Immobilization and kinetics of catalase onto magnesium silicate

Bovine liver catalase was immobilized covalently with glutaraldehyde, or glutaraldehyde+3-aminopropionic acid as a spacer, onto magnesium silicate. The coupling time was determined as 2 h for immobilization. The pH and temperature optima as well as the changes in the kinetics (K_m, V_max, E_a) of the immobilized catalase was observed and discussed. Immobilized catalase preparations showed higher storage stabilities than free catalase. The half-life of free catalase, catalase immobilized via glutaraldehyde and catalase immobilized via glutaraldehyde+spacer were calculated as 2, 55 and 10 days at room temperature and 4, 85 and 107 days at 5 °C, respectively. The operational stability of the catalase immobilized via glutaraldehyde was higher than the catalase immobilized via glutaraldehyde+spacer. The remaining activity of the catalase immobilized via glutaraldehyde was about 90% and that of the catalase immobilized via glutaraldeyde+spacer was about 30% after 20 cycles of batch operation.     

Effects of Genotype on Testis, Semen Quality, and Mineral Composition of Semen in Various Ram Breeds

This study was carried out on nine 3–5-year-old rams of different species: four Norduz, two Karakas, and three Ile de France?×?Akkaraman rams. Sperm of each ram was collected by means of an artificial vagina once per month from June to October 2007. In each case, the sperm volume, pH, viscosity, mass activity, motility, concentration, abnormal spermatozoa ratio, and the sperm death/live ratio were recorder along with the animals’ testis diameter and scrotum circumferences. The concentrations of calcium, potassium, copper, magnesium, iron, and zinc levels in seminal plasma were determined. There were changes in the measured parameters according to genotype and time of sampling, which can be useful for better planning of anestrous mating programs for these ram species.     

X-linked megalocornea caused by mutations in CHRDL1 identifies an essential role for ventroptin in anterior segment development

X-linked megalocornea (MGC1) is an ocular anterior segment disorder characterized by an increased cornea diameter and deep anterior chamber evident at birth and later onset of mosaic corneal degeneration (shagreen), arcus juvenilis, and presenile cataracts. We identified copy-number variation, frameshift, missense, splice-site and nonsense mutations in the Chordin-like 1 gene (CHRDL1) on Xq23 as the cause of the condition in seven MGC1 families. CHRDL1 encodes ventroptin, a bone morphogenic protein antagonist with a proposed role in specification of topographic retinotectal projections. Electrophysiological evaluation revealed mild generalized cone system dysfunction and, in one patient, an interhemispheric asymmetry in visual evoked potentials. We show that CHRDL1 is expressed in the developing human cornea and anterior segment in addition to the retina. We explored the impact of loss of ventroptin function on brain function and morphology in vivo. CHRDL1 is differentially expressed in the human fetal brain, and there is high expression in cerebellum and neocortex. We show that MGC1 patients have a superior cognitive ability despite a striking focal loss of myelination of white matter. Our findings reveal an unexpected requirement for ventroptin during anterior segment development and the consequences of a lack of function in the retina and brain.     

Investigation of the Effects of 2.1 GHz Microwave Radiation on Mitochondrial Membrane Potential (ΔΨ m), Apoptotic Activity and Cell Viability in Human Breast Fibroblast Cells

In the present study we aimed to investigate the effects of 2.1 GHz Wideband Code Division Multiple Access (W-CDMA) modulated Microwave (MW) Radiation on cell survival and apoptotic activity of human breast fibroblast cells. The cell cultures were exposed to W-CDMA modulated MW at 2.1 GHz at a SAR level of 0.607 W/kg for 4 and 24 h. The cell viability was assessed by MTT [3-(4, 5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] method. The percentage of apoptotic cells was analyzed by Annexin V-FITC and PI staining. 5,5′,6,6′-Tetrachloro-1,1′,3,3′- tetraethylbenzimidazolcarbocyanine iodide (JC-1) was used to measure Mitochondrial Membrane Potential (ΔΨ _m). sFasL and Fas/APO-1 protein levels were determined by ELISA method. 2.1 GHz MW radiation was shown to be able to inhibit cell proliferation and induce apoptosis in human breast fibroblast cells. The cell viability of MW-exposed cells was decreased significantly. The percentages of Annexin V-FITC positive cells were higher in MW groups. ΔΨ _m was decreased significantly due to MW radiation exposure. However, neither sFas nor FasL level was significantly changed in MW-exposed fibroblast cells. The results of this study showed that 2.1 GHz W-CDMA modulated MW radiation-induced apoptotic cell death via the mitochondrial pathway.     

Plk1 controls the Nek2A-PP1γ antagonism in centrosome disjunction

In human cells, separation of the centrosomes and formation of a bipolar spindle are essential for correct chromosome segregation [1]. During interphase, centrosomes are joined together by the linker proteins C-Nap1 and rootletin [2-4]. At the onset of mitosis, these linker proteins are phosphorylated and displaced from centrosomes by the Nek2A kinase, which is regulated by two Hippo pathway components, Mst2 kinase and the scaffold protein hSav1. The kinesin-5 motor protein Eg5 promotes centrosome separation in a parallel pathway to Nek2A [5]. Here, we report that Polo-like kinase 1 (Plk1) functions upstream of the Mst2-Nek2A kinase module in centrosome disjunction as well as being important for Eg5 localization at centrosomes. Plk1 regulates Mst2-Nek2A-induced centrosome disjunction by phosphorylating Mst2. The absence of Plk1 phosphorylation of Mst2 promotes assembly of Nek2A-PP1γ-Mst2 complexes, in which PP1γ counteracts Nek2A kinase activity. In contrast, Plk1 phosphorylation of Mst2 prevents PP1γ binding to Mst2-Nek2A, allowing Nek2A activity to promote centrosome disjunction. We propose that centrosome disjunction is regulated by Plk1, providing a well-balanced control between the counteracting Nek2A and PP1γ activities on the centrosome linker.     

A unique Golgi apparatus distribution may be a marker for osteogenic differentiation of hDP-MSCs

Stem cell markers are utilized to isolate or identify stem cells. So far, many stem-cell-specific markers have been described, although some of them turned out to be not as specific as it was originally proposed. In this study, we sought to search for a specific stem cell marker that would be phenotypically helpful, characteristically specific, economically affordable and easy to use. Because organelles are one of the major characteristics of eukaryotic cells, we asked the question of whether organelle characteristics might be a useful tool for stem cell characterization. We studied distribution and characteristics of the endoplasmic reticulum, the mitochondria and the Golgi apparatus in human dental-pulp-derived mesenchymal stem cells before and during osteogenic differentiation. Although it was not possible to find a useful macromolecular marker for stem cell characterization, we found that during osteogenic differentiation, the stem cells changed their Golgi characteristics and displayed a unique in vivo pattern. We analysed these unique Golgi structures and proposed a potential osteogenic differentiation marker for human dental-pulp-derived mesenchymal stem cells. This pattern may be used in the evaluation of osteogenic differentiation.