bluuebunny/crossref_metadata_2025 huggingface dataset:
Expression of matrix metalloproteinase‐9 (gelatinase B) in gouty arthritis and stimulation of MMP‐9 by urate crystals in macrophages
Authors: Hsieh Ming‐Shium , Ho Hsin‐Chiu , Chou Der‐Tsay , Pan Shiann , Liang Yu‐Chih , Hsieh Tsu‐Yi , Lan Joung‐Liang , Tsai Shu‐Huei
Published: July 2003

To investigate the relevance of gelatinase‐B (matrix metalloproteinase 9, MMP‐9) in gouty arthritis (GA), we tested the occurrence of MMP‐9 in GA patients and cell culture system. Gelatinolytic activity in the synovial fluid (SF) of patients with different kinds of arthritis was assessed by gelatin zymography. A predominant 92‐kDa MMP‐9 gelatinolytic activity was evident in rheumatoid arthritis (RA) and GA samples, but no activity was observed in osteoarthritis (OA) samples. Among the 53 SF samples (9 RA, 24 GA, and 20 OA) analyzed for MMP‐9 and tissue inhibitor of metalloproteinase (TIMP‐1) antigen levels by ELISA, MMP‐9 antigen levels were elevated tenfold in GA SF compared with OA SF. In addition, GA synovial tissue extracts revealed elevated levels of MMP‐9 expression as compared to OA tissue extracts by Western blot and RT‐PCR analysis. Immunohistochemical studies demonstrated that MMP‐9 immunoreactivity was more intense in GA than in OA synovial tissues. Furthermore, macrophages activation by gouty crystals in vitro was examined. Crystals stimulated MMP‐9 gene expression in macrophage cell line and such stimulation was suppressed by PD98059. These findings suggest that the abnormal production of MMP‐9 by macrophages is a reflection of the pathological conditions in joints of patients with GA, and that the activation of MMP‐9 in the joint is known to play an important role in joint disease. J. Cell. Biochem. 89: 791–799, 2003. © 2003 Wiley‐Liss, Inc.
1,25‐dihydroxyvitamin D3 inhibits ultraviolet B‐induced apoptosis, Jun kinase activation, and interleukin‐6 production in primary human keratinocytes
Authors: De Haes Petra , Garmyn Marjan , Degreef Hugo , Vantieghem Katleen , Bouillon Roger , Segaert Siegfried
Published: July 2003

We investigated the capacity of 1,25‐dihydroxyvitamin D3 [1,25(OH)2D3] to protect human keratinocytes against the hazardous effects of ultraviolet B (UVB)‐irradiation, recognized as the most important etiological factor in the development of skin cancer. Cytoprotective effects of 1,25(OH)2D3 on UVB‐irradiated keratinocytes were seen morphologically and quantified using a colorimetric survival assay. Moreover, 1,25(OH)2D3 suppressed UVB‐induced apoptotic cell death. An ELISA, detecting DNA‐fragmentation, demonstrated that pretreatment of keratinocytes with 1,25(OH)2D3 1 μM for 24 h reduced UVB‐stimulated apoptosis by 55–70%. This suppression required pharmacological concentrations 1,25(OH)2D3 and a preincubation period of several hours. In addition, 1,25(OH)2D3 also inhibited mitochondrial cytochrome c release (90%), a hallmark event of UVB‐induced apoptosis. Furthermore, we demonstrated that 1,25(OH)2D3 reduced two important mediators of the UV‐response, namely, c‐Jun‐NH2‐terminal kinase (JNK) activation and interleukin‐6 (IL‐6) production. As shown by Western blotting, pretreatment of keratinocytes with 1,25(OH)2D3 1 μM diminished UVB‐stimulated JNK activation with more than 30%. 1,25(OH)2D3 treatment (1 μM) reduced UVB‐induced IL‐6 mRNA expression and secretion with 75–90%. Taken together, these findings suggest the existence of a photoprotective effect of active vitamin D3 and create new perspectives for the pharmacological use of active vitamin D compounds in the prevention of UVB‐induced skin damage and carcinogenesis. J. Cell. Biochem. 89: 663–673, 2003. © 2003 Wiley‐Liss, Inc.
Effect of glucose concentration on activation of the ASK1–SEK1–JNK1 signal transduction pathway
Authors: Song Jae J. , Lee Yong J.
Published: July 2003

Recently, acute total glucose deprivation has been shown to cause activation of ASK1–MEK–MAPK signal transduction and dissociation of glutaredoxin (GRX) from apoptosis signal‐regulating kinase 1 (ASK1). In this study, we investigated whether clinically relevant concentrations (0.01–0.1 mM) of glucose promote ASK1 activation. We observed that a prominent activation of JNK1 occurred at a glucose concentration less than or equal to 0.01 mM. Similar to JNK1 activation, we also observed that low glucose‐induced ASK1 activation, dissociation of GRX and thioredoxin (TRX) from ASK1, dimerization of ASK1, and association of Daxx and TRAF2 with ASK1 significantly occurred at a glucose concentration less than or equal to 0.01 mM. J. Cell. Biochem. 89: 653–662, 2003. © 2003 Wiley‐Liss, Inc.
Cell cycle‐dependent transduction of cell‐permeant Cre recombinase proteins
Authors: Jo Daewoong , Lin Qing , Nashabi Abudi , Mays Deborah J. , Unutmaz Derya , Pietenpol Jennifer A. , Ruley H. Earl
Published: July 2003

Protein transduction has been widely used to analyze biochemical processes in living cells quantitatively and under non‐steady‐state conditions. The present study analyzed the effects of cell cycle on the uptake and activity of cell‐permeant Cre recombinase proteins. Previous studies had suggested that the efficiency of recombination and/or protein transduction varied among individual cells, even within a clonal population. We report here that cells in the G1 phase of the cell cycle undergo recombination at a lower rate than cells at other phases of the cell cycle, and that this variation results largely from differences in protein uptake, associated with differences in cell size. These results have implications regarding the mechanism of protein transduction and identify a source of heterogeneity that can influence the response of individual cells to cell‐permeant proteins. J. Cell. Biochem. 89: 674–687, 2003. © 2003 Wiley‐Liss, Inc.
Suppressive effect of endogenous regucalcin on nitric oxide synthase activity in cloned rat hepatoma H4‐II‐E cells overexpressing regucalcin
Authors: Izumi Takako , Tsurusaki Yoshinori , Yamaguchi Masayoshi
Published: July 2003

The role of endogenous regucalcin, which is a regulatory protein in calcium signaling, in the regulation of nitric oxide (NO) synthase activity in the cloned rat hepatoma H4‐II‐E cells was investigated. Hepatoma cells were cultured for 24–72 h in the presence of fetal bovine serum (FBS; 10%). NO synthase activity in the 5,500 g supernatant of cell homogenate was significantly increased by the addition of calcium chloride (10 μM) and calmodulin (2.5 μg/ml) in the enzyme reaction mixture. The presence of trifluoperazine (TFP; 50 μM), an antagonist of calmodulin, inhibited the effect of calcium (10 μM) addition in increasing NO synthase activity, indicating the existence of Ca2+/calmodulin‐dependent NO synthase in hepatoma cells. NO synthase activity was significantly decreased by the addition of regucalcin (10−8 or 10−7 M) in the reaction mixture without or with Ca2+/calmodulin addition. The effect of regucalcin (10−7 M) in decreasing NO synthase activity was also seen in the presence of TFP (50 μM) or EGTA (1 mM). The presence of anti‐regucalcin monoclonal antibody (10–50 ng/ml) in the reaction mixture caused a significant elevation of NO synthase activity. NO synthase activity was significantly suppressed in the hepatoma cells (transfectants) overexpressing regucalcin. This decrease was completely abolished in the presence of anti‐regucalcin monoclonal antibody (50 ng/ml) in the reaction mixture. Moreover, the effect of Ca2+/calmodulin addition in increasing NO synthase activity in the hepatoma cells (wild‐type) was completely prevented in transfectants. The present study demonstrates that endogenous regucalcin has a suppressive effect on NO synthase activity in the cloned rat hepatoma H4‐II‐E cells. J. Cell. Biochem. 89: 800–807, 2003. © 2003 Wiley‐Liss, Inc.
Nuclear distribution of Oct‐4 transcription factor in transcriptionally active and inactive mouse oocytes and its relation to RNA polymerase II and splicing factors
Authors: Parfenov Vladimir N. , Pochukalina Galina N. , Davis Donna S. , Reinbold Rolland , Schöler Hans R. , Murti Kuruganti G.
Published: July 2003

The intranuclear distribution of the transcription factor Oct‐4, which is specifically expressed in totipotent mice stem and germ line cells, was studied in mouse oocytes using immunogold labeling/electron microscopy and immunofluorescence/confocal laser scanning microcopy. The localization of Oct‐4 was studied in transcriptionally active (uni/bilaminar follicles) and inactive (antral follicles) oocytes. Additionally, the Oct‐4 distribution was examined relative to that of the unphosphorylated form of RNA polymerase II (Pol II) and splicing factor (SC 35) in the intranuclear entities such as perichromatin fibrils (PFs), perichromatin granules (PGs), interchromatin granule clusters (IGCs), Cajal bodies (CBs), and nucleolus‐like bodies (NLBs). It was shown that: (i) Oct‐4 is localized in PFs, IGCs, and in the dense fibrillar component (DFC) of the nucleolus at the transcriptionally active stage of the oocyte nucleus; (ii) Oct‐4 present in PFs and IGCs colocalizes with Pol II and SC 35 at the transcriptionally active stage; (iii) Oct‐4 accumulates in NLBs, CBs, and PGs at the inert stage of the oocyte. The results confirm the previous suggestion that PFs represent the major nucleoplasmic structural domain involved in active pre‐mRNA transcription/processing. The colocalization of Oct‐4 with Pol II in both IGCs and PFs in active oocytes (uni/bilaminar follicles) suggests that Oct‐4 is intimately associated with the Pol II holoenzyme before and during transcription. The colocalization of Oct‐4, Pol II, and SC 35 with coilin‐containing structures such as NLBs and CBs at the inert stage (antral follicles) suggests that the latter may represent storage sites for the transcription/splicing machinery during the decline of transcription. J. Cell. Biochem. 89: 720–732, 2003. © 2003 Wiley‐Liss, Inc.
Altered gene expression profile in the kidney of vitamin D receptor knockout mice
Authors: Li Xinmin , Zheng Wei , Li Yan Chun
Published: July 2003

The kidney is a primary target organ of the vitamin D endocrine system, and both vitamin D‐deficiency and vitamin D receptor (VDR) ablation lead to impaired renal functions. As an initial step to understand the molecular basis underlying the renal dysfunctions resulted from VDR inactivation, we used DNA microarray technology to search for changes in the gene expression profile in the kidney of VDR knockout mice. Three independent DNA microarray experiments were performed using Affymetrix GeneChips, which included two replicate comparisons between VDR null and wild‐type littermates, and a third comparison between 1,25‐dihydroxyvitamin D3‐treated and vehicle‐treated wild‐type mice. Based on the assumption that VDR inactivation and vitamin D stimulation cause opposite changes in the expression of vitamin D target genes, we identified 95 genes that displayed the same changes in the two VDR‐null/wild‐type comparisons but an opposite change in the third assay, of which 28 genes were up‐regulated and 67 were down‐regulated in VDR null mice. These genes can be divided into several functional categories involved in vitamin D and steroid metabolism, calcium metabolism and signaling, volume and electrolyte homeostasis, signal transduction, transcriptional regulation, cell adhesion, metabolism, immune response, and other functions. These data provide a basis for further investigations into the molecular bases underlying the physiological abnormalities associated with VDR‐ and vitamin D‐deficiency. J. Cell. Biochem. 89: 709–719, 2003. © 2003 Wiley‐Liss, Inc.
Mammalian transcription factor LSF is a target of ERK signaling
Authors: Pagon Zrinka , Volker Janet , Cooper Geoffrey M. , Hansen Ulla
Published: July 2003

LSF is a mammalian transcription factor that is rapidly and quantitatively phosphorylated upon growth induction of resting, peripheral human T cells, as assayed by a reduction in its electrophoretic mobility. The DNA‐binding activity of LSF in primary T cells is greatly increased after this phosphorylation event (Volker et al. [1997]: Genes Dev 11:1435–1446). We demonstrate here that LSF is also rapidly and quantitatively phosphorylated upon growth induction in NIH 3T3 cells, although its DNA‐binding activity is not significantly altered. Three lines of experimentation established that ERK is responsible for phosphorylating LSF upon growth induction in both cell types. First, phosphorylation of LSF by ERK is sufficient to cause the reduced electrophoretic mobility of LSF. Second, the amount of ERK activity correlates with the extent of LSF phosphorylation in both primary human T cells and NIH 3T3 cells. Finally, specific inhibitors of the Ras/Raf/MEK/ERK pathway inhibit LSF modification in vivo. This phosphorylation by ERK is not sufficient for activation of LSF DNA–binding activity, as evidenced both in vitro and in mouse fibroblasts. Nonetheless, activation of ERK is a prerequisite for the substantial increase in LSF DNA–binding activity upon activation of resting T cells, indicating that ERK phosphorylation is necessary but not sufficient for activation of LSF in this cell type. J. Cell. Biochem. 89: 733–746, 2003. © 2003 Wiley‐Liss, Inc.
Increased expression of disintegrin‐metalloproteinases ADAM‐15 and ADAM‐9 following upregulation of integrins α5β1 and αvβ3 in atherosclerosis
Authors: Al‐Fakhri Nadia , Wilhelm Jochen , Hahn Meinhard , Heidt Martin , Hehrlein Friedrich W. , Endisch Andrea M. , Hupp Thomas , Cherian Sanjay M. , Bobryshev Yuri V. , Lord Reginald S.A. , Katz Norbert
Published: July 2003

Regulation of αvβ3 and α5β1 integrin function plays a crucial role in atherosclerosis. Possible regulators of integrin–matrix interactions are integrin‐binding ADAMs (proteins with a disintegrin‐ and metalloproteinase‐domain), like ADAM‐15 and ADAM‐9. Molecular interactions between ADAM‐15, α5β1, and αvβ3 have been demonstrated. ADAM‐9 and ADAM‐15 were found to be interdependently regulated. This study, therefore, investigated whether the upregulation of integrins α5β1 and αvβ3 was correlated with the expression of integrin‐binding ADAMs in atherosclerotic processes. Human arterial and venous vascular smooth muscle cells (VSMCs) were incubated with PDGF over different time intervals up to a 3‐day culture period. mRNA concentrations, quantified by real‐time RT‐PCR and normalized to PBGD, of integrins αvβ3 and α5β1 were strongly increased after a 12‐h PDGF‐incubation in arterial and venous VSMC. ADAM‐15 and ADAM‐9 mRNA production showed a corresponding increase following integrin upregulation after a 24‐h incubation period. Western blot anaylsis revealed an increased protein expression of integrins and ADAMs in PDGF‐stimulated VSMC. Additionally, mRNA concentrations of atherosclerotic and normal human specimens were quantified by real‐time RT‐PCR. mRNA of ADAMs and integrins was significantly increased in atherosclerotic arteries compared to normal arteries. Immunohistochemistry of these specimens showed an increased expression and codistribution of both ADAMs and integrins in atherosclerosis. In conclusion, upregulation of ADAM‐15 and ADAM‐9 in atherosclerosis appears to follow an increase in α5β1 and αvβ3 integrins. Since α5β1 and αvβ3 are known to promote smooth muscle cell migration and proliferation, upregulation of ADAM‐15 and ADAM‐9 could balance integrin–matrix interactions and cell migration, thus modulating neointima progression. J. Cell. Biochem. 89: 808–823, 2003. © 2003 Wiley‐Liss, Inc.
Mechanisms of free‐radical induction in relation to fenretinide‐induced apoptosis of neuroblastoma
Authors: Lovat Penny E. , Ranalli Marco , Corazzari Marco , Raffaghello Lizzia , Pearson Andy D.J. , Ponzoni Mirco , Piacentini Mauro , Melino Gerry , Redfern Christopher P.F.
Published: July 2003

The mechanisms of fenretinide‐induced cell death of neuroblastoma cells are complex, involving signaling pathways mediated by free radicals or reactive oxygen species (ROS). The aim of this study was to identify mechanisms generating ROS and apoptosis of neuroblastoma cells in response to fenretinide. Fenretinide‐induced ROS or apoptosis of SH‐SY5Y or HTLA 230 neuroblastoma cells were not blocked by Nitro l‐argenine methyl ester (l‐NAME), an inhibitor of nitric oxide synthase. Flavoprotein‐dependent superoxide‐producing enzymes such as NADPH oxidase were also not involved in fenretinide‐induced apoptosis or ROS generation. Similarly, ketoconazole, a cytochrome P450 inhibitor, and inhibitors of cyclooxygenase (COX) were also ineffective. In contrast, inhibition of phospholipase A2 or lipoxygenases (LOX) blocked the induction of ROS and apoptosis in response to fenretinide. Using specific inhibitors of LOX, blocking 12‐LOX but not 5‐ or 15‐LOX inhibited both fenretinide‐induced ROS and apoptosis. The effects of eicosatriynoic acid, a specific 12‐LOX inhibitor, were reversed by the addition of the 12‐LOX products, 12 (S)‐hydroperoxyeicosatetraenoic acid and 12 (S)‐hydroxyeicosatetraenoic acid. The targeting of 12‐LOX in neuroblastoma cells may thus be a novel pathway for the development of drugs inducing apoptosis of neuroblastoma with improved tumor specificity. J. Cell. Biochem. 89: 698–708, 2003. © 2003 Wiley‐Liss, Inc.
Active poly(ADPribose) metabolism in DNAase‐ and salt‐resistant rat testis chromatin with high transcriptional activity/competence
Authors: Mennella Maria Rosaria Faraone , Roma Guglielmo , Farina Benedetta
Published: July 2003

A chromatin fraction, named pP fraction, was prepared from rat testis nuclei, which had been digested with nuclease in order to separate soluble and insoluble chromatin. This fraction resembled nuclear matrix as it was highly resistant to DNAase digestion, had a high content of proteins compared to the low DNA percentage, and a noticeable transcriptional activity. Moreover, poly(ADPribosyl)ation system (i.e., poly(ADPR)polymerase, poly(ADPribose), and acceptor proteins) was still present at high levels. In order to study whether it might be identified as the protein support surrounding chromatin loops, this pP fraction was further analyzed after 3 M NaCl extraction. The 3 M NaCl extract and the highly insoluble pellet, named Nuclear Matrix Pellet, were characterized as it regards DNA, newly synthesized RNA and proteins. Furthermore, poly(ADPribose) metabolism was analyzed by measuring both poly(ADPribose) polymerase and poly(ADPribose) glycohydrolase activities, poly(ADPribose) distribution and by identifying protein acceptors. The final pellet had features of nuclear matrix containing less than 10% DNA and high percentage of proteins; 28% of newly synthesized RNA was still associated with this fraction. Long and branched polyADPribose were found in the nuclear matrix‐like pellet, although ADPribose acceptors (mainly H1 and core histones) appeared to be modified mostly with short ADPribose oligomers. Longest and branched polymers were retained on the top of protein gel, likely bound to automodified poly(ADPribose) polymerase. J. Cell. Biochem. 89: 688–697, 2003. © 2003 Wiley‐Liss, Inc.
Retinoic acid inhibits chondrogenesis of mesenchymal cells by sustaining expression of N‐cadherin and its associated proteins
Authors: Cho Seo‐Hyun , Oh Chun‐Do , Kim Song‐Ja , Kim Il‐Chul , Chun Jang‐Soo
Published: July 2003

Retinoic acid (RA) is a well‐known regulator of chondrocyte phenotype. RA inhibits chondrogenic differentiation of mesenchymal cells and also causes loss of differentiated chondrocyte phenotype. The present study investigated the mechanisms underlying RA regulation of chondrogenesis. RA treatment in chondrifying mesenchymal cells did not affect precartilage condensation, but blocked progression from precartilage condensation to cartilage nodule formation. This inhibitory effect of RA was independent of protein kinase C and extracellular signal‐regulated protein kinase, which are positive and negative regulators of cartilage nodule formation, respectively. The progression from precartilage condensation to cartilage nodule requires downregulation of N‐cadherin expression. However, RA treatment caused sustained expression of N‐cadherin and its associated proteins including α‐ and β‐catenin suggesting that modulation of expression of these molecules is associated with RA‐induced inhibition of chondrogenesis. This hypothesis was supported by the observation that disruption of the actin cytoskeleton by cytochalasin D (CD) blocks RA‐induced sustained expression of cell adhesion molecules and overcomes RA‐induced inhibition of chondrogenesis. Taken together, our results suggest RA inhibits chondrogenesis by stabilizing cell‐to‐cell interactions at the post‐precartilage condensation stage. J. Cell. Biochem. 89: 837–847, 2003. © 2003 Wiley‐Liss, Inc.
Nuclear localization of the Hermes transposase depends on basic amino acid residues at the N‐terminus of the protein
Authors: Michel K. , Atkinson P.W.
Published: July 2003

For the Hermes transposable element to be mobilized in its eukaryotic host, the transposase, encoded by the element, must make contact with its DNA. After synthesis in the cytoplasm, the transposase has to be actively imported into the nucleus because its size of 70.1 kDa prevents passive diffusion through the nuclear pore. Studies in vitro using transient expression of a Hermes–EGFP fusion protein in Drosophila melanogaster Schneider 2 cells showed the transposase was located predominantly in the nucleus. In silico sequence analysis, however, did not reveal any nuclear localization signal (NLS). To identify the sequence(s) responsible for localization of Hermes transposase in the nucleus, truncated or mutated forms of the transposase were examined for their influence on sub‐cellular localization of marker proteins fused to the transposase. Using the same expression system and a GFP–GUS fusion double marker, residues 1–110 were recognized as sufficient, and residues 1–32 as necessary, for nuclear localization. Amino acid K25 greatly facilitated nuclear localization, indicating that at least this basic amino acid plays a significant role in this process. This sequence overlaps the proposed DNA binding region of the Hermes transposase and is not necessarily conserved in all members of the hAT transposable element family. J. Cell. Biochem. 89: 778–790, 2003. © 2003 Wiley‐Liss, Inc.
MPA increases idarubicin‐induced apoptosis in chronic lymphatic leukaemia cells via caspase‐3
Authors: Florio Salvatore , Crispino Luca , Ciarcia Roberto , Vacca Giovanni , Pagnini Ugo , Matteis Andrea de , Pacilio Carmen , D'Andrilli Giuseppina , Kumar Christine , Giordano Antonio
Published: July 2003

The caspase family of protease is speculated to have a crucial role in apoptosis. The effect of treatment with Idarubicin (IDA) and Medroxyprogesterone acetate (MPA), used alone or in combination, on the activation of Caspase‐3 in canine Chronic Lymphatic Leukaemia (CLL) cells was investigated, in order to clarify the mechanism of chemo‐ and hormone‐therapy mediated apoptosis. Caspase activity was determined by a quantitative fluorimetric assay. Apoptosis was monitored by propidium iodide (PI) and nucleosomes assay. Treatment of CLL cells for 24 h with MPA 5 μM did not significantly activate caspase‐3 but its activity was increased almost 5‐fold more with IDA 1 μM (P < 0.05) than control. Treatment of CLL cells with IDA 1 μM in equimolecular association with MPA was able to increase the activation of caspase‐3 induced by IDA of the 61.2% (P < 0.05) in comparison with IDA alone. The activation of caspase‐3 was confirmed evaluating apoptosis by PI and nucleosomes assay. Furthermore, both caspase‐3 activation and apoptosis triggered by IDA alone or in combination with MPA were significantly inhibited by specific caspase‐3 inhibitor AC‐DEVD‐CMK. These findings provide an explanation for IDA and MPA induced‐apoptosis mechanism. J. Cell. Biochem. 89: 747–754, 2003. © 2003 Wiley‐Liss, Inc.
Overexpression of interleukin‐2 receptor α in a human squamous cell carcinoma of the head and neck cell line is associated with increased proliferation, drug resistance, and transforming ability
Authors: Kuhn Deborah J. , Smith David M. , Pross Seth , Whiteside Theresa L. , Dou Q. Ping
Published: July 2003

It has been previously demonstrated that human carcinomas express interleukin‐2 receptor (IL‐2R) α, β, and γ chains. The β and γ chains of IL‐2R have intermediate binding affinity for IL‐2 and are responsible for the intracellular signaling cascades after IL‐2 stimulation. IL‐2Rα lacks the cytoplasmic domain, but is essential for increasing the IL‐2‐binding affinity of other receptors. Overexpression of IL‐2Rα in tumor cells is associated with tumor progression and a poor patient prognosis. To define molecular mechanisms responsible for the effects associated with IL‐2Rα expression, ex vivo experiments were performed with the squamous cell carcinoma head‐and‐neck cancer line, PCI‐13, which was genetically engineered to overexpress the IL‐2Rα chain. While IL‐2Rα‐overexpressing PCI‐13 cells were capable of forming colonies in soft agar, PCI‐13 cells transfected with the control vector or those expressing IL‐2Rγ did not. Consistently, IL‐2Rα‐expressing tumor cells proliferated more rapidly than the control or IL‐2Rγ+ cells, associated with increased levels of cyclins A and D1 and cyclin‐dependent kinase (cdk(s)) 2 and 4 proteins. In addition, IL‐2Rα‐expressing cells were significantly more resistant to apoptosis induction by a tripeptidyl proteasome inhibitor (ALLN) and two chemotherapeutic drugs (VP‐16 and taxol) than the control or IL‐2Rγ+ cells. Accompanying the drug resistance, high levels of anti‐apoptotic Bcl‐XL and Bcl‐2 proteins were found in the mitochondria‐containing fraction of IL‐2Rα‐expressing tumor cells. Treatment of IL‐2Rα‐expressing cells with a specific Janus kinase 3 (Jak3) inhibitor decreased expression of cyclin A, cyclin D1, Bcl‐XL, and Bcl‐2 proteins. Finally, high levels of ubiquitinated proteins were detected in the proliferating IL‐2Rα‐expressing cells. Our data suggest that increased proliferation rates and decreased drug sensitivity of IL‐2Rα‐expressing tumor cells are responsible for the enhanced tumor aggressiveness and poor clinical prognosis of patients whose tumors express IL‐2Rα. J. Cell. Biochem. 89: 824–836, 2003. © 2003 Wiley‐Liss, Inc.
Estrogenic endocrine disruptive components interfere with calcium handling and differentiation of human trophoblast cells
Authors: Derfoul A. , Lin F.J. , Awumey E.M. , Kolodzeski T. , Hall D.J. , Tuan R.S.
Published: July 2003

During development, calcium (Ca) is actively transported by placental trophoblasts to meet fetal nutritional and the skeletal mineralization needs. Maternal exposure to estrogenic pesticides, such as 1,1‐bis(p‐chlorophenyl)‐2,2,2‐trichloroethane (DDT) and methoxychlor (MTC), has been shown to result in reproductive disorders and/or abnormal fetal development. In this study, we have examined the effects of exposure of trophoblastic cells to MTC and DTT, in comparison to 17β‐estradiol (E2) and diethylstilbestrol (DES), to test the hypothesis that cellular Ca handling is a target for these endocrine disruptive components. Treatment with DDT, MTC, DES, or E2 increased cellular Ca uptake, and the expression of trophoblast‐specific human Ca binding protein (HCaBP) was down‐regulated by both MTC and DDT. Treatment with MTC, DDT, and DES inhibited cell proliferation, induced apoptosis, and suppressed expression of several trophoblast differentiation marker genes. These effects were reversed by overexpression of metallothionein IIa, a gene highly responsive to cadmium and other metals. These results strongly suggest that trophoblast Ca handling functions are endocrinally modulated, and that their alteration by candidate endocrine disruptors, such as MTC and DDT, constitutes a possible pathway of the harmful effects of these components on fetal development. J. Cell. Biochem. 89: 755–770, 2003. © 2003 Wiley‐Liss, Inc.
Monoclonal antibody to fibulin‐1 generated by genetic immunization
Authors: Pupa S.M. , Forti S. , Invernizzi A.M. , Giovanazzi R. , Twal W.O. , Argraves W.S. , Ménard Sylvie
Published: July 2003

Fibulin‐1 (Fbln‐1) is an extracellular matrix (ECM) and plasma glycoprotein. Considering the growing evidence indicating that Fbln‐1 plays a role in cancer we sought to develop monospecific antibodies to better facilitate further studies of the function of Fbln‐1 in breast cancer. Using a plasmid expression vector encoding full‐length human Fbln‐1D as an immunogen and CpG oligodeoxyribonucleotides as adjuvant a monoclonal antibody (MAb) against Fbln‐1 was produced. This MAb, designated MEM‐2 was of IgM isotype and reacted with bacterially expressed Fbln‐1. Furthermore, MEM‐2 reacted with Fbln‐1 expressed in the ECM released by cultured human breast carcinoma SKBR‐3 cells in ELISA, and also with Fbln‐1 present in SKBR‐3 cell extract in immunoprecipitation and Western blotting. MEM‐2 also reacted with Fbln‐1 in human breast carcinoma specimens. These findings illustrate the utility of genetic immunization as a means of generating monoclonal antibodies to tumor‐related ECM proteins. MEM‐2 represents a useful new tool for the study of Fbln‐1 in breast cancer. J. Cell. Biochem. 89: 647–652, 2003. © 2003 Wiley‐Liss, Inc.
Vitamin D3 supports osteoclastogenesis via functional vitamin D response element of human RANKL gene promoter
Authors: Kitazawa Sohei , Kajimoto Kazuyoshi , Kondo Takeshi , Kitazawa Riko
Published: July 2003

Receptor activator of NF‐κB ligand (RANKL) has been identified as requisite for osteoclastogenesis. To elucidate the molecular mechanism that conducts its catabolic action on bone, the effect of 1α,25 dihydroxyvitamin D3 (1α,25(OH)2D3) on osteoclastogenesis and RANKL mRNA expression was examined by coculture, RT‐PCR and nuclear run‐on studies. By accelerating the transcription rate of the RANKL gene in SaOS2 osteoblastic cells, 1α,25(OH)2D3 enhanced in vitro osteoclast formation from peripheral monocytes. Cloning and characterization of the 5′‐flanking region of the human RANKL gene revealed that the basic promoter comprises inverted TATA‐ and CAAT‐boxes flanked by RUNX2 binding sites. Both electrophoresis mobility shift assay (EMSA) and transfection studies demonstrated that 1α,25(OH)2D3 activated human RANKL promoter through vitamin D responsive elements (VDRE) located at −1584/−1570 by binding VDR and RXRα heterodimers in a ligand‐dependent manner. The results provide direct evidence that 1α,25(OH)2D3 augments osteoclastogenesis by transactivating the human RANKL gene in osteoblastic cells through VDRE. J. Cell. Biochem. 89: 771–777, 2003. © 2003 Wiley‐Liss, Inc.
Effect of heat treatment and evaporation on patulin and some other properties of apple juice
Authors: Kadakal Çetin , Nas Sebahattin
Published: July 2003

Patulin, hydroxymethylfurfural (HMF), colour and clarity values of 20 apple juice samples with different Brix contents were determined after heat treatment (90 and 100 °C for 5, 10, 15 and 20 min under atmospheric pressure) and evaporation (70 and 80 °C for 5, 10, 15 and 20 min) processes. As the heating and evaporation times increased, the concentration of patulin in the apple juice samples decreased. The 90 and 100 °C heat treatments resulted in reductions in patulin concentration of 18.81 and 25.99% respectively after 20 min; the corresponding values were 9.40 and 14.06% for 70 and 80 °C evaporation respectively. The increase in HMF was higher with heat treatment than with evaporation. Evaporation decreased the colour values of the samples, whereas heat treatment increased them.

© 2003 Society of Chemical Industry
Comparative study on proteolysis of two species of bigeye snapper, Priacanthus macracanthus and Priacanthus tayenus
Authors: Benjakul Soottawat , Leelapongwattana Kittima , Visessanguan Wonnop
Published: July 2003

Proteolytic activity in muscle from two species of bigeye snapper (Priacanthus macracanthus and Priacanthus tayenus) was studied. Autolysis of mince and washed mince at 50 and 60 °C was compared. Higher degradation of myosin heavy chain was observed in both mince and washed mince from P macracanthus than in those from P tayenus, especially when the incubation time was increased. Autolysis of washed mince from both species was inhibited by soybean trypsin inhibitor, suggesting that myofibril‐associated proteases were serine proteases. When sarcoplasmic proteolytic activity in P macracanthus muscle was studied, two activity peaks with an optimum temperature of 60 °C were observed at pH 6.5 and 8.5. The activities of both peaks were mostly inhibited by soybean trypsin inhibitor, suggesting that the major protease was a serine protease. Major sarcoplasmic proteolytic activity in P macracanthus muscle was found at Mr 62 000 on sodium dodecyl sulphate substrate gel. For P tayenus sarcoplasmic proteolytic activity, two activity peaks with an optimum temperature of 60 °C were found at pH 5.0 and 8.5. The pH 5.0 peak activity was effectively inhibited by pepstatin A, while the pH 8.5 peak activity was inhibited by several inhibitors. The results indicated that various sarcoplasmic proteases were present in P tayenus muscle. The two species contained different sarcoplasmic proteases in terms of composition and activity level. P macracanthus muscle generally had higher sarcoplasmic proteolytic activities than P tayenus muscle. Copyright © 2003 Society of Chemical Industry
Macromolecular characteristics of ten yam (Dioscorea spp) starches
Authors: Rolland‐Sabaté Agnès , Georges Amani N'Guessan , Dufour Dominique , Guilois Sophie , Colonna Paul
Published: July 2003

Starches from ten yam (Dioscorea) species were compared with those of maize, wheat, potato and cassava, and characterized by high‐performance size‐exclusion chromatography coupled with multiangle laser light scattering. Treatment with 95% (v/v) dimethylsulphoxide and microwave heating in a high‐pressure vessel led to complete dissolution of the starch samples. For yam starches, M̄w were between 1.88 × 108 and 3.27 × 108 g mol−1 and R̄G were between 258 and 396 nm. The hydrodynamic coefficients of amylopectins were between 0.36 and 0.44, indicating that those of maizes and esculenta 5 and dumetorum yam species had particularly highly branched structures. Multidimensional analysis of the macromolecular characteristics of yam starches indicated three classes: dumetorum cultivar (Dioscorea dumetorum), esculenta 5 cultivar (Dioscorea esculenta) and the other eight yam starches, including cultivars of Dioscorea alata and Dioscorea cayenensis‐rotundata species. Some yam starches were also leached at 90 °C. The macromolecular characteristics of the leached fractions confirmed the previous typology. Copyright © 2003 Society of Chemical Industry
Enzymatic hydrolysis of Nile tilapia (Oreochromus niloticus) myofibrillar proteins: effects on nutritional and hydrophilic properties
Authors: Cândido Lys Mary Bileski , Sgarbieri Valdemiro Carlos
Published: July 2003

Protein concentrate (PC) was prepared from eviscerated and mechanically deboned fish. The pulp was washed sequentially with 0.05 M NaCl and 4 g l−1 NaHCO3 solutions, with cold ethanol and then with hot (50 °C) ethanol. Subsequently, it was dried (40 °C) and ground to 60 mesh. Enzyme (Flavourzime®) hydrolysates were prepared with a degree of hydrolysis (DH) ranging from 3.5 to 45%. The PC was characterized for proximate composition, amino acid profile, physical and functional properties. The influence of DH on functional and nutritional properties was evaluated. Hygroscopicity increased in all hydrolysates for environmental relative humidities above 40% and solubility increased rapidly for DH above 7%. Water retention, water absorption and oil absorption all decreased as a function of DH. The pH (3 to 9) influenced the water and oil absorption of the hydrolysate with 7% DH. A pH near the isoelectric point tends to decrease water absorption but increase oil absorption capacity. The insoluble fraction (IF) of the hydrolysates presented a better amino acid profile than the soluble fraction (SF). The nutritional indexes determined for the IF did not differ from the total hydrolysate, but they were consistently higher than those for the SF. Copyright © 2003 Society of Chemical Industry
Comparative aroma profiles using supercritical carbon dioxide and Likens–Nickerson extraction from a commercial brand of Basmati rice
Authors: Bhattacharjee Paramita , Ranganathan TV , Singhal Rekha S , Kulkarni Pushpa R
Published: July 2003

A comparative evaluation of the extraction of the aroma constituents of a popular commercial brand of Basmati rice using Likens–Nickerson extraction and supercritical fluid extraction (SFE) with carbon dioxide was carried out. SFE at 50 °C and 120 bar for 2 h provided appreciable extraction of the volatile constituents of the rice as compared with Likens–Nickerson extraction. The advantages of smaller sample size, shorter time of extraction and negligible possibility of artefacts with the SFE technique merit its use for recovery of aroma volatiles from Basmati rice. Copyright © 2003 Society of Chemical Industry
Crystallisation and moisture sorption properties of selected Australian unifloral honeys
Authors: Yao Lihu , Bhandari Bhesh R , Datta Nivedita , Singanusong Riantong , D'Arcy Bruce R
Published: July 2003

The sorption properties of yapunyah (Eucalyptus ochropholia) and yellow box (Eucalyptus melliodora) honeys (Australian unifloral honeys) were investigated in a controlled relative humidity (RH) environment at 30 °C for 71 days. The original water activity of the honeys affected the sorption properties. These two honeys absorbed moisture at and above 67.9% RH and desorbed moisture at and below 51.4% RH. The crystallisation behaviour of tea tree (Melaleuca quinquenervia) and yapunyah honeys was studied during storage at 13 and 23 °C. The degree of crystallisation was monitored by measuring the absorbance at 660 and 665 nm using a spectrophotometer. The heat‐treated honeys did not show any sign of crystallisation after 5 months, whereas a seeding with precrystallised honey induced crystallisation of the same honeys. This crystallisation was more rapid at 13 than at 23 °C. Copyright © 2003 Society of Chemical Industry
Functional and multiple end‐use characterisation of Canadian wheat using a reconstituted dough system
Authors: Uthayakumaran Surjani , Lukow Odean M
Published: July 2003

Three Canadian wheat cultivars representing the Canada Western Red Spring, Canada Western Extra Strong and Canada Prairie Spring classes, varying in protein content yet containing similar high‐molecular‐weight glutenin subunits, were evaluated for dough functionality and multiple end‐use properties. The effect of protein content on dough properties and end‐product quality was also studied. Gluten, starch and water‐soluble components were extracted from the flours and reconstituted to make up three samples for each variety to match the protein content of the three parent flours. Empirical and dynamic dough rheological properties, baking (bread and tortilla) performance and noodle‐making properties of the flours were determined using small‐scale techniques. Results indicated that protein content had a significant effect on rheological and end‐use quality of wheat flours. Increase in protein content (of the reconstituted dough) increased mixograph peak height (r = 0.761), peak width, maximum resistance to extension and end‐product quality characteristics such as loaf volume (r = 0.906), noodle firmness and cutting force and decreased storage and loss moduli. Reconstituted flours from the three varieties at the same protein content also showed significant differences in mixing time, mixograph peak height, maximum resistance to extension, composite fineness of pan bread, tortilla diameter, cooked noodle hardness, gumminess and dynamic viscosity of dough. This study indicates that a simple reconstituted dough system can provide an unambiguous assignment of the quantitative and qualitative effects of dough components on dough properties. It has the advantages that any aspect of flour composition can be manipulated and details of the relation between composition and functional behaviour can be obtained for any end‐product. Copyright © 2003 Crown in the right of Canada. Published by Society of Chemical Industry
Effect of NaCl on textural changes and protein and lipid degradation during the ripening stage of sufu, a Chinese fermented soybean food
Authors: Han Bei‐Zhong , Wang Jia‐Huai , Rombouts Frans M , Nout MJ Robert
Published: July 2003

Sufu is made by solid state fungal fermentation (using Actinomucor elegans) of tofu, followed by salting and maturation in dressing mixtures containing salt, alcohol and various other ingredients. NaCl in dressing mixtures strongly affected the changes in textural properties and the hydrolysis of protein and lipid of sufu. Higher salt contents (14% w/w) resulted in increased hardness (+100%) and elasticity (+18%) and reduced adhesiveness (−30%). Hardness and elasticity could be used to judge the extent of sufu ripening. SDS‐PAGE showed the disappearance of all protein subunits at 80 and 110 g kg−1 salt content; however, some protein subunits were still detectable at 140 g kg−1 salt content after 60 days of ripening. Higher ratios of free amino nitrogen to total nitrogen (FAN/TN = 0.4–0.45) and free amino acids to crude protein (FAA/CP = 0.24–0.26) were observed in sufu with lower (80 g kg−1) salt content. FAN/TN and FAA/CP in white sufu (obtained with dressing mixtures containing only salt and alcohol) were higher than those in red sufu (obtained with dressing mixtures containing angkak or kojic red rice) owing to different dressing mixture compositions. Increases in free fatty acids (FFA) were also observed during ripening. FFA levels in sufu with lower salt content increased rapidly during the first 30–40 days and then increased slowly, probably resulting from the formation of fatty acid esters. Lowering the salt content (80 g kg−1) can shorten the ripening time to 40 days, which is of benefit to manufacturers. However, sufu will spoil, ie undergo souring, during the ripening stage at salt contents of 50 g kg−1 or lower. Copyright © 2003 Society of Chemical Industry
Classification of monovarietal Italian olive oils by unsupervised (PCA) and supervised (LDA) chemometrics
Authors: Giansante Lucia , Di Vincenzo Daria , Bianchi Giorgio
Published: July 2003

This research work was conducted to establish whether monovarietal olive oils could be differentiated by their basic classes of compounds, ie (a) fatty acids, (b) fatty alcohols, (c) polycyclic triterpenes and (d) squalene. The ratio values of biosynthetically correlated acids were also examined. The mentioned classes of compounds, formed in distinct biosynthetic compartments of the olive fruit, should represent characteristic compositional data of an olive cultivar. The widely cultivated Italian olive cultivars studied were Frantoio, Bosana, Dritta and Leccino. Principal component analysis (PCA) was applied to the analytical data to reveal the compounds (variables) with the highest weights (loadings), with the aim of using them in subsequent computations. These variables were tetracosanol, hexacosanol Δ5‐avenasterol, cycloartenol, 24‐methylencycloartenol, oleic, linoleic, linolenic, stearic and palmitoleic acids and the ratios palmitic/stearic, palmitic/palmitoleic and linoleic/linolenic. Linear discriminant analysis (LDA), carried out on a training set of 57 oils (13 Dritta, 25 Leccino, 12 Frantoio and seven Bosana) produced a 96% correct group classification. The prediction LDA model created with the training set was validated with a test set of 19 oil samples (six Dritta, seven Leccino, four Frantoio and two Bosana), permitting accurate classification of all the ‘unknown’ olive oils. Copyright © 2003 Society of Chemical Industry
Chickpea leaves as a vegetable green for humans: evaluation of mineral composition
Authors: Ibrikci Hayriye , Knewtson Sharon JB , Grusak Michael A
Published: July 2003

Chickpea (Cicer arietinum) is generally consumed as a seed food, being a good source of protein and other essential human nutrients. However, young chickpea leaves are also eaten as a cooked vegetable green in certain parts of the world and could be a useful source of dietary nutrients, especially in malnourished populations. Because little information is available on the mineral content of this food, we characterised leaf mineral concentrations in 19 diverse accessions of chickpea. Both desi and kabuli chickpea types were studied. All plants were greenhouse‐grown and were fertilised daily with a complete mineral solution. Young, fully expanded leaves (fourth through seventh nodes from the apex) were harvested at both early and late vegetative stages. The leaves were dried, ashed and analysed for mineral concentrations. Macronutrient mineral (Ca, Mg, K, P) concentrations varied from 1.3‐fold to 1.8‐fold and micronutrient mineral (Fe, Zn, Mn, Cu, B, Ni) concentrations varied from 1.5‐fold to 2.4‐fold across all accessions. No major differences were observed in leaf mineral concentrations between the kabuli and desi types; mineral concentrations were generally lower in leaves collected at the later harvest date. Microscopic analyses demonstrated that all accessions contained crystal inclusions, suggestive of calcium oxalate crystals. Overall, chickpea leaves were found to be a good source of several minerals required by humans, and the levels of most of these minerals significantly exceeded those previously reported for spinach and cabbage. Published in 2003 by Society of Chemical Industry[This is a U.S. Government work and is in the public domain in the U.S.A.]
Characteristics of olive fruits and extra‐virgin olive oils obtained from olive trees growing in Appellation of Controlled Origin ‘Sierra Mágina’
Authors: Rial Diego J , Falqué Elena
Published: July 2003

Olives and olive oils from Appellation of Controlled Origin (ACO) ‘Sierra Mágina’ have been analysed during the 1997/98 harvest. The territory of this ACO has been subdivided into four zones of different characteristics. The olives were harvested on two occasions (November 1997 and January 1998). Various parameters were analysed, such as the ripening index, the average volume, the average weight of 100 olives and 100 stones, the pulp/stone ratio, the industrial yield, etc. From these olives, olive oils were extracted using an Abencor system, and the free acidity, the peroxide index, the coefficients K270 and K232 and the fatty acid composition were determined. The analysis of the results obtained, as well as principal component analysis, demonstrates a marked variability in the fruits, but the corresponding olive oils show fairly homogeneous compositions that are difficult to distinguish by sensory analysis. The olive oils from this ACO are extra‐virgin olive oils characterised by low acidity, a high content of oleic acid and a high ratio of unsaturated to saturated fatty acids. The most important sensory attributes are fruity, woody‐fig, green, bitter and pungent. Copyright © 2003 Society of Chemical Industry
Effect of osmotic stress on microstructure and mass transfer in onion and strawberry tissue
Authors: Ferrando Montserrat , Spiess WEL
Published: July 2003

We investigated the structural changes in onion epidermis and strawberry cortex tissue due to osmotic stress during osmotic treatment (OT) with sucrose solutions of concentrations ranging from 300 to 600 g kg−1. We used miniaturised experiments with confocal scanning laser microscopy (CSLM) to monitor microstructural changes—cellular shrinkage and viability—during OT. The overall cellular shrinkage of these plant tissues was not significantly different under the same conditions of osmotic stress. For onion epidermis we determined the transmembrane water flux from the cellular shrinkage and found that the coefficient of water membrane permeability was (1.0 ± 0.39) × 10−6 mol2 J−1 m−2 s−1. Osmotic stress did not affect the cellular viability of onion epidermis but significantly reduced the viability of protoplasts of strawberry in the whole range of solution concentrations. We used Fick's unsteady state second‐order diffusion equation to describe cellular water transport under transient conditions. The pseudo‐diffusional approximation led to water diffusivity values in the range (3–10) × 10−12 m2 s−1 for both onion epidermis and strawberry cortex tissue. The concentration of the solution did not significantly affect the values of effective water diffusivity, which indicated the primary role of the plasma membrane during the initial stages of OT. Copyright © 2003 Society of Chemical Industry