Rate Of Cell Multiplication Research Articles

Sustainable organosolv-lignin coated nanosilver-halloysites reinforced poly (vinyl alcohol) nanocomposites for wound healing application

This study involves the fabrication of innovative poly (vinyl alcohol) nanocomposite films by incorporating silver-embedded clay nanotubes with organosolv-lignin by the solution casting approach. The confirmation of this procedure was achieved through the utilisation of many techniques, including FTIR, PXRD, FE-SEM, and TGA. The aforementioned research have verified the adherence of silver nanoparticles to the surface of raw halloysites. The confirmation of lignin functionalization on these nanotubes has been established. This novel nanofiller was used to make a range of nanocomposite films with varying weight percentages ranging from 0 wt% to 5 wt%. With the increase in the wt% of nanofillers, These nanocomposite films exhibited greater thermal and mechanical stability compared to plain PVA. An investigation was conducted to examine the impact of the films on the cellular behaviour of murine fibroblast (NIH3T3) cell lines. Based on the findings from cell proliferation and scratch testing, it has been determined that these nanocomposite films are not harmful to cells, exhibit a greater rate of cell multiplication (116 ± 1.19), and demonstrate increased migratory capabilities (86.5 ± 0.50). Further investigations of human blood corroborate the evidence that these films are compatible with blood. Nanocomposite films have the potential to serve as wound healers following pre-clinical and clinical testing.

Formulation and In vitro Cytotoxicity Studies of Avastin Nanoparticle Mediated Targeted Nose to Brain Drug Delivery System

Nanoparticles (NPs) occupy a superior place among the recent approaches experimented to deliver drugs into brain by circumventing the BBB such as chemical modifications. The Emulsification PEG-PLGA nanoparticles carrying drug were made using the solvent evaporation method. In summary, PEG-PLGA was dissolved in acetone (50 mL) and PLGA was dissolved in PEG-PLGA (50:50, 7-11 mg/mL). The Chitosan-Avastin conjugates were created in the lab in accordance with the procedures and guidelines outlined in Chitosan (3.25 m mol) was hydrated in 1 N HCl, then the appropriate amount of distilled water was added. After adding NaOH (5N) to the reaction mixture to get the pH down to 5, Avastin (6.48 mmol) was added while the mixture was continuously stirred. In-vitro studies the basic aim is to check the effect of various drugs on various cancer cell lines. For testing purpose a large number of human cancer cell lines have been screened. Cells are cultivated in 96-well culture plates, and the rate of cell multiplication, which determines the rate of cell growth, is indirectly measured by the dye's color intensity, which is directly inversely proportional to the number of cells present. Using doses between 10 and 100 g/mL, the MTT test was performed to examine the cytotoxicity various polymer ratios and formulations on human lung cancer lines-A549. For the determination of the percentage of viability of various samples, the cancer cells without receiving therapy were utilized as the control.

Effect of annealing on the mechanical properties and cytocompatibility of CoCrMo alloy with tantalum coating

Cobalt–chromium–molybdenum (CoCrMo) alloy is frequently utilized to replace artificial joints and dental components. Tantalum (Ta) coating is used to improve the osteogenic property of CoCrMo alloy. Annealing treatment can increase the Ta coating adhesion and further improve wear resistance and biocompatibility. In this study, two kinds of thicknesses of Ta coatings were applied on CoCrMo alloy substrates via magnetron sputtering technology. The average thicknesses of the Ta coatings were 4.3 and 9.5 μm. The Ta coatings were annealed for 5 h at 800 °C, 900 °C, and 1000 °C, respectively. Results indicated that heat treatment improved the adhesion of the coatings. The Ta coatings were composed of stable body-centered cubic phase (α-Ta) and metastable tetragonal phase (β-Ta). The Ta coating with a thickness of 4.3 μm exhibited excellent wear resistance after being annealed at 800 °C, with little variability in the friction curve and a low average coefficient of friction. The adhesive force of the two coated samples increased initially and subsequently dropped as the annealing temperature increased. At 900 °C, the Ta-coated samples exhibited the highest adhesive force; after being annealed, the rate of cell proliferation increased. At 900 °C, samples with Ta coatings exhibited the highest rate of cell multiplication and the best cell compatibility compared with the other samples. Ta coatings' adhesive force, wear resistance, corrosion resistance, and cytocompatibility were all significantly enhanced at various annealing temperatures. Thus, the results of this work are a useful benchmark for the surface modification of metal implants used in medical procedures, and Ta coatings are a practical orthopedic material for the replacement of joints and bones.

CD4+ CD25+ regulatory T-cells role in tumor microenvironment of the squamous cell carcinoma.

Introduction: Squamous cell carcinoma (SCC) is the most common skin cancer with a high rate of death. Different lymphocyte populations play an important role in modulating the immune response in the tumor microenvironment. The increase in the proportion of cluster of differentiation (CD)4+ CD25+ regulatory T-cell (Treg) lymphocytes is associated, in different studies, with the increase of the cell multiplication rate. Aim: To analyze the Treg lymphocyte subpopulations and to correlate the results with the presence of the CD8+ cytotoxic T-cell (Tc) lymphocyte population. Materials and Methods: Sixty primary skin SCC specimens were incubated with anti-CD8 (clone SP57) rabbit monoclonal antibody and anti-CD25 (clone 4C9) mouse monoclonal antibody. Results: The ratio of the intratumoral/peritumoral CD4+ CD25+ forkhead box protein p3 (Foxp3) lymphocytes was 0.46, emphasizing that at tumor margins, where tumor aggressiveness is higher, these lymphocytes subpopulations facilitate tumor progression. The comparative analysis of the tumor microenvironment profile revealed that in the case of intratumoral immune response, the number of Tc-type lymphocytes (CD8+) was 3.34 times higher compared to Treg lymphocytes (p<0001). In the peritumoral area, the number of Tc lymphocytes was 5.05 times higher compared to Treg lymphocytes (p<0001). Conclusions: Treg lymphocytes inhibition may cause the suppression of the antitumoral cell immune response in the tumor environment. We believe that Treg lymphocytes should represent a focus of interest for a new personalized therapy. New studies are needed to better understand the immune response in the tumor microenvironment.

Biocompability evaluation of micro textures coated with zinc oxide on Ti-6Al-4V treated by nanosecond laser

Surface morphology of implants plays an important role in enhancing cell response at the interface. Zinc ions (Zn 2+ ) have a promising potential for osseointegration by stimulating osteoblast cell multiplication and differentiation. In this study, regular micro grooves coated with zinc oxide (ZnO) were successfully manufactured on Ti-6Al-4V surface by two-step nanosecond laser treatment to investigate the synergetic effects of micro/nano structures and Zn 2+ on the cell adhesion, proliferation and differentiation. Surface characteristics including surface morphology, roughness, wettability and phase composition were characterized. Results showed that the roughness and hydrophilicity of laser-processing surface were significantly improved compared with those of the acid etching surface. Moreover, XRD results indicated that the ZnO diffraction peak appeared on the surface. The biocompatibility of different samples was investigated by cell experiment in vitro. The spreading morphology of cells along grooves, cell multiplication rate, and activity of alkaline phosphatase and osteocalcin indicated that the micro grooves coated with ZnO on Ti-6Al-4V surface had great potential to promote the adhesion, multiplication and differentiation of MC3T3 cells. This study provides a new and effective method for surface modification of Ti-6Al-4V implants. • A micro textured surface coated with zinc oxide was fabricated on surface of Ti-6Al-4V by two-step nanosecond laser treatment, the precise control of surface morphology and close combination of bioactive coating are realized. • Micro textured surface coated with zinc oxide displayed increased roughness and enhanced hydrophilicity comparing with acid etching surface. • The cell adhesion, proliferation and differentiation of MC3T3s was improved obviously by the synergistic effects of micro grooves and zinc ions released.

In vitro activity of reconstituted rubisco enzyme from Gloeobacter violaceus

RuBisCO (Ribulose 1,5 bisphosphate carboxylase/oxygenase) by virtue of its dual specificity towards oxygen and carbon dioxide is an important rate-limiting step in photosynthesis and is believed to be the key factor for limited productivity of higher plants and algae. The photoautotrophic growth rate of cyanobacteria is a culmination of several factors including, rates of photosynthetic reactions, stress combating mechanisms and basic biomass generation metabolism in combination with optimal nutrient availability, irradiance, gaseous environment, etc. In case of cyanobacteria, the effect of RuBisCO in affecting the multiplication rate has been observed to show varied response. The current paper presents the RuBisCO activity of an early diverging cyanobacterium, Gloeobacter violaceus PCC 7421 and also compares the growth rates and RuBisCO activity of various cyanobacteria. A spectrophotometric estimation in a coupled enzyme assay system of the heterologous expressed G. violaceus PCC 7421 RuBisCO in E. coli, upon purification, revealed a carboxylation activity of LSu to be 5 nMol of phosphoglycerate min-1 mg-1 of protein, which is in coherence with the organism's slow growth rate. Further, the in vitro complementation of RbcL with RbcS in presence of RbcX of G. violaceus facilitated partial reconstitution of the protein and was hence found to cause a four-fold enhancement in its specific activity. The unique characteristics of the primitive cyanobacteria, such as, absence of thylakoids, lack of several photosystem constituting genes, slow carboxylation rate, pose limitation for its rapid multiplication. The RuBisCO carboxylation rate is observed as not the sole but an important parameter for obtaining optimal cell multiplication rates in photo-autotrophically multiplying cyanobacteria.

Temporal Evolution of the Gravitaxis of Euglena gracilis from a Single Cell.

Gravitaxis is one of the most important issues in the growth of microalgae in the water column; it determines how easily cells receive sunlight with a comfortable intensity that is below the damaging threshold. We quantitatively investigated and analyzed the gravitaxis and cell multiplication of Euglena gracilis using vertically placed microchambers containing a single cell. A temporal change in gravitaxis and cell multiplication was observed after transferring the cells to fresh culture medium for 9 days. We performed 29 individual experiments with 2.5 mm × 2.5 mm × 0.1 mm square microchambers and found that the cells showed positive, negative, and moderate gravitaxis in 8, 7, and 14 cases, respectively, after transferring to fresh culture medium. A common trend was observed for the temporal change in gravitaxis for the eight initially positive gravitaxis cases. The cells with initially positive gravitaxis showed a higher rate of cell multiplication than those with initially negative gravitaxis. We also discussed the gravitaxis mechanism of E. gracilis from the observed trend of gravitaxis change and swimming traces. In addition, bioconvection in a larger and thicker chamber was investigated at a millimeter scale and visualized.

Increasing the fermentation efficiency of Lactobacillus paracasei ssp. paracasei MIUG BL6 in a rye flour sourdough

Abstract Background The present study reports the optimization of the biotechnological conditions in order to improve the lactic acid fermentation process, in a rye flour medium by using Lactobacillus paracasei ssp. paracasei strain, coded MIUG BL6, as a starter culture. Materials and methods A sequential statistical methodology, comprised of Plackett-Burman experimental design, central composite design and response surface methodology, was applied to enhance the lactic acid fermentation productivity. Results and discussion Among the tested parameters, the milk powder and KH2PO4 concentration and also the time of fermentation were identified as the most significant variables that influence the fermentation process. The optimum levels of these significant parameters were determined as follows: 4.21% milk powder, 0.30% KH2PO4 and 48 h of fermentation that induced an increase of 1.6 fold of the acidity, compared to the fermentation conducted under the non-optimized conditions. Also, under these optimized fermentation conditions, a good rate of cell multiplication of 10.19 log N/N0 was achieved.

Role of MIF/CD74 signaling pathway in the development of pleural mesothelioma.

Macrophage migration inhibitory factor (MIF) is a pro-inflammatory cytokine implicated in acute and chronic inflammatory diseases. MIF is overexpressed in various tumors. It displays a number of functions that provide a direct link between the process of inflammation and tumor growth. Our group recently identified the MIF-receptor CD74 as an independent prognostic factor for overall survival in patients with malignant pleural mesothelioma.In the present study, we compared the levels of expression of MIF and CD74 in different human mesothelioma cell lines and investigated their physiopathological functions in vitro and in vivo.Human mesothelioma cells expressed more CD74 and secreted less MIF than non tumoral MeT5A cells, suggesting a higher sensitivity to MIF. In mesothelioma cells, high MIF levels were associated with a high multiplication rate of cells. In vitro, reduction of MIF or CD74 levels in both mesothelioma cell lines showed that the MIF/CD74 signaling pathway promoted tumor cell proliferation and protected MPM cells from apoptosis. Finally, mesothelioma cell lines expressing high CD74 levels had a low tumorigenic potential after xenogeneic implantation in athymic nude mice.All these data highlight the complexity of the MIF/CD74 signaling pathway in the development of mesothelioma.

Cytotoxicity investigation of a new hydroxyapatite scaffold with improved structural design

Biodegradable porous scaffolds are found to be very promising bone substitutes, acting as a temporary physical support to guide new tissue regeneration, until the entire scaffold is totally degraded and replaced by the new tissue. The aim of this study was to investigate cytotoxicity of a synthesized calcium hydroxyapatitebased scaffold, named ALBO-OS, with high porosity and optimal topology. The ALBO-OS scaffold was synthesized by the method of polymer foam template. The analysis of pore geometry and scaffold walls’ topography was made by scanning electron microscope (SEM). The biological investigations assumed the examinations of ALBO-OS cytotoxicity to mouse L929 fibroblasts, using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromidefor (MTT) and lactate dehydrogenase (LDH) tests and inverse phase microscopy. The SEM analysis showed high porosity with fair pore distribution and interesting morphology from the biological standpoint. The biological investigations showed that the material is not cytotoxic to L929 cells. Comparison of ALBO-OS with Bio-Oss, as the global gold standard as a bone substitute, showed similar results in MTT test, while LDH test showed significantly higher rate of cell multiplication with ALBO-OS. The scaffold design from the aspect of pore size, distribution, and topology seems to be very convenient for cell adhesion and occupation, which makes it a promising material as a bone substitute. The results of biological assays proved that ALBO-OS is not cytotoxic for L929 fibroblasts. In comparison with Bio-Oss, similar or even better results were obtained.

Dynamische Proliferationsanalysen mittels Durchflusszytometrie

Two key parameters of (eukaryotic) cell culture are cell vitality (survival) and proliferation which both determine the rate of cell multiplication. The proliferation capacity (S-phase fraction) of a given cell population can be easily evaluated by flow cytometry in a static manner. Dynamic proliferation assessment is, however, much more powerful since it reveals profound kinetic information and allows insight into the cell cycle regulation. In particular it is extremely helpful for drug screening.

Measuring bacterial activity and community composition at high hydrostatic pressure using a novel experimental approach: a pilot study.

In this pilot study, we describe a high-pressure incubation system allowing multiple subsampling of a pressurized culture without decompression. The system was tested using one piezophilic (Photobacterium profundum), one piezotolerant (Colwellia maris) bacterial strain and a decompressed sample from the Mediterranean deep sea (3044 m) determining bacterial community composition, protein production (BPP) and cell multiplication rates (BCM) up to 27 MPa. The results showed elevation of BPP at high pressure was by a factor of 1.5 ± 1.4 and 3.9 ± 2.3 for P. profundum and C. maris, respectively, compared to ambient-pressure treatments and by a factor of 6.9 ± 3.8 fold in the field samples. In P. profundum and C. maris, BCM at high pressure was elevated (3.1 ± 1.5 and 2.9 ± 1.7 fold, respectively) compared to the ambient-pressure treatments. After 3 days of incubation at 27 MPa, the natural bacterial deep-sea community was dominated by one phylum of the genus Exiguobacterium, indicating the rapid selection of piezotolerant bacteria. In future studies, our novel incubation system could be part of an isopiestic pressure chain, allowing more accurate measurement of bacterial activity rates which is important both for modeling and for predicting the efficiency of the oceanic carbon pump.

Proteomic analysis of dark response in Arabidopsis cell suspension cultures

Despite intense research on light responses in plants, the consequences of a simple shift from light to darkness are largely unexplored. In this research, the physiological outcome and proteomic changes in Arabidopsis cell suspension cultures after switching from light to total darkness were examined. Deprivation of light led to a visible loss of chlorophyll and failure to develop functional chloroplasts that are present in light-grown cells. This response was accompanied by a significant increase in the cell multiplication rate, most likely due to decreased formation of the damaging reactive oxygen species in the dark. Additionally, there were significant changes in the abundance of 46 protein spots (mostly assigned to chloroplasts, mitochondria and membranes) which were resolved by two-dimensional difference gel electrophoresis and mass spectrometric analysis. All identified chloroplast proteins were down-regulated in response to sustained darkness. In contrast, all differentially expressed proteins associated with cell wall biosynthesis were up-regulated by the dark treatment. Changes in the levels of these proteins were consistent with the observed morphological and physiological changes of the cells. These results reveal a comprehensive picture of the dark response in Arabidopsis cells and provide a useful platform for further characterization of gene function and regulation in plant responses to light.

Glutamylation on α-Tubulin Is Not Essential but Affects the Assembly and Functions of a Subset of Microtubules in Tetrahymena thermophila

Tubulin undergoes glutamylation, a conserved posttranslational modification of poorly understood function. We show here that in the ciliate Tetrahymena, most of the microtubule arrays contain glutamylated tubulin. However, the length of the polyglutamyl side chain is spatially regulated, with the longest side chains present on ciliary and basal body microtubules. We focused our efforts on the function of glutamylation on the alpha-tubulin subunit. By site-directed mutagenesis, we show that all six glutamates of the C-terminal tail domain of alpha-tubulin that provide potential sites for glutamylation are not essential but are needed for normal rates of cell multiplication and cilium-based functions (phagocytosis and cell motility). By comparative phylogeny and biochemical assays, we identify two conserved tubulin tyrosine ligase (TTL) domain proteins, Ttll1p and Ttll9p, as alpha-tubulin-preferring glutamyl ligase enzymes. In an in vitro microtubule glutamylation assay, Ttll1p showed a chain-initiating activity while Ttll9p had primarily a chain-elongating activity. GFP-Ttll1p localized mainly to basal bodies, while GFP-Ttll9p localized to cilia. Disruption of the TTLL1 and TTLL9 genes decreased the rates of cell multiplication and phagocytosis. Cells lacking both genes had fewer cortical microtubules and showed defects in the maturation of basal bodies. We conclude that glutamylation on alpha-tubulin is not essential but is required for efficiency of assembly and function of a subset of microtubule-based organelles. Furthermore, the spatial restriction of modifying enzymes appears to be a major mechanism that drives differential glutamylation at the subcellular level.

Anti‐apoptotic factor humanin is expressed in the testis and prevents cell‐death in leydig cells during the first wave of spermatogenesis

Humanin (HN) is a 24 amino acids peptide with potent neuro-survival properties that protects against damage associated with Alzheimer's disease. In the present report, we have demonstrated by immunohistochemical analysis and Western blotting the pattern of expression of rat humanin (HNr) in the testis of 10- to 60-day-old rats. The Leydig cells of 10- and 40- day-old rats expressed this peptide at high levels; and in the testis of 60-day-old rats the expression of HNr expanded to include Leydig, endothelial, peritubular and germ cells. As monitored by Western blotting, HNr was released into the medium of cultures of Leydig cells isolated from 10-, 40-, and 60-days-old rats. HNr stimulated the incorporation of [(3)H]TdR into DNA of Leydig cells from 10-days-old rats, in a manner that indicated promotion of cell survival rather than an increase in the rate of cell multiplication. This peptide also enhanced steroidogenesis by cultured Leydig cells from 10- to 40-day-old rats both alone and synergistically with IGF-I. The expression of HNr in cultured Leydig cells increased in response to GH and IGF-I. In summary, we demonstrated here that HNr was expressed at all stages of maturation in the rat testis. This peptide promoted the survival of Leydig cells in culture and interacted with IGF-I to stimulate DNA synthesis and steroidogenesis. We propose that HNr is a novel testicular anti-apoptotic factor.

Mixing time in bioreactors under aerated conditions

The major difficulty needing a solution in agitated and aerated bioreactors is closely connected to the optimum hydrodynamics regime in the stirred tank. It is evident that even a simple stirred reactor is geometrically complex and it seems that the mixing time can be one useful criterion for estimation of mixing intensity and therefore the estimation of oxygen profiles and mass transfer coefficients. The mixing time measures are relevant to the length of time for which detectable inhomogeneities last in the extremities of the vessel and can identify the extent of possible concentration gradients in a reactor that should be used as a gradientless unit for bio-kinetic studies. The parameter depends mainly on the mixing system characteristics, the physicochemical properties, the aeration and the rate of cell multiplication. To quantify the influence of some of these factors experiments have been performed using a wide spread laboratory scale stirred dual-impeller bioreactor configuration marketed by “Bioflow”. Experimental data have been obtained for various internal geometries of the reactor under aerated and non-aerated conditions administering real wastewater. A multi-regression analysis method was used to find mathematical correlation relating the influence of the rotation speed and the gas flowrate with the reactor's capacity to mix.

Synergistic anticancer action of reversibly and irreversibly acting ligands of poly (ADP-ribose) polymerase

The synergistic interaction of two ligands (INH2BP and the prodrug INO2BA) of PARP I has been demonstrated for two human leukemia cell lines (855-2 and HL-60), for a human lung cancer cell (A549) and for Eras 20 cancer cells. Synergism was calculated using kinetic combination constants based on cell multiplication rates. Reducing cellular GSH content by BSO strongly augmented synergism, an effect partly explained by the removal of C-NO scavenging (by GSH). However, INH2BP action was augmented by BSO, an effect most probably explained by the sensitization of the cell to apoptosis by GSH removal.

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Wnt-1 inhibits nerve growth factor-induced differentiation of PC12 cells by preventing the induction of some but not all late-response genes

The vertebrate Wnt-1 proto-oncogene is expressed transiently in embryonic brain and functions in the development of the central nervous system and neural crest. The role of Wnt-1 in neural crest development appears to be to increase the number of certain progenitor cells by preventing their premature differentiation. To study the mechanism by which this transient Wnt-1 expression inhibits differentiation we have constructed PC12 pheochromocytoma cells in which Wnt-1 expression levels were controlled by use of a tetracycline-responsive transactivator. Induction of Wnt-1 expression by tetracycline withdrawal was followed by activation of the Wnt-1 signalling pathway as shown by activation of the Lef-1/Tcf transcription factor. Wnt-1 expression by these cells resulted in reversible inhibition of NGF-induced neurite outgrowth, but it did not adversely affect the maintenance of previously formed NGF-induced neurites. Wnt-1 expression also partially blocked the ability of NGF to decrease the rate of cell multiplication. Wnt-1 decreased the NGF-induced expression of the late-response gene SCG10 but not of the immediate early genes, fos, Nur77 and UPAR (urokinase-type plasminogen activator receptor) nor of the late-response genes GAP-43 and collagenase. The Wnt-1 expressing PC12 cells multiplied at a greater rate when they expressed Wnt-1 than they did in the absence of Wnt-1 expression, a result that is consistent with the proposal that Wnt-1 may also act as a mitogen.

Development of an optimal bacterial medium based on the growth inhibition assay with Vibrio fischeri

Chronic toxicity level of chemicals to bacteria can depend on composition and concentration of medium ingredients. This was demonstrated by means of a growth rate inhibition test with the marine bacterium V. fischeri. In a minimal medium (following the validity criterion of achieving a least cell multiplication rate) containing only yeast extract as organic nutrient component, V. fischeri was to Cu 2+ 11,9 times, to Hg 2+ 3 times and to Zn 2+ 2,8 times more sensitive than in the complete defined medium. Obviously yeast extract did not interfere greatly in the complexation mechanisms. The detection limits of the toxicity of Cd 2+ 3,5-dichorophenol and nitrobenzene were not influenced by the substrate.