Constancy of proline hydroxylation in collagen synthesized at different temperatures by poikilotherms.

Previous experiments have suggested that the body temperature of an organism may in part regulate the hydroxyproline content of its collagen. In order to test this hypothesis, the hydroxyproline content of the collagen synthesized at different temperatures by fish cells in culture and by intact amphibian larvae was measured. High levels of prolyl hydroxylation were found at both low and high temperatures, which eliminates the possibility that temperature can directly modulate prolyl hydroxylation in vivo. Synthesis and hydroxylation of collagen appear to be coordinated so that unstable underhydroxylated molecules are not produced under normal circumstances.     

Competition between two planktonic rotifer species at different temperatures: an experimental test

1. The effect of temperature on the outcome of resource competition between two planktonic rotifers (Synchaeta pectinata and Brachionus calyciflorus) was investigated in laboratory experiments. In addition to the competition experiments, several physiological variables and their temperature‐dependence were characterised, including ingestion rate and starvation tolerance. 2. Because of a lower threshold food level (TFL) for population growth for the food algae Cryptomonas erosa, Synchaeta was predicted to be the superior competitor at low temperatures (12 °C). In contrast, Brachionus had a lower TFL at 20 °C and was predicted to be competitively superior at this temperature. 3. In both rotifer species, ingestion rates increased with temperature, but the increase was much more pronounced in Brachionus. Ingestion rates of Brachionus at temperatures from 8 to 24 °C were always higher than in Synchaeta (up to 4.6‐fold). 4. Starvation resistance reduced with temperature in both rotifer species. At all temperatures investigated (12, 16 and 20 °C) Brachionus could survive starvation for longer than Synchaeta. This difference was strongest at 12 °C (5.8 days versus 2.5 days). 5. In the first competition experiment, food was supplied at 48 h‐intervals. Brachionus displaced Synchaeta at both experimental temperatures (12 and 20 °C). Competitive exclusion of Synchaeta at the lower temperature was probably because of large fluctuations in algal densities that resulted from the long intervals between feeding, a condition that favoured Brachionus because of its higher starvation resistance. 6. In the second competition experiment, one third of the food suspension was renewed every 8 h, resulting in a much better approximation to a continuous resource supply. At 12 °C Synchaeta and Brachionus coexisted for more than 1 month and the densities of both rotifer species were significantly lower in the presence of their competitor. In contrast to expectations, Brachionus was able to persist even when Cryptomonas concentrations fell below its TFL. This was probably because Brachionus was using detritus and associated bacteria as additional food sources, which were present in the cultures during the later phase of the experiment. 7. Autocorrelation analysis of the temporal changes in egg ratios revealed significant periodic cycles in Synchaeta during the second competition experiment. A possible explanation for this is the fecundity schedule of Synchaeta, in which reproduction is highly concentrated in a few age classes. According to demographic theory, such a life cycle feature can cause slower convergence to a stable age distribution.     

Proline hydroxylation by cell free extract of a streptomycete

Free L-proline was hydroxylated to free L-hydroxyproline by cell free extract of Streptomyces griseoviridus P8648. The hydroxylation reaction required ferrous ion, 2-ketoglutarate and ascorbate. Zinc ion, ethylenediaminetetraacetic acid and alpha,alpha'-dipyridyl inhibited the reaction. Optimum temperature and pH were 25.0 degrees C and 7.5, respectively.     

Contrasting responses of photosynthesis at low temperatures in different annual legume species

Growth, net photosynthetic rate (P _N), chlorophyll fluorescence induction kinetics, and stromal fructose-1,6-bisphosphatase (sFBPase) in annual legumes native to the Mediterranean region, two clovers (Trifolium subterraneum L. ssp. oxaloides Nyman cv. Clare and T. michelianum Savi cv. Giorgia) and two Medicago species (M. polymorpha L. cv. Anglona and M. truncatula Gaertn. cv. Paraggio), shifted from 20 to 10 °C for 1 d or developed at 10 °C were compared with controls kept at 20 °C. Cold development produced a larger stimulation of growth in the clover cv. Giorgia and the Medicago cv. Paraggio. Transferring plants to low temperatures affected P _N relatively less in clovers than in Medicago plants. Development at 10 °C relieved the inhibition of photosynthesis in Giorgia and Paraggio, but not in Clare and Anglona, which correlated with increases in the maximum rate of carboxylation by ribulose-1,5-bisphosphate carboxylase/oxygenase, RuBPCO (V_cmax), and the photon-saturated rate of electron transport (J_max). In Medicago, transfer from high to low temperature inhibited photosynthesis in a lesser extent in Anglona than in Paraggio, which showed severe limitations at level of V_cmax and J_max. Development at 10 °C in Paraggio produced an efficient photosynthetic cold acclimation, this being associated with a two-fold increase of quantum yield of photosystem 2 electron transport (F/F'_m) and with the activity of sFBPase. By contrast, Anglona showed an irreversible inhibition of P _N coupled with the reduction of carbon metabolism by impairment of Calvin cycle enzyme activities such as RuBPCO and sFBPase, resulting in a poor cold acclimation of photosynthesis in this cultivar.     

Nature of collagens synthesized by monkey periodontal-ligament fibroblasts in vitro.

1. First subcultures of fibroblast-like cells from adult monkey periodontal ligament were incubated in the presence of 14C-labelled amino acids and produced significant amounts of type-I and type-III collagens. 2. The proportion of type-III collagen produced was calculated on the basis of the recovery of procollagens from DEAE-cellulose chromatography to be approx. 20%, and at least 10% when analysed as collagens on CM-cellulose chromatography. 3. Sodium dodecyl sulphate/polyacrylamide-gel electrophoresis of the procollagens, the collagens and their CNBr peptides was used to confirm the identity of the collagen types. 4. In serum-free media extensive conversion of type-I procollagen, but not of type-III procollagen, into collagen was observed, suggesting that a specific type-I procollagen peptidase was produced. 5. The pattern of collagen synthesis was not significantly different from that obtained with fibroblasts derived from skin corium of the same animals.     

A comparative study of the formation of extracellular proteins by Aeromonas salmonicida at two different temperatures

Aeromonas salmonicida was grown in a supplemented 3% (w/v) tryptone soya broth medium at 10°C, a temperature at the lower end of the range over which furunculosis has been observed to occur in the field, and 25°C, the optimum temperature for growth. Similar bacterial densities in the range 2.35 × 0.05 mg dry wt/ml were achieved in the two cultures at the beginning of the stationary phase of the growth cycle, after 125 h at 10°C and 18 h at 25°C. At this point, at the higher temperature 1.5 times more exoprotein was formed, 80 × 2.8 μg/ml compared with 54 × 1.7 μg/ml. Exoprotein contained the same proportion of haemolysin at both temperatures and twice as much protease at the higher temperature. The most marked difference was in an unidentified 100 kD protein which was formed in a 10-fold greater amount at 10°C.     

Developmental schedules and persistence of experimental host-parasitoid systems at two different temperatures

In experimental systems of a bruchid host, Callosobruchus chinensis, and a braconid parasitoid, Heterospilus prosopidis, the effects of changes in developmental schedules were examined in relation to the persistence of the system, or the time to extinction of a component species. We modified the developmental schedules by changing the temperature from 30°C to 32°C. To compare persistence, a long-term system with overlapping generations was set up and the bruchid host resource, azuki beans (Vigna angularis), were renewed every 10 days. The long-term systems showed greater persistence at 30°C than at 32°C. Parasitoid extinction was often observed. We examined differences in life-history characteristics of the component species between the two temperatures by short-term, single-generation experiments. Fecundity and egg hatchability of the host were reduced and the developmental period of the parasitoid was shortened at 32°C. The age at which the host became vulnerable to parasitoid attacks was earlier at 32°C than at 30°C. We constructed a daily based, age-structured model to analyse which life-history change(s) affected the persistence of the long-term systems. The density-dependent population growth of the host was described by a logistic equation and the attack rate of the parasitoid by a type II functional response with mutual interference. The simulation results showed greater persistence at 30°C than at 32°C. Sensitivity analysis showed that there are threshold boundaries in the length of the vulnerable period of the host beyond which system persistence drastically changes. Further, persistence at another temperature, 28°C, was predicted using a model based on short-term data on the host.     

A comparative study of the formation of extracellular proteins by Aeromonas salmonicida at two different temperatures

Aeromonas salmonicida was grown in a supplemented 3% (w/v) tryptone soya broth medium at 10 degrees C, a temperature at the lower end of the range over which furunculosis has been observed to occur in the field, and 25 degrees C, the optimum temperature for growth. Similar bacterial densities in the range 2.35 +/- 0.05 mg dry wt/ml were achieved in the two cultures at the beginning of the stationary phase of the growth cycle, after 125 h at 10 degrees C and 18 h at 25 degrees C. At this point, at the higher temperature 1.5 times more exoprotein was formed, 80 +/- 2.8 micrograms/ml compared with 54 +/- 1.7 micrograms/ml. Exoprotein contained the same proportion of haemolysin at both temperatures and twice as much protease at the higher temperature. The most marked difference was in an unidentified 100 kD protein which was formed in a 10-fold greater amount at 10 degrees C.     

Characterization of the protein species synthesized in vivo and in vitro by an aphid endosymbiont

The endosymbionts isolated from rifampicin-injected insects were less active in synthesizing proteins in vitro than were the uninjected ones. It was confirmed using [³H]leucine that the pea aphid, when injected with cycloheximide, actively synthesized only one protein species, symbionin. The synthesis of symbionin is highly sensitive to chloramphenicol. When separated two-dimensionally, symbionin was represented by a single spot. It is an acidic protein with mol. wt of about 63,000. Two days after an injection of cycloheximide the aphid began to synthesize a protein in addition to symbionin. The protein's mol. wt was less than that of symbionin and its synthesis was also sensitive to chloramphenicol. On a two-dimensional gel, proteins synthesized in the isolated endosymbiont were separated into more than 100 species. Under incomplete anaerobic conditions, the isolated symbiont synthesized several unique proteins.     

Calcium carbonate precipitation by two groups of moderately halophilic microorganisms at different temperatures and salt concentrations

Carbonate crystal formation by 48 strains of moderately halophilic microorganisms currently assigned to the genusFlavobacterium andAcinetobacter has been investigated. Strains were grown at different salt concentrations (2.5%, 7.5%, and 20%, wt/vol, total salts) and temperatures (22°C and 32°C). All the strains tested were capable of precipitating calcium carbonate as calcite, but onlyAcinetobacter strains formed aragonite. High temperature and low ionic strength of medium favored crystal formation. The influence of species specificity on the type of crystal precipitated by moderately halophilic microorganisms and their possible role in active precipitation in nature are discussed.     

Measured and predicted mineralization of clover green manure at low temperatures at different depths in two soils

Predictive models of the temporal mineralization pattern of organic residues may help in development of strategies to synchronize N mineralization with the crop demand and minimize off-season losses. In the present investigation, two double first-order models with temperature as a driving variable were tested against data on decomposition and N mineralization, respectively, in two field experiments with green manure. On 15 November 1984, mesh bags with red clover (Trifolium pratense L.) shoot material were placed at five depths (0–30 cm) on a sandy-loam and a loam site in south-eastern Norway. 167 days after burial, 73% of the initial clover nitrogen remained on the surface, 62% at 5-cm depth, and 56% at 30-cm depth. The differences among buried samples largely persisted throughout the experimental period (1.5 years). The decomposition rate slowed down appreciably after day 270, when the amount of N in buried bags averaged 33% of the initial N. The effect of site was small and varied during the experiment. The decay model, which was derived from laboratory incubations, predicted the initial observations of remaining clover material fairly well. Later, predicted and measured values diverged because recalcitrant residues decomposed more extensively in the field than in the laboratory. The N mineralization model was tested against net N mineralization from white-clover (T. repens L.) green manure ploughed down in late October. The course of the net N mineralization was well described when disregarding an over-prediction (6–12% of applied clover N), which may be due to N losses not accounted for in the model. The predictions were sensitive to the kind of function applied for correction of decay rates at temperatures below 0° C. The results showed that decomposition of clover green manure is rapid, even at temperatures below 5° C. N-rich plant material, therefore, should be worked into the soil as late as possible in the autumn or, preferably, remain on the soil surface until spring in order to reduce the probability of N losses.     

Effect of relaxin on the phenotype of collagens synthesized by cultured rabbit chondrocytes

The effect of porcine relaxin on rabbit articular and growth plate chondrocytes in primary culture was investigated by measurement of total collagen production and analysis of the phenotypes of newly synthesized collagen chains. A 24-h treatment of monolayer articular and multilayer growth plate chondrocytes with 2 micrograms per ml relaxin had no effect on total DNA and did not significantly modify the amount of [3H]proline-labelled collagen chains secreted by the cells. However, polyacrylamide gel electrophoresis demonstrated relevant modifications in relaxin treated chondrocytes. A significant increase was observed in the proportion of type III collagen and in the intensity of the band corresponding to alpha 2I chains. Two-dimensional peptide mapping of CNBr-cleaved molecules indicated that the band that was identified as alpha 1II on monodimensional gels contained a significant proportion of alpha 1I collagen chains, as demonstrated by the presence of alpha 1I cyanogen bromide-digested peptides. The intensity of this band was increased by relaxin treatment. Furthermore, total RNA analysis by slot blot and Northern blot techniques showed a dose-dependent stimulation of alpha 1I and alpha 1III mRNA levels after incubation with increased relaxin concentrations, but no change in the amount of alpha 1II mRNA. These results suggested that when added to cartilage cells in vitro, relaxin modulated the expression of type I, type II and type III collagen genes by amplifying the dedifferentiation process.     

Susceptibility to entomopathogens and modulation of basal immunity in two insect models at different temperatures

In this work, we analysed the efficacy of different commercial bio-insecticides (Steinernema feltiae, Steinernema carpocapsae, Heterorhabditis bacteriophora and Bacillus thuringiensis) by valuating the mortality induced on two insect models, Galleria mellonella (Lepidoptera) and Sarcophaga africa (Diptera) after exposure to different temperatures (10, 20 and 30 °C). Moreover, we investigated the effects of temperature on the basal humoral immunity of the two target insects; particularly, phenoloxidase (PO) and lysozyme activity. Our results show that G. mellonella is susceptible to all bio-insecticides at all the examined temperatures, except when infected at 10 °C with S. carpocapsae and at 30 °C with S. feltiae and B. thuringiensis. S. africa is more susceptible at 30 °C to all bioinsecticides; whereas, when infected at 10 and 20 °C, H. bacteriophora is the most efficient. Temperature modulates PO activity of both G. mellonella and S. africa, otherwise variations in lysozyme activity is observed only in G. mellonella. Except for a possible correlation between the increased lysozyme activity and the delayed Bt efficacy recorded on G. mellonella at 30 °C, a different resistance to bio-insecticides at different temperatures does not seem to be associated to variations of the host basal immunity, probably due to immunoevasive and immunodepressive strategies of these entomopathogens.