Cell cycle model for growth rate and death rate in continuous suspension hybridoma cultures

As a result of recent advances in flow cytometry, renewed interest is shown in modeling the kinetic behavior of cells in culture on the basis of cell cycle parameters. An important but often overlooked kinetic variable in hybridoma cultures is the cell death rate. Not only the overall cell growth but also the kinetics of nutrient metabolism and monoclonal antibody production have been shown to depend on the cell death rate in continuous suspension hybridoma cultures. The present study shows that the death rate in hybridoma cultures is proportional to the fraction of cells arrested in the G(1) phase of the cell cycle. The steady-state cell age distributions in the various phases of the division cycle have been calculated analytically. A simple mathematical model has been used to produce the profiles of the cycling and arrested cell fractions with respect to the dilution rate. The calculated steady-state growth rate, death rate, and viability profiles are shown to be in agreement with recently published experimental data from continuous suspension hybridoma cultures. (c) 1992 John Wiley & Sons, Inc.     

Effects of electroconductive heat treatment and electrical pretreatment on thermal death kinetics of selected microorganisms

Suspensions of yeast cell (zygo Saccharomyces bailii) in a phosphate buffer solution were subjected to conventional (hot water) and ohmic (electric current) heating under identical temperature histories. Experiments were also conducted with cells of Escherichia coli to compare the lethal effect of combination of sublethal electrical preteatment and conventional heating with conventional heating. The kinetic parameters (D,Z,K and E(a)) were determined for both organisms during different treatments. There was no significant difference in the death rate of yeast cells during conventional and ohmic heating at the voltage range used in this study. Results of electrical pretreatment and conventional heating on E. coli indicated differences under certain conditions when compared with pure conventional heating. Thus it is concluded that microbial death during ohmic heating was due primarily to thermal effects with no significant effect of electric current per se. Sublethal electrical pretreatment appears to offer potential for increased bacterial inactivation in certain cases.     

The thermal and energetic significance of clustering in the speckled mousebird,Colius striatus

Summary The effect of clustering behaviour on metabolism, body temperature, thermal conductance and evaporative water loss was investigated in speckled mousebirds at temperatures between 5 and 36°C. Within the thermal neutral zone (approximately 30–35 °C) basal metabolic rate of clusters of two birds (32.5 J·g^-1·h^-1) and four birds (28.5 J·g^-1·h^-1) was significantly lower by about 11% and 22%, respectively, than that of individuals (36.4 J·g^-1·h^-1). Similarly, below the lower critical temperature, the metabolism of clusters of two and four birds was about 14% and 31% lower, respectively, than for individual birds as a result of significantly lower total thermal conductance in clustered birds. Body temperature ranged from about 36 to 41°C and was positively correlated with ambient temperature in both individuals and clusters, but was less variable in clusters. Total evaporative water loss was similar in individuals and clusters and averaged 5–6% of body weight per day below 30°C in individuals and below 25°C in clusters. Above these temperatures total evaporative water loss increased and mousebirds could dissipate between 80 and 90% of their metabolic heat production at ambient temperatures between 36 and 39°C. Mousebirds not only clustered to sleep between sunset and sunrise but were also observed to cluster during the day, even at high ambient temperature. Whereas clustering at night and during cold, wet weather serves a thermoregulatory function, in that it allows the brrds to maintain body temperature at a reduced metabolic cost, clustering during the day is probably related to maintenance of social bonds within the flock.Abbreviations BMR basal metabolic rate - bw body weight - C _totab total thermal conductance - EWI evaporative water loss - M metabolism - RH relative humidity - T _a ambient temperature - T _b body temperature - T _ch chamber temperature - T _cl cluster temperature - TEWL total evaporative water loss - LCT lower critical temperature - TNZ thermal neutral zone     

Killer polyamines?

Mammalian cells can rapidly make large changes in their rate of polyamine biosynthesis in response to mitogenic and trophic signals. However, cultured cells seem to grow adequately as long as they are supplied a steady but unregulated supply of polyamines. This implies that complex and rapid changes in polyamine synthesis serve a function in a special rather than a general biological context. We suggest that the appropriate context in which regulation of polyamines mediates crucial functions is the mammalian embryo and that one function of polyamines is to act as substrate in an oxidative pathway that arbitrates programmed cell death.     

Thermal acclimation and hardening in tadpoles of the bullfrog, Rana catesbeiana

1. 1.|Critical thermal maxima (CTMax) and minima (CTMin) were measured to evalute thermal hardening in Rana catesbeiana.

2. 2.|Tadpoles show heat hardening and CTMax acclimation, and both responses are influenced by developmental stage.

3. 3.|The first evidence of cold hardening in vertebrates is reported here.

4. 4.|Heat hardening significantly reduces cold tolerance, but there is otherwise no evidence of a cross-hardening effect.

Oligouridylates as a template for nonenzymatic synthesis of oligoadenylates

Summary Chemical polymerization of adenosine-5-phosphorimidazolide was conducted in the presence of oligouridylate templates. Oligo U with chain length more than eight served as a template and facilitated oligoadenylate formation. No template activity was observed when oligo U up to a hexamer was used. These results correlate with thermal transition temperatures of oligo U-pA complexes.     

Thermal Denaturation of Native Striatal Tyrosine Hydroxylase: Increased Thermolability of the Phosphorylated Form of the Enzyme

Tyrosine hydroxylase was purified from bovine corpus striatum. The native enzyme had a half-life of 15 +/- 3 min at 50 degrees C. Phosphorylation of tyrosine hydroxylase with protein kinase purified from both corpus striatum and heart activated the enzyme, but activity was rapidly lost with additional preincubation of the enzyme at 30 degrees C. Thermal denaturation studies indicated that phosphorylated tyrosine hydroxylase had a half-life of 5 +/- 2 min at 50 degrees C     

Immobilization of luciferase from the firefly Luciola mingrelica — Catalytic properties and stability of the immobilized enzyme

Firefly (Luciola mingrelica) luciferase [Photinus luciferin 4-monooxygenase (ATP-hydrolysing); Photinus luciferin: oxygen 4-oxidoreductase (decarboxylating, ATP-hydrolysing), EC 1.13.12.7] has been immobilized on albumin and polyacrylamide gel, on AH-, CH- and CNBr-Sepharose 4B as well as on Ultragel, Ultradex and cellophane film activated by cyanogen bromide. Only immobilization on cyanogen bromide-activated polysaccharide carriers resulted in highly active immobilized luciferase. Kinetic properties of immobilized luciferase hardly differed from those of the soluble enzyme. The inactivation rate constants of soluble and immobilized luciferase were measured at pH 5.5–9.0 and 25°C as well as at pH 7.8 and 20–40°C. The ΔH^≠ and ΔS^≠ values for inactivation of soluble and immobilized luciferases were obtained. A 1000-fold stabilization effect was noted for the luciferase immobilized on CNBr-Sepharose 4B at pH 7.5 and 25°C. A stabilization mechanism for the immobilized luciferase is discussed.     

A laboratory study on the thermal tolerance of four southeastern stream insect species (Trichoptera,Ephemeroptera)

The acute thermal tolerances of four southeastern stream insect species, Ephemerella invaria (Walker), Stenonema ithaca (Clemens and Leonard), Symphitopsyche morosa (Hagun), and Brachycentrus lateralis (Say) were determined using an artificial stream enclosure. All species were acclimated at 10°C for 72 hours prior to instantaneous immersion into heated water for 96 hours. Percent mortality was recorded and the temperature at which 50% mortality occurred determined (LT5o). Data were subjected to standard statistical analysis.Thermal tolerance values were compared between species tested and to results from previous investigations using similar methodologies. The evolution and life histories of these species were also discussed in relation to their thermal tolerance values.