bluuebunny/crossref_metadata_2025 huggingface dataset:
Survivin study: What is the next wave?
Authors: Li Fengzhi
Published: August 2003

Survivin, a novel member of inhibitor of apoptosis (IAP) protein family, is aberrantly expressed in cancer but undetectable in normal, differentiated adult tissues. Current studies suggest that survivin is implicated in both control of apoptosis and regulation of cell division. However, due to some inconsistent observations on survivin subcellular localization, there is debate about survivin's function in regulating apoptosis, cell division, or both. This review will discuss concepts, experimental methods, and interesting results that unify the different notions about survivin localization and function or point out gaps of knowledge about controversial issues. The author also intends to review various aspects of survivin studies, which were not emphasized or sufficiently discussed in previous reviews on survivin, and update recent developments that may reveal new applications of disease‐oriented therapeutics in the coming years. J. Cell. Physiol. 197: 8–29, 2003 © 2003 Wiley‐Liss, Inc.
PKC‐ζ is required for angiotensin II‐induced activation of ERK and synthesis of C‐FOS in MCF‐7 cells
Authors: Muscella Antonella , Greco Simona , Elia Maria Giovanna , Storelli Carlo , Marsigliante Santo
Published: August 2003

We examined the signalling pathways responsible for the Ang II induction of growth in MCF‐7 human breast cancer cells. Ang II in MCF‐7 cells induced: (a) the translocation from the cytosol to membrane and nucleus of atypical protein kinase C‐ζ (PKC‐ζ) but not of PKC‐α, ‐δ, ‐ε and ‐η; (b) the expression of c‐fos mRNA and protein; (c) the phosphorylation of the extracellular signal‐regulated protein kinases 1 and 2 (ERK1/2). All these effects were due to the activation of the Ang II type I receptor (AT1) since they were blocked by the AT1 antagonist losartan. The Ang II‐stimulated ERK1/2 phosphorylation was blocked by (a) high doses of staurosporine, inhibitor of PKC‐ζ, and by a synthetic myristoylated peptide with sequences based on the endogenous PKC‐ζ pseudosubstrate region (ζ‐PS); (b) PD098059, a mitogen‐activated protein kinase kinase inhibitor (MAPKK/MEK); and, moreover, (c) the inhibitors of phosphoinositide 3‐kinases (PI3K), LY294002 and wortmannin, thus indicating that PI3K may act upstream of ERK1/2. The Ang II‐evoked c‐fos induction was blocked only by high doses of staurosporine and by ζ‐PS whilst PD098059, LY294002 and wortmannin were ineffective, thus indicating that c‐fos induction is not due to ERK1/2 activity. When the epidermal growth factor‐receptor (EGFR) tyrosine kinase activity was inhibited by the use of its inhibitor AG1478, Ang II was still able to induce ERK1/2 phosphorylation and c‐fos expression, therefore proving that the transactivation of EGFR was not required for these Ang II effects in MCF‐7 cells. The previously reported proliferation of MCF‐7 cells induced by Ang II was blocked by PD098059 and by wortmannin in a dose‐dependent manner, thereby indicating that in MCF‐7 cells the PI3K and ERK pathways mediate the mitogenic signalling of AT1. Our results suggest that in MCF‐7 cells Ang II activates multiple signalling pathways involving PKC‐ζ, PI3K and MAPK; of these pathways only PKC‐ζ appears responsible for the induction of c‐fos. J. Cell. Physiol. 197: 61–68, 2003© 2003 Wiley‐Liss, Inc.
Kaempferol‐induced growth inhibition and apoptosis in A549 lung cancer cells is mediated by activation of MEK‐MAPK
Authors: Nguyen T.T.T. , Tran E. , Ong C.K. , Lee S.K. , Do P.T. , Huynh T.T. , Nguyen T.H. , Lee J.J. , Tan Y. , Ong C.S. , Huynh H.
Published: August 2003

A vast variety of naturally occurring substances have been shown to protect against experimental carcinogenesis and an increasing amount of evidence suggests that kaempferol may have cancer chemopreventative properties. However, the precise underlying protective mechanisms are poorly understood. To elucidate these mechanisms, we challenged human lung cancer cell line A549 with kaempferol and investigated its effects upon cellular growth and signal transduction pathways. Treatment of A549 cells with kaempferol resulted in a dose‐ and time‐dependent reduction in cell viability and DNA synthesis with the rate of apoptosis equivalent to 0.9 ± 0.5, 5.2 ± 1.5, 16.8 ± 2.0, 25.4 ± 2.6, and 37.8 ± 4.5% on treatment with 0, 17.5, 35.0, 52.5, and 70.0 μM kaempferol, respectively. Concomitantly, kaempferol treatments led to a 1.2‐, 2.7‐, 3.3‐, and 3.4‐fold increase in Bax. Similar elevations were also observed in Bad which increased 1.2‐, 3.3‐, 3.7‐, and 4.7‐fold, respectively, as compared to control. Bcl‐2 and Bcl‐xL expression were inhibited in a dose‐dependent fashion. While the Akt‐1 and phosphorylated Akt‐1 were inhibited, the mitogen‐activated protein kinase (MAPK) was activated upon kaempferol treatment. Kaempferol induced apoptosis was associated with the cleavage of caspase‐7 and poly ADP‐ribose polymerase (PARP). Inhibition of MEK1/2 but not PI‐3 kinase blocked kaempferol‐induced cleavage of caspase‐7, PARP cleavage, and apoptosis. The results suggest that inactivation of Akt‐1 and alteration of Bcl‐2 family of proteins are not sufficient for kaempferol to induce apoptosis and activation of MEK‐MAPK is a requirement for kaempferol‐induced cell death machinery in A549 cells. J. Cell. Physiol. 197: 110–121, 2003© 2003 Wiley‐Liss, Inc.
Downregulation of a rheumatoid arthritis‐related antigen (RA‐A47) by ra‐a47 antisense oligonucleotides induces inflammatory factors in chondrocytes
Authors: Hattori Takako , Kawaki Harumi , Kubota Satoshi , Yutani Yasutaka , Crombrugghe Benoit de , Mark Klaus von der , Takigawa Masaharu
Published: August 2003

Previously we have shown that the expression of RA‐A47 (rheumatoid arthritis‐related antigen) which is identical to HSP47, a collagen‐binding chaperon, is downregulated in chondrocytes by tumor necrosis factor α (TNFα). RA‐A47 was also found on the surface of chondrocytes where it is recognized as an antigen in the serum of rheumatoid arthritis (RA) patients. Its translocation to the cell surface from endoplasmic reticulum membrane where it is normally located was also enhanced by TNFα. To understand the significance of RA‐A47 downregulation in chondrocytes independent from other effects of TNFα, we used an antisense oligonucleotide approach and investigated the effect of this treatment on the expression of molecules related to matrix degradation and production of growth factors for chondrocytic, endothelial, and synovial cells. Here we show that treatment of rabbit chondrocyes and human chondrosarcoma cells HCS‐2/8 by ra‐a47 antisense S‐oligonucleotides significantly reduced the expression of ra‐a47 both at mRNA and protein level. Interestingly, this TNFα‐independent RA‐A47 downregulation was associated with a strong induction of matrix metalloproteinase (MMP)‐9 mRNA and inducible NO synthase (iNOS) mRNA. The induction of active‐type MMP‐9 was further detected by gelatin zymography. Under the same conditions, the release of basic fibroblast growth factor (bFGF) and connective tissue growth factor (CTGF) from HCS‐2/8 cells into the conditioned medium (CM) was strongly enhanced. These effects were not a result of TNFα upregulation, since the ra‐a47 antisense oligonucleotide treatment did not enhance TNFα synthesis. These observations indicate that downregulation of RA‐A47 induces TNFα‐independent cartilage‐degrading pathways involving iNOS and MMP‐9. Furthermore, the stimulation of bFGF and CTGF release from chondrocytes may stimulate the proliferation of adjacent endothelial and/or synovial cells. J. Cell. Physiol. 197: 94–102, 2003© 2003 Wiley‐Liss, Inc.
Phenotypic change regulates monocyte chemoattractant protein‐1 (MCP‐1) gene expression in human retinal pigment epithelial cells
Authors: Uetama Tomoko , Ohno‐Matsui Kyoko , Nakahama Ken‐ichi , Morita Ikuo , Mochizuki Manabu
Published: August 2003

We investigated the expression profile of monocyte chemoattractant protein‐1 (MCP‐1) in human retinal pigment epithelial (RPE) cells under different culture conditions and evaluated the molecular mechanism responsible for MCP‐1 gene expression in RPE cells. After cellular confluence, total RNA was extracted and used for RT‐PCR. Medium conditioned by RPE was used for ELISA and Western blotting. The result showed that RPE cells cultured on plastic expressed MCP‐1 constitutively in the absence of any stimuli. On the other hand, growing human RPE on laminin‐coated flasks instead of plastic reduced the production of MCP‐1. In the RPE cells cultured on plastic, IκB was degraded and A20 protein increased concomitantly. MCP‐1 upregulation in RPE cells on plastic was attenuated by the addition of MG‐132, a proteasome inhibitor. Also, the addition of pyrolidine dithiocarbonate (PDTC) and hypoxic conditions (0.5% O2) decreased MCP‐1 production in these cells. These findings suggested that the expression profile of MCP‐1 is regulated by phenotypic alterations of the RPE cells. And the increased MCP‐1 expression in RPE cells cultured on plastic is caused via spontaneous activation of NFκB induced by susceptibility to oxidative stress. J. Cell. Physiol. 197: 77–85, 2003© 2003 Wiley‐Liss, Inc.
Rat brain pericyte cell lines expressing β2‐adrenergic receptor, angiotensin II receptor type 1A, klotho, and CXCR4 mRNAs despite having endothelial cell markers
Authors: Asashima Tomoko , Iizasa Hisashi , Terasaki Tetsuya , Nakashima Emi
Published: August 2003

Pericytes are an integral component of blood capillaries, but their involvement in a variety of conditions and diseases, including hypertension and multiple sclerosis, is poorly understood. In order to analyze the mRNA expression of markers related to hypertension and multiple sclerosis in rat brain pericytes, we have established brain capillary pericyte cell lines from temperature‐sensitive SV40 large T antigen transgenic rats. The newly established clones showed similar biochemical and morphological properties to primary pericytes. The expression of endothelial cell‐related markers Flt‐1, Flk‐1, Tie‐1, and Tie‐2 was evaluated by RT‐PCR analysis. β2‐Adrenergic receptor (β2‐AR), angiotensin II receptor type1A (AT1A), and klotho were also evaluated as markers related to hypertension and multiple sclerosis. All of the isolated clones expressed β2‐AR, AT1A and klotho genes. They also stably expressed Flt‐1 and Tie‐2, while Flk‐1, Tie‐1 and CXCR4 were expressed only at low levels in some of the clones. The expressions of AT1 in TR‐PCT1 were determined by Western blotting. Angiotensin II stimulated migration of pericytes. This effect was blocked by an AT1 antagonist. The pericyte cell lines established here are pluripotent, and should be useful for analysis of the reactivity and biological roles of pericytes. J. Cell. Physiol. 197: 69–76, 2003© 2003 Wiley‐Liss, Inc.
IGF‐1 enhances a store‐operated Ca2+ channel in skeletal muscle myoblasts: Involvement of a CD20‐like protein
Authors: Ju Yue‐Kun , Wu Ming‐Jie , Chaulet Herve , Marciniec Tadeusz , Graham Robert M. , Allen David G.
Published: August 2003

Overexpression of IGF‐1 in C2C12 myoblasts causes hypertrophy when myoblasts fuse to form myotubes, a response that requires elevated intracellular calcium. We show that myoblasts contain a store‐operated Ca2+ channel (SOCC) whose activity is enhanced with IGF‐1 overexpression. A membrane protein, CD20, can cause Ca2+ entry, which is increased by IGF‐1. We therefore tested whether CD20 mediates the SOCC activity in myoblasts. An antibody to the extracellular loop of CD20 detected a protein in myoblasts and this antibody also inhibited Ca2+ entry through SOCC. Overexpression of CD20 in myoblasts increased SOCC activity. However, we could not detect mRNA for CD20 in myoblasts and an antibody to the intracellular C‐terminus of CD20 was unable to detect CD20 in these cells. These studies demonstrate that CD20 is a novel SOCC or modulates SOCC activity. However, the SOCC activity observed in C2C12 myoblasts is mediated not by CD20, but by a CD20‐like protein. Activation of this SOCC may contribute to IGF‐1‐induced hypertrophy in these cells. J. Cell. Physiol. 197: 53–60, 2003© 2003 Wiley‐Liss, Inc.
Gicerin, an Ig‐superfamily cell adhesion molecule, promotes the invasive and metastatic activities of a mouse fibroblast cell line
Authors: Tsukamoto Yasuhiro , Egawa Michiko , Hiroi Satoshi , Furuya Masaru , Tsuchiya Seiichiro , Sasaki Fumihiko , Miki Naomasa , Taira Eiichi
Published: August 2003

Gicerin is a cell adhesion molecule in the immunoglobulin (Ig) superfamily and plays an important role during development through its adhesive properties. Gicerin has two isoforms that differ in their cytoplasmic domains; s‐gicerin is the shorter and l‐gicerin the longer form of the protein. Gicerin is over‐expressed in some sporadic tumors as well as in developing tissues. To provide direct evidence that gicerin has the potential to participate in malignant aspects of tumor cell behavior, a gicerin cDNA was introduced into L‐929 cells, an endogenous gicerin‐negative mouse fibroblast and subsequently analyzed for changes in their invasive and metastatic potential by implantation into nude mice and chick embryos. Compared with parental cells, both gicerin isoform transfectants showed an enhanced cell growth and invaded deeply into surrounding tissues from implanted sites in both animal models. Furthermore, l‐gicerin transfectants markedly enhanced metastasis to the lung. These findings suggest that gicerin promotes the tumor growth and invasion, and the isoform bearing the longer cytoplasmic domain may play a role in metastasis. J. Cell. Physiol. 197: 103–109, 2003© 2003 Wiley‐Liss, Inc.
Regulation of the pro‐angiogenic microenvironment by carboxyamido‐triazole
Authors: Oliver Vyta Kulpa , Patton Angela M. , Desai Sudhen , Lorang Dominique , Libutti Steven K. , Kohn Elise C.
Published: August 2003

Anti‐angiogenic agents regulate tumor growth by inhibiting endothelial cell proliferation and invasion. Carboxyamido‐triazole (CAI), an inhibitor of non‐voltage‐operated calcium entry and calcium influx‐mediated pathways, has angiogenesis and invasion inhibitory activity. We hypothesized that CAI may express its anti‐angiogenic effects through negative regulation of pro‐angiogenic cytokine production and/or function. In vivo, orally administered CAI prevented A2058 human melanoma xenograft growth and concomitantly resulted in a marked reduction in circulating vascular endothelial growth factor (VEGF) and interleukin‐8 (IL‐8). In vitro, A2058 cell secretion of VEGF was inhibited by CAI treatment under limiting micronutrient conditions that approximate the tumor microenvironment, media restriction, and acidification to pH 6.8 (P = 0.0003 and P = 0.0006, respectively). VEGF and HIF‐1α message and protein were also reduced by CAI treatment. Oral CAI treatment reduced vascular ingrowth in vivo into VEGF‐containing Matrigel plugs. Commensurate with those findings, human umbilical vein endothelial cell (HUVEC) migration towards VEGF was reduced below background by exposure to CAI in the migration chamber (P < 0.0001). An 88% reduction in circulating IL‐8 concentration was measured in CAI‐treated animals. However, IL‐8 protein secretion and gene expression were increased by CAI treatment in culture (P ≤ 0.01), where CAI caused a dose‐dependent acidification of the culture milieu (P ≤ 0.005). This paradox suggests that IL‐8 production in vitro may be more sensitive to ambient pH than cytosolic calcium. These observations suggest that CAI inhibition of tumor cell VEGF production and endothelial cell response to VEGF results in disruption of signaling between the tumor and its microenvironment, causing a net anti‐angiogenic effect. J. Cell. Physiol. 197: 139–148, 2003© 2003 Wiley‐Liss, Inc.
Acute and chronic leptin treatment mediate contrasting effects on signaling, glucose uptake, and GLUT4 translocation in L6‐GLUT4myc myotubes
Authors: Tajmir Panteha , Kwan Jamie Jun‐Mae , Kessas Mona , Mozammel Shehzin , Sweeney Gary
Published: August 2003

We have previously shown that in L6‐GLUT4myc rat skeletal muscle cells, acute treatment with leptin reduced insulin‐stimulated glucose uptake without altering insulin‐stimulated GLUT4 translocation. In contrast, we show here that the ability of leptin to increase phosphorylation of its receptor and to reduce insulin‐stimulated glucose uptake was lost in cells that were continuously exposed to leptin for 24 h. This desensitization correlated with an increase in expression of suppressor of cytokine signaling‐3 (SOCS‐3). Time course analysis demonstrated that the transition from acute to chronic effects of leptin occurs after 2 h. The desensitization of leptin action at 24 h was not reversed by 30 min washout period prior to re‐exposing cells to leptin. However, despite insulin‐stimulated glucose uptake being unaffected upon 24 h preincubation with leptin, a small but significant decrease (37%) in insulin‐stimulated GLUT4 translocation and phosphorylation of Akt on T308 was detected. Insulin‐stimulated phosphorylation of Akt on S473 or of p38 MAPK were unaffected. These results suggest that the chronic leptin treatment leads to desensitization of leptin signaling yet can simultaneously decrease the ability of insulin to phosphorylate Akt on T308 and translocate GLUT4. However, this does not manifest as a reduction in total glucose uptake into L6 myotubes. J. Cell. Physiol. 197: 122–130, 2003© 2003 Wiley‐Liss, Inc.
Urokinase type plasminogen activator receptor is involved in insulin‐like growth factor‐induced migration of rhabdomyosarcoma cells in vitro
Authors: Gallicchio Marisa A. , Kaun Christoph , Wojta Johann , Binder Bernd , Bach Leon A.
Published: August 2003

Urokinase‐type plasminogen activator (uPA) binds to its receptor, uPAR, on the surface of cancer cells, leading to the formation of plasmin. Rhabdomyosarcoma (RMS) cell lines secrete high levels of insulin‐like growth factor II (IGF‐II), suggesting autocrine IGFs play a major role in the unregulated growth and metastasis of RMS. In vitro, IGF‐II and IGF‐I increased migration of RD cells to 124 ± 9% (P < 0.01) and 131 ± 8% (P < 0.05) of control, respectively. IGF‐II‐induced migration was abolished by insulin‐like growth factor binding protein‐6 (IGFBP‐6) (P < 0.01), a relatively specific inhibitor of IGF‐II, and by plasminogen activator inhibitor type 1 (PAI‐1) (P < 0.05). Aprotinin, a plasmin inhibitor, and mannosamine, which inhibits the synthesis of glycosylphosphatidylinositol (GPI), thereby preventing anchorage of GPI‐linked proteins such as uPAR to the cell membrane, also decreased IGF‐II‐ (P < 0.05 for both) but not IGF‐I‐induced migration. [Arg54,Arg55]IGF‐II and [Leu27]IGF‐II, which preferentially bind to the IGF‐I and IGF‐II/mannose‐6‐phosphate receptors (IGF‐II/M6PR), respectively, both induced RD cell migration to 146 ± 8% (P < 0.01) and 120 ± 7% (P < 0.05) of control, respectively. An anti‐uPAR anti‐serum reduced IGF‐II‐ and IGF‐I‐induced migration (P < 0.05 for both). An anti‐low density lipoprotein‐related protein (LRP) anti‐serum reduced IGF‐I‐induced migration (P < 0.05). IGF‐I and ‐II both increased specific 125I‐single chain uPA (scuPA) binding to RD cells in a dose‐dependent manner (P < 0.01). These results suggest involvement of the PA/plasmin system in IGF‐induced migration and indicate important roles these systems may have in RMS metastasis. J. Cell. Physiol. 197: 131–138, 2003© 2003 Wiley‐Liss, Inc.
Glutathione mediates LPS‐stimulated COX‐2 expression via early transient p42/44 MAPK activation
Authors: Chen Jian‐Xiong , Berry Leonard C. , Christman Brian W. , Meyrick Barbara
Published: August 2003

This study examines whether endotoxin (LPS)‐stimulated COX‐2 is modulated by an interaction between mitogen activated protein kinases (MAPK) and intracellular glutathione. Bovine pulmonary artery endothelial cells (BPAEC) were pretreated for 30 min with the following prior to addition of 0.1 μg/ml endotoxin in 2% FBS in medium 199: 5 mM N‐acetylcysteine (NAC) or 5 mM glutathione ethyl ester (GSE) (modulators of intracellular glutathione); 10 μM SB203580 or 25 μM PD98059 (inhibitors of p38 and p42/44 MAPKs, respectively). End‐points included assessment of COX‐1 and COX‐2 gene expression by reverse transcription polymerase chain reaction (RT‐PCR); COX‐1, COX‐2, p38, and p42/44 protein by Western analysis; and measurement of PGE2 and 6‐keto‐PGF1α releases by GC/MS. Both GSE and NAC resulted in significant exacerbation of the LPS‐stimulated increase in COX‐2 gene and protein expression and prostaglandin release, and suppressed the LPS‐induced decrease in COX‐1. LPS caused a biphasic activation of p42/44 MAPKs, an early increase peaking at 30 min and a second sustained phase, lasting up to 24 h; LPS also caused an early and sustained increase p38 MAPK activity. Pretreatment of cells with either GSE or NAC increased the early LPS‐stimulated activation of p42/44 but had little effect on the sustained phase. Inhibition of either p38 or p42/44 MAPKs suppressed LPS‐stimulated COX‐2 gene and protein expression, and prostaglandin release (P < 0.05) but had little effect on COX‐1. We conclude that intracellular glutathione modulates LPS‐stimulated COX‐2 gene expression and prostaglandin synthesis in BPAEC via early activation of p42/44 MAPKs. J. Cell. Physiol. 197: 86–93, 2003© 2003 Wiley‐Liss, Inc.
Sorsby fundus dystrophy mutation Timp3S156C affects the morphological and biochemical phenotype but not metalloproteinase homeostasis
Authors: Soboleva Galina , Geis Birgit , Schrewe Heinrich , Weber Bernhard H.F.
Published: August 2003

The tissue inhibitor of metalloproteinases‐3 (TIMP3) is a multifunctional protein tightly associated with the extracellular matrix (ECM). A specific type of mutation in TIMP3 which results in potentially unpaired cysteine residues at the C‐terminus of the protein has been shown to cause Sorsby fundus dystrophy (SFD), an autosomal dominant retinopathy of late onset. An early finding in SFD is a striking accumulation of protein and lipid material in Bruch's membrane, a multilayered ECM structure located between the choroid and the RPE. To study the molecular mechanisms underlying SFD pathology, we recently generated two mouse lines, one deficient in Timp3 (Timp3−/−) and one carrying an SFD‐related mutation in the orthologous murine Timp3 gene (Timp3S156C/S156C). We now established immortalized fibroblast cells from the mutant mouse strains and provide evidence that the various cell lines display distinct morphological and physiological features that are dependent on the mutational status of the Timp3 protein in the secreted ECM. We show that matrix metalloproteinase (MMP) activity and inhibitory properties of Timp3 are not affected by the SFD‐associated mutation. We further demonstrate that Timp3S156C protein accumulates in the ECM of the mutant fibroblast cells and that this accumulation is not due to a prolonged turnover rate of mutant vs. normal Timp3. We also show that the relative abundance of mutant and normal Timp3 in the ECM has no measurable effects on cellular phenotypes. Together, these findings suggest (i) a functional role of normal Timp3 in pathways determining cellular morphology and (ii) a loss of this particular function as a consequence of the Ser156Cys mutation. We therefore hypothesize that SFD pathogenesis is due to a loss‐of‐function mutation in TIMP3. J. Cell. Physiol. 197: 149–156, 2003© 2003 Wiley‐Liss, Inc.
Catalytic activity of Cdk9 is required for nuclear co‐localization of the Cdk9/cyclin T1 (P‐TEFb) complex
Authors: Napolitano Giuliana , Majello Barbara , Lania Luigi
Published: August 2003

Cdk9 and its binding partner cyclin T1 comprise the positive elongation factor b (P‐TEFb). P‐TEFb phosphorylates the RNA polymerase II carboxyl‐terminal‐domain (CTD) allowing efficient transcription elongation. Recent studies showed that Cdk9 is a predominant nuclear protein, and here we investigated the functional requirement for nuclear localization of Cdk9. We found that the catalytic inactive kinase mutant (Cdk9dn) fails to accumulate in the nucleus showing a diffuse sub‐cellular localization. In addition to the catalityc activity, nuclear localization of Cdk9 protein requires the presence of the phospho‐acceptor sites at the C‐terminus tail. Finally, enforced expression of wild‐type cyclinT1, which enhances nuclear localization of Cdk9wt, fails to direct the Cdk9 mutants to the nucleus. Collectively, these findings implicate that nuclear localization of Cdk9 requires auto‐phosphorylation of the kinase, and highlight the presence of a regulatory mechanism underlying the nuclear localization of the P‐TEFb complex. J. Cell. Physiol. 197: 1–7, 2003© 2003 Wiley‐Liss, Inc.
Process integration technology review: background and applications in the chemical process industry
Authors: Dunn Russell F , El‐Halwagi Mahmoud M
Published: August 2003

Process integration is a holistic approach to process design and operation which emphasizes the unity of the process. Process integration design tools have been developed over the past two decades to achieve process improvement, productivity enhancement, conservation in mass and energy resources, and reductions in the operating and capital costs of chemical processes. The primary applications of these integrated tools have focused on resource conservation, pollution prevention and energy management. Specifically, the past two decades have seen the development and/or application of process integration design tools for heat exchange networks (HENs), wastewater reduction and water conservation networks, mass exchange networks (MENs), heat‐ and energy‐induced separation networks (HISENs and EISENs), waste interception networks (WINs) and heat‐ and energy‐induced waste minimization networks (HIWAMINs and EIWAMINs), to name a few. This paper provides an overview of some of these developments and outlines major driving forces and hurdles. The fundamental aspects of this approach along with their incorporation in an overall design methodology will be discussed. The paper also highlights several recent applications of process integration to industrial processes. Copyright © 2003 Society of Chemical Industry
Kinetics of the degradation by catalytic hydrogenation of tyrosol, a model molecule present in olive oil waste waters
Authors: Richard Dominique , Delgado‐Nuñez María de Lourdes
Published: August 2003

Olive mill wastewaters (OMW) constitute an important environmental problem due, among other things, to their high phenolic content. The phytotoxicity of polyphenols makes them resistant to biological treatment. A three‐step process comprising adsorption–concentration, catalytic hydrogenation and regeneration on a fixed bed of adsorbent–catalyst was investigated to remove such compounds. Tyrosol (2‐(4‐hydroxyphenyl)‐ethanol) was taken as representative of the polyphenols present in OMW. The catalytic hydrogenation of tyrosol by 30 g kg−1 ruthenium/activated carbon catalyst was carried out in order to establish the reaction mechanism and kinetics necessary for the design of a pilot‐scale reactor. Total conversion of tyrosol into non‐aromatic compounds was achieved under mild conditions of temperature and pressure. The overall rate of tyrosol removal at 4 MPa and 353 K was $\hbox{11.26 g h}^{-1} \hbox{g}^{-1}_{\rm{catalyst}}$. Copyright © 2003 Society of Chemical Industry
Use of response surface methodology to evaluate the optimum environmental conditions for the bioconversion of xylose into xylitol in eucalyptus hemicellulosic hydrolysate
Authors: Almeida e Silva João B , Canilha Larissa , Canettieri Eliana V , Felipe Maria GA , Nápoles Solenzal Ana Irene
Published: August 2003

Candida guilliermondii fermented a eucalyptus hemicellulosic hydrolysate that had been treated with calcium oxide, phosphoric acid and activated charcoal. The influences of different xylose and ammonium sulfate concentrations on the production of xylitol were studied. A response surface was obtained with value R2 > 0.97 and p < 0.01. The model estimated a maximum xylitol production of 20.82 g dm−3 in the culture medium supplemented with ammonium sulfate (1.8 g dm−3) and xylose (45.5 g dm−3). Copyright © 2003 Society of Chemical Industry
Effect of surfactants on phenol removal by the method of polymerization and precipitation catalysed by Coprinus cinereus peroxidase
Authors: Sakurai Akihiko , Masuda Mina , Sakakibara Mikio
Published: August 2003

The removal of phenol by peroxidase‐catalysed polymerization was examined using Coprinus cinereus peroxidase in the presence of surfactants. The non‐ionic surfactants with poly(oxyethene) residues, Triton X‐100, Triton X‐405 and Tween 20, enhanced the phenol removal efficiency at a level similar to high relative molecular mass poly(ethylene glycol) (relative molecular mass 3000). Although the improvement in the removal efficiency was less than that of Triton X‐100, Span 20, sodium lauryl sulfate (SDS) and lauryl trimethylammonium bromide (DTAB) also enhanced the removal efficiency. The requirement of the enzyme for almost 100% removal of 100 mg dm−3 phenol decreased to one‐fourth by the addition of 30 mg dm−3 Triton X‐100. Triton X‐100, Triton X‐405, Tween 20 and DTAB could reactivate the enzyme precipitated with the phenol polymer, leading to the restarting of the phenol removal reaction. Copyright © 2003 Society of Chemical Industry
Removal of suspended solids and residual oil from palm oil mill effluent
Authors: Ahmad Abdul Latif , Ismail Suzylawati , Ibrahim Norliza , Bhatia Subhash
Published: August 2003

Palm oil mill effluent (POME) was pretreated to remove suspended solids and residual oil. The processes used were flocculation, solvent extraction, adsorption and membrane separation. Flocculation was used to remove suspended solids, and solvent extraction and adsorption processes were used to remove residual oil. Membrane separation was subsequently applied to remove any residual suspended solids and oil remaining after the pretreatments. The solvent extraction and adsorption processes were operated on a batch basis whereas membrane separation was performed in continuous mode. The treatment efficiency of the processes was measured as percentage removal of suspended solids and oil respectively. The optimum values of the process parameters obtained in the flocculation process were an alum dosage of 4000 mg dm−3, mixing speed of 150 rpm for 1 h and sedimentation time of 270 min, resulting in 93% suspended solids removal. In the solvent extraction process, a 95% reduction in residual oil was obtained using n‐hexane as a solvent with 20 min of mixing at 200 rpm. The ratio of solvent to POME was 6:10 and carried out at pH 9. In the batch adsorption process, an 88% reduction in residual oil was obtained at a mixing speed of 100 rpm for 1 h, pH 9 and an adsorbent dosage of 300 g dm−3. In membrane separation processes, GH and CE(GH) membranes gave 63% and 49% reductions in suspended solids and residual oil respectively at pH 9 and pressure of 1000 kPa. Copyright © 2003 Society of Chemical Industry
Mathematical model for growth process of a recombinant yeast having saccharification and fermentation activities
Authors: Kobayashi Fumihisa , Nakamura Yoshitoshi
Published: August 2003

A mathematical model for direct alcohol fermentation from starch was proposed using an amylase‐producing recombinant yeast, Saccharomyces cerevisiae SR93. This model consisted of the reaction rate equations for glucoamylase synthesis in the recombinant yeast, starch degradation by a glucoamylase, cell growth, production of glucose, and production of ethanol. The rate of glucoamylase synthesis was expressed on the basis of the diauxic growth model that represents catabolite repression and enzyme induction. The rate of starch degradation was expressed on the basis of the enzymatic hydrolysis model representing the change of structure resulting from starch degradation. The calculated values were in satisfactory agreement with the experimental data in a batch culture of direct alcohol fermentation from starch using S cerevisiae SR93. Furthermore, the calculated values obtained by changing only one parameter concerning the synthesis rate of glucoamylase were in satisfactory agreement with the experimental data using another recombinant yeast, S cerevisiae SR96. Copyright © 2003 Society of Chemical Industry
Drastic improvement of bioethanol recovery using a pervaporation separation technique employing a silicone rubber‐coated silicalite membrane
Authors: Ikegami Toru , Kitamoto Dai , Negishi Hideyuki , Haraya Kenji , Matsuda Hiroyuki , Nitanai You , Koura Nobuyuki , Sano Tsuneji , Yanagishita Hiroshi
Published: August 2003

A coupled fermentation/pervaporation process for reliable production of concentrated ethanol was studied using ethanol permselective silicalite membranes coated with two types of silicone rubber, KE‐45 and KE‐108, as a hydrophobic material. Ethanol recovery was greatly improved by using a membrane coated with KE‐45 silicone rubber. The recovered ethanol concentration in the permeate was 67% (w/w), and the amount of recovered ethanol from the broth was more than 10 times higher than that using a non‐coated membrane. Succinic acid and glycerol, by‐products created during fermentation, interfered with the pervaporation performance of the coated membrane when used to separate an ethanol/water solution. Copyright © 2003 Society of Chemical Industry
Resolving morphology and antibody labeling over large distances in tissue sections
Authors: Jacobs Marc D. , Donaldson Paul J. , Cannell Mark B. , Soeller Christian
Published: August 2003

Protein expression patterns are a primary determinant of tissue function and in this study we developed methods to study protein expression over macroscopic distances at subcellular levels of detail. Using the mammalian lens as a model tissue system, we show that by combining two‐photon microscopy with novel image montage methods (fast beam blanking coupled with mathematical alignment tools) we have extended the limited field of view of laser scanning microscopes. To illustrate the utility of our approach, the distribution of connexin‐46 was visualized across equatorial sections of the rat mammalian lens. By optimizing fixation protocols, good morphological preservation could be achieved over the thickness of the lens (∼4 mm) while preserving antigenicity of lens proteins. Using the same image data, changes in lens fiber cell morphology were mapped quantitatively by automatic image analysis routines. The methods presented should be generally applicable to any tissue system where changes in antibody labeling and tissue structure occur over large and small distances. Microsc. Res. Tech. 62:83–91, 2003. © 2003 Wiley‐Liss, Inc.
Development, principles, and applications of automated ice fabric analyzers
Authors: Wilen L.A. , Diprinzio C.L. , Alley R.B. , Azuma N.
Published: August 2003

We review the recent development of automated techniques to determine the fabric and texture of polycrystalline ice. The motivation for the study of ice fabric is first outlined. After a brief introduction to the relevant optical concepts, the classic manual technique for fabric measurement is described, along with early attempts at partial automation. Then, the general principles behind fully automated techniques are discussed. We describe in some detail the similarities and differences of the three modern instruments recently developed for ice fabric studies. Next, we discuss briefly X‐ray, radar, and acoustic techniques for ice fabric characterization. We also discuss the principles behind automated optical techniques to measure fabric in quartz rock samples. Finally, examples of new applications that have been facilitated by the development of the ice fabric instruments are presented. Microsc. Res. Tech. 62:2–18, 2003. © 2003 Wiley‐Liss, Inc.
SEM/EDS comparison of polar and seasonal temperate ice
Authors: Obbard Rachel , Iliescu Daniel , Cullen Daniel , Chang Janelle , Baker Ian
Published: August 2003

We have developed a method employing a scanning electron microscope (SEM) and energy dispersive X‐ray spectroscopy for examining uncoated ice specimens. By permitting the ice to sublimate in the SEM at temperatures between −115° and −60°C, enough ions are produced to prevent specimen charging. The absence of a conductive coating permits both the acquisition of uncompromised X‐ray spectra, and a dynamic view of impurities as the ice sublimates. The method has enabled us to examine the microstructure and impurities in ice in ways not possible through standard melt chemistry measurements or even through using a SEM to study coated samples. Soluble impurities appeared either as white spots in grain interiors, grain boundaries, and triple junctions, or as filaments in grain boundaries. Inclusions of insoluble impurities have also been observed in natural ice. Thus, we have been able to compare the microstructural location and concentration of impurities in ice from different terrestrial locations. Even when ion chromatography of the melt from two core sections reveals similar levels of impurities, the morphology and location of the impurity aggregates can be quite different. Analysis of impurity type and location can provide clues to the depositional environment and history of the ice. Microsc. Res. Tech. 62:49–61, 2003. © 2003 Wiley‐Liss, Inc.
Imaging dislocations in ice
Authors: Baker I.
Published: August 2003

Three techniques have been used to study dislocations in ice: etch pitting‐replication, transmission electron microscopy, and X‐ray topography (XT). It is shown that, because ice is a weak absorber of X‐rays and can be produced with a low dislocation density, allowing relatively thick specimens to be studied, the most useful technique is XT. The observations that have been made with conventional XT are briefly outlined. However, the introduction of high‐intensity synchrotron radiation, with its concomitant short exposure times, showed that images obtained through conventional XT observations were of dislocations that had undergone recovery. The important dynamic observations and measurements that have been made using synchrotron X‐ray topography are presented. Dynamic synchrotron X‐ray topography observations of ice single crystals undergoing deformation in situ have shown that slip mainly occurs by the movement of screw and 60° ⅓〈1120〉 dislocations on the basal plane, although non‐basal slip by edge dislocations can also occur. The operation of Frank‐Read and other dislocation multiplication sources have been clearly demonstrated and dislocation velocities have been measured. In contrast, in polycrystals, dislocation generation occurred at grain boundaries where there are stress concentrations before lattice dislocation generation mechanisms operate. Faulted dislocation loops have been determined to be mainly interstitial in both polycrystals and single crystals. Microsc. Res. Tech. 62:70–82, 2003. © 2003 Wiley‐Liss, Inc.
Collecting, shipping, storing, and imaging snow crystals and ice grains with low‐temperature scanning electron microscopy
Authors: Erbe Eric F. , Rango Albert , Foster James , Josberger Edward G. , Pooley Christopher , Wergin William P.
Published: August 2003

Methods to collect, transport, and store samples of snow and ice have been developed that enable detailed observations of these samples with a technique known as low‐temperature scanning electron microscopy (LTSEM). This technique increases the resolution and ease with which samples of snow and ice can be observed, studied, and photographed. Samples are easily collected in the field and have been shipped to the electron microscopy laboratory by common air carrier from distances as far as 5,000 miles. Delicate specimens of snow crystals and ice grains survive the shipment procedures and have been stored for as long as 3 years without undergoing any structural changes. The samples are not subjected to the melting or sublimation artifacts. LTSEM allows individual crystals to be observed for several hours with no detectable changes. Furthermore, the instrument permits recording of photographs containing the parallax information necessary for three‐dimensional imaging of the true shapes of snowflakes, snow crystals, snow clusters, ice grains, and interspersed air spaces. This study presents detailed descriptions of the procedures that have been used successfully in the field and the laboratory to collect, ship, store, and image snow crystals and ice grains. Microsc. Res. Tech. 62:19–32, 2003. Published 2003 Wiley‐Liss, Inc.
Snow metamorphism as revealed by scanning electron microscopy
Authors: Dominé Florent , Lauzier Thomas , Cabanes Axel , Legagneux Loïc , Kuhs Werner F. , Techmer Kirsten , Heinrichs Till
Published: August 2003

Current theories of snow metamorphism indicate that sublimating snow crystals have rounded shapes, while growing crystals have shapes that depend on growth rates. At slow growth rates, crystals are rounded. At moderate rates, they have flat faces with rounded edges. At fast growth rates, crystals have flat faces with sharp edges, and they have hollow faces at very fast growth rates. The main growth/sublimation mechanism is thought to be by the homogeneous nucleation of new layers at or near crystal edges. It was also suggested that the equilibrium shape of snow crystals would be temperature dependent: rounded above −10.5°C, and faceted below. To test these paradigms, we have performed SEM investigations of snow samples having undergone metamorphism under natural conditions, and of snow samples subjected to isothermal metamorphism at −4° and −15°C in the laboratory. In general, current theories predicting crystal shapes as a function of growth rates, and of whether crystals are growing or sublimating, are verified. However, the transition in equilibrium shapes from rounded to faceted at −10.5°C is not observed in our isothermal experiments that reveal a predominance of rounded shapes after more than a month of metamorphism at −4 and −15°C. Some small crystals with flat faces that also have sharp angles at −15°C, are observed in our isothermal experiments. These faces are newly formed, and contradict current theory. Several hypotheses are proposed to explain their occurrence. One is that they are due to sublimation at emerging dislocations. Microsc. Res. Tech. 62:33–48, 2003. © 2003 Wiley‐Liss, Inc.
SEM studies of the morphology and chemistry of polar ice
Authors: Barnes Piers R.F. , Wolff Eric W. , Mallard David C. , Mader Heidy M.
Published: August 2003

Determining the microphysical location of impurities in natural ice from the polar regions is necessary for understanding the physical properties of ice and for assuring the integrity of ice core records. SEM, using a cold stage and X‐ray microanalytical techniques, has proved to be the most powerful method so far for undertaking such work. Methods are adapted from those used to study frozen hydrated biological material. Sublimation within the cryo‐chamber is often needed in order to concentrate impurities onto a plane, but this can lead to artifacts that must be recognized. Over 100 samples from different depths and sites in Greenland and Antarctica have been examined. Typical physical features, including air bubbles, clathrate hydrates of air, and dust particles are identified. The dust is found preferentially at grain boundaries in some samples; by pinning the boundaries, it can slow grain growth. Of the soluble material, chloride seems to be found most frequently in the ice lattice. Other impurities are found at grain boundaries, and only when the bulk concentration exceeds a threshold, at triple junctions. These findings give new insights into processes determining the physical properties of ice samples and of ice sheets, and new impetus for theoretical studies of the energetics that lead to this distribution. Microsc. Res. Tech. 62:62–69, 2003. © 2003 Wiley‐Liss, Inc.
Synthesis of 13C3‐hydroxyacetone
Authors: Dicus Christopher W. , Burnham Kevin J. , Judith Charles M. , Nantz Michael H.
Published: August 2003

13C3‐Hydroxyacetone is prepared in three steps from 13C2‐2‐bromoacetic acid. Bromide displacement by sodium p‐methoxybenzyl alkoxide followed by treatment of the carboxylic acid with 13C‐methyl lithium furnishes PMB‐protected hydroxyacetone. Deprotection using DDQ delivers the title compound. Copyright © 2003 John Wiley & Sons, Ltd.
Synthesis of tracer labelled [11,12‐3H]‐β‐carotene
Authors: Goswami Bhabesh C. , Barua Arun B.
Published: August 2003

The synthesis of tracer labelled [11,12‐3H]‐β‐carotene is described. The procedure uses Wittig condensation of tracer labelled 3H‐retinal (retinal spiked with [11,12‐3H]‐retinal) with retinyl triphenylphosphonium bromide. The preparation of tracer labelled[3H]‐β‐carotene is suitable for studies involving bioavailability and bioconversion of β‐carotene to vitamin A. Copyright © 2003 John Wiley & Sons, Ltd.
A facile and efficient synthesis of 14C‐labelled sulforaphane
Authors: D'Souza Christopher A. , Amin Shantu , Desai Dhimant
Published: August 2003

Isothiocyanates have gained considerable attention for their role as potent chemopreventive agents. Sulforaphane, 1a (SFN), a naturally occurring isothiocyanate, was isotopically labelled in five steps starting from 3‐(methylthio)‐1‐propanol (2). Reacting 2 with tosyl chloride in the presence of Et3N yielded the tosylate 3. Gently refluxing 3 with K14CN in DMF gave the nitrile 4b. Reduction to the amine 5b was achieved using BH3‐THF. Oxidation with 30% hydrogen peroxide followed by treatment with thiophosgene yielded (±)[1‐14C]SFN, 1b. The overall radiochemical yield was 4.4% based on the starting K14CN. Copyright © 2003 John Wiley & Sons, Ltd.
Synthesis of tritium and carbon‐14 labelled NNC 61‐4655: a potent dual‐acting PPARα and PPARγ agonist
Authors: Johansen Steen K. , Martiny Lars
Published: August 2003

The synthesis of the potent dual‐acting PPARα and PPARγ agonist NNC 61‐4655 labelled with tritium and carbon‐14 is reported. Tritium labelled NNC 61‐4655 was obtained in three steps with introduction of tritium through catalytic tritium‐halogen exchange of an aryl bromide precursor. This provided [3H]NNC 61‐4655 in 39% overall radiochemical yield with a specific activity of 49 Ci/mmol. Carbon‐14 labelled NNC 61‐4655 was obtained in five steps starting from bromo[1‐14C]acetic acid. The synthetic sequence, which included a Horner–Wadsworth–Emmons olefination and a Mitsunobu alkylation, provided [14C]NNC 61‐4655 in 33% overall radiochemical yield with a specific activity of 57.4 mCi/mmol. Copyright © 2003 John Wiley & Sons, Ltd.
Synthesis of the serotonin transporter ligand (±)‐10‐methyl 3‐[6‐nitro‐(2‐quinolinyl)]‐3,10‐diazabicyclo‐[4.3.1]‐decane ([11C‐methyl]NS 2495) and first in vivo results
Authors: Audrain Hélène , Bender Dirk , Scheel‐Krüger Jørgen , Nielsen Elsebet Ø. , Olsen Gunnar M. , Peters Dan , Cumming Paul
Published: August 2003

The serotonin transporter ligand (±)‐10‐[11C]‐methyl 3‐[6‐nitro‐(2‐quinolinyl)]‐3,10‐diazabicyclo‐[4.3.1]‐decane ([11C‐methyl]NS 2495) was synthesized via a methylation reaction with [11C]methyl iodide. The radiochemical purity exceeded 99% and the specific radioactivity was found to be 1.8 GBq/μmol at 40 min after the end of bombardment. The uptake of the tracer in the brain of a living pig was recorded by positron emission tomography (PET), first in a baseline condition, and again after treatment with citalopram (1 mg/kg, i.v.) to displace the specific binding. The distribution volume relative to the metabolism‐corrected arterial input was high in pig brain, ranging from 75–150 ml g−1; treatment with citalopram uniformly reduced the distribution volume to 75 ml g−1. Binding potential (pB) maps generated using the cerebellum as a reference tissue showed highest binding in the mesencephalon and cingulate cortex, where the magnitude of pB was close to 0.6. Thus, the pattern of binding in vivo agrees with the known pattern of serotonin innervations in pig brain. However, the specific binding was incompletely displaced by pre‐treatment with citalopram. Thus, [11C‐methyl]NS 2495 can label serotonin transporters in a PET study of the brain of a living pig, but full displacement by cold citalopram was not obtained in vivo, possibly reflecting binding sites which are inaccessible to citalopram. Copyright © 2003 John Wiley & Sons, Ltd.