Artykuły w czasopismach na temat „F-cell ratio”
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Choi, JW, Y. Kim, M. Fujino i M. Ito. "Significance of fetal hemoglobin-containing erythroblasts (F blasts) and the F blast/F cell ratio in myelodysplastic syndromes". Leukemia 16, nr 8 (31.07.2002): 1478–83. http://dx.doi.org/10.1038/sj.leu.2402536.
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Haller, M., H. Brechtelsbauer, C. Akbulut, W. Fett, J. Briegel i U. Finsterer. "Isovolemic Hemodilution Alters the Ratio of Whole-Body to Large-Vessel Hematocrit (F-Cell Ratio)". Transfusion Medicine and Hemotherapy 22, nr 2 (1995): 74–80. http://dx.doi.org/10.1159/000223103.
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Isbister, J. P. "The F-Cell Ratio: A Clinically Important Parameter or Just Fine Tuning?" Transfusion Medicine and Hemotherapy 22, nr 2 (1995): 69–70. http://dx.doi.org/10.1159/000223101.
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Khan, Faraz A., Richard Mullins, Anna M. Ledgerwood i Charles E. Lucas. "Variability of the f-cell ratio after treatment of traumatic hemorrhagic shock". Annals of Medicine and Surgery 35 (listopad 2018): 176–79. http://dx.doi.org/10.1016/j.amsu.2018.10.001.
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Puty, Bruna, Leonardo Oliveira Bittencourt, Iago Cesar Nogueira, Marília Afonso Rabelo Buzalaf, Edivaldo Herculano Oliveira i Rafael Rodrigues Lima. "Human cultured IMR-32 neuronal-like and U87 glial-like cells have different patterns of toxicity under fluoride exposure". PLOS ONE 16, nr 6 (17.06.2021): e0251200. http://dx.doi.org/10.1371/journal.pone.0251200.
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Background Fluoride (F) is a naturally exists in nature but several studies have indicated it as an environmental toxicant to all leaving beings. Human F exposure has increased over the years since this ion has been used by industry on foods, beverages, toothpastes and on water supply. Although F is safe at optimal concentrations in water supply, human exposure to high levels could trigger neurofunctional deficits. Materials and methods In this study, human glial-like (U87) and neuronal-like (IMR-32) cells lineages were used to access F toxicity and CNS cell sensibility on both cell facing the same protocol. Cells were exposed to F over 3, 5 and 10 days on two different F concentrations. Fluoride exposed cells were evaluated by standard toxicity assays to cell viability, apoptosis, necrosis and general cell metabolism. Oxidative stress parameters were evaluated by ATP and ROS levels, lipid peroxidation, GSH/GSSG ratio and comet assay. Results No changes were observed in IMR-32 at any given time while after 10 days of exposure to 0.22μg/mL, U87 glial-like cells showed signs of toxicity such as decreased cell viability by necrosis while general cell metabolism was increased. Oxidative stress parameters were next evaluated only on U87 glial-like cells after 10 days of exposure. F induced a decrease on ATP levels while no changes were observed on reactive oxygen species and lipid peroxidation. GSH/GSSG ratio was decreased followed by DNA damage both on 0.22μg/mL F. Conclusions Our results suggest an important differential behavior of the distinct types of cells exposed to the different fluoride concentrations, pointing that the U87 glial-like cells as more susceptible to damage triggered by this ion.
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Chen, Jin-Shuen, Li-Chien Chang, Chia-Chao Wu, Lai-King Yeung i Yuh-Feng Lin. "Involvement of F-Actin in Chaperonin-Containing t-Complex 1 Beta Regulating Mouse Mesangial Cell Functions in a Glucose-Induction Cell Model". Experimental Diabetes Research 2011 (2011): 1–10. http://dx.doi.org/10.1155/2011/565647.
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The aim of this study is to investigate the role of chaperonin-containing t-complex polypeptide 1 beta (CCT2) in the regulation of mouse mesangial cell (mMC) contraction, proliferation, and migration with filamentous/globular-(F/G-) actin ratio under high glucose induction. A low CCT2 mMC model induced by treatment of small interference RNA was established. Groups with and without low CCT2 induction examined in normal and high (H) glucose conditions revealed the following major results: (1) low CCT2 or H glucose showed the ability to attenuate F/G-actin ratio; (2) groups with low F/G-actin ratio all showed less cell contraction; (3) suppression of CCT2 may reduce the proliferation and migration which were originally induced by H glucose. In conclusion, CCT2 can be used as a specific regulator for mMC contraction, proliferation, and migration affected by glucose, which mechanism may involve the alteration of F-actin, particularly for cell contraction.
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Dover, GJ, VT Chang, SH Boyer, GR Serjeant, S. Antonarakis i DR Higgs. "The cellular basis for different fetal hemoglobin levels among sickle cell individuals with two, three, and four alpha-globin genes". Blood 69, nr 1 (1.01.1987): 341–44. http://dx.doi.org/10.1182/blood.v69.1.341.341.
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Abstract Fetal hemoglobin (HbF) levels vary widely among individuals with sickle cell anemia (SS). Previous studies have suggested that HbF levels in SS individuals with alpha-thalassemia (two or three functional alpha- globin genes) are lower than HbF levels in SS individuals with four normal alpha-globin genes. Using immunocytochemical techniques, we studied F cell production as measured by % F reticulocytes, the amount of HbF per F cell, and the preferential survival of F cells versus non- F cells in 51 subjects with four alpha genes, 32 subjects with three alpha genes, and 18 subjects with two alpha genes. Comparison between alpha-globin gene groups was performed for the total sample as well as for a subset of 82 individuals who had replicate samples and a further subset of 39 age-matched individuals. %HbF levels were 6.8, 4.9, and 4.5 percent for the total four-, three-, and two-alpha-globin-gene groups, respectively. The percentage of F reticulocytes, percentage HbF per F cell, and the enrichment ratio (% F cell/% F reticulocytes) did not change significantly with alpha-globin gene number. Moreover, no correlation existed between alpha-globin gene number and the absolute number of F cells in any group studied. However, there was a strong inverse correlation (r = -0.407, P = .0001) between non-F cell levels (1.7 +/- 2, 2.2 +/- 5, 3.0 +/- 1.0 X 10(12)/L) and decreasing alpha- globin gene number. These data suggest that falling HbF levels among SS individuals with lessened numbers of alpha-globin genes reflect prolonged survival of non-F cells and are not due to intrinsic differences in F cell production or in the amount of HbF per F cell. The improved survival of non-F cells in SS alpha-thalassemia is presumed to be due to the lower MCHC observed in such individuals.
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Dover, GJ, VT Chang, SH Boyer, GR Serjeant, S. Antonarakis i DR Higgs. "The cellular basis for different fetal hemoglobin levels among sickle cell individuals with two, three, and four alpha-globin genes". Blood 69, nr 1 (1.01.1987): 341–44. http://dx.doi.org/10.1182/blood.v69.1.341.bloodjournal691341.
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Fetal hemoglobin (HbF) levels vary widely among individuals with sickle cell anemia (SS). Previous studies have suggested that HbF levels in SS individuals with alpha-thalassemia (two or three functional alpha- globin genes) are lower than HbF levels in SS individuals with four normal alpha-globin genes. Using immunocytochemical techniques, we studied F cell production as measured by % F reticulocytes, the amount of HbF per F cell, and the preferential survival of F cells versus non- F cells in 51 subjects with four alpha genes, 32 subjects with three alpha genes, and 18 subjects with two alpha genes. Comparison between alpha-globin gene groups was performed for the total sample as well as for a subset of 82 individuals who had replicate samples and a further subset of 39 age-matched individuals. %HbF levels were 6.8, 4.9, and 4.5 percent for the total four-, three-, and two-alpha-globin-gene groups, respectively. The percentage of F reticulocytes, percentage HbF per F cell, and the enrichment ratio (% F cell/% F reticulocytes) did not change significantly with alpha-globin gene number. Moreover, no correlation existed between alpha-globin gene number and the absolute number of F cells in any group studied. However, there was a strong inverse correlation (r = -0.407, P = .0001) between non-F cell levels (1.7 +/- 2, 2.2 +/- 5, 3.0 +/- 1.0 X 10(12)/L) and decreasing alpha- globin gene number. These data suggest that falling HbF levels among SS individuals with lessened numbers of alpha-globin genes reflect prolonged survival of non-F cells and are not due to intrinsic differences in F cell production or in the amount of HbF per F cell. The improved survival of non-F cells in SS alpha-thalassemia is presumed to be due to the lower MCHC observed in such individuals.
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Maier-Redelsperger, M., CT Noguchi, M. de Montalembert, GP Rodgers, AN Schechter, A. Gourbil, D. Blanchard, JP Jais, R. Ducrocq i JY Peltier. "Variation in fetal hemoglobin parameters and predicted hemoglobin S polymerization in sickle cell children in the first two years of life: Parisian Prospective Study on Sickle Cell Disease". Blood 84, nr 9 (1.11.1994): 3182–88. http://dx.doi.org/10.1182/blood.v84.9.3182.3182.
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Abstract Intracellular hemoglobin S (HbS) polymerization is most likely to be the primary determinant of the clinical and biologic manifestations of sickle cell disease (SCD). Fetal hemoglobin (HbF) does not enter the HbS polymer and its intracellular expression in sickle erythrocytes inhibits polymerization. HbF levels, high at birth but decreasing thereafter, protect the newborn from the clinical manifestations of this hemoglobinopathy. We have measured the sequential changes in HbF, F reticulocytes, and F cells in the first 2 years of life in 25 children with SCD and compared the results with those obtained in 30 normal children (AA). We have also calculated HbF per F cell (F/F cell), the preferential survival of F cells versus non-F cells, as measured by the ratio F cells versus F reticulocytes (FC/FR) and polymer tendency at 40% and 70% oxygen saturation. HbF levels decreased from about 80.4% +/- 4.0% at birth to 9.2% +/- 2.9% at 24 months. During this time, we observed a regular decrease of the F reticulocytes and the F cells. The kinetics of the decline of F/F cell was comparable with the decline of HbF, rapid from birth (mean, 27.0 +/- 3.6 pg) to 12 months of age (mean, 8.5 +/- 1.5 pg) and then slower from 12 to 24 months of age (mean, 6.2 +/- 1.0 pg) in the SCD children. In the AA children, the decrease in HbF, due to changes in both numbers of F cells and F/F cell, was more precipitous, reaching steady-state levels by 10 months of age. Calculated values for mean polymer tendency in the F-cell population showed that polymerization should begin to occur at 40% oxygen saturation at about 3 months and increase progressively with age, whereas polymerization at 70% oxygen saturation would not occur until about 24 months. These values correspond to HbF levels of 50.8% +/- 10.8% and 9.2% +/- 2.9%, respectively, and F/F cell levels of 15.6 +/- 4.5 pg and 6.2 +/- 1.0 pg, respectively. In the non--F-cell population, polymerization was expected at birth at both oxygen saturation values. Three individuals had significantly greater predicted polymerization tendency than the remainder of the group because of early decreases in HbF. These individuals in particular, the remainder of the cohort, as well as other recruited newborns, will be studied prospectively to ascertain the relationship among hematologic parameters, which determine polymerization tendency and the various clinical manifestations of SCD.
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Maier-Redelsperger, M., CT Noguchi, M. de Montalembert, GP Rodgers, AN Schechter, A. Gourbil, D. Blanchard, JP Jais, R. Ducrocq i JY Peltier. "Variation in fetal hemoglobin parameters and predicted hemoglobin S polymerization in sickle cell children in the first two years of life: Parisian Prospective Study on Sickle Cell Disease". Blood 84, nr 9 (1.11.1994): 3182–88. http://dx.doi.org/10.1182/blood.v84.9.3182.bloodjournal8493182.
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Intracellular hemoglobin S (HbS) polymerization is most likely to be the primary determinant of the clinical and biologic manifestations of sickle cell disease (SCD). Fetal hemoglobin (HbF) does not enter the HbS polymer and its intracellular expression in sickle erythrocytes inhibits polymerization. HbF levels, high at birth but decreasing thereafter, protect the newborn from the clinical manifestations of this hemoglobinopathy. We have measured the sequential changes in HbF, F reticulocytes, and F cells in the first 2 years of life in 25 children with SCD and compared the results with those obtained in 30 normal children (AA). We have also calculated HbF per F cell (F/F cell), the preferential survival of F cells versus non-F cells, as measured by the ratio F cells versus F reticulocytes (FC/FR) and polymer tendency at 40% and 70% oxygen saturation. HbF levels decreased from about 80.4% +/- 4.0% at birth to 9.2% +/- 2.9% at 24 months. During this time, we observed a regular decrease of the F reticulocytes and the F cells. The kinetics of the decline of F/F cell was comparable with the decline of HbF, rapid from birth (mean, 27.0 +/- 3.6 pg) to 12 months of age (mean, 8.5 +/- 1.5 pg) and then slower from 12 to 24 months of age (mean, 6.2 +/- 1.0 pg) in the SCD children. In the AA children, the decrease in HbF, due to changes in both numbers of F cells and F/F cell, was more precipitous, reaching steady-state levels by 10 months of age. Calculated values for mean polymer tendency in the F-cell population showed that polymerization should begin to occur at 40% oxygen saturation at about 3 months and increase progressively with age, whereas polymerization at 70% oxygen saturation would not occur until about 24 months. These values correspond to HbF levels of 50.8% +/- 10.8% and 9.2% +/- 2.9%, respectively, and F/F cell levels of 15.6 +/- 4.5 pg and 6.2 +/- 1.0 pg, respectively. In the non--F-cell population, polymerization was expected at birth at both oxygen saturation values. Three individuals had significantly greater predicted polymerization tendency than the remainder of the group because of early decreases in HbF. These individuals in particular, the remainder of the cohort, as well as other recruited newborns, will be studied prospectively to ascertain the relationship among hematologic parameters, which determine polymerization tendency and the various clinical manifestations of SCD.
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Veraldi, Kristen L., George K. Arhin, Kathleen Martincic, Ling-Hsiu Chung-Ganster, Jeffrey Wilusz i Christine Milcarek. "hnRNP F Influences Binding of a 64-Kilodalton Subunit of Cleavage Stimulation Factor to mRNA Precursors in Mouse B Cells". Molecular and Cellular Biology 21, nr 4 (15.02.2001): 1228–38. http://dx.doi.org/10.1128/mcb.21.4.1228-1238.2001.
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ABSTRACT Previous studies on the regulation of polyadenylation of the immunoglobulin (Ig) heavy-chain pre-mRNA argued fortrans-acting modifiers of the cleavage-polyadenylation reaction operating differentially during B-cell developmental stages. Using four complementary approaches, we demonstrate that a change in the level of hnRNP F is an important determinant in the regulated use of alternative polyadenylation sites between memory and plasma stage B cells. First, by Western analyses of cellular proteins, the ratio of hnRNP F to H or H′ was found to be higher in memory B cells than in plasma cells. In memory B cells the activity of CstF-64 binding to pre-mRNA, but not its amount, was reduced. Second, examination of the complexes formed on input pre-mRNA in nuclear extracts revealed large assemblages containing hnRNP H, H′, and F but deficient in CstF-64 in memory B-cell extracts but not in plasma cells. Formation of these large complexes is dependent on the region downstream of the AAUAAA in pre-mRNA, suggesting that CstF-64 and the hnRNPs compete for a similar region. Third, using a recombinant protein we showed that hnRNP F could bind to the region downstream of a poly(A) site, block CstF-64 association with RNA, and inhibit the cleavage reaction. Fourth, overexpression of recombinant hnRNP F in plasma cells resulted in a decrease in the endogenous Ig heavy-chain mRNA secretory form-to-membrane ratio. These results demonstrate that mammalian hnRNP F can act as a negative regulator in the pre-mRNA cleavage reaction and that increased expression of F in memory B cells contributes to the suppression of the Ig heavy-chain secretory poly(A) site.
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Haller, M., M. Rehm, V. Orth, H. Dressel, H. Brechtelsbauer i U. Finsterer. "HYPERVOLEMIC HEMODILUTION ALTERS THE RATIO OF WHOLE-BODY TO LARGE-VESSEL HEMATOCRIT (f-cell)". Anesthesiology 89, Supplement (wrzesień 1998): 259A. http://dx.doi.org/10.1097/00000542-199809060-00051.
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Bohmer, Ralph M., Thomas A. Campbell i Diana W. Bianchi. "Selectively increased growth of fetal hemoglobin-expressing adult erythroid progenitors after brief treatment of early progenitors with transforming growth factor beta". Blood 95, nr 9 (1.05.2000): 2967–74. http://dx.doi.org/10.1182/blood.v95.9.2967.009k21_2967_2974.
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We have studied the effect of transforming growth factor beta (TGFβ) on erythropoiesis in cultures from adult peripheral blood, using flow cytometric enumeration of fetal hemoglobin (HbF)-containing cells. TGFβ caused a dramatic increase in the proportions of cells that accumulated HbF together with adult hemoglobin (HbA) (F+A+ cells). This highly significant (P < .0001) increase in F+ cell proportion was achieved by TGFβ treatment during the first 4 days of culture and was sustained during further culture expansion in the absence of TGFβ. The increase in F+ cell proportions did not depend on the cytokine combination (EPO+SCF+IL3, EPO+SCF, EPO+IL3, SCF+IL3) used during the phase of TGFβ treatment. Increased F+ cell proportions were paralleled by an increased molecular ratio of HbF/ HbF+ HbA, measured by cation exchange high-performance liquid chromatography (HPLC). In addition to the effect on F+ cell proportions, TGFβ caused a dramatic increase in overall cell division potential. By the time cultures reached terminal growth arrest (12-14 days in controls and 18-26 days after TGFβ), the overall numbers of F+ cells produced per initially seeded clonogenic cell was approximately 10 times higher in the TGFβ-treated cultures than in the controls. We propose to investigate whether the TGFβ-induced increase in relative and absolute numbers of nucleated F+ cells, as demonstrated in vitro, can be translated into increased F+ erythrocytes in vivo, allowing therapeutic application for some beta-hemoglobinopathies.
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Todaro, Maria, Valentina Griggio, Candida Vitale, Chiara Salvetti, Chiara Riganti, Mario Boccadoro, Yosef Landesman i Marta Coscia. "Selinexor in Combination with Chemotherapy or Idelalisib Elicits a Synergistic Cytotoxic Effect in Primary CLL Cells, Also Overcoming Intrinsic and Stromal Cells-Mediated Fludarabine Resistance". Blood 128, nr 22 (2.12.2016): 3210. http://dx.doi.org/10.1182/blood.v128.22.3210.3210.
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Abstract BACKGROUND: Despite the therapeutic efficacy of new target drugs in chronic lymphocytic leukemia (CLL), treatment of high-risk patients remains an unmet clinical need. Disease aggressiveness can be ascribed to intrinsic features of the tumor cells such as immunoglobulin heavy chain (IGHV) mutational status and TP53 disruption, which are strong determinant of drug response. Many of the tumor suppressor and growth regulatory proteins with a known pathogenic role in CLL (i.e. p53, NF-kB, Akt, IkB) bind the nuclear export protein XPO1 (Chromosome Region Maintenance 1; CRM1) and are carried through the nuclear pore complex into the cell cytoplasm. Elevated protein levels of XPO1 and specific XPO1 mutations have been reported in various hematologic and solid tumors. In particular, XPO1 is overexpressed and recurrently mutated in CLL cells (Puente XS et al, Nature 2011; Lapalombella R et al, Blood 2012). Selinexor (KPT-330) an oral inhibitor of XPO1, is active as single agent in different hematologic malignancies including acute myeloid leukemia, non-Hodgkin lymphomas, CLL and multiple myeloma. The combination of selinexor and ibrutinib elicits a synergistic cytotoxic effect in primary CLL cells and increases overall survival of a CLL mouse model compared with ibrutinib alone (Hing ZA et al, Blood 2015). AIM: The aim of this study is to evaluate the additive or synergistic in vitro cytotoxic effects of selinexor, used in combination with chemotherapeutic drugs or the new PI3k inhibitor idelalisib against primary CLL cells. Specifically, this study aims at identifying combination regimens that might overcome single agent resistance. METHODS: 15 patients with CLL were included in the study, among these 9 with mutated (M) and 3 patients with unmutated (UM) IGHV; for 3 patients the mutational status was not available at the moment of data analyses. Purified CLL cells were exposed, alone or in presence of the murine stromal cell line M2-10B4, to selinexor (10 nM, 100 nM, 1 uM and 10 uM) in combination with fludarabine (F-ara-A, 10 nM, 100 nM, 1 uM and 10 uM), bendamustine (Ben, 3 mM, 10 mM, 30 mM and 50 mM) or idelalisib (Ide, 10 nM, 100 nM, 1 uM and 10 uM) for 24, 72 and 120 hours. Cell viability was analysed by Annexin-V/propidium Iodide (AnnV/PI) immunostaining and flow cytometry. Samples were considered resistant when the relative viability of F-ara-A treated CLL cells compared to untreated control was >0.5. Combination analysis was performed using Calcusyn software; combinations were considered synergistic when CI was <1. RESULTS: Leukemic cells were cultured in the presence of increasing concentrations of selinexor, used alone or in combination with F-ara-A and Ben, and with the PI3Kδ inhibitor, Ide. After 72 hours of culture, the mean percentage of viable AnnV-/PI- CLL cells significantly decreased by 0,62-ratio following KPT-330 (100 nM), 0,42-ratio following F-ara-A (1 uM) and 0,24-ratio after dual treatment, compared to untreated controls. Combination analysis showed that selinexor and F-ara-A strongly synergize in inducing CLL cells apoptosis with a CI < 1. Similarly, we observed a synergistic interaction between selinexor (100 nM) and Ben (30 mM) that significantly enhanced the cytotoxic effect of the individual drugs (0,62-ratio for selinexor, 0,68-ratio for Ben and 0,41-ratio for selinexor + Ben) with a CI <1, at the same time point. The combination between selinexor (100 nM) and Ide (10 nM) at 72 hours resulted in a weaker, although significant, viability reduction (0,62-ratio for KPT-330, 0,5 for Ide and 0,38 for selinexor + Ide). We observed that IGHV UM CLL cells showed higher fold reduction values in cell viability when exposed to synergistic combinations, compared to IGHV M cells. Selinexor (100 nM) was also effective in impairing the viability of CLL cells that showed intrinsic resistance to F-ara-A (n=6, 0,61-ratio for selinexor, 0,74-ratio for F-ara-A and 0,4-ratio for selinexor + F-ara-A). Lastly, we exposed CLL-stromal cells co-coltures to the identified synergistic combinations and found that selinexor + F-ara-A or selinexor + Ide significantly reduced the viability of leukemic cells, effectively counteracting the protective effect exerted by stromal cells toward drug-induced apoptosis. CONCLUSIONS: Our data demonstrate that the combination of selinexor with chemotherapy or Ide has synergistic cytotoxic effects, also counteracting intrinsic or stromal cells-mediated drug resistance. Disclosures Boccadoro: SANOFI: Honoraria, Research Funding; CELGENE: Honoraria, Research Funding; Abbivie: Honoraria; Novartis: Honoraria, Research Funding; Amgen: Honoraria, Research Funding; Janssen: Honoraria, Research Funding; BMS: Honoraria, Research Funding; Mundipharma: Research Funding. Landesman:Karyopharm Therapeutics Inc: Employment, Other: stockholder. Coscia:ROCHE: Honoraria, Other: Advisory board; Karyopharm: Research Funding; Mundipharma: Honoraria; Janssen: Honoraria; Gilead: Honoraria.
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Moriichi, Kentaro, Mikihiro Fujiya, Yu Kobayashi, Yuki Murakami, Takuya Iwama, Takehito Kunogi, Takahiro Sasaki i in. "Autofluorescence Imaging Reflects the Nuclear Enlargement of Tumor Cells as well as the Cell Proliferation Ability and Aberrant Status of the p53, Ki-67, and p16 Genes in Colon Neoplasms". Molecules 24, nr 6 (20.03.2019): 1106. http://dx.doi.org/10.3390/molecules24061106.
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Background: Autofluorescence imaging (AFI) is useful for diagnosing colon neoplasms, but what affects the AFI intensity remains unclear. This study investigated the association between AFI and the histological characteristics, aberrant methylation status, and aberrant expression in colon neoplasms. Methods: Fifty-three patients with colorectal neoplasms who underwent AFI were enrolled. The AFI intensity (F index) was compared with the pathological findings and gene alterations. The F index was calculated using an image analysis software program. The pathological findings were assessed by the tumor crypt density, cell densities, and N/C ratio. The aberrant methylation of p16, E-cadherin, Apc, Runx3, and hMLH1 genes was determined by a methylation-specific polymerase chain reaction. The aberrant expression of p53 and Ki-67 was evaluated by immunohistochemical staining. Results: An increased N/C ratio, the aberrant expression of p53, Ki-67, and the altered methylation of p16 went together with a lower F index. The other pathological findings and the methylation status showed no association with the F index. Conclusions: AFI reflects the nuclear enlargement of tumor cells, the cell proliferation ability, and the altered status of cell proliferation-related genes, indicating that AFI is a useful and practical method for predicting the dysplastic grade of tumor cells and cell proliferation.
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Kshivets, O. "Lung cancer prediction: Phase transitions and cell ratio factors". Journal of Clinical Oncology 27, nr 15_suppl (20.05.2009): e22170-e22170. http://dx.doi.org/10.1200/jco.2009.27.15_suppl.e22170.
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e22170 Background: Search of precise prognostic factors for non-small lung cancer (LC) patients (LCP) was realized. Methods: In trial (1985–2008) the data of consecutive 490 LCP after complete resections R0 (age=56.7±8 years; m=439, f=51; tumor diameter: D=4.5±2.1 cm; pneumonectomies=206, lobectomies=284, combined procedures with resection of pericardium, atrium, aorta, VCS, carina, diaphragm, ribs=130; squamous=308, adenocarcinoma=147, large cell=35; T1=143, T2=217, T3=107, T4=23; N0=282, N1=115, N2=93; G1=114, G2=140, G3=236; early LC: LC till 2 cm in D with N0=58, invasive LC=432) was reviewed. Variables selected for 5-year survival (5YS) study were input levels of blood cell subpopulations, TNMG, D. Neural networks computing, Cox regression, clustering, structural equation modeling, Monte Carlo and bootstrap simulation were used to determine any significant regularity. Results: For total of 490 LCP overall life span (LS) was 1824±1304 days (median=1879) and real 5YS reached 62%, 10 years - 50.3%, 20 years - 45.3%. 304 LCP (LS=2597.3±1037 days) lived more than 5 years without LC progressing. 186 LCP (LS=559.8±383.1 days) died because of LC during first 5 years after surgery. 5YS of early LCP was significantly superior (100%) compared with invasive LCP (56.9%) (P=0.000 by log-rank test). 5YS of LCP with N0 was significantly better (78.4%) compared with LCP with N1–2 (39.9%) (P=0.000). Cox modeling displayed that 5YS significantly depended on: phase transition (PT) in terms of synergetics “early-invasive LC”, PT N0-N12, histology, G1–3, cell ratio factors: ratio between the total populations of leucocytes, lymphocytes, neutrophils and LC cells (P=0.000–0.044). Neural networks computing, genetic algorithm selection and bootstrap simulation revealed relationships between 5YS and PT “early-invasive LC”, (rank=1), PT N0-N12 (2), histology (3), G1–3 (4), T1–4 (5), ratio of lymphocytes/LC cells (6), healthy cells/LC cells (7), erythrocytes/LC cells (8), thrombocytes/LC cells (9), eosinophils/LC cells (10), neutrophils/LC cells (11). Correct prediction of 5YS was 100% by neural networks computing (error=0.000; urea under ROC curve=1.0). Conclusions: 5YS of LCP after radical procedures depended on: 1) PT “early-invasive LC”; 2) PT N0-N12; 3) cell ratio factors; 4) LC characteristics. No significant financial relationships to disclose.
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El-Hazmi, Mohsen A. F., Hassan M. Bahakim, Arjumand S. Warsy, Abdulkarim Al-Momen, Abdullah Al-Wazzan, Ibrahim Al-Fawwaz, Sameer Huraib i Mohammad Harakati. "Does G?/A? ratio and Hb F level influence the severity of sickle cell anaemia". Molecular and Cellular Biochemistry 124, nr 1 (1993): 17–22. http://dx.doi.org/10.1007/bf01096377.
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Vande Walle, Johan, Raymond Donckerwolcke, Peter Boer, Hans W. van Isselt, Hein A. Koomans i Jaap A. Joles. "Blood volume, colloid osmotic pressure and F-cell ratio in children with the nephrotic syndrome". Kidney International 49, nr 5 (maj 1996): 1471–77. http://dx.doi.org/10.1038/ki.1996.207.
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Padmanabhan, Krishnanand, Hanna Grobe, Jonathan Cohen, Arad Soffer, Adnan Mahly, Orit Adir, Ronen Zaidel-Bar i Chen Luxenburg. "Thymosin β4 is essential for adherens junction stability and epidermal planar cell polarity". Development 147, nr 23 (1.12.2020): dev193425. http://dx.doi.org/10.1242/dev.193425.
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ABSTRACTPlanar cell polarity (PCP) is essential for tissue morphogenesis and homeostasis; however, the mechanisms that orchestrate the cell shape and packing dynamics required to establish PCP are poorly understood. Here, we identified a major role for the globular (G)-actin-binding protein thymosin-β4 (TMSB4X) in PCP establishment and cell adhesion in the developing epidermis. Depletion of Tmsb4x in mouse embryos hindered eyelid closure and hair-follicle angling owing to PCP defects. Tmsb4x depletion did not preclude epidermal cell adhesion in vivo or in vitro; however, it resulted in abnormal structural organization and stability of adherens junction (AJ) due to defects in filamentous (F)-actin and G-actin distribution. In cultured keratinocytes, TMSB4X depletion increased the perijunctional G/F-actin ratio and decreased G-actin incorporation into junctional actin networks, but it did not change the overall actin expression level or cellular F-actin content. A pharmacological treatment that increased the G/F-actin ratio and decreased actin polymerization mimicked the effects of Tmsb4x depletion on both AJs and PCP. Our results provide insights into the regulation of the actin pool and its involvement in AJ function and PCP establishment.
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Forte, John G., Bernice Ly, Qinfen Rong, Shoji Ogihara, Marlon Ramilo, Brian Agnew i Xuebiao Yao. "State of actin in gastric parietal cells". American Journal of Physiology-Cell Physiology 274, nr 1 (1.01.1998): C97—C104. http://dx.doi.org/10.1152/ajpcell.1998.274.1.c97.
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Remodeling of the apical membrane-cytoskeleton has been suggested to occur when gastric parietal cells are stimulated to secrete HCl. The present experiments assayed the relative amounts of F-actin and G-actin in gastric glands and parietal cells, as well as the changes in the state of actin on stimulation. Glands and cells were treated with a Nonidet P-40 extraction buffer for separation into detergent-soluble (supernatant) and detergent-insoluble (pellet) pools. Two actin assays were used to quantitate actin: the deoxyribonuclease I binding assay to measure G-actin and F-actin content in the two pools and a simple Western blot assay to quantitate the relative amounts of actin in the pools. Functional secretory responsiveness was assayed by aminopyrine accumulation. About 5% of the total parietal cell protein is actin, with about 90% of the actin present as F-actin. Stimulation of acid secretion resulted in no measurable change in the relative amounts of G-actin and cytoskeletal F-actin. Treatment of gastric glands with cytochalasin D inhibited acid secretion and resulted in a decrease in F-actin and an increase in G-actin. No inhibition of parietal cell secretion was observed when phalloidin was used to stabilize actin filaments. These data are consistent with the hypothesis that microfilamentous actin is essential for membrane recruitment underlying parietal cell secretion. Although the experiments do not eliminate the importance of rapid exchange between G- and F-actin for the secretory process, the parietal cell maintains actin in a highly polymerized state, and no measurable changes in the steady-state ratio of G-actin to F-actin are associated with stimulation to secrete acid.
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Krivosova, M., P. Kusnir, M. Kertys, M. Mestanik, I. Tonhajzerova, I. Hrtanek, I. Ondrejka i J. Mokry. "Blood Cell Counts and Blood Cell Ratios as Non-Specific Major Depressive Disorder Biomarkers". Acta Medica Martiniana 19, nr 1 (1.04.2019): 22–29. http://dx.doi.org/10.2478/acm-2019-0003.
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Abstract Introduction: With an increasing prevalence of major depressive disorder (MDD) in population there is a particular interest in finding a suitable biomarker for diagnosis and prognosis of the disease. Many studies have shown that MDD is linked to a systemic inflammatory process, so blood elements counts and ratios have been suggested to be promising indicators in the management and effectiveness of the disease therapy. The aim of this retrospective study was to compare absolute and relative white blood cells counts and to search for any changes in their ratios before and after the therapy of the patients. Methods: Our study included 36 patients who were admitted to hospital with either a new diagnosis or a recurrent episode of MDD and who were treated by a standard protocol. The peripheral blood samples were collected both at admission and at hospital discharge. Absolute white blood cell count and counts of neutrophils, lymphocytes, monocytes, platelets, as well as mean platelet volume, red blood cell distribution width, neutrophil/lymphocyte ratio, platelet/lymphocyte ratio, and monocyte/lymphocyte ratio before and after hospitalization (14–29 days) were evaluated and compared. The test of normality was performed and, accordingly, single t-test or Mann-Whitney U-test was used for data analysis. Results: There were no significant differences between any blood cell ratios in blood samples before and after stay in hospital and appropriate treatment. Monocyte count was significantly higher in MDD patients after hospital discharge (p=0.007), there was a significantly higher difference in discharged patients suffering from MDD recurrent episode (F.33) compared to newly diagnosed MDD (F.32) patients (p=0.010). In patients treated with venlafaxine (N=23) there was a significant increase in monocyte/lymphocyte ratio observed at the end of hospitalization (p=0.018). Conclusions: The pharmacotherapy and additive treatment of the patients suffering from MDD led only to mild changes in blood cells counts. As our study included only a small number of patients, and blood cell parameters and ratios were compared after a relatively short duration of treatment, further and more detailed research is needed for final conclusions.
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Afzal, Asif, Awatef Abidi, Ad Samee, Rk Razak, Manzoore Soudagar i Ahamed Saleel. "Effect of parameters on thermal and fluid flow behavior of battery thermal management system". Thermal Science, nr 00 (2020): 290. http://dx.doi.org/10.2298/tsci191206290a.
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In modern electric vehicles the thermal stability problems associated with Lithium-ion (Li-ion) battery system is of major concern. Proper battery thermal management systems (BTMS) is required to ensure safety and efficient performance of battery cells. A realistic conjugate heat transfer and fluid flow analysis of Li-ion prismatic battery cell is performed. The flow of air as coolant, is laminar, flowing between the heat generating battery cells. The effect of few important working parameters like volumetric heat generation ( q), conduction-convection parameter (?cc), Reynolds number (Re), Aspect ratio (Ar), and spacing between the cells ( f) is investigated in this work. For the wide range of parameters considered, the temperature variations in battery cell and coolant is carried out. Focusing mainly on effect of Re and f, behavior of local Nusselt number (Nux), local friction coefficient (Cf, x), average Nusselt number (Nuavg), average friction coefficient (Cf, avg), maximum temperature, mean fluid temperature, heat removed from the lateral surface of cell are discussed. Nuavg increased with increase in Re but decreased with increase in f, whereas Cf, avg decreased with increase in Re and f. It is also found that their exists an upper and lower limit on Re and f above and below which the change in Cf, avg and Nuavg is negligible. Maximum temperature is significantly influenced at low Re and for all f. From the lateral surface of battery over which the coolant flows, more than 96% of heat generated in cell is removed.
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Araki, N., T. Hatae, T. Yamada i S. Hirohashi. "Actinin-4 is preferentially involved in circular ruffling and macropinocytosis in mouse macrophages: analysis by fluorescence ratio imaging". Journal of Cell Science 113, nr 18 (15.09.2000): 3329–40. http://dx.doi.org/10.1242/jcs.113.18.3329.
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We have applied fluorescence ratio imaging to the analysis of an actin-binding protein concentration relative to F-actin in macrophages, in order to explore the role of a novel (alpha)-actinin isoform, actinin-4, relative to that of the classical isoform, actinin-1. Conventional immunofluorescence images showed that both isoforms were enriched in F-actin-rich regions such as cell surface ruffles. However, ratio images further demonstrated that actinin-4 concentrations relative to F-actin were higher in peripheral inward curved ruffles and dorsal circular ruffles, presumed precursor forms of macropinosomes, than in straight linear ruffles, while actinin-1 concentrations were uniform among the different types of ruffles. Macropinosome pulse-labeling and chase experiments indicated that actinin-4 was also closely associated with newly formed macropinosomes and gradually dissociated with their maturation. Consistent with ratio imaging data, macrophages scrape-loaded with anti-actinin-4 showed a more reduced rate of macropinocytosis than those loaded with anti-actinin-1. Altogether, these results indicate that actinin-4 and actinin-1 contribute differently to F-actin dynamics, that actinin-4 is more preferentially involved in early stages of macropinocytosis than actinin-1. A similar redistribution of actinin-4 was also observed during phagocytosis, suggesting that actinin-4 may play the same role in the two mechanistically analogous types of endocytosis, i.e. macropinocytosis and phagocytosis.
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Köksoy, Gözde T., i F. Dilek Sanin. "Effect of digester F/M ratio on gas production and sludge minimization of ultrasonically treated sludge". Water Science and Technology 62, nr 7 (1.10.2010): 1510–17. http://dx.doi.org/10.2166/wst.2010.447.
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Sludge pretreatment by mechanical, chemical or thermal methods before anaerobic digestion has been applied to increase the digestability of excess sludge. Pretreatment processes rely on their ability to disrupt cell membranes and to release organic materials from the cells into the aqueous phase. Pretreatment by mechanical disintegration has grown rapidly in recent years in parallel with the advances in technology. Ultrasonic sludge disintegration –one of the most commonly used mechanical pretreatment methods- enables the occurrence of cavitation bubbles for the break-up of microorganism cells to extract intracellular materials. The purpose of this study was to conduct disintegration experiments to optimize sonication parameters and to operate subsequent batch anaerobic digesters to examine the effect of food to microorganism ratio (F/M) in sonicated and unsonicated samples. Results showed that high sonication powers and longer treatment times were effective in sludge disintegration in terms of soluble chemical oxygen demand release. Sonicated sludge digested in batch reactors with higher initial F/M ratio caused higher methane generations, higher sludge reductions and had better dewatering characteristics.
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Hatjiharissi, Evdoxia, Daniel Ditzel Santos, Lian Xu, Sigitas Verselis, Michael Modica, Xavier Leleu, Zachary Hunter i in. "Individuals Expressing FcγRIIIA-158 V/V and V/F Show Increased NK Cell Surface Expression of FcgRIIIA (CD16), Rituximab Binding, and Demonstrate Higher Levels of ADCC Activity in Response to Rituximab." Blood 106, nr 11 (16.11.2005): 776. http://dx.doi.org/10.1182/blood.v106.11.776.776.
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Abstract The homozygous expression of phenylalanine (F/F) at codon 158 of FcγRIIIa (CD16) is associated with inferior clinical responses to the anti-CD20 monoclonal antibody rituximab in patients with indolent non-Hodgkin’s lymphoma, in contrast to higher response rates observed for Waldenstrom’s macroglobulinemia patients expressing at least one valine (V/F, V/V) and follicular NHL patients who are homozygous for valine (V/V). We attempted elucidate the potential basic mechanism(s) behind these clinical observations by analyzing all the potential implications of this polymorphism among the 3 polymorphic subgroups at FcγRIIIA-158 (F/F, V/F, and V/V). We therefore used peripheral blood isolated natural killer (NK) cells from a pool of 52 unrelated healthy donors who were genotyped for FcγRIIIA-158. We evaluated allele-specific differences for FcγRIIIa gene expression by quantitative RT-PCR and demonstrated higher transcript levels among V/V (23.2 ng/mL) versus V/F (6.7 ng/mL) and F/F (6.2 ng/mL) (p<0.0001). We then determined protein levels for CD16 in the same subgroup of donors using quantitative flow cytometry. The number of CD16 receptors per NK cell was 105,947, 94,863, and 69,130 for the V/V, V/F and F/F donors, respectively and was significantly higher among donors who expressed at least one valine (V/V and V/F) versus F/F (p=0.033). We next determined rituximab binding affinity to NK cells from V/V, V/F and F/F donors following incubation at concentrations of 10–200 ug/ml, and use of an indirect competitive assay with the anti-CD16 monoclonal antibody 3G8 (Cancer Research 64:4664). Rituximab binding to NK cells was higher among donors expressing at least one valine at all concentrations evaluated, with mean rituximab binding (defined as % of inhibition of mean fluorescence intensity for 3G8) as follow: V/V (72%); V/F (53%); and FF (37%); (p=0.017). Lastly, we assessed rituximab dependent NK cell mediated cytotoxicity (ADCC) using CD20 expressing ARH-77 B-cells and observed higher levels of ADCC killing among V/V (82%) and V/F (80%) versus F/F (23%) donors at an effector: target cell ratio of 20:1. Taken together, these studies suggest that individuals who express at least one valine at FcγRIIIA-158 might have better clinical outcomes to rituximab therapy on the basis of increased FcγRIIIA-158 receptor expression, rituximab binding and ADCC mediated killing of tumor cells.
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Yakoub, Kamal M., Giacomo Lazzarino, Angela M. Amorini, Giuseppe Caruso, Concetta Scazzone, Marcello Ciaccio, Barbara Tavazzi, Giuseppe Lazzarino, Antonio Belli i Valentina Di Pietro. "Fructose-1,6-Bisphosphate Protects Hippocampal Rat Slices from NMDA Excitotoxicity". International Journal of Molecular Sciences 20, nr 9 (7.05.2019): 2239. http://dx.doi.org/10.3390/ijms20092239.
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Effects of fructose 1,6-bisphosphate (F-1,6-P2) towards N-methyl-d-aspartate NMDA excitotoxicity were evaluated in rat organotypic hippocampal brain slice cultures (OHSC) challenged for 3 h with 30 μM NMDA, followed by incubations (24, 48, and 72 h) without (controls) and with F-1,6-P2 (0.5, 1 or 1.5 mM). At each time, cell necrosis was determined by measuring LDH in the medium. Energy metabolism was evaluated by measuring ATP, GTP, ADP, AMP, and ATP catabolites (nucleosides and oxypurines) in deproteinized OHSC extracts. Gene expressions of phosphofructokinase, aldolase, and glyceraldehyde-3-phosphate dehydrogenase were also measured. F-1,6-P2 dose-dependently decreased NMDA excitotoxicity, abolishing cell necrosis at the highest concentration tested (1.5 mM). Additionally, F-1,6-P2 attenuated cell energy imbalance caused by NMDA, ameliorating the mitochondrial phosphorylating capacity (increase in ATP/ADP ratio) Metabolism normalization occurred when using 1.5 mM F-1,6-P2. Remarkable increase in expressions of phosphofructokinase, aldolase and glyceraldehyde-3-phosphate dehydrogenase (up to 25 times over the values of controls) was also observed. Since this phenomenon was recorded even in OHSC treated with F-1,6-P2 with no prior challenge with NMDA, it is highly conceivable that F-1,6-P2 can enter into intact cerebral cells producing significant benefits on energy metabolism. These effects are possibly mediated by changes occurring at the gene level, thus opening new perspectives for F-1,6-P2 application as a useful adjuvant to rescue mitochondrial metabolism of cerebral cells under stressing conditions.
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Di Baldassarre, A., C. Di Rico, T. Bonfini, A. Iacone, M. Marchisio, S. Miscia, E. Alfani, A. R. Migliaccio i G. Migliaccio. "Protein Kinase C (PKC) α Induces Expression of Aγ-Promoter-Controlled Genes in Cellular Models of Hemoglobin Switch." Blood 106, nr 11 (16.11.2005): 3640. http://dx.doi.org/10.1182/blood.v106.11.3640.3640.
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Abstract PKCs are serine-threonine kinases that play an important role in many cellular functions. Almost all the 12 PKC isoforms described so far are expressed throughout erythroid differentiation of human CD34pos cells. Besides the downmodulation of PKCε expression (Bassini et al, Blood93; 1178, 1999), little is known on the role exerted by each PKC on erythroid differentiation. PKCα and PKCδ are two isoforms expressed at comparable levels (as mRNA and protein) throughout differentiation of CD36highCD235alow pro-erythroblasts into orthochromatic erythroblasts. For both proteins, the ratio between phosphorylated and total form remains constant during the differentiation of adult cells. In contrast, the ratio P-PKCα/total PKCα (but not that of P-PKCδ/total PKCδ) increases by 3-fold (with slight donor-to-donor variability) during the differentiation of CD36highCD235alow cells from cord blood. Furthermore, the protein becomes prominently localized in the nucleus of these cells. Since the most stricking difference in the differentiation of adult vs. neonatal erythroblasts is the γ/γ+β globin expression ratio (0.02–0.08 vs. 0.20–0.40 by Taqman, respectively), we hypothesized that the levels of PKCα activity might affect γ-globin expression in erythroblasts. To test this hypothesis, we used two in vitro models of HbF switching. The first model is represented by GM979 cells stably transfected with a dual luciferase (Renilla, R - Firefly, F) reporter driven by either the human γ- or β-globin promoter. The second model is represented by human erythroblasts obtained in HEMA culture. GM979 cells were transiently transfected with expression constructs encoding either the catalytic subunit (sPKCα) or the catalytic inactive (iPKCα) PKCα. The levels of expression of the γ-driven and β-driven luciferase reporters were then measured 24 hrs after transfection. Alternatively, GM979 cells were incubated with concentrations of rottlerin (30 μM) that specifically inhibit PKCα activity. sPKCα increased by 2–3-fold the expression of the luciferase driven by the γ-promoter but did not affect expression from the β promoter. Therefore, the ratio Aγ-F/(Aγ-F+2β-R) was also increased by 2–3 fold. Conversely, rottlerin inhibited (by 50%) both expression of the γ-driven luciferase and the Aγ-F/(Aγ-F+2β-R) ratio. On the other hand, transfection of the double luciferase reporter gene into adult and neonatal CD36highCD235alow cells (30–50% transfection efficiency) resulted in high levels of expression of both reporters, in a ratio consistent with the ontogenic stage of the cells [high Aγ-F/(Aγ-F+2β-R) in neonatal erythroblasts; low Aγ-F/(Aγ-F+2β-R) in adult erythroblasts]. Co-trasfection of sPKCα with this luciferase reporter in CD36highCD235alow cells (both adult and neonatal) increased by 5–6-fold the expression of the luciferase driven from the γ-promoter. Co-trasfection of iPKCα, sPKCδ and iPKCδ had no effect on the expression of the reporters in these cells. In conclusion, using several models of erythroid differentiation, we observed that increased levels of PKCα activity results in increased activity of the γ-promoter, suggesting that element(s) of the haemoglobin switching machinery may represent a specific PKCα substrate in erythroid cells.
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Jadhav, L. D., S. P. Patil, A. P. Jamale i A. U. Chavan. "Solution Combustion Synthesis: Role of Oxidant to Fuel Ratio on Powder Properties". Materials Science Forum 757 (maj 2013): 85–98. http://dx.doi.org/10.4028/www.scientific.net/msf.757.85.
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Solution combustion synthesis technique is one of the novel techniques used to prepare nanoparticles, multi-component ceramic oxides and nanocomposites with properties better than conventionally prepared one and these materials have been used for various applications such as sensors, catalysts, and materials for solid oxide fuel cell (SOFCs). In the present work, the method has been used to prepare nanoparticles of 10 mol% Gd doped ceria (GDC) and Cu and its oxides. The oxidant to fuel (O/F) ratio is found to affect the powder properties and even compositional homogeneity. In glycine-nitrate combustion synthesis of GDC, as revealed by XRD studies, phase pure nanoparticles with crystallite size in the range 9-12nm were obtained for all the O/F ratios. TEM measurements of calcined powder showed hexagonal shaped particles of roughly 20nm size. The exothermicity was increased with the oxidant to fuel ratio resulting in high surface area and soft agglomerates. A slightly lean O/F ratio gives surface area of 73 m2/g and soft agglomerates (D50= 5.34 mm), which eventually results into high sintering density at low temperature. Raman Spectra of GDC showed a sharp and intense peak at 467 cm−1which corresponds to CeO2due to F2gsymmetry of the cubic phase. In combustion synthesis of copper nitrate and citirc acid, the compositional homogenity and phase purity was affected by the oxidant to fuel ratio. The combustion at stoichiometric O/F ratio gives Cu nano particles, lean O/F ratio gives nanoparticles of Cu, CuO and Cu2O and rich ratio gives pure CuO nanoparticles. These nanoparticles have been studied with different characterization techniques like XRD, TG-DTA, SEM, TEM, FT-IR and Raman.
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Koshy, Mabel, Louise Dorn, Linda Bressler, Robert Molokie, Donald Lavelle, Nasrin Talischy, Ronald Hoffman, Wendy van Overveld i Joseph DeSimone. "2-deoxy 5-azacytidine and fetal hemoglobin induction in sickle cell anemia". Blood 96, nr 7 (1.10.2000): 2379–84. http://dx.doi.org/10.1182/blood.v96.7.2379.
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Abstract Augmentation of the fetal hemoglobin (HbF) levels is of therapeutic benefit in patients with sickle cell anemia. Hydroxyurea (HU), by increasing HbF, lowers rates of pain crisis, episodes of acute chest syndrome, and requirements for blood transfusions. For patients with no HbF elevation after HU treatment, augmentation of HbF levels by 5-aza-2′-deoxycytidine (5-aza-CdR, decitabine) could serve as an alternate mode of treatment. Eight adult patients participated in a dose-escalating phase I/II study with 5-aza-CdR at doses ranging from 0.15 to 0.30 mg/kg given 5 days a week for 2 weeks. HbF, F cell, F/F cell, γ-globin synthesis ratio, complete blood count, and chemistry were measured. The average γ-globin synthesis relative to non-α-globin synthesis prior to therapy was 3.19% ± 1.43% and increased to 13.66% ± 4.35% after treatment. HbF increased from 3.55% ± 2.47% to 13.45% ± 3.69%. F cells increased from 21% ± 14.8% to 55% ± 13.5% and HbF/F cell increased from 17% to 24%. In the HU nonresponders HbF levels increased from 2.28% ± 1.61% to 2.6% ± 2.15% on HU, whereas on 5-aza-CdR HbF increased to 12.70% ± 1.81%. Total hemoglobin increased by 1 g/dL in 6 of 8 patients with only minor reversible toxicities, and all patients tolerated the drug. Maximum HbF was attained within 4 weeks of treatment and persisted for 2 weeks before falling below 90% of the maximum. Therefore 5-aza-CdR could be effective in increasing HbF in patients with sickle cell anemia who failed to increase HbF with HU. Demonstration of sustained F levels with additional treatment cycles without toxicity is currently being performed.
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Koshy, Mabel, Louise Dorn, Linda Bressler, Robert Molokie, Donald Lavelle, Nasrin Talischy, Ronald Hoffman, Wendy van Overveld i Joseph DeSimone. "2-deoxy 5-azacytidine and fetal hemoglobin induction in sickle cell anemia". Blood 96, nr 7 (1.10.2000): 2379–84. http://dx.doi.org/10.1182/blood.v96.7.2379.h8002379_2379_2384.
Pełny tekst źródłaStreszczenie:
Augmentation of the fetal hemoglobin (HbF) levels is of therapeutic benefit in patients with sickle cell anemia. Hydroxyurea (HU), by increasing HbF, lowers rates of pain crisis, episodes of acute chest syndrome, and requirements for blood transfusions. For patients with no HbF elevation after HU treatment, augmentation of HbF levels by 5-aza-2′-deoxycytidine (5-aza-CdR, decitabine) could serve as an alternate mode of treatment. Eight adult patients participated in a dose-escalating phase I/II study with 5-aza-CdR at doses ranging from 0.15 to 0.30 mg/kg given 5 days a week for 2 weeks. HbF, F cell, F/F cell, γ-globin synthesis ratio, complete blood count, and chemistry were measured. The average γ-globin synthesis relative to non-α-globin synthesis prior to therapy was 3.19% ± 1.43% and increased to 13.66% ± 4.35% after treatment. HbF increased from 3.55% ± 2.47% to 13.45% ± 3.69%. F cells increased from 21% ± 14.8% to 55% ± 13.5% and HbF/F cell increased from 17% to 24%. In the HU nonresponders HbF levels increased from 2.28% ± 1.61% to 2.6% ± 2.15% on HU, whereas on 5-aza-CdR HbF increased to 12.70% ± 1.81%. Total hemoglobin increased by 1 g/dL in 6 of 8 patients with only minor reversible toxicities, and all patients tolerated the drug. Maximum HbF was attained within 4 weeks of treatment and persisted for 2 weeks before falling below 90% of the maximum. Therefore 5-aza-CdR could be effective in increasing HbF in patients with sickle cell anemia who failed to increase HbF with HU. Demonstration of sustained F levels with additional treatment cycles without toxicity is currently being performed.
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Job, Christy, i Leon Lagnado. "Calcium and Protein Kinase C Regulate the Actin Cytoskeleton in the Synaptic Terminal of Retinal Bipolar Cells". Journal of Cell Biology 143, nr 6 (14.12.1998): 1661–72. http://dx.doi.org/10.1083/jcb.143.6.1661.
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The organization of filamentous actin (F-actin) in the synaptic pedicle of depolarizing bipolar cells from the goldfish retina was studied using fluorescently labeled phalloidin. The amount of F-actin in the synaptic pedicle relative to the cell body increased from a ratio of 1.6 ± 0.1 in the dark to 2.1 ± 0.1 after exposure to light. Light also caused the retraction of spinules and processes elaborated by the synaptic pedicle in the dark. Isolated bipolar cells were used to characterize the factors affecting the actin cytoskeleton. When the electrical effect of light was mimicked by depolarization in 50 mM K+, the actin network in the synaptic pedicle extended up to 2.5 μm from the plasma membrane. Formation of F-actin occurred on the time scale of minutes and required Ca2+ influx through L-type Ca2+ channels. Phorbol esters that activate protein kinase C (PKC) accelerated growth of F-actin. Agents that inhibit PKC hindered F-actin growth in response to Ca2+ influx and accelerated F-actin breakdown on removal of Ca2+. To test whether activity-dependent changes in the organization of F-actin might regulate exocytosis or endocytosis, vesicles were labeled with the fluorescent membrane marker FM1-43. Disruption of F-actin with cytochalasin D did not affect the continuous cycle of exocytosis and endocytosis that was stimulated by maintained depolarization, nor the spatial distribution of recycled vesicles within the synaptic terminal. We suggest that the actions of Ca2+ and PKC on the organization of F-actin regulate the morphology of the synaptic pedicle under varying light conditions.
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Shum, Winnie Waichi, Nicolas Da Silva, Clémence Belleannée, Mary McKee, Dennis Brown i Sylvie Breton. "Regulation of V-ATPase recycling via a RhoA- and ROCKII-dependent pathway in epididymal clear cells". American Journal of Physiology-Cell Physiology 301, nr 1 (lipiec 2011): C31—C43. http://dx.doi.org/10.1152/ajpcell.00198.2010.
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Luminal acidification in the epididymis is critical for sperm maturation and storage. Clear cells express the vacuolar H+-ATPase (V-ATPase) in their apical membrane and are major contributors to proton secretion. We showed that this process is regulated via recycling of V-ATPase-containing vesicles. We now report that RhoA and its effector ROCKII are enriched in rat epididymal clear cells. In addition, cortical F-actin was detected beneath the apical membrane and along the lateral membrane of “resting” clear cells using a pan-actin antibody or phalloidin-TRITC. In vivo luminal perfusion of the cauda epididymal tubule with the ROCK inhibitors Y27632 (10–30 μM) and HA1077 (30 μM) or with the cell-permeable Rho inhibitor Clostridium botulinum C3 transferase (3.75 μg/ml) induced the apical membrane accumulation of V-ATPase and extension of V-ATPase-labeled microvilli in clear cells. However, these newly formed microvilli were devoid of ROCKII. In addition, Y27632 (30 μM) or HA1077 (30 μM) decreased the ratio of F-actin to G-actin detected by Western blot analysis in epididymal epithelial cells, and Y27632 also decreased the ratio of F-actin to G-actin in clear cells isolated by fluorescence activated cell sorting from B1-enhanced green fluorescence protein (EGFP) transgenic mice. These results provide evidence that depolymerization of the cortical actin cytoskeleton via inhibition of RhoA or its effector ROCKII favors the recruitment of V-ATPase from the cytosolic compartment into the apical membrane in clear cells. In addition, our data suggest that the RhoA-ROCKII pathway is not locally involved in the elongation of apical microvilli. We propose that inhibition of RhoA-ROCKII might be part of the intracellular signaling cascade that is triggered upon agonist-induced apical membrane V-ATPase accumulation.
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Liu, M., Z. Huang, Y. Huang, Z. Huang, Q. Huang i T. W. LI. "AB0845 THE COMBINATION OF PLASMA FIBRINOGEN CONCENTRATION AND NEUTROPHIL-LYMPHOCYTE RATIO (F-NLR) AS A NOVEL INFLAMMATORY MARKER OF RHEUMATOID ARTHRITIS". Annals of the Rheumatic Diseases 80, Suppl 1 (19.05.2021): 1446.2–1446. http://dx.doi.org/10.1136/annrheumdis-2021-eular.3929.
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Background:The combined index of fibrinogen and neutrophil-lymphocyte ratio (F-NLR) has recently been reported as a new predictive factor in patients with cancer. However, the fibrinogen and NLR have not been simultaneously evaluated in rheumatoid arthritis (RA).Objectives:This study aimed to explore the clinical value of F-NLR in RA and its relationship with disease activity.Methods:This retrospective study collected 143 RA patients and 82 age- and gender-matched healthy controls. Neutrophil, lymphocyte, monocyte, platelet, fibrinogen, NLR, monocyte to lymphocyte ratio (MLR), platelet to lymphocyte ratio (PLR), C-reactive protein (CRP), erythrocyte sedimentation rate (ESR), Disease Activity Score of 28 joints-ESR (DAS28-ESR) and other laboratory parameters were recorded. Receiver operating characteristic (ROC) curves were used to defined the optimization cut-off values of fibrinogen and NLR, which were 3.9g/L and 2.42. The F-NLR score was 2 for patients with high fibrinogen (> 3.9g/L) and elevated NLR (> 2.42), while those with one or neither were indexed as 1 or 0. The correlations between F-NLR as well as other inflammatory indexes and DAS28-ESR were measured.Results:The F-NLR score was higher in RA patients than that in healthy individuals (P < 0.05). The proportion of higher F-NLR score increased significantly along with the disease activity (P < 0.05). According to the ROC curve which was conducted to discriminate RA patients from healthy subjects, the area under curve (AUC) of F-NLR (0.803, 95% CI: 0.744 - 0.861) was higher than that of fibrinogen (0.735, 95% CI: 0.670 - 0.801), NLR (0.724, 95% CI: 0.655 - 0.794), MLR (0.687, 95% CI: 0.615 - 0.759) and PLR (0.732, 95% CI: 0.664 - 0.800). Furthermore, F-NLR was more strongly associated with DAS28-ESR (r = 0.572, P < 0.001) when compared with fibrinogen (r = 0.518, P < 0.001), NLR (r = 0.365, P < 0.001), MLR (r = 0.140, P = 0.096), PLR (r = 0.239, P = 0.004), CRP (r = 0.539, P < 0.001) and ESR (r = 0.487, P < 0.001).Conclusion:The results demonstrated that the F-NLR score was elevated in RA patients. The F-NLR score may be a potential marker to monitor the disease activity of RA patients.References:[1]Wang H, Zhao J, Zhang M, Han L, Wang M, Xingde L. The combination of plasma fibrinogen and neutrophil lymphocyte ratio (F-NLR) is a predictive factor in patients with resectable non small cell lung cancer. J Cell Physiol. 2018 May; 233(5):4216-4224.Disclosure of Interests:None declared
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Luo, Zhengwei, Jie Zuo, Hui Jiang, Wenhua Geng, Yongzhang Zhou, Zhouyang Lian i Wuji Wei. "Inhibition Effect of Fluoride Ion on Corrosion of 304 Stainless Steel in Occluded Cell Corrosion Stage in the Presence of Chloride Ion". Metals 11, nr 2 (19.02.2021): 350. http://dx.doi.org/10.3390/met11020350.
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The role of F− in the corrosion of stainless steel at the stage of occluded cell corrosion in a mixture of chloride, fluoride, and sulfate ions was investigated. A simulated occluded corrosion cell was designed using an elaborate simulated rust layer. Composite electrodes were used to monitor the variation of the concentration of ions, pH, and dissolved oxygen of the occluded solution. The results show that the influence of F− on the corrosion of 304 stainless steel, in the occluded cell corrosion stage, is concentration dependent. When the F−/Cl− ratio is higher than 2, the corrosion can be significantly suppressed. Analyses showed that the corrosion inhibition effect could be attributed to the migration of F− to the occluded cell, which can reduce the migration of Cl−, dampen the decrease in pH, and react with metal ions to form semi-soluble products. Meanwhile, the influence of F− on the corrosion process was also verified using drilled stainless steel specimens, demonstrating the practicality and validity of the simulated occluded cell corrosion model.
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Pagliaro, L., K. Kerr i D. L. Taylor. "Enolase exists in the fluid phase of cytoplasm in 3T3 cells". Journal of Cell Science 94, nr 2 (1.10.1989): 333–42. http://dx.doi.org/10.1242/jcs.94.2.333.
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We have investigated the intracellular distribution and mobility of the glycolytic enzyme enolase, using functional fluorescent analogs labeled with the succinimidyl esters of carboxyfluorescein (F1-enolase) and carboxytetramethylrhodamine (Rh-enolase) In contrast to aldolase, neither native enolase nor labeled enolase gelled filamentous actin (F-actin), as measured by falling-ball viscometry, indicating a lack of interaction between enolase and F-actin. Fluorescence redistribution after photo-bleaching (FRAP) measurements of the diffusion coefficient (D) of F1-enolase in aqueous solutions gave a value of D37,aq = 6.08 × 10(−7) cm2s-1, and no immobile fraction, consistent with a native molecular weight of 90,000. These values were not significantly different with Rh-enolase, or in the presence of F-actin, 2-phosphoglycerate or F-actin-aldolase gels, demonstrating that neither F1-enolase nor Rh-enolase binds to F-actin or aldolase in vitro. FRAP measurements of F1- and Rh-enolase microinjected into living Swiss 3T3 cells revealed spatial differences in the diffusion coefficient, but not the mobile fraction. In the perinuclear cytoplasm, we measured an apparent diffusion coefficient of 1.1 × 10(−7) cm2s-1, compared to 7.1 × 10(−8) cm2s-1 in the peripheral cytoplasm, with approximately 100% mobility of F1- or Rh-enolase in both regions. Imaging of cells co-injected with Rh-enolase and size-fractionated FITC-dextran (FD-90) revealed that Rh-enolase entered the nucleus, while FD-90 was excluded. Ratio imaging showed a relatively high nuclear ratio of Rh-enolase/FD-90, and a uniform cytoplasmic ratio, with no indication of increased concentration of enolase around stress fibers. These data demonstrate that Rh- and F1-enolase do not bind to F-actin in vitro, and are 100% mobile in vivo. Together with our recent finding that a significant fraction of aldolase binds to F-actin in vitro and is immobile in vivo, these data suggest a correlation between actin-binding activity and cytoplasmic mobility of glycolytic enzymes.
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Togashi, Hideaki, Charles W. Emala, Ian P. Hall i Carol A. Hirshman. "Carbachol-induced actin reorganization involves Gi activation of Rho in human airway smooth muscle cells". American Journal of Physiology-Lung Cellular and Molecular Physiology 274, nr 5 (1.05.1998): L803—L809. http://dx.doi.org/10.1152/ajplung.1998.274.5.l803.
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To determine whether M2 muscarinic receptors are linked to the monomeric G protein Rho, we studied the effect of carbachol on actin reorganization (stress fiber formation) in cultured human airway smooth muscle cells that expressed mainly M2 muscarinic receptors by dual- fluorescence labeling of filamentous (F) and monomeric (G) actin. F-actin was labeled with FITC-labeled phalloidin, and G-actin was labeled with Texas Red-labeled DNase I. Carbachol stimulation induced stress fiber formation (increased F-actin staining) in the cells and increased the F- to G-actin ratio 3.6 ± 0.4-fold (mean ± SE; n = 5 experiments). Preincubation with pertussis toxin, Clostridium C3 exoenzyme, or tyrosine kinase inhibitors reduced the carbachol-induced increase in stress fiber formation and significantly decreased the F- to G-actin ratio, whereas a mitogen-activated protein kinase inhibitor, a phosphatidylinositol 3-kinase inhibitor, and a protein kinase C inhibitor were without effect. This study demonstrates that in cultured human airway smooth muscle cells, muscarinic-receptor activation induces stress fiber formation via a pathway involving a pertussis-sensitive G protein, Rho proteins, and tyrosine phosphorylation.
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Speijer, Dave. "Being right on Q: shaping eukaryotic evolution". Biochemical Journal 473, nr 22 (10.11.2016): 4103–27. http://dx.doi.org/10.1042/bcj20160647.
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Reactive oxygen species (ROS) formation by mitochondria is an incompletely understood eukaryotic process. I proposed a kinetic model [BioEssays (2011) 33, 88–94] in which the ratio between electrons entering the respiratory chain via FADH2 or NADH (the F/N ratio) is a crucial determinant of ROS formation. During glucose breakdown, the ratio is low, while during fatty acid breakdown, the ratio is high (the longer the fatty acid, the higher is the ratio), leading to higher ROS levels. Thus, breakdown of (very-long-chain) fatty acids should occur without generating extra FADH2 in mitochondria. This explains peroxisome evolution. A potential ROS increase could also explain the absence of fatty acid oxidation in long-lived cells (neurons) as well as other eukaryotic adaptations, such as dynamic supercomplex formation. Effective combinations of metabolic pathways from the host and the endosymbiont (mitochondrion) allowed larger varieties of substrates (with different F/N ratios) to be oxidized, but high F/N ratios increase ROS formation. This might have led to carnitine shuttles, uncoupling proteins, and multiple antioxidant mechanisms, especially linked to fatty acid oxidation [BioEssays (2014) 36, 634–643]. Recent data regarding peroxisome evolution and their relationships with mitochondria, ROS formation by Complex I during ischaemia/reperfusion injury, and supercomplex formation adjustment to F/N ratios strongly support the model. I will further discuss the model in the light of experimental findings regarding mitochondrial ROS formation.
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Srinivasan, Srilakshmi, Carson Stephens, Emily Wilson, Janaththani Panchadsaram, Kerry DeVoss, Hannu Koistinen, Ulf-Håkan Stenman i in. "Prostate Cancer Risk-Associated Single-Nucleotide Polymorphism Affects Prostate-Specific Antigen Glycosylation and Its Function". Clinical Chemistry 65, nr 1 (1.01.2019): e1-e9. http://dx.doi.org/10.1373/clinchem.2018.295790.
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Abstract BACKGROUND Genetic association studies have reported single-nucleotide polymorphisms (SNPs) at chromosome 19q13.3 to be associated with prostate cancer (PCa) risk. Recently, the rs61752561 SNP (Asp84Asn substitution) in exon 3 of the kallikrein-related peptidase 3 (KLK3) gene encoding prostate-specific antigen (PSA) was reported to be strongly associated with PCa risk (P = 2.3 × 10−8). However, the biological contribution of the rs61752561 SNP to PCa risk has not been elucidated. METHODS Recombinant PSA protein variants were generated to assess the SNP-mediated biochemical changes by stability and substrate activity assays. PC3 cell–PSA overexpression models were established to evaluate the effect of the SNP on PCa pathogenesis. Genotype-specific correlation of the SNP with total PSA (tPSA) concentrations and free/total (F/T) PSA ratio were determined from serum samples. RESULTS Functional analysis showed that the rs61752561 SNP affects PSA stability and structural conformation and creates an extra glycosylation site. This PSA variant had reduced enzymatic activity and the ability to stimulate proliferation and migration of PCa cells. Interestingly, the minor allele is associated with lower tPSA concentrations and high F/T PSA ratio in serum samples, indicating that the amino acid substitution may affect PSA immunoreactivity to the antibodies used in the clinical immunoassays. CONCLUSIONS The rs61752561 SNP appears to have a potential role in PCa pathogenesis by changing the glycosylation, protein stability, and PSA activity and may also affect the clinically measured F/T PSA ratio. Accounting for these effects on tPSA concentration and F/T PSA ratio may help to improve the accuracy of the current PSA test.
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Noegel, Angelika A., Rosemarie Blau-Wasser, Hameeda Sultana, Rolf Müller, Lars Israel, Michael Schleicher, Hitesh Patel i Cornelis J. Weijer. "The Cyclase-associated Protein CAP as Regulator of Cell Polarity and cAMP Signaling in Dictyostelium". Molecular Biology of the Cell 15, nr 2 (luty 2004): 934–45. http://dx.doi.org/10.1091/mbc.e03-05-0269.
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Cyclase-associated protein (CAP) is an evolutionarily conserved regulator of the G-actin/F-actin ratio and, in yeast, is involved in regulating the adenylyl cyclase activity. We show that cell polarization, F-actin organization, and phototaxis are altered in a Dictyostelium CAP knockout mutant. Furthermore, in complementation assays we determined the roles of the individual domains in signaling and regulation of the actin cytoskeleton. We studied in detail the adenylyl cyclase activity and found that the mutant cells have normal levels of the aggregation phase-specific adenylyl cyclase and that receptor-mediated activation is intact. However, cAMP relay that is responsible for the generation of propagating cAMP waves that control the chemotactic aggregation of starving Dictyostelium cells was altered, and the cAMP-induced cGMP production was significantly reduced. The data suggest an interaction of CAP with adenylyl cyclase in Dictyostelium and an influence on signaling pathways directly as well as through its function as a regulatory component of the cytoskeleton.
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Lee, Jeong Won, Myung Jin Ban, Jae Hong Park i Sang Mi Lee. "Effect of F-18 Fluorodeoxyglucose Uptake by Bone Marrow on the Prognosis of Head and Neck Squamous Cell Carcinoma". Journal of Clinical Medicine 8, nr 8 (4.08.2019): 1169. http://dx.doi.org/10.3390/jcm8081169.
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The purpose of this study was to assess the relationship between F-18 fluorodeoxyglucose (FDG) uptake in bone marrow (BM) on positron emission tomography/computed tomography (PET/CT) and survival in patients with head and neck squamous cell carcinoma (HNSCC). We retrospectively enrolled 157 HNSCC patients who underwent staging FDG PET/CT and subsequent treatment. On PET/CT, primary tumor metabolic characteristics, mean FDG uptake of BM (BM SUV), and BM-to-liver uptake ratio (BLR) were measured. The prognostic significance of FDG uptake of BM for predicting disease progression-free survival and distant failure-free survival was assessed using a Cox proportional hazards regression model. In univariate analysis for disease progression-free survival, increased BM SUV and BLR were associated with poor survival. In multivariate analysis, BLR (p = 0.044; hazard ratio, 1.96), TNM stage (p = 0.014; hazard ratio, 2.87) and maximum FDG uptake of the primary tumor (p = 0.046; hazard ratio, 2.38) were independently associated with disease progression-free survival. For distant failure-free survival, BLR, TNM stage, tumor size, and metabolic parameters of the primary tumor showed prognostic significance in univariate analysis. However, none of the variables showed significance in multivariate analysis. FDG uptake of BM in HNSCC patients might be a significant predictor for disease progression-free survival. Further studies with large patient population are needed to validate the results.
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Lou, Xiaotong, Qianxue Mou, Bowen Zhao, Jingqiu Huang, Ke Yao, Zhaoxia Luo, Meng Ye i in. "VIP Stabilizes the Cytoskeleton of Schlemm’s Canal Endothelia via Reducing Caspase-3 Mediated ZO-1 Endolysosomal Degradation". Oxidative Medicine and Cellular Longevity 2021 (13.09.2021): 1–23. http://dx.doi.org/10.1155/2021/9397960.
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Objectives. In glaucomatous eyes, the main aqueous humor (AH) outflow pathway is damaged by accumulated oxidative stress arising from the microenvironment, vascular dysregulation, and aging, which results in increased outflow resistance and ocular hypertension. Schlemm’s canal (SC) serves as the final filtration barrier of the main AH outflow pathway. The present study is aimed at investigating the possible regulation of vasoactive intestinal peptide (VIP) on the cytoskeleton by stabilizing ZO-1 in SC. Methods. Model of chronic ocular hypertension (COH) induced by episcleral venous cauterization was treated with topical VIP. The ultrastructure of junctions, ZO-1 levels, and permeability of the SC inner wall to FITC-dextran (70 kDa) were detected in the COH models. The F-actin distribution, F/G-actin ratio, and ZO-1 degradation pathway in human umbilical vein endothelial cells (HUVECs) and HEK 293 cells were investigated. Results. ZO-1 in the outer wall of the SC was less than that in the inner wall. COH elicited junction disruption, ZO-1 reduction, and increased permeability of the SC inner wall to FITC-dextran in rats. ZO-1 plays an essential role in maintaining the F/G-actin ratio and F-actin distribution. VIP treatment attenuated the downregulation of ZO-1 associated with COH or H2O2-induced oxidative damage. In H2O2-stimulated HUVECs, the caspase-3 inhibitor prevents ZO-1 disruption. Caspase-3 activation promoted endolysosomal degradation of ZO-1. Furthermore, a decrease in caspase-3 activation and cytoskeleton redistribution was demonstrated in VIP + H2O2-treated cells. The knockdown of ZO-1 or the overexpression of caspase-3 blocked the effect of VIP on the cytoskeleton. Conclusion. This study provides insights into the role of VIP in stabilizing the interaction between the actin cytoskeleton and cell junctions and may provide a promising targeted strategy for glaucoma treatment.
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Nowak, Dorota, Agnieszka Krawczenko, Danuta Duś i Maria Malicka-Błaszkiewicz. "Actin in human colon adenocarcinoma cells with different metastatic potential." Acta Biochimica Polonica 49, nr 4 (31.12.2002): 823–28. http://dx.doi.org/10.18388/abp.2002_3742.
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Four human colon adenocarcinoma cell line variants with different metastatic potential were used to examine whether a correlation exists between actin level, state of actin polymerization and invasiveness of tumour cells. Monomeric (G), total (T) and filamentous (F) actin were determined in the cytosolic fraction of these cells. A statistically significant decrease in G actin level and increase in the state of actin polymerization (measured by F:G actin ratio) were found in the cytosol of three cell variants with higher metastatic potential and invasiveness (EB3, 3LNLN, 5W) compared with the parental cell line (LS180). Our experimental data lead to the conclusion that there is a correlation between the metastatic capacity of human colon adenocarcinoma cells and the state of actin polymerization.
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Mai, Antonello, Silvio Massa, Antonella Di Noia, Katija Jelicic, Elena Alfani, Cristina Di Rico, Angela Di Baldassarre, Anna Rita Migliaccio i Giovanni Migliaccio. "Aroyl-Pyrrolyl-Hydroxy-Amides (APHAs), a Novel Family of Synthetic Histone Deacetylases Inhibitors, Are Potent Inducers of Human g-Globin Gene Expression." Blood 104, nr 11 (16.11.2004): 1216. http://dx.doi.org/10.1182/blood.v104.11.1216.1216.
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Abstract Post-natal pharmacological reactivation of HbF, by restoring the unbalanced α/non-α globin chain production in red cells of patients affected by β-thalassemia or sickle cell anemia, represents a potential cure for these diseases. Many classes of compounds have been identified capable to induce Hb F synthesis in vitro by acting at different levels of the globin gene expression regulatory machinery. One of these classes is represented by inhibitors of a family of enzymes, the histone deacetylases (HDACs), involved in chromatin remodelling and gene transcription regulation. HDACs act in multi-protein complexes that remove acetyl groups from lysine residues on several proteins, including histones and are divided into three distinct structural classes, depending on whether their catalytic activity is zinc (class I/II)- or NAD+ (class III)-dependent. The effects of the HDACs inhibitors identified so far on HbF synthesis is, however, modest and often associated with high toxicity. Therefore, the potential of their clinical use is unclear. We have recently described a new family of synthetic HDACs inhibitors, the Aroyl-pyrrolyl-hydroxy-amides (APHAs), that induce differentiation, growth arrest and/or apoptosis of transformed cell in culture [Mai A et al, J Med Chem2004;47:1098]. In this study, we investigate the capability of 10 different APHA compounds to induce Hb F in two in vitro assays. One assay is based on the ability of APHA compounds to activate either the human Aγ-driven Firefly (Aγ-F) or the β-promoter drives Renilla Luciferase (β-R) reporter in GM979 cells stably transfected with a Dual Luciferase Reporter construct. The second assay is represented by the induction of γ-globin expression (by quantitative RT-PCR) in primary adult erythroblasts obtained in HEMA cultures of mononuclear cells from normal donors. The majority of the compounds tested did not significantly increased the Aγ−F (Aγ−F+β−R) reporter ratio in GM979 cells. However, the compound MC1575 increased by 3-fold (from 0.09 to 0.30) the reporter ratio in GM979 cells at a concentration of 20 μM, with modest effects of the proliferation activity of GM979 cells over the three days of the assay. When MC1575 was added at a concentration of 2–10 μM in cultures of primary adult erythroblasts induced to differentiate in serum-free media for 4 days, it induced a three fold increase of the γ/(γ+β) globin ratio (from 0.04 to 0.12), with no apparent cellular toxicity. Among the HDAC inhibitors tested in this study, MC1575 was not the most potent inhibitor of total enzyme activity. However, it was the compound that most selectively inhibited the activity of the maize homologue of mammalian class IIa HDAC enzymes [Mai et al, J Med Chem2003;46:4826]. These results are consistent with the hypothesis that each class of histone deacetylases might have a specific biological function and indicate that those of class IIa might represent the enzymes most specifically involved in globin gene regulation. We suggest that, by targeting the chemical inhibitors toward the catalytic domain of this class of enzymes, it should be possible to identify more specific, more potent and less toxic compounds for pharmacological treatment of β-thalassemia or sickle cell anemia.
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Wiegand, Marian, Sascha Bossow i Wolfgang J. Neubert. "Sendai virus trailer RNA simultaneously blocks two apoptosis-inducing mechanisms in a cell type-dependent manner". Journal of General Virology 86, nr 8 (1.08.2005): 2305–14. http://dx.doi.org/10.1099/vir.0.81022-0.
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Induction of apoptosis during Sendai virus (SeV) infection has previously been documented to be triggered by initiator caspases (for strain F) or by a contribution of the cellular protein TIAR (T-cell-activated intracellular antigen-related) (for strain Z). Here, evidence was provided that both TIAR and caspases are simultaneously involved in apoptosis induction as a result of infection with SeV strain F. SeV F infection induced death in all tested cell lines, which could only be partially prevented through the pan-caspase inhibitor z-VAD-fmk. However, infection of seven different cell lines with the SeV mutant Fctr48z overexpressing a TIAR-sequestering RNA from the modified leader resulted in a cell type-dependent reduced cytopathic effect (CPE); in an earlier study a similar mutant derived from SeV Z was shown to prevent the induction of any CPE. Finally, blocking of caspases through z-VAD-fmk combined with Fctr48z infection led to complete abrogation of CPE, clearly demonstrating the existence of two separate mechanisms inducing cell death during SeV F infections. Interestingly, a cell type-specific interference between these two mechanisms could be detected during infection with the mutant virus Fctr48z: RNA transcribed from the mutated leader was able to trans-dominantly inhibit caspase-mediated apoptosis. Thus, virus-expressed factors enabling a well-balanced ratio of suppression and triggering of apoptosis seem to be essential for optimal virus replication.
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Holmgren, K., K. E. Magnusson, N. Franki i R. M. Hays. "ADH-induced depolymerization of F-actin in the toad bladder granular cell: a confocal microscope study". American Journal of Physiology-Cell Physiology 262, nr 3 (1.03.1992): C672—C677. http://dx.doi.org/10.1152/ajpcell.1992.262.3.c672.
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Antidiuretic hormone (ADH) induces the fusion of cytoplasmic vesicles containing water channels with the apical membrane of the toad bladder granular cell. Fusion is accompanied by a 30% depolymerization of F-actin. We have used confocal microscopy to determine the region in the cell that undergoes depolymerization. Bladders were mounted in a split chamber, and control halves and halves stimulated by ADH for 15 min were fixed and then stained with rhodamine phalloidin. Vertical sections through the cells were obtained by confocal microscopy, and the fluorescence intensity of the apical and side regions of the cells was determined. To normalize the data, the apex-side intensity was determined for each cell, and these ratios measured for control and ADH-treated halves. In six paired experiments, the ratio for control halves was 3.69 +/- 0.50 and for ADH-treated halves was 2.61 +/- 0.33; the decrease was significant and in good agreement with earlier studies. Thus actin depolymerization takes place in a hormone-sensitive apical pool where vesicle fusion occurs and supports the view that actin depolymerization may be required for fusion.
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Ladygina, L. V., i A. V. Pirkova. "Cultivation of the diatom algae Chaetoceros calcitrans f. pumilus (Paulsen) Takano, 1968 as food for giant oyster larvae Crassostrea gigas (Thunberg)". Marine Biological Journal 4, nr 2 (24.06.2019): 34–40. http://dx.doi.org/10.21072/mbj.2019.04.2.04.
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An impact of modified nutrient media F/2 and Conway on the growth and biomass accumulation of the diatom algae Chaetoceros calcitrans f. pumilus, which is a part of the food for cultivated larvae of the giant oyster Crassostrea gigas in the IMBR RAS nursery, was studied. Maximum values of cell and biomass concentrations were obtained on the modified F/2 nutrient medium (11.22 × 106 cells·ml-1 and 4.93 g·l-1, respectively), and they were much larger than those obtained on Conway medium. Growth parameters of C. calcitrans f. pumilus depended on the ratio of inorganic nitrogen and phosphorus, as well as on the silicon content in nutrient media. The ratio N : P = 12.5 and the silicon concentration of 24 mg·l-1 in the modified F/2 nutrient medium are shown to be approaching the optimal ones for increasing growth rate of diatom algae. It is found that the microalga in concentration 150 × 103 cells·ml-1, cultivated on different nutrient media and included in food composition, has impact on the growth rate of giant oyster larvae. An average daily amount of growth of larvae, whose diet included algae cultivated on modified F/2 nutrient medium, was higher than that of larvae cultivated on Conway medium.
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Kim, Miok, i Minho Shin. "Effect of Educational Program on Knowledge, Attitude, and Willingness of Nursing Students for Hematopoietic Stem-Cell Donation". International Journal of Environmental Research and Public Health 16, nr 19 (1.10.2019): 3696. http://dx.doi.org/10.3390/ijerph16193696.
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This study explored how an educational program on hematopoietic stem-cell donation (HSCD) affects the knowledge, attitude, and willingness for HSCD among nursing students. The subjects were the nursing students at a university in Korea: 43 in the experimental group and 42 in the control group. All subjects took a pre-test, and only the experimental group attended an educational program. Both the groups completed two post-tests. Variables of interest were knowledge, attitude, willingness, and registration ratio for HSCD. The educational program increased knowledge (F = 8.093, p < 0.001) and attitude (F = −6.422, p < 0.001) of the experiment group. After the program, the experimental group showed higher willingness for HSCD (χ2 = 7.609, p = 0.006) and higher registration ratio for HSCD (χ 2= 4.258, p = 0.039) compared to the control group. The educational programs for knowledge and attitude about HSCD will affect the students’ future nursing, and influence clients and their families toward positive perception on HSCD and organ donations.
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Guliński, Piotr, i Anna Kłopotowska. "An attempt to develop a method for determining the typical chemical composition of the milk of Polish Holstein-Friesian cows – a proposal". Roczniki Naukowe Polskiego Towarzystwa Zootechnicznego 15, nr 3 (2.10.2019): 9–21. http://dx.doi.org/10.5604/01.3001.0013.5135.
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The aim of this paper is to develop a method for determining the typical chemical composition of the milk of Polish Holstein-Friesian (PHF) cows. The paper uses data collected from 1329 test-day milking records from 20 herds of PHF dairy cattle in Sokołów County, from 2009 to 2015. The effect of the following factors on the chemical composition of milk was determined: lactation stage (15 one-month stages); age of cows (lactations 1, 2, 3–4, and 5–7); genotype (share of PHF breed: less than 50%, 50–75%, 75–82.5% and more than 82.5%); somatic cell count (SCC) in 1 ml of milk (in thousands: 0–200, 200–400, 400–1000 and more than 1000); feeding level (fat to protein (F/P) ratio): ≤1.0, 1.0–1.4, 1.4–1.7 and >1.7); calving season (autumn/winter, spring/summer) and daily milk yield (milk yield in kg: ≤15, 15–25, 25–35 and >35). Nutrition and udder health status were found to be the main factors influencing the chemical composition of milk. For selected cows with optimally balanced feed rations (F/P ratio in milk from 1.1 to 1.4) and a low somatic cell count (SCC ≤200,000/ml), daily yield was the main factor affecting the chemical composition of the milk. It was also concluded that government and scientific publications on the PHF breed should take into account the impact of the F/P ratio, SCC and yield of milk on its composition.
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Fitzgerald, R. R., i J. D. Walters. "Accumulation of Topical Naproxen by Cultured Oral Epithelium". Journal of Dental Research 86, nr 8 (sierpień 2007): 775–79. http://dx.doi.org/10.1177/154405910708600817.
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Topically administered non-steroidal anti-inflammatory drugs (NSAIDs) inhibit periodontal bone loss, but little is known about the mechanism by which they penetrate oral epithelium. Active transporters could potentially play a role in this process. In this study, we used a cell line derived from oral epithelium to investigate a role for transporters and to characterize conditions that enhance epithelial penetration. Using fluorescence to monitor uptake, we demonstrated that SCC-25 cell monolayers transport naproxen with a Michaelis constant (Km) and maximum velocity (Vmax) of 164 μg/mL and 0.94 ng/min/μg protein, respectively. At steady state, the intra-cellular/extracellular concentration ratio was 3.4. Naproxen accumulation was more efficient at acidic pH than under neutral or alkaline conditions. Small proportions of glycerol, Pluronic F-127, and glucosylceramide enhanced naproxen entry. The individual and combined effects of glycerol and Pluronic F-127 were of lesser magnitude than those obtained with glucosylceramide or at pH 6.3. Thus, SCC-25 cells possess transporters for naproxen.
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Park, Eun K., Maria R. Castrucci, Allen Portner i Yoshihiro Kawaoka. "The M2 Ectodomain Is Important for Its Incorporation into Influenza A Virions". Journal of Virology 72, nr 3 (1.03.1998): 2449–55. http://dx.doi.org/10.1128/jvi.72.3.2449-2455.1998.
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ABSTRACT M2 is an integral protein of influenza A virus that functions as an ion channel. The ratio of M2 to HA in influenza A virions differs from that found on the cell surface, suggesting selective incorporation of M2 and HA into influenza virions. To examine the sequences that are important for M2 incorporation into virions, we used an incorporation assay that involves expressing M2 from a plasmid, transfecting the plasmid into recipient cells, and then infecting those cells with influenza virus. To test the importance of the different regions of the protein (extracellular, transmembrane, and cytoplasmic) in determining M2 incorporation, we created chimeric mutants of M2 and Sendai virus F proteins, exchanging corresponding extracellular, transmembrane, and cytoplasmic domains. Of the six possible chimeric mutants, only three were expressed on the cell surface. Of these three chimeric proteins, only one mutant (with the extracellular domain from M2 and the rest from F) was incorporated into influenza virions. These results suggest that the extracellular domain of M2 is important for its incorporation into virions.