The midblastula transition defines the onset of Y RNA-dependent DNA replication in Xenopus laevis

Noncoding Y RNAs are essential for the initiation of chromosomal DNA replication in mammalian cell extracts, but their role in this process during early vertebrate development is unknown. Here, we use antisense morpholino nucleotides (MOs) to investigate Y RNA function in Xenopus laevis and zebrafish embryos. We show that embryos in which Y RNA function is inhibited by MOs develop normally until the midblastula transition (MBT) but then fail to replicate their DNA and die before gastrulation. Consistent with this observation, Y RNA function is not required for DNA replication in Xenopus egg extracts but is required for replication in a post-MBT cell line. Y RNAs do not bind chromatin in karyomeres before MBT, but they associate with interphase nuclei after MBT in an origin recognition complex (ORC)-dependent manner. Y RNA-specific MOs inhibit the association of Y RNAs with ORC, Cdt1, and HMGA1a proteins, suggesting that these molecular associations are essential for Y RNA function in DNA replication. The MBT is thus a transition point between Y RNA-independent and Y RNA-dependent control of vertebrate DNA replication. Our data suggest that in vertebrates Y RNAs function as a developmentally regulated layer of control over the evolutionarily conserved eukaryotic DNA replication machinery.     

Fat Storage-inducing Transmembrane Protein 2 Is Required for Normal Fat Storage in Adipose Tissue

Triglycerides within the cytosol of cells are stored in a phylogenetically conserved organelle called the lipid droplet (LD). LDs can be formed at the endoplasmic reticulum, but mechanisms that regulate the formation of LDs are incompletely understood. Adipose tissue has a high capacity to form lipid droplets and store triglycerides. Fat storage-inducing transmembrane protein 2 (FITM2/FIT2) is highly expressed in adipocytes, and data indicate that FIT2 has an important role in the formation of LDs in cells, but whether FIT2 has a physiological role in triglyceride storage in adipose tissue remains unproven. Here we show that adipose-specific deficiency of FIT2 (AF2KO) in mice results in progressive lipodystrophy of white adipose depots and metabolic dysfunction. In contrast, interscapular brown adipose tissue of AF2KO mice accumulated few but large LDs without changes in cellular triglyceride levels. High fat feeding of AF2KO mice or AF2KO mice on the genetically obese ob/ob background accelerated the onset of lipodystrophy. At the cellular level, primary adipocyte precursors of white and brown adipose tissue differentiated in vitro produced fewer but larger LDs without changes in total cellular triglyceride or triglyceride biosynthesis. These data support the conclusion that FIT2 plays an essential, physiological role in fat storage in vivo.     

Phenol utilization by filamentous yeast Trichosporon cutaneum

The investigated strain Trichosporon cutaneum shows well expressed capability for metabolizing high concentrations of phenol, up to 1 g/l, utilizing it as the sole carbon source for the growth and development of the population. The data reported, prove the good perspectives for its application in protecting the environment from phenol pollution. No data about modelling the process of cultivation of Trichosporon cutaneum in phenol media is available in scientific literature up to now. The mathematical model, reported here, consists of two nonlinear differential equations, describing cell growth and substrate consumption. The unknown parameters are estimated following the method of Hooke and Jeeves. A number of simulation investigations are carried out. They prove the adequacy of the model and its applicability in further studies on the processes of growth and phenol uptake of Trichosporon cutaneum.     

Role of hydrolysis conditions in the enzyme-mediated release of reducing sugars from sulphite pulp

Summary The release of reducing sugars from sulphite pulp treated with an Aureobasidium pullulans xylanase preparation was studied. Hydrolysis conditions such as pH, buffer, temperature and time of enzyme treatment were optimized in terms of their effect on sulphite pulp for production of dissolving pulp with an improved quality. Enzyme hydrolysis of pulp was found to be most effective at a temperature of 55°C and at pH 4.7 using sodium acetate buffer.     

A cellular automaton model for the migration of glioma cells

We present a study of in vitro cell migration in two dimensions as a first step towards understanding the mechanisms governing the motility of glioma cells. Our study is based on a cellular automaton model which aims at reproducing the kinetics of a lump of glioma cells deposited on a substrate of collagen. The dynamical effects of cell attraction and motion inertia are introduced through adequate automaton rules. We compare the density profiles given by the model to those obtained experimentally. The result of the best fit indicates a substantial cell-cell attraction due to cell-cell communication through gap junctions (or chemotaxis) and negligible inertia effects during migration. Tracking of individual migrating cells indicates highly convoluted cell trajectories.     

Short-term effect of elevated CO2 concentration and high irradiance on the antioxidant enzymes in bean plants

The effect of short-term exposure to elevated CO₂ concentration and high irradiance on the activity of superoxide dismutase (SOD), ascorbate peroxidase (APX), guaiacol peroxidases (GPX) and catalase (CAT), and on the extent of the lipid peroxidation was studied in bean (Phaseolus vulgaris L.) plants. Plants were exposed for 4 d (8 h a day) to irradiance of 100 (LI) or 1000 (HI) μmol m⁻² s⁻¹ at ambient (CA, 350 μmol mol⁻¹) or elevated (CE, 1300 μmol mol⁻¹) CO₂ concentration. Four-day exposure to CE increased the leaf dry mass in HI plants and RuBPC activity and chlorophyll content in LI plants. Total soluble protein content, leaf dry matter and RuBPC activity were higher in HI than in LI plants, although the HI and CE increased the contents of malonyldialdehyde and H₂O₂. Under CA, exposure to HI increased the activity of APX and decreased the total SOD activity. Under CE, HI treatment also activated APX and led to reduction of both, SOD and GPX, enzymes activities. CE considerably reduced the CAT activity at both irradiances, possibly due to suppressed rate of photorespiration under CE conditions.     

Proliferating cell nuclear antigen (PCNA) as a proliferative marker during embryonic and adult zebrafish hematopoiesis

We investigated the expression of proliferative cell nuclear antigen (PCNA) in zebrafish to delineate the proliferative hematopoietic component during adult and embryonic hematopoiesis. Immunostaining for PCNA and enhanced green fluorescence protein (eGFP) was performed in wild-type and fli1-eGFP (endothelial marker) and gata1-eGFP (erythroid cell marker) transgenic fish. Expression of PCNA mRNA was examined in wild-type and chordin morphant embryos. In adult zebrafish kidney, the renal tubules are surrounded by endothelial cells and it is separated into hematopoietic and excretory compartments. PCNA was expressed in hematopoietic progenitor cells but not in mature neutrophils, eosinophils or erythroid cells. Some PCNA+ cells are scattered in the hematopoietic compartment of the kidney while others are closely associated with renal tubular cells. PCNA was also expressed in spermatogonial stem cells and intestine crypts, consistent with its role in cell proliferation and DNA synthesis. In embryos, PCNA is expressed in the brain, spinal cord and intermediate cell mass (ICM) at 24 h-post fertilization. In chordin morphants, PCNA is significantly upregulated in the expanded ICM. Therefore, PCNA can be used to mark cell proliferation in zebrafish hematopoietic tissues and to identify a population of progenitor cells whose significance would have to be further investigated.     

Prevalence of the Metabolic Syndrome in a Bulgarian Female Population Referred for Bone Density Testing

Objective: To investigate the prevalence of the metabolic syndrome in Bulgarian women referred for bone density screening. Research Methods and Procedures: This was a cross‐sectional clinical study. Subjects were 444 consecutive 30‐ to 75‐year‐old Bulgarian women recruited from the outpatients referred for bone density testing (mean age, 52.67 ± 15.19 years; mean BMI, 26.10 ± 5.71 kg/m²). Height (centimeters), weight (kilograms), and blood pressure were measured. BMI and waist‐to‐hip ratio were calculated. Fasting plasma glucose, blood lipids, and immunoreactive insulinemia (Bayer Corp.‐Diagnostics Div., Tarrytown, NY) were determined. Body composition was analyzed by bioimpedance on a leg‐to‐leg analyser (Tanita TBF‐215; Tanita Corporation, Tokyo, Japan). Results: Of all women, 56.76% had a BMI > 25 kg/m², 45.95% had a waist circumference > 88 cm, and 64.64% had a waist‐to‐hip ratio > 0.8; 59.90% had hypertension; 4.05% had fasting plasma glucose > 7.0 mM, and 42.79% had fasting morning immunoreactive insulinemia = 16 UI/liter; 23.65% had hypercholesterolemia; and 26.35% had hypertriglyceridemia. The prevalence of the metabolic syndrome in this sample, as defined by the National Cholesterol and Education Program‐Adult Treatment Panel III, was 34.91%, and by the modified World Health Organization definition was 37.16%. Discussion: We concluded that Bulgarian women 30 to 75 years old referred for bone density testing have a high prevalence of the metabolic syndrome. Therefore, large‐scale prevention programs are needed in this field.     

Electrically induced protoplast fusion using pulse electric fields for dielectrophoresis

The formation of protoplast chains in suspensions of isolated pea (Pisum sativum cv Ran 1) mesophyll protoplasts induced by electric fields was studied. The chain formation induced by a sine-wave field (2 V, peak to peak; 500-0.1 kHz) was compared to that induced by an alternating pulse field (1 V, amplitude; 0.1-0.4 kHz). An increased number of dielectrophoretically paired protoplasts, formation of protoplast chains in the presence of CaCl₂ up to 5 mm, and protoplast fusion in the presence of 3 mm CaCl₂ were found when the pulse field was applied. The present results suggest the possibility of electrically induced protoplast fusion at cation concentrations that prevent fusion when sine-wave fields are applied.