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PubMedDate not supplied

Temperature reduction in cultures of hGM-CSF-expressing CHO cells: effect on productivity and product quality.

We have demonstrated that temperature reduction from 37 to 33 degrees C in the culture of a CHO cell line producing recombinant human granulocyte macrophage colony stimulating factor (CHO-K1-hGM-CSF) leads to a reduced growth rate, increased cell viability, improved cellular productivity, and decreased cell metabolism. In the present study, CHO-K1-hGM-CSF cells were cultured in a biphasic mode: first, a 37 degrees C growth phase for achieving a high cell number, followed by a production phase where the culture temperature was shifted to 33 degrees C. The maximum cell density was not affected after temperature reduction while cell viability remained above 80% for a further 3.7 days in the culture kept at the lower temperature, when compared to the control culture maintained at 37 degrees C. Furthermore, the total rhGM-CSF production increased 6 times in the culture shifted to 33 degrees C. Because the quality and hence the in vivo efficacy of a recombinant protein might be affected by numerous factors, we have analyzed the N- and O-glycosylation of the protein produced under both cell culture conditions using high-pH anion-exchange chromatography and complementary mass spectrometry techniques. The product quality data obtained from the purified protein preparations indicated that decreasing temperature had no significant effect on the rhGM-CSF glycosylation profiles, including the degree of terminal sialylation. Moreover, both preparations exhibited the same specific in vitro biological activity. These results revealed that the employed strategy had a positive effect on the cell specific productivity of CHO-K1-hGM-CSF cells without affecting product quality, representing a novel procedure for the rhGM-CSF production process.

Animals↗

PubMed2003

Disassembly factories for electrical and electronic products to recover resources in product and material cycles.

Cycle economy as a paradigm for industry in the 21st century depends on the economical and ecological treatment of limited resources. The objective is to achieve more use with fewer resources to increase the use-productivity of these resources. The European Union, aware of the adverse environmental impacts associated with electrical and electronic consumer goods in particular, has passed legislation regulating their appropriate end-of-life treatment. Adaptation processes, including essential disassembly and re-assembly operations, contribute significantly toward the economical fulfillment of these new legal requirements. Typically, the disassembly of used products is characterized by a high rate of manual operations, wide variety of product types, and unknown product properties. To cope with such demands, life cycle units or product accompanying information systems, are being developed and used for acquiring data about a specific product throughout its life cycle to aid in determining the level of product deterioration. Modular disassembly processes and tools have been developed and realized to enable the handling of multiple productvariants. They are being implemented in prototypical hybrid disassembly systemsfor large- and small-size electrical and electronic consumer goods.

Conservation of Energy Resources↗

PubMed1997

The influence of membrane fluidity, TNF receptor binding, cAMP production and GTPase activity on macrophage cytokine production in rats fed a variety of fat diets.

The effects of different dietary fats on peritoneal macrophage plasma membrane fluidity, intracellular cyclic AMP (cAMP) production, GTP hydrolysis and TNF binding and TNF-induced IL1 and IL6 production was investigated. After a four week period, fluidity, as determined by both fluorescence recovery after photobleaching (FRAP) and anisotropy was lowest and highest in animals fed corn and fish oil respectively. After eight weeks feeding, lateral membrane movements were decreased substantially in fish, olive and coconut oil fed dietary groups, whereas an increase in the corn oil fed group was observed, no effect was observed in macrophages from the butter fed group. However, an increase in the packing was observed in macrophages from all dietary groups except in the olive oil fed group. GTPase values for the coconut oil and butter groups were higher than in any other dietary group. After receiving the diet for 8 weeks these differences between the groups were no longer apparent. Exposure of macrophages to TNF had no effect on the rate of GTP hydrolysis. A major enhancement of cAMP production became apparent between weeks 4 and 8 of dietary treatment. After 4 weeks on the diet, values were significantly higher from cells of animals fed corn and olive oils than from animals fed fish oil. After 8 weeks, while there was a general enhancement of production, further differences became apparent. Feeding corn and coconut oils resulted in the highest values and olive oil and chow in the lowest. It is proposed that fats rich in n-3 fatty acids (fish oils) alter membrane fluidity, decrease TNF binding affinity, GTPase activity and cAMP production which appears not to modify cytokine production after short term dietary supplementation. However, after long term feeding it appears that increases in the sensitivity of the TNF receptors plays a major role in modifying cytokine production.

Animals↗

PubMed2002

Effect of temperature on longevity and productivity in Drosophila ananassae: evidence for adaptive plasticity and trade-off between longevity and productivity.

The genetic response of body size to temperature in the laboratory provides an interesting example of phenotypic plasticity. We found that females of Drosophila ananassae reared to adulthood at 18 degrees C showed significant increase in body weight as compared to females reared at 25 degrees C. At a given temperature, early productivity and lifetime productivity were the highest when the rearing and test temperature were the same. The effect of test temperature was highly significant for total productivity and early productivity. The interaction between test temperature and development temperature was also highly significant. Effect of development temperature was not significant. The females reared at 18 degrees C showed greater body weight but their productivity was not significantly higher than smaller females reared at 25 degrees C. Thus, the usually close relationship between size and fecundity is lost when the size change is due to rearing temperature. These findings provide evidence for adaptive plasticity in D. ananassae. We also found a negative correlation (trade-off) between longevity and productivity, the first report of such a trade-off between longevity and productivity in D. ananassae.

Adaptation, Physiological↗

PubMed1992

IL-1 production as a regulator of G-CSF and IL-6 production in CSF-producing cell lines.

We previously demonstrated that colony stimulating factor (CSF)-producing cell lines co-produce interleukin-1 (IL-1) and IL-6 in addition to CSFs. In the present study, we examined the role of IL-1 production in three human tumour cell lines producing granulocyte (G)-CSF, IL-1 and IL-6. Addition of anti-human IL-1 alpha antiserum to the culture caused a 90-62% reduction of G-CSF and a 85-44% reduction of IL-6 production, respectively, as evaluated by enzyme immunoassay in all three cell lines. The decrease of G-CSF and IL-6 production by the anti-IL-1 alpha antiserum was also confirmed at the level of mRNA expression. The anti-IL-1 alpha antiserum did not affect the growth of these cell lines. Excess recombinant IL-1 alpha exogenously added to the culture enhanced G-CSF and IL-6 production in all three cell lines. However, IL-1 alpha had little effect on the growth of these three cell lines. Neither anti-IL-6 nor anti-G-CSF antibodies affected the production of the other cytokines. These results indicate that IL-1 alpha regulates G-CSF and IL-6 production in these tumour cell lines, and suggest that the IL-1 production plays an important role in CSF-producing tumours.

Cell Division↗

PubMed2002

Potential microbiological hazards in the production of refined paper products for food applications.

This study sought to investigate the significance of raw materials (starch-based glues, raw material papers) at different microbiologically critical stages in the manufacturing process of refined paper products. The study examined the occurrence of microorganisms in the process and in end-product samples. Microbiological surveys verified that the production and use of pasteurized starch-based glue was the most important factor threatening the process hygiene and product safety. Subsequently, the production and use of starch-based glue was changed, and a follow-up programme targeting the microbiological quality of glue was developed as part of a hygiene and safety management system. A total of 33 spore-forming bacterial and 15 enterobacterial isolates were ribotyped, and 22 and 10 different ribogroups (ribotypes), respectively, were generated. These isolates from starch-based glue, raw material paper and end products were atypical and, thus, in many cases physiological, chemotaxonomic (FAME) and molecular (partial 16S rDNA) results did not correspond. The most common spore-forming bacteria (55% of the isolates) were Paenibacillus sp. and within this genus several new species were also proposed. The most common enterobacteria (87%) were Enterobacter cloacae and Citrobacter freundii belonging to bacteria in hazard group 2, or species closely related to them. It was demonstrated that the same spore-forming bacteria (ribotypes) were present in both the glue samples and the end products (45% of isolates). All RiboPrint patterns were saved at the VTT identification library for future use.

Adhesives↗

PubMed1998

Differential production of IL-10 by T cells and monocytes of HIV-infected individuals: association of IL-10 production with CD28-mediated immune responsiveness.

Immune unresponsiveness in HIV-1 infection can result from impaired signals delivered by the costimulatory CD28-B7 pathway and the altered production of immunoregulatory cytokines, in particular IL-10, whose production is altered in HIV-1 infection. In this study we investigate IL-10 regulation in T cells and monocytes from HIV+ individuals, and its association with CD28-mediated T cell proliferation. IL-10 production as analysed in T cell- and monocyte-depleted peripheral blood mononuclear cells (PBMC), and by intracellular staining at the single-cell level, reveals a defect in IL-10 production by CD4+ and CD8+ T cells, whereas monocytes constitute the major IL-10-producing cell type. To investigate the impact of IL-10 on immune responsiveness, CD28-mediated proliferative responses in HIV+ individuals were correlated with PHA-induced IL-10 production. CD4+ T cells expressed CD28, yet exhibited markedly reduced CD28-mediated cell proliferation. This CD28-mediated CD4+ T cell proliferation was found to be inversely associated with the levels of PHA-induced IL-10 production and could be restored, at least in part, by anti-IL-10 antibodies. These results suggest that IL-10 production is differentially regulated in T cells and monocytes of HIV+ individuals, and that IL-10 may have a role in inducing immune unresponsiveness by modulating the CD28-B7 pathway.

Adult↗

PubMed2000

Proteolytic activity of cysteine protease in excretory-secretory product of Paragonimus westermani newly excysted metacercariae pivotally regulates IL-8 production of human eosinophils.

This study investigated the effect of the excretory-secretory product (ESP) of Paragonimus westermani newly excysted metacercariae (PwNEM) on IL-8 production of human mature eosinophils. Treatment of eosinophils with lower concentrations (0.3 and 1 microg/ml) of the ESP significantly (P < 0.01) induced IL-8 production, whereas treatment of cells with higher concentrations (3 and 10 microg/ml) did not. This effect of the ESP was concentration-dependent. Interestingly, the amount of IL-8 production released into the culture supernatants was inversely correlated with the rate of eosinophil survival. When eosinophils were cultured with the same concentrations of the ESP for 24 h, the ESP resulted in eosinophil death in a dose-dependent manner. To investigate whether high proteolytic activity of proteases in the ESP could cause little production of IL-8, 10 microg/ml of ESP was pretreated with heat at 100 degrees C for 10 min or 56 degrees C for 30 min to reduce its proteolytic activity. IL-8 production of eosinophils incubated with heat-treated ESP was markedly increased comparable to that of cells treated with the lowest concentration used in this study. These findings suggest that the protease in the ESP of PwNEM pivotally regulates IL-8 production by controlling of eosinophil survival, depending on the amount of ESP released in vivo. Thus, the cysteine protease in the ESP of PwNEM could provide a novel role to control recruitment of inflammatory cells in larval-infected lesions in human paragonimiasis.

Animals↗

PubMed2003

[Quality control of gene therapy products: approach of the French Agency for the Safety of Health Products].

Gene therapy is a new therapeutic strategy which can constitute in some diseases a true alternative or a complement to the "classical treatments". Regarding the innovative features, the complexity and the extreme diversity of the gene therapy products (naked DNA, synthetic vectors, viral vectors, genetically modified cells), these new products presently in clinical trials have to be precisely evaluated and controlled for their medicine quality as well as their biological origin and/or their specific characteristics of genetically modified organisms. The French Agency for the Safety of Health Products engaged an in-depth scientific review concerning the control of this very heterogeneous class of potential therapeutics through the creation of a working group. The objectives of this group were to determine the testings to be performed by a national authority for each type of gene therapy products and to select the appropriate techniques or methods to be developed. Controls considered as essential are listed and include the verification of the identity, the purity, the transfer and expression efficiency as well as the microbiological and viral safety of the products. This implies the development of diverse techniques of molecular biology, cellular biology, physico-chemistry, animal testing, histology and microbiology. Finally, in order to define the basis of testings of these emerging products, the marketing of which should be effective for some of them in the next years, it appears extremely important to harmonize the quality, efficiency and safety criteria, to develop specific references and standards and to create specific guidelines for the control of gene therapy products.

France↗

PubMed1997

Induction of airway mucus production By T helper 2 (Th2) cells: a critical role for interleukin 4 in cell recruitment but not mucus production.

Airway inflammation is believed to stimulate mucus production in asthmatic patients. Increased mucus secretion is an important clinical symptom and contributes to airway obstruction in asthma. Activated CD4 Th1 and Th2 cells have both been identified in airway biopsies of asthmatics but their role in mucus production is not clear. Using CD4 T cells from mice transgenic for the OVA-specific TCR, we studied the role of Th1 and Th2 cells in airway inflammation and mucus production. Airway inflammation induced by Th2 cells was comprised of eosinophils and lymphocytes; features found in asthmatic patients. Additionally, there was a marked increase in mucus production in mice that received Th2 cells and inhaled OVA, but not in mice that received Th1 cells. However, OVA-specific Th2 cells from IL-4-deficient mice were not recruited to the lung and did not induce mucus production. When this defect in homing was overcome by administration of TNF-alpha, IL-4 -/- Th2 cells induced mucus as effectively as IL-4 +/+ Th2 cells. These studies establish a role for Th2 cells in mucus production and dissect the effector functions of IL-4 in these processes. These data suggest that IL-4 is crucial for Th2 cell recruitment to the lung and for induction of inflammation, but has no direct role in mucus production.

Adoptive Transfer↗

PubMed1998

Feeding diets containing high levels of milk products or cellulose decrease urease activity and ammonia production in rat intestine.

Three studies were done to determine the effect of feeding diets containing high levels of a readily fermentable carbohydrate (lactose in milk or yogurt, or pure lactose) or an undigestible, unfermentable diluent (alpha-cellulose) on urease (EC 3.5.1.5) activity and net ammonia production in the rat gastrointestinal (GI) contents. Rats (170-200 g) were fed a control diet or diets containing 55% dried milk or 55% dried yogurt, 25% lactose or 10% alpha-cellulose. Feeding diets containing milk or yogurt decreased urease activity to approximately 11% of the control value in the small intestine (on the basis of grams of collected contents or total contents), and to 50% in the large intestine (only on the basis of grams of collected contents). Feeding the diet containing 25% lactose also decreased urease activity (on the basis of grams of collected contents or total contents) to about 20% of the control value in the small intestine, but not (P > 0.05) in the large intestine. Net ammonia production rate was correlated (r2 = 0.98) with urease activity in the large intestinal contents, and the rate of ammonia production from ureolysis represented about two thirds of the total. Feeding the cellulose diet decreased (P < 0.05) both urease activity and net ammonia production in the large intestine to approximately 30% of the control value. Weights of tissue and contents of the large intestine were much higher (P < 0.01) in rats fed diets containing milk products or lactose than in the control rats, but were not affected by consumption of the cellulose diet. Results of our studies indicate that feeding diets containing high levels of milk products (lactose) or cellulose reduces urease activity and net ammonia production in the rat intestine, and thus may be beneficial for improving animal and human health.

Ammonia↗

PubMed2006

Effect of ascorbate and its oxidation products on H2O2 production in cell-suspension cultures of Picea abies and in the absence of cells.

Dehydroascorbate and traces of ascorbate were present apoplastically in living spruce (Picea abies) twigs. Since the proposed apoplastic ascorbate degradation pathway contains several steps that possibly generate H(2)O(2), the effects of ascorbate and some of its degradation products were tested on apoplastic H(2)O(2) concentrations in a cell culture of P. abies as a model and on non-enzymic H(2)O(2) production in vitro. Ascorbate scavenged H(2)O(2) in the culture medium of lignin-producing Picea cells and in spent and boiled spent medium; in the presence of Cu(2+) or fresh medium, ascorbate led to the non-enzymic generation of H(2)O(2). Preparations of dehydroascorbate (the initial oxidation-product of ascorbate), and diketogulonate (the hydrolysis-product of dehydroascorbate) induced H(2)O(2) accumulation both non-enzymically and enzymically in Picea cell-suspensions. Paper electrophoresis showed that the dehydroascorbate and diketogulonate preparations contained several degradation products; some of these probably contributed to H(2)O(2) production and/or scavenging in these experiments, and would also do so in vivo. These results indicate a complex ability of apoplastic ascorbate, dehydroascorbate, diketogulonate, and further products to modulate H(2)O(2) concentrations, with potential consequences for the control of growth, development and lignification.

2,3-Diketogulonic Acid↗

PubMed2002

Genetics of growth and reproduction in the turkey. 15. Effect of long-term selection for increased egg production on the genetics of growth and egg production traits.

A line (E) of turkeys selected long-term (37 generations) for increased egg production was reciprocally crossed with its randombred control population (RBC1) that served as the base population of the E line to study the influence of long-term selection on the development of nonadditive genetic variation for egg production and body weight traits. Heterosis was significant for BW at 8, 16, and 20 wk of age and at 50% production (females only). At 16 and 20 wk of age, heterosis for BW was significant only for male offspring. No heterosis was observed in the reciprocal crosses for egg production when measured for 84, 180, or 250 d. Heterosis was significant for rate of response to stimulatory lighting of 14 h light per day (days from stimulatory lighting to production of first egg). Based on data for a 250-d production period, heterosis was observed in average clutch length but not in total days lost from broodiness or the effective length of the laying period (250-d lost in periods of 5 or more consecutive d at the end of the laying period). The present results suggest that long-term selection for increased egg production and the correlated decrease in BW increased the relative nonadditive genetic variation in BW. Reciprocal effects were significant for BW at 8 and 16 wk of age, probably due to a large difference in egg weight between the E and RBC1 lines.

Aging↗

PubMed2004

Genetic variation in body weight and egg production in an experimental line selected long term for increased egg production, a commercial dam line, and reciprocal crosses between lines.

A line (E) of turkeys selected long term (40 generations) for increased egg production was reciprocally crossed with a commercial dam line (BD). The BD line was larger and had better conformation than the E line, but the E line laid more eggs than the BD line. Heterosis was negative and significant for BW at 8, 16, and 20 wk of age (both sexes) and at 50% production (females only). Values for heterosis ranged from -3.1 to -5.5%. Negative heterosis was observed for breast width at 16 wk of age (10.8 and 3.0% for males and females, respectively). Positive heterosis values of 9.2, 10.2, and 11.3% were observed for egg production when based on 84, 120, and 180 d, respectively. Heterosis (-8.2%) was significant for rate of response to stimulatory lighting of 14 h light per day (days from stimulatory lighting to production of first egg). Based on data for a 180-d production period, heterosis was observed in average clutch length (12.6%) and rate of lay [number of eggs laid: (180 - total days broody)] (8.2%) but not in total days lost from broodiness or the effective length of the laying period (180 minus days lost in periods of 5 or more consecutive days at the end of the laying period). The present results along with those in the literature suggest that more heterosis in egg production traits might be expected in crosses of lines exhibiting extreme differences in BW and body conformation. The negative heterosis in BW in the BD and E reciprocal crosses reduces the commercial importance of such a cross to improve egg production.

Animals↗

PubMed1987

The effect of PPS on hospital product and productivity.

The results suggest that Prospective Payment System (PPS) prompted a reduction in the proportion of Medicare patients that were discharged, for whom the hospital considered the episode of care to be completed. The results also show a reduction in the proportion of patients discharged dead. When controlling for patient type, the results support the findings, but the magnitude of the change that might be attributed to PPS is somewhat smaller. Proportional changes in the input measures for all patients were next considered. The results indicate that fewer diagnostic tests, fewer laboratory tests, and fewer x-rays were used in 1984. Laboratory tests showed the most dramatic decrease. LOS decreased, but the drug input remained fairly constant. A productivity index that reflects the change in the input measure while controlling for patient type was developed. The results provide strong evidence of a productivity increase in all products for Medicare patients. The drug input did not contribute to the productivity increase. The 50 most frequent DRGs for Medicare patients were examined separately for productivity changes by product. The results further support the findings of an increase in productivity.

Commission on Professional and Hospital Activities↗

PubMed2004

2-aminopurine inhibits lipopolysaccharide-induced nitric oxide production by preventing IFN-beta production.

2-aminopurine (2-AP) is widely used as a specific inhibitor for double stranded-RNA dependent protein kinase (PKR). Here we report that 2-AP can inhibit lipopolysaccharide (LPS)-stimulated nitric oxide (NO) production through the prevention of interferon (IFN)-beta production. 2-AP significantly inhibited NO production in LPS-stimulated RAW 264 murine macrophage cells. 2-AP also reduced the expression of IFN-beta and IFN-inducible genes, such as IFN-gamma-inducible protein (IP)-10 and immune-responsive gene (IRG)-1, and the inducible type of NO synthase (iNOS) mRNA in response to LPS. The addition of exogenous IFN-beta restored 2-AP-inhibited NO production in response to LPS. On the other hand, there was only partial inhibition by 2-AP of nuclear factor (NF)-kappaB activation, IL-6 mRNA expression and tumor necrosis factor (TNF)-alpha production. These results suggested that 2-AP inhibited LPS-induced IFN-beta production by preventing Toll/IL-1 receptor domain-containing adaptor-inducing IFN-beta (TRIF)-dependent signaling rather than myeloid differentiation factor (MyD) 88-dependent signaling, resulting in the inhibition of NO production.

2-Aminopurine↗

PubMed1992

Erythropoiesis in multiple myeloma: defective red cell production due to inappropriate erythropoietin production.

We investigated the pathophysiology of erythropoiesis in 62 patients with multiple myeloma and examined whether it would establish a rational basis for the treatment of their anaemia with recombinant human erythropoietin. Erythropoietin (Epo) production was evaluated by serum levels and erythropoiesis was quantitated by serum transferrin receptor (TfR) levels, both assessed relative to the degree of anaemia. Instead of the expected stimulation of erythropoiesis in response to anaemia, haematocrit correlated positively with marrow erythropoietic activity, indicating that the mechanism of anaemia was primarily defective red cell production. Erythropoiesis decreased and anaemia worsened significantly with advancing clinical stage. 25% of the patients had inadequate Epo production and this proportion increased to 50% in stage 3. Inappropriate Epo production was seen in 60% of patients with renal impairment but was also observed in a number of patients with normal renal function. Erythropoiesis correlated strongly with the adequacy of Epo production, particularly in advanced disease. We conclude that most myeloma patients have defective red cell production even in the absence of massive marrow infiltration and that inappropriate Epo production contributes to their anaemia.

Adult↗

PubMed1990

Cytokine regulation of human monocyte interleukin-1 (IL-1) production in vitro. Enhancement of IL-1 production by interferon (IFN) gamma, tumour necrosis factor-alpha, IL-2 and IL-1, and inhibition by IFN-alpha.

IL-1 production (secreted and cell-associated) was measured in monocyte cultures stimulated by a variety of agents in vitro. Monocytes either adherent to conventional plastic culture plates in serum-free conditions, or in suspension in culture medium containing serum were stimulated to produce IL-1 during culture. In non-adherent, serum-free conditions, monocytes produced very low or undetectable amounts of IL-1 during 20 h of culture. Lipopolysaccharide (LPS) induced equivalent amounts of secreted and cell-associated IL-1, although at very low concentrations more cell-associated IL-1 was produced. IL-1 production in response to LPS could be augmented by crude lymphokine, IFN-gamma, or tumour necrosis factor (TNF) alpha. TNF-alpha preferentially augmented the production of cell-associated IL-1 in LPS-stimulated cultures. TNF-alpha induced a significant amount of IL-1 (mainly cell-associated) directly but could also induce IL-1 secretion when combined with IL-2 or IFN-gamma, or when in the presence of serum. IL-2 acted synergistically with low concentrations of IFN-gamma or IL-1 to induce significant levels of IL-1 production. IFN-alpha did not induce any IL-1 production, but was a potent inhibitor of IL-1 production induced by a variety of stimuli. These results suggest that IL-1 production may be enhanced or reduced by different cytokines at concentrations likely to be found in chronic inflammatory lesions.

Biological Factors↗