclin-dependent kinases may contribute to cellular responses to the leading strand polymerase defects.     

Mislocalization of CDK11/PITSLRE, a regulator of the G2/M phase of the cell cycle, in Alzheimer disease

Post-mitotic neurons are typically terminally differentiated and in a quiescent status. However, in Alzheimer disease (AD), many neurons display ectopic re-expression of cell cycle-related proteins. Cyclin-dependent kinase 11 (CDK11) mRNA produces a 110-kDa protein (CDK11(p110)) throughout the cell cycle, a 58-kDa protein (CDK11(p58)) that is specifically translated from an internal ribosome entry site and expressed only in the G(2)/M phase of the cell cycle, and a 46-kDa protein (CDK11(p46)) that is considered to be apoptosis specific. CDK11 is required for sister chromatid cohesion and the completion of mitosis. In this study, we found that the expression patterns of CDK11 vary such that cytoplasmic CDK11 is increased in AD cellular processes, compared to a pronounced nuclear expression pattern in most controls. We also investigated the effect of amyloid precursor protein (APP) on CDK11 expression in vitro by using M17 cells overexpressing wild-type APP and APP Swedish mutant phenotype and found increased CDK11 expression compared to empty vector. In addition, amyloid-β(25-35) resulted in increased CDK11 in M17 cells. These data suggest that CDK11 may play a vital role in cell cycle re-entry in AD neurons in an APP-dependent manner, thus presenting an intriguing novel function of the APP signaling pathway in AD.     

The uptake of persistent organic pollutants by plants

In a field experiment, the transfer of polychlorinated biphenyls (PCBs) and polycyclic aromatic hydrocarbons (PAHs) from contaminated soil to maize (Zea mays L.), sunflower (Helianthus annuus), poplar (Populus nigra × P. maximowiczii) and willow (Salix × smithiana) and the distribution of PCB congeners in maize and sunflower was investigated. The former waste incinerator in Hradec Králové (Czech Republic) was chosen for the experiment. Results of plot screening showed heterogenous contamination by PCBs and PAHs. PCB soil contamination was evidently caused by Delor 106 or Aroclor 1260 stocking and PAH contamination by chemicals containing fluoranthene, benzo/b/fluoranthene, phenanthrene and pyrene. Tested plants were planted on a contaminated field site, in soil contaminated with 1530 μg/kg of total PCBs and 0.138 and 3.42 mg/kg of total PAHs. The results show that maize and sunflower roots accumulated the most PCBs from soil. These plants accumulated hexa- and heptachlorobiphenyl congeners more than tri-, tetra-, and pentachlorobiphenyl congeners. Total concentrations of PAHs in tested plants ranged from 0.096 to 1.34 mg/kg. The highest phenanthrene concentration was found in aboveground biomass of sunflower and the highest concentration of pyrene, in maize roots.     

Replication protein A (RPA) hampers the processive action of APOBEC3G cytosine deaminase on single-stranded DNA

BACKGROUND: Editing deaminases have a pivotal role in cellular physiology. A notable member of this superfamily, APOBEC3G (A3G), restricts retroviruses, and Activation Induced Deaminase (AID) generates antibody diversity by localized deamination of cytosines in DNA. Unconstrained deaminase activity can cause genome-wide mutagenesis and cancer. The mechanisms that protect the genomic DNA from the undesired action of deaminases are unknown. Using the in vitro deamination assays and expression of A3G in yeast, we show that replication protein A (RPA), the eukaryotic single-stranded DNA (ssDNA) binding protein, severely inhibits the deamination activity and processivity of A3G. PRINCIPAL FINDINGS/METHODOLOGY: We found that mutations induced by A3G in the yeast genomic reporter are changes of a single nucleotide. This is unexpected because of the known property of A3G to catalyze multiple deaminations upon one substrate encounter event in vitro. The addition of recombinant RPA to the oligonucleotide deamination assay severely inhibited A3G activity. Additionally, we reveal the inverse correlation between RPA concentration and the number of deaminations induced by A3G in vitro on long ssDNA regions. This resembles the "hit and run" single base substitution events observed in yeast. SIGNIFICANCE: Our data suggest that RPA is a plausible antimutator factor limiting the activity and processivity of editing deaminases in the model yeast system. Because of the similar antagonism of yeast RPA and human RPA with A3G in vitro, we propose that RPA plays a role in the protection of the human genome cell from A3G and other deaminases when they are inadvertently diverged from their natural targets. We propose a model where RPA serves as one of the guardians of the genome that protects ssDNA from the destructive processive activity of deaminases by non-specific steric hindrance.