[Research on Hemorheology in Rats with Acute Hyperuricemia].

Hyperuricemia is a risk factor for various diseases, but knowledge on acute hyperuricemia is still not sufficient. The present study was aimed to investigate the effect of acute hyperuricemia on red blood cells from hemorheological point of view, and to provide the reference for clinical treatment. The rats were gavaged with 500 mg/kg hypoxanthine and intraperitoneally injected with 100 mg/kg oxonate to induce the model of acute hyperuricemia. The same volume of blood samples were drawn within time period of 0, 1, 2, 3 and 6 h, respectively, from the inner canthus of rats to measure the serum uric acid, hemorheological parameters and the malondialdehyde level. It was found that in each period of 1, 2 and 3 h, the rats had significantly higher levels of uric acid. The integrated deformation index and relax index were increased. The hemolysis rate was significantly reduced. The plasma malondialdehyde level was obviously decreased at the end of 2 h. The results suggested that short-term elevated uric acid could improve the hemorheological parameters and the lipid oxidative level in red blood cells.

UVC mutagenicity is suppressed in Japanese miso-treated human RSa cells, possibly via GRP78 expression.

Little is known about the ability of miso, to modulate mutability in human cells. We have observed increased levels of glucose-regulated protein 78 (GRP78) expression in association with suppression of mutation in human RSa cells irradiated with ultraviolet C (UVC). Here we examined to determine whether miso treatment results in increased GRP78 expression and suppression of UVC mutagenicity in RSa cells. Supernatants of water extracts of miso products and their components were tested. In the sample-treated cells, the amount of GRP78, as estimated by RT-PCR and immunoblotting analysis, increased, and the UVC-induced ouabain resistant mutation (Oua(R)) and the K-ras codon 12-base substitution mutation frequency decreased. This decrease was not observed in cells with downregulation of GRP78 by GRP78 siRNA transfection. The results suggest that miso suppresses UVC mutagenicity by increasing GRP78 expression in human cells.

The complete mitochondrial genome of soft coral Sinularia penghuensis Ofwegen and Benayahu, 2012 (Octocorallia: Alcyonacea): the analysis of mitogenome organization and phylogeny.

The complete mitochondrial genome of Sinularia penghuensis was sequenced and analyzed using next-generation sequencing. The present mitochondrial genome was 18730 bp in length, containing 14 protein-coding genes (PCGs) (cox1-cox3.nad1-nad6, nad4L, atp6, atp8, cytb, and MutS), two ribosomal RNA genes (rRNAs) (12S and 16S), and one transfer RNA gene (Met-tRNA). The phylogenetic analysis of family Alcyoniidae revealed that S. penghuensis and Sinularia maxima cluster together. Five species in Sinularia reveals high identity in mitogenome sequences that the lowest variable sites (SNPs) were found between S. penghuensis and S. maxima.

Lifeceramics-treated water reduces serum uric acid levels and improves hemorheological activity in hyperuricemic rats.

Lifeceramics (LC) is made of zeolite and oyster shell and is hypothesized to act as an anti-oxidative agent. In the present study, the effects of LC-treated water (LC water) on the concentration of serum uric acid (SUA) and the hemorheological parameters in male rats with hyperuricemia (HUA) was assessed. To prepare LC water, distilled water was mixed with LC particles. HUA was induced in rats by daily potassium oxonate (PO) injection (250 mg/kg). The PO-injected rats were separated into three different groups and were administered distilled water (PO rats), allopurinol [a xanthine oxidase (XOD) inhibitor] solution [PO + allopurinol (AP) rats] or LC water (PO+LC rats) by gavage. Control rats were intraperitoneally injected with sodium carboxymethyl cellulose solution and administered untreated distilled water by gavage. After injection and gavage for 5 weeks, the SUA concentration, hemorheology index and antioxidant index were measured. The SUA concentration and blood deformation index of the PO rats were significantly higher and lower, respectively, compared with the control rats. However, in the PO+LC rats, the SUA concentration and blood deformation index decreased and increased, respectively, to a level similar to that of the control as well as that in the PO+AP rats. Furthermore, the PO-induced increase in XOD activity was suppressed by combined treatment with LC water, resulting in a decrease in malondialdehyde concentration. These results suggest that LC water can reduce the SUA concentration, increase serum antioxidant activity and improve hemorheological activity in hyperuricemic rats.

The roles of HSP27 and annexin II in resistance to UVC-induced cell death: comparative studies of the human UVC-sensitive and -resistant cell lines RSa and APr-1.

We have reported that heat shock protein 27 (HSP27) and annexin II are involved in the protection of human cells against UVC-induced cell death. In this study we tried to confirm the combined roles of HSP27 and annexin II in cell death after UVC irradiation. In RSa cells with sensitivity to UVC, expression of annexin II decreased after UVC irradiation, but not in AP(r)-1 cells with increased resistance to UVC. HSP27 siRNA-transfected AP(r)-1 cells were sensitized to UVC lethality and showed decreased annexin II expression after UVC irradiation. In contrast, transfection of RSa cells with HSP27 cDNA increased their resistance to UVC lethality and caused increased annexin II expression. Furthermore, over-production of annexin II in RSa cells resulted in increased resistance to UVC lethality. This study indicates the involvement of cellular HSP27 expression in the UVC susceptibility of human cells, which occurs in association with regulation of annexin II expression.

Synthesis and characterization of chromium complexes 2-Me4CpC6H4CH2(R)NHCrCl2 and their catalytic properties in ethylene homo- and co-polymerization.

A series of new half-sandwich secondary amine-coordinated dichlorochromium complexes chelated by 2-(tetramethylcyclopentadienyl)benzylamine ligands, 2-Me4CpC6H4CH2(R)NHCrCl2 [R = iPr (1), Cy (2), Ph (3), 4-MePh (4), 2,6-Me2Ph (5), 2,6-Et2Ph (6)], have been synthesized from the reactions of CrCl3(THF)3 with the dilithium salts of the corresponding ligands in THF, followed by the addition of 1/2 eq. of H2O to the reaction mixtures. The isolated yields of the chromium complexes were found to increase with the increase in the amount of H2O introduced and reach the highest values (66-76%) when 1/2 eq. of H2O is added. Attempts to isolate the 2-(tetramethylcyclopentadienyl)benzylamidochromium complexes, 2-Me4CpC6H4CH2(R)NCrCl, were not successful. The new dichlorochromium complexes were characterized by IR, 1H NMR, EPR, and UV/Vis spectroscopy and elemental analyses, and the molecular structures of complexes 1, 5 and 6 were determined by X-ray crystallography. The X-ray crystallographic analysis reveals that these chromium complexes possess a three-legged piano-stool geometry with the amine N atom in a mitered six-membered chelating ring and the two chloride atoms as the legs. Upon activation with AlR3 and Ph3CB(C6F5)4, complexes 1-6 exhibit reasonable catalytic activity for ethylene polymerization and copolymerization with 1-hexene, producing polyethylenes with moderate to high molecular weights and poly(ethylene-co-1-hexene)s with moderate comonomer incorporation which are typical linear low-density polyethylenes (LLDPE). Complex 4 was found to show higher catalytic activity for ethylene homo- and co-polymerization than other complexes under similar conditions, while complex 3 produced poly(ethylene-co-1-hexene)s with the highest comonomer incorporation.

Half-sandwich rare-earth metal complexes bearing a C5Me4-C6H4-o-CH2NMe2 ligand: synthesis, characterization and catalytic properties for isoprene, 1-hexene and MMA polymerization.

A new ortho-dimethylaminomethylphenyl-tetramethylcyclopentadienyl ligand C5Me4H-C6H4-o-CH2NMe2 (HL) and a series of rare-earth metal complexes bearing this ligand were synthesized. Of these complexes, two binuclear alkyl complexes [(C5Me4-C6H4-o-CH2N(Me)CH2-µ)Ln(CH2SiMe3)]2 (Ln = Sc (1a) and Y (1b)) were obtained from the alkane elimination reaction of the free ligand with Ln(CH2SiMe3)3(THF)2, followed by an intramolecular C-H activation process of a NMe group in the ligand with a CH2SiMe3 group, two binuclear dichloro complexes (C5Me4-C6H4-o-CH2NMe2)2Y2Cl4[LiCl(THF)2] (2a) and [(C5Me4-C6H4-o-CH2NMe2)LuCl(µ-Cl)]2 (2b) were synthesized by the reaction of anhydrous yttrium or lutetium trichloride with the lithium salt of the ligand LiL, and the binuclear bis(borohydrido) complexes [(C5Me4-C6H4-o-CH2NMe2)Ln(µ-BH4)BH4]2 (Ln = Sm (3a) and Nd (3b)) were synthesized by the reaction of Ln(BH4)3(THF)3 (Ln = Sm and Nd) with the lithium salt of the ligand. The molecular structures of all complexes 1a, 1b, 2a, 2b, 3a and 3b were determined by single-crystal X-ray crystallography. Upon activation with AlR3/Ph3CB(C6F5)4, MAO or MMAO, the binuclear alkyl complexes 1a and 1b show good catalytic activity for isoprene cis-1,4 enriched regioselective polymerization and moderate catalytic activity for 1-hexene polymerization. Complexes 3a and 3b were studied as catalysts for methyl methacrylate polymerization reaction under different conditions and were found to show moderate to high catalytic activity.

The complete mitochondrial genome of soft coral Sarcophyton trocheliophorum (Cnidaria: Anthozoa) using next-generation sequencing.

The complete mitochondrial genome of Sarcophyton trocheliophorum was completed using next-generation sequencing (NGS) method. The mitochondrial genome is a circular molecule of 18,508 bp in length, containing 14 protein-coding genes, two ribosomal RNA genes and one transfer RNA gene (Met-tRNA). The base composition is 30.45% A, 16.03% C, 19.13% G, and 34.40% T, with an A + T content of 64.85%. A phylogenetic analysis of Alcyoniidae showed that genus Sarcophyton had the closest relationship with Sinularia.

Titanium and zirconium complexes bearing new tridentate [OSO] bisphenolato-based ligands: synthesis, characterization and catalytic properties for alkene polymerization.

A number of new sulfur-bridged tridentate [OSO] bisphenolato-based ligand precursors S(2-CH2-4-tBu-6-R-C6H2OH)2 [R = CMe3 (H2L1), CMe2Ph (H2L2), CMePh2 (H2L3), CPh3 (H2L4), and C(p-Tol)3 (H2L5)] were synthesized by reactions of Na2S·9H2O with 2 eq. of the corresponding 2-(bromomethyl)-4-(tert-butyl)-6-R-phenol. Their neutral titanium complexes [S(2-CH2-4-tBu-6-R-C6H2O)2]TiCl2 [R = CMe3 (1), CMe2Ph (2), CMePh2 (3), CPh3 (4), and C(p-Tol)3 (5)] were synthesized in high yields by direct HCl-elimination reactions of TiCl4 with the corresponding ligand precursors in toluene. Ionic titanium complexes [NHEt3][S(2-CH2-4-tBu-6-R-C6H2O)2TiCl3] [R = CMe3 (6), CMePh2 (7)] and [NH2Et2][S(2-CH2-4-tBu-6-R-C6H2O)2TiCl3] [R = CMe3 (8) and CMePh2 (9)] were obtained in high yields from the reactions of TiCl4 with the corresponding ligand precursors in the presence of 2 eq. of triethylamine or diethylamine. Neutral zirconium complexes [S(2-CH2-4-tBu-6-R-C6H2O)2]ZrCl2(THF) [R = CMe2Ph (10·THF), and CMePh2 (11·THF)] were synthesized by reactions of ZrCl4 with 1 eq. of the dilithium salt of the corresponding ligand precursors Li2L in THF. The new titanium and zirconium complexes were characterized by 1H and 13C NMR, IR and elemental analyses. The molecular structures of complexes 4, 6 and 10·THF were determined by single-crystal X-ray diffraction analysis. The X-ray crystallography analysis reveals that titanium complex 4 has a five-coordinating environment surrounding the central metal atom, while the titanium complex 6 and the THF-solvated zirconium complex 10·THF possess a six-coordinating pseudo-octahedral environment around the central metal atom. Upon activation with MAO or AliBu3/Ph3CB(C6F5)4, all these titanium and zirconium complexes exhibit moderate to high catalytic activities for ethylene polymerization and ethylene/1-hexene copolymerization with moderate to high comonomer incorporation, and the ionic titanium complexes 6, 7, 8 and 9 show lower catalytic activity than their corresponding neutral complexes under similar conditions.

Annexin II, a novel HSP27-interacted protein, is involved in resistance to UVC-induced cell death in human APr-1 cells.

Heat shock protein 27 (HSP27) is implicated in diverse biologic functions as a molecular chaperone. We found that HSP27 is involved in the protection of human cells against UVC lethality. To elucidate the molecular mechanisms underlying UVC resistance, we searched for HSP27-interacted proteins related to resistance in UVC-resistant human cells, APr-1. Three candidates for HSP27-interacted proteins were found from cell lysates using an affinity column coupled with GST-fused HSP27 protein. Interaction between HSP27 and two candidates, annexin II and HSP70, was confirmed by immunoprecipitation analysis. After UVC irradiation, the amount of the complex of HSP27 and annexin II decreased in the postnuclear fraction, while it increased in the nuclear fraction. Cells transfected with annexin II-siRNA were more susceptible to UVC lethality. These results suggest that annexin II is a novel HSP27-interacted protein which is involved in UVC resistance in human cells, at least those tested here.

Involvement of heat shock protein 27 in the susceptibility of KT human breast cancer cells to UVC and interferon lethality.

Revealing the key molecules regulating the stress-response pathways in human cells is an intriguing problem. Chaperones, such as glucose-regulated protein 78 (GRP78) and heat shock protein 27 (HSP27), are important molecules for protecting the viability of human cells; however, it remains to be further clarified whether the molecules differentially modulate cellular responses to various types of stressors, such as DNA-damaging ultraviolet ray C (principally 254-nm wavelength, UVC) and cytocidal cytokine interferons. In the present study, the human breast cancer cell lines KT and MCF-7 were examined for GRP78 and HSP27 expression following exposure to UVC and human interferon-ß (HuIFN-ß). The KT cells demonstrated a higher sensitivity to both UVC and HuIFN-ß lethality than MCF-7 cells. The cellular expression levels of GRP78 in KT cells, assessed by western blot analysis, were approximately 2-fold higher than that in MCF-7 cells, while the expression of HSP27 in the KT cells was 20% of the expression in the MCF-7 cells. Decreased resistance to UVC lethality was observed in GRP78 siRNA-transfected KT cells. In addition, HSP27 cDNA transfection of KT cells resulted in an increased resistance to UVC lethality. The cDNA-transfected KT cells showed an increased viability against HuIFN-ß, compared with that of empty vector-transfected cells. By contrast, KT cells pretreated with HuIFN-ß and irradiated with UVC demonstrated an increased resistance to UVC lethality, in association with increased levels of HSP27 expression. Thus, HSP27 may control the survival response pathways to both UVC and HuIFN-ß in the human cells examined.

Extracellular recombinant annexin II confers UVC-radiation resistance and increases the Bcl-xL to Bax protein ratios in human UVC-radiation-sensitive cells.

In this study, we found that refractoriness to ultraviolet (UVC) light-induced cell death was increased in UVC-radiation-sensitive cells derived from Cockayne syndrome patients when the cells were precultured in medium supplemented with recombinant annexin II (rANX II). In CS3BES cells, an immortal cell line derived from Cockayne syndrome patients, the rANX II supplementation-induced UVC-radiation resistance was suppressed by treatment with an anti-annexin II antibody and EGTA. The amount of biotinylated annexin II on the cell surface increased in the rANX II-supplemented cells but did not increase in the cells that were cotreated with rANX II and EGTA. The capacity to remove UVC-radiation-damaged DNA, (6-4) photoproducts and cyclobutane pyrimidine dimers, was the same in cells that were precultured with rANX II and in control cells that did not receive rANX II supplementation. The rANX II supplementation-induced UVC-radiation resistance was also observed in nucleotide excision repair-deficient cells and xeroderma pigmentosum group A-downregulated cells. The Bcl-xL to Bax protein ratios, an index of survival activity in cells exposed to lethal stresses, were increased in the cells that had been precultured in rANX II for 24 h prior to UVC irradiation. Treatment with a phosphatidylinositol 3-kinase inhibitor suppressed the increased UVC-radiation resistance and Bcl-xL to Bax ratios in the cells with rANX II supplementation. Furthermore, downregulation of Bcl-xL by siRNA transfection also suppressed the UVC-radiation resistance that was induced by rANX II supplementation. These results suggest that the increase in the Bcl-xL to Bax ratios may be associated with enhanced resistance to UVC-radiation-induced cell death.

Leupeptin-sensitive proteases involved in cell survival after X-ray irradiation in human RSa cells.

Proteases have received attention as important cellular components responsible for stress response in human cells. However, little is known about the role of proteases in the early steps of cell response after X-ray irradiation. In the present study, we first searched for proteases whose activity levels are changed soon after X-ray irradiation in human RSa cells with a high sensitivity to X-ray cell-killing. RSa cells showed an increased level of fibrinolytic protease activity within 10 min after irradiation with X-ray (up to 3 Gy). The induced protease activity was proved to be inhibited by leupeptin. We next examined whether this protease inducibility is related to the X-ray susceptibility of cells. Treatment of RSa cells with leupeptin prior to X-ray irradiation resulted in lowered colony survival and an increased ratio of G(2)/M-arrested cells and apoptotic cells. These results suggest that leupeptin-sensitive proteases are involved in the resistance of human RSa cells to X-ray cell-killing.