Temperature-sensitive regulation of epidermal morphogenesis and the expression of cornified envelope precursors by EGF and TGFα

Epidermis reconstructed on de-epidermized dermis was used to investigate the effects of growth factors and culture temperature on epidermal morphogenesis and the expression of cornified envelope precursors. Cultures grown at 33°C or 37°C in the absence or presence of transforming growth factor alpha (TGFα), keratinocyte growth factor (KGF), basic fibroblast growth factor (bFGF), or insulin-like growth factor (IGF) show a similar morphology to that of native epidermis. Loricrin and SPRR2 are expressed in the stratum granulosum and SPRR3 is absent. Cultures grown in epidermal growth factor (EGF)-supplemented medium at 37°C have a normal morphology, whereas cultures grown at 33°C have a disorganized basal layer, no stratum granulosum, and nuclei are present in the stratum corneum. Loricrin is absent, and SPRR2 and SPRR3 expression extend into the spinous layers. Irrespective of the culture condition used, involucrin is aberrantly expressed in all suprabasal layers. EGF stimulated keratinocyte proliferation and migration to a greater degree than TGFα. Epidermis reconstructed on fibroblast-populated collagen gels at 33°C led to the same disturbances in keratinocyte differentiation as seen in cultures grown on de-epidermized dermis at 33°C in the presence of EGF, whereas parallel cultures grown at 37°C have a similar morphology to that of native epidermis.     

Flexibility of the metabolism of Corynebacterium glutamicum 2262, a glutamic acid-producing bacterium,in response to temperature upshocks

In order to test the temperature sensitivity of glutamate production metabolism, several temperature shifts, from 33 to 37, 38, 39, 40 or 41°C, were applied to the temperature-sensitive strain, Corynebacterium glutamicum 2262, cultivated in a 24-h fed-batch process. Whereas glucose was entirely dedicated to biomass synthesis when cells were grown at 33°C, applying temperature upshocks, whatever their range, triggered a redistribution of the carbon utilisation between glutamate, biomass and lactate production. Although increasing the culture temperature from 33 to 37, 38, 39 or 40°C resulted in final glutamate titers superior to 80 g/l, temperatures resulting in the best chanelling of the carbon flow towards glutamic acid synthesis were 39 and 40°C. Moreover, this study showed that the higher the temperature, the slower the growth rate and the higher the lactate accumulation. Journal of Industrial Microbiology & Biotechnology (2002) 28, 333–337 DOI: 10.1038/sj/jim/7000251 Received 26 September 2001/ Accepted in revised form 23 February 2002     

“Setting paint” analogy for the hydrophobic self‐association of tropoelastin into elastin‐like hydrogel

Alkaline tropoelastin solutions (pH 11) were optically clear at low temperatures, but a firm gel formed when the temperature was raised to 37°C. Reversion to a clear solution took place if the temperature was lowered to below 20°C within less than 2 h, but not if 37°C was maintained for several hours. The precipitated elastin‐like hydrogel thus formed did not visually redissolve at low temperatures. Tropoelastin hydrogel was stable to subsequent washings with alkaline solution at 37°C, but at 4°C some hydrogel redissolved showing that association is at least partly reversible. Washing the hydrogel with neutral 8M urea solution at 4°C dissolved less than 10% of tropoelastin in 24 h. We characterized this phenomenon by combining temperature‐controlled light microscopy analysis, ¹H NMR spectroscopy (temperature, diffusion, and relaxation time studies), and UV‐absorption‐based concentration measurements. The self‐association of tropoelastin at pH 11 is due to hydrophobic interactions in an emulsion‐like system in which the spherules coalesce in a manner like a water‐based latex paint that forms a durable hydrophobic sheet as water and the organic solvent evaporate. In the present case, the sedimentation and entanglement of the tropoelastin porous sheets means that reverse dissolution is a kinetically slow process. © 2009 Wiley Periodicals, Inc. Biopolymers 91: 321–330, 2009. This article was originally published online as an accepted preprint. The “Published Online” date corresponds to the preprint version. You can request a copy of the preprint by emailing the Biopolymers editorial office at biopolymers@wiley.com     

Effect of temperature on the morphometric development of eggs in the prawn Macrobrachium americanum (Caridea: Palaemonidae) and larval success under experimental conditions

Abstract

This study evaluated the effect of temperature on morphometric features of the egg during the embryonic development of the prawn Macrobrachium americanum and the relationship with hatching and the survival of the larvae. Berried females were grouped (n = 3) and reared at three different temperatures, 26, 29, and 33 °C, for which seven developmental stages were recognized. At each stage, the apical and sagittal diameters of the eggs were measured, the volume was calculated, and the weights were recorded. Additionally, the duration of embryonic development, hatching percentage, and larval survival were determined. At 29 and 33 °C, the eggs’ volume increased by 50%, but at 26 °C, the increase was 25%. Larvae from eggs incubated at 33 °C died one day after hatching. At 29 °C, larvae survived until Zoea VII. Larvae from eggs incubated at 26 °C died at the end of Zoea I. The number of days of embryonic development was 20.5 ± 1.5 (26 °C), 15 ± 1 (29 °C), and 12 ± 1 (33 °C). A temperature of 29 °C was the most favorable for embryonic development in M. americanum.     

Temperature management strategy for efficient gene expression in a thermally inducible <Emphasis Type="Italic">Escherichia coli</Emphasis>/bacteriophage system

In a two-phase operation, E. coli containing λSNNU1 (Q ⁻ S ⁻) in the chromosome is typically cultured at 33°C and cloned gene expression is induced by elevating the temperature. At least 40°C is necessary for complete induction of cloned gene expression; however, temperatures above 40°C have been shown to inhibit cloned gene expression. This suggests that a three-phase operation, which has an induction phase between the growth and production phases, may result in higher gene expression. In this study, optimal temperature management strategies were investigated for the three-phase operation of cloned gene expression in thermally inducible E. coli/bacteriophage systems. The optimal temperature for the induction phase was determined to be 40°C. When the temperature of the production stage was 33°C, the optimal time period for the induction phase at 40°C was determined to be 60 min. In contrast, when the temperature of the production phase was 37°C, the optimal period for the induction phase at 40°C was 20∼30 min. When the three-phase temperature and temporal profile were set at a growth phase of 33°C, an induction phase at 40°C for 30 min, and a production phase at 37°C, the highest level of cloned gene expression was achieved.     

Genetic and physiological alterations occurring in a yeast population continuously propagated at increasing temperatures with cell recycling

This work investigated the effects of increasing temperature from 30°C to 47°C on the physiological and genetic characteristics of Saccharomyces cerevisiae strain 63M after continuous fermentation with cell recycling in a system of five reactors in series. Steady state was attained at 30°C, and then the temperature of the system was raised so it ranged from 35°C in the last reactor to 43°C in the first reactor or feeding reactor with a 2°C difference between reactors. After 15 days at steady state, the temperature was raised from 37°C to 45°C for 25 days at steady state, then from 39°C to 47°C for 20 days at steady state. Starter strain 63M was a hybrid strain constructed to have a MAT a/α, LYS/lys, URA/ura genotype. This hybrid yeast showed vigorous growth on plates at 40°C, weak growth at 41°C, positive assimilation of melibiose, positive fermentation of galactose, raffinose and sucrose. Of 156 isolates obtained from this system at the end of the fermentation process, only 17.3% showed the same characteristics as starter strain 63M. Alterations in mating type reaction and in utilization of raffinose, melibiose, and sucrose were identified. Only 1.9% of the isolates lost the ability to grow at 40°C. Isolates showing requirements for lysine and uracil were also obtained. In addition, cell survival was observed at 39–47°C, but no isolates showing growth above 41°C were obtained.     

Observations on the morphology and infectivity for cattle of Babesia bovis parasites in unfed Boophilus microplus larvae after incubation at various temperatures

Dalgliesh R. J. and Stewart N. P. 1979. Observations on the morphology and infectivity for cattle of Babesia bovis parasites in unfed Boophilus microplus larvae after incubation at various temperatures. International Journal for Parasitology9: 115–120. The temperature of incubation of unfed Boophilus microplus larvae infected with Babesia bovis influenced the morphology and infectivity of the Babesia within the tick. Incubation at 37°C for 1–3 days stimulated the development of parasites morphologically similar to those usually observed in fed larvae harvested from cattle; similar forms appeared more slowly in larvae incubated at 31°C or 25°C. Extracts prepared from larvae after incubation at 37°C for 3–5 days or 30°C for 8 days were consistently infective for cattle. Prior storage of larvae at 14°C for up to 28 days enhanced the development of infectivity at 37°C; infectivity could still be produced after 65 days storage at 14°C but not after 76 days. Larvae released on a host transmitted B. bovis sooner if they had been incubated at 37°C for 4 days. It was concluded that the development of B. bovis to an infective stage in B. microplus is temperature dependent and does not require the stimulus of feeding by the host.     

The growth of four species of Azolla as affected by temperature

The growth of 22 strains of Azolla pinnata R. Br., 3 strains of A. filiculoides Lam. and one strain each of A. mexicana Presl and A. caroliniana Willd. was tested separately in liquid culture media kept in controlled, artificial light (30 klux) growth cabinets. Three temperature levels were used: 33°C (37/29°C day/night), 29°C (33/25°C) and 22°C (26/18°C)/ Photoperiod was 12 h a day.For most A. pinnata strains (except three) and an A. mexicana strain the maximum weekly relative growth rate was higher at 33°C than at 22°C, but not for A. filiculoides and A. caroliniana. The highest value of maximum relative growth rate corresponded to 1.9 doubling days and in most strains this occurred in the first week. As the plants grew, the growth rate slowed down more severely at higher temperatures. The maximum biomass was higher at 22°C than at 33°C in all strains. At 22°C, it took 30–50 days to attain maximum biomass and the highest value was 14 g N m^?2 or 320 g dry m^?2 by A. caroliniana, followed by 12 g N m^?2 or 290 g dry wt. m^?2 by one strain of A. filiculoides. At 29°C, the maximum biomass was attained in 20–35 days. The highest value was 6.3 g N m^?2 or 154 g dry wt. m^?2 by A. caroliniana. At 33°C, most A. pinnata strains gave a maximum biomass of less than 4 g N m^?2 after 13–23 days, while some strains grew up to 30 days, resulting in a higher maximum biomass. The highest maximum biomass at 33°C was 5.5 g N m^?2 or 140 g m^?2 dry wt. by A. pinnata from Cheng Mai while the maximum biomass of A. filiculoides and A. caroliniana was much less. Azolla filiculoides requires lower temperature than other species for its growth. Azolla pinnata has the best tolerance to high temperatures among the four species. Azolla mexicana could not be discriminated from A. pinnata in its response to temperature. Azolla caroliniana may keep an intermediate position between A. filiculoides and A. pinnata in temperature response.The formation of ammonia in the medium was examined and it occurred mostly under stationary growth conditions, but, at 33°C, some strains of A. pinnata and A. mexicana released or formed ammonia at 0.3–0.8 μg N ml^?1 per week during their initial exponential growth stage.     

Thermotolerance and thermal sensitization in a differentiating murine erythroleukemia cell line

• 1.1.|Friend erythroleukemia cells (FELC, a differentiating cell line) were heated at various temperatures and heating sequences. Heat treatments which ranged from 41.0 to 45.0°C and did not cause differentiation in FELC and inhibited the differentiation response to DMSO in FELC.
• 2.2.|Heating resulted in cell killing which increased with temperature and heating time. Protracted low temperature heating (40.0–42.0°C) or incubation at 37°C between two heat treatments at 45.0°C resulted in thermotolerance for both the endpoints of cell killing and differentiation.
• 3.3.|High temperature heating (45.0°C) before heating at 41.0–42.0°C resulted in increased thermal sensitivity to the latter heat treatments. This was observed for both the survival and differentiation endpoints.
• 4.4.|A comparison was made of the thermal sensitivity for the two endpoints of cell killing and differentiation.

     

Influence of different temperatures on the tachinid parastite,Sturmiopsis inferens [Dip.]

The influence of constant temperatures of 27, 29, 31 and 33°C and alternating temperature of 31/33°C (18/6 h) onSturmiopsis inferens Townsend was studied during 12 successive generations. The larval and pupal periods for male parasites were 13.5±0.5 and 11.0±0.3 days respectively and for female 12.8±0.5 and 11.1±0.3 days respectively in the 1st generatioin at 27°C. It decreased progressively with increase in temperature. Survival of females, fertility and fecundity were adversely affected at higher temperatures. A temperature range of 27–29°C appeared to be optimum for mass rearing of the parasite in the laboratory. The higher premature mortality observed at a constant 33°C was not observed at temperatures fluctuating between 31/33°C. Presumably under field conditions, where temperature is constantly fluctuating, the flies will be able to withstand a comparatively higher temperature.     

Cell-mediated mineralization in culture at low temperature associated with subtle thermogenic response

In both the growth plate and in marrow stromal cell cultures cell-mediated mineralization is preceded by characteristics of anaerobic and low efficiency energy metabolism. Reagents that increase mineralization like malonate and dexamethasone (DEX) also increase the mitochondrial membrane potential (MtMP) especially 1 week after DEX stimulation. Contrarily, levamisole, which decreases mineralization, also decreases MtMP. Modulation of MtMP and energy metabolism could be linked to regulation of mineralization by the uncoupling of oxidative phosphorylation. This uncoupling should be associated with thermogenesis in cells that induce mineralization. We examined whether cold temperature affects mineralization, and whether cellular thermogenesis takes place at cold temperature in parallel to changes in MtMP. Osteoprogenitor cells (OPC) induced, in DEX stimulated rat marrow stroma, higher mineralization at 33°C than at 37°C. Increased mineralization by cold temperature required long incubation since incubation in the cold during short intervals, 3–4 days, did not increase mineralization relative to (37°C) controls. Marrow stromal cells in the presence of valinomycin responded to incubation at 33°C by retaining all the vital dye after 4 h, unlike the cells at 37°C; however, after 24 h the level of dye retention at 33°C was the same as at 37°C. The delayed response of the temperature-dependent (> 37°C) K⁺ ionophor to incubation in the cold indicated that certain cells may respond to low temperature by local intracellular heating, and by heat conduction to the plasma membrane. DEX-stimulated stromal cells, unlike unstimulated cells, showed increased mitochondrial rhodamine 123 retention in the presence of valinomycin after 24 h in the cold, which corresponds to day 4 of OPC induction. This is consistent with the concept that valinomycin-induced cell damage is mediated by (cold-induced) local heating. The mechanism of this cell damage should selectively prefer non-thermogenic (rhodamine retaining) over thermogenic (rhodamine leaking) cells such as OPC. At cold temperature DEX-stimulated stromal cells showed the best anti-OPC selection under exposure to valinomycine between days 3–7, concurrent with the period of rhodamine leakage from the mitochondria. These results indicate that thermogenesis is enhanced during the period of low MtMP in mineralizing cells, and prolonged exposure to cold increases mineralization also due to induction of subtle thermogenesis. © 1996 Wiley-Liss, Inc.     

Dynamic Light Scattering Studies on Hydrodynamic Properties of Fibrinogen-Fibronectin Complex

Abstract

A high molecular weight ‘cryogel’ was obtained as insoluble complexes by cold incubation at near-freezing temperatures from heparinized plasma of patients with rheumatoid arthritis. After the cryogel was solubilized at 37°C, 1:1 complex of fibrinogen and fibronectin was purified at room temperature by affinity chromatography on a gelatin-Sepharose 4B. Hydrodynamic properties of the complex were investigated as a function of temperature and NaCl concentration using a dynamic light scattering. The diffusion coefficients of the complex at 20°C decreased with increasing of NaCl concentration as free fibronectin. The complex appears to be a more compact form at low ionic concentration, which is associated with conformational changes of fibronectin. The diffusion coefficient of the complex at 20°C in 0.05 M Tris- HCl(pH7.4) containing 0.5 M NaCl was estimated as 8.5× 10^?8 cm²s^?1. The complex did not dissociate over the temperature range from 20 to 37°C. The diffusion coefficients of the complex decreased significantly at 12°C and 40°C. The thermal denaturation of fibrinogen molecule in the complex was observed at 40°C. The CONTIN analysis of the light scattering data showed that the complex associated to form higher aggregates at 15°C, but not at near- freezing temperature. The equilibrium between the complex and higher aggregates appeared reversible.     

Determination of temperature sensitive stages for sexual differentiation of the gonads in embryos of the turtle,Emys orbicularis

In order to determine the temperature sensitive stages for sexual differentiation of the gonads in Emys orbicularis, eggs of this turtle were shifted at different stages of embryonic development from the male-producing temperature of 25°C to the female-producing temperature of 30°C and reciprocally. Based on the series of developmental stages described by Yntema (′68) for Chelydra serpentina, temperature begins to influence sexual differentiation of Emys orbicularis at stage 16, a stage in which the gonads are still histologically undifferentiated. Its action lasts over the first steps of histological differentiation of the gonads. The minimal exposure at 25°C required for male differentiation of all individuals extends from stage 16 to somewhat before stage 21. For 100% female differentiation, incubation at 30°C must be longer, from stage 16 to somewhat before stage 22. Shorter exposures at 25°C or 30°C during these periods result in different percentages of males, females, and intersexes. Our results show that there is a critical stage (stage 16) which is the same for both male and female differentiation of the gonads. The thermosensensitive periods are rather long, corresponding to 11–12 days at 25°C and 30°C.     

Effects of temperature on biology and life table parameters of the Asian citrus psyllid, Diaphorina citri Kuwayama (Homoptera: Psyllidae)

The development, survivorship, longevity, reproduction, and life table parameters of the Asian citrus psyllid, Diaphorina citri Kuwayama were evaluated at 10°C, 15°C, 20°C, 25°C, 28°C, 30°C and 33°C. The populations reared at 10°C and 33°C failed to develop. Between 15°C and 30°C, mean developmental period from egg to adult varied from 49.3 days at 15°C to 14.1 days at 28°C. The low‐temperature developmental thresholds for 1st through 5th instars were estimated at 11.7°C, 10.7°C, 10.1°C, 10.5°C and 10.9°C, respectively. A modified Logan model was used to describe the relationship between developmental rate and temperature. The survival of the 3rd through 5th nymphal instars at 15–28°C was essentially the same. The mean longevity of females increased with decreasing temperature within 15–30°C. The maximal longevity of individual females was recorded 117, 60, 56, 52 and 51 days at 15°C, 20°C, 25°C, 28°C and 30°C, respectively. The average number of eggs produced per female significantly increased with increasing temperature and reached a maximum of 748.3 eggs at 28°C (P<0.001). The population reared at 28°C had the highest intrinsic rate of increased (0.199) and net reproductive rate (292.2); and the shortest population doubling time (3.5 days) and mean generation time (28.6 days) compared with populations reared at 15–25°C. The optimum range of temperatures for D. citri population growth was 25–28°C.     

Pyrimidine Deoxyribonucleosides Are Phosphorylated to Triphosphates at Low Temperatures Too,But Are Not Incorporated into DNA or Phospholipids.

Abstract

Thymidine (Thd) was phosphorylated to dTTP also at 0°C, both in Ehrlich ascites tumor cells and human tonsillar lymphocytes, but was not incorporated into DNA. The uptake and phosphorylation of ¹⁴C-Thd into the pool showed regular kinetics (Km 6, 6 uM), and the main metabolite was dTTP (75–84%) both at 0°C, and 37°C. Similarly, deoxycytidine (dCyd) was also transported and phosphorylated to nucleotides (76%) at low temperature, but no incorporation into DNA and phospholipid precursor liponucleotides could be detected at 0°C. Under the same conditions, at 37°C, when lymphocytes were labeled with 5-³H-dCyd, 51% of the total pool radioactivity was found in liponucleotides. Transport and phosphorylation of deoxynucleosides seem to be tightly coordinated at both temperatures, which processes are directly coupled to membrane-phospholipid and DNA biosynthesis, but only at physiological temperature while they seem “uncoupled” at low temperature. The fact that nucleoside phosphorylation occures also at low temperature has implications for several experimental techniques used in cell biology.     

Effects of temperature on the development and fecundity of Microplitis similis (Hymenoptera: Braconidae), a parasitoid of Spodoptera litura (Lepidoptera: Noctuidae)

Microplitis similis (Hymenoptera: Braconidae) is a solitary endoparasitoid of Spodoptera litura larvae (Lepidoptera: Noctuidae). Here, the effects of constant temperature (18, 21, 24, 27, 30, 33 and 36 °C) on the development and fecundity of M. similis developing in S. litura were studied in the laboratory to clarify the range of its potential distribution and better understand its potential as a biological control agent. The developmental duration of M. similis varied from 10.6 (33 °C) to 27.9 days (18 °C). The developmental threshold temperature and effective accumulative temperature of M. similis were 9.96 °C and 231.14 Degree-days, respectively. The average adult longevity of M. similis ranged from 5.1 (33 °C) to 26.8 days (18 °C). The maximum fecundity of the parasitoid was observed at 27 and 30 °C, which were 43.07 and 39.73 eggs, respectively. The minimum fecundity of the parasitoid was observed at 18 °C, which was 8.27 eggs. The intrinsic rate of increase (r_m) and finite rate of increase (λ) of M. similis were the highest at 30 °C. The net reproduction rate (R₀) was the highest at 27 °C and 30 °C, which were 44.34 and 40.39, respectively. We concluded that temperatures in the range 27–30 °C are the most suitable for development and reproduction of M. similis. Our study provides detailed basic information for development and reproduction of M. similis under different temperature conditions.     

Comparative growth and metabolism of gelatinous colonies of three cyanobacteria,Nostoc commune,Nostoc pruniforme and Nostoc zetterstedtii,at different temperatures

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Temperature-Induced Catch-Slip to Slip Bond Transit in Plasmodium falciparum-Infected Erythrocytes

Plasmodium falciparum malaria-infected red blood cells (IRBCs), or erythrocytes, avoid splenic clearance by adhering to host endothelium. Upregulation of endothelial receptors intercellular adhesion molecule-1 (ICAM-1) and cluster of differentiation 36 (CD36) are associated with severe disease pathology. Most in vitro studies of IRBCs interacting with these molecules were conducted at room temperature. However, as IRBCs are exposed to temperature variations between 37°C (body temperature) and 41°C (febrile temperature) in the host, it is important to understand IRBC-receptor interactions at these physiologically relevant temperatures. Here, we probe IRBC interactions against ICAM-1 and CD36 at 37 and 41°C. Single bond force-clamp spectroscopy is used to determine the bond dissociation rates and hence, unravel the nature of the IRBC-receptor interaction. The association rates are also extracted from a multiple bond flow assay using a cellular stochastic model. Surprisingly, IRBC-ICAM-1 bond transits from a catch-slip bond at 37°C toward a slip bond at 41°C. Moreover, binding affinities of both IRBC-ICAM-1 and IRBC-CD36 decrease as the temperature rises from 37 to 41°C. This study highlights the significance of examining receptor-ligand interactions at physiologically relevant temperatures and reveals biophysical insight into the temperature dependence of P. falciparum malaria cytoadherent bonds.     

A novel low-temperature-active β-glucosidase from symbiotic <Emphasis Type="Italic">Serratia</Emphasis> sp. TN49 reveals four essential positions for substrate accommodation

To maximize the production of flag-tagged cartilage oligomeric matrix protein angiopoietin-1 (FCA1) from Chinese hamster ovary (CHO) cells, the effects of culture pH and temperature on cell growth and FCA1 production were investigated. Cells were cultivated in a bioreactor at different culture pH (6.7, 6.9, 7.2, and 7.5) and temperatures (33 and 37 °C). Lowering the culture temperature suppressed cell growth while allowing maintenance of high cell viability for a longer culture period. The specific FCA1 productivity (q _FCA1) was increased at low culture temperature. Accordingly, the highest FCA1 concentration was obtained at pH 7.2 and 33 °C, and was approximately 4.0-fold higher than that at pH 7.2 and 37 °C. However, aggregates and a monomeric form of FCA1, which are undesirable due to reduced biological activity or immunogenicity, were significant at pH 7.2 and 33 °C. It was also found that the expression pattern of FCA1 was affected more significantly by culture pH than by the culture temperature. FCA1 aggregation dramatically decreased at culture pH 7.5 regardless of the culture temperature. Furthermore, the monomeric form of FCA1 was not observed. Taken together, optimization of culture temperature and culture pH (33 °C and pH 7.5) significantly improves the production of biologically active FCA1 with tetrameric or pentameric forms from CHO cells.