Cloning and Expression of NS3 CDNA Fragment of HCV Genome of Hebei Isolate in E.coli

World Journal of Gastroenterology : WJG. Apr, 1998  |  Pubmed ID: 11819265

AIM:To obtain greater antigenicity of HCV NS3 protein.METHODS:The HCV NS3 cDNA fragment was amplified by reverse transcription polymerase chain reaction from the sera of the HCV infected patients.The DNA sequence was determined by dideoxy-mediated chain termination method using T7 polymerase.HCV NS3 protein was expressed in E. coli.RESULTS:Sequence analysis indicated that the HCV isolate of this study belongs to HCV-II; SDS-PAGE demonstrated an M(r) 23800 and an M(r) 22000 recombinant protein band which amount to 14% and 11% of the total bacterial proteins separately.Western blotting and ELISA showed NS3 protein possessed greater antigenicity.CONCLUSION:Recombinant HCV NS3 protein was expressed successfully, which provided the basis for developing HCV diagnostic reagents.

Evidence for Expression of Melanocortin-1 Receptor in Human Sebocytes in Vitro and in Situ

The Journal of Investigative Dermatology. Mar, 2002  |  Pubmed ID: 11874495

Many lines of evidence indicate that the activity of sebaceous glands can be modulated by neuropeptides. Direct evidence in man, however, is still missing. We show that SZ95 sebocytes, an immortalized human sebaceous gland cell line, express receptors for alpha-melanocyte-stimulating hormone. Reverse transcription polymerase chain reaction with primers against the five melanocortin receptors and immunofluorescence studies using an antibody directed against a peptide corresponding to the amino acids 2-18 of the human melanocortin-1 receptor disclosed specific transcripts and immunoreactivity for melanocortin-1 receptor in these cells. Melanocortin-1 receptor expression was confirmed in sebocytes of normal human skin by immunohistochemistry. In contrast, no immunostaining for the melanocortin-5 receptor could be detected in sebocytes in situ, in accordance with the lack of specific transcripts for this melanocortin receptor in SZ95 sebocytes. As cytokines play an important role in the recruitment of inflammatory cells in acne and related disorders and alpha-melanocyte-stimulating hormone exerts immunomodulatory effects in many other cell types, we investigated the effect of alpha-melanocyte-stimulating hormone on interleukin-8 secretion by SZ95 sebocytes. Treatment with interleukin-1beta resulted in a marked increase in interleukin-8 release that was partially blocked by coincubation with alpha-melanocyte-stimulating hormone in a dose-dependent manner. Taken together, we show here that the melanocortin-1 receptor is expressed in vitro and in situ in human sebocytes. By modulating interleukin-8 secretion, alpha-melanocyte-stimulating hormone may act as a modulator of inflammatory responses in the pilosebaceous unit.

[Analysis of Fetal Distress in Pregnancy with Hepatitis B Virus Infection]

Zhonghua Fu Chan Ke Za Zhi. Apr, 2002  |  Pubmed ID: 12133412

To investigate the cause, prognosis and the treatment of fetal distress in pregnant women with hepatitis B virus (HBV) infection.

The Effect of Calmodulin Antagonist Berbaminederivative-EBB on Hepatoma in Vitro and in Vivo

Chinese Medical Journal. May, 2002  |  Pubmed ID: 12133551

To evaluate the anti-hepatoma effect of Calmodulin antagonist 0 - 4-ethoxyl-butyl-Berbamine (EBB), one of the berbamine derivatives.

[Intravenous Contrast Imaging in Diagnosis of Acute Renal Allograft Rejection: a Comparative Study with Color Flow Doppler Imaging]

Di 1 Jun Yi Da Xue Xue Bao = Academic Journal of the First Medical College of PLA. May, 2003  |  Pubmed ID: 12754123

To assess the value of intermittent contrast second harmonic imaging (SHI) incorporating acoustic densitometry in the diagnosis of acute renal allograft rejection in comparison with color Doppler flow imaging (CDFI) for determining the resistance index (RI).

The Relationship Between Serum Interleukins and T-lymphocyte Subsets in Patients with Severe Acute Respiratory Syndrome

Chinese Medical Journal. Jul, 2003  |  Pubmed ID: 12890366

To observe the changes of serum interleukins (IL), T-lymphocyte subsets, and white blood cell (WBC) count in patients with severe acute respiratory syndrome (SARS), and to investigate the relationship between injured immune function, immune response and disturbed immune adjustment in SARS patients.

[Detection of the Mutation in HBV Polymerase Gene by RFLP PCR Method in Hepatitis B Patients Treated with Lamivudine]

Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi = Zhonghua Shiyan He Linchuang Bingduxue Zazhi = Chinese Journal of Experimental and Clinical Virology. Sep, 2003  |  Pubmed ID: 15340574

To investigate the mutation of HBV polymerase gene in chronic hepatitis B patients treated with lamivudine.

Expression of SOCS-1, Suppressor of Cytokine Signalling-1, in Human Melanoma

The Journal of Investigative Dermatology. Oct, 2004  |  Pubmed ID: 15373779

Cytokine resistance is a well-established feature of melanoma cell progression and represents also a major obstacle in immunotherapy of patients with metastatic melanoma. To check whether suppressors of cytokine signalling (SOCS) play a role in cytokine resistance and tumor progression of melanoma, we investigated the expression and regulation of SOCS-1, an established negative regulator of interleukin-6 (IL-6) and interferon (IFN) signalling. In vitro SOCS-1 transcripts were detectable by RT-PCR in 8 out of 8 human melanoma cell lines derived from different tumor stages. Normal human melanocytes also expressed SOCS-1 mRNA in the presence or absence of artificial growth factors. Both IL-6 and alpha-IFN induced rapid and transient SOCS-1 mRNA expression in WM35 and WM9 melanoma cells. At the protein level, SOCS-1 was undetectable in normal human melanocytes whereas uniformly expressed in all tested melanoma cell lines. The aberrant SOCS-1 protein expression in melanoma cells was recapitalized in situ as shown by immunohistochemical analysis. SOCS-1 immunoreactivity was closely related to tumor invasion (Clark level), tumor thickness according to Breslow, and stage of the disease. In contrast, melanocytes in normal skin or melanocytic nevi lacked SOCS-1 protein expression. Our findings show that melanoma cells express a member of the SOCS family, SOCS-1, in vitro and in situ. SOCS-1 is a progression marker of human melanoma and may downregulate biological responses by endogenous and/or therapeutically administered cytokines.

[Clinical Significance of Detecting RNA and Anti HEV Antibody in Convalesent Sera in Patients with Acute HEV Hepatitis]

Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi = Zhonghua Shiyan He Linchuang Bingduxue Zazhi = Chinese Journal of Experimental and Clinical Virology. Sep, 2004  |  Pubmed ID: 15640865

To investigate the anti hepatitis E virus (HEV) and HEV RNA in acute and convalescent sera of patients with NonA-E acute hepatitis.

Electron-microscopic Structure of the V-ATPase from Mung Bean

Planta. Oct, 2004  |  Pubmed ID: 15185079

The vacuolar H(+)-ATPase from mung bean (Vigna radiata L. cv. Wilczek) was purified to homogeneity. The purified complex contained all the reported subunits from mung bean, but also included a 40-kDa subunit, corresponding to the membrane-associated subunit d, which has not previously been observed. The structure of the V-ATPase from mung bean was studied by electron microscopy of negatively stained samples. An analysis of over 6,000 single-particle images obtained by electron microscopy of the purified complex revealed that the complex, similar to other V-ATPases, is organized into two major domains V1 and Vo with overall dimensions of 25 nm x 13.7 nm and a stalk region connecting the V1 and Vo domains. Several individual areas of protein density were observed in the stalk region, indicating its complexity. The projections clearly showed that the complex contained one central stalk and at least two peripheral stalks. Subcomplexes containing subunits A, B and E, dissociated from the tonoplast membrane by KI, were purified. The structure of the subcomplex was also studied by electron microscopy followed by single-molecule analysis of 13,000 projections. Our preliminary results reveal an area of high protein density at the bottom of the subcomplex immediately below the cavity formed by the A and B subunits, indicating the position of subunit E.

Comparison of Nuclear Matrix Proteins Between Gastric Cancer and Normal Gastric Tissue

World Journal of Gastroenterology : WJG. Jun, 2004  |  Pubmed ID: 15188514

To study the alteration of nuclear matrix proteins (NMPs) in gastric cancer.

Demonstration of Interactions Among Myxococcus Xanthus Dif Chemotaxis-like Proteins by the Yeast Two-hybrid System

Archives of Microbiology. May, 2005  |  Pubmed ID: 15770502

The Myxococcus xanthus dif locus encodes several bacterial chemotaxis homologues that are crucial for fibril exopolysaccharide (EPS) production, social gliding motility, and fruiting body development. In primary sequence, DifA is homologous to methyl-accepting chemotaxis protein, DifC to CheW, DifD to CheY, DifE to CheA, and DifG to CheC. In this study, the interactions among the Dif chemotaxis-like proteins were investigated using the yeast two-hybrid (Y2H) system. DifC was found to interact with both DifA and DifE. Using a modified Y2H or a "three-hybrid" system, it was demonstrated that DifC is capable of mediating the formation of DifA, DifC, and DifE ternary protein complexes. The conserved domains of DifE, based on sequence analysis, likely reflect functional conservations of CheA-type kinases, because its P2 domain interacts with DifD, P5 with DifC, and the P3 domain appears to dimerize. Similarly, C-terminal regions of DifA appear to dimerize as well. In addition, DifG was found to interact with DifD, which is consistent with the hypothesis that DifG is a phosphatase of DifD-phosphate. These findings support the models in which Dif proteins constitute a unique chemotaxis-like signal transduction pathway with central functions in regulating EPS production in M. xanthus.

[Comparision of Several Different Peritoneal Fibrosis Rat Models]

Zhong Nan Da Xue Xue Bao. Yi Xue Ban = Journal of Central South University. Medical Sciences. Apr, 2005  |  Pubmed ID: 15898425

To compare different types of peritoneal fibrosis models in rats.

Improved Rapid Amplification of CDNA Ends (RACE) for Mapping Both the 5' and 3' Terminal Sequences of Paramyxovirus Genomes

Journal of Virological Methods. Dec, 2005  |  Pubmed ID: 16076500

Rapid amplification of cDNA ends (RACE) is a powerful PCR-based technique for determination of RNA terminal sequences. However, most of the RACE methods reported in the literature are developed specifically for the mapping of eukaryotic transcripts with 3' poly-A tail and 5' cap structure. In this study, an improved RACE strategy was developed which allows both 5' and 3' RACE of paramyxovirus genomic RNA using the same set of common molecular biology reagents without having to rely on expensive RACE kits. Mapping of RNA genome terminal sequences is an essential part of characterizing novel paramyxoviruses since these sequences contain important signals for genome replication and transcription, and are important molecular markers for studying virus evolution. The usefulness of this strategy was demonstrated by rapid characterization of both genome ends for a novel paramyxovirus recently isolated from human kidney primary cells. The RACE strategy described in this paper is simple, cost-effective and can be used to map genome ends of any RNA viruses.

Detection of Functionally Active Melanocortin Receptors and Evidence for an Immunoregulatory Activity of Alpha-melanocyte-stimulating Hormone in Human Dermal Papilla Cells

Endocrinology. Nov, 2005  |  Pubmed ID: 16081629

Proopiomelanocortin (POMC)-derived peptides and their receptors have been identified in many peripheral organs including the skin in which they exert a diversity of biological actions. We investigated the expression and potential role of the POMC system in human dermal papilla cells (DPCs), a specialized cutaneous mesenchymal cell type regulating hair follicle activity. In culture, these cells expressed POMC and displayed immunoreactivity for ACTH, alphaMSH, and beta-endorphin. Among the prohormone convertases (PCs) tested, only PC2, its chaperone 7B2, and furin convertase but not PC1 and paired basic amino acid cleaving enzyme 4 gene were detected. Human DPCs in vitro expressed both the melanocortin-1 receptor (MC-1R) and MC-4R, and immunoreactivity for these receptors was also present in cells of the human dermal papilla in situ. In contrast to the dermal papilla of agouti mice, agouti signaling protein, a natural and highly selective MC-1R and MC-4R antagonist, was undetectable in human DPCs. The MC-Rs detected in human DPCs were functionally active because alphaMSH increased intracellular cAMP and calcium. Preincubation of the cells with a synthetic peptide corresponding to the C-terminal domain of agouti signaling protein abrogated cAMP induction by alphaMSH. Furthermore, alphaMSH was capable of antagonizing the expression of intercellular adhesion molecule-1 induced by the proinflammatory cytokine interferon-gamma. Our data suggest a regulatory function of alphaMSH within the dermal papilla whose disruption may lead to deregulation of immune and inflammatory responses of the hair follicle, thereby possibly contributing to the development of inflammatory forms of alopecia.

[Association of TNFA Polymorphisms with the Outcomes of HBV Infection]

Zhonghua Yi Xue Yi Chuan Xue Za Zhi = Zhonghua Yixue Yichuanxue Zazhi = Chinese Journal of Medical Genetics. Aug, 2005  |  Pubmed ID: 16086278

To explore whether the TNFA promoter single nucleotide polymorphisms (SNPs) are associated with the outcomes of hepatitis B virus(HBV) infection in Chinese Han population.

The Dif Chemosensory Pathway is Directly Involved in Phosphatidylethanolamine Sensory Transduction in Myxococcus Xanthus

Molecular Microbiology. Sep, 2005  |  Pubmed ID: 16102016

Myxococcus xanthus cells glide on solid surfaces and are chemotactically stimulated by certain phosphatidylethanolamine species. The dif gene cluster consists of six genes, difABCDEG, five of which encode proteins homologous to known chemotaxis proteins. DifA and DifE are required for the biosynthesis of fibrils, an extracellular matrix comprised of polysaccharide and protein. Chemotactic stimulation by 1,2-O-Bis[11-(Z)-hexadecenoyl]-sn-glycero-3-phosphatidylethanolamine (16:1 PE) and dilauroyl PE (12:0 PE) requires fibrils. Although previous work has shown that difA and difE mutants are not stimulated by 12:0 PE, these results do not distinguish between a dependence on fibrils or a direct role in chemosensory transduction. Here we provide evidence that the Dif chemosensory pathway directly mediates PE sensory transduction. First, stimulation by and adaptation to 16:1 PE requires all of the dif genes, including difBDG, which are not essential for fibril biogenesis. Second, a specific residue within the first putative methylation domain of DifA is required for stimulation by 16:1 PE but not fibril biogenesis. Transmembrane signalling through a chimeric NarX-DifA chemoreceptor is required for fibril formation but not for stimulation by or adaptation to 16:1 PE. Third, difD and difE are required for stimulation by dioleoyl PE (18:1 PE) although the response does not require fibrils. Taken together these results argue that the Dif pathway mediates both matrix formation and lipid chemotaxis.

Association of Polymorphism of Tumor Necrosis Factor-alpha Gene Promoter Region with Outcome of Hepatitis B Virus Infection

World Journal of Gastroenterology : WJG. Sep, 2005  |  Pubmed ID: 16127755

To determine whether -238G/A and -857C/T polymorphisms of tumor necrosis factor-alpha (TNF-alpha), gene promoter and hepatitis B (HB) viral genotypes were associated with outcomes of HBV infection.

Beilong Virus, a Novel Paramyxovirus with the Largest Genome of Non-segmented Negative-stranded RNA Viruses

Virology. Mar, 2006  |  Pubmed ID: 16325221

During a subtraction study on gene expression in human kidney mesangial cells (HMCs), cDNA clones with sequence homology to paramyxovirus P, M and F genes were isolated. Subsequent investigation revealed that this particular HMC line was infected with a previously unknown paramyxovirus. Here, we report the isolation and genome characterization of this new virus, now named Beilong virus (BeV). The genome of BeV is 19,212 nucleotides (nt) in length and is the largest among all known members of the order Mononegavirales. The BeV genome contains eight genes in the order 3'-N-P/V/C-M-F-SH-TM-G-L-5'. The SH and TM genes code for a small hydrophobic protein of 76 aa and a transmembrane protein of 254 aa, respectively. The BeV G gene, at 4527 nt, codes for an attachment protein of 734 aa and contains two additional open reading frames (ORFs) in the 3' half of the gene, coding for putative proteins of 299 and 394 aa in length. Although the exact origin of BeV is presently unknown, we provide evidence indicating that BeV was present in a rat mesangial cell line used in the same laboratory prior to the acquisition of the HMC line, suggesting a potential rodent origin for BeV.

Magnetosomes Are Cell Membrane Invaginations Organized by the Actin-like Protein MamK

Science (New York, N.Y.). Jan, 2006  |  Pubmed ID: 16373532

Magnetosomes are membranous bacterial organelles sharing many features of eukaryotic organelles. Using electron cryotomography, we found that magnetosomes are invaginations of the cell membrane flanked by a network of cytoskeletal filaments. The filaments appeared to be composed of MamK, a homolog of the bacterial actin-like protein MreB, which formed filaments in vivo. In a mamK deletion strain, the magnetosome-associated cytoskeleton was absent and individual magnetosomes were no longer organized into chains. Thus, it seems that prokaryotes can use cytoskeletal filaments to position organelles within the cell.

Characterization of Solvated Electrons in Hydrogen Cyanide Clusters: (HCN)n- (n=3, 4)

The Journal of Chemical Physics. Feb, 2006  |  Pubmed ID: 16468872

Theoretical studies of the solvated electrons (HCN)n- (n=3, 4) reveal a variety of electron trapping possibilities in the (HCN)n (n=3, 4) clusters. Two isomers for (HCN)3- and four isomers for (HCN)4- are obtained at the MP2aug-cc-pVDZ+dBF (diffusive bond functions) level of theory. In view of vertical electron detachment energies (VDEs) at the CCSD(T) level, the excess electron always "prefers" locating in the center of the system, i.e., the isomer with higher coordination number shows larger VDE value. However, the most stable isomers of the solvated electron state (HCN)3- and (HCN)4- are found to be the linear Cinfinitynu and Dinfinityh structures, respectively, but not the fullyl symmetric structures which have the largest VDE values.

[Relationship Between Polymorphisms of Vitamin D Receptor Gene and Familial Aggregation of HBsAg Carriers]

Zhongguo Yi Xue Ke Xue Yuan Xue Bao. Acta Academiae Medicinae Sinicae. Apr, 2006  |  Pubmed ID: 16733893

To determine whether -Taq I T/C and -Fok I C/T polymorphisms of vitamin D receptor (VDR) gene are associated with the familial aggregation of hepatitis B virus (HBV) infection.

Association Of-238G/A and -857C/T Polymorphisms of Tumor Necrosis Factor-alpha Gene Promoter Region with Outcomes of Hepatitis B Virus Infection

Biomedical and Environmental Sciences : BES. Apr, 2006  |  Pubmed ID: 16827185

To determine whether -238G/A and -857C/T polymorphisms of tumor necrosis factor-alpha (TNF-alpha) gene promoter were associated with outcomes of hepatitis B virus infection.

[Study on Association Between Vitamin D Receptor Gene Polymorphisms and the Outcomes of HBV Infection]

Zhonghua Yi Xue Yi Chuan Xue Za Zhi = Zhonghua Yixue Yichuanxue Zazhi = Chinese Journal of Medical Genetics. Aug, 2006  |  Pubmed ID: 16883526

To explore whether the vitamin D receptor gene (VDR) polymorphisms are associated with the outcomes of hepatitis B virus (HBV) infection in Chinese Han population.

Cloning, Expression and Characterization of a Thermostable Exo-beta-D-glucosaminidase from the Hyperthermophilic Archaeon Pyrococcus Horikoshii

Biotechnology Letters. Oct, 2006  |  Pubmed ID: 16912928

An exo-beta-D-glucosaminidase gene (PH0511) was cloned from the hyperthermophilic archaeon, Pyrococcus horikoshii, and expressed in Escherichia coli. The purified protein showed a strong exo-beta-D: -glucosaminidase activity by TLC analysis. DTT (50 mM) had little effect on its homodimeric structure during SDS-PAGE. The enzyme was optimally active at 90 degrees C (over 20 min) and pH 6. It had a half-life of 9 h at 90 degrees C and is the most thermostable glucosaminidase described up to now. The activity was not inhibited by ethanol, 2-propanol, DMSO, PEG-400, denaturing agents SDS (5%, w/v), urea, guanidine hydrochloride (5 M) and Mg(2+), Mn(2+), Co(2+), Ca(2+), Sr(2+), Ni(2+) (at up to 10 mM).

[Effects of High Glucose on the Cell Proliferation, Damage and Cytokine in Human Peritoneal Mesothelial Cells]

Zhong Nan Da Xue Xue Bao. Yi Xue Ban = Journal of Central South University. Medical Sciences. Aug, 2006  |  Pubmed ID: 16951522

To determine the mechanism of peritoneal fibrosis of peritoneal mesothelial cells by high glucose.

Multiple Large Filament Bundles Observed in Caulobacter Crescentus by Electron Cryotomography

Molecular Microbiology. Oct, 2006  |  Pubmed ID: 16987173

While the absence of any cytoskeleton was once recognized as a distinguishing feature of prokaryotes, it is now clear that a number of different bacterial proteins do form filaments in vivo. Despite the critical roles these proteins play in cell shape, genome segregation and cell division, molecular mechanisms have remained obscure in part for lack of electron microscopy-resolution images where these filaments can be seen acting within their cellular context. Here, electron cryotomography was used to image the widely studied model prokaryote Caulobacter crescentus in an intact, near-native state, producing three-dimensional reconstructions of these cells with unprecedented clarity and fidelity. We observed many instances of large filament bundles in various locations throughout the cell and at different stages of the cell cycle. The bundles appear to fall into four major classes based on shape and location, referred to here as 'inner curvature', 'cytoplasmic', 'polar' and 'ring-like'. In an attempt to identify at least some of the filaments, we imaged cells where crescentin and MreB filaments would not be present. The inner curvature and cytoplasmic bundles persisted, which together with their localization patterns, suggest that they are composed of as-yet unidentified cytoskeletal proteins. Thus bacterial filaments are frequently found as bundles, and their variety and abundance is greater than previously suspected.

Detection of HEV Antigen As a Novel Marker for the Diagnosis of Hepatitis E

Journal of Medical Virology. Nov, 2006  |  Pubmed ID: 16998897

Infection with hepatitis E virus (HEV) may be diagnosed by the presence of HEV RNA or anti-HEV antibodies. An enzyme immunoassay (EIA) was developed for the detection of antigen. Twenty-four monoclonal antibodies (mAbs) were produced. An indirect sandwich EIA was developed to detect HEV antigen using a combination of three mAbs as coating antibodies. Approximately 44.6% (33/74), 28.6% (50/175), and none (0/27) of sera positive for anti-HEV IgM alone, both anti-HEV IgM and IgG, and anti-HEV IgG alone also were positive for HEV antigen using this EIA. Forty-two HEV antibody-positive sera were tested for HEV RNA and antigen in parallel and the concordance was 81.0% (34/42). All PCR products were found to belong to HEV genotype 4. In order to evaluate the temporal relationship between HEV antigen positivity and HEV RNA, anti-HEV IgG and IgM, and ALT concentrations, macaques were infected with HEV genotypes 1 and 4 and serial samples were collected. The results showed that the antigen EIA can detect the capsid proteins of both genotypes. HEV antigen was detectable prior to ALT elevation and the appearance of anti-HEV antibodies in the infected monkeys and lasted for several weeks in all cases. HEV antigen became detectable in the serum at almost the same time as HEV RNA in feces but persisted for 4 weeks less than HEV RNA. This assay should be valuable for the diagnosis of acute hepatitis E, particularly in the window period prior to seroconversion to anti-HEV.

Construction of Methylation-sensitive Partial Restriction Bacterial Artificial Chromosome Libraries in Maize

Analytical Biochemistry. Dec, 2006  |  Pubmed ID: 17010300

[Association of Taq I T/C and Fok I C/T Polymorphisms of Vitamin D Receptor Gene with Outcome of Hepatitis B Virus Infection]

Zhonghua Yi Xue Za Zhi. Jul, 2006  |  Pubmed ID: 17064588

To determine whether Taq I T/C and Fok I C/T polymorphisms of vitamin D Receptor (VDR) gene was associated with the outcomes of hepatitis B virus (HBV) infection.

Electron Cryotomography Sample Preparation Using the Vitrobot

Nature Protocols. 2006  |  Pubmed ID: 17406539

Electron cryotomography is the highest-resolution structural technique currently available that can be applied to unique objects such as flexible large protein complexes, irregular viruses, organelles and small cells. Specimens are preserved in a near-native, 'frozen-hydrated' state by vitrification. The thickness of the vitreous ice must be optimized for each specimen, and gold fiducials are typically added to facilitate image alignment. Here, we describe in detail our protocols for electron cryotomography sample preparation including (i) introduction of fiducial markers into the sample and (ii) sample vitrification. Because we almost exclusively use an automated, climate-controlled plunge-freezing device (the FEI Vitrobot) to vitrify our samples, we discuss its operation and parameters in detail. A session in which eight grids are prepared takes 1.5-2 h.

[Genetic Polymorphisms of MxA Protein and EIF-2a-reg2 and Their Responses to Interferon Treatment in Patients with Chronic Hepatitis B]

Zhonghua Gan Zang Bing Za Zhi = Zhonghua Ganzangbing Zazhi = Chinese Journal of Hepatology. Mar, 2007  |  Pubmed ID: 17407708

To identify the host single nucleotide polymorphisms (SNP) of myxovirus resistance A (MxA) protein and eukaryote initiation factor 2alfa regulatory region 2(eIF-2a-reg2) and to predict interferon (IFN) treatment responses in patients with chronic hepatitis B.

Chemoselective C(sp3)-H Bond Activation for the Preparation of Condensed N-heterocycles

Chemistry, an Asian Journal. Mar, 2007  |  Pubmed ID: 17441179

Condensed N-heterocycles were prepared by using C-H activation reactions catalyzed by Pd(OAc)2 (5 mol %) and (p-tolyl)3P (10 mol %). The key step of these ring closures is chemoselective intramolecular C-H activation of the methyl group at position 2 of the pyrrole ring. Functionalized 9H-pyrrolo[1,2-a]indoles and pyrrolo[1,2-f]phenanthridine derivatives were prepared in good yields. The preparation of some complex N-heterocycles by using successive reactions is also described.

A Unique Highly Thermostable 2-phosphoglycerate Forming Glycerate Kinase from the Hyperthermophilic Archaeon Pyrococcus Horikoshii: Gene Cloning, Expression and Characterization

Extremophiles : Life Under Extreme Conditions. Sep, 2007  |  Pubmed ID: 17563835

A glycerate kinase (GK) gene (PH0495) from the hyperthermophilic archaeon Pyrococcus horikoshii, was cloned and expressed in Escherichia coli. The recombinant protein was purified to homogeneity by affinity chromatography and ion exchange chromatography. The enzyme was likely a homodimer based on SDS-PAGE (47 kDa) and gel filtration chromatography (100 kDa) analysis. A radioisotope-labeling examination method was initially used for the enzymatic activity detection, and the enzyme (GK(ph)) was found to catalyze the formation of 2-phosphoglycerate using D: -glycerate as the substrate. The enzyme exhibited unique phosphoryl donor specificity with maximal activity towards pyrophosphate. The temperature and pH optima of the enzyme were 45 degrees C and 7.0, respectively, and about half of the maximal activity remained at 100 degrees C. The enzyme was highly thermostable with almost no loss of activity at 90 degrees C for 12 h. Based on sequence alignment and structural comparison it was assigned to group I of the trichotomy of GKs.

[Stressed Level of Urban Vegetation: Its Assessment Based on Hyperion Hyperspectral Data]

Ying Yong Sheng Tai Xue Bao = The Journal of Applied Ecology / Zhongguo Sheng Tai Xue Xue Hui, Zhongguo Ke Xue Yuan Shenyang Ying Yong Sheng Tai Yan Jiu Suo Zhu Ban. Jun, 2007  |  Pubmed ID: 17763731

To quickly obtain the information of urban vegetation stressed level is of great significance in maintaining urban vegetation health and improving urban eco-environment. Based on the analysis of stressed vegetations physiological and spectral characters, and by using Hyperion hyperspectral data, 14 hyperspectral vegetation indices related to stress were calculated, and a classifier of urban vegetation stressed level was developed based on this calculation and BP Neural network. The application of this classifier in identifying the vegetation stressed level in a case study area of Guangzhou City showed that the vegetations in commercial and residential districts were apparently experienced higher stress than those in suburban regions, and the stressed level showed a ringy distribution around large pieces of greenbelts. This classifier was able to quickly and accurately identify the vegetation stressed level, and thus, could be used as an effective tool in monitoring urban vegetation stressed condition.

Haloquadratum Walsbyi Gen. Nov., Sp. Nov., the Square Haloarchaeon of Walsby, Isolated from Saltern Crystallizers in Australia and Spain

International Journal of Systematic and Evolutionary Microbiology. Feb, 2007  |  Pubmed ID: 17267984

Strains C23T and HBSQ001 were isolated from solar salterns and are novel square-shaped, aerobic, extremely halophilic members of the domain Archaea and family Halobacteriaceae. Cells stained Gram-negative and grew optimally in media containing 18 % salts at around neutral pH. Mg2+ is not required. The DNA G+C content of both isolates was 46.9 mol% and DNA-DNA cross-hybridization showed a relatedness of 80 %. Their 16S rRNA gene sequences showed only 2 nucleotide differences (99.9 % identity) and phylogenetic tree reconstructions with other recognized members of the Halobacteriaceae indicated that they formed a distinct clade, with the closest relative being Halogeometricum borinquense PR 3T (91.2 % sequence identity). The major polar glycolipid of both isolates was the sulfated diglycosyl diether lipid S-DGD-1. Electron cryomicrosopy of whole cells revealed similar internal structures, such as gas vesicles and polyhydroxyalkanoate granules, but the cell wall of isolate HBSQ001 displayed a more complex S-layer compared with that of isolate C23T. The phenotypic characterization and phylogenetic data support the placement of isolates C23T and HBSQ001 in a novel species in a new genus within the Halobacteriaceae, for which we propose the name Haloquadratum walsbyi gen. nov., sp. nov. The type strain of Haloquadratum walsbyi is C23T (=JCM 12705T=DSM 16854T).

Association of HLA-DQB1 Gene Polymorphisms with Outcomes of HBV Infection in Chinese Han Population

Swiss Medical Weekly. Feb, 2007  |  Pubmed ID: 17370149

Host genetic factors and environmental factors including hepatitis B virus (HBV) genotype are widely studied for the different outcomes of HBV infection. Human leukocyte antigen (HLA) plays an important role in the immunological reaction to HBV infection.

Comparison of Real-time Fluorescent RT-PCR and Conventional RT-PCR for the Detection of Hepatitis E Virus Genotypes Prevalent in China

Journal of Medical Virology. Dec, 2007  |  Pubmed ID: 17935186

To compare the specificity and sensitivity of a real-time fluorescent RT-PCR assay with conventional RT-PCR, sera from 110 healthy blood donors, 120 patients with a clinical diagnosis of chronic hepatitis B, and 416 patients with non-A-C acute hepatitis, as well as serial dilutions of HEV genotypes 1 and 4, were tested with both assays. All samples from healthy blood donors and patients with chronic hepatitis B were negative by both assays. Real-time RT-PCR could detect the same final dilution of genotype 1 as conventional RT-PCR but could detect a 10-fold lower concentration of genotype 4 than conventional RT-PCR. Of 416 samples from patients with a clinical diagnosis of non-A-C acute hepatitis, 127 (30.5%) and 83 (20.0%) were positive for HEV by real-time and conventional RT-PCR, respectively. The concordance of real-time and conventional RT-PCR was 80.8%. Furthermore, 96 and 57 of 171 samples were positive for anti-HEV IgM by real-time and conventional RT-PCR, respectively, and 31 and 26 of 245 samples negative for anti-HEV IgM, were positive by real-time and conventional RT-PCR, respectively. All amplicons positive by conventional RT-PCR were sequenced. Of 83 isolates, 7 and 76 belonged to genotypes 1 and 4, respectively. Thus, both assays have a high specificity, but the real-time RT-PCR assay is more sensitive than conventional RT-PCR. Furthermore, HEV genotype 4 is responsible for most sporadic cases of hepatitis E in the north of China.

The Structure of FtsZ Filaments in Vivo Suggests a Force-generating Role in Cell Division

The EMBO Journal. Nov, 2007  |  Pubmed ID: 17948052

In prokaryotes, FtsZ (the filamentous temperature sensitive protein Z) is a nearly ubiquitous GTPase that localizes in a ring at the leading edge of constricting plasma membranes during cell division. Here we report electron cryotomographic reconstructions of dividing Caulobacter crescentus cells wherein individual arc-like filaments were resolved just underneath the inner membrane at constriction sites. The filaments' position, orientation, time of appearance, and resistance to A22 all suggested that they were FtsZ. Predictable changes in the number, length, and distribution of filaments in cells where the expression levels and stability of FtsZ were altered supported that conclusion. In contrast to the thick, closed-ring-like structure suggested by fluorescence light microscopy, throughout the constriction process the Z-ring was seen here to consist of just a few short (approximately 100 nm) filaments spaced erratically near the division site. Additional densities connecting filaments to the cell wall, occasional straight segments, and abrupt kinks were also seen. An 'iterative pinching' model is proposed wherein FtsZ itself generates the force that constricts the membrane in a GTP-hydrolysis-driven cycle of polymerization, membrane attachment, conformational change, depolymerization, and nucleotide exchange.

[Association Between Genetic Polymorphism of Tumor Necrosis Factor and Chronic Severe Hepatitis B in Patients]

Zhonghua Yi Xue Za Zhi. Aug, 2007  |  Pubmed ID: 17988528

To investigate the association between the single nucleotide polymorphisms (SNPs) of tumor necrosis factor (TNF-alpha) and chronic severe hepatitis B.

Tricuspid Regurgitation in Patients Undergoing Pericardiectomy for Constrictive Pericarditis

The Annals of Thoracic Surgery. Jan, 2008  |  Pubmed ID: 18154802

Tricuspid regurgitation (TR) may complicate pericardial constriction; however, its incidence, impact on outcome, and appropriate management are not defined.

Hjm/Hel308A DNA Helicase from Sulfolobus Tokodaii Promotes Replication Fork Regression and Interacts with Hjc Endonuclease in Vitro

Journal of Bacteriology. Apr, 2008  |  Pubmed ID: 18296528

Hjm and Hel308a are novel, RecQ-like DNA helicases recently identified in the euryarchaeotes Pyrococcus furiosus and Methanothermobacter thermautotrophicus, respectively. In this study, an Hjm/Hel308 homologue (designated StoHjm) from Sulfolobus tokodaii, a hyperthermophilic archaeon belonging to the Crenarchaeota subdomain of archaea, was cloned, purified, and characterized. Unlike Hjm and Hel308a, which unwind DNA in a 3'-to-5' direction, StoHjm unwound DNA in both 3'-to-5' and 5'-to-3' directions. Remarkably, StoHjm exhibited structure-specific single-stranded-DNA-annealing and fork regression activities in vitro. In addition, gel filtration, affinity pulldown, and yeast two-hybrid analyses revealed that StoHjm physically interacted with StoHjc, the Holliday junction-specific endonuclease from S. tokodaii. This interaction may have functional significance, because the unwinding activity of StoHjm was inhibited by StoHjc in vitro. These results may suggest that the Hjm/Hel308 family helicases, in association with Hjc endonucleases, are involved in processing of stalled replication forks.

Hepatocyte Growth Factor Combined with Insulin Like Growth Factor-1 Improves Expression of GATA-4 in Mesenchymal Stem Cells Cocultured with Cardiomyocytes

Chinese Medical Journal. Feb, 2008  |  Pubmed ID: 18304467

In a suitable microenvironment, bone marrow mesenchymal stem cells (BMSCs) can transdifferentiate into myocardial cells whose special gene can be expressed as structural proteins. Growth factor (GF) plays an important role in the cell migration, survival and differentiation. However, the effect of GF on the cellular differentiation is not well understood. In this study, the hepatocyte growth factor (HGF) and insulin like growth factor-1 (IGF-1) were used in the mixed culture of BMSCs and myocardial cells and the effects of these growth factors on the GATA-4 expression of BMSCs were investigated.

Modeling Motor Vehicle Crashes for Street Racers Using Zero-inflated Models

Accident; Analysis and Prevention. Mar, 2008  |  Pubmed ID: 18329439

Motor vehicle crashes are a leading cause of death for young people in the United States. Assessing which drivers are at a high risk of experiencing a crash is important for the implementation of traffic regulations. Illegal street racing has been associated with a high rate of motor vehicle crashes. In this study, we link Utah statewide driver license citations and motor vehicle crash data to evaluate the rate of crashes for drivers with street racing citations relative to other drivers. Using a zero-inflated negative binomial model we found that drivers with no citations are approximately three times more likely to be at zero risk of a crash compared to drivers with street racing citations. Moreover, among drivers at non-negligible risk of crash, cited street racers are more likely to be involved in a crash compared to drivers without citations or those cited for violations other than street racing. However, drivers with increased numbers of non-street-racing citations experience crash risks approaching those of the cited street racers.

The Novel Gene AngRem104 Downregulates Glucocorticoid Receptor Expression and Activates NF-kappaB in Human Mesangial Cells

Biochemical and Biophysical Research Communications. May, 2008  |  Pubmed ID: 18331827

AngRem104 [angiotensin II (Ang II)-related genes in human mesangial cells (MCs), clone104], a novel gene in human MCs induced by Ang II, was previously identified in human MCs and found to interact with several proteins. The current study used a yeast two-hybrid system and co-immunoprecipitation to investigate the interaction between AngRem104 and glucocorticoid receptor (GR) AF-1-specific elongation factor (GR-EF). GR expression was downregulated and the number of MCs positive for activated nuclear factor kappaB (NF-kappaB) was increased when AngRem104 was overexpressed. Transfection with antisense AngRem104 vector resulted in the upregulation of GR protein and reduced numbers of MCs with activated NF-kappaB. These results indicate that the novel gene AngRem104 is involved in the in vivo regulation of GR expression and the activation of NF-kappaB through interaction with GR-EF in human MCs.

Location and Architecture of the Caulobacter Crescentus Chemoreceptor Array

Molecular Microbiology. Jul, 2008  |  Pubmed ID: 18363791

A new method for recording both fluorescence and cryo-EM images of small bacterial cells was developed and used to identify chemoreceptor arrays in cryotomograms of intact Caulobacter crescentus cells. We show that in wild-type cells preserved in a near-native state, the chemoreceptors are hexagonally packed with a lattice spacing of 12 nm, just a few tens of nanometers away from the flagellar motor that they control. The arrays were always found on the convex side of the cell, further demonstrating that Caulobacter cells maintain dorsal/ventral as well as anterior/posterior asymmetry. Placing the known crystal structure of a trimer of receptor dimers at each vertex of the lattice accounts well for the density and agrees with other constraints. Based on this model for the arrangement of receptors, there are between one and two thousand receptors per array.

The Outcomes of Operations for 539 Patients with Ebstein Anomaly

The Journal of Thoracic and Cardiovascular Surgery. May, 2008  |  Pubmed ID: 18455593

Our objective was to review the long-term outcomes of patients with Ebstein anomaly who underwent cardiac surgery at our institution.

Analytical Design of Conformally Invisible Cloaks for Arbitrarily Shaped Objects

Physical Review. E, Statistical, Nonlinear, and Soft Matter Physics. Jun, 2008  |  Pubmed ID: 18643390

To design conformally invisible cloaks for arbitrarily shaped objects, we use the nonuniform rational B -spline (NURBS) to represent the geometrical modeling of the arbitrary object. Based on the method of optical transformation, analytical formulas of the permittivity and permeability tensors are proposed for arbitrarily shaped invisible cloaks. Such formulas can be easily implemented in the design of arbitrary cloaks. Full-wave simulations are given for heart-shaped invisible cloaks and perfectly electrical conducting (PEC) objects, in which we observe that the power-flow lines of incoming electromagnetic waves will be bent smoothly in the cloaks and will return to their original propagation directions after propagating around the object. We also show that the scattered field from the PEC object coated with the invisible cloak is much smaller than that from the PEC core. The application of NURBS in the design of arbitrary cloaks shows transformation optics to be a very general tool to interface with commercial softwares like 3D STUDIOMAX and MAYA.

Incidence, Prevalence, and Management of Intraductal Papillary Mucinous Neoplasm in Olmsted County, Minnesota, 1984-2005: a Population Study

Pancreas. Aug, 2008  |  Pubmed ID: 18665073

Intraductal mucinous papillary neoplasm (IPMN) is being recognized with increasing frequency around the world. The true incidence, however, remains unknown. Our goal was to determine the incidence of IPMN in a population study.

Functional Status After Operation for Ebstein Anomaly: the Mayo Clinic Experience

Journal of the American College of Cardiology. Aug, 2008  |  Pubmed ID: 18672167

The objective of this study was to review the long-term functional outcome of patients with Ebstein anomaly who had cardiac operation at our institution.

Spatial Subunit Distribution and in Vitro Functions of the Novel Trimeric PCNA Complex from Sulfolobus Tokodaii

Biochemical and Biophysical Research Communications. Nov, 2008  |  Pubmed ID: 18782564

The relationships among three PCNA (proliferating cell nuclear antigen) subunits in the hyperthermophilic archaeon Sulfolobus tokodaii (StoPCNAs) were analyzed and the effects of two PCNA complexes on the activities of the DNA helicase Hjm, DNA Ligase I, and Holliday junction specific endonuclease Hjc were tested. There was no strong self-interaction of each StoPCNA. StoPCNA1 and StoPCNA3 interacted with each other, so did StoPCNA2 and StoPCNA3, but no interaction between StoPCNA1 and StoPCNA2 was observed. Two trimeric complexes (designed StoPCNA123 and StoPCNA323) were formed in vitro and it was determined that StoPCNA323 was composed of one StoPCNA2 and two StoPCNA3 subunits, with StoPCNA2 bridging the two StoPCNA3 subunits. Both complexes inhibited the unwinding activity of Hjm and the ligation activity of DNA Ligase I. In contrast, both stimulated the Holliday junction cleavage activity of Hjc. Our results provide further evidence that in crenarchaea, the PCNAs exhibit diversity in subunit interaction and complex formation.

Is the European System for Cardiac Operative Risk Evaluation Model Valid for Estimating the Operative Risk of Patients Considered for Percutaneous Aortic Valve Replacement?

The Journal of Thoracic and Cardiovascular Surgery. Sep, 2008  |  Pubmed ID: 18805253

The European System for Cardiac Operative Risk Evaluation has been used to define a particularly high-risk group of patients for aortic valve replacement in whom alternative procedures, such as stent-mounted percutaneous valve procedures, may be appropriate. Our objective was to assess the validity of this risk assessment at a large-volume, tertiary cardiac surgical center.

Alternate Deposition of Oriented Calcite and Amino Acid Layer on Calcite Substrates

The Journal of Physical Chemistry. B. Oct, 2008  |  Pubmed ID: 18841888

Material synthesis inspired by novel nacre architecture and mechanism is popular and has attracted more and more attention. In this paper, iso-oriented calcite tablets/layers and amino acid layers were formed alternately on calcite wafers. It is interesting that the neonatal calcite tablets/layers have the same crystal orientation with their inorganic substrates through amino acid layers. It is quite possible that the amino acid layers in this study could transfer crystal orientation from formed inorganic layers to neighboring neonatal layers due to their fixed and appropriate structures, which may imply the process of nacre formation, and the role of aligned organic matrix sheets in nacre. Moreover, it could provide a new way to produce oriented calcite tablets/layers.

The Diploid Genome Sequence of an Asian Individual

Nature. Nov, 2008  |  Pubmed ID: 18987735

Here we present the first diploid genome sequence of an Asian individual. The genome was sequenced to 36-fold average coverage using massively parallel sequencing technology. We aligned the short reads onto the NCBI human reference genome to 99.97% coverage, and guided by the reference genome, we used uniquely mapped reads to assemble a high-quality consensus sequence for 92% of the Asian individual's genome. We identified approximately 3 million single-nucleotide polymorphisms (SNPs) inside this region, of which 13.6% were not in the dbSNP database. Genotyping analysis showed that SNP identification had high accuracy and consistency, indicating the high sequence quality of this assembly. We also carried out heterozygote phasing and haplotype prediction against HapMap CHB and JPT haplotypes (Chinese and Japanese, respectively), sequence comparison with the two available individual genomes (J. D. Watson and J. C. Venter), and structural variation identification. These variations were considered for their potential biological impact. Our sequence data and analyses demonstrate the potential usefulness of next-generation sequencing technologies for personal genomics.

Vertical Columnar Block-copolymer-templated Mesoporous Silica Via Confined Phase Transformation

Journal of the American Chemical Society. Dec, 2008  |  Pubmed ID: 19053489

An efficient method is described for the preparation of phase-pure columnar mesoporous silica nanosystems within the channels of anodic alumina membranes (AAM) via evaporation-induced self-assembly (EISA). Upon the basis of a systematic investigation of the effects of interfacial interactions and different synthesis parameters on the resulting hierarchical mesophase, a salt-induced phase transformation was developed for efficient structural control. Samples with a columnar hexagonal 2D structure along the vertical channels of the AAM can be produced with ionic CTAB as template. However, when nonionic surfactants (Pluronic P123 and Brij 56) are used, samples with a circular hexagonal 2D structure perpendicular to the channels or phase mixtures of circular and columnar orientations are obtained. The behavior of ionic CTAB can be mimicked by adding inorganic salt to the nonionic template precursor solution, thus leading to a phase transformation toward columnar orientation. The distribution between the orientations was determined by means of small-angle X-ray scattering (SAXS) experiments. The effects of other synthesis parameters were also investigated, including temperature, surfactant: silica ratio, and salt composition. Strikingly, calcination-stable mesoporous materials with a columnar orientation exhibiting high mesoporosity and specific surface area were obtained for the first time with such structure directors. The salt-induced phase transformation is an efficient means for achieving a desired hierarchical mesostructure in the confined space of the AAM channels.

Effect of Operation for Ebstein Anomaly on Left Ventricular Function

The American Journal of Cardiology. Dec, 2008  |  Pubmed ID: 19064031

Our objective was to examine the outcomes of patients with left ventricular (LV) dysfunction who underwent operation for Ebstein anomaly. From April 1, 1972 to January 1, 2006, 539 patients with Ebstein anomaly underwent operation at Mayo Clinic. LV function was determined by echocardiography. Of the 495 patients with preoperative echocardiographic assessment of LV function, 50 had moderate or severe LV systolic dysfunction. In patients with LV dysfunction, the tricuspid valve (TV) was repaired in 12 patients and replaced in 36 patients; 1 patient had a 1.5 ventricle repair, and 1 patient had cardiac transplantation. There were 5 early deaths (10%). LV function improved in all but 4 patients after operation. In no patient did LV function worsen after operation. The 1-, 5-, and 10-year survival was 86%, 77%, and 67%, respectively. On univariate analysis, absence of sinus rhythm at dismissal (p = 0.003) was associated with greater overall mortality. For the entire cohort of 539 patients, LV dysfunction was independently predictive of late mortality (hazard ratio 3.76, p <0.001). At late follow-up (mean 6.9 years), 86% of patients were in New York Heart Association class I or II. In conclusion, LV systolic dysfunction occurs infrequently in patients with Ebstein anomaly and is a risk factor for increased late mortality. Although early mortality is greater in patients with LV dysfunction, the late results are favorable. Decreasing LV function should be an indication to promptly restore TV competence rather than a contraindication to TV operation.

Theoretical Elucidation of the Rhodium-catalyzed [4 + 2] Annulation Reactions

Journal of Computational Chemistry. Apr, 2008  |  Pubmed ID: 17849391

The reaction mechanism of the Rh-catalyzed [4 + 2] annulation of 4-alkynals with isocyanates is unraveled using density functional calculations. The reaction mechanisms of the model system and the real substituted system have been investigated and the results are compared. From our theoretical results based on the model and real substituted system, it is shown that (a) the rate-determining step is the Rh-H addition to the alkyne, (b) the formation of the cyclopentenone G and glutarimide K represents a severe competition, and (c) the product selectivity should be controlled by the amount of the isocyanates. In addition, it is demonstrated that there exist steric effects in the real substituted system, but missed in model system. Our calculations also show that although the results obtained on the model system could explain the mechanism in principle, the real substituted system could reflect the mechanism more exactly and make the reaction proceed with regioselectivity.

Isolation and Characterization of Phenanthrene-degrading Strains Sphingomonas Sp. ZP1 and Tistrella Sp. ZP5

Journal of Hazardous Materials. Apr, 2008  |  Pubmed ID: 17850962

Two bacteria strains Sphingomonas sp. strain ZP1 and Tistrella sp. strain ZP5 were identified as phenanthrene-degrading ones, based on Gram staining, oxydase reaction, biochemical tests, FAME analysis, G+C content and 16S rDNA gene sequence analysis. We isolated these two bacteria strains Sphingomonas sp. ZP1 and Tistrella sp. ZP5 from soil samples contaminated with polycyclic aromatic hydrocarbon (PAH)-containing waste from oil refinery field in Shanghai, China. Strain Sphingomonas sp. ZP1 was able to degrade naphthalene, phenanthrene, toluene, methanol and ethanol, salicylic acid and Tween 80. Moreover, it can remove nearly all the phenanthrene at 0.025% concentration in 8 days. Strain Tistrella sp. ZP5 cannot degrade phenanthrene individually but it can increase the speed of phenanthrene degradation together with ZP1. The growth conditions of strain Sphingomonas sp. ZP1 were optimized. The result also indicated that the degradation rate of phenanthrene ranged from 250 to 1000 ppm with strain ZP1 remained nearly the same, i.e., a high concentration of phenanthrene did not inhibit both the growth of microbial strains and the phenanthrene-degradation ability. Besides, the effect of non-ionic surfactants such as Brij 30, Triton X-100 and Tween 80 on the phenanthrene degradation was determined. Such two strains may be useful for bioremediation applications.

Cloning and Sequencing of V-ATPase Subunit D from Mung Bean and Its Function in Passive Proton Transport

Journal of Bioenergetics and Biomembranes. Dec, 2008  |  Pubmed ID: 19194790

We have previously shown that vacuolar H+-ATPase subcomplex V(o) from mung bean contains subunit d, however, its sequence and function were unknown. In the present study, we report the cloning and recombinant over expression of subunit d from mung bean in E. coli. To study the function of subunit d, two vacuolar H+-ATPase subcomplexes V(o) from mung bean were purified-one containing subunits a and c(c',c") and the other containing subunits a, c(c',c") and d. After reconstitution of the purified V(o) subcomplexes into liposomes, the proton translocation was studied. Our results show that the V(o) subcomplex in the absence of subunit d is a passive proton channel, while the V(o) subcomplex in the presence of the subunit d is not. Taken together, our data supports the conclusion that the subunit d of the plant vacuolar H(+)-ATPase from mung bean is positioned at the central stalk and involved in the proton translocation across the tonoplast membrane.

Comparison of the Outcome of Porcine Bioprosthetic Versus Mechanical Prosthetic Replacement of the Tricuspid Valve in the Ebstein Anomaly

The American Journal of Cardiology. Feb, 2009  |  Pubmed ID: 19195520

Our objective was to determine the relative merits of using a bioprosthetic porcine valve (BPV) versus a mechanical valve (MechV) when tricuspid valve (TV) replacement is required in patients with Ebstein anomaly. From 1972 to 2006, 333 patients received a BPV and 45 received a MechV. Patient records were reviewed, vital status ascertained, and all patients not known to be deceased were mailed a medical questionnaire or contacted by telephone. Early mortality was not statistically higher for patients who had a MechV (11%) than for those who had a BPV (5%) inserted in the TV position (p = 0.173). The only independent preoperative predictor of operative mortality was moderate to severe left ventricular dysfunction (odds ratio 3.1, p = 0.03); 20-year survival was better in patients who had a BPV (75%) than for those who had a MechV (43%, p = 0.003). On multivariate analysis, after adjusting for ablation of accessory pathways, sinus rhythm at dismissal, and concomitant repair of pulmonary valve stenosis, a BPV remained a predictor of late survival (hazard ratio 0.42, p = 0.004). Survival free of reoperation on the TV at 20 years postoperatively was similar for patients who had a MechV (49%) compared with those who had a BPV (42%) inserted (p = 0.941). A greater percentage of patients who had a MechV reported endocarditis (12% vs 2%), bleeding requiring hospitalization (6% vs 3%), and thrombosis (12% vs 6%); however, none of these differences were statistically significant. In conclusion, a BPV in the tricuspid position was an independent predictor of improved survival. This may be related to the higher incidence of bleeding and thrombotic complications in the patients with MechVs or may be related to differences between the 2 groups. A BPV may offer superior late survival when compared with a MechV when TV replacement is required in patients with Ebstein anomaly, but patient selection must be individualized.

Management of Moderate Functional Mitral Regurgitation at the Time of Aortic Valve Replacement: is Concomitant Mitral Valve Repair Necessary?

The Journal of Thoracic and Cardiovascular Surgery. Mar, 2009  |  Pubmed ID: 19258080

The optimal management of moderate functional mitral regurgitation at the time of aortic valve replacement remains undefined.

Indexed Left Ventricular Dimensions Best Predict Survival After Aortic Valve Replacement in Patients with Aortic Valve Regurgitation

The Annals of Thoracic Surgery. Apr, 2009  |  Pubmed ID: 19324145

Indications for valve replacement in patients with aortic regurgitation include diminished ejection fraction and increased left ventricular dimensions. Our objective was to examine the effect of preoperative ejection fraction and left ventricular dimensions on survival and return of normal systolic function (ejection fraction > or = 0.50) after valve replacement for aortic regurgitation.

[Basic Experimental and Clinical Research on Peritoneal Dialysis in the Past 16 Years]

Zhong Nan Da Xue Xue Bao. Yi Xue Ban = Journal of Central South University. Medical Sciences. Mar, 2009  |  Pubmed ID: 19349685

To summarized the experiences from our basic experimental and clinical research on peritoneal dialysis. In the past 16 years, peritoneal fibrosis rat models and rabbit models of peritonitis were first established successfully in our laboratory in China. Peritoneal mesothelial cells were also separated and identificated. Besides, we assessed the biocompatibility of peritoneal dialysis fluid and analyzed the molecular mechanism of peritoneal mesothelial cell injury. We demonstrated the key role of transforming growth factor-beta1 (TGF-beta1), connective tissue growth factor (CTGF) and peroxisome proliferative activated receptor-gamma (PPAR-gamma) in the pathogenesis of peritoneal fibrosis, as well as their regulation of molecular mechanism. Furthermore, we transfected the plasmids encoding TGF-beta1-shRNA or pCTGF-shRNA into peritoneal cells and tissues by nanocarrier technologies. In clinical research, the positioning of peritoneal dialysis catheters, peritoneal dialysis treatment modalities and the prevention and treatment of its complications were studied. The characteristics and mechanism of solute transport in peritoneal dialysis was also explored.

Varying Abilities of Recombinant Polypeptides from Different Regions of Hepatitis E Virus ORF2 and ORF3 to Detect Anti-HEV Immunoglobulin M

Journal of Medical Virology. Jun, 2009  |  Pubmed ID: 19382255

Following infection with hepatitis E virus (HEV), anti-HEV immunoglobulin (Ig) M is thought to develop before anti-HEV IgG and to be a better marker for differentiating between the acute and convalescent phases of infection. In order to select polypeptides for improved detection of anti-HEV IgM, six and three overlapping polypeptides from open reading frames (ORFs) 2 and 3, respectively, of HEV genotypes 1 and 4 were expressed as fusion proteins in Escherichia coli. The reactivities of the polypeptides with anti-HEV IgM were evaluated using immunoblotting and enzyme immunoassays (EIAs). The data indicated that polypeptides from the N-terminus of ORF3 and middle region of ORF2 were weakly or not reactive with anti-HEV IgM, while those from the remaining regions of ORF2 and ORF3 contained reactive epitopes. Anti-HEV IgM against the N- or C-terminus of ORF2 appeared earlier and disappeared faster than that against polypeptides from the C-terminus of ORF3, based on serum samples from rhesus monkeys infected experimentally, and from patients infected naturally, with HEV. The N- and C-terminal polypeptides from ORF2 complemented one another in detecting anti-HEV IgM and EIA sensitivity was improved significantly with a combination of these polypeptides. The reactivities of ORF2 polypeptides from genotypes 1 and 4 were similar but that of ORF3 differed with sera from monkeys infected by the two genotypes. Thus, a combination of N- and C-terminal polypeptides of ORF2 from one genotype may be effective in EIAs to detect anti-HEV IgM.

Electron Cryotomography: a New View into Microbial Ultrastructure

Current Opinion in Microbiology. Jun, 2009  |  Pubmed ID: 19427259

Electron cryotomography (ECT) is an emerging technology that allows thin samples such as small bacterial cells to be imaged in 3D in a nearly native state to 'molecular' (approximately 4nm) resolution. As such, ECT is beginning to deliver long-awaited insight into the positions and structures of cytoskeletal filaments, cell wall elements, motility machines, chemoreceptor arrays, internal compartments, and other ultrastructures. Here we briefly explain ECT, review its recent contributions to microbiology, and conclude with a discussion of future prospects.

Mumps Virus Matrix, Fusion, and Nucleocapsid Proteins Cooperate for Efficient Production of Virus-like Particles

Journal of Virology. Jul, 2009  |  Pubmed ID: 19439476

Paramyxovirus particles, like other enveloped virus particles, are formed by budding from membranes of infected cells. To define mumps virus (MuV) proteins important for this process, viral proteins were expressed either singly or in combination in mammalian cells to produce virus-like particles (VLPs). Only the MuV matrix (M) protein when expressed by itself was capable of inducing particle release, but the quantity of these M-alone particles was very small. Efficient production of mumps VLPs occurred only when the M protein was coexpressed together with other viral proteins, with maximum production achieved upon coexpression of the viral M, nucleocapsid (NP), and fusion (F) proteins together. Electron microscopy analysis confirmed that VLPs were morphologically similar to MuV virions. The two MuV glycoproteins were not equal contributors to particle formation. The F protein was a major contributor to VLP production, while the hemagglutinin-neuraminidase protein made a smaller contribution. Evidence for the involvement of class E protein machinery in VLP budding was obtained, with mumps VLP production inhibited upon expression of dominant-negative versions of the class E proteins Vps4A and Chmp4b. Disruption of the sequence 24-FPVI-27 within the MuV M protein led to poor VLP production, consistent with findings of earlier studies of a related sequence, FPIV, important for the budding of parainfluenza virus 5. Together, these results demonstrate that different MuV structural proteins cooperate together for efficient particle production and that particle budding likely involves host class E protein machinery.

The Genome of the Cucumber, Cucumis Sativus L

Nature Genetics. Dec, 2009  |  Pubmed ID: 19881527

Cucumber is an economically important crop as well as a model system for sex determination studies and plant vascular biology. Here we report the draft genome sequence of Cucumis sativus var. sativus L., assembled using a novel combination of traditional Sanger and next-generation Illumina GA sequencing technologies to obtain 72.2-fold genome coverage. The absence of recent whole-genome duplication, along with the presence of few tandem duplications, explains the small number of genes in the cucumber. Our study establishes that five of the cucumber's seven chromosomes arose from fusions of ten ancestral chromosomes after divergence from Cucumis melo. The sequenced cucumber genome affords insight into traits such as its sex expression, disease resistance, biosynthesis of cucurbitacin and 'fresh green' odor. We also identify 686 gene clusters related to phloem function. The cucumber genome provides a valuable resource for developing elite cultivars and for studying the evolution and function of the plant vascular system.

A Non-viral Vector for Potential DMD Gene Therapy Study by Targeting a Minidystrophin-GFP Fusion Gene into the HrDNA Locus

Acta Biochimica Et Biophysica Sinica. Dec, 2009  |  Pubmed ID: 20011980

Gene therapy has emerged as a promising approach for the lethal disorder of Duchenne muscular dystrophy (DMD). Using a novel non-viral delivery system, the human ribosomal DNA (hrDNA) targeting vector, we targeted a minidystrophin-GFP fusion gene into the hrDNA locus of HT1080 cells with a high site-specific integrated efficiency of 10(-5), in which the transgene could express efficiently and continuously. The minidystrophin-GFP fusion protein was easily found to localize on the plasma membrane of HT1080 cells, indicating its possible physiologic performance. Our findings showed that the hrDNA-targeting vector might be highly useful for DMD gene therapy study.

[Raman and FTIR Characteristics of Otolith of Ornamental Carp]

Guang Pu Xue Yu Guang Pu Fen Xi = Guang Pu. Oct, 2009  |  Pubmed ID: 20038039

Otolith is a typical natural biomineral as functional deposit existing in teleost's ears, which is mainly composed of calcium carbonate and organic matter. There is a pair of lapillus, sagitta and asteriscus in fish's inner ear, respectively. The authors compare the asteriscus and lapillus in cultured ornamental carp using FTIR and Raman. The result shows that the mineral phase in lapillus is aragonite, while the mineral phase in asteriscus is vaterite. The pure aragonite and vaterite existing respectively in otolith are of importance as being used as sample to study aragonite/vaterite biomineralization mechanism. Compared with inorganic induced aragonite and vaterite using FTIR and Raman, the authors found that the spectra of aragonite in lapillus are between those of inorganic induced aragonite and other bio induced aragonite; while the spectra of vaterite in asteriscus are similar to those of other bio induced vaterite. It is possible that unstable vaterite was stabilized through the organic effects in biomineralization process.

[Lymph Node Image with Ultrasmall Superparamagnetic Iron Oxide and Comparison with Pathological Result]

Zhongguo Yi Xue Ke Xue Yuan Xue Bao. Acta Academiae Medicinae Sinicae. Apr, 2009  |  Pubmed ID: 19507589

OBJECTIVE: To assess the characteristics of enhanced magnetic resonance image with ultrasmall superparamagnetic iron oxide (USPIO) in the inflammatory and tumor metastatic rabbit model, and explore its relevance with histologic ultrastructural findings. METHODS: Totally 36 New Zealand white rabbits were randomly divided into lymphadenitis group and metastatic group. Complete Freund's adjuvant was injected into the bilateral dorsal footpads of 18 rabbits to set up ipsilateral lymphadenitis model. The other 18 rabbits received a subcutaneous implantation of VX2 tumor cell suspension (1.5 x 10(7) cells/ml) in both thighs to set up metastatic lymph node model. Magnetic resonance scan were performed 24 hours before and after USPIO (90 micromol Fe/kg) injection. T2 values of each lymph node were measured and lymph node T2 enhancement rate was calculated as well. HE staining, Prussian blue staining, and electronic microscopy were performed to observe the pathological microstructure changes and the distribution of the iron particle in lymph node. Relationship between lymph nodes USPIO enhancement and its microstructures were further analyzed. Results Thirty-six lymph nodes in lymphadenitis group showed different degrees of reactive hyperplasia. Twenty-six lymph nodes in metastatic group were invaded by tumor cell. Non-enhanced scan showed mild difference between T2 signal intensity of the two pathological lymph node types. After USPIO enhancement, inflammatory lymph nodes showed distinct T2 signal reduction at the center, and metastatic lymph nodes showed homogenous and faint T2 signal reduction. Enhancement rate of benign and malignant lymph nodes were 57.39% and 29.45% respectively (P < 0.01). HE staining and Prussian blue staining indicated USPIO particles located mainly in the macrophages at inflammatory lymphatic medulla, while paracortical area and cortical area contained relatively much less USPIO particles due to less macrophages distribution. MRI findings were correlated with the pathological results. Electronic microscopy also verified that the majority of USPIO particles were located in the numerous cytophagic bubbles of macrophages. Lymph nodes metastasis including 4 lymph nodes with completed structure destruction due to entire tumor infiltration, 19 lymph nodes with partially lymph node structure destruction but reduced USPIO-contained macrophage numbers or reduced USPIO particles in macrophages, and 3 lymph nodes with only localized foci tumor metastasis at subcapsular area. Conclusions USPIO enhancement pattern of different lymph nodes is closely related to distribution and functional status of the intra-node macrophages. It may affect the accuracy of the lymph node property diagnosis based on USPIO enhanced image.

[Magnetic Resonance Enhancement Features of Inflammatory Lymph Nodes with Ultrasmall Superparamagnetic Iron Oxide in Rabbit Model]

Zhongguo Yi Xue Ke Xue Yuan Xue Bao. Acta Academiae Medicinae Sinicae. Apr, 2009  |  Pubmed ID: 19507597

To study magnetic resonance enhancement features of inflammatory lymph nodes using different doses of ultrasmall superparamagnetic iron oxide (USPIO) particles in order to establish a standardized protocol for USPIO enhanced magnetic resonance imaging of lymph nodes.

[Effect of Penehyclidine Hydrochloride on Oleic Acid-acute Lung Injury in Canine During Ventilation]

Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi = Zhongguo Xiufu Chongjian Waike Zazhi = Chinese Journal of Reparative and Reconstructive Surgery. May, 2009  |  Pubmed ID: 19514584

To observe whether additional penehyclidine hydrochloride (PHC) in mechanical ventilation produces therapeutic effect on oleic acid (OA) induced acute lung injury (ALI) in canine.

A Pharmacogenetic Study of Polymorphisms in Interferon Pathway Genes and Response to Interferon-alpha Treatment in Chronic Hepatitis B Patients

Antiviral Research. Sep, 2009  |  Pubmed ID: 19559055

Certain host genetic polymorphisms in interferon (IFN) signaling pathway genes are reported to be associated with response to IFNalpha therapy. We studied 10 single nucleotide polymorphisms (SNPs) in IFN signaling pathway genes to examine their associations with response to IFN treatment in chronic hepatitis B (CHB) patients. Two hundred and forty-six IFNalpha treatment-naïve CHB patients were enrolled for the present study; all received treatment with IFNalpha alone for 6 months, and the efficacy of the therapy was examined. Ten SNPs in 8 IFN signaling pathway genes were genotyped using a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) protocol. There were no significant differences in allele frequencies and genotype distributions of the 10 SNPs between the response and non-response groups that underwent IFNalpha therapy. However, the frequency of a G-T-G-A 2',5'-oligoadenylate synthetase (OAS) haplotype was significantly higher in the non-response group than that in the response group (16.1% vs. 8.7%, p=0.015). Our study suggested that patients with a G-T-G-A OAS haplotype were less responsive to IFNalpha treatment.

PLK1 Down-regulates Parainfluenza Virus 5 Gene Expression

PLoS Pathogens. Jul, 2009  |  Pubmed ID: 19629176

The paramyxoviruses are a family of negative-sense RNA viruses that includes many important human and animal pathogens. Paramyxovirus RNA synthesis requires the viral phosphoprotein (P) and the large (L) protein. Phosphorylation of P is thought to regulate viral gene expression, though direct proof remains elusive. Recently, we reported that phosphorylation of a specific residue (Ser157) of the P protein of parainfluenza virus 5 (PIV5), a prototypical paramyxovirus, correlates with decreased viral gene expression and cytokine expression in infected cells. Here, we show that: Polo-like kinase 1 (PLK1), a serine/theronine kinase that plays a critical role in regulating the cell cycle, interacts with PIV5 P through the S157 residue; PLK1 inhibition increases viral gene expression; PLK1 over-expression inhibits viral gene expression; and PLK1 directly phosphorylates P in vitro, indicating that PLK1 down-regulates viral gene expression by phosphorylating P. Furthermore, we have determined the PLK1 phosphorylation site on P and found that mutant recombinant PIV5 whose P proteins cannot either bind to or be phosphorylated by PLK1 have similar phenotypes. Increased viral gene expression in PIV5 with mutations in the PLK1 binding/phosphorylation sites correlates with increased induction of cell death and cytokine expression, suggesting that PIV5 limits its viral gene expression to avoid these host effects. It is possible that targeting PLK1 will enhance host innate immune responses, leading to a novel strategy of clearing paramyxovirus infections quickly.

Efficacy of Acupuncture in Prevention of Postoperative Nausea in Cardiac Surgery Patients

The Annals of Thoracic Surgery. Aug, 2009  |  Pubmed ID: 19632409

Coronary artery bypass graft and cardiac valve surgeries are frequently performed in medical facilities in the United States, and postoperative nausea (PON) is a prevalent problem in this patient population. The purpose of this study was to evaluate the efficacy of a single preoperative acupuncture treatment in the prevention of PON in patients undergoing coronary artery bypass graft or cardiac valve surgery, or both.

Results of Mitral Valve Annuloplasty with a Standard-sized Posterior Band: is Measuring Important?

The Journal of Thoracic and Cardiovascular Surgery. Oct, 2009  |  Pubmed ID: 19660356

This study was undertaken to determine hemodynamic and clinical outcomes of annuloplasty with a standard-sized (63 mm) posterior band in adult patients undergoing mitral valve repair for degenerative valve disease.

Expression and Mechanism of Mammalian Target of Rapamycin in Age-related Renal Cell Senescence and Organ Aging

Mechanisms of Ageing and Development. Oct, 2009  |  Pubmed ID: 19698731

The mammalian target of rapamycin (mTOR) is relevant to cell senescence and organismal aging. This study firstly showed that the level of mTOR expression increased with aging in rat kidneys, rat mesangial cells and WI-38 cells (P < 0.05). The levels of phosphorylated-mTOR (p-mTOR), cyclin D1 and p21(WAF1/CIP1/SDI1) expression were significantly higher in WI-38 cells treated with l-leucine (an activator of mTOR) (P < 0.05). The positive staining ratio of senescence-associated beta-galactosidase, number of cells in G1 phase, and cellular volume were all increased in WI-38 cells treated with l-leucine when the cellular population doubling (PD) number was 34, while the above phenotypes did not appear in control group until its PD number reached 40 (P < 0.05). The levels of p-mTOR, cyclin D1, and p21(WAF1/CIP1/SDI1) as well as the aging-related phenotypes were all reduced in cells treated with rapamycin (an inhibitor of mTOR) than in control cells (P < 0.05). These results demonstrated that the level of mTOR was increased in kidney with aging, and that mTOR may promote cellular senescence by regulating the cell cycle through p21(WAF1/CIP1/SDI1), which might provide a new target for preventing renal aging.

Complete Resequencing of 40 Genomes Reveals Domestication Events and Genes in Silkworm (Bombyx)

Science (New York, N.Y.). Oct, 2009  |  Pubmed ID: 19713493

A single-base pair resolution silkworm genetic variation map was constructed from 40 domesticated and wild silkworms, each sequenced to approximately threefold coverage, representing 99.88% of the genome. We identified ~16 million single-nucleotide polymorphisms, many indels, and structural variations. We find that the domesticated silkworms are clearly genetically differentiated from the wild ones, but they have maintained large levels of genetic variability, suggesting a short domestication event involving a large number of individuals. We also identified signals of selection at 354 candidate genes that may have been important during domestication, some of which have enriched expression in the silk gland, midgut, and testis. These data add to our understanding of the domestication processes and may have applications in devising pest control strategies and advancing the use of silkworms as efficient bioreactors.

Radical-molecule Reaction C(3P) + C3H6: Mechanistic Study

The Journal of Physical Chemistry. A. Oct, 2009  |  Pubmed ID: 19731902

The complex triplet potential energy surface for the reaction of ground-state atomic carbon C(3P) with propylene C3H6 is explored at the B3LYP/6-311G(d,p), QCISD/6-311G(d,p), and G3B3 (single-point) levels. Various possible reaction pathways are probed. It is shown that the reaction is initiated by the addition of C(3P) to the C=C bond of C3H6 to generate barrierlessly the three-membered ring isomer 1 CH3-cCHCCH2, followed by the ring-opening process to form 2a trans-CH3CHCCH2, which can easily interconvert to 2b cis-CH3CHCCH2. Starting from 2 (2a, 2b), the most feasible pathway is the internal C-H bond rupture of 2a leading to P4(2CH3CCCH2 + 2H), terminal C-H bond cleavage of 2 (2a,2b) to form P5(2CH3CHCCH + 2H), or direct C-C bond fission of 2b to form P7(2CH2CCH + 2CH3), all of which may have comparable contributions to the title reaction. Much less competitively, 2a takes a 1,2-H-shift to form 5a trans-cis-CH3CHCHCH, followed by a C-C bond rupture leading to P6(1C2H2 + 3CH3CH). Because the intermediates and transition states involved in the feasible pathways all lie below the reactant, the title reaction is expected to be rapid, which is consistent with the measured large rate constant. The present article may provide some useful information for future experimental investigation of the title reaction.

Absence of Pneumocystis Jirovecii Dihydropteroate Synthase Gene Mutations Among Samples from a Group of AIDS Patients in China

Scandinavian Journal of Infectious Diseases. 2009  |  Pubmed ID: 19085210

Pneumocystis jirovecii dihydropteroate synthase (DHPS) gene mutations are associated with failure of sulfur prophylaxis of Pneumocystis pneumonia. Here the P. jirovecii DHPS gene was amplified from bronchoalveolar fluid of 10 AIDS patients in China. No DHPS gene mutations were observed. The results suggest that the prevalence of DHPS mutations in China may be low.

Expression of Enzymes Involved in Synthesis and Metabolism of Estradiol in Human Breast As Studied by Immunocytochemistry and in Situ Hybridization

Histology and Histopathology. Mar, 2009  |  Pubmed ID: 19130396

It is well documented that human breast is actively involved in the local formation of estrogens. To determine the site(s) of action of enzymes involved in synthesis and metabolism of the most potent estrogen estradiol (E2), we have studied the expression of the following enzymes: 3beta-hydroxysteroid dehydrogenase (3-HSD), 17beta-HSD types 1, 2, 5, 7 and 12, aromatase, steroid sulfatase (STS) and estrogen sulfotransferase (EST) 1E1 at the cellular level in breast. Both in situ hybridization and immunocytochemistry were used for enzyme localization in normal breast tissues. For immunocytochemistry, we used rabbit antibodies, while in situ hybridization studies were performed using (35S)-labeled cRNA probes. Similar results were obtained with both approaches. All the enzymes (3beta-HSD; 17beta-HSD types 1, 5, 7 and 12; aromatase) involved in the conversion of circulating dehydroepiandrosterone (DHEA) to E2 as well as STS which converts estradiol sulfate (E2-S) to E2 have been found to be expressed in epithelial cells of acini and/or ducts as well as the stromal cells. Moreover, 17beta-HSD type 2 and EST1E1, two enzymes which inactivate E2, have been also localized in the same cell types. The present results indicate the enzymes which play a role in the synthesis and metabolism of E2 are expressed in both epithelial and stromal cells in human breast.

Complete Pathologic Response After Neoadjuvant Chemoradiotherapy for Esophageal Cancer is Associated with Enhanced Survival

The Annals of Thoracic Surgery. Feb, 2009  |  Pubmed ID: 19161745

Neoadjuvant chemoradiotherapy followed by esophagogastrectomy has become the standard of care for patients with locally advanced esophageal cancer. This report analyzes our experience with this treatment approach.

[Relationship Between the Level of Serum TNF-alpha of Patients with Chronic Hepatitis C and Treatment with Interferon-alpha and the Influencing Factors]

Zhonghua Shi Yan He Lin Chuang Bing Du Xue Za Zhi = Zhonghua Shiyan He Linchuang Bingduxue Zazhi = Chinese Journal of Experimental and Clinical Virology. Apr, 2009  |  Pubmed ID: 20104757

To assess the relationship between the level of serum TNF-alpha of chronic hepatitis C with severity of disease and curative effect of anti-virus therapy with interferon alpha.

Universal Architecture of Bacterial Chemoreceptor Arrays

Proceedings of the National Academy of Sciences of the United States of America. Oct, 2009  |  Pubmed ID: 19805102

Chemoreceptors are key components of the high-performance signal transduction system that controls bacterial chemotaxis. Chemoreceptors are typically localized in a cluster at the cell pole, where interactions among the receptors in the cluster are thought to contribute to the high sensitivity, wide dynamic range, and precise adaptation of the signaling system. Previous structural and genomic studies have produced conflicting models, however, for the arrangement of the chemoreceptors in the clusters. Using whole-cell electron cryo-tomography, here we show that chemoreceptors of different classes and in many different species representing several major bacterial phyla are all arranged into a highly conserved, 12-nm hexagonal array consistent with the proposed "trimer of dimers" organization. The various observed lengths of the receptors confirm current models for the methylation, flexible bundle, signaling, and linker sub-domains in vivo. Our results suggest that the basic mechanism and function of receptor clustering is universal among bacterial species and was thus conserved during evolution.

Association Between Epidermal Growth Factor 61A/G Polymorphism and Hepatocellular Carcinoma Susceptibility in Chinese Patients

Liver International : Official Journal of the International Association for the Study of the Liver. Jan, 2010  |  Pubmed ID: 19840254

Chronic hepatitis B virus (HBV) infection is an important risk factor for hepatocellular carcinoma (HCC) development in China, while little is known of the genetic susceptibility to hepatocarcinogenesis. The epidermal growth factor (EGF) pathway plays an important role in tumorigenesis, including HCC. EGF polymorphisms are associated with susceptibility to several types of cancers. Therefore, this study aimed to assess whether EGF genetic polymorphisms can influence HCC development.

A Nonhuman Primate Model of Alzheimer's Disease Generated by Intracranial Injection of Amyloid-β42 and Thiorphan

Metabolic Brain Disease. Sep, 2010  |  Pubmed ID: 20838863

Alzheimer's disease (AD) is a neurodegenerative disorder characterized by progressive cognitive impairment and neuropathological changes, including the deposition of amyloid-beta (Aβ) peptide. Aged monkeys have proven to be invaluable in the study of AD, as their brains naturally develop amyloid plaques similar to those in AD brains. However, spontaneous development of AD-like pathologies in aged monkeys is time-consuming, often taking several years. Here, we created an experimentally induced AD model in middle-aged (16-17 years) rhesus monkeys by intracranial injection of Aβ42 and thiorphan, an inhibitor of neprilysin that is responsible for Aβ clearance. The working memory capacity of the monkeys in a delayed-response task was little affected following the delivery of Aβ42 and thiorphan. However, the administration of Aβ42 and thiorphan resulted in a significant intracellular accumulation of Aβ in the neurons of the basal ganglia, the cortex, and the hippocampus, accompanied by neuronal atrophy and loss. Moreover, immunohistochemistry revealed a degeneration of choline acetyltransferase-positive cholinergic neurons and an increase of glial fibrillary acidic protein-positive astrocytes. In conclusion, our data demonstrate a primate model of AD generated by combined infusion of Aβ42 and thiorphan, which duplicates a subset of neuropathological changes in AD brains, thereby having implications in the elucidation of this disease.

Effects of Functional Groups and Soluble Matrices in Fish Otolith on Calcium Carbonate Mineralization

Biomedical Materials (Bristol, England). Oct, 2010  |  Pubmed ID: 20844320

Calcium carbonate mineralization is significantly influenced by organic matrices in vivo. The effect mainly relies on functional groups in proteins. In order to study the influence of functional groups on calcium carbonate mineralization, -OH, -NH2 and -COOH groups were grafted onto single crystal silicon chips, and such modified chips were used as substrates in in vitro mineralization experiments. An x-ray photoelectron spectroscopy (XPS) test was conducted to examine the grafting efficiency, and the three groups were successfully grafted. Calcium carbonate mineralization on a modified silicon substrate was examined by a scanning electron microscope (SEM) and x-ray diffraction (XRD), and the results showed that the effects of -OH, -NH2 and -COOH groups were quite different. Furthermore, a water-soluble protein matrix (WSM) and an acid-soluble protein matrix (ASM) extracted from fish otolith were adsorbed onto the -COOH-modified silicon substrate, and the effects of the protein matrices on calcium carbonate mineralization were studied. The results showed that both WSM and ASM of lapillus could mediate aragonite crystallization, but the size and morphology of the formed crystals were different. The WSM and ASM of asteriscus adsorbed on the silicon substrate had little effect on calcium carbonate mineralization; almost all the crystals were calcite, while both asteriscus WSM and ASM in solution could mediate vaterite crystals, and the morphologies of vaterite crystal aggregates were different.

Affinity Purification of an Archaeal DNA Replication Protein Network

MBio. 2010  |  Pubmed ID: 20978540

Nineteen Thermococcus kodakarensis strains have been constructed, each of which synthesizes a different His(6)-tagged protein known or predicted to be a component of the archaeal DNA replication machinery. Using the His(6)-tagged proteins, stable complexes assembled in vivo have been isolated directly from clarified cell lysates and the T. kodakarensis proteins present have been identified by mass spectrometry. Based on the results obtained, a network of interactions among the archaeal replication proteins has been established that confirms previously documented and predicted interactions, provides experimental evidence for previously unrecognized interactions between proteins with known functions and with unknown functions, and establishes a firm experimental foundation for archaeal replication research. The proteins identified and their participation in archaeal DNA replication are discussed and related to their bacterial and eukaryotic counterparts.

Long-term Risk of Aortic Events Following Aortic Valve Replacement in Patients with Bicuspid Aortic Valves

The American Journal of Cardiology. Dec, 2010  |  Pubmed ID: 21094365

Bicuspid aortic valve (BAV) is associated with ascending aortopathy predisposing to aneurysmal dilatation and dissection, even after successful aortic valve replacement (AVR). There is, however, scant evidence on which to make recommendations for prophylactic replacement of the ascending aorta at the time of AVR. The medical records of patients who underwent AVR for BAV without aortic replacement or repair from 1960 to 1995 were reviewed. Follow-up was by review of the medical record and postal questionnaire. Among 1,286 patients, the mean age at operation was 58 ± 14 years. During the follow-up interval (median 12 years, range 0 to 38), there were 13 documented aortic dissections (1%), 11 ascending aortic replacements (0.9%), and 127 documented cases of progressive aortic enlargement (9.9%). Fifteen-year freedom from aortic dissection, enlargement, or replacement was 89% (95% confidence interval [CI] 87% to 91%) and was lower in patients with documented aortic enlargement at the time of AVR (85%, 95% CI 81% to 89%) compared to those whose aortic dimensions were normal (93%, 95% CI 90% to 96%) (p = 0.001). Multivariate predictors of aortic complications included interval (subsequent) AVR (hazard ratio [HR] 3.5, 95% CI 2.3 to 5.4, p <0.001), concomitant coronary artery bypass grafting (HR 2.6, 95% CI 1.7 to 4.0, p <0.001), enlarged aorta (HR 1.8, 95% CI 1.3 to 2.6, p = 0.001), and history of tobacco abuse (HR 1.8, 95% CI 1.2 to 2.6, p = 0.003). Aortic dilatation did not predict mortality. In conclusion, despite a true risk for aortic events after AVR for BAV, the occurrence of aortic dissection was low. Any incremental surgical risk imposed by prophylactic replacement of the ascending aorta must be equally low.

Theoretical Mechanistic Study on the Ion-molecule Reaction of CHCl- with CS2

The Journal of Physical Chemistry. A. Mar, 2010  |  Pubmed ID: 20146458

A detailed theoretical study for the poorly understood ion-molecule reaction of CHCl(-) with CS(2) is explored at the B3LYP/6-311++G(d,p) and CCSD(T)/6-311++G(3df,2p) (single-point) levels. Various possible reaction pathways are considered. On the doublet potential energy surface, five dissociation products are both thermodynamically and kinetically possible. Among these products, P(7) (SCHCl(-) + CS) may be the most favorable product with predominant abundances, whereas P(1) (Cl(-) + SCHCS) and P(2) (Cl(-) + HCCSS) may be the second and third feasible products followed by the almost negligible P(3) (Cl(-) + HSCCS), P(4) (CClS(-) + HCS), and P(6) (S-cCCS(-) + HCl). Because the isomers and transition states involved in the most feasible pathways all lie below the reactant, the title reaction is expected to be fast, which is consistent with the measured large rate constant in recent experiment. The present paper may provide a useful guide for understanding other analogous ion-molecule reactions such as CHF(-) and CHBr(-) with CS(2), COS, and CO(2).

Ancient Human Genome Sequence of an Extinct Palaeo-Eskimo

Nature. Feb, 2010  |  Pubmed ID: 20148029

We report here the genome sequence of an ancient human. Obtained from approximately 4,000-year-old permafrost-preserved hair, the genome represents a male individual from the first known culture to settle in Greenland. Sequenced to an average depth of 20x, we recover 79% of the diploid genome, an amount close to the practical limit of current sequencing technologies. We identify 353,151 high-confidence single-nucleotide polymorphisms (SNPs), of which 6.8% have not been reported previously. We estimate raw read contamination to be no higher than 0.8%. We use functional SNP assessment to assign possible phenotypic characteristics of the individual that belonged to a culture whose location has yielded only trace human remains. We compare the high-confidence SNPs to those of contemporary populations to find the populations most closely related to the individual. This provides evidence for a migration from Siberia into the New World some 5,500 years ago, independent of that giving rise to the modern Native Americans and Inuit.

Immunoglobulin G is Present in a Wide Variety of Soft Tissue Tumors and Correlates Well with Proliferation Markers and Tumor Grades

Cancer. Apr, 2010  |  Pubmed ID: 20186824

The traditional view that immunoglobulin (Ig) is produced only by B lymphocytes has been challenged, because it has been demonstrated that Ig genes and proteins are expressed in epithelial cancer cells. However, whether Ig expression in nonlymphoid cells is limited to epithelial cells is unclear. Because sarcomas differ distinctly from carcinomas in their biologic and clinical features, the authors investigated the question of nonlymphoid IgG expression in soft tissue lesions.

Determinants of Influenza Immunization Uptake in Canadian Youths

Vaccine. Apr, 2010  |  Pubmed ID: 20199757

To describe rate and determinants of influenza vaccination among Canadian youths.

Indications and Outcomes of Surgical Closure of Ventricular Septal Defect in Adults

JACC. Cardiovascular Interventions. Mar, 2010  |  Pubmed ID: 20298987

We sought to review our experience with surgical ventricular septal defect (VSD) closure in adults.

Efficient Fluorescence Resonance Energy Transfer-based Ratiometric Fluorescent Cellular Imaging Probe for Zn(2+) Using a Rhodamine Spirolactam As a Trigger

Analytical Chemistry. Apr, 2010  |  Pubmed ID: 20334436

This letter described the design and synthesis of a novel fluorescein-appended rhodamine spirolactam derivative and its preliminary application as a ratiometric fluorescent cellular imaging probe for Zn(2+). The ratiometric fluorescent signal change of the probe is based on an intramolecular fluorescence resonance energy transfer (FRET) mechanism modulated by a specific metal ion induced ring-opening process of the rhodamine spirolactam (acting as a trigger). In the new developed sensing system, the emission peaks of the two fluorophores are well-resolved, which can avoid the emission spectra overlap problem generally met by spectra-shift type probes and benefits for observation of fluorescence signal change at two different emission wavelengths with high resolution. It also benefits for a large range of emission ratios, thereby a high sensitivity for Zn(2+)detection. Under optimized experimental conditions, the probe exhibits a stable response for Zn(2+) over a concentration range from 2.0 x 10(-7) to 2.0 x 10(-5) M, with a detection limit of 4.0 x 10(-8) M. Most importantly, the novel probe has well solved the problem of serious interferences from other transition metal ions generally met by previously reported typical fluorescent probes for Zn(2+) with the di(2-picolyl)amine moiety as the receptor (in this case, the fluorescence response induced by Cd(2+)is even comparable to that of Zn(2+)) and shows a reversible and fast response toward Zn(2+). All these unique features make it particularly favorable for ratiometric cellular imaging investigations. It has been preliminarily used for ratiometric imaging of Zn(2+) in living cells with satisfying resolution.

Reversible Conduction Block in Isolated Toad Sciatic Nerve by Emulsified Isoflurane

Anesthesia and Analgesia. Apr, 2010  |  Pubmed ID: 20357146

Studies have shown that the local use of volatile anesthetics can produce local anesthetic effects. We designed this study to evaluate the characteristics of nerve conduction block of emulsified isoflurane (EI) and compare its nerve blockade with 1%lidocaine, by measuring compound nerve action potential (CNAP) parameters in isolated toad sciatic nerve.

Repair of the Bicuspid Aortic Valve: a Viable Alternative to Replacement with a Bioprosthesis

The Journal of Thoracic and Cardiovascular Surgery. Jun, 2010  |  Pubmed ID: 20392456

We sought to compare the safety and durability of bicuspid aortic valve repair versus replacement with a bioprosthesis.

A Novel Immunoassay for PreS1 And/or Core-related Antigens for Detection of HBsAg Variants

Journal of Virological Methods. Sep, 2010  |  Pubmed ID: 20451558

A novel immunoassay that detects simultaneously hepatitis B virus (HBV) PreS1 and/or core-related antigens was developed and evaluated for its potential for detecting hepatitis B surface antigen (HBsAg) variants. The detection limits of the assay was 10(2.9+/-0.5)copies/mL (mean+/-SD) for HBsAg-positive sera with different genotypes, and 10(3.5+/-1.2)copies/mL for HBsAg variants sera. The specificity of the assay was 99.9% (95% CI: 99.7-99.9%, 4551 healthy individuals). The sensitivities were 93.9% (95% CI: 92.8-94.9%), 59.3% (95% CI: 38.7-77.6%) and 80% (95% CI: 44.4-97.5%) in three independent groups which include: 2065 hepatitis patients, 27 patients with occult hepatitis B and 10 HBsAg variants, respectively. In addition, a novel premature stop code mutation at position 112 of HBsAg was observed in two patients with chronic hepatitis B with different genotypes.

Possible Pathological Basis for False Diagnoses of Lymph Nodes by USPIO-enhanced MRI in Rabbits

Journal of Magnetic Resonance Imaging : JMRI. Jun, 2010  |  Pubmed ID: 20512896

To determine the cause of misdiagnosis of lymph nodes in ultrasmall superparamagnetic iron oxide (USPIO) enhanced magnetic resonance imaging (MRI) using histological findings in rabbit tumor and inflammatory models.

Electron Cryotomography

Cold Spring Harbor Perspectives in Biology. Jun, 2010  |  Pubmed ID: 20516135

Electron cryotomography (ECT) is an emerging technology that allows thin samples such as macromolecular complexes and small bacterial cells to be imaged in 3-D in a nearly native state to "molecular" ( approximately 4 nm) resolution. As such, ECT is beginning to deliver long-awaited insight into the positions and structures of cytoskeletal fi laments, cell wall elements, motility machines, chemoreceptor arrays, internal compartments, and other ultrastructures. This article describes the technique and summarizes its contributions to bacterial cell biology. For comparable recent reviews, see (Subramaniam 2005; Jensen and Briegel 2007; Murphy and Jensen 2007; Li and Jensen 2009). For reviews on the history, technical details, and broader application of electron tomography in general, see for example (Subramaniam and Milne 2004; Lucić et al. 2005; Leis et al. 2008; Midgley and Dunin-Borkowski 2009).

Möbius Basket Molecule: Structure and Properties

Physical Chemistry Chemical Physics : PCCP. Aug, 2010  |  Pubmed ID: 20532360

Expanding the non-knot region of the famous Möbius strip with topological one-sided characteristics, an interesting structure of a Möbius basket molecule with all real frequencies was obtained at the B3LYP/6-31G(d) level for the first time. This structure contains six fused five-membered pyrrole rings contrasting with the isolated pentagon rule for the fullerenes. The twisted handle joints the outer and inner surfaces of the bowl to form a one-sided container molecule. Comparing the Möbius basket to its isomers of a Möbius strip and a normal basket, the framework shape effects on the structure and properties are systematically exhibited. Especially, (1) the basket-making effect increases kinetic stability (the HOMO-LUMO gap increases from 1.116 eV for Möbius strip to 1.608 eV for Möbius basket); (2) from the normal basket to the Möbius basket, the twisting effect obviously increases the static first hyperpolarizability (from 2836 to 3773 au) and IP (from 6.622 to 6.857 eV). It is found that the aza atom, knot, the bowl, and the combination of the knot and bowl units are important regulating factors for the charge transfer (CT) direction in the crucial transitions. This provides the possibility to control the charge transfer direction in crucial transitions by variation of the structures, which is important for the designs of the new optical and photoelectric materials and devices with good performances.

Phosphorylation and Dephosphorylation Among Dif Chemosensory Proteins Essential for Exopolysaccharide Regulation in Myxococcus Xanthus

Journal of Bacteriology. Sep, 2010  |  Pubmed ID: 20543066

Myxococcus xanthus social gliding motility, which is powered by type IV pili, requires the presence of exopolysaccharides (EPS) on the cell surface. The Dif chemosensory system is essential for the regulation of EPS production. It was demonstrated previously that DifA (methyl-accepting chemotaxis protein [MCP]-like), DifC (CheW-like), and DifE (CheA-like) stimulate whereas DifD (CheY-like) and DifG (CheC-like) inhibit EPS production. DifD was found not to function downstream of DifE in EPS regulation, as a difD difE double mutant phenocopied the difE single mutant. It has been proposed that DifA, DifC, and DifE form a ternary signaling complex that positively regulates EPS production through the kinase activity of DifE. DifD was proposed as a phosphate sink of phosphorylated DifE (DifE approximately P), while DifG would augment the function of DifD as a phosphatase of phosphorylated DifD (DifD approximately P). Here we report in vitro phosphorylation studies with all the Dif chemosensory proteins that were expressed and purified from Escherichia coli. DifE was demonstrated to be an autokinase. Consistent with the formation of a DifA-DifC-DifE complex, DifA and DifC together, but not individually, were found to influence DifE autophosphorylation. DifD, which did not inhibit DifE autophosphorylation directly, was found to accept phosphate from autophosphorylated DifE. While DifD approximately P has an unusually long half-life for dephosphorylation in vitro, DifG efficiently dephosphorylated DifD approximately P as a phosphatase. These results support a model where DifE complexes with DifA and DifC to regulate EPS production through phosphorylation of a downstream target, while DifD and DifG function synergistically to divert phosphates away from DifE approximately P.

Sequencing of 50 Human Exomes Reveals Adaptation to High Altitude

Science (New York, N.Y.). Jul, 2010  |  Pubmed ID: 20595611

Residents of the Tibetan Plateau show heritable adaptations to extreme altitude. We sequenced 50 exomes of ethnic Tibetans, encompassing coding sequences of 92% of human genes, with an average coverage of 18x per individual. Genes showing population-specific allele frequency changes, which represent strong candidates for altitude adaptation, were identified. The strongest signal of natural selection came from endothelial Per-Arnt-Sim (PAS) domain protein 1 (EPAS1), a transcription factor involved in response to hypoxia. One single-nucleotide polymorphism (SNP) at EPAS1 shows a 78% frequency difference between Tibetan and Han samples, representing the fastest allele frequency change observed at any human gene to date. This SNP's association with erythrocyte abundance supports the role of EPAS1 in adaptation to hypoxia. Thus, a population genomic survey has revealed a functionally important locus in genetic adaptation to high altitude.

Validation of Clinical Scores Predicting Severe Acute Kidney Injury After Cardiac Surgery

American Journal of Kidney Diseases : the Official Journal of the National Kidney Foundation. Oct, 2010  |  Pubmed ID: 20630639

Acute kidney injury (AKI) requiring renal replacement therapy (RRT) in patients undergoing cardiac surgery is associated strongly with adverse patient outcomes. Recently, 3 predictive risk models for RRT have been developed. The aims of our study are to validate the predictive scoring models for patients requiring postoperative RRT and test applicability to the broader spectrum of patients with postoperative severe AKI.

Association of NKG2D Genetic Polymorphism with Susceptibility to Chronic Hepatitis B in a Han Chinese Population

Journal of Medical Virology. Sep, 2010  |  Pubmed ID: 20648603

Natural killer (NK) cells are important antiviral effectors of innate immunity because of their contribution to virus elimination. NK cell-mediated immunological reaction to hepatitis B virus (HBV) infection depends on a fine balance between inhibitory and activating receptors. The aim of the study was to investigate genetic polymorphisms in NK cell receptors (NKR)-KLRD1 (CD94), KLRK1 (NKG2D), KLRC4 (NKG2F), and KLRC1 (NKG2A)-to evaluate the association of NKR genetic polymorphisms with susceptibility to chronic hepatitis B in a Han Chinese population. Twelve single nucleotide polymorphisms (SNPs), including rs2302489 in CD94; rs2255336, rs2617160, rs7980470, rs 2734565, and rs17513986 in NKG2D; rs2617170, rs17549004, and rs3825295 in NKG2F; rs2734414, rs7301582, and rs2734440 in NKG2A, were selected in the present study. SNP genotyping was undertaken in 500 Han Chinese patients (285 patients with chronic hepatitis B and 215 patients who cleared HBV spontaneously) by a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and by the TaqMan method. Single marker association analysis was conducted and the SNP rs2617160 with a TT genotype in NKG2D was associated significantly with an increased risk of chronic hepatitis B (P = 0.044; OR = 1.49; 95% CI = 1.01-2.19). Haplotype analysis with multiple loci indicated that there was no significant association between the haplotypes of the NKR genes and susceptibility to chronic hepatitis B. The SNP rs2617160 in NKG2D associated with susceptibility to chronic hepatitis B in a Han Chinese population.

Vascular Endothelial Growth Factor-C Associated with Computed Tomography Used in the Diagnosis of Lymph Node Metastasis of Bladder Carcinoma

Archives of Medical Research. Nov, 2010  |  Pubmed ID: 21199729

Vascular endothelial growth factor C (VEGF-C) has been demonstrated to stimulate the growth of lymphatic vascular endothelium. The purpose of this study is to determine whether VEGF-C associated with computed tomography (CT) has a relationship with lymph node metastasis in bladder transitional cell carcinoma (BTCC).

Environmentally Benign Methanolysis of Polycarbonate to Recover Bisphenol A and Dimethyl Carbonate in Ionic Liquids

Journal of Hazardous Materials. Feb, 2010  |  Pubmed ID: 19773125

An environmentally friendly strategy for methanolysis of polycarbonate (PC) to recover bisphenol A (BPA) and dimethyl carbonate (DMC) was developed in which PC could be methanolyzed in an ionic liquid without any acid or base catalyst under moderate conditions. The effects of ionic liquid kinds, temperature, time, and methanol dosage on methanolysis results of PC were examined. It was showed that the methanolysis conversion of PC was almost 100% and the yields of both BPA and DMC were over 95% in presence of ionic liquid 1-n-butyl-3-methylimidazolium chloride ([Bmim][Cl]) and under the conditions of m(PC):m(CH(3)OH):m([Bmim][Cl])=2:3:2, reaction temperature 105 degrees C and time 2.5h. After easily separated from the product, the ionic liquid could be reused 8 times without obvious decrease in the conversion of PC and yields of BPA and DMC.

Effect of Unassembled Phage Protein Complexes on the Attachment to Cellulose of Genetically Modified Bacteriophages Containing Cellulose Binding Modules

Colloids and Surfaces. B, Biointerfaces. Apr, 2010  |  Pubmed ID: 20071155

One way to make bioactive paper is to incorporate in it bacteriophages, which can capture and deactivate bacteria. The release of newly assembled phages from lysed bacteria can also serve as a measure of the number of bacteria that were deactivated. We have studied the deposition of genetically modified phages, containing cellulose binding modules, on model cellulose surfaces by evanescent wave light scattering. It was shown that unassembled protein complexes, released with the phages during the lysis of bacteria, limit the deposition of phages to about one per 10-20 microm2, as there are far more unassembled protein complexes than phages. The implication is that a few thousand phages can attach to a cellulose fiber, with most of the surface area taken up by protein complexes. This is more than sufficient for the production of bioactive paper. The unassembled protein complexes may be beneficial, as they will prevent non-specific deposition of bacteria. Thus, it may not be necessary to separate the protein complexes from phages in an expensive purification operation.

Pericardial Effusion After Cardiac Surgery: Risk Factors, Patient Profiles, and Contemporary Management

The Annals of Thoracic Surgery. Jan, 2010  |  Pubmed ID: 20103217

We aimed to review recent experience at our institution in the diagnosis and treatment of pericardial effusion after cardiac surgery and to identify risk factors for its development.

Biohydrogen Production from Dairy Manures with Acidification Pretreatment by Anaerobic Fermentation

Environmental Science and Pollution Research International. Feb, 2010  |  Pubmed ID: 19499259

Hydrogen is a clean and efficient energy source and has been deemed as one of the most promising carriers of new energy for the future. From an engineering point of view, producing hydrogen by mixed cultures is generally preferred because of lower cost, ease of control, and the possible use of organic waste as feedstock. The biological hydrogen production has been intensively studied in recent decades. So far, most investigates of biohydrogen production are still confined to using pure carbohydrates and carbohydrate-rich wastewater. Nowadays, the large amounts of livestock manure, which come from cattle feedlots, poultry, and swine buildings, are causing a major environmental issue because it has become a primary source of odors, gases, dust, and groundwater contamination. The increasingly stringent requirements for pollution control on livestock manures are challenging the scientific community to develop new waste treatment strategies. Thus, there is a pressing need to develop nonpolluting and renewable energy source utilizing the organic waste (e.g., livestock manure). It is well known that anaerobic digestion had successfully been used for the disposal of manures to produce methane in the last two decades. Recently, an alternative strategy has been developed to convert livestock manures (e.g., dairy manures) to biohydrogen as a high value-added clean energy source instead of methane. However, little information is available on hydrogen production from dairy manure via the mixed anaerobic microbe. As far as we know, the hydrogen production is habitually accompanied with production of volatile fatty acids (VFAs), such as acetate, butyrate, and propionate, which are also an optimal feedstock for production of methane by anaerobic digestion. Provided that the biohydrogen production from dairy manure is further combined with the anaerobic digestion of the effluent from the producing hydrogen reactor that would be a one-stone two-bird paradigm, it not only produces a clean and readily usable biologic energy but also cleans up simultaneously the environment in a sustainable fashion.

Clinical Accuracy of RIFLE and Acute Kidney Injury Network (AKIN) Criteria for Acute Kidney Injury in Patients Undergoing Cardiac Surgery

Critical Care (London, England). 2011  |  Pubmed ID: 21232094

The RIFLE (risk, injury, failure, loss of kidney function, and end-stage renal failure) classification for acute kidney injury (AKI) was recently modified by the Acute Kidney Injury Network (AKIN). The two definition systems differ in several aspects, and it is not clearly determined which has the better clinical accuracy.

Human Leukocyte Antigen Class I and Class II Genes Polymorphisms Might Be Associated with Interferon α Therapy Efficiency of Chronic Hepatitis B

Antiviral Research. Mar, 2011  |  Pubmed ID: 21241740

Certain host genetic polymorphisms in human leukocyte antigen (HLA) genes are reported to be associated with response to interferon α (IFNα) therapy. Two hundred and eighteen IFNα treatment-naïve chronic hepatitis B (CHB) patients were enrolled in the present study. HLA-A, B, C and DQA1, DQB1, DRB1 gene alleles were detected by polymerase chain reaction-sequencing based typing (PCR-SBT) and PCR-sequence specific primer (PCR-SSP), respectively. Frequencies of HLA-DQB1*0303 and DRB1*08 in response group were clearly lower than those in nonresponse group (P=0.019, OR=1.81, 95%CI=1.07-3.15; P=0.031, OR=2.43, 95%CI=1.02-5.98, respectively). Frequencies of haplotype *1101-*4601-*0102 (HLA-A, B, C) and haplotype *0302-*0303-*09 (HLA-DQA1, DQB1, DRB1) were clearly lower than those in nonresponse group (P=0.009, OR=4.84, 95%CI=1.29-19.48; P=0.031, OR=1.94, 95%CI=1.01-3.73, respectively). These results suggest that patients with HLA-DQB1*0303 or DRB1*08 alleles, and haplotype *1101-*4601-*0102 (HLA-A, B, C) or haplotype *0302-*0303-*09 (HLA-DQA1, DQB1, DRB1), might be less responsive to IFNα treatment.

Reducing Pulmonary Injury by Hyperbaric Oxygen Preconditioning During Simulated High Altitude Exposure in Rats

The Journal of Trauma. Sep, 2011  |  Pubmed ID: 21248653

Hyperbaric oxygen preconditioning (HBO₂P + HAE) has been found to be beneficial in preventing the occurrence of ischemic damage to brain, spinal cord, heart, and liver in several disease models. In addition, pulmonary inflammation and edema are associated with a marked reduction in the expression levels of both aquaporin (AQP) 1 and AQP5 in the lung. Here, the aims of this study are first to ascertain whether acute lung injury can be induced by simulated high altitude in rats and second to assess whether HBO2P + HAE is able to prevent the occurrence of the proposed high altitude-induced ALI.

Functional Tricuspid Regurgitation at the Time of Mitral Valve Repair for Degenerative Leaflet Prolapse: the Case for a Selective Approach

The Journal of Thoracic and Cardiovascular Surgery. Sep, 2011  |  Pubmed ID: 21277597

It is not clear whether clinically silent tricuspid valve regurgitation should be addressed at the time of mitral valve repair for severe mitral regurgitation due to leaflet prolapse. We examined the clinical and echocardiographic outcomes of patients with tricuspid regurgitation who underwent only mitral valve repair.

[Protective Effect of Compound Tianpupian (TPP) Against H(2)O(2)-induced Apoptosis of Murine Splenic Lymphocytes]

Zhongguo Shi Yan Xue Ye Xue Za Zhi / Zhongguo Bing Li Sheng Li Xue Hui = Journal of Experimental Hematology / Chinese Association of Pathophysiology. Feb, 2011  |  Pubmed ID: 21362254

The aim of this study was to explore the protective effect of compound tianpupian (TPP) against (2)O(2)-induced the apoptosis of murine splenic lymphocytes and its mechanism. The cell apoptosis rate was detected by MTT method; the cell apoptosis and mitochondrial membrance potential were detected by flow cytometry (FCM) with Annexi-V/PI double staining and JC-1 staining method, respectively; and caspase 3 relative activity was determined by colorimetry. The results indicated that after treating with (2)O(2), the absorbance value of cultured lymphocytes and the red/green ratio of JC-1 were reduced, and the apoptotic rate and caspase 3 activity were increased, coculture of (2)O(2)-treated cells with compound TPP increased the cell absorbance ratio and red/green rate of JC-1, while reduced the apoptosis rate and caspase 3 activity. It is concluded that compound TPP alleviates intracellular oxidative damages and dose-dependently inhibited apoptosis of murine splenic lymphocytes through reducing mitochondrial membrane potential and inhibiting caspase 3 activity. This suggests that compound TPP is a potential anti-apoptotic agent.

Retroperitoneal Laparoscopic Dismembered Pyeloplasty with a Novel Technique of JJ Stenting in Children

BJU International. Sep, 2011  |  Pubmed ID: 21401848

• To report our experience with retroperitoneal laparoscopic dismembered pyeloplasty for pelvi-ureteric junction (PUJ) obstruction in children.

Evaluation of Genetic Susceptibility Loci for Chronic Hepatitis B in Chinese: Two Independent Case-control Studies

PloS One. 2011  |  Pubmed ID: 21408128

A recent genome-wide scan has identified two genetic variants in the HLA-DP region strongly associated with hepatitis B infection in Japanese. This study evaluates the effects of these risk variants in Chinese, where the HBV infection is the most popular in the world.

Long Helical Filaments Are Not Seen Encircling Cells in Electron Cryotomograms of Rod-shaped Bacteria

Biochemical and Biophysical Research Communications. Apr, 2011  |  Pubmed ID: 21419100

How rod-shaped bacteria form and maintain their shape is an important question in bacterial cell biology. Results from fluorescent light microscopy have led many to believe that the actin homolog MreB and a number of other proteins form long helical filaments along the inner membrane of the cell. Here we show using electron cryotomography of six different rod-shaped bacterial species, at macromolecular resolution, that no long (> 80 nm) helical filaments exist near or along either surface of the inner membrane. We also use correlated cryo-fluorescent light microscopy (cryo-fLM) and electron cryo-tomography (ECT) to identify cytoplasmic bundles of MreB, showing that MreB filaments are detectable by ECT. In light of these results, the structure and function of MreB must be reconsidered: instead of acting as a large, rigid scaffold that localizes cell-wall synthetic machinery, moving MreB complexes may apply tension to growing peptidoglycan strands to ensure their orderly, linear insertion.

The Impact of Competitive Flow on Distal Coronary Flow and on Graft Flow During Coronary Artery Bypass Surgery

Interactive Cardiovascular and Thoracic Surgery. Jun, 2011  |  Pubmed ID: 21441253

To determine the impact of left anterior descending-competitive flow (LAD-CF) on distal coronary flow (LAD-DF) and on left internal mammary artery-graft flow (LIMA-GF), we performed a quantitative blood-flow analysis in a swine model of a LIMA-to-LAD coronary artery bypass graft (CABG). In six swine, a LIMA-to-LAD CABG was performed. LAD blood-flow was measured bilaterally to the LIMA-to-LAD anastomosis, in the LIMA and in the pulmonary artery (cardiac output, CO) along with the LIMA pulsatility index (LIMA-PI) and the left ventricular pressure (LVP). PreCABG measurements were followed by postCABG measurements at five levels of LAD-CF: 100%, 75%, 50%, 25% and 0% after gradually snaring down a snare placed proximally of the LAD-CF flow-probe. PreCABG CO and LVP remained unchanged postCABG. LAD-DF was reduced significantly postCABG (-33%, P<0.0001). Reduction of the LAD-CF (at 75%, 50%, 25% and 0%) resulted in significant increase of LIMA-GF (+38%, +63%, +113%, +225%, P<0.036 at all LAD-CF levels), reduced PI (6.8, 5.7, 4.1, 3.1, 2.5) with simultaneous increase of LAD-DF (+8%, P=NS, +8%, P=NS, +17%, P=NS, +50%, P=0.0044). Decreased LAD-CF resulted in increased LAD-DF, increased LIMA-GF and decreased LIMA-PI. To the best of our knowledge, this is the first study where blood-flow was directly and simultaneously measured in all the components of the LIMA-to-LAD anastomosis.

A Novel DNA Nuclease is Stimulated by Association with the GINS Complex

Nucleic Acids Research. Aug, 2011  |  Pubmed ID: 21459845

Chromosomal DNA replication requires the spatial and temporal coordination of the activities of several complexes that constitute the replisome. A previously uncharacterized protein, encoded by TK1252 in the archaeon Thermococcus kodakaraensis, was shown to stably interact with the archaeal GINS complex in vivo, a central component of the archaeal replisome. Here, we document that this protein (TK1252p) is a processive, single-strand DNA-specific exonuclease that degrades DNA in the 5' → 3' direction. TK1252p binds specifically to the GINS15 subunit of T. kodakaraensis GINS complex and this interaction stimulates the exonuclease activity in vitro. This novel archaeal nuclease, designated GINS-associated nuclease (GAN), also forms a complex in vivo with the euryarchaeal-specific DNA polymerase D. Roles for GAN in replisome assembly and DNA replication are discussed.

Spinal Cord Protection During Open Repair of Thoracic and Thoracoabdominal Aortic Aneurysms Using Profound Hypothermia and Circulatory Arrest

Journal of the American College of Surgeons. Apr, 2011  |  Pubmed ID: 21463811

Reduced risk of paraplegia is argued as an advantage of endovascular repair of descending thoracic aortic aneurysms (DTA) and thoracoabdominal aortic aneurysms (TAAA); however, paraplegia rates with open repair vary widely.

Synthesis and Bioactivity of New Finasteride Conjugate

Bioorganic & Medicinal Chemistry Letters. Jun, 2011  |  Pubmed ID: 21515045

Finasteride is a synthetic 4-azasteroid compound that acts by inhibiting type II 5α-reductase, the enzyme that converts the androgen testosterone to 5α-dihydrotestosterone. It was approved by the US FDA for the treatment of benign prostatic hyperplasia and male pattern baldness. Here the acylation product of Finasteride C-18 amide N-polimod was synthesized by employing acylation reaction with polimod amide as a pivotal intermediate. The structure of the key intermediate and target molecule was confirmed by infrared spectrum, (1)H NMR and (13)C NMR spectra and mass spectrum, and the inhibition of the steroid 5α-reductase and the rats' benign prostatic hyperplasia by the new Finasteride conjugate and Finasteride was also determined. The inhibition of the Finasteride conjugate on 5α-reductase was stronger than that of Finasteride. Prostate hyperplasia of rats was reduced by Finasteride conjugate treatment similar to the Finasteride treatment. However, the Finasteride conjugate treated animals showed better viable condition than the Finasteride treated ones, suggesting the new compound may have improved toxicity profile than Finasteride.

Congenitally Corrected Transposition of the Great Arteries Ventricular Function at the Time of Systemic Atrioventricular Valve Replacement Predicts Long-term Ventricular Function

Journal of the American College of Cardiology. May, 2011  |  Pubmed ID: 21565637

The objective was to evaluate the systemic ventricular ejection fraction (SVEF) at the time of systemic atrioventricular valve (SAVV) replacement as a predictor of SVEF ≥1 year after surgery in patients with congenitally corrected transposition of the great arteries (CCTGA).

Hypobaric Hypoxia Preconditioning Attenuates Acute Lung Injury During High-altitude Exposure in Rats Via Up-regulating Heat-shock Protein 70

Clinical Science (London, England : 1979). Sep, 2011  |  Pubmed ID: 21599636

HHP (hypobaric hypoxia preconditioning) induces the overexpression of HSP70 (heat-shock protein 70), as well as tolerance to cerebral ischaemia. In the present study, we hypothesized that HHP would protect against HAE (high-altitude exposure)-induced acute lung injury and oedema via promoting the expression of HSP70 in lungs prior to the onset of HAE. At 2 weeks after the start of HHP, animals were exposed to a simulated HAE of 6000 m in a hypobaric chamber for 24 h. Immediately after being returned to ambient pressure, the non-HHP animals had higher scores of alveolar oedema, neutrophil infiltration and haemorrhage, acute pleurisy (e.g. increased exudate volume, increased numbers of polymorphonuclear cells and increased lung myeloperoxidase activity), increased pro-inflammatory cytokines [e.g. TNF-α (tumour necrosis factor-α), IL (interleukin)-1β and IL-6], and increased cellular ischaemia (i.e. glutamate and lactate/pyruvate ratio) and oxidative damage [glycerol, NOx (combined nitrate+nitrite) and 2,3-dihydroxybenzoic acid] markers in the BALF (bronchoalveolar fluid). HHP, in addition to inducing overexpression of HSP70 in the lungs, significantly attenuated HAE-induced pulmonary oedema, inflammation, and ischaemic and oxidative damage in the lungs. The beneficial effects of HHP in preventing the occurrence of HAE-induced pulmonary oedema, inflammation, and ischaemic and oxidative damage was reduced significantly by pretreatment with a neutralizing anti-HSP70 antibody. In conclusion, HHP may attenuate the occurrence of pulmonary oedema, inflammation, and ischaemic and oxidative damage caused by HAE in part via up-regulating HSP70 in the lungs.

Novel Method for Double-J Stenting in Retroperitoneal Laparoscopic Dismembered Pyeloplasty

Urology. Feb, 2011  |  Pubmed ID: 20708779

To describe a novel technique of double-J stenting in laparoscopic pyeloplasty.

A Model for the Preferential Delivery of Isoflurane to the Spinal Cord of the Goat

Veterinary Journal (London, England : 1997). Feb, 2011  |  Pubmed ID: 20739204

To identify the blood supply of the caprine central nervous system, six anaesthetised goats were perfused with coloured suspension into the brachiocephalic artery, the aorta, the iliac artery and the femoral artery. The subsequent distribution indicated that the brain and the main segments of the spinal cord were supplied by the brachiocephalic artery and aorta, respectively. Ten similarly anaesthetised goats then received emulsified isoflurane randomly via either the proximal part of the descending aorta (arterial group) or an ear vein (venous group). In the arterial group, the isoflurane partial pressure (P(iso)) in femoral arterial blood was almost double the P(iso) in jugular venous blood. The model showed that preferential delivery of isoflurane to the goat spinal cord in situ was possible and could be used for further research into the mechanisms of anaesthetic action, particularly factors affecting immobility.

Relationship Between Left Ventricular Mass, Wall Thickness, and Survival After Subaortic Septal Myectomy for Hypertrophic Obstructive Cardiomyopathy

The Journal of Thoracic and Cardiovascular Surgery. Feb, 2011  |  Pubmed ID: 20832083

In natural history studies, increased left ventricular mass and wall thickness are strongly associated with increased late mortality in patients with hypertrophic cardiomyopathy. Our objective was to determine the impact of left ventricular mass and wall thickness on survival after myectomy for hypertrophic obstructive cardiomyopathy.

Can the Impact of Change of Surgical Teams in Cardiovascular Surgery Be Measured by Operative Mortality or Morbidity? A Propensity Adjusted Cohort Comparison

Annals of Surgery. Feb, 2011  |  Pubmed ID: 21173693

Our objective was to examine the impact of team changeover and unfamiliar teams in cardiovascular surgery on traditional clinical outcome measures.

Function of the Small Hydrophobic Protein of J Paramyxovirus

Journal of Virology. Jan, 2011  |  Pubmed ID: 20980504

At 18,954 nucleotides, the J paramyxovirus (JPV) genome is one of the largest in the family Paramyxoviridae, consisting of eight genes in the order 3'-N-P/V/C-M-F-SH-TM-G-L-5'. To study the function of novel paramyxovirus genes in JPV, a plasmid containing a full-length cDNA clone of the genome of JPV was constructed. In this study, the function of the small hydrophobic (SH) protein of JPV was examined by generating a recombinant JPV lacking the coding sequence of the SH protein (rJPVΔSH). rJPVΔSH was viable and had no growth defect in tissue culture cells. However, more tumor necrosis factor alpha (TNF-α) was produced during rJPVΔSH infection, suggesting that SH plays a role in inhibiting TNF-α production. rJPVΔSH induced more apoptosis in tissue culture cells than rJPV. Virus-induced apoptosis was inhibited by neutralizing antibody against TNF-α, suggesting that TNF-α contributes to JPV-induced apoptosis in vitro. The expression of JPV SH protein inhibited TNF-α-induced NF-κB activation in a reporter gene assay, suggesting that JPV SH protein can inhibit TNF-α signaling in vitro. Furthermore, infection of mice with rJPVΔSH induced more TNF-α expression, indicating that SH plays a role in blocking TNF-α expression in vivo.

Regression in Left Ventricular Mass After Aortic Valve Replacement for Chronic Aortic Regurgitation is Unrelated to Prosthetic Valve Size

The Journal of Thoracic and Cardiovascular Surgery. Aug, 2011  |  Pubmed ID: 21074186

We examined the role of prosthesis-patient mismatch on left ventricular mass regression after aortic valve replacement for chronic aortic valve regurgitation.

Emulsified Isoflurane Preconditioning Protects Against Liver and Lung Injury in Rat Model of Hemorrhagic Shock

The Journal of Surgical Research. Dec, 2011  |  Pubmed ID: 20850773

Isoflurane has demonstrated protective effects against ischemia/reperfusion injury in some organs. In this study, using the hemorrhagic shock model, we investigated whether emulsified isoflurane preconditioning protected against liver and lung injury caused by massive surgical blood loss.

Structural Diversity of Bacterial Flagellar Motors

The EMBO Journal. Jul, 2011  |  Pubmed ID: 21673657

The bacterial flagellum is one of nature's most amazing and well-studied nanomachines. Its cell-wall-anchored motor uses chemical energy to rotate a microns-long filament and propel the bacterium towards nutrients and away from toxins. While much is known about flagellar motors from certain model organisms, their diversity across the bacterial kingdom is less well characterized, allowing the occasional misrepresentation of the motor as an invariant, ideal machine. Here, we present an electron cryotomographical survey of flagellar motor architectures throughout the Bacteria. While a conserved structural core was observed in all 11 bacteria imaged, surprisingly novel and divergent structures as well as different symmetries were observed surrounding the core. Correlating the motor structures with the presence and absence of particular motor genes in each organism suggested the locations of five proteins involved in the export apparatus including FliI, whose position below the C-ring was confirmed by imaging a deletion strain. The combination of conserved and specially-adapted structures seen here sheds light on how this complex protein nanomachine has evolved to meet the needs of different species.

Rescue of Wild-type Mumps Virus from a Strain Associated with Recent Outbreaks Helps to Define the Role of the SH ORF in the Pathogenesis of Mumps Virus

Virology. Aug, 2011  |  Pubmed ID: 21676427

Mumps virus (MuV) causes acute infections in humans. In recent years, MuV has caused epidemics among highly vaccinated populations. The largest outbreak in the U.S. in the past 20 years occurred in 2005-2006 with over 5000 reported cases in which the majority of the cases was in vaccinated young adults. We sequenced the complete genome of a representative strain from the epidemic (MuV-IA). MuV-IA is a member of genotype G, the same genotype of MuV that was associated with the outbreak in the UK in 2004-2005. We constructed a reverse genetics system for MuV-IA (rMuV-IA), and rescued a virus lacking the open reading frame (ORF) of the SH gene (rMuV∆SH). rMuV∆SH infection in L929 cells induced increased NF-κB activation, TNF-α production and apoptosis compared to rMuV-IA. rMuV∆SH was attenuated in an animal model. These results indicated that the SH ORF of MuV plays a significant role in interfering with TNF-α signaling and viral pathogenesis during virus infection.

Targeting the Kynurenine Pathway As a Potential Strategy to Prevent and Treat Alzheimer's Disease

Medical Hypotheses. Sep, 2011  |  Pubmed ID: 21683531

Alzheimer's disease (AD) is a progressive neurodegenerative disorder of the elderly accounting for the vast majority of dementia. Recently, many studies have implicated the role of inflammatory response, especially neuroinflammatory response in the development and progression of AD. However, the underlying mechanism of how inflammatory response induces AD is unknown. Kynurenine pathway is a major route of the amino acid tryptophan catabolism, resulting in the production of nicotine adenine dinucleotide and other neuroactive intermediates: quinolinic acid (QA) and kynurenic acid (KA). QA exerts different toxic effects, including over-activation of N-methyl-d-aspartate (NMDA) receptor and excitotoxicity, synaptic dysfunction and neuronal death. On the other hand, KA is identified as the only endogenous NMDA receptor antagonist and could modulate neurotoxic effects of QA. We hypothesize that an activated kynurenine pathway induced by inflammatory cytokines would generate more neurotoxic metabolites, which could be closely related to the pathogenesis of AD in elderly patients. Moreover, some measures, which facilitate KA synthesis and reduce the formation of QA, may emerge as a new therapeutic strategy against AD.

NAD Blocks High Glucose Induced Mesangial Hypertrophy Via Activation of the Sirtuins-AMPK-mTOR Pathway

Cellular Physiology and Biochemistry : International Journal of Experimental Cellular Physiology, Biochemistry, and Pharmacology. 2011  |  Pubmed ID: 21691086

Since the discovery of NAD-dependent deacetylases, Sirtuins, it has been recognized that maintaining intracellular levels of NAD is crucial for the management of stress-response of cells. Here we show that high glucose(HG)-induced mesangial hypertrophy is associated with loss of intracellular levels of NAD. This study was designed to investigate the effect of NAD on HG-induced mesangial hypertrophy.

Risk Factors and Early Outcomes of Multiple Reoperations in Adults with Congenital Heart Disease

The Annals of Thoracic Surgery. Jul, 2011  |  Pubmed ID: 21718837

Advances in treatment of congenital heart disease (CHD) have resulted in most patients surviving to adulthood. Despite surgical "correction," the need for reoperation(s) persists, and there are few outcome data. This study examined early postoperative results to determine risk factors for cardiac injury and early death in adults with CHD undergoing repeat median sternotomy.

Oculocutaneous Albinism Type 3 (OCA3): Analysis of Two Novel Mutations in TYRP1 Gene in Two Chinese Patients

Cell Biochemistry and Biophysics. Dec, 2011  |  Pubmed ID: 21739261

Oculocutaneous albinism (OCA) is a genetic disease characterized by the reduction or deficiency of melanin in eyes, skin, and hair. OCA exhibits genetic heterogeneity. Presently, there are four types of OCA named as OCA1, OCA2, OCA3, and OCA4. OCA3 is more common in African born blacks but rarely found in other ethnic populations. Our recent genotyping of patients with OCA of Chinese descent has identified two patients who were not OCA1, OCA2, or OCA4. Examination and analysis of the TYRP1 gene identified them to be having OCA3. PCR and DNA sequencing analysis found that the mutant TYPR1 alleles were present in each of the two patients, c.780-791del/c.1067G>A (p.R356Q) and c.625G>TT (p.G209LfsX1)/c.643C>T (p.H215Y). The c.780-791del and c.1067G>A mutations have been already reported. However, the c.625G>TT and c.643C>T mutations have not been previously reported and were found to be maternal and paternal mutations, respectively. Moreover, population screening and bioinformatic analysis were carried out to determine the effects of these two mutations which revealed that both the mutation were pathogenic. Based on the similar mild phenotype of these two patients, we suggest that OCA3 might be prevalent within the Chinese population.

Robotic Mitral Valve Repair for All Categories of Leaflet Prolapse: Improving Patient Appeal and Advancing Standard of Care

Mayo Clinic Proceedings. Mayo Clinic. Sep, 2011  |  Pubmed ID: 21757782

To characterize the early outcomes of robotic mitral valve (MV) repair using standard open techniques.

A Nuclear Magnetic Resonance Based Approach to Accurate Functional Annotation of Putative Enzymes in the Methanogen Methanosarcina Acetivorans

BMC Genomics. 2011  |  Pubmed ID: 21810209

Correct annotation of function is essential if one is to take full advantage of the vast amounts of genomic sequence data. The accuracy of sequence-based functional annotations is often variable, particularly if the sequence homology to a known function is low. Indeed recent work has shown that even proteins with very high sequence identity can have different folds and functions, and therefore caution is needed in assigning functions by sequence homology in the absence of experimental validation. Experimental methods are therefore needed to efficiently evaluate annotations in a way that complements current high throughput technologies. Here, we describe the use of nuclear magnetic resonance (NMR)-based ligand screening as a tool for testing functional assignments of putative enzymes that may be of variable reliability.

VEGF-C As a Decision-making Biomarker for Selected Patients with Invasive Bladder Cancer Who Underwent Bladder-preserving Radical Surgery

Archives of Medical Research. Jul, 2011  |  Pubmed ID: 21821075

We proved the feasibility of radical transurethral resection in selected patients with muscle-invasive bladder cancer with a minimum follow-up of >5 years. A follow-up schedule was developed based on progression and recurrence during this period.

Thermococcus Kodakarensis Encodes Three MCM Homologs but Only One is Essential

Nucleic Acids Research. Dec, 2011  |  Pubmed ID: 21821658

The minichromosome maintenance (MCM) complex is thought to function as the replicative helicase in archaea and eukaryotes. In eukaryotes, this complex is an assembly of six different but related polypeptides (MCM2-7) but, in most archaea, one MCM protein assembles to form a homohexameric complex. Atypically, the Thermococcus kodakarensis genome encodes three archaeal MCM homologs, here designated MCM1-3, although MCM1 and MCM2 are unusual in having long and unique N-terminal extensions. The results reported establish that MCM2 and MCM3 assemble into homohexamers and exhibit DNA binding, helicase and ATPase activities in vitro typical of archaeal MCMs. In contrast, MCM1 does not form homohexamers and although MCM1 binds DNA and has ATPase activity, it has only minimal helicase activity in vitro. Removal of the N-terminal extension had no detectable effects on MCM1 but increased the helicase activity of MCM2. A T. kodakarensis strain with the genes TK0096 (MCM1) and TK1361 (MCM2) deleted has been constructed that exhibits no detectable defects in growth or viability, but all attempts to delete TK1620 (MCM3) have been unsuccessful arguing that that MCM3 is essential and is likely the replicative helicase in T. kodakarensis. The origins and possible function(s) of the three MCM proteins are discussed.

Aortic Valve Replacement in the Elderly: Determinants of Late Outcome

Circulation. Aug, 2011  |  Pubmed ID: 21824918

Few data exist on long-term outcomes of elderly patients after aortic valve replacement. We evaluated latest follow-up information for patients ≥70 years of age after aortic valve replacement.

MamK, a Bacterial Actin, Forms Dynamic Filaments in Vivo That Are Regulated by the Acidic Proteins MamJ and LimJ

Molecular Microbiology. Oct, 2011  |  Pubmed ID: 21883528

Bacterial actins, in contrast to their eukaryotic counterparts, are highly divergent proteins whose wide-ranging functions are thought to correlate with their evolutionary diversity. One clade, represented by the MamK protein of magnetotactic bacteria, is required for the subcellular organization of magnetosomes, membrane-bound organelles that aid in navigation along the earth's magnetic field. Using a fluorescence recovery after photobleaching assay in Magnetospirillum magneticum AMB-1, we find that, like traditional actins, MamK forms dynamic filaments that require an intact NTPase motif for their turnover in vivo. We also uncover two proteins, MamJ and LimJ, which perform a redundant function to promote the dynamic behaviour of MamK filaments in wild-type cells. The absence of both MamJ and LimJ leads to static filaments, a disrupted magnetosome chain, and an anomalous build-up of cytoskeletal filaments between magnetosomes. Our results suggest that MamK filaments, like eukaryotic actins, are intrinsically stable and rely on regulators for their dynamic behaviour, a feature that stands in contrast to some classes of bacterial actins characterized to date.

Does Lung Ischemia and Reperfusion Have an Impact on Coronary Flow? A Quantitative Coronary Blood-flow Analysis with Inflammatory Cytokine Profile

European Journal of Cardio-thoracic Surgery : Official Journal of the European Association for Cardio-thoracic Surgery. Sep, 2011  |  Pubmed ID: 21900017

Objective: Ischemia-reperfusion (IR) injury remains a major cause of early morbidity and mortality after lung transplantation with poorly documented extrapulmonary repercussions. To determine the hemodynamic effect due to lung IR injury, we performed a quantitative coronary blood-flow analysis in a swine model of in situ lung ischemia and reperfusion. Methods: In 14 healthy pigs, blood flow was measured in the ascending aorta, left anterior descending (LAD), circumflex (Cx), right coronary artery (RCA), right common carotid artery (RCCA), and left internal mammary artery (LIMA), along with left-and right-ventricular pressures (LVP and RVP), aortic pressure (AoP), and pulmonary artery pressure (PAP). Cardiac Troponin (cTn), interleukin 6 and 10 (IL-6 and IL-10), and tumor necrosis factor A (TNF-A) were measured in coronary sinus blood samples. The experimental (IR) group (n=10) underwent 60min of lung ischemia followed by 60min of reperfusion by clamping and releasing the left pulmonary hilum. Simultaneous measurements of all parameters were made at baseline and during IR. The control group (n=4) had similar measurements without lung IR. Results: In the IR group, total coronary flow (TCF=LAD+Cx+RCA blood-flow) decreased precipitously and significantly from baseline (113±41mlmin(-1)) during IR (p<0.05), with the lowest value observed at 60min of reperfusion (-37.1%, p<0.003). Baseline cTn (0.08±0.02ngml(-1)) increased during IR and peaked at 45min of reperfusion (+138%, p<0.001). Baseline IL-6 (9.2±2.17pgml(-1)) increased during IR and peaked at 60min of reperfusion (+228%, p<0.0001). Significant LVP drop at 5min of ischemia (p<0.05) was followed by a slow return to baseline at 45min of ischemia. A second LVP drop occurred at reperfusion (p<0.05) and persisted. Conversely, RVP increased throughout ischemia (p<0.05) and returned toward baseline during reperfusion. Coronary blood flow and hemodynamic profile remained unchanged in the control group. IL-10 and TNF-A remained below the measurable range for both the groups. Conclusions:In situ lung IR has a marked negative impact on coronary blood flow, hemodynamics, and inflammatory profile. In addition, to the best of our knowledge, this is the first study where coronary blood flow is directly measured during lung IR, revealing the associated increased cardiac risk.

Robotic Mitral Valve Repair for All Prolapse Subsets Using Techniques Identical to Open Valvuloplasty: Establishing the Benchmark Against Which Percutaneous Interventions Should Be Judged

The Journal of Thoracic and Cardiovascular Surgery. Nov, 2011  |  Pubmed ID: 21911231

Recent reports have shown that robotic mitral valve repair is effective in treating posterior leaflet disease; however, comparison with trans-sternal (open) valvuloplasty for all prolapse categories has not been performed. Moreover, data from the recently published EVEREST II trial infer that adverse event rates after mitral valve repair for degenerative disease are high. We therefore compared early outcomes of robotic versus open mitral valve repair for patients with mitral valve prolapse.

Replacement of the Infected Composite Aortic Root Prosthesis

The Annals of Thoracic Surgery. Nov, 2011  |  Pubmed ID: 21937018

We sought to evaluate freedom from reinfection after surgery for infected aortic root replacement (ARR) and the impact of use of synthetic material at reoperation.

[Expression of C-fos, C-jun in Hippocampus Under the Model of Transmitting Epileptic Discharge from Brain Tissue to Muscular Tissue on Its Own Skull]

Sheng Wu Yi Xue Gong Cheng Xue Za Zhi = Journal of Biomedical Engineering = Shengwu Yixue Gongchengxue Zazhi. Oct, 2011  |  Pubmed ID: 22097274

The objective of this study was to investigate the changes of the behaviors, EEG and expression of c-fos, c-jun on induced seizure in rats by injecting penicillin after transmitting epileptic discharge from brain tissue to muscular tissue on skull. Eighteen experimental rats were divided into 3 groups, with each 6 rats. Seizure group: 6 acute seizure models were established by injecting penicillin in hippocampus of rats; Transferring group, 6 acute seizure models were established by injecting penicillin in hippocampus of rats, and electrode connected to muscles was planted into epileptic focus of each rat; Control group, 6 rats were only planted electrode in hippocampus without injecting penicillin. Then we observed the changes of behaviors, EEG and expression of C-fos, C-jun in hippocampus with immunohistochemical method. There was no statistic difference in seizure frequency of rat between seizure group and transferring group, but the discharging frequency in EEG of transferring group lowered significantly (P<0.05). The expression of C-fos, C-jun in hippocampus of transferring group rats was significantly lower than that of seizure group (P<0.005). It could be concluded that under the model of transmitting epileptic discharge from brain tissue to muscular tissue on skull, the burst times on EEG electrode decreased, concomitantly with the lower expression of C-fos, C-jun in hippocampus.

Contemporary Results of Open Aortic Arch Surgery

The Journal of Thoracic and Cardiovascular Surgery. Dec, 2011  |  Pubmed ID: 22177097

OBJECTIVES: The success of endovascular therapies for descending thoracic aortic disease has turned attention toward stent graft options for repair of aortic arch aneurysms. Defining the role of such techniques demands understanding of contemporary results of open surgery. METHODS: The outcomes of open arch procedures performed on a single surgical service from July 1, 2001 to August 30, 2010, were examined as defined per The Society of Thoracic Surgeons national database. RESULTS: During the study period, 209 patients (median age, 65 years; range, 26-88) underwent arch operations, of which 159 were elective procedures. In 65 the entire arch was replaced, 22 of whom had portions of the descending thoracic aorta simultaneously replaced via bilateral thoracosternotomy. Antegrade cerebral perfusion was used in 78 patients and retrograde cerebral perfusion in 1. Operative mortality was 2.5% in elective circumstances and 10% in emergency cases (P = .04). The stroke rate was 5.0% when procedures were performed electively and 11.8% when on an emergency basis (P = .11). Procedure-specific mortality rates were 5.5% for elective and 10% for emergency procedures with total arch replacement, and 1.0% for elective and 10% for emergency procedures with hemiarch replacement. Stratified by extent, neurologic event rates were 5.5% for elective and 10% for emergency procedures with total arch and 4.8% for elective and 12.5% for emergency procedures with hemiarch replacement. CONCLUSIONS: Open aortic arch replacement can be performed with low operative mortality and stroke rates, especially in elective circumstances, by a team with particular focus on the procedure. The results of novel endovascular therapies should be benchmarked against contemporary open series performed in such a setting.

Effect of the Combination of Music and Nature Sounds on Pain and Anxiety in Cardiac Surgical Patients: a Randomized Study

Alternative Therapies in Health and Medicine. Jul-Aug, 2011  |  Pubmed ID: 22314630

Background Postoperative pain and anxiety are common in cardiac surgery patients. Studies have suggested that music can decrease anxiety in hospitalized patients. Primary Study Objective This study focused on the efficacy and feasibility of special music, which included nature sounds, for pain and anxiety. Methods/Design In this randomized controlled trial, postoperative cardiovascular surgery patients were randomly assigned to a music group to receive 20 minutes of standard postoperative care and music twice daily on postoperative days 2 through 4 or to a control group to receive 20 minutes of standard care with a quiet resting period twice daily on postoperative days 2 through 4. Setting Cardiovascular surgical unit of Saint Marys Hospital, Rochester, Minnesota. Participants One hundred patients completed the study (music group, n = 49; control group, n = 51). Intervention The music was delivered through CD players in the patients' rooms. Primary Outcome Measures Pain, anxiety, satisfaction, and relaxation were evaluated from visual analog scales. Results Data showed a significant decrease in mean (SD) pain scores after the second session of day 2 for the music group (change, ?1.4 [1.4]) compared with the control group (change, ?0.4 [1.4]) (P = .001). Mean relaxation scores improved more at the first session of day 2 for the music group (change, 1.9 [2.7]) compared with the control group (change, 0.3 [2.9]) (P = .03). The music group also showed lower anxiety and increased satisfaction overall, but these differences were not statistically significant. No major barriers to using the therapy were identified. Conclusion Recorded music and nature sounds can be integrated into the postoperative care of cardiovascular surgery patients. The recordings may provide an additional means for addressing common symptoms of pain and anxiety while providing a means of relaxation for these patients.

Single-patch, 2-patch, and Caval Division Techniques for Repair of Partial Anomalous Pulmonary Venous Connections: Does It Matter?

The Journal of Thoracic and Cardiovascular Surgery. Feb, 2012  |  Pubmed ID: 22325328

OBJECTIVE: We reviewed our experience with the single-patch, 2-patch, and caval division techniques for repair of a partial anomalous pulmonary venous connection. METHODS: From 1990 to 2009, 124 patients (65 male patients; 52%) were identified. The single-patch, two-patch, and caval division technique was used in 60 (49%), 24 (19%), and 40 (32%) patients, respectively. The median age was 34.5 years (range, 1-76). Of the 124 patients, 99 (80%) had an atrial septal defect. RESULTS: There was no early mortality. Survival at 5, 10, and 15 years was 99%, 95%, and 82%, respectively. One patient (3%) in the caval division group required early reoperation for superior vena caval obstruction. All patients, exept for 3, had regained sinus rhythm at discharge. Late superior vena caval obstruction developed in 3 patients (7.5%) in the caval division group, 3 (5%) in the single-patch group, and 1 (4%) in the 2-patch group (Р = .5). Two patients underwent reoperation for late pulmonary vein obstruction: 1 (2%) in the single-patch group and 1 (4%) in the 2-patch group (P = .08). A permanent pacemaker was required in 3 patients (3%). CONCLUSIONS: Surgical treatment of partial anomalous pulmonary venous connections is associated with excellent outcomes. The overall incidence of late superior vena caval or pulmonary vein stenosis is low. Although not significant, the 2-patch technique might be associated with a greater incidence of sinus node dysfunction and late pulmonary venous stenosis. The late development of superior vena caval obstruction is a concern with all techniques, necessitating close follow-up.

Changes in Renal Function After Implantation of Continuous-flow Left Ventricular Assist Devices

Journal of the American College of Cardiology. Jan, 2012  |  Pubmed ID: 22192665

The aim of this study was to determine renal outcomes after left ventricular assist device (LVAD) implantation.

Evaluation of Susceptibility Locus for Response to Interferon-α Based Therapy in Chronic Hepatitis B Patients in Chinese

Antiviral Research. Feb, 2012  |  Pubmed ID: 22209781

In 2009, three independent genome-wide association studies reported that genetic variation in the interleukin 28B gene to be associated with the response to interferon-α/ribavirin therapy in hepatitis C virus genotype 1 infected patients. We carried out the present study to assess whether such polymorphisms also affect the therapy effect of another interferon-α responsive illness as chronic hepatitis B. Five hundred and twelve interferon-α treatment-naïve HBeAg seropositive chronic hepatitis B patients were enrolled in the present retrospective nested case-control study. All patients received PEG-IFN-α-2a based treatment and were examined for the therapy efficacy. SNP rs8099917 was genotyped using the MassArray system (Sequenom). Interestingly, the frequency of G allele of rs8099917 was significantly higher in response group than in non response group (8.3% vs. 3.9%, p=0.003, OR=0.44, 95%CI=0.25-0.79). The genotype distributions of this SNP also differed significantly between two groups (p=0.003). Our study suggested that the G allele of rs8099917 was associated with higher rate of response in HBeAg seropositive chronic hepatitis B patients treated with interferon α.

Polymorphisms in the Potential Functional Regions of the TGF-β 1 and TGF-β Receptor Genes and Disease Susceptibility in HBV-related Hepatocellular Carcinoma Patients

Molecular Carcinogenesis. Jan, 2012  |  Pubmed ID: 22290546

Hepatocellular carcinoma (HCC) is a disease of multiple etiologies caused by the accumulation of genetic and epigenetic defects. Current evidence indicates that the transforming growth factor beta (TGF-β) signaling pathway has a significant impact on different cellular process. Members of the TGF-β superfamily (TGF-β1, the type I TGF-β receptor [TβRI], type II TGF-β receptor [TβRII], and type III TGF-β receptor]) play an important role in tumorigenesis. Numerous studies show that genetic polymorphisms in TGF-β superfamily genes are associated with HCC in East Asian populations. We studied 16 single nucleotide polymorphisms (SNPs) in four genes (TGF-β1, TβRI, TβRII, and TβRIII) to examine their associations with hepatocarcinogenesis. A total of 1228 Chinese Han participants were enrolled in the study (881 control participants who were negative for all hepatitis B virus [HBV] serum markers and 347 case participants with HBV-related HCC). Genotyping was conducted using the TaqMan method. The results showed that the frequency of the rs1805110 T allele was significantly higher in the case group than in the control group (P = 0.034). After stratification, the results for rs1805110 remained significant in male participants (P = 0.005), but there was no statistical difference in females. In males, the frequency of the C-C-G-C-A haplotype resulting from SNPs rs1805110, rs2810904, rs1805112, rs284878, and rs1804506 in TβRIII was significantly lower in the case group than in the control group (P = 0.001), whereas the reverse was true for the T-C-G-C-A haplotype (P = 0.036). We conclude that the rs1805110T allele is associated with susceptibility to HBV-related HCC in males. © 2012 Wiley Periodicals, Inc.

The V Protein of Mumps Virus Plays a Critical Role in Pathogenesis

Journal of Virology. Feb, 2012  |  Pubmed ID: 22090137

Mumps virus (MuV) causes an acute infection in humans characterized by a wide array of symptoms ranging from relatively mild manifestations, such as parotitis, to more-severe complications, such as meningitis and encephalitis. Widespread mumps vaccination has reduced mumps incidence dramatically; however, outbreaks still occur in vaccinated populations. The V protein of MuV, when expressed in cell culture, blocks interferon (IFN) expression and signaling and interleukin-6 (IL-6) signaling. In this work, we generated a recombinant MuV incapable of expressing the V protein (rMuVΔV). The rescued MuV was derived from a clinical wild-type isolate from a recent outbreak in the United States (MuV(Iowa/US/06), G genotype). Analysis of the virus confirmed the roles of V protein in blocking IFN expression and signaling and IL-6 signaling. We also found that the rMuV(Iowa/US/06)ΔV virus induced high levels of IL-6 expression in vitro, suggesting that V plays a role in reducing IL-6 expression. In vivo, the rMuV(Iowa/US/06)ΔV virus was highly attenuated, indicating that the V protein plays an essential role in viral virulence.

A SNX10/V-ATPase Pathway Regulates Ciliogenesis in Vitro and in Vivo

Cell Research. Feb, 2012  |  Pubmed ID: 21844891

Sorting nexins (SNXs) are phosphoinositide-binding proteins implicated in the sorting of various membrane proteins in vitro, but the in vivo functions of them remain largely unknown. We reported previously that SNX10 is a unique member of the SNX family genes in that it has vacuolation activity in cells. We investigate the biological function of SNX10 by loss-of-function assay in this study and demonstrate that SNX10 is required for the formation of primary cilia in cultured cells. In zebrafish, SNX10 is involved in ciliogenesis in the Kupffer's vesicle and essential for left-right patterning of visceral organs. Mechanistically, SNX10 interacts with V-ATPase complex and targets it to the centrosome where ciliogenesis is initiated. Like SNX10, V-ATPase regulates ciliogenesis in vitro and in vivo and does so synergistically with SNX10. We further discover that SNX10 and V-ATPase regulate the ciliary trafficking of Rab8a, which is a critical regulator of ciliary membrane extension. These results identify an SNX10/V-ATPase-regulated vesicular trafficking pathway that is crucial for ciliogenesis, and reveal that SNX10/V-ATPase, through the regulation of cilia formation in various organs, play an essential role during early embryonic development.

High Glucose-induced Hypertrophy of Mesangial Cells is Reversed by Connexin43 Overexpression Via PTEN/Akt/mTOR Signaling

Nephrology, Dialysis, Transplantation : Official Publication of the European Dialysis and Transplant Association - European Renal Association. Jan, 2012  |  Pubmed ID: 21633095

Hypertrophy of glomerular mesangial cells (GMC) is one of the earliest pathological abnormalities in diabetic nephropathy, which correlates with eventual glomerulosclerosis. We have previously proved that this hypertrophy is mediated by downregulation of connexin43 (Cx43) and dysfunction of gap junctional intercellular communication, but the mechanism involved is still unclear. This study aims to investigate whether PTEN/Akt/mammalian target of rapamycin (mTOR) was involved as the downstream molecular signaling of Cx43 in regulating high glucose-induced GMC hypertrophy.