bluuebunny/crossref_metadata_2025 huggingface dataset:
Differentiation‐dependent induction of CYP1A1 in cultured rat small intestinal epithelial cells, colonocytes, and human colon carcinoma cells: Basement membrane‐mediated apoptosis
Authors: Sterling Kenneth M. , Cutroneo Kenneth R.
Published: October 2002

Rat small intestinal epithelial cells and human colon adenocarcinoma cells cultured on Matrigel™ expressed the differentiation specific enzyme, sucrase‐isomaltase, as determined by indirect immunofluorescence. Rat small intestinal epithelial cells, rat colonocytes, and human colon adenocarcinoma cells developed an altered morphology when cultured on Matrigel and became apoptotic within 24–48 h. Benzo[a]pyrene and 2,3,7,8‐tetrachlorodibenzo‐p‐dioxin caused a 2‐ and 5‐fold induction, respectively, of ethoxyresorufin‐o‐deethylase activity in rat small intestinal epithelial cells cultured on Matrigel. Benzo[a]pyrene‐ or 2,3,7,8‐tetrachlorodibenzo‐p‐dioxin‐induced ethoxyresorufin‐o‐deethylase activity in rat small intestinal epithelial cells cultured on plastic was not detected. 2,3,7,8‐tetrachlorodibenzo‐p‐dioxin treatment caused a 14‐fold induction of transfected, rat CYP1A1‐promoter‐luciferase activity in rat small intestinal epithelial cells cultured on Matrigel. Benzo[a]pyrene and 2,3,7,8‐tetrachlorodibenzo‐p‐dioxin treatment induced ethoxyresorufin‐o‐deethylase activity by 6‐ and 1.6‐fold, respectively in rat colonocytes cultured on Matrigel. Induction of ethoxyresorufin‐o‐deethylase activity was not observed in rat colonocytes cultured on plastic. CYP1A1‐promoter‐luciferase activity was induced 3‐fold by 2,3,7,8‐tetrachlorodibenzo‐p‐dioxin in rat colonocytes cultured on Matrigel. Induction of CYP1A1‐promoter‐luciferase activity in rat small intestinal epithelial cells or rat colonocytes cultured on plastic was not observed. Ethoxyresorufin‐o‐deethylase activity in human colon adenocarcinoma cells, cultured on either plastic or Matrigel, was induced 7‐fold by benzo[a]pyrene. 2,3,7,8‐Tetrachlorodibenzo‐p‐dioxin‐induced ethoxyresorufin‐o‐deethylase activity was 2‐fold greater in human colon adenocarcinoma cells cultured on Matrigel compared to cells cultured on plastic. Extracellular matrix‐mediated differentiation and apoptosis of intestinal cells provide in vitro systems for study of the regulation of CYP1A1 expression, carcinogen activation in the gut and mechanism(s) of apoptosis of colon cancer cells. J. Cell. Biochem. 86: 440–450, 2002. © 2002 Wiley‐Liss, Inc.
Synergistic stimulation of MUC1 expression in normal breast epithelia and breast cancer cells by interferon‐γ and tumor necrosis factor‐α
Authors: Lagow Errin L. , Carson Daniel D.
Published: October 2002

The MUC1 gene encodes a transmembrane mucin glycoprotein that is overexpressed in human breast cancers. Persistent stimulation by proinflammatory cytokines may contribute to increased MUC1 transcription by tumor cells. We demonstrate that MUC1 expression in T47D breast cancer cells and normal human mammary epithelial cells (HMEC) is enhanced by tumor necrosis factor‐α (TNF‐α) in the presence of interferon‐γ (IFN‐γ). MUC1 responsiveness to these cytokines was modest in T47D cells and robustly induced in HMEC. Transient transfection of T47D cells with mutant MUC1 promoter constructs revealed that a κB site at −589/−580 and the STAT‐binding element at −503/−495 and were required for cooperative stimulation by TNFα and IFN‐γ. Binding of NFκB p65 to the MUC1 κB site was induced by TNF‐α treatment, as demonstrated by electrophoretic mobility shift assay. Specific mutation of the κB site prevented binding of NFκB p65 and blocked TNF‐α stimulation of MUC1 promoter activity. Collectively, these studies demonstrate synergistic stimulation of MUC1 expression by TNF‐α and IFN‐γ that is mediated by independent actions of NFκB p65 and STAT1α upon κB and STAT sites, respectively, in the MUC1 promoter. Strong induction of MUC1 expression by these proinflammatory cytokines is clearly evident in normal mammary epithelium. In contrast, breast tumor cells appear to override normal regulatory responses via as yet undefined cis‐elements. J. Cell. Biochem. 86: 759–772, 2002. © 2002 Wiley‐Liss, Inc.
Biochemical and cellular characteristics of the four splice variants of protein kinase CK1α from zebrafish (Danio rerio)
Authors: Burzio Veronica , Antonelli Marcelo , Allende Catherine C. , Allende Jorge E.
Published: October 2002

Protein kinase CK1 (previously known as casein kinase I) conforms to a subgroup of the great protein kinase family found in eukaryotic organisms. The CK1 subgroup of vertebrates contains seven members known as α, β, γ1, γ2, γ3, δ, and ε. The CK1α gene can generate four variants (CK1α, CK1αS, CK1αL, and CK1αLS) through alternate splicing, characterized by the presence or absence of two additional coding sequences. Exon “L” encodes a 28‐amino acid stretch that is inserted after lysine 152, in the center of the catalytic domain. The “S” insert encodes 12 amino acid residues and is located close to the carboxyl terminus of the protein. This work reports some biochemical and cellular properties of the four CK1α variants found to be expressed in zebrafish (Danio rerio). The results obtained indicate that the presence of the “L” insert affects several biochemical properties of CK1α: (a) it increases the apparent Km for ATP twofold, from ∼30 to ∼60 μM; (b) it decreases the sensitivity to the CKI‐7 inhibitor, raising the I50 values from 113 to ∼230 μM; (c) it greatly decreases the heat stability of the enzyme at 40°C. In addition, the insertion of the “L” fragment exerts very important effects on some cellular properties of the enzyme. CK1αL concentrates in the cell nucleus, excluding nucleoli, while the CK1α variant is predominantly cytoplasmic, although some presence is observed in the nucleus. This finding supports the thesis that the basic‐rich region found in the “L” insert acts as a nuclear localization signal. The “L” insert‐containing variant was also found to be more rapidly degraded (half‐life of 100 min) than the CK1α variant (half‐life of 400 min) in transfected Cos‐7 cells. J. Cell. Biochem. 86: 805–814, 2002. © 2002 Wiley‐Liss, Inc.
Sp1 and Sp3 activate the testis‐specific histone H1t promoter through the H1t/GC‐box
Authors: Wilkerson Donald C. , Wolfe Steven A. , Grimes Sidney R.
Published: October 2002

The testis‐specific linker histone H1t gene is transcribed exclusively in mid to late pachytene primary spermatocytes. Tissue specific expression of the gene is mediated in large part through elements located within the proximal promoter. Previous work in transgenic animals showed that a unique 40 bp promoter element designated H1t/TE is essential for spermatocyte‐specific expression. The H1t/TE element contains a GC‐box, which is a perfect consensus binding site for members of the Sp family of transcription factors. We have shown that Sp1 and Sp3 are present in testis cells from 9‐day‐old and adult rats and in pachytene primary spermatocytes and early spermatids and that they can bind to the H1t/GC‐box. Mutagenesis of the GC‐box reduced H1t promoter activity. Furthermore, a CpG dinucleotide within the GC‐box was totally unmethylated in rat testis primary spermatocytes where the gene is transcribed but it was methylated in liver where the gene is silenced. These previous studies supported the importance of the GC‐box and suggested that Sp transcription factors contribute to expression of the H1t gene. In this study, we show that co‐transfection of Sp1 and Sp3 expression vectors leads to an upregulation of histone H1t promoter activity in several cell lines including testis GC‐2spd cells. However, very low H1t promoter activity is seen in GC‐2spd cells grown at 39°C, which correlates with lower levels of Sp1 and Sp3 in these cells grown at this elevated temperature. Upregulation of the H1t promoter by Sp1 and Sp3 was also seen in cotransfected NIH3T3 and C127I cell lines. On the other hand, co‐transfection of the Sp1 and Sp3 expression vectors does not lead to upregulation of activity of the cell‐cycle dependent histone H1d promoter. J. Cell. Biochem. 86: 716–725, 2002. © 2002 Wiley‐Liss, Inc.
MM3 parametrization of four‐ and five‐coordinated rhenium complexes by a genetic algorithm—Which factors influence the optimization performance?
Authors: Strassner Thomas , Busold Markus , Herrmann Wolfgang A.
Published: October 2002

Genetic algorithms (GA) were used to solve one of the multidimensional problems in computational chemistry, the optimization of force field parameters. The correlation between the composition of the GA, its parameters (pc,pm) and the quality of the results were investigated. The composition was studied for all combinations of a Simple GA/Steady State GA with a Roulette Wheel/Tournament Selector using different values each for crossover (0.5, 0.7, 0.9) and mutation rates (0.01, 0.02, 0.05, 0.10, 0.20). The results show that the performance is strongly dependent on the GA scheme, where the Simple GA/Tournament Selector yields the best results. Two new MM3 parameters were introduced for rhenium compounds with coordination number four (204) and coordination number five (205), the formal oxidation states of rhenium ranging from +V to +VII. A manifold of parameters (Re‐C, N, O, S) was obtained by using a diverse set of CSD structures. The advantage of the GA vs. UFF calculations is shown by comparison of several examples. The GA optimized parameters were able to reproduce the geometrical data of the X‐ray structures. © 2002 Wiley Periodicals, Inc. J Comput Chem 23: 282–290, 2002
Chemical shift driven geometry optimization
Authors: Witter Raiker , Prie Wolfram , Sternberg Ulrich
Published: October 2002

A new method for refinement of 3D molecular structures by geometry optimization is presented. Prerequisites are a force field and a very fast procedure for the calculation of chemical shifts in every step of optimization. To the energy, provided by the force field (COSMOS force field), a pseudoenergy, depending on the difference between experimental and calculated chemical shifts, is added. In addition to the energy gradients, pseudoforces are computed. This requires the derivatives of the chemical shifts with respect to the coordinates. The pseudoforces are analytically derived from the integral expressions of the bond polarization theory. Single chemical shift values attributed to corresponding atoms are considered for structural correction. As a first example, this method is applied for proton position refinement of the D‐mannitol X‐ray structure. A crystal structure refinement with 13C chemical shift pseudoforces is carried out. © 2002 Wiley Periodicals, Inc. J Comput Chem 23: 298–305, 2002
Efficient linear algebra routines for symmetric matrices stored in packed form
Authors: Ahlrichs Reinhart , Tsereteli Kakha
Published: October 2002

Quantum chemistry methods require various linear algebra routines for symmetric matrices, for example, diagonalization or Cholesky decomposition for positive matrices. We present a small set of these basic routines that are efficient and minimize memory requirements. © 2002 Wiley Periodicals, Inc. J Comput Chem 23: 306–309, 2002
Binding of D‐ and L‐captopril inhibitors to metallo‐β‐lactamase studied by polarizable molecular mechanics and quantum mechanics
Authors: Antony Jens , Gresh Nohad , Olsen Lars , Hemmingsen Lars , Schofield Christopher J. , Bauer Rogert
Published: October 2002

The bacterial Zn2+ metallo‐β‐lactamase from B. fragilis is a zinc‐enzyme with two potential metal ion binding sites. It cleaves the lactam ring of antibiotics, thus contributing to the acquired resistance of bacteria against antibiotics. The present study bears on the binuclear form of the enzyme. We compare several possible binding modes of captopril, a mercaptocarboxamide inhibitor of several zinc‐metalloenzymes. Two diastereoisomers of captopril were considered, with either a D‐ or an L‐proline residue. We have used the polarizable molecular mechanics procedure SIBFA (Sum of Interactions Between Fragments ab initio computed). Two β‐lactamase models were considered, encompassing 104 and 188 residues, respectively. The energy balances included the inter and intramolecular interaction energies as well as the contribution from solvation computed using a continuum reaction field procedure. The thiolate ion of the inhibitor is binding to both metal ions, expelling the bridging solvent molecule from the uncomplexed enzyme. Different competing binding modes of captopril were considered, either where the inhibitor binds in a monodentate mode to the zinc cations only with its thiolate ion, or in bidentate modes involving additional zinc binding by its carboxylate or ketone carbonyl groups. The additional coordination by the inhibitor's carboxylate or carbonyl group always occurs at the zinc ion, which is bound by a histidine, a cysteine, and an aspartate side chain. For both diastereomers, the energy balances favor monodentate binding of captopril via S−. The preference over bidentate binding is small. The interaction energies were recomputed in model sites restricted to captopril, the Zn2+ cations, and their coordinating end side chains from β‐lactamase (98 atoms). The interaction energies and their ranking among competing arrangements were consistent with those computed by ab initio HF and DFT procedures. © 2002 Wiley Periodicals, Inc. J Comput Chem 23: 1281–1296, 2002
Combined ab initio/empirical approach for optimization of Lennard‐Jones parameters for polar‐neutral compounds
Authors: Chen I Jen , Yin Daxu , MacKerell Alexander D.
Published: October 2002

The study of small functionalized organic molecules in aqueous solution is a useful step toward gaining a basic understanding of the behavior of biomolecular systems in their native aqueous environment. Interest in studying amines and fluorine‐substituted compounds has risen from their intrinsic physicochemical properties and their prevalence in biological and pharmaceutical compounds. In the present study, a previously developed approach which optimizes Lennard‐Jones (LJ) parameters via the use of rare gas atoms combined with the reproduction of experimental condensed phase properties was extended to polar–neutral compounds. Compounds studied included four amines (ammonia, methylamine, dimethylamine, and trimethylamine) and three fluoroethanes (1‐fluoroethane, 1,1‐difluoroethane, and 1,1,1‐trifluoroethane). The resulting force field yielded heats of vaporization and molecular volumes in excellent agreement with the experiment, with average differences less than 1%. The current amine CHARMM parameters successfully reproduced experimental aqueous solvation data where methylamine is more hydrophilic than ammonia, with hydrophobicity increasing with additional methylation on the nitrogen. For both the amines and fluoroethanes the parabolic relationship of the extent of methylation or fluorination, respectively, to the heats of vaporization were reproduced by the new parameters. The present results are also discussed with respect to the impact of parameterization approach to molecular details obtained from computer simulations and to the unique biological properties of fluorine in pharmaceutical compounds. © 2002 Wiley Periodicals, Inc. J Comput Chem 23: 199–213, 2002
Psychotherapists' representations of their patients
Authors: Geller Jesse D. , Lehman Adam K. , Farber Barry A.
Published: October 2002

Using a new measure, the Patient Representation Inventory (PRI), this study investigated the nature of psychotherapists' working clinical models of their patients. The data provided by 73 therapists suggest that, regardless of experience level or theoretical orientation (cognitive‐behavioral or psychodynamic), therapists tend to evoke such representations by accessing the words spoken during sessions and by recreating visual images of their patients' nonverbal contributions to the therapeutic dialogue. These representations tend to be in the service of more deeply understanding patients; their affective tone is suggestive of empathy, authenticity, and involvement. More‐experienced therapists tend to evoke representations that rely largely on bodily sensations and the emotional atmosphere reflective of clinical interactions. Apparently, similarities and differences among therapists can be studied not only at the manifest level of clinical work, but at the representational level as well. © 2002 Wiley Periodicals, Inc. J Clin Psychol 58: 733‐745, 2002.
Personality and deviant sexual fantasies: An examination of the MMPIs of sex offenders
Authors: Curnoe Suzanne , Langevin Ron
Published: October 2002

A sample of 228 sex offenders and nonsex offender controls were divided into two groups, based on the presence or absence of deviant sexual fantasies, as determined by their replies to the Clarke Sex History Questionnaire Fantasy Scales. The sex offenders were divided into six groups based on sexual preferences and criminal history. There were 14 heterosexual pedophiles, 23 homosexual pedophiles, 51 exhibitionists, 17 incest offenders, 24 sexual aggressives, and 57 multiple/miscellaneous offenders as well as 42 nonviolent nonsex offender controls. A 2 (fantasy group) × 7 (sex offender group) multivariate analysis with age and education as covariates was used to compare the MMPI validity scales (L, F, and K) and the 10 clinical scales as dependent variables. Deviant fantasizers had more clinically significantly scores on the F, Psychopathic Deviate, Masculinity‐Femininity, Paranoia, and Schizophrenia Scales compared to nondeviant fantasizers, regardless of group membership. Results suggest that deviant fantasizers may be more socially alienated and less emotionally stable than nondeviant fantasizers, although few were diagnosed as psychotic. The sex offender and control groups showed considerable overlap on mean MMPI Scales. The groups tended to differ in level of depression and extent of persecutory ideas, but the multivariate sex offender group effect was not statistically significant when problems of inhomogeneity of variance were corrected. There was no interaction of fantasy groups and sex offender groups, indicating that the two factors can be considered independently. © 2002 Wiley Periodicals, Inc. J Clin Psychol 58: 803‐815, 2002.
p115 Rho GTPase activating protein interacts with MEKK1
Authors: Christerson Lori B. , Gallagher Ewen , Vanderbilt Colleen A. , Whitehurst Angelique W. , Wells Clark , Kazempour Roxana , Sternweis Paul C. , Cobb Melanie H.
Published: October 2002

Mammalian MAP/ERK kinase kinase 1 (MEKK1) was identified as a mammalian homolog of Ste11p of the yeast pheromone‐induced mating pathway. Like Ste11p, MEKK1 is a MAP3 kinase linked to at least two MAP kinase cascades and regulatory events that require cytoskeletal reorganization. MEKK1 is activated by molecules that impact cytoskeletal function. MEKK1−/−cells are defective in cell migration, demonstrating that it is required for cell motility. MEKK1 has a 1,200 residue N‐terminal regulatory domain that interacts with a dozen identified proteins. Using part of the MEKK1 N‐terminus in a yeast two‐hybrid screen, we discovered a novel interaction with p115 Rho GTPase‐activating protein (GAP). The p115 Rho GAP binds to MEKK1 in vitro and in intact cells. The p115 Rho GAP has selectivity for RhoA over other Rho family members. Expression of p115 Rho GAP reduces MEKK1‐induced signaling to AP‐1. The reduced activation of AP‐1 is dependent on the association of MEKK1 with p115 Rho GAP, because deletion of the Rho GAP SH3 domain, which abrogates their interaction, restores the stimulatory effect of MEKK1 on AP‐1 activity. Here we have identified an MEKK1 binding partner that offers a connection between this protein kinase and the machinery regulating cytoskeletal reorganization. © 2002 Wiley‐Liss, Inc.
CDK9 has the intrinsic property to shuttle between nucleus and cytoplasm, and enhanced expression of CyclinT1 promotes its nuclear localization
Authors: Napolitano Giuliana , Licciardo Paolo , Carbone Roberta , Majello Barbara , Lania Luigi
Published: October 2002

CDK9 in association with cyclin T constitutes the P‐TEFb complex that stimulates transcription elongation of RNAPII transcripts by phosphorylation of the CTD of RNAPII. Here we report subcellular distribution of P‐TEFb in terms of localization of CDK9 and cyclin T1. We found that cyclin T1 is exclusively nuclear and it is present in nuclear‐speckled structures. CDK9, albeit mainly nuclear, was also visualized in the cytoplasm. We determined that CDK9 is actively exported from the nucleus, and that leptomycin B (LMB), a specific inhibitor of nuclear export, inhibits this process. Interestingly, enforced expression of cyclin T1 enhances nuclear localization of CDK9. These findings reveal a novel control mechanism for the function of the P‐TEFb complex. © 2002 Wiley‐Liss, Inc.
Prognostic significance of p16INK4aalterations and 9p21 loss of heterozigosity in locally advanced laryngeal squamous cell carcinoma
Authors: Bazan Viviana , Zanna Ines , Migliavacca Manuela , Sanz‐Casla Maria Teresa , Maestro Maria Luisa , Corsale Simona , Macaluso Marcella , Dardanoni Gabriella , Restivo Salvatore , Quintela Paloma López , Bernaldez Ricardo , Salerno Sergio , Morello Vincenza , Tomasino Rosa Maria , Gebbia Nicola , Russo Antonio
Published: October 2002

Thep16INK4agene, localized within chromosome 9p21, has been identified as a cyclin‐dependent kinase inhibitor and may negatively regulate the cell cycle acting as a tumor suppressor. Genetic alterations involving the 9p21 region are common in human cancers. A consecutive series of 64 untreated patients (median of follow up 53 months) undergoing surgical resection for locally advanced laryngeal squamous‐cell carcinomas (LSCCs) has been studied prospectively. Our purpose was to investigate p16 alterations (9p21 allelic loss, hypermethylation and point mutations) and their possible association with clinico‐pathological data and flow cytometric variables (DNA‐ploidy and S‐phase fraction (SPF)), and to determine the possible prognostic role of this gene in these tumors. PCR‐based techniques were used for investigating 9p21 loss of heterozygosity (LOH) and methylation promoter status of thep16gene. p16 mutations were detected by PCR‐SSCP (single strand conformation polymorphism) and sequencing. 9p21 LOH was detected in 16/62 (26%) informative tumors, point mutations in 5% (3/64) and hypermethylation in 9% (6/64) of the cases. p16 alterations were significantly associated with high SPF and DNA‐aneuploidy. By univariate analysis, poor histologic differentiation, stage IV, DNA‐aneuploidy and p16 point mutations proved to be significantly related to quicker relapse, whereas these same factors, and in addition high SPF, 9p21 LOH and any p16 alterations were significantly related to shorter overall survival. By Cox proportional hazards analysis only histologic grade (G3) and p16 point mutations were independently related to both disease relapse and death. Our study has identified p16 point mutations as important biomolecular indicators in LSCCs. © 2002 Wiley‐Liss, Inc.
Exendin‐4 differentiation of a human pancreatic duct cell line into endocrine cells: Involvement of PDX‐1 and HNF3β transcription factors
Authors: Zhou Jie , Pineyro Marco A. , Wang Xiaolin , Doyle Maire E. , Egan Josephine M.
Published: October 2002

Exendin‐4 (EX‐4), a long acting agonist of GLP‐1, induces an endocrine phenotype in Capan‐1 cells. Under culture conditions which include serum, ∼10% of the cells contain insulin and glucagon. When exposed to EX‐4 (0.1 nM, up to 5 days), the number of cells containing insulin and glucagon increased to ∼40%. Western blot analysis detected a progressive increase in protein levels of glucokinase and GLUT2 over 3 days of EX‐4 treatment. We explored the sequence of activation of certain transcription factors known to be essential for the beta cell phenotype: PDX‐1, Beta2/NeuroD, and hepatocyte nuclear factor 3β (HNF3β). Double immunostaining showed that PDX‐1 coexisted with insulin and glucagon in EX‐4‐treated cells. Treatment caused an increase in PDX‐1 protein levels by 24 h and induced its nuclear translocation. Beta2/NeuroD protein levels also increased progressively over 24 h. HNF3β protein level increased twofold as early as 6 h after EX‐4 treatment. EMSA results indicated that EX‐4 caused a 12‐fold increase in HNF3β binding to PDX‐1 promoter area II. Beta2/NeuroD protein levels progressively increased after 24 h treatment. Differentiation to insulin‐producing cells was also seen when Capan‐1 cells were transfected with pdx‐1, with 80% of these cells expressing insulin 3 days after transfection. PDX‐1 antisense totally inhibited such conversion. During the differentiation of duct cells to endocrine cells, cAMP levels (EX‐4 is a ligand for the GLP‐1, G‐protein coupled receptor) and MAP kinase activity increased. Our results indicate that EX‐4 activates adenylyl cyclase and MAP kinase which, in turn, may lead to activation of transcription factors necessary for an endocrine phenotype. Published 2002 Wiley‐Liss, Inc.
Neuronal regulation of bone metabolism and anabolism: Calcitonin gene‐related peptide‐, substance P‐, and tyrosine hydroxylase‐containing nerves and the bone
Authors: Imai Shinji , Matsusue Yoshitaka
Published: October 2002

Bone alters its metabolic and anabolic activities in response to the variety of systemic and local factors such as hormones and growth factors. Classical observations describing abundance of the nerves fibers in bone also predict a paradigm that the nervous system influences bone metabolism and anabolism. Identification of the nerve‐derived signaling molecules, capable of modulating cellular activities of the bone cells, facilitates a novel approach to study the biology of skeletal innervation. Many of the signaling molecules that may act as efferent agents on the bone cells fall into the category of neuropeptides. The present article reviews current understanding of the skeletal innervation and their proposed physiological effects on bone metabolism, with a special interest to calcitonin gene‐related peptide (CGRP)‐containing nerves fibers. CGRP is abundantly distributed in bone via sensory nerves, especially in the epiphyseal trabecular bones. Its in vitro actions to the cultured osteoblasts and osteoclasts, together with its in vivo localization, strongly support the paradigm that the nervous system influences bone metabolism. In addition, CGRP is recently shown to be expressed endogenously by the osteoblasts. Transgenic mice with osteoblasts overexpressing CGRP are characterized by increased bone formation rate and enhanced bone volume, suggesting that CGRP indeed acts on bone metabolism not only via nervous route but also via autocrine loop. The current article also reviews the distribution of nerve fibers containing substance P (SP), another sensory nerve‐specific neuropeptide, and tyrosine hydroxylase (TH), the rate‐limiting enzyme of catecholamine. The distinct effects of SP and catecholamines on the bone cells together with their in vivo influences manifested by experimental denervation studies suggest that the sensory and sympathetic nerves play important roles in bone metabolism. Microsc. Res. Tech. 58:61–69, 2002. © 2002 Wiley‐Liss, Inc.
Effect of hypothyroidism induced by propylthiouracil on ovarian function and structure in offspring from treated mothers (Rats)
Authors: Zertashia Akram , Jalali Samina , Ahmad Laiq , Mirza Asma
Published: October 2002

The aim of the present study was to investigate the effects of hypothyroidism induced during the pre‐ and postnatal periods of life on ovarian function and structure in offspring (pups) 120 days of age. Three groups were used. In the prenatal group, treatment was given from conception to parturition. In the postnatal group, treatment was given from parturition to 25 days postpartum. Hypothyroidism was induced by administration of 0.1% 6‐n‐propyl‐2‐thiouracil (PTU) in the drinking water of mothers. Body weights of the offspring were measured weekly. In each group, ten offspring were sacrificed at 120 days of age. Postnatal PTU treated pups showed delay in eye opening, teething, fur development, and weaning (35–37 days) compared to control animals (28–30 days). Body weight of offspring in the postnatal PTU treatment group was significantly decreased (P < 0.001), while the prenatal PTU treatment group showed a significant increase (P < 0.0001) compared to control animals. There was a significant (P < 0.05) reduction in paired ovarian weight of offspring in the postnatal PTU treatment group compared to control animals. Diameter of the ovaries was not affected by any treatment. Regarding the morphometery, only offspring in the prenatal PTU treatment group showed a significant (P < 0.001) increase in the diameter of graafian follicles. No significant difference was observed in morphometery of the granulosa layer, primary, and developing follicles of control and all treated groups. Number of primary, developing, and graafian follicles of all the treated groups was similar to that of the control group. The corpora lutea of the postnatal PTU treated group contained a population of large numbers of luteal cells compared to the control group. The prenatal PTU treated group did not exhibit a profound effect on ovarian morphology, histology, and morphometery. No difference was found in the serum estradiol concentration of control and PTU treated groups. J. Exp. Zool. 293:407–413, 2002. © 2002 Wiley‐Liss, Inc.
Conservation and variation in enamel protein distribution during vertebrate tooth development
Authors: Satchell Paul G. , Anderton Xochitl , Ryu Okhee H. , Luan Xianghong , Ortega Adam J. , Opamen Rene , Berman Brett J. , Witherspoon David E. , Gutmann James L. , Yamane Akira , Zeichner‐David Margerita , Simmer James P. , Shuler Charles F. , Diekwisch Thomas G.H.
Published: October 2002

Vertebrate enamel formation is a unique synthesis of the function of highly specialized enamel proteins and their effect on the growth and organization of apatite crystals. Among tetrapods, the physical structure of enamel is highly conserved, while there is a greater variety of enameloid tooth coverings in fish. In the present study, we postulated that in enamel microstructures of similar organization, the principle components of the enamel protein matrix would have to be highly conserved. In order to identify the enamel proteins that might be most highly conserved and thus potentially most essential to the process of mammalian enamel formation, we used immunoscreening with enamel protein antibodies as a means to assay for degrees of homology to mammalian enamel proteins. Enamel preparations from mouse, gecko, frog, lungfish, and shark were screened with mammalian enamel protein antibodies, including amelogenin, enamelin, tuftelin, MMP20, and EMSP1. Our results demonstrated that amelogenin was the most highly conserved enamel protein associated with the enamel organ, enamelin featured a distinct presence in shark enameloid but was also present in the enamel organ of other species, while the other enamel proteins, tuftelin, MMP20, and EMSP1, were detected in both in the enamel organ and in other tissues of all species investigated. We thus conclude that the investigated enamel proteins, amelogenin, enamelin, tuftelin, MMP20, and EMSP1, were highly conserved in a variety of vertebrate species. We speculate that there might be a unique correlation between amelogenin‐rich tetrapod and lungfish enamel with long and parallel crystals and enamelin‐rich basal vertebrate enameloid with diverse patterns of crystal organization. J. Exp. Zool. (Mol. Dev. Evol.) 294:91–106, 2002. © 2002 Wiley‐Liss, Inc.
Pentadactyl ground state of the avian wing
Authors: Larsson Hans C. E. , Wagner GÜnter P.
Published: October 2002

The issue of the homology of bird fingers with those of pentadactyl amniotes has been a topic of contention for nearly 200 years. Data from the fossil record and phylogenetic systematics ascribe bird digit homologies to digits I, II, and III of pentadactyl amniotes while embryological evidence supports digital homologies of II, III, and IV. Using a molecular marker specific for condensation competent mesenchymal cells, we describe a pentadactyl arrangement of prechondrogenic digital anlagen in the wings of stage 29 chick embryos. Only the middle three anlagen develop into mature fingers. This pattern supports the hypothesis that bird fingers develop from digital anlagen II, III, and IV of pentadactylous amniotes. In addition, this result rejects a model assuming a shift in the primary axis in bird digit development and shows that a prechondrogenic digital anlage has been maintained in the bird lineage for at least 220 million years since the last known pentadactylous ancestor of the lineage. Such a vestige suggests that strong constraints are maintaining a pentadactyl ground state in amniotes. J. Exp. Zool. (Mol. Dev. Evol.) 294:146–151, 2002. © 2002 Wiley‐Liss, Inc.
Circular chromatic numbers of some reduced Kneser graphs
Authors: Lih Ko‐Wei , Liu Daphne Der‐Fen
Published: October 2002

The vertex set of the reduced Kneser graph KG2(m,2) consists of all pairs {a,b} such that a, bε{1,2,…,m} and 2≤|a−b|≤m−2. Two vertices are defined to be adjacent if they are disjoint. We prove that, if m≥4 and m≠5, then the circular chromatic number of KG2(m,2) is equal to m−2, its ordinary chromatic number. © 2002 Wiley Periodicals, Inc. J Graph Theory 41: 62–68, 2002
Social funds: an effective instrument to support local action for poverty reduction?
Authors: de Haan Arjan , Holland Jeremy , Kanji Nazneen
Published: October 2002

This paper discusses whether social funds can help to support local action for poverty reduction. It focuses not on the specific poverty‐related projects that are supported by social funds, but the way social funds channel resources. In this context, this paper emphasizes the importance—and limitations—of the underlying poverty analysis informing the design of social funds, particularly the identification of beneficiaries. The paper describes the organizational characteristics social funds help to set up, and the relevance of decentralized organizations for community participation. The concluding section focuses on the implications of the tendency of social funds to create parallel structures. Copyright © 2002 John Wiley & Sons, Ltd.
Higher prevalence of anti‐HCV antibodies among HIV‐positive compared to HIV‐negative inhabitants of Addis Ababa, Ethiopia
Authors: Ayele Workenesh , Nokes D. James , Abebe Almaz , Messele Tsehaynesh , Dejene Amare , Enquselassie Fikre , Rinke de Wit Tobias F. , Fontanet Arnaud L.
Published: October 2002

Serum samples (n = 4,593) collected in 1994 as part of a representative household community survey of the population of Addis Ababa who were 0–49 years old were tested for hepatitis C (HCV) antibodies. A third generation ELISA was used for primary screening and a line immunoblot assay for confirmation. HCV antibody prevalence was 0.9% (95% CI, 0.6–1.2%) and higher among HIV‐positive compared to HIV‐negative individuals (4.5% vs. 0.8%, respectively, P < 0.001). Similar higher prevalence of HCV antibodies was seen among HIV‐positive compared to HIV‐negative antenatal care attenders (2.9% vs. 0.8%, respectively, P = 0.003, n = 1725), and sex workers (5.3% vs. 1.3%, respectively, P = 0.02, n = 383). Such association between HCV and HIV infection has not been described previously in Africa. After stratification by HIV status, HCV prevalence among women of the general population was identical to that of sex workers, suggesting that HCV sexual transmission is not common in this population and that HIV infection does not enhance susceptibility to HCV sexual transmission. J. Med. Virol. 68:12–17, 2002. © 2002 Wiley‐Liss, Inc.
Equivalent circuit representation for integral formulations of electromagnetic problems
Authors: Baudrand H. , Bajon D.
Published: October 2002

The purpose of this study is to investigate the resources offered by electromagnetic equivalent circuits as a tool for the systematic derivation of integral equations and also, to propose consistent models for active sources. Copyright © 2002 John Wiley & Sons, Ltd.
Parameters estimation of the d.c. electrothermal model of the bipolar transistor
Authors: Zarębski Janusz , Górecki Krzysztof
Published: October 2002

This paper concerns the problems of the parameter values estimation of d.c. electrothermal model of the BJT formulated for circuit analysis with SPICE. In this case, PARTS software available in SPICE cannot be used. In the paper, a new estimation algorithm for d.c. electrothermal model of the BJT is proposed. The form of the electrothermal BJT model is also presented. The estimation algorithm implemented into the computer‐controlled measurement set allows one to derive the values of the parameters automatically after the measurements of the selected isothermal characteristics and proper calculations. Copyright © 2002 John Wiley & Sons, Ltd.
Estrogen treatment suppresses forebrain p75 neurotrophin receptor expression in aged, noncycling female rats
Authors: Ping Sophie E. , Greferath Ursula , Barrett Graham L.
Published: October 2002

There is increasing evidence that estrogen has beneficial effects on cognition, both in humans and in rodents, and may delay Alzheimer's disease onset in postmenopausal women. Several rodent studies have utilised the ovariectomy model to show estrogen regulation of the p75 neurotrophin receptor, TrkA, and markers of acetylcholine synthesis in the cholinergic basal forebrain. We studied estrogenic effects in aged (16–17‐month‐old), noncycling rats. Estrogen treatment for 10 days drastically reduced p75NTR immunoreactivity in the rostral parts of the basal forebrain. The number of p75NTR‐immunoreactive neurons was decreased, and those neurons remaining positive for p75NTR showed reduced p75NTR staining intensity. In vehicle‐treated rats, almost all choline acetyltransferase‐immunoreactive neurons were p75NTR positive (and vice versa), but, in estrogen treated rats, large numbers of choline acetyltransferase‐immunoreactive cells were negative for p75NTR. Similar levels of p75NTR down‐regulation in the rostral basal forebrain were found when estrogen treatment was extended to 6 weeks. There was no reduction in the number of p75NTR‐immunoreactive neurons in the caudal basal forebrain after 10 days of treatment. After 6 weeks of treatment, however, there was evidence of p75NTR down‐regulation in the caudal basal forebrain. There was no evidence of hypertrophy or atrophy of cholinergic neurons even after 6 weeks of estrogen treatment. Considering the evidence for the role of p75NTR in regulating survival, growth and nerve growth factor responsiveness of cholinergic basal forebrain neurons, the results indicate an important aspect of estrogen's effects on the nervous system. © 2002 Wiley‐Liss, Inc.
Molecular mechanisms involved in the actions of apotransferrin upon the central nervous system: Role of the cytoskeleton and of second messengers
Authors: Marta Cecilia B. , Davio Carlos , Pasquini Laura A. , Soto Eduardo F. , Pasquini Juana M.
Published: October 2002

Apotransferrin (aTf), intracranially administered into newborn rats, produces increased myelination with marked increases in the levels of myelin basic protein (MBP), phospholipids and galactolipids, and mRNAs of MBP and 2′, 3′ cyclic nucleotide 3′‐phosphohydrolase (CNPase). Cytoskeletal proteins such as tubulin, actin, and microtubule‐associated proteins are also increased after aTf injection. In contrast, almost no changes are observed in myelin proteolipid protein (PLP) or in its mRNA or cholesterol. In the present study, we used brain‐tissue slices and cell cultures highly enriched for oligodendroglia to investigate signaling pathways involved in the action of aTf, and to find out whether cytoskeletal integrity and dynamics were essential for its action upon the neural expression of certain genes. Treatment of brain‐tissue slices with aTf produced a marked increase in the expression of MBP, CNPase, and tubulin mRNAs. Colchicine, cytochalasin, and taxol severely reduced the effect of aTf. Addition to cultures of an antibody against transferrin receptor (TfR), protein kinase inhibitors, or a cyclic AMP (cAMP) analogue showed that a functionally intact TfR was necessary, and that tyrosine kinase, protein kinase C and A, as well as calcium‐calmodulin‐dependent kinase (Ca‐CaMK) activities appeared to mediate aTf actions upon the expression of the above mentioned genes. Changes in the levels of phosphoinositides and cAMP induced by aTf in oligodendroglial cell (OLGc) cultures correlated with these results and coincide with an activation of the cyclic response element binding protein (CREB) and of mitogen activated protein kinases. The increased expression of certain myelin genes produced by aTf appear to be mediated by interaction of this glycoprotein with its receptor, by the cytoskeleton of the OLGc, and by a complex activation of protein kinases which lead to CREB phosphorylation. © 2002 Wiley‐Liss, Inc.
Chronology of the last recession of the Greenland Ice Sheet
Authors: Bennike Ole , Björck Svante
Published: October 2002

A new deglaciation chronology for the ice‐free parts of Greenland, the continental shelf and eastern Ellesmere Island (Canada) is proposed. The chronology is based on a new compilation of all published radiocarbon dates from Greenland, and includes crucial new material from southern, northeastern and northwestern Greenland. Although each date provides only a minimum age for the local deglaciation, some of the dates come from species that indicate ice‐proximal glaciomarine conditions, and thus may be connected with the actual ice recession. In addition to shell dates, dates from marine algae, lake sediments, peat, terrestrial plants and driftwood also are included. Only offshore and in the far south have secure late‐glacial sediments been found. Other previous reports of late‐glacial sediments (older than 11.5 cal. kyr BP) from onshore parts of Greenland need to be confirmed. Most of the present ice‐free parts of Greenland and Nares Strait between Greenland and Ellesmere Island were not deglaciated until the early Holocene. Copyright © 2002 John Wiley & Sons, Ltd.
An enzyme‐based method for the rapid determination of sucrose, glucose and fructose in sugar beet roots and the effects of impact damage and postharvest storage in clamps
Authors: Spackman Victoria M T , Cobb Andrew H
Published: October 2002

A high‐throughput enzyme‐coupled assay is described for the determination of sucrose, glucose and fructose in sugar beet roots. This method is sensitive, rapid and inexpensive and has been used to highlight the increases in sucrose loss following root stresses such as freezing or aggressive harvesting. Sugar beet roots lose 12.5% of their sucrose following an episode of impact damage greater than 2 J, rising to 19.7% loss after 8 J, with concurrent increases in glucose and fructose. Increases in glucose and fructose are particularly pronounced following a period of clamp storage (up to 2.3 and 3.3 µg mg−1 fresh weight, respectively).

© 2001 Society of Chemical Industry
Effect of different organic growing conditions on quality of cooked potatoes
Authors: Thybo Anette K , Mølgaard Jens Peter , Kidmose Ulla
Published: October 2002

In 1997 and 1998 the effect of six different organic treatments on the chemical, rheological and sensory quality of cooked potatoes was investigated. Cattle slurry and cattle deep litter were each applied to sandy soil in three ways. The type of manure applied gave the most distinct effect on sensory quality. Manuring with slurry decreased dry matter, starch and phytic acid and increased N and rheological properties (modulus of deformability, stress, strain) significantly compared with deep litter manuring. Maturity of tubers was assumed to be the main cause of these differences. Regarding the sensory quality of potatoes, minor differences were found among the organic treatments investigated. Potatoes manured with slurry were slightly moister and seemed to have higher discolouration and off‐odour and lower yellowness than potatoes manured with deep litter. Mealiness was highest when composted deep litter was ploughed in and lowest when deep litter and straw were ploughed in. The observed differences in sensory quality are presumably an effect of availability of N in the manure. Chlorogenic acid was the only phenolic acid that was affected by the organic treatments. The variation in phenolic acid and scopoletin contents did not seem to influence the off‐taste of potatoes. As the sensory quality of potatoes produced by different organic treatments had not been addressed previously, this study revealed interesting results.

© 2001 Society of Chemical Industry
Atypical antibody specificity: advancing the development of a generic assay for sulphonamides using heterologous ELISA
Authors: Spinks C A , Wyatt G M , Everest S , Jackman R , Morgan M R A
Published: October 2002

Development of immunoassays with broad specificity recognition of the sulphonamide drugs has proved surprisingly difficult in previous work. We have taken an antiserum specific for sulphachlorpyridazine in an ELISA system homologous for immunogen and solid phase conjugate and used it in a heterologous ELISA format. The assay proved to have broad specificity, recognising 15 out of 21 drugs tested. Despite low cross‐reactivity (<1%) for seven of the drugs tested, a fully optimised assay could prove useful as a multiresidue method for the group of drugs. The results provide evidence regarding the binding characteristics of the antibody and the immunogenicity of the target.

© 2002 Society of Chemical Industry
Wet separation of wheat flours into starch and gluten fractions: the combined effects of water to flour ratio–dough maturation time and the effects of flour aging and ascorbic acid addition
Authors: Dik Tunay , Yöndem‐Makascıoğlu Füsun , Aytaç Cem H , Kıncal N Suzan
Published: October 2002

The process of making a dough, allowing time for maturation, dispersing the dough in water and wet sieving/washing to obtain a protein fraction and starch milk was studied using response surface methodology by changing the water to flour ratio in dough making (400–710 g kg−1), maturation time (45–660 s) and the type of flour. Two grades of bread wheat flour and durum clear flour were studied. The effects of aging at ambient temperature for up to 29 days and the addition of ascorbic acid at 100 or 500 mg kg−1 flour on separation behaviour were also studied for freshly milled high‐grade (65% extraction) bread wheat flours at constant maturation time, 600 s, and at optimum farinograph water absorption value. The quantities and dry matter contents of the protein fraction and starch milk were measured; a sample of starch milk was centrifuged to obtain decantate, tailings and prime starch fractions, and the dry matter contents of each were determined. All the dried samples were also analysed for protein content. The fractional recoveries of dry matter and protein in the protein fraction, prime starch, tailings and decantate were calculated for each experiment. The acid values of flour oils were also determined on some aged flour samples. The results indicated superior separation characteristics in high‐grade wheat flour compared with lower‐grade flours. The water to flour ratio was more influential than maturation time within the range studied. Contrary to the initial expectation, no statistically significant effect of flour aging was observed in the studies with no additive, and ascorbic acid addition was not found to improve the wet separation behaviour, the separation behaviour becoming even worse at the 100 mg kg−1 level. Acid value showed a slight increase with time.

© 2002 Society of Chemical Industry