The effect of cold on platelets. II. Platelet function after short-term storage at cold temperatures.
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The heritability of packed red blood cell volume (PCV) and its correlation to gain in chicks brooded at a relatively cold (26.7 C) and a normal (32.2 C) environmental temperature were compared. The PCV of progeny from 28 sire families brooded at 26.7 C was 32.7% compared to 30.9% for progeny from the same sire families brooded at 32.2 C. The respective PCV heritabilities for the 26.7 C and 32.2 C groups were .51 and .57. There were no significant differences in gain for the groups brooded at different temperatures, and PCV was not correlated with gain.U
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The separate effects of ambient temperature and energy intake on the metabolism of thyroid hormones in the young pig have been investigated. Piglets were kept singly from 4 weeks of age at 10 or 35 degrees C on a high (H), or low (L) energy intake, such that H = 2L. At the tenth week of age blood samples were taken 16-21 h after the last meal for the determination of plasma concentrations of thyroxine (T4) and 3,5,3'-triiodothyronine (T3). At the end of the tenth week of age the fractional disappearance rate (K) of T4 was determined and finally the K value for T3 was estimated. Plasma concentrations of T4 and T3 were greater in animals on a high intake than in those on a low intake, (P less than 0.02), but the temperature at which the animals lived had no significant effect. The values of K for T4 and T3 were greater at 10 than at 35 degrees C (P less than 0.010 and P less than 0.05), and energy intake had no significant effect. Calculations of the catabolic rates in nmol/kg X h revealed that for T4 the rate was greater at 10 than at 35 degrees C (P less than 0.001) and on a high than on a low intake (P less than 0.01). Similar differences were found between the mean values of T3 with respect to both temperature and energy intake, but the variance was large and they did not reach statistical significance. It is concluded that the metabolism of thyroid hormones is influenced independently by both ambient temperature and energy intake.
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Lactobacillus sakei is a lactic acid bacterium widely represented in the natural flora of fresh meat. The aim of this study was to analyze the differences in protein expression during environmental changes encountered during technological processes in which L. sakei is involved in order to gain insight into the ability of this species to grow and survive in such environments. Using two-dimensional electrophoresis, we observed significant variation of a set of 21 proteins in cells grown at 4 degrees C or in the presence of 4% NaCl. Six proteins could be identified by determination of their N-terminal sequences, and the corresponding gene clusters were studied. Two proteins belong to carbon metabolic pathways, and four can be clustered as general stress proteins. A phenotype was observed at low temperature for five of the six mutants constructed for these genes. The survival of four mutants during stationary phase at 4 degrees C was affected, and surprisingly, one mutant showed enhanced survival during stationary phase at low temperatures.
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Shewanella sp. Ac10 is a psychrotrophic bacterium isolated from the Antarctica that actively grows at such low temperatures as 0 degrees C. Immunoblot analyses showed that a heat-shock protein DnaK is inducibly formed by the bacterium at 24 degrees C, which is much lower than the temperatures causing heat shock in mesophiles such as Escherichia coli. We found that the Shewanella DnaK (SheDnaK) shows much higher ATPase activity at low temperatures than the DnaK of E. coli (EcoDnaK): a characteristic of a cold-active enzyme. The recombinant SheDnaK gene supported neither the growth of a dnaK-null mutant of E. coli at 43 degrees C nor lambda phage propagation at an even lower temperature, 30 degrees C. However, the recombinant SheDnaK gene enabled the E. coli mutant to grow at 15 degrees C. This is the first report of a DnaK supporting the growth of a dnaK-null mutant at low temperatures.
PURPOSE: To examine the effects of a single 10-mg dose of ML on physical performance in EIB- and EIB+ athletes. METHODS: Twenty-four male college ice hockey players performed two 6-min maximal work accumulation bouts on an electronically braked cycle ergometer in subfreezing conditions (-2.5 +/- 0.4 degrees C) 6-8 h after either ML or placebo (PL) to obtain total work accumulated (kJ); subjects were evaluated for EIB after each exercise trial. RESULTS: Eight (33%) subjects were identified as EIB+ (23.5 +/- 13.35% fall in FEV1); 16 were EIB- (1.8 +/- 3.03% fall in FEV1). ML provided an approximately 50% protection against postexercise fall in FEV1. No significant differences in kJ were found between PL and ML trials for pooled subjects (95.3 +/- 13.69 and 94.8 +/- 13.27 kJ, respectively), EIB- subjects (99.6 +/- 13.26 and 99.0 +/- 11.81 kJ, respectively), or EIB+ subjects (86.8 +/- 10.67 and 86.5 +/- 12.72 kJ, respectively). Total work accumulated for EIB- subjects was significantly greater than for EIB+ subjects for both PL and ML (P < 0.05). CONCLUSION: A single 10-mg dose of ML had no ergogenic effect for EIB- and EIB+ subjects performing short-duration high-intensity exercise in subfreezing temperature, supporting the use of ML as EIB prophylaxis during international sport competition.
Hymenolepis diminuta grown in mice maintained at 5 degrees C were significantly larger and markedly more developed than those grown simultaneously in control mice maintained at 21 degrees C. In mice maintained at 5 degrees C, the incidence of infection and the number of worms recovered per host were higher than in the mice kept at 21 degrees C. Regardless of the temperature of the hosts' external environment, primary infections were always expelled before Day 13 postinfection and secondary (challenge) infections were invariably lost before Day 7.
The effect of low, nonfreezing temperatures on the viability of five strains of Chromobacterium violaceum was studied. The viability of cultures grown at 30 C was determined after exposure to various diluents held at 0 to 2 C. A culture diluted at its growth temperature served as the control. Cells of strain N were most sensitive in the early part of the exponential phase of growth. Cells of strains 252 and 341 were most sensitive in the late exponential, early stationary phase of growth. Cells of strain 9 showed greatest loss of viability during the maximal stationary phase. Strain 69 was completely resistant throughout its growth cycle to cold injury. Cell viability was much greater in buffered salts solution than in distilled water, broth, or physiological saline, whether cultures were diluted at room temperature or in the cold. The proportion of cells surviving after exposure to cold, however, was the same regardless of the composition of the diluent. Loss of viability was progressive at 0 to 2 C and reached a maximum after 2 hr. There was no loss of viability of cells exposed to 20 C, but there was some loss at 12 C. Strain 341 cultivated at 15 C was much less sensitive to 0 to 2 C than when it was cultivated at 30 C. The composition of the growth medium seemed to have no effect on the survival of cells exposed to cold. The polyamines, spermine and trimethylenediamine, as well as erythritol and sucrose, exerted some protective action against the effects of cold but not uniformly for all strains studied.
Descendants of two Colorado, U.S.A., populations of D. pseudoobscura, Boulder and La Foret, were exposed to +2 degrees and -2 degrees . In third instar larvae from Boulder AR and PP third chromosome gene arrangements survived better than TL and others, while the reverse situation occurred for La Foret. Deleterious dominant effects were observed for AR from La Foret. In adults from Boulder there was a trend towards greater survival for AR and PP than for other gene arrangements, while AR from La Foret showed relatively poor cold resistance. Survival of Boulder and La Foret flies, and their interpopulation hybrid, was determined after exposure to -2 degrees at two humidities. Order of survival of developmental stages was: adults >> third instar larvae > mixture of first and second instar larvae. Adults survived better at low humidity, while larvae survived better at high humidity. Boulder adults and larvae survived better than those from La Foret. Advantage in survival of hybrids over the midparent ranged from 23% to 138%. Hybrid advantage over the higher parent ranged from 5% to 111%. Order of expression of heterosis was: mixture of first and second instar larvae > third instar larvae > adults. Relation of all results to the chromosomal polymorphisms at Boulder (seasonally constant) and La Foret (seasonally cyclic) is discussed.
Refolding of GuHCl-denatured recombinant-human growth hormone (r-hGH) was investigated in both dilution additive and artificial chaperone assisted modes. In both techniques, it was found that CTAB is a better additive (in dilution mode) or a capturing agent (in artificial chaperone method). Neither of the two techniques was capable of complete inhibition of aggregates formed during refolding process. In dilution, using CTAB or alpha-cyclodextrin (alpha-CD) as two different additives, the aggregation was inhibited by almost 55%. However, the extent of inhibition raised to almost 82% in artificial chaperone assisted mode using CTAB as the capturing and alpha-CD as the stripping agents. Maximum inhibition of aggregation (up to 97%) was obtained when the entire process of refolding was done at 4 degrees C. However, under this temperature program, the far-UV CD and intrinsic fluorescence spectra of the refolded samples were not superimposable on their respective native spectra. The spectra superimposibilities were obtained when the refolding process was achieved under a well worked out temperature program: incubation of the sample for 3 min at 4 degrees C after initiation of the stripping step followed by overnight incubation at 22 degrees C. Based on these data, it is expected that higher activity recovery yields of recombinant proteins, particularly at relatively higher protein concentrations, could be achieved by getting a better molecular understanding of major factors responsive for aggregation and refolding pathways.
We tested the hypothesis that winter removal rates of fruits of wax myrtle, Myrica cerifera, are higher in colder winters. Over a 9-year period, we monitored M. cerifera fruit crops in 13 0.1-ha study plots in South Carolina, U.S.A. Peak ripeness occurred in November, whereas peak removal occurred in the coldest months, December and January. Mean time to fruit removal within study plots was positively correlated with mean winter temperatures, thereby supporting our hypothesis. This result, combined with the generally low availability of winter arthropods, suggests that fruit abundance may play a role in determining winter survivorship and distribution of permanent resident and short-distance migrant birds. From the plant's perspective, it demonstrates inter-annual variation in the temporal component of seed dispersal, with possible consequences for post-dispersal seed and seedling ecology.
Two strains of the spoiling bacterium S. putrefaciens showed an adaptation capacity to hyperosmotic shock when they were pretreated with a sublethal concentration of NaCl. The maximal tolerance factor for the CIP 69.29 strain was obtained when cells were incubated for 1 h in the presence of 1.5% NaCl, whereas for the J13.1 strain, an incubation of 15 min in the presence of 1% NaCl seemed to be the optimal conditions to harden the cells against a subsequent lethal salt treatment. During NaCl adaptation and growth at low temperatures (2 degrees C), 37 and 32 polypeptides were induced respectively. Interestingly, 11 proteins were common between the two different stress responses. These proteins and the corresponding genes seem to play a key role in the observed cross-protection towards the NaCl challenge induced by growth of the cultures at 2 degrees C. One of the overlapping proteins has been identified to correspond to the alkyl hydroperoxide reductase (AhpC) of S. putrefaciens. Northern blot analysis showed that induction of this enzyme was accompanied by accumulation of the corresponding transcript under both conditions.
The penetration of zona pellucida-free hamster ova by human spermatozoa has been used to quantitate sperm penetration potential. However, since mammalian eggs in vitro have limited viability, the effect of in vitro aging on the ability of hamster ova to be penetrated by human spermatozoa was examined. Zona-free ova maintained at room temperature (25 degrees C) lost their ability to be subsequently penetrated with a half-life of 50.1 +/- 8.8 minutes. This was partly the result of removing the zona pellucida by trypsin digestion, since zona-free oocytes in the presence of trypsin inhibitor or zona pellucida-intact oocytes had half-lives of 99.1 +/- 15.2 and 120.5 +/- 17.4 minutes, respectively. Reduction in penetration rates associated with ovum aging did not appear to be due to loss of viability and could be completely prevented by maintaining the ova on ice (4 degrees C). In the presence of TEST-yolk buffer at 4 degrees C, ova retained (100%) their ability to be penetrated for up to 24 hours and were morphologically indistinguishable from fresh ova. These observations show that ovum aging in vitro at 25 degrees C is much greater than previously anticipated. This may result in artifactually low and variable scores in the penetration bioassay.
Antioxidant activity, total phenolic content, anthocyanin content, and six other fruit characters including titratable acid concentration, soluble solids, firmness, and percentage of bruised berries were determined for nine blueberry (Vaccinium L. sp.) cultivars at harvest and at various postharvest intervals after storage at 5 degrees C. Berries from MSU-58, Brigitta, and Legacy stored successfully for 7 weeks, Bluegold stored for 3-5 weeks, Bluecrop, Elliott, and Nelson stored for 3 weeks, and Jersey and Little Giant stored for fewer than 3 weeks. During the time they retained marketable quality, one cultivar (MSU-58) demonstrated a 29% increase in antioxidant activity. None of the cultivars showed a significant decrease from the harvest antioxidant activity value during storage. Antioxidant activity, total phenolic content, and anthocyanin content were strongly correlated with each other (r = 0.87-0.99, P < 0.01). All three parameters were moderately correlated with soluble solids (r = 0.47, P < or =0.05; r = 0.44, P < or = 0.05; and r = 0.64, P < or = 0.01, respectively), and antioxidant activity and total phenolic content were both moderately correlated with pH (r = 0.53 and 0.49, respectively; P < or = 0.05). However, antioxidant activity, total phenolic content, and anthocyanin content showed no correlation with firmness, percent severely bruised berries, or weight loss. Antioxidant activity and total phenolic content at harvest both correlated with titratable acidity at harvest (r = 0.68, P < or = 0.05 and r = 0.70, P < or = 0.05, respectively) on a cultivar mean basis. Berries from Elliott were also harvested from plants at two levels of bush ripeness (30-50% and 60-80% ripe berries on plants) and separated into three fruit maturity classes on the basis of percent blue color. The level of bush ripeness had no significant effect on antioxidant activity, total phenolic content, or anthocyanin content; however, fruit maturity had a significant effect on antioxidant activity, total phenolic content, and anthocyanin content, and bush ripeness x fruit maturity interactions were significant for these three traits. Berries with 50-75% blue coloration harvested from bushes with 60-80% mature fruit showed a significant increase in antioxidant activity, total phenolic content, and anthocyanin content during the first 3 weeks in storage. Our results demonstrate that increases in antioxidant activity, total phenolic content, and anthocyanin content may occur in the blueberry during cold storage and are cultivar-dependent. The increases that occur in immature fruit, such as in Elliott, may be advantageous for producers who wish to delay marketing of the fruit.
Numerous lines of evidence indicate that the initial photoresponse of the circadian clock in Drosophila melanogaster is the light-induced degradation of TIMELESS (TIM). This posttranslational mechanism is in sharp contrast to the well-characterized pacemakers in mammals and Neurospora, where light evokes rapid changes in the transcriptional profiles of 1 or more clock genes. The authors show that light has novel effects on D. melanogaster circadian pacemakers, acutely stimulating the expression of tim at cold but not warm temperatures. This photoinduction occurs in flies defective for the classic visual phototransduction pathway or the circadian-relevant photoreceptor CRYPTOCHROME (CRY). Cold-specific stimulation of tim RNA abundance is regulated at the transcriptional level, and although numerous lines of evidence indicate that period (per) and tim expression are activated by the same mechanism, light has no measurable acute effect on per mRNA abundance. Moreover, light-induced increases in the levels of tim RNA are abolished or greatly reduced in the absence of functional CLOCK (CLK) or CYCLE (CYC) but not PER or TIM. These findings add to a growing number of examples where molecular and behavioral photoresponses in Drosophila are differentially influenced by "positive" (e.g., CLK and CYC) and "negative" (e.g., PER and TIM) core clock elements. The acute effects of light on tim expression are temporally gated, essentially restricted to the daily rising phase in tim mRNA levels. Because the start of the daily upswing in tim expression begins several hours after dawn in long photoperiods (day length), this gating mechanism likely ensures that sunrise does not prematurely stimulate tim expression during unseasonally cold spring/summer days. The results suggest that the photic stimulation of tim expression at low temperatures is part of a seasonal adaptive response that helps advance the phase of the clock on cold days, enabling flies to exhibit preferential daytime activity despite the (usually) earlier onset of dusk. Taken together with prior findings, the ability of temperature and photoperiod to adjust trajectories in the rising phases of 1 or more clock RNAs constitutes a major mechanism contributing to seasonal adaptation of clock function.