Topoisomerase III Acts at the Replication Fork To Remove Precatenanes.

The role of DNA topoisomerase III (Topo III) in bacterial cells has proven elusive. Whereas eukaryotic Top IIIα homologs are clearly involved with homologs of the bacterial DNA helicase RecQ in unraveling double Holliday junctions, preventing crossover exchange of genetic information at unscheduled recombination intermediates, and Top IIIß homologs have been shown to be involved in regulation of various mRNAs involved in neuronal function, there is little evidence for similar reactions in bacteria. Instead, most data point to Topo III playing a role supplemental to that of topoisomerase IV in unlinking daughter chromosomes during DNA replication. In support of this model, we show that Escherichia coli Topo III associates with the replication fork in vivo (likely via interactions with the single-stranded DNA-binding protein and the ß clamp-loading DnaX complex of the DNA polymerase III holoenzyme), that the DnaX complex stimulates the ability of Topo III to unlink both catenated and precatenated DNA rings, and that ΔtopB cells show delayed and disorganized nucleoid segregation compared to that of wild-type cells. These data argue that Topo III normally assists topoisomerase IV in chromosome decatenation by removing excess positive topological linkages at or near the replication fork as they are converted into precatenanes.IMPORTANCE Topological entanglement between daughter chromosomes has to be reduced to exactly zero every time an E. coli cell divides. The enzymatic agents that accomplish this task are the topoisomerases. E. coli possesses four topoisomerases. It has been thought that topoisomerase IV is primarily responsible for unlinking the daughter chromosomes during DNA replication. We show here that topoisomerase III also plays a role in this process and is specifically localized to the replisome, the multiprotein machine that duplicates the cell's genome, in order to do so.

The MukB-topoisomerase IV interaction is required for proper chromosome compaction.

The bacterial condensin MukB and the cellular decatenating enzyme topoisomerase IV interact. This interaction stimulates intramolecular reactions catalyzed by topoisomerase IV, supercoiled DNA relaxation, and DNA knotting but not intermolecular reactions such as decatenation of linked DNAs. We have demonstrated previously that MukB condenses DNA by sequestering negative supercoils and stabilizing topologically isolated loops in the DNA. We show here that the MukB-topoisomerase IV interaction stabilizes MukB on DNA, increasing the extent of DNA condensation without increasing the amount of MukB bound to the DNA. This effect does not require the catalytic activity of topoisomerase IV. Cells carrying a mukB mutant allele that encodes a protein that does not interact with topoisomerase IV exhibit severe nucleoid decompaction leading to chromosome segregation defects. These findings suggest that the MukB-topoisomerase IV complex may provide a scaffold for DNA condensation.

3 D Network-Structured Crumpled Graphene/Carbon Nanotube/Polyaniline Composites for Supercapacitors.

Crumpled graphene (CGR) is considered a promising supercapacitor material to achieve high power and energy density because it could overcome the disadvantages of 2 D GR sheets such as aggregation during the electrode fabrication process, reduction of the available surface area, and limitation of the electron and ion transport. Even though CGR shows good results, carbon materials are limited in terms of their capacitance performance. Here, we report highly enhanced supercapacitor materials by fabricating a 3 D composite containing CGR, carbon nanotubes (CNTs), and polyaniline (PANI). The CNTs increased the basal spacing and bridged the defects for electron transfer between the GR sheets in CGR. PANI can enhance the rate of conduction of electrons and offer high pseudocapacitance originating from its redox reactions. The synergistic effect of the CNTs and PANI may also result in a higher electrochemical capacitance and better stability than each individual component as electrode materials for supercapacitors in a two-electrode system. More importantly, the performance of the supercapacitors can be further enhanced by employing 2 D GR as the binder for the composite electrodes, resulting in specific capacitance of 456 F g-1 , rate capability of 89 %, and cyclic stability of 97 % after 1000 cycles.

Effect of long-term dietary arginyl-fructose (AF) on hyperglycemia and HbA1c in diabetic db/db mice.

We have previously reported that Amadori compounds exert anti-diabetic effects by lowering sucrose-induced hyperglycemia in normal Sprague-Dawley rats. In the present study we extended our recent findings to evaluate whether α-glucosidase inhibitor arginyl-fructose (AF) lowers blood glucose level in diabetic db/db mice, a genetic model for type 2 diabetes. The db/db mice were randomly assigned to high-carbohydrate diets (66.1% corn starch) with and without AF (4% in the diet) for 6 weeks. Changes in body weight, blood glucose level, and food intake were measured daily for 42 days. Dietary supplementation of AF resulted in a significant decrease of blood glucose level (p < 0.001) and body weight (p < 0.001). The level of HbA1c, a better indicator of plasma glucose concentration over prolonged periods of time, was also significantly decreased for 6-week period (p < 0.001). Dietary treatment of acarbose® (0.04% in diet), a positive control, also significantly alleviated the level of blood glucose, HbA1c, and body weight. These results indicate that AF Maillard reaction product improves postprandial hyperglycemia by suppressing glucose absorption as well as decreasing HbA1c level.

Impact of quality parameters on the recovery of putrescine and cadaverine in fish using methanol-hydrochloric acid solvent extraction.

Methanol (MeOH) extraction by AOAC Official Method 996.07 has resulted in low amine recoveries in fresh fish tissue. Addition of 25% 0.4 M HCl to the 75% methanol-water extraction solvent resulted in higher recoveries of putrescine and cadaverine. Average putrescine recovery increased from 55 to 92% in flounder, scup, bluefish, and salmon; from 92 to 98% in mackerel; and from 83 to 107% in processed mackerel. Average cadaverine recovery increased from 57 to 95% in flounder, scup, bluefish, and salmon; from 91 to 97% in mackerel; and from 92 to 108% in processed mackerel. Fish stored on ice for 12 days also showed differences between background concentrations determined with the two solvents. However, the values decreased with storage time, indicating that degradation of the protein matrix may cause more comparable measurements between the two solvents. However, consistently higher putrescine and cadaverine measurements were determined using MeOH-HCl. Although significant differences in the extraction of amines from the high-fat fish tissue were not seen between MeOH and MeOH-HCl, it would be ideal to have one solvent for biogenic amine extraction. This study confirms that MeOH-HCl is a better solvent for complete extraction and recovery of putrescine and cadaverine in fresh and processed fish tissues.

Seasonal variation of phenolic antioxidant-mediated α-glucosidase inhibition of Ascophyllum nodosum.

Ascophyllum nodosum is a brown seaweed that grows abundantly in the US Northeast coastal region. This study examined the seasonal variation of A. nodosum in phenolic contents and subsequent antioxidant, α-glucosidase and α-amylase inhibitory activities. A. nodosum was harvested monthly and extracted in hot water and the resulting extracts were spray-dried. The results indicate a clear seasonal variation in terms of phenolic content, with June and July being the highest (36.4 and 37 mg/g, respectively) and May the lowest (21.8 mg/g). The antioxidant activities, in terms of 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging activity, correlated with the phenolic contents observed (r = 0.81), with the month of July being the highest (58%) and April the lowest (26%). Similarly in terms of Trolox equivalent, July had the highest activity (15.53 µM) and April and May the lowest (8.40 and 8.27 µM, respectively). α-glucosidase inhibitory activity exhibited a pattern similar to the phenolic contents observed with July having the highest inhibitory activity (IC(70) 2.23 µg) and April the lowest (IC(70) 26.13 µg), resulting in an inverse correlation between IC(70) values and total phenolic content (r = -0.89). Such seasonal variation is believed to be caused by temperature-related stress considering that A. nodosum is a cold water species.

Physical and oxidative stabilization of omega-3 fatty acids in surimi gels.

UNLABELLED: The objective of the study was to compare the dispersion and oxidative stability of omega-3 fatty acid oil in high- and low-quality surimi gels during 4-mo refrigerated and frozen storage. Low-quality surimi was prepared by subjecting Alaska pollock surimi to 7 freeze-thaw cycles. Surimi gels were prepared with 4% modified starch, 2% salt, and 0.5% or 1% algal DHA or concentrated fish EPA-DHA oil, and stored at -18 or 3 degrees C for 4 mo after being vacuumed packed and pasteurized. The effect of surimi gel properties on oil dispersion was examined using light microscopy equipped with image process software. The extent of lipid oxidation was monitored by thiobarbituric acid reactive substances (TBARS), peroxide value (PV), and fatty acid methly esters (DHA and EPA). Very fine and uniform oil dispersion was observed in the high-quality surimi gel with the average droplet size of 12.37 microm(2) and dispersion of 1.73 x 10(-3) droplets/microm(2) compared to 84.32 microm(2) and 0.57 x 10(-3) droplets/microm(2) in the low-quality gel. Throughout the 4 mo storage, TBARS and PV of high-quality surimi gel were significantly (P < 0.05) lower than those of low-quality surimi gel. The decreases in omega-3 fatty acids in the high-quality surimi gels were lower than those in the low-quality surimi gels under both storage conditions. Results confirm that a highly cohesive gel matrix is required to have a fine dispersion and oxidative stability of omega-3 fatty acids in the surimi gel system. PRACTICAL APPLICATION: Uniform dispersion and oxidative stability of omega-3 fatty acid oil can be achieved in the highly cohesive surimi gel system without use of antioxidants. This suggests that surimi can be used as a protein-based carrier in developing high omega-3 fatty acids-containing seafood products.

Effect of the molecular mass and degree of substitution of oleoylchitosan on the structure, rheological properties, and formation of nanoparticles.

Oleoylchitosans (O-chitosans), with different molecular masses and degrees of substitution (DS), were synthesized by reacting chitosan with oleoyl chloride. The FT-IR suggested the formation of an amide linkage between amino groups of chitosan and carboxyl groups of oleic acid. The viscosity of O-chitosan sharply increased with the increase of concentration, whereas that of unmodified chitosan rose only slightly. This increase was stronger as the increase of hydrophobicity (DS) and molecular mass of the polymer. The critical aggregation concentration (CAC) of O-chitosans with DS 5, 11, and 27% were 79.43, 31.6, 10 mg/L, respectively, and the CAC of samples with molecular masses of 20, 38, 300, and 1100 kDa were 50.1, 74.93, 125.9, and 630.9 mg/L, respectively. All of the O-chitosans could reduce surface tension slightly. Nanoparticles were prepared using an O/W emulsification method. Mean diameters of the polymeric amphiphilic nanoparticles of O-chitosans with DS 5 and 11% were around 327.4 and 275.3 nm, respectively.

O/W emulsification for the self-aggregation and nanoparticle formation of linoleic acid-modified chitosan in the aqueous system.

Chitosan was modified by coupling with linoleic acid through the 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide-mediated reaction to increase its amphipathicity for improved emulsification. The micelle formation of linoleic acid-modified chitosan in the 0.1 M acetic acid solution was enhanced by O/W emulsification with methylene chloride, an oil phase. The fluorescence spectra indicate that without emulsification the self-aggregation of LA-chitosan occurred at the concentration of 1.0 g/L or above, and with emulsification, self-aggregation was greatly enhanced followed by a stable micelle formation at 2.0 g/L. The addition of 1 M sodium chloride promoted the self-aggregation of LA-chitosan molecules both with and without emulsification. The micelles of LA-chitosan formed nanosize particles ranging from 200 to 600 nm. The LA-chitosan nanoparticles encapsulated the lipid soluble model compound, retinal acetate, with 50% efficiency.

Effect of gamma-irradiation on degradation of alginate.

The aqueous solution of alginate was irradiated by 60Co gamma-rays in the dose range of 10-500 kGy. To assess the effect of irradiation on the degradation of alginate, the irradiation-induced changes in the viscosity, molecular weight, color, monomer composition, and sequence were measured. The molecular weight of raw alginate was reduced from 300000 to 25000 when irradiated at 100 kGy. The degradation rate decreased and the chain breaks per molecule increased with increasing irradiation dose. The viscosity of irradiated alginate solution reached a near minimum as low as at 10 kGy. No appreciable color changes were observed in the samples irradiated at up to 100 kGy, but intense browning occurred beyond 200 kGy. The 13C NMR spectra showed that homopolymeric blocks, MM and GG, increased and the M/G ratio decreased with irradiation. Considering both the level of degradation and the color change of alginate, the optimum irradiation dose was found to be 100 kGy.