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At least 991 records · Page 55Linked to original sources
PubMed2005

IL-1 alpha stimulates the formation of osteoclast-like cells by increasing M-CSF and PGE2 production and decreasing OPG production by osteoblasts.

Interleukin-1alpha (IL-1alpha) is one of the most potent bone-resorbing factors involved in the bone loss that is associated with inflammation. We examined the effect of the inflammatory mediator IL-1alpha on the expression of macrophage colony-stimulating factor (M-CSF), osteoprotegerin (OPG), and prostaglandin E2 (PGE2) in rat osteoblasts, and the indirect effect of IL-1alpha on the formation of osteoclast-like cells. Osteoblasts were cultured in alpha-minimum essential medium containing 10% fetal bovine serum with or without 100 units/ml of IL-1alpha for up to 14 days. The gene and protein expression of M-CSF and OPG were estimated by determining mRNA levels using the real-time polymerase chain reaction and protein levels using Western blot analysis. PGE2 expression was determined using an enzyme-linked immunosorbent assay. The formation of osteoclast-like cells was estimated using tartrate-resistant acid phosphatase (TRAP) staining of osteoclast precursors in culture with conditioned medium from IL-1alpha-treated osteoblasts and the soluble receptor activator of NF-kappaB ligand (RANKL). M-CSF and PGE2 expression in osteoblasts increased markedly in cells cultured with IL-1alpha, whereas OPG expression decreased. The conditioned medium containing M-CSF and PGE2 produced by IL-1alpha-treated osteoblasts and soluble RANKL increased the TRAP staining of osteoclast precursors. These results suggest that IL-1alpha stimulated the formation of osteoclast-like cells via an increase in M-CSF and PGE2 production, and a decrease in OPG production by osteoblasts.

Animals↗
PubMed2005

Optimal compressive force induces bone formation via increasing bone morphogenetic proteins production and decreasing their antagonists production by Saos-2 cells.

Orthodontic tooth movement induced alveolar bone resorption and formation around the teeth applied mechanical force. Although mechanical force can promote bone formation, the molecular mechanism that underlies this phenomenon is not fully understood. The purposes of this study were to determine how mechanical stress affects the osteogenic response of human osteoblastic cells (Saos-2), and also to examine the optimal compressive force for osteogenesis in vitro. Saos-2 cells were cultured with or without continuously compressive force (0.5-3.0 g/cm2). The expression of bone morphogenetic proteins (BMPs), their antagonists, and transcription factors which involved in osteogenesis were measured using real-time PCR and/or Western blot analysis. Phosphorylation of Smad1 was determined by Western blot. Loading with 1.0 g/cm2 of compressive force significantly increased the expression of BMPs, Runx2 and osterix. In contrast, the expression of BMP antagonists and AJ18 was decreased with 1.0 g/cm2 of compressive force. Loading with 1.0 g/cm2 of compressive force also induced phosphorylation of Smad1. Noggin inhibited the compressive force-induced phosphorylation of Smad1 markedly, and also partially blocked compressive force-induced Runx2 mRNA expression. Moreover, the conditioned medium from 1.0 g/cm2 of compressive force applied cells apparently increased calcium content in mineralized nodules of Saos-2 culture. This study demonstrates that an optimal compressive force stimulates in vitro mineralization via increasing BMPs production and decreasing their antagonists production.

Bone Morphogenetic Proteins↗
PubMed2002

Reactions of alpha amylases with starch granules in aqueous suspension giving products in solution and in a minimum amount of water giving products inside the granule.

Porcine pancreatic alpha amylase (PPA) and Bacillus amyloliquefaciens alpha amylase (BAA) were allowed to react with starch granules from maize, waxy maize, amylomaize-7, and potato in an aqueous suspension with a starch to water ratio of 1:10 and in a minimum of water with a starch to water ratio of 1:1. Quantitative amounts of the maltodextrin products were determined by TLC and scanning densitometry. The two alpha amylases gave different products that were characteristic of their unique action patterns. The percent conversion differed for the different kinds of starches and for the two kinds of reaction conditions. Maize and waxy maize starches were converted into about twice as much maltodextrins than were amylomaize-7 and potato starches by both enzymes and under both reaction conditions. The aqueous suspension gave much greater conversion into maltodextrins than did the minimum water condition. BAA gave 3-14% greater conversion of the granules into maltodextrins than did PPA, with the exception of potato starch.

Animals↗
PubMed2006

Productive and defective impairments in the neglect syndrome: graphic perseveration, drawing productions and optic prism exposure.

The effects of adaptation to prisms displacing rightwards the field of vision on omission errors, and on perseveration and other graphic productions in a line cancellation task, were assessed in nine right-brain-damaged patients with left unilateral spatial neglect. Prism adaptation improved both neglect, as indexed by omission errors, and perseveration behaviour, up to a delay of 60 min. No correlation was found between omission and perseveration errors in all assessments. The suggestion is made that perseveration and other complex graphic productions made by right brain-damaged-patients with left spatial unilateral neglect is due to a defective monitoring of complex motor behaviour, frequently associated to cerebral damage involving the right frontal lobe. Interpretations of perseveration behaviour in terms of allochiria and directional hypokinesia are considered, and their limits discussed.

Adaptation, Physiological↗
PubMed2000

Analysis of human parathyroid hormone (1-84) products. Separation of a major impurity in synthetic products by ion-pairing reversed-phase high-performance liquid chromatography.

Human parathyroid hormone (1-84) is a naturally occurring polypeptide that acts as the major regulator of calcium ion homeostasis. It can be efficiently produced through both synthetic and biosynthetic routes and, as such, highly selective analytical methods are required for the detection of a wide range of impurities. Herein we report on the development of an ion-pairing reversed-phase HPLC method for the analysis of human parathyroid hormone and the separation of impurities including a major, unidentified impurity detected in synthetic preparations. This impurity could not be resolved using trifluoroacetic acid-based methods generally used for monitoring purity levels in commercial products. Separation conditions consisted of a gradient elution of 0.155 M sodium chloride containing 0.037 M sodium pentanesulfonate, pH 5.6, as mobile phase A and acetonitrile as mobile phase B. Separations were carried out on an octadecylsilyl silica column maintained at 50 degrees C. Both column temperature and pH of mobile phase A significantly affected the separation of the major impurity. The major impurity eluted after the main human parathyroid peak and was detected in the two commercial synthetic products analyzed. Several minor impurities eluting before and after the main peak were also detected. Purity levels measured by the developed HPLC method (method C) were similar to those previously measured by capillary electrophoresis. Analysis of purified recombinant human parathyroid hormone did not show the presence of this impurity. This method offers a significant advantage for the purity assessment of human parathyroid hormone.

Chromatography, High Pressure Liquid↗
PubMed1987

Determination of preservatives in cosmetic products. I. Thin-layer chromatographic procedure for the identification of preservatives in cosmetic products.

A thin-layer chromatographic procedure is presented for the separation and identification of preservatives that are listed in the current EEC Council Directive on cosmetic products or have been permitted in the past. The method consists of an extraction of acidified cosmetics with methanol, separation of the extracts by thin-layer chromatography on aluminium oxide and silica gel-coated plates using one developing solvent, and visualization of the preservatives on the plates using short-wavelength UV light and six detection reagents. The retention behaviour and the detectability of 88 preservatives were investigated, of which 74 were characterized by this method. The preservatives in fourteen commercial cosmetic products were tentatively identified by the procedure described. In general this method will permit the routine detection of preservatives in cosmetics in an approximate concentration of 0.1% (w/w).

Chromatography, Thin Layer↗
PubMed1996

Production of pulmonary vasodilation by tolazoline, independent of nitric oxide production in neonatal lambs.

OBJECTIVE: To determine whether tolazoline reduces pulmonary vascular resistance (PVR) by means of endogenous nitric oxide production. DESIGN: Thirty newborn lambs (2 to 7 days of age) were anesthetized with pentobarbital, and their lungs were ventilated through an endotracheal tube. Intravascular catheters were placed in the left ventricle, descending aorta, right atrium, and pulmonary artery for continuous monitoring of intravascular pressures. Cardiac output was measured with radiolabeled microspheres. Arterial carbon dioxide pressure and pH were maintained in a normal range throughout the experiments. Animals were randomly assigned to the following groups: group 1, lungs ventilated with a hypoxic gas mixture and administered tolazoline; group 2, given N omega-nitro-L-arginine (L-NA) (5 mg/min intravenously for 60 minutes) and tolazoline; group 3, given L-NA with hypoxia and tolazoline. Acetylcholine (0.5 microgram/kg) was injected into the right atrium to assess pulmonary nitric oxide synthase activity before and after the L-NA infusion. Data were analyzed by analysis of variance. RESULTS: L-NA inhibited the acetylcholine-induced reduction in mean pulmonary artery pressure (MPAP) by more than 75%. Hypoxia and L-NA increased both MPAP and PVR. Tolazoline produced immediate reductions in both MPAP and PVR in all three groups (group 1, 27% +/- 3% and 50% +/- 5%; group 2, 34% +/- 5% and 50% +/- 6%; and group 3, 31% +/- 4% and 46% +/- 5%, respectively). CONCLUSIONS: These results suggest that tolazoline produces vasodilation independent of nitric oxide production. Understanding the mechanism by which tolazoline produces pulmonary vasodilation may provide insight into the clinical use of this drug and information regarding other potential endogenous mediators of pulmonary vasomotor tone in the neonate.

Animals↗
PubMed1976

Production of Candida utilis on slop by-product of fermentation industries.

The slop (vinas) liquor, a major by-product a alcohol fermentation industries, has been used as growth medium for production of the torula yeast, Candida utilis. Supplementation of the slop with 0.2% ammonium sulphate and 1% to 5% molasses improved the cell yield significantly. The crude slop gave better results than the diluted or centrifuged liquors. Under optimal conditions, more than 15 grams of yeast were obtained on dry weight basis. The application feasibility of these results is presented.

Alcoholic Beverages↗
PubMed1999

Production and breeding of transgenic cattle using in vitro embryo production technology.

Transgenic technology permits major modifications of phenotype by introducing subtle changes in genotype. For domestic farm species, genetic modification may be used to enhance agricultural production or to generate novel genotypes capable of producing heterologous proteins for biomedical applications. The advent of in vitro embryo production techniques has facilitated the large-scale, commercial use of transgenic technology in cattle. Accordingly, we employed in vitro-produced zygotes and embryos in an effort to generate transgenic cattle. Overall, pronuclei in 36,530 in vitro matured and fertilized zygotes were microinjected with a construct designed to express human alpha-lactalbumin in the mammary gland. Of these, 1,472 developed and were transferred to recipients, including 148 twin transfers. Initial pregnancy rate on Day 30 of gestation was 28% (374/1,324). Subsequent calving rate was 17% (226/1,324). Eighteen calves (8%) were transgenic. In vitro produced embryos were used to facilitate breeding of transgenic bulls. Frequency of transgene transmission varied from 3 to 54% between bulls, indicating varying degrees mosaicism. Embryos produced in vitro by these bulls were biopsied and screened for transgenesis prior to transfer to recipients; so far all (6/6) calves born from screened, transgenic embryos were themselves transgenic.

Animals↗
PubMed1985

Effect of bacterial products on prostaglandin E production by amnion cells.

An in-vitro model was set up to study a possible causal relation between bacterial colonisation of extraplacental membranes and spontaneous preterm labour. When amnion cells in tissue culture were exposed to bacterial products, their prostaglandin E output rose considerably. The degree of response varied with the organism. These findings support the theory that bacterial products may be responsible for the premature onset of labour.

Amnion↗
PubMed2000

From gene to product in yeast: production of fungal cutinase.

In the mid-1970s, information technology and recombinant DNA technology were considered as the breakthrough technologies of the final quarter of the 20th century. Now, about 25 years later, information technology has penetrated deeply into our society and nearly everyone uses this technology. Compared to the formidable success of information technology, the progress in the commercialization of recombinant DNA technology is moderate, even when taking into account that all that is related to the technological application of biological sciences needs extensive safety testing. However, there are signs that the speed of this commercialization will increase in the first decade of the 21st century. Moreover, new breakthroughs in our understanding of the complete genetic make up of eukaryotes will contribute to this increase in speed. An important aspect of the commercialization of this technology is the development of cells as factories for the production of valuable and/or useful molecules. Lower eukaryotes, such as yeasts and molds, are the most promising candidates to become the factories of the future, but at present these factories still contains a lot of process lines that may be superfluous under the well controlled conditions in fermentors. On the other hand, the speed and yield of these cellular production lines can be increased by eliminating the rate-determining steps of these process lines. In this contribution to the European Union symposium from Cell to Factory, some steps in the improvement of S. cerevisiae as cell factories for (heterologous) hydrophobic molecules are presented.

Journal Article↗
PubMed1997

Production of human interleukin-8 expressed in Escherichia coli: from a laboratory scale for in vitro tests via a technical scale for animal studies to a process scale for a GMP-compatible production.

An Escherichia coli K 12 strain has been constructed for efficient expression of recombinant biologically active human IL-8 (Interleukin-8). The development of a fermentation and purification process from the laboratory scale (cells from 15 l fermentation broth) to a production scale (cells from 200 l fermentation broth) is described. Material obtained from the laboratory scale was used for initial in vitro studies and for the development of a biological assay. An upscale purification process starting from 80 l fermentation broth resulted in larger amounts of IL-8 needed for preclinical studies. This process includes a fully automated control of the initial affinity chromatography step. Finally, a production process which differed markedly from the small-scale processes was tailor-made for GMP conformity and economic considerations. It consists of a cell disruption step followed by two crossflow diafiltrations with different molecular weight cut offs and filtration rates, one cation exchange chromatography and a final dialysis step. In order to enhance the overall yield of biologically active IL-8, conditions for a resolubilisation of insoluble IL-8 present in the remaining pellet after cell disruption were worked out.

Bioreactors↗
PubMed1978

Production of protein by fungi from agricultural wastes. I. Standardization of certain factors for maximum protein production.

Several fungi were screened for protein production in a simple medium using either sugarcane bagasse, wheat straw or cowdung as substrate. Out of them Rhizoctonia melongina, Pleurotus ostreatus, and Coprinus aratus proved promising and were taken up for further studies. Optimum conditions of temperature, incubation, and pH for maximum growth and protein production by the three fungi were then ascertained and standardized.

Agaricales↗
PubMed2001

Stabilization/solidification of waste ferrous sulphate from titanium dioxide production by fluidized bed combustion product.

A treatment procedure to allow the disposal of waste ferrous sulphate was developed. This waste contains sulphuric acid and originates from titanium dioxide production. The process is based on simultaneous neutralization and stabilization/solidification (S/S) of the waste by means of fluidized bed combustion product (FBC-P). The prepared stabilized waste specimens have a solid matrix and their batch leachates display practically neutral pH and also satisfactory conductivity values. The leachate composition is notable in its absence of toxic metals and even iron.

Coloring Agents↗
PubMed2004

Penicillium expansum: consistent production of patulin, chaetoglobosins, and other secondary metabolites in culture and their natural occurrence in fruit products.

Penicillium expansum is known for its destructive rot and patulin production in apple juice. According to the literature, P. expansum can, among other compounds, produce citrinin, ochratoxin A, patulin, penitrem A, and rubratoxin B. In this study the qualitative production of metabolites was examined using TLC (260 isolates), HPLC (85 isolates), and MS (22 isolates). The results showed that none of the 260 isolates produced ochratoxin A, penitrem A, or rubratoxin B. However, chaetoglobosin A and communesin B were produced consistently by all 260 isolates. Patulin and roquefortine C were produced by 98% of the isolates. Expansolides A/B and citrinin were detected in 91 and 85% of the isolates, respectively. Chaetoglobosins and communesins were detected in naturally infected juices and potato pulp, whereas neither patulin nor citrinin was found. Because most P. expansum isolates produce patulin, citrinin, chaetoglobosins, communesins, roquefortine C, and expansolides A and B, foods contaminated with this fungus should ideally be examined for chaetoglobosin A as well as patulin.

Chromatography, High Pressure Liquid↗
PubMed2006

Determination of carbohydrates in tobacco products by liquid chromatography-mass spectrometry/mass spectrometry: a comparison with ion chromatography and application to product discrimination.

Carbohydrates in commercial tobacco products were quantified utilizing a liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) technique. The method utilizes negative ion electrospray with multiple reactions monitoring and an internal standard calibration. Snuffs, chewing tobaccos, cigars, and cigarettes were analyzed. Product type differentiation was possible using the carbohydrate levels coupled with pH and moisture contents. The LC-MS/MS method was compared to a method utilizing ion chromatography with pulsed amperometric detection. The LC-MS/MS method provided improved selectivity and specificity, demonstrated better precision, and had a larger dynamic range for glucose, fructose, and sucrose in tobacco extracts.

Carbohydrates↗
PubMed2000

Production of stable isotope enriched antimicrobial peptides in Escherichia coli: an application to the production of a 15N-enriched fragment of lactoferrin.

A method is described for the production of recombinant isotopically enriched peptides in E. coli. Peptides are produced in high yield as fusion proteins with ketosteroid isomerase which form insoluble inclusion bodies. This insoluble form allows easy purification, stabilizes the peptide against degradation and prevents bactericidal activity of the peptide. Cyanogen bromide cleavage released peptide which was conjugated with alkylamines to form lipopeptide. An important advantage of this system is that it allows production of peptides that are toxic to bacteria, which we have demonstrated on a dodecapeptide based on residues 21-31 of human bactericidal protein lactoferrin.

Cloning, Molecular↗
PubMed1998

Regulatable production of insulin from primary-cultured hepatocytes: insulin production is up-regulated by glucagon and cAMP and down-regulated by insulin.

To utilize hepatocytes for insulin-producing surrogate cells, we devised a regulatory secretion system by placing proinsulin DNA under the regulatable promoter for phosphoenolpyruvate carboxykinase (PEPCK). The expression of PEPCK is down-regulated by insulin, and up-regulated by cAMP and glucagon. To express insulin in hepatocytes, we constructed an adenoviral insulin expression system. After infection, the hepatocytes secreted immunoreactive insulin (IRI) at an increasing rate. IRI secretion increased over four-fold upon stimulation with 300 microM cAMP and 500 microM of the cAMP-dependent phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine (IBMX). This increase was also observed with glucagon and IBMX. Production was augmented two-fold by the addition of wortmannin, phosphatidylinositol (PI)-3-kinase inhibitor, suggesting that inhibitory insulin signaling to the PEPCK promoter may be mediated through PI-3-kinase. Addition of exogenous insulin to the culture decreased insulin mRNA expression remarkably on Northern blot. Thus, by using a PEPCK promoter for insulin expression, we were able to up-regulate insulin production from hepatocytes with cAMP and glucagon, and down-regulate with insulin itself.

1-Methyl-3-isobutylxanthine↗