bluuebunny/crossref_metadata_2025_split huggingface dataset:
Small Guanosine Triphospatase RhoA and Rho-Associated Kinase as Regulators of Trophoblast Migration
Authors: Shiokawa Shigetatsu , Iwashita Mitsutoshi , Akimoto Yoshihiro , Nagamatsu Shinya , Sakai Ken , Hanashi Hideki , Kabir-Salmani Maryam , Nakamura Yukio , Uehata Masayoshi , Yoshimura Yasunori 
Published: December 2002

AbstractThe small guanosine triphosphatase Rho controls cell adhesion and motility through reorganization of the actin cyto-skeleton and regulation of actomyosin contractility. Among the putative target molecules of Rho, a Rho-associated coiled coil-forming protein kinase (ROCK) is thought to participate in Rho-mediated cell adhesion and motility. In the present study, we explored the expression and function of RhoA and ROCK in human trophoblast cells. The colocalization of RhoA, cytokeratin 8/18, and cytokeratin 7 in some cells located in the decidual stromal region indicated that extravillous trophoblast cells expressed RhoA. In double staining for RhoA and ROCK in human chorionic villi, RhoA staining was strongly positive in the cytoplasm of cytotrophoblasts, whereas ROCK stained in the cytoplasm of cytotrophoblasts and syncytiotrophoblasts. Both RhoA and ROCK were stained in cytoplasma of cultured human cytotrophoblast. Cultured human trophoblast cells contained actin stress fibers that were lost after treatment with C3, an exoenzyme produced by Clostridium botulinum. Y-27632, a selective ROCK inhibitor, suppressed RhoA-induced formation of actin stress fibers and formation of focal contact in trophoblast cells. The trophoblast reacquired actin stress fibers and focal contact after withdrawal of Y-27632. Cultured human cytotrophoblast cells from 7–9 wk of gestation migrated into a fibronectin-coated membrane. Both C3 exoenzyme and Y-27632 inhibited cytotrophoblast migration in a dose-dependent manner. In conclusion, cyto-trophoblasts express RhoA and ROCK in their cytoplasm, and RhoA-ROCK is involved in their assembly of actin stress fibers. Suppression of RhoA-ROCK reduces trophoblast migration. These findings suggest that RhoA-ROCK signaling is a key regulator of trophoblast cell migration.
Characterization of Morphological and Cytoskeletal Changes in Trophoblast Cells Induced by Insulin-Like Growth Factor-I
Authors: Kabir-Salmani Maryam , Shiokawa Shigetatsu , Akimoto Yoshihiro , Hasan-Nejad Habib , Sakai Keiji , Nagamatsu Shinya , Sakai Ken , Nakamura Yukio , Hosseini Ahmad , Iwashita Mitsutoshi 
Published: December 2002

AbstractIGF-I and IGF-II were appeared to play major roles in the adhesive and migratory events that are considered to be crucial in the implantation process. The purpose of this study was to determine the effects of IGF-I on trophoblast adhesion to extracellular matrix. Trophoblast cells obtained from early gestation at artificial abortion were incubated with the indicated doses of IGF-I at the indicated times. Trophoblast cells were treated with IGF-I in the presence or absence of RGD peptide and an antibody against α-subunit of IGF-I receptor (αIR3). Morphometric and morphological changes were studied using light and electron microscopy. Furthermore, vinculin, actin stress fibers, phosphorylated focal adhesion kinase (FAK), phosphotyrosine, and paxillin were immunolocalized in trophoblast cells after IGF-I treatment in the presence or absence of αIR3. Immunoprecipitation and anti-phosphotyrosine immunoblotting were carried out to detect the phosphorylated FAK and phosphorylated paxillin contents of the IGF-I-treated and untreated trophoblast cells. The results showed that IGF-I promoted trophoblast adhesion to fibronectin substrate in a time- and dose-dependent manner, and addition of RGD peptide and αIR3 monoclonal antibody abolished the effects of IGF-I in these cells. Morphological studies exhibited an increase in the lamellipodia formation upon IGF-I treatment, and confocal images of immunofluorescent staining revealed localization of phosphorylated FAK, paxillin, and vinculin at focal adhesions as well as redistribution of actin microfilaments and formation of actin stress fibers inside the cell. Western blotting, using antiphosphotyrosine demonstrated proteins with molecular masses of 125 kDa (FAK) and 68 kDa (paxillin) present in the IGF-I-treated cells, which were lacking in the control groups. In conclusion, these findings suggest that IGF-I can stimulate lamellipodia formation and promote adhesion of trophoblast cells to extracellular matrix by activating their adhesion molecules that must be activated within the implantation window.
Systemic Distribution of Steroid Sulfatase and Estrogen Sulfotransferase in Human Adult and Fetal Tissues
Authors: Miki Yasuhiro , Nakata Taisuke , Suzuki Takashi , Darnel Andrew David , Moriya Takuya , Kaneko Chika , Hidaka Kumiko , Shiotsu Yukimasa , Kusaka Hideaki , Sasano Hironobu 
Published: December 2002

Abstract
Estrogens play a key role in various target tissues. Enzymes involved in the biosynthesis and metabolism of these sex steroids also regulate estrogenic actions in these tissues. Estrone sulfate (E1S) is a major circulating plasma estrogen that is converted into the biologically active estrogen, estrone (E1), by steroid sulfatase (STS). E1 is also sulfated and reverted into E1S by estrogen sulfotransferase (EST). These two enzymes have recently been shown to play important roles in the in situ estrogen actions of various sex steroid-dependent human tumors. However, the distribution of STS and EST in normal adult and fetal human tissues remains largely unknown. Therefore, in this study, in addition to examining the tissue distribution of both STS and EST mRNA in human adult and fetal tissues using RT followed by quantitative PCR, we studied the activity of these enzymes using 3H-labeled E1/E1S as substrates in the homogenates of various human adult tissues. We also examined the localization of STS and EST protein in human adult and fetal tissues using immunohistochemistry, and that of EST mRNA in the adult kidney using laser dissection microscopy and PCR. STS mRNA, enzyme activity, and immunoreactivity were either absent or detected at very low levels in all adult and fetal tissues examined in this study. EST mRNA expression, however, was detected in all of the tissues examined, except for adult spleen and pancreas. EST enzyme activities were consistent with those of mRNA expression in the great majority of the tissues examined. Marked EST immunoreactivity was detected in hepatocytes, adrenal gland (adult, zona fasciculate to the reticularis; fetus, fetal zone), and epithelial cells of the gastrointestinal tract, smooth muscle cells of the tunica media in aorta, Leydig cells of the testis, and syncytiotrophoblast of the placenta. Patterns of EST immunolocalization were similar between adult and fetal human tissues, but EST immunoreactivity was detected in the urinary tubules of adult kidney, whereas in the fetal kidney, it was localized in the interstitial cells surrounding the urinary tubules. In the adult kidney, the presence of EST mRNA was also confirmed in the cells of urinary tubules using laser dissection microscopy and RT-PCR.
Although the number of human tissues available for examination in this study was limited, our results suggest that between the enzymes involved in estrogen activation or inactivation, EST and not STS is the more widely expressed enzyme in various peripheral tissues in humans. We speculate that EST may play an important role in protecting peripheral tissues from possible excessive estrogenic effects.
Thyroid Acropachy: Report of 40 Patients Treated at a Single Institution in a 26-Year Period
Authors: Fatourechi Vahab , Ahmed Debra D. F. , Schwartz Kara M. 
Published: December 2002

AbstractThyroid acropachy is an extreme manifestation of autoimmune thyroid disease. It presents with digital clubbing, swelling of digits and toes, and periosteal reaction of extremity bones. It is almost always associated with ophthalmopathy and thyroid dermopathy. During a 26-yr period at our institution, of 178 patients with thyroid dermopathy, 40 had acropachy. Clubbing associated with thyroid dermopathy (pretibial myxedema) was seen in 35 patients. Clubbing usually was not a patient complaint and was noted only by clinical observers. Four of eight patients with hand and extremity radiographs had periosteal reaction. Seven had associated extremity and joint pain; this pain was absent at long-term follow-up. Half of the patients required systemic corticosteroid therapy, 53% required transantral or transfrontal orbital decompression for severe ophthalmopathy, and 18% had the elephantiasic form of dermopathy. Cigarette-smoking rates were 81% for women and 75% for men (mean, 28 pack-years). All 13 patients who had thyroid-stimulating Ig measurement had high titers. Long-term follow-up (median, 12.5 yr) revealed that acropachy was not a complaint in follow-up visits or questionnaires. The data suggest that thyroid acropachy is an indicator of severity of ophthalmopathy and dermopathy. It is a source of clinical concern only if dermopathy is persistent and severe.
Growth Hormone Blunts Protein Oxidation and Promotes Protein Turnover to a Similar Extent in Abdominally Obese and Normal-Weight Women
Authors: Buijs Madelon M. , Romijn Johannes A. , Burggraaf Jacobus , De Kam Marieke L. , Cohen Adam F. , Frölich Marijke , Stellaard Frans , Meinders A. Edo , Pijl Hanno 
Published: December 2002

Abstract
Abdominally obese individuals have reduced 24-h plasma GH concentrations. Their normal plasma IGF-I levels may reflect GH hypersensitivity. Alternatively, obesity-associated hyposomatotropism may cause less biological effect in target tissues. We therefore determined whole-body responsiveness to the anabolic effects of GH in abdominally obese (OB) and normal weight (NW) premenopausal women. A 1-h iv infusion of GH or placebo was randomly administered to six NW (body mass index, 21.1 ± 1.9 kg/m2) and six OB (body mass index, 35.5 ± 1.5 kg/m2) women in a cross-over design. Endogenous insulin, glucagon and GH secretion was suppressed by infusion of somatostatin. Whole-body protein turnover was measured using a 10-h infusion of [13C]-leucine. GH administration induced a similar plasma GH peak in NW and OB women (49.8 ± 10.4 vs. 45.1 ± 5.6 mU/liter). GH, compared with placebo infusion, increased nonoxidative leucine disposal, P &lt; 0.0001) and endogenous leucine appearance (Ra, P = 0.0004) but decreased leucine oxidation (P = 0.0051). All changes were similar in both groups. Accordingly, whole-body GH responsiveness, defined as the maximum response of nonoxidative leucine disposal, leucine Ra, and oxidation per unit of GH, was not different in OB and NW women (0.25 ± 0.18 vs. 0.19 ± 0.17 μmol/kg·h, 0.21 ± 0.23 vs. 0.13 ± 0.17 μmol/kg·h, and −0.10 ± 0.08 vs. −0.08 ± 0.05 μmol/kg·h, respectively). These results indicated that whole-body tissue responsiveness to the net anabolic effect of GH is similar in OB and NW women. Hence, we inferred that hyposomatotropism may promote amino acid oxidation and blunt protein turnover in abdominal obesity. However, hyposomatotropism cannot account for all anomalous features of protein metabolism in abdominally obese humans.
Demonstration of Enhanced Potency of a Chimeric Somatostatin-Dopamine Molecule, BIM-23A387, in Suppressing Growth Hormone and Prolactin Secretion from Human Pituitary Somatotroph Adenoma Cells
Authors: Saveanu A. , Lavaque E. , Gunz G. , Barlier A. , Kim S. , Taylor J. E. , Culler M. D. , Enjalbert A. , Jaquet P. 
Published: December 2002

Abstract
In acromegaly, the combination of somatostatin (SS) and dopamine (DA) agonists has been shown to enhance suppression of GH secretion. In the present study, a new chimeric molecule, BIM-23A387, which selectively binds to the SS subtype 2 receptor (sst2; Ki = 0.10 nm) and to the DA D2 receptor (D2DR; Ki = 22.1 nm) was tested in cultures prepared from 11 human GH-secreting tumors for its ability to suppress GH and prolactin (PRL) secretion. The chimeric compound was compared with individual sst2 and D2DR agonists of comparable activity at the individual receptors. All tumors expressed both sst2 and D2DR mRNAs (0.8 ± 0.2 and 4.7 ± 0.7 copy/copy β-glucuronidase mRNA, respectively). In cell cultures from seven octreotide-sensitive tumors, the maximal inhibition of GH release induced by the individual sst2 and D2DR analogs and by BIM-23A387 was similar. However, the mean EC50 for GH suppression by BIM-23A387 (0.2 pm) was 50 times lower than that of the individual sst2 and D2DR analogs, either used individually or combined. Similar data were obtained in four tumors that were only partially responsive to octreotide. The inhibition of GH release by BIM-23A387 was only partially reversed by the D2R2 antagonist, sulpiride, or by the sst2 antagonist, BIM-23454. Only when both antagonists were combined was the GH suppressive effect of BIM-23A387 totally reversed. Finally, BIM-23A387 produced a mean 73 ± 6% inhibition of PRL in six mixed GH plus PRL tumors. These data demonstrate an enhanced potency of the chimeric molecule, BIM-23A387, in suppressing GH and PRL secretion from acromegalic tumors, which cannot be explained merely on the basis of binding affinity for SS and/or DA receptors.
Effects of Pioglitazone in Nondiabetic Patients with Arterial Hypertension: A Double-Blind, Placebo-Controlled Study
Authors: Füllert S. , Schneider F. , Haak E. , Rau H. , Badenhoop K. , Lübben G. , Usadel K.-H. , Konrad T. 
Published: December 2002

Abstract
Hypertension is often associated with insulin resistance, dyslipidemia and obesity, which indicate a prediabetic state and increased risk of cardiovascular disease. Pioglitazone treatment of patients with type 2 diabetes reduces insulin resistance and improves lipid profiles. The present double-blind placebo-controlled study is the first study to report effects of pioglitazone in non-diabetic patients with arterial hypertension. Following a one week run-in, 60 patients were randomized to receive either pioglitazone (45 mg/day) or placebo for 16 weeks. Insulin sensitivity (M-value) increased by 1.2 ± 1.7 mg/min/kg with pioglitazone compared with 0.4 ± 1.4 mg/min/kg (P = 0.022) with placebo. HOMA index was decreased (−22.5 ± 45.8) by pioglitazone but not by placebo (+0.8 ± 26.5; P &lt; 0.001). Decreases in fasting insulin and glucose were significantly (P = 0.002 and P = 0.004, respectively) greater with pioglitazone than placebo. Body weight did not change significantly with either treatment. HDL-cholesterol was increased and apolipoprotein B was decreased to a significantly greater extent with pioglitazone. There was a significantly (P = 0.016) greater decrease from baseline in diastolic blood pressure with pioglitazone. These changes would suggest improved glucose metabolism and a possible reduction in risk of cardiovascular disease with pioglitazone treatment of non-diabetic patients with arterial hypertension.
11β-Hydroxysteroid Dehydrogenase Types 1 and 2: An Important Pharmacokinetic Determinant for the Activity of Synthetic Mineralo- and Glucocorticoids
Authors: Diederich Sven , Eigendorff Ekkehard , Burkhardt Patrick , Quinkler Marcus , Bumke-Vogt Christiane , Rochel Marina , Seidelmann Dieter , Esperling Peter , Oelkers Wolfgang , Bähr Volker 
Published: December 2002

AbstractThe 11β-hydroxysteroid dehydrogenase (11β-HSD) system plays a pivotal role in glucocorticoid (GC) and mineralocorticoid (MC) action. Although 11β-HSD activities are important determinants for the efficacy of synthetic MCs and GCs, corresponding pharmacokinetic data are scanty. Therefore, we characterized 11β-HSD profiles for a wide range of steroids often used in clinical practice. 11β-HSD1 and 11β-HSD2 were selectively examined in 1) human liver and kidney cortex microsomes, and 2) Chinese hamster ovarian cells stably transfected with 11β-HSD1 or 11β-HSD2 expression vectors. Both systems produced concordant evidence for the following conclusions.Oxidation of steroids by 11β-HSD2 is diminished if they are fluorinated in position 6α or 9α (e.g. in dexamethasone) or methylated at 2α or 6α (in methylprednisolone) or 16α or 16β, by a methylene group at 16 (in prednylidene), methyloxazoline at 16, 17 (in deflazacort), or a 2-chlor configuration. Whereas the methyl groups also decrease reductase activity (steric effects), fluorination increases reductase activity (negative inductive effect), leading to a shift to reductase activity. This may explain the strong MC activity of 9α-fluorocortisol and should be considered in GC therapy directed to 11β-HSD2-expressing tissues (kidney, colon, and placentofetal unit). 11β-HSD2 oxidation of prednisolone is more effective than that of cortisol, explaining the reduced MC activity of prednisolone compared with cortisol.Reduction by 11β-HSD1 is diminished by 16α-methyl, 16β-methyl, 2α-methyl, and 2-chlor substitution, whereas it is increased by the Δ1-dehydro configuration in prednisone, resulting in higher hepatic first pass activation of prednisone compared with cortisone.To characterize a GC or a MC as substrate for the different 11βHSDs may be essential for an optimized steroid therapy.
Control of Texture in ZnO by Slip Casting in a Strong Magnetic Field and Heating
Authors: Suzuki Tohru S , Sakka Yoshio 
Published: December 2002

Abstract
Anisotropic susceptibilities of feeble magnetic ceramics such as zinc oxide are quite small; therefore, it had been difficult to control the textured microstructures of these ceramics using a magnetic field. However, fabrication of the textured ZnO is achieved by slip casting in a strong magnetic field followed by heating. After the stable suspension of ZnO fine particles was compacted by slip casting in a strong magnetic field, a green body with a slight degree of crystalline texture was obtained. Densification and the grain growth enhanced the degree of texture during the sintering.
Substrate and product inhibition of hydrogen production by the extreme thermophile, Caldicellulosiruptor saccharolyticus
Authors: van Niel Ed W. J. , Claassen Pieternel A. M. , Stams Alfons J. M. 
Published: December 2002

AbstractSubstrate and product inhibition of hydrogen production during sucrose fermentation by the extremely thermophilic bacterium Caldicellulosiruptor saccharolyticus was studied. The inhibition kinetics were analyzed with a noncompetitive, nonlinear inhibition model. Hydrogen was the most severe inhibitor when allowed to accumulate in the culture. Concentrations of 5–10 mM H2 in the gas phase (≡ partial hydrogen pressure (pH2) of (1–2) · 104 Pa) initiated a metabolic shift to lactate formation. The extent of inhibition by hydrogen was dependent on the density of the culture. The highest tolerance for hydrogen was found at low volumetric hydrogen production rates, as occurred in cultures with low cell densities. Under those conditions the critical hydrogen concentration in the gas phase was 27.7 mM H2 (≡ pH2 of 5.6 · 104 Pa); above this value hydrogen production ceased completely. With an efficient removal of hydrogen sucrose fermentation was mainly inhibited by sodium acetate. The critical concentrations of sucrose and acetate, at which growth and hydrogen production was completely inhibited (at neutral pH and 70°C), were 292 and 365 mM, respectively. Inorganic salts, such as sodium chloride, mimicked the effect of sodium acetate, implying that ionic strength was responsible for inhibition. Undissociated acetate did not contribute to inhibition of cultures at neutral or slightly acidic pH. Exposure of exponentially growing cultures to concentrations of sodium acetate or sodium chloride higher than ca. 175 mM caused cell lysis, probably due to activation of autolysins. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 81: 255–262, 2003.
Effects of methanol concentration on expression levels of recombinant protein in fed‐batch cultures of Pichia methanolica
Authors: Mayson Brian E. , Kilburn Douglas G. , Zamost Bruce L. , Raymond Christopher K. , Lesnicki Gary J. 
Published: December 2002

AbstractThe methylotrophic yeast Pichia methanolica can be used to express recombinant genes at high levels under the control of the methanol‐inducible alcohol oxidase (AUG1) promoter. Methanol concentrations during the induction phase directly affect cellular growth and protein yield. Various methanol concentrations controlled by an on‐line monitoring and control system were investigated in mixed glucose/methanol fed‐batch cultures of P. methanolica expressing the human transferrin N‐lobe protein. The PMAD18 P. methanolica strain utilized is a knock‐out for the chromosomal AUG1 gene locus, resulting in a slow methanol utilization phenotype. Maximum growth of 100 g of dry cell weight per liter of culture was observed in cultures grown at 1.0% (v/v) methanol concentration. Maximum recombinant gene expression was observed for cultures controlled at 0.7% (v/v) methanol concentration, resulting in maximum volumetric production of 450 mg of transferrin per liter after 72 h of elapsed fermentation time. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 81: 291–298, 2003.
Kinetic analysis of bacterial bioluminescence
Authors: Kelly Christine J. , Hsiung Chia‐Joung , Lajoie Curtis A. 
Published: December 2002

AbstractBioluminescence from the lux‐based bacterial reporter Pseudomonas fluorescens HK44 was experimentally investigated under growth substrate‐rich and limiting conditions in batch, continuous stirred tank (CSTR), and turbidostat reactors. A mechanistically based, mathematical model was developed to describe bioluminescence based on 1) production and decay of catalytic enzymes, and 2) reactant cofactor availability. In the model, bioluminescence was a function of inducer, growth substrate, and biomass concentration. A saturational dependence on growth substrate concentration accommodated dependence on cofactor availability and inducer concentration to accommodate enzyme production was incorporated in the model. Under growth substrate and inducer limiting conditions in the batch reactor and CSTR, bioluminescence was found to decrease in response to cellular energy limitations. The effective lux system enzyme decay rate was determined in independent measurements to be 0.35 hr−1 and the model captured most of the bioluminescent behavior, except at long growth times and high cell density. © 2003 Wiley Periodicals, Inc. Biotechnol Bioeng 81: 370–378, 2003.