Characterization and properties of catalase immobilized onto controlled pore glass and its application in batch and plug-flow type reactors

Bovine liver catalase was covalently immobilized onto controlled pore glass (CPG) beads modified with 3-aminopropyltriethoxysilane (3-APTES) followed by treatment with glutaraldehyde. Coupling of catalase onto CPG was optimized to improve the efficiency of the overall immobilization procedure. The optimum coupling conditions: pore diameter of CPG, pH, buffer concentration, temperature, coupling time and initial catalase amount per grams of carrier were determined as 70 nm, 6.0, 75 mM, 5 °C, 7 h and 6 mg catalase, respectively. Catalytic efficiencies (k_cat/K_m) and thermal inactivation rate constants (k_i) of ICPG1 were determined and compared with that of free catalase. Suitability of ICPG1 was also investigated by using it in batch and plug-flow type reactors. When the remaining activity of ICPG1 retained was about 50% of its initial activity the highest total productivity of ICPG1 was determined as 7.6 × 10⁶ U g immobilized catalase⁻¹ in plug-flow type reactor. However, the highest total productivity of ICPG1 was 6.2 × 10⁵ U g immobilized catalase⁻¹ in batch type reactor. ICPG1 may have great potentials as biocatalyst for the application in decomposition of hydrogen peroxide in plug-flow type reactor.     

The role of the ADRA2A C1291G genetic polymorphism in response to dexmedetomidine on patients undergoing coronary artery surgery

The existence of interindividual drug response variability has been known for a long time. Individual susceptibility which might cause toxicity or inadequate treatment is important in drug therapy. Genetic polymorphisms in genes responsible for drug response are expected to be useful in keeping track of differences among individuals. Dexmedetomidine is a sedative drug, whose use in intensive care unit patients was confirmed by USA-Food Drug Administration (FDA) by the end of 1999. It was proven that dexmedetomidine shows its clinic effect via the α₂-AR. However, to the best of our knowledge, to date, there is no investigation in clinic indicating the relation between dexmedetomidine and α_2A-AR gene polymorphism. The aim of our study was to investigate the association between the effect of α_2A-Adrenergic Receptor (ADRA2A) C-1291G gene polymorphism in the promoter region of the candidate gene and clinical effects (sedative and haemodynamics effects) of dexmedetomidine. One hundred and ten patients undergoing coronary artery surgery were prospectively studied. Anesthetic technique was standardized with fentanyl, midazolam and rocuronium bromide. Patients were monitorized with Bispectral Index (BIS) monitor in addition to the routine invasive haemodynamic monitorization in the operation room. The Ramsay Sedation Scale was also used in order to determine the sedation level just arriving to Intensive Care Unit (ICU). The genotyping of ADRA2A C1291G was done by Restriction Fragment Length Polymorphism-Polymerase Chain Reaction (RFLP-PCR). We found the frequencies of C1291C, C1291G and G1291G genotypes, as 43.6, 45.5 and 10.9%, respectively. Patients who carry variant genotype had higher BIS and Ramsay Sedation Scores, indicating a longer period for falling asleep. The results of our study are promising, considering the association between ADRA2A polymorphism and response to dexmedetomidine. However, further investigations on other ADRA2A locus or haplotypes might be useful to clarify the relation between this gene and dexmedetomidine activity.     

Separate to operate: control of centrosome positioning and separation

The centrosome is the main microtubule (MT)-organizing centre of animal cells. It consists of two centrioles and a multi-layered proteinaceous structure that surrounds the centrioles, the so-called pericentriolar material. Centrosomes promote de novo assembly of MTs and thus play important roles in Golgi organization, cell polarity, cell motility and the organization of the mitotic spindle. To execute these functions, centrosomes have to adopt particular cellular positions. Actin and MT networks and the association of the centrosomes to the nuclear envelope define the correct positioning of the centrosomes. Another important feature of centrosomes is the centrosomal linker that connects the two centrosomes. The centrosome linker assembles in late mitosis/G1 simultaneously with centriole disengagement and is dissolved before or at the beginning of mitosis. Linker dissolution is important for mitotic spindle formation, and its cell cycle timing has profound influences on the execution of mitosis and proficiency of chromosome segregation. In this review, we will focus on the mechanisms of centrosome positioning and separation, and describe their functions and mechanisms in the light of recent findings.     

Effects of Lithium and Lamotrigine on Oxidative–Nitrosative Stress and Spatial Learning Deficit After Global Cerebral Ischemia

Lithium (Li) and lamotrigine (LTG) have neuroprotective properties. However, the exact therapeutic mechanisms of these drugs have not been well understood. We investigated the antioxidant properties of Li (40 and 80 mg/kg/day) and LTG (20 and 40 mg/kg/day) in a rat model of global cerebral ischemia based on permanent bilateral occlusion of the common carotid arteries (BCAO). Nitric oxide (NO), malondialdehyde (MDA), glutathione (GSH), glutathione reductase (GSH-R), catalase (CAT) and superoxide dismutase (SOD) levels were measured as an indicator of oxidative–nitrosative stress in both prefrontal cortex (PFC) and hippocampus after 28 days of treatment. The spatial learning disability was also assessed at the end of the study by Morris water maze (MWM) test. All oxidative–nitrosative parameters were found to be higher in the groups under treatment than in sham. Both drugs caused a decrease in PFC NO and MDA elevation, meanwhile the increase in GSH, GSH-R, CAT and SOD levels was significantly more evident in treated groups. We also found higher PFC GSH-R and hippocampal SOD levels in BCAO + Li (80 mg/day) treated group when compared with BCAO + LTG 40 mg/day. MWM test data showed a similar increase in spatial learning ability in all groups under treatment. We found no other statistical difference in comparison of treated groups with different dosages. Our findings suggested that Li and LTG treatments may decrease spatial learning memory deficits accompanied by lower oxidative–nitrosative stress in global cerebral ischemia. Both drugs may have potential benefits for the treatment of vascular dementia in clinical practice.     

Remote sensing as a tool for assessing water quality in Loosdrecht lakes

The underwater light field in 7 lakes in the Loosdrecht lake area was measured in situ. Subsurface upwelling irradiance and irradiance reflectance, together with estimations of scattering and laboratory measurements of absorption by aquatic humus and particulate matter, enabled an analysis of the spectral signature of these waters. Aircraft imaging spectrometer measurements of upwelling radiance at 1 km altitude were used to simulate the PMI Chlorophyll #1, the CAESAR Inland Water Mode spectral bandsets and the Thematic Mapper bands 1 to 4. This made it possible to compare the effects of spectral band width and selection on the estimation of water quality parameters. Correlations increased to r > 0.94, at a significance level of 1% for the simulated C-IWM data with the 6 water quality parameters. Images of the PMI Chlorophyll #1 and of the TM were analysed and found to be in accordance with the statistical modelling results.A significant increase in correlation of remote sensing data with water quality parameters can be achieved through the selective use of 10 to 20 nm wide bands in the spectral range of 500 to 720 nm in these eutrophic waters. Sum of chlorophyll a and phaeopigments, seston dry weight, Secchi disc transparency, and coefficients for vertical attenuation of light, absorption and scattering can be estimated accurately. TM image data for water quality assessment is of limited use due to the relatively low spectral and radiometric resolution. However, the revisit capability and relatively low price per area are positive aspects of these satellite images.Abbreviations CAESAR = CCD Airborne Experimental Scanner for Applications in Remote sensing - C-IWM = CAESAR Inland Water Mode - CCD = charge coupled device - EOS-A = Earth Observation System Platform A - PAR = photosynthetically active radiation from 400–700 nm. - PMI = Programmable Multispectral Imager - RSLL = Remote Sensing Loosdrecht Lakes Project - SPOT = Systeme Pour l'Observation de la Terre - SPOT-HRV = Sensor on board of the SPOT satellite - TM = Thematic Mapper instrument aboard the Landsat 5 satellite     

Unilateral gate flap for reconstruction of the lower lip

A unilateral gate-flap technique consisting of a nasolabial island flap is presented for the reconstruction of defects in the lower lip after excision of large, laterally located epidermoid tumors. The amount of healthy tissue resected is optimal. The reconstructed lower lip retains sensation and muscle function and is continent with a satisfying appearance. Temporary flap edema and a vermilion notch at the apex of the flap are both avoidable problems. This method may be used in selected patients with large advanced epidermoid cancers of the lower lip.     

Promising anticancer activity of a lichen,Parmelia sulcata Taylor,against breast cancer cell lines and genotoxic effect on human lymphocytes

Plants are still to be explored for new anti-cancer compounds because overall success in cancer treatment is still not satisfactory. As a new possible source for such compounds, the lichens are recently taking a great attention. We, therefore, explored both the genotoxic and anti-growth properties of lichen species Parmelia sulcata Taylor. The chemical composition of P. sulcata was analyzed with comprehensive gas chromatography–time of flight mass spectrometry. Anti-growth effect was tested in human breast cancer cell lines (MCF-7 and MDA-MB-231) by the MTT and ATP viability assays, while the genotoxic activity was studied by assays for micronucleus, chromosomal aberration and DNA fragmentation in human lymphocytes culture. Cell death modes (apoptosis/necrosis) were morphologically assessed. P. sulcata inhibited the growth in a dose-dependent manner up to a dose of 100 μg/ml and induced caspase-independent apoptosis. It also showed genotoxic activity at doses (>125 μg/ml) higher than that required for apoptosis. These results suggest that P. sulcata may induce caspase-independent apoptotic cell death at lower doses, while it may be genotoxic at relatively higher doses.     

Characterization of Exopolysaccharides (EPSs) Obtained from Ligilactobacillus salivarius Strains and Investigation at the Prebiotic Potential as an Alternative to Plant Prebiotics at Poultry

In this study, it was aimed to reveal the potential of using exopolysaccharides (EPS) obtained from Ligilactobacillus salivarius as a prebiotic that regulates chicken intestinal microbiota. Characterization of EPS obtained from L. salivarius BIS312 (EPSBIS312) and BIS722 (EPSBIS722) strains was demonstrated by high performance liquid chromatography (HPLC), nuclear magnetic resonance (NMR), and size-exclusion chromatography (SEC) analyses. It was determined that the molecular weight of both EPS is in the range of 10⁴–10⁶ Daltons, and there are 4 types of monomers in their structure. Anti-biofilm and anti-quorum sensing effects of EPSBIS312 and EPSBIS722 were determined. EPSBIS312 and EPSBIS722 showed a strong anti-biofilm effect on Enterococcus faecalis ATCC 29212, Staphylococcus aureus EB-1, and Escherichia coli ATCC 11229. The anti-quorum sensing study revealed that the EPSBIS722 had a higher effect than the EPSBIS312. The effect of different concentrations of EPS (2.5%, 5%, 10%) on lactobacilli growth stimulator (LGS) was evaluated. The highest LGS was promoted at 10% concentration while the lowest LGS was promoted at 2.5% concentration by EPSBIS722. In addition, adhesion abilities of EPSBIS312 and EPSBIS722 in HT-29 colorectal adenocarcinoma cell line were tested. EPSs significantly increased the ability to adhere to HT-29 cells. The characterized EPSs may be an alternative to plant prebiotics such as inulin at poultry.

     

RNA interference-mediated inhibition of Semliki Forest virus replication in mammalian cells

RNA interference (RNAi) has recently shown promise as a mode of inhibition of slowly replicating viruses causing chronic diseases such as hepatitis C. To investigate whether RNAi is also feasible for rapidly growing RNA viruses such as alphaviruses, we tested the ability of expressed short hairpin RNAs (shRNAs) to inhibit the Semliki Forest virus (SFV), a rapidly replicating positive-strand RNA virus. Plasmids expressing shRNAs targeting SFV target sequences under the control of a human U6 promoter were introduced into BHK-21 cells. The targets included sequences encoding nonstructural (nsP1, 2, and 4) and structural (capsid) proteins as well as nonviral sequences serving as control targets. Twenty-four to 48 hours following transfection with shRNA plasmids, the cells were infected with replication-competent or replication-deficient recombinant SFV expressing green fluorescent protein (GFP) at a multiplicity of infection (MOI) of approximately 5. Viral replication was monitored by fluorescence microscopy and flow cytometry. Specific and marked reduction of viral replication was observed with shRNAs targeting nsP1 and nsP4. The degree of inhibition of the replication-deficient SFV was >or=70% over a 5-day period, a level similar to the transfection efficiency, suggesting complete inhibition of nonreplicating virus in the transfected cell population. However, only nsP1 shRNA was inhibitory against replication-competent SFV (approximately 30%-50% reduction), and this effect was transient. No inhibition was observed with control shRNAs. In contrast to the recent success of RNAi approaches for slowly growing viruses, these results illustrate the challenge of inhibiting very rapidly replicating RNA viruses by RNAi. However, the addition of RNAi approaches to other antiviral modalities might improve the response to acute infections.