bluuebunny/crossref_metadata_2025_split huggingface dataset:
Identification of an AP‐2 element in the ‐323 to ‐285 region of the TNF‐α gene
Authors: Kroeger Karen M. , Abraham Lawrence J. 
Published: July 2007

AbstractWe have identified a region of the human TNF‐α promoter between nucleotides ‐323 and ‐285, capable of influencing transcriptional activity. This region encompasses the ‐308 polymorphism and contains a 10 bp sequence homologous to the consensus binding site of activator protein‐2 (AP‐2). Protein complexes derived from U937 and Jurkat cells were found to bind to this element. Competitive EMSA using a consensus AP‐2 oligonucleotide indicated that AP‐2 may be involved. Functional assays demonstrate that this region can repress activity of a heterologous promoter in the Jurkat T‐cell line, but act as an inducible enhancer of transcription in U937 cells.
Expression of cecropin CMIV fusion protein in E. coli under T7 promoter
Authors: Xie Wei , Qiu Qifeng , Wu Haihong , Xu Xianxiu 
Published: July 2007

AbstractA synthetic gene coding for cecropin CMIV from Bombyx mori was cloned into plasmid pEZZ318 and fused to a DNA segment encoding the signal peptide of Staphylococcal protein A and IgG binding domain. The fusion gene was then subcloned into expression vector pET11c under the control of T7 promoter and expressed in E.coli. The fusion protein did not exhibit any antibacterial activity either in cell lysate or in medium. After cleavage from the fusion protein with CNBr the biological activity of recombinant cecropin CMIV was obtained.
Enhancement by F‐actin of MgATP‐dependent dopamine uptake into isolated chromaffin granules
Authors: Morita Kyoji , Tomares Stuart M. , Pollard Harvey B. 
Published: July 2007

AbstractInteraction of chromaffin granules with the microfilament network in the cytoplasmic space of adrenal medullary cells has been proposed, but the influence of chromaffin granule‐microfilament interaction on granule function has not yet been studied. In the present study, the effect of F‐actin on dopamine uptake into isolated chromaffin granules was examined. The MgATP‐dependent component of dopamine uptake was significantly enhanced by F‐actin. In contrast, catecholamine efflux from the granules was not significantly altered by F‐actin under the conditions in which the enhancement of MgATP‐dependent dopamine uptake was observed. These findings suggest that the interaction of chromaffin granules with the microfilament network results in the enhancement of MgATP‐dependent dopamine uptake into the granules, thus modulating the accumulation and storage of catecholamines in the adrenal medullary cell.
C‐terminal peptide of streptokinase, Met369‐Pro373, is important in plasminogen activation
Authors: Kim Il Chul , Kim Jang Seong , Lee Si Hyoung , Byun Si Myung 
Published: July 2007

AbstractStreptokinase(SK), a plasminogen activator, is known to have multi‐domain structure. The function of the C‐terminal region of streptokinase was investigated with SK mutants constructed by truncating 26, 33, 37, 40, 41, 46, 47, 70 or 97 amino acid residues from the C‐terminus. The truncated SKs were expressed in E.coli and purified. The 41 residue deletion (SKP373) from the C‐terminus had not effect on the plasminogen activation activity. However, the deletion of 46 amino acid residues (SKP368) resulted in the dramatic reduction of the plasminogen activation efficiency. The result suggests that the C‐terminal peptide from Met369 to Pro373 of SK may play an important role on the plasminogen activation.
Identification of macrophage migration inhibitory factor in synovial membranes of loosened total joint replacement
Authors: Suzuki Kouji , Onodera Shin , Matsuno Takeo , Kaneda Kiyoshi , Nishihira Jun 
Published: July 2007

AbstractEndosteal bone resorption often occurs around loosened total joint replacements. In the process of the loosening, macrophages play an important role by releasing cytokines such as interleukin‐1, tumor necrosis factor‐α and prostaglandin E2. In this study, we investigated the involvement of macrophage migration inhibitory factor (MIF) in the pathological state of the loosening of a total hip replacement. Interface membranes were harvested from bone‐cement or bone‐implant interfaces of two patients during revision hip surgeries. The tissues were immunohistochemically examined with a polyclonal antibody against human recombinant MIF. This study detected MIF in the cytoplasm of the macrophages in all the tissues tested, though it was not detected in that of interstitial cells. The expression of MIF mRNA in the membrane was also examined by reverse transcription polymerase chain reaction, which demonstrated that expression of the MIF mRNA in the interface membranes was higher than that of the normal synovium. Considering these results, it is suggested that MIF is one of important cytokines mediating the inflammatory process during loosening of total joint replacements.
Structural and functional roles of the amino‐terminal region and collagen‐like domain of human serum mannan‐binding protein
Authors: Ma Yong , Yokota Yasunori , Kozutsumi Yasunori , Kawasaki Toshisuke 
Published: July 2007

AbstractThe serum mannan‐binding protein (S‐MBP) is a Ca2+‐dependent C‐type animal lectin specific for mannose and N‐acetylglucosamine, which plays an important role in first‐line host defense. To study the structure and function relationship of the lectin, a full‐length human S‐MBPcDNA was expressed in Sf9 insect cells using a baculovirus expression system, and a cDNA encoding the carbohydrate recognition domain (CRD) of human S‐MBP was expressed in E. coli. The properties of the recombinant S‐MBP and recombinant S‐MBP‐CRD were compared with those of the native human S‐MBP and the CRD of the native S‐MBP. The results indicated that functional human S‐MBP can be successfully expressed in Sf9 cells and functional S‐MBP‐CRD in E. coli. In addition, the amino‐terminal region and collagen‐like domain are required for higher oligomer formation and play important roles in complement activation.
Identification and characterization of an RNA specific primer for human tumor necrosis factor receptor I (TNFR‐I)
Authors: Kuo Kou‐Wha , Leung Maria FKL , Leung Wai‐Choi 
Published: July 2007

AbstractAn RNA specific primer amplified TNFR‐I cDNA from cellular RNA by single‐tube (ST) RT‐PCR without interfering with the presence of genomic DNA of cell was identified. The primer detected TNFR‐I from 1 ng of cellular RNA by ST RT‐PCR, and was not cross‐reacted with TNFR‐II despite of the homologous sequence between TNFR‐I and ‐II. In addition, the mechanism of the RNA specificity of the primer was investigated. An intron of 0.43 kb was inserted in the 5′‐925 of TNFR‐I mRNA sequence. The result corresponded with the previous determination of TNFR‐I gene structure. Thus, it appears that the RNA specificity of the primer might be resulted from the antisense primer hybridizing to exon‐intron junction. Owing to its high sensitivity and specificity to TNFR‐I RNA, the RNA specific primer may be potentially utilized for the determination of human TNFR‐I gene expression by in situ RT‐PCR.
Identification and cloning of a novel type I sering / threonine kinase receptor of the TGF‐β / BMP superfamily in rat prostate
Authors: Kang Yong , Reddi A. Hari 
Published: July 2007

AbstractA cDNA clone encoding a new member of the type I serine / threonine kinase receptors of the TGF‐β / BMP superfamily was isolated from a rat prostate cDNA library. The predicted sequence of the encoded protein is 493 amino acids with a molecular weight of 55 kDa. It contains a putative cysteine rich extracellular domain, a transmembrane domain, a conserved type I receptor GS domain and a cytoplasmic serine / threonine kinase domain. The kinase domain shares 80% homology with that of ALK‐4 and ALK‐5 whose ligands are activin and TGF‐β. Northern blot analysis revealed that this prostate serine / threonine kinase receptor I (PSKR‐I) is highly expressed in rat dorsolateral prostate, coagulating gland (anterior prostate), cerebellum and adipose tissue.
Is protein kinase C alpha (PKCα) involved in vasopressin‐induced effects on LLC‐PK1 pig kidney cells?
Authors: Dibas Adnan , Mia Abdul J. , Yorio Thomas 
Published: July 2007

AbstractThe involvement of protein kinase C (PKC) in vasopressin‐induced effects on renal water reabsorption is still unresolved. Activation of PKC can be detected by its translocation from the cytosol (C) to the plasma membrane (PM). In LLC‐PK1 cells, the redistribution of PKCα, a predominant isoform of PKC detected, was studied utilizing western blotting after stimulation with vasopressin. Vasopressin (100 mU/ml) failed to induce a translocation of PKCα from the C to the PM. By contrast, phorbol myristate acetate (PMA, 200 nM), a potent activator of PKC, induced a relocalization of PKCα from the C to the PM. After 2 hours of treatment of cells with PMA, PKCα was predominantly detected in the PM and absent from the C. These results suggest that the signal transduction pathway of vasopressin in LLC‐PK1 cells does not involve PKCα activation and translocation.
The effect of NG‐monomethyl‐L‐arginine(LNMMA), an no‐synthase blocker on the survival of intracellular BCG within human monocyte‐derived macrophages
Authors: Fazal Nadeem 
Published: July 2007

AbstractHuman peripheral blood monocytes are permissive for the growth of Mycobacterium bovis (BCG), but the fate of nonpathogenic Mycobacterium bovis in these cells is not clearly known. Both oxidative and nonoxidative pathways have been implicated in killing of intracellular mycobacteria. Since human monocytes are not so far confirmed to release Reaction Nitrogen Intermediates (RNI), we tried indirect approach to inhibit the production of NO by the addition of NG‐monomethyl‐L‐arginine (LNMMA)‐an NO synthase blocker, in the infected cell cultures. In our studies adherent human peripheral blood monocytes were found to be permissive for the growth of Mycobacterium bovis, as measured by [3H] uridine uptake and confirmed by Colony Forming Unit (CFU) estimate. The killing of Mycobacterium bovis was not blocked by LNMMA, suggesting that it was not due to the production of Reactive Nitrogen Intermediates. Culture of the monocyte‐derived macrophages for 1 to 14 days before infection had no effect on the fate of Mycobacterium bovis, and no nitric oxide was detected in the culture supernatants of these infected cell cultures. Graded doses of sodium nitrite at physiological concentrations however failed to affect cell free cultures of Mycobacterium bovis. These findings suggest that the addition of NG‐monomethyl‐L‐arginine (LNMMA), an antagonist of L‐arginine oxidation and inhibitor of NO production, had no effect on the fate of Mycobacterium bovis in human monocytes and macrophages.
Salinity and osmotic stress‐regulated proteins in cowpea Rhizobium 4a (peanut isolate)
Authors: Wankhade Sharad , Apte S. K. , Rao K. Koteswara 
Published: July 2007

AbstractThe effect of salinity and osmotic stress on protein synthesis was studied in cowpea Rhizobium 4a. Osmotic component of salinity stress has been shown to induce ten proteins in a salt tolerant/ osmosensitive cowpea Rhizobium. 4a (groundnut isolate). Of these seven polypeptides were induced only by salt/osmotic stress while two were induced by heat shock. The results demonstrate a commonality as well as stress specificity of protein synthesis regulation.
Dopaquinone hydroxylation through topaquinone cofactor in copper amine oxidases: A simplified chemical model
Authors: Rinaldi Andrea C. , Rescigno Antonio , Sollai Francesca , Soddu Giulia , Curreli Nicoletta , Rinaldi Augusto , Finazzi‐Agrò Alessandro , Sanjust Enrico 
Published: July 2007

AbstractA simple model, 4‐tert‐butyl‐1,2‐benzoquinone, was chosen to study the hydroxylation step of the tyrosine‐derived Dopaquinone residue at the active site of copper amine oxidases in the self‐catalytic generation of the Topaquinone cofactor. This hydroxylation step was studied both in the presence and absence of free copper(II), and was found to be dependent on pH value but not on the presence of metal ions. It is therefore proposed that, hydroxide ion and not water should be the true reactive species in this key biosynthetic step of the Topaquinone cofactor, and that the active site Cu2+ is implied, at this point of cofactor biosynthesis, in the quinonisation of Topa rather than in the hydroxylation of Dopaquinone.
Specific mitochondrial DNA deletions in canine myocardial ischemia
Authors: Marin‐Garcia Jose , Goldenthal Michael J. , Ananthakrishnan Radha , Mirvis David 
Published: July 2007

AbstractThe effect of myocardial ischemia on mitochondrial DNA (mtDNA) structure and the presence of specific mtDNA deletions was determined using the model of Ameroid constriction of canine coronary arteries. The incidence of specific deletions was high in both the endocardial and epicardial tissues perfused by the occluded vessel as compared to myocardial tissues perfused by the unconstricted vessels. Our results show that specific mtDNA deletions similar to the 5 kb and 7.4 kb human mtDNA deletions occur following canine myocardial ischemia. However the presence of these deletions did not correlate with specific mitochondrial enzyme defects.
Effect of insulin on DNA synthesis and kinetic parameters of thymidine kinase during liver regeneration
Authors: Mola P. Waliaula , Sudha B. , Paulose C. S. 
Published: July 2007

AbstractThe effect of insulin on cell proliferation in vivo has been studied in hepatectomised streptozotocin‐ diabetic rats. The extent of cell proliferation in sham and hepatectomized‐ control, diabetic and insulin treated rats were monitored by determining DNA content and [3H]thymidine incorporation into DNA. The kinetic parameters of thymidine kinase a regulatory enzyme for DNA synthesis was also studied in these groups. The rate of DNA synthesis in liver of streptozotocin ‐diabetic rats was significantly higher 24 hrs post‐hepatectomy compared to control and insulin treated diabetic groups. Kinetic studies of thymidine kinase revealed that there was no change in the Michaelis ‐Menten constant (Km) whereas maximum velocity (Vmax) was elevated in the diabetic hepatectomized groups compared to control and insulin treated hepatectomized groups. Thus our study elucidates the role of insulin in thymidine kinase activity and DNA synthesis.
Subcellular redistribution of hsp72 protein during cisplatin‐induced apoptosis in HeLa cells
Authors: Meléndez‐Zajgla Jorge , García Claudia , Maldonado Vilma 
Published: July 2007

AbstractExposure of HeLa cells to cisplatin results in the activation of apoptotic cell death. This drug induces DNA damage and generates reactive oxygen intermediates. Since cisplatin is highly reactive and binds to diverse proteins, it could create abnormal protein structures or nonspecific aggregates. For these reasons, we analyzed the expression and subcelullar distribution of hsp72, a heat‐shock protein that enables cells under stress to cope with damaged proteins. We did not observe any changes in the expression of hsp72 protein, although, by immunofluorescence studies, we detected a dramatic redistribution of the protein. These results and its probable relevance in the drug‐induced apoptotic phenomenon are discussed.
Further evidence for the mitogenic action of urinary trypsin inhibitor
Authors: Manilal Sushila B. , Scott G. Kenneth 
Published: July 2007

AbstractWe have previously shown that urinary trypsin inhibitor (UTI), also known as bikunin, was mitogenic for human fibroblasts at low concentrations, and growth‐inhibitory at higher concentrations, and have identified high‐ and low‐affinity cellular binding sites for this protein. We have now investigated fibroblast proteins which interact with bikunin. Bikunin binds to proteins of about 50K and 250K. The simplest interpretation is that the 50K protein may be a proteinase which is also the low‐affinity bikunin binding site, involved in growth inhibition, and that the larger protein may be responsible for the mitogenic response to bikunin. Inhibitors of intracellular calcium mobilisation also inhibit the mitogenic response to bikunin, and, by the measurement of the efflux of pre‐loaded 45Ca2+, bikunin at mitogenic concentrations can be shown to stimulate calcium mobilisation.
Synchronized induction in bFGF and Cdk2 during gastric ulcer healing
Authors: Slomiany B. L. , Piotrowski J. , Slomiany A. 
Published: July 2007

AbstractThe processes of gastric mucosal repair are regulated by growth factors triggering the signals for cell cycle progression. We examined the expression of gastric mucosal basic fibroblast factor (bFGF) and cyclin dependent kinase (p34Cdk2) at different stages of chronic ulcer healing. The results of assays revealed that ulcer healing was accompanied by an increase in mucosal expression of bFGF and Cdk2. A significant induction in bFGF and Cdk2 occurred by the 2nd day of ulcer healing, reached the respective maximums of 2.3‐ and 2.2‐fold increase by the 4th day of healing, and remained still elevated (20%) on the 14th day when the ulcer was essentially healed. The results demonstrate a synchronized induction in bFGF and cell cycle regulatory kinase, Cdk2, with ulcer healing.
Amino acid sequence of catechol 1,2‐dioxygenase (pyrocatechase) isozyme αα from Pseudomonas arvilla C‐1
Authors: Nakai Chieko , Yamazaki Gyohzan , Kagamiyama Hiroyuki , Nozaki Mitsuhiro 
Published: July 2007

AbstractCatechol 1,2‐dioxygenase catalyzes the oxygenative ring cleavage of catechol to form cis,cis‐muconic acid, and contains a ferric form of iron as its sole cofactor. Pseudomonas arvilla C‐1 contains three isozymes of the enzyme, αα, αβ, and ββ [C. Nakai et al. (1990) J. Biol. Chem. 265, 660‐665]. We have determined the amino acid sequence of the αα isozyme by direct analysis. The sequence shared 77% homology with isozyme ββ, and had conserved tyrosyl and histidyl residues which are thought to be involved in the binding of ferric ion.
Purification and properties of chitinase from cabbage stems with roots
Authors: Chang Chen‐Tien , Hsueh Yi‐Ling , Sung Hsien‐Yi 
Published: July 2007

AbstractChitinase has been purified from the extract of cabbage stems with roots through successive steps of ammonium sulfate fractionation, Sephadex G‐75 gel filtration, chromatofocusing and Sephacryl S‐200 HR gel filtration. By these steps, the purity of the enzyme increased by 63 fold and the recovery of the enzyme activity was 18%. The purified enzyme was homogeneous when analyzed by SDS‐PAGE#. It showed an optimal pH of 6 and optimal temperature of 60°C for hydrolysis of ethylene glycol chitin (EGC). The molecular mass of the enzyme was 41 kDa, as determined by SDS‐PAGE. Heavy metal ions (1.5 mM) Ag+, Hg2+ and Fe2+, and chemical modification agents NAI (1 mM), NBS (0.5 mM) and CHD (0.5 mM) significantly or completely inhibited the activity of the enzyme. Substrate EGC at high concentrations also inhibited the activity. BSA (0.05%), Triton X‐100 (0.5%) and glycerol (50%) provided significant protection of the enzyme from freezing inactivation.
Cloning and characterization of cDNA for human adenylate kinase 2A
Authors: Lee Younghee , Kim Jae Wha , Lee In Ae , Kang Ho bum , Choe Yong‐Kyung , Lee Hee gu , Lim Jong‐Seok , Kim Hyo Joon , Park Chankyu , Choe In Seong 
Published: July 2007

AbstractWe have isolated and characterized a cDNA clone encoding human adenylate kinase 2A (AK2A) from cDNA libraries of fetal liver origin. The complete nucleotide sequence of the cloned 889 nucleotide cDNA fragment indicated that the deduced gene product of human AK2A is composed of 239 amino acids with a molecular weight of 26 kD. Comparison of these nucleotide and amino acid sequences with the corresponding sequences of bovine AK2A and rat AK2 revealed a high degree of conservation. It showed 91% and 98% homologies in DNA and amino acid sequences, respectively, with bovine AK2A throughout the full open reading frame. RNA blot analysis revealed that three species of mRNA were present with approximate sizes of 3.4, 2.1, and 1.0 kb. This gene was expressed in E. coli cells and the recombinant protein was enzymatically active.
Regulation of cholesterol synthesis and esterification in primary cultures of macrophages following uptake of Chylomicron remnants
Authors: Yu Kenneth C. ‐W. , Mamo John C. L. 
Published: July 2007

AbstractThe effects of chylomicron remnants (CR), β‐very‐low‐density‐lipoproteins (β‐VLDL) and low‐density‐lipoproteins (LDL) on intracellular cholesterol synthesis and esterification in primary rabbit macrophages was determined by assaying for HMG‐CoA reductase activity and cholesterol esterification. At physiological cholesterol concentrations, both CR and LDL inhibited cholesterol synthesis by almost 60% while β‐VLDL was less potent achieving only 30% inhibition. Cholesterol esterification rates were increased four‐fold by CR and LDL, whereas β‐VLDL increased esterification 14 times above controls. Qualitatively, the effect of CR on cholesterol synthesis and esterification in rabbit macrophages differs from observations in transformed macrophage cells. Quantitatively, the enhanced rates of cholesterol esterification and weak inhibition of cholesterol synthesis following β‐VLDL uptake may explain why this lipoprotein rapidly induces foam cell formation in vitro.
Genetic defect of the sodium pump‐defective mutant nap‐1 from the marine Vibrio alginolyticus
Authors: Hayashi Maki , Fujii Junko , Unemoto Tsutomu 
Published: July 2007

AbstractThe marine bacterium, Vibrio alginolyticus, has a respiratory chain‐linked Na+‐translocating NADH‐quinone reductase (NQR). Among several mutant cells defective in Na+ pump activity, Nap1 was a very stable mutant and a spontaneous revertant could not be isolated from Nap1. Using genetic irformation from the recently sequenced nqr operon, the genetic defects in Nap1 were examined, and the sodium pump‐defective mutant Napl was found to be caused by the insertion of a 1.2 kbp DNA fragment into the C‐terminal region of nqr6 gene.
Decrease in platelet, erythrocyte and lymphocyte acetylcholinesterase activities due to the presence of protease inhibitors in the storage buffer
Authors: Martín‐Valmaseda Eva M. , Sánchez‐Yagüe Jesús , Marcos Rocío , Llanillo Marcial 
Published: July 2007

AbstractThe effect of the presence of protease inhibit0rs in the storage buffer on acetylcholinesterase (AChE) activity was studied in crude membrane preparations from sheep platelets before and after solubilization of the membranes with Triton X‐100. Although sensitive to the action of trypsin, the biological activity of AChE remained unchanged for as long as 6 days at 4°C in a protease‐inhibitor‐free medium. At 10‐5 M final concentration PMSF reduced AChE activity to 50% after 24 hours of storage. This reduction was abolished in mixtures in which PMSF was present together with 3 mM EGTA, 5 mM EDTA and 0.i mg/mL trypsin inhibitor. Nevertheless, under these conditions, storage periods longer than 24 hours still drastically reduced AChE activity. A mixture of EDTA, EGTA and trypsin inhibitor also produced a decrease in AChE activity after 24 hours. A more complex cocktail of inhibitors including several commonly used peptides decreased AChE activity only if PMSF or EDTA were present in the mixtures. Similar results were obtained with sheep erythrocytes or lymphocytes, and bovine erythrocytes.
Expression and reconstitution of Calcineurin A and B subunits
Authors: Wei Qun , Lee Ernest Y. C. 
Published: July 2007

AbstractCalcineurin consists of two subunits, a catalytic A subunit of 60 kDa and a regulatory B subunit of 19 kDa. Both the A and B subunits of rat brain calcineurin were expressed in E. coli. The B subunit was readily overexpressed in the pET‐21 a vector with yields of >70 mg of purified B subunit per 1 culture, representing >17% of the soluble E. coli protein. About 8 mg of purified A subunit was obtained. The enzyme activities of the A‐subunit and the reconstituted AB complex were found to be comparable to that of the bovine brain enzyme. The reconstitution of the AB complex was studied and shown to be rapid.