Effect of Bcl‐xL overexpression on lactate metabolism in chinese hamster ovary cells producing antibody

Previously, overexpression of anti‐apoptotic proteins, such as E1B‐19K and Aven, was reported to alter lactate metabolism of CHO cells in culture. To investigate the effect of Bcl‐x_L, a well‐known anti‐apoptotic protein, on lactate metabolism of recombinant CHO (rCHO) cells, two antibody‐producing rCHO cell lines with regulated Bcl‐x_L overexpression (CS13*‐0.02‐off‐Bcl‐x_L and CS13*‐1.00‐off‐Bcl‐x_L) were established using the Tet‐off system. When cells were cultivated without Bcl‐x_L overexpression, the specific lactate production rate (q_Lac) of CS13*‐0.02‐off‐Bcl‐x_L and CS13*‐1.00‐off‐Bcl‐x_L were 7.32 ± 0.37 and 6.78 ± 0.56 pmol/cell/day, respectively. Bcl‐x_L overexpression, in the absence of doxycycline, did not affect the q_Lac of either cell line, though it enhanced the viability during cultures. Furthermore, activities of the enzymes related to glucose and lactate metabolism, such as hexokinase, glucose‐6‐phosphate dehydrogenase, lactate dehydrogenases, and alanine aminotransferase, were not affected by Bcl‐x_L overexpression either. Taken together, Bcl‐x_L overexpression showed no significant effect on the lactate metabolism of rCHO cells. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:1594–1598, 2013     

Influence of pluronic F68 on oxygen mass transfer

Pluronic F68 is one of the most used shear protecting additives in cell culture cultivations. It is well known from literature that such surface‐active surfactants lower the surface tension at the gas‐liquid interface, which influences the mass transfer. In this study, the effect of Pluronic F68 on oxygen mass transfer in aqueous solutions was examined. Therefore, the gassing in/gassing out method and bubble size measurements were used. At low concentrations of 0.02 g/L, a 50% reduction on mass transfer was observed for all tested spargers and working conditions. An explanation of the observed effects by means of Higbie's penetration or Dankwerts surface renewal theory was applied. It could be demonstrated that the suppressed movement of the bubble surface layer is the main cause for the significant drop down of the k_La‐values. For Pluronic F68 concentrations above 0.1 g/L, it was observed that it comes to changes in bubble appearance and bubble size strongly dependent on the sparger type. By using the bubble size measurement data, it could be shown that only small changes in mass transfer coefficient (k_L) take place above the critical micelle concentration. Further changes on overall mass transfer at higher Pluronic F68 concentrations are mainly based on increasing of gas holdup and, more importantly, by increasing of the surface area available for mass transfer. © 2013 American Institute of Chemical Engineers Biotechnol. Prog., 29:1278–1288, 2013     

Photosynthetic adaptation of Euphorbia fractiflexa (Euphorbiaceae) and survival in arid regions of the Arabian Peninsula

Arid regions of the Arabian Peninsula are characterized by a short wet season with erratic rainfall, high temperature, and high evaporation. In these arid regions, the leafless stem succulent Euphorbia fractiflexa S. Carter & J.R.I.Wood (Euphorbiaceae) is an abundant perennial. Work presented in this paper aimed at investigating crassulacean acid metabolism (CAM) as a physiological adaptation crucial for survival of E. fractiflexa in arid regions of the Arabian Peninsula. Work involved investigations of stomatal diffusive conductance, chlorophyll fluorescence, and cell sap titratable acidity. Results represent the first report of obligate CAM in E. fractiflexa. Low values of stomatal conductance and dampening of CAM acidification–deacidification cycles during the long dry season also denoted tendency of this species to shift from obligate CAM to CAM-idling. Results also showed water stress-induced reduction in Photosystem II (PSII) activity occurring in concomitance with increased non-photochemical quenching of chlorophyll fluorescence denoting operation of non-photochemical energy dissipation mechanisms that are important for photoprotection of the photosynthetic machinery under stress conditions.     

Dissecting the energy metabolism in Mycoplasma pneumoniae through genome‐scale metabolic modeling

Mycoplasma pneumoniae, a threatening pathogen with a minimal genome, is a model organism for bacterial systems biology for which substantial experimental information is available. With the goal of understanding the complex interactions underlying its metabolism, we analyzed and characterized the metabolic network of M. pneumoniae in great detail, integrating data from different omics analyses under a range of conditions into a constraint‐based model backbone. Iterating model predictions, hypothesis generation, experimental testing, and model refinement, we accurately curated the network and quantitatively explored the energy metabolism. In contrast to other bacteria, M. pneumoniae uses most of its energy for maintenance tasks instead of growth. We show that in highly linear networks the prediction of flux distributions for different growth times allows analysis of time‐dependent changes, albeit using a static model. By performing an in silico knock‐out study as well as analyzing flux distributions in single and double mutant phenotypes, we demonstrated that the model accurately represents the metabolism of M. pneumoniae. The experimentally validated model provides a solid basis for understanding its metabolic regulatory mechanisms.     

Respiratory control in aquatic insects dictates their vulnerability to global warming

Forecasting species responses to climatic warming requires knowledge of how temperature impacts may be exacerbated by other environmental stressors, hypoxia being a principal example in aquatic systems. Both stressors could interact directly as temperature affects both oxygen bioavailability and ectotherm oxygen demand. Insufficient oxygen has been shown to limit thermal tolerance in several aquatic ectotherms, although, the generality of this mechanism has been challenged for tracheated arthropods. Comparing species pairs spanning four different insect orders, we demonstrate that oxygen can indeed limit thermal tolerance in tracheates. Species that were poor at regulating oxygen uptake were consistently more vulnerable to the synergistic effects of warming and hypoxia, demonstrating the importance of respiratory control in setting thermal tolerance limits.     

Temperaturverlauf und Gärfutterqualität unter verschiedenen Silodecken im Flachsilo

Observations were carried out of actual acidity, volatile fatty acid (VFA) concentrations, enzyme activity in the rumen, total protein, urea, total lipid and glucose in the serum of conventional (CL) and gnotobi‐otic lambs (GL) in the period of milk nutrition. The inoculum of gnotobiotic iambs contained Streptococcus bovis, Prevoxella ruminicola, Butyrivibrio fibrisolvens and Selenomonas ruminantium at a concentration of 1.10⁶ each. Throughout the observation period the pH of the rumen contents of gnotobiotic lambs ranged within 6.5–6.8 with a significant difference at an age of 7 weeks. Total VFA concentrations in the rumen contents were increased in the CL throughout milk nutrition; the differences at 4 and 5 weeks of age were significant. Total VFA in the conventional lambs revealed an increasing tendency between weeks 4 and 7, reaching higher levels at 7 weeks of age (57.1 mmol.l"¹), whereas in the gnotobiotic animals the range (24.3–30.1 mmol.l"¹) was narrow and the peak occurred at 6 weeks of age. In GL significantly increased molar proportions of acetic acid were observed whereas in CL the molar proportions of propionic acid proved to be significant increased. The molar proportions of butyric and valeric acids were increased in CL but the group differences were not significant. In GL no isoacids were found. Alpha amylase (E.C.3.2.1.1.) activity of the rumen contents was significantly increased in GL between weeks 2 and 6 of age whereas cel‐Iulase (endoglucanase E.C.3.2.1.4. and cellobiohydrolase E.C.3.2.1.91.) activity was significantly increased in 4‐week‐old CL. Over the whole period of milk nutrition no significant differences were observed in ure‐ase (E.C.3.5.1.5.) activity of the rumen contents in the examined groups.

At 5 weeks of age significantly increased total protein levels were observed in the conventional animals with maximum levels occurring at 4 weeks of age (CL‐59.5 g.l"¹ GL‐55.3 g.l"¹). Urea levels in 6‐week old conventional lambs were significantly higher than in the gnotobiotic animals (CL‐6.4 mmol.l"’ vs. GL‐1.9 mmol.l"¹). As to glycaemia no significant group differences were recorded. In the conventional animals total lipid levels were significantly increased at 1 and 6 weeks of age with a peak occurring in the first week of life (7.5 g.l"¹) whereas in the gnotobiotic lambs a significant increase was observed at 3 weeks of age, the peak being recorded in 4 week‐old animals (4.3 g.l"¹). Throughout the period of interest the mean daily weight gains in the conventional and gnotobiotic lambs presented 0.164 and 0.162 kg, respectively. The functional development of the rumen was significantly influenced by the complexity of the microfiora.     

Chemische Untersuchungen über unterschiedlich vorbehandelte Federmehle

Abstract

Dr. Carlheinrich Engelmann: Ernährung und Fütterung des Geflügels. Erschienen im Neumann-Verlag Radebeul/Berlin, 162 Seiten, 17 × 24 cm Halbleinen, MDN 6,80. Reviewed by H. Bergner

Dr. Martin Pankrath: Wasserbedarf landwirtschaftlicher Nutztiere — Mengennormen. - Erschienen im S. Hirzel Verlag, Leipzig C 1, Schuhmachergäßchen 1–3. 1. Auflage 1963, 99 Seiten mit 25 Diagrammen, 21 Tabellen und 2 Tafeln im Text. 8°, Hln., MDN 10,70. Reviewed by H. Bergner

Physiologische und wirtschaftliche Leistungsgrenzen in der Tierzucht 14 Vorträge eines internationalen Symposions im Max-Planck-Institut für Tierzucht und Tierernährung in Mariensee vom 14. bis 17. Oktober 1963.: Herausgegeben von M. Witt. 315 Seiten mit 8 Kunstdrucktafeln. Englische Summaries. Steif broschiert, Preis DM 16,-. Bestellung über Institut. Reviewed by H. Bergner     

Untersuchungen über die Wirkung von Vitamin B12 und behandeltem Sojaschrot bei der Mast von Küken

To examine the effects of dietary conjugated linoleic acids (CLA) on lipid metabolism and antioxidant capacity in laying hens, Hy-Line Brown layers (n = 384, 52 weeks old) were randomly allocated to one of four dietary treatments. Each treatment had six replicates of 16 hens each. All birds were assigned to acorn-soybean meal-based diet containing a mixture of CLA at 0%, 1%, 2% or 4% for six weeks. With increasing dietary CLA, egg weight and feed intake decreased, and yolk colour was darkened. Feed efficiency was improved at 1% and 2% dietary CLA. Serum triglyceride concentration was significantly reduced by CLA in a dose dependent manner. A linear decrease in total cholesterol and low-density lipoprotein cholesterol, and an increase in high-density lipoprotein cholesterol levels were observed after CLA supplementation. With increasing dietary CLA, the deposition of two major isomers of CLA (c9, t11; t10, c12) in yolk lipids increased linearly, the proportion of saturated fatty acids increased and monounsaturated fatty acids decreased significantly. The proportion of polyunsaturated fatty acids was highest at 1% CLA. Compared to the control, CLA supplementation significantly increased the activities of superoxide dismutase and glutathione peroxidase, inhibited hydroxyl radicals and superoxide anion production, and decreased the malonaldehyde concentrations in both serum and liver. The results demonstrated that dietary CLA meliorated serum lipid profiles and enhanced the antioxidant capacity of laying hens.     

Untersuchungen zur Frage der Methionintoxizität

The study aimed to examine effects of supplemented conjugated linoleic acids (CLA) to periparturient cows receiving different concentrate proportions ante partum (a.p.) to investigate CLA effects on lipid mobilisation and metabolism. Compared to adapted feeding, a high-concentrate diet a.p. should induce a ketogenic metabolic situation post partum (p.p.) to better understand how CLA works. Sixty-four pregnant German Holstein cows had ad libitum access to partial mixed rations 3 weeks prior to calving until day 60 p.p. Ante partum, cows received control fat (CON) or a CLA supplement at 100 g/d, either in a low-concentrate (CON-20, CLA-20) or high-concentrate diet (CON-60, CLA-60). Post partum, concentrate proportion was adjusted, while fat supplementation continued. After day 32 p.p., half of the animals of CLA-groups changed to CON supplementation (CLA-20-CON, CLA-60-CON). A ketogenic metabolic situation p.p. was not achieved and therefore impacts of CLA could not be examined. Live weight, milk yield and composition, blood parameters remained unaffected by the treatments. Only a slightly reduced milk fat yield (not significant) was recorded for Group CLA-20. The proportion of trans-10,cis-12 (t10,c12) CLA in milk fat was significantly increased in CLA-groups compared to CON-groups. With the exception of a reversible CLA effect on milk fat in Group CLA-20, no post-treatment effects occurred. Dry matter intake (DMI) of Group CLA-60 was highest before calving, resulting in a significantly improved estimated energy balance after calving. Ante partum, net energy intakes were significantly increased in high-concentrate groups. Overall, supplemented CLA preparation did not relieve metabolism and lipid mobilisation of early lactating cows. But feeding CLA in a high-concentrate diet a.p. seems to increase DMI and thereby improve the energy balance of cows immediately after calving.