Injury to potato leaves exposed to subzero temperatures in the absence of freezing

Electrolyte leakage was measured in hardened and nonhardened leaves of three potato species, Solanum tuberosum L., S. acaule Bitt. and S. commersonii Dun., and one interspecific cross, Alaska Frostless (S. acaule x S. tuberosum) when exposed to various subzero temperatures. The leaves were undercooled (no ice present) from 0°C to -12.5°C for 45 min and to-4°C for up to 10 d. Regardless of the degree of undercooling no injury was observed in any of the potatoes, hardened or nonhardened, for up to 12 h. After 5 d, however, electrolyte leakage was observed in hardened S. tuberosum, S. acaule and S. commersonii, and in nonhardened Alaska Frostless. After 10 d exposure all potatoes, hardened and nonhardened, showed a significant amount of electrolyte leakage as compared to their controls kept at 0°C for 10 d.Scientific Journal Series Paper No. 13842 of the Minnesota Agricultural Experiment Station, St. Paul, Minn     

Different tolerance to hypothermia and rewarming of isolated rat and guinea pig hearts.

We examined the effect of hypothermia and rewarming on myocardial function and calcium control in Langendorff-perfused hearts from rat and guinea pig. Both rat and guinea pig hearts demonstrated a rise in myocardial calcium ([Ca]total) in response to hypothermic perfusion (40 min, 10 degrees C), which was accompanied by an increase in left ventricular end diastolic pressure (LVEDP). The elevation in [Ca]total was severalfold higher in guinea pig than in rat hearts, reaching 12.9 +/- 0.8 and 3.1 +/- 0.6 micromol.g dry wt-1, respectively. The rise in LVEDP, however, was comparable in the two species: 62.5 +/- 2.5 (guinea pig) and 52.5 +/- 5.1 mm Hg (rat). Following rewarming, [Ca]total remained elevated in guinea pig, whereas a moderate decline in [Ca]total was observed in the rat (13.6 +/- 1.9 and 2.2 +/- 0.3 micromol.g dry wt-1, respectively). Posthypothermic values of LVEDP were also significantly higher in guinea pig compared to rat hearts (42.5 +/- 6.8 vs 20.5 +/- 5.1 mm Hg, P < 0.027). Furthermore, whereas rat hearts demonstrated a 78 +/- 7% recovery of left ventricular developed pressure, there was only a 15 +/- 7% recovery in guinea pig hearts. Measurements of tissue levels of high energy phosphates and glycogen utilization indicated a higher metabolic requirement in guinea pig than in rat hearts in order to oppose the hypothermia-induced calcium load. Thus, we conclude that isolated guinea pig hearts are more sensitive to a hypothermic insult than rat hearts.     

Growth, photosynthesis and assimilate partitioning in Lolium temulentum exposed to chilling temperatures

Vegetative plants of Lolium temulentum L, grown at 20°C with an 8 h photoperiod were transferred to either 5 or 2°C (8 h photoperiod) at 4th leaf maturity. Measurement of dry weight gain indicated a marked decline in relative growth rate below 5°C, with growth being reduced as much between 5 and 2°C as between 20 and 5°C. This reduction was not associated with increased mortality and was reversible if plants were returned to 20°C. Tissue explants from cold-treated plants retained the ability to extend if returned to 20°C Rates of extension in explants were less temperature sensitive than the relative growth rates observed in intact plants. Measurements of photosynthetic capacity, and of the patterns of accumulation of reserve carbohydrate in plants exposed to different temperatures, suggested that the inhibition of growth caused by chilling is not caused by an inability of the plants to fix sufficient carbon.     

Sensitivity of pigmented mucosa and skin to freezing injury.

Pigmented areas of canine skin and oral mucosa were subjected to freezing in situ at various temperatures for the purpose of investigating possible differences in the sensitivity of epidermal cells to cold injury. Destruction of melanocytes occurred in the range of ?4 to ?7 °C, while squamous cells resisted injury even at ?20 °C. Replacement of the lost pigmented cells occurred from the normal tissue at the periphery of the injured area. The experiments suggest that selective destruction of pigmented lesions in clinical conditions may be achieved by freezing tissue to ?4 to ?7 °C or colder, but not to exceed ?20 °C in order to avoid destruction of squamous cells. The experiments also support wider use of cryosurgery for pigmented lesions of the skin and oral cavity.     

Vitrification of goat, sheep, and cattle skin samples from whole ear extirpated after death and maintained at different storage times and temperatures

Proper tissue preservation from a wide range of animals of different species is of paramount importance, as these tissue samples could be used to reintroduce lost genes back into the breeding pool by somatic cloning. We aim to study the temporal and thermal post-mortem limits, tested in rabbits and pigs, within which there will be guarantees of obtaining living skin cells in goat, sheep, and cattle. We also intend to study the effect of vitrification on the ability of ear skin cells, stored at different times and temperatures, to attach to the substratum and grow in vitro after warming. Ears were stored either at 4 degrees C for 12, 252, and 348 h post-mortem (hpm), or at room temperature (22-25 degrees C) for 60 and 96 hpm. In all cases, skin samples from these ears were sorted into two groups: one group was in vitro cultured immediately after storage, and the other group was vitrified after storage and further in vitro cultured. In goat and sheep, no differences in attachment (100%: goat; 90-100%: sheep) or subconfluence (75-100%: goat; 70-100%: sheep) rates were observed between experimental groups. However, in days of culture to reach subconfluence, significant differences between non-vitrified and vitrified groups were observed when ears were stored at 4 degrees C for 12 and 252 hpm. In cattle, with respect to attachment rate, vitrified samples from ears stored at 22-25 degrees C for 60 hpm were different from non-vitrified control group (60 vs. 100%, respectively; P < 0.05). Also, days of culture to reach subconfluence were analysed by a non-parametric Cox Survival Analysis. In general, results from ANOVA and Survival Analysis were similar, because the proportion of censored data was quite low (9%), so the bias when using ANOVA is not too high. In spite of all the above, the lowest survival rates (75%: goat; 70%: sheep; and 40%: cattle) were sufficiently high to enable collection of skin samples from the majority of dead animals and their cryopreservation.     

Nitrate reductase activity, nitrogenase activity and photosynthesis of black alder exposed to chilling temperatures

Actinorhizal ( Frankia -nodulated) black alder [ Alnus glutinosa (L.) Gaertn.] seedlings fertilized with 0.36 m M nitrate (low nitrate fertilizer treatment) or 7.14 m M nitrate (high nitrate fertilizer treatment) and acclimated in a growth chamber for 2 weeks were exposed to 2.5 h of night-time chilling temperatures of −1 to 4°C. Cold treatment decreased nitrogenase activity (acetylene reduction activity) 33% for low nitrate fertilized plants and 41% for high nitrate fertilized plants. Recovery of nitrogenase activity occurred within 7 days after chilling treatment. In contrast, in vivo nitrate reductase (NR) activities of leaves and fine roots increased immediately after chilling then decreased as nitrogenase activities recovered. Fine roots of alder seedlings exhibited NR activities proportional to the amounts of nitrate in the rooting medium. In contrast, the NR activities of leaves were independent of substrate and tissue nitrate levels and corresponded to nitrogenase activity in the root nodules. In a separate experiment, net photosynthesis (PS) of similarly treated black alder seedlings was measured before and after chilling treatments. Net PS declined in response to chilling by 17% for plants receiving low nitrate fertilizer and 19% for plants receiving high nitrate fertilizer. After chilling, stomatal conductance (g_s) decreased by 39% and internal CO₂ concentration (c_i) decreased by 5% in plants receiving the high nitrate fertilizer, whereas plants receiving the low nitrate fertilizer showed no change in g_s and a 13% increase in c_i. Results indicate that chilling stimulates stomatal closure only at the high nitrate level and that interference with biochemical functions is probably the major impact of chilling on PS.     

Thermoregulatory responses of Japanese Quail to various air temperatures and humidities

Measurements of rectal temperature (T_re), feather surface temperature (T_s), heart rate (HR) and breathing rate (BR) of adult Japanese Quail were taken at combinations of four mean relative humidities (rh) and stable or rising air temperature (T_a). T_re increased about 1.7° and 2.5°C at a stable T_a of 40°C from a T_a of 25°C at low (<50%) and high (> 80%) rh, respectively. T_s passively increased with T_a while remaining above T_a as high as 41°C. Panting and gular flutter rates were synchronous and approached rates of 800/min at high rh and stable T_a of 40°C. The steady-state panting rate was a direct curvilinear function of ambient water-vapor pressure (P_w). HR was depressed at successively higher, stable T_a's or with slowly increasing T_a. This thermal bradycardia may be related to the degree of discomfort or stress experienced by the animal.

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Zusammenfassung Bei Kombinationen von 4 relativen Feuchtigkeiten (rh) und konstanter oder ansteigender Temperatur (T_a) wurden Messungen der Rektaltemperatur (T_re), Federoberflächentemperatur (T_s), Herzfrequenz (HR) und Atemfrequenz (BR) an ausgewachsenen japanischen Wachteln vorgenommen. Bei T_a 40°C stieg T_re zwischen 1,7°–2,5°C an ausgehend von T_a 25°C bei <50% und > 80% rh. Die T_s folgte diesem Anstieg von T_a und überschritt ihn bis auf 41°C. Hecheln und Gurren stiegen synchron an bis auf 800/min bei hoher rh und 40°C T_a. Die erhöhte Hechelfrequenz war eine direkte lineare Funktion des Wasserdampfdruckes. HR war verlangsamt bei erhöhter, konstanter T_a. Die Hitzebradykardie steht vermutlich in enger Beziehung zu dem Ausmass der von dem Tier empfundenen Belastung.

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Resume On a soumis des cailles du Japon adultes à 4 taux différents d'humidité relative (rh) par température constante ou en hausse et mesuré la température rectale (T_re), la température superficielle des plumes (T_s), les battements du coeur (HR) et la vitesse de respiration (BR). T_re a augmenté de 1,7°C, respectivement de 2,5°C par suite d'une hausse de T_a de 25°C à 40°C par humidité basse (<50%), respectivement élevée (>80%). T_s a suivi la hausse de T_a et l'a même dépassée jusqu'à atteindre 41°C. Halètement et palpitations ont suivi la courbe de T_a pour atteindre 800/min par rh élevée et 40°C de T_a. La vitesse de respiration s'est élevée en fonction linéaire de la pression de vapeur. HR se sont ralentis par T_a élevée, mais constante. La bradycardie de chaud est probablement étroitement liée à la contrainte ressentie par l'animal.

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This investigation was supported in part by funds provided for medical and biological research by State of Washington Initiative Measure No. 171.     

Microbiological deterioration of Gambara's frescos exposed to open air in Brescia,Italy

Microbiological studies were carried out on samples taken from L. Gambara's deteriorated frescos, painted on outside walls of two buildings in Brescia, Italy, in the 16th century. The results showed that aerobic heterotrophic bacteria, fungi, ammonifiers and sulphur-oxidizing bacteria were present in all samples and in most cases were well above the levels considered damaging; only fungi and sulphur-oxidizing bacteria counts underwent variations relating to climatic conditions. Moreover, some fungi are able to grow in the presence of malachite and haematite, used as pigments for frescos, but not limonite. The same fungi change the green colour of malachite into blue.     

Differentiation of freezing tolerance and vernalization responses in Cruciferae exposed to a low temperature

High levels of freezing tolerance were induced in leaves of different Cruciferae species including Brassica napus, Arabidopsis thaliana, Barbarea vulgaris, Thlaspi arvenses and Descurainia sophia by low-temperature acclimation. Concomitantly, the amount of total RNA doubled in these three species. Analyses of methylation patterns and dosage of rRNA genes were carried out to determine whether or not alterations occur in this DNA during development of freezing tolerance. Hybridizations of Southern transfers with an rDNA probe revealed two additional EcoRI sites in purified DNA isolated from freezing-tolerant leaves of winter B. napus cv Jet Neuf and D. sophia (both of which require low temperatures for vernalization), but not in isolates of A. thaliana or spring B. napus cv Topas. An increase of rDNA cistrons was also observed in both B. napus cv Jet Neuf and D. sophia but not in A. thaliana or B. napus cv Topas upon cold acclimation. These results suggest that low temperature induced amplification of rDNA and the differential methylation of EcoRI sites may possibly be related to the vernalization process but may not be related to the development of freezing tolerance. However, the higher activity of RNA polymerase (2.5 times more) observed upon cold acclimation may explain the concomitant increase in total RNA and may be related to the development of freezing tolerance in the Cruciferae.     

Serum leptin, energy budget, and thermogenesis in striped hamsters exposed to consecutive decreases in ambient temperatures

Leptin has been found to be a direct participant in the regulation of both energy intake and energy expenditure in small mammals showing seasonal declines in body mass (M(b)) and fat mass, but its roles in an animal exhibiting seasonally increased thermogenesis and unchanged M(b) remain unclear. Serum leptin levels, energy budget, and thermogenesis were measured in striped hamsters exposed to consecutive decreases in ambient temperatures ranging from 23° to -23°C. Cold-exposed hamsters had significant increases in gross energy intake (GEI), the rate of basal metabolism, nonshivering thermogenesis, and activity of cytochrome c oxidase (COX) in brown adipose tissue (BAT), compared with control hamsters, indicating a cold-induced elevation of thermogenesis. Body mass and fat content were decreased in cold-exposed animals, and serum leptin levels were increased in hamsters exposed to temperatures of -8°C and below in inverse proportion to body fat content. Serum leptin levels were positively correlated with GEI and BAT COX activity in cold-exposed hamsters, but no such relationships were observed in control animals. These findings suggest that cold-exposed hamsters increase food consumption to meet the energy requirements for increased BAT thermogenesis. The increases in serum leptin levels are likely involved in increased thermogenesis in hamsters under cold stress. Cold-exposed hamsters may become leptin resistant, which is associated with impaired regulation of food intake. This new natural model of leptin resistance may also provide insight into the dynamic long-term control of energy homeostasis for animals that do not exhibit seasonal decline in M(b).     

Glutathion content, rate of apoptosis, and brown adipose tissue mass in rats exposed to different ambient temperatures

1. Amount of glutathione (GSH) in the interscapular brown adipose tissue (IBAT) of cold-adapted (5±1 °C), and 1, 3 and 7 days re-adapted (22±1 °C) rats were examined.

2. The higher amount of GSH was found in the IBAT of cold-adapted rats compared to control animals (22±1 °C).

3. After 1 and 7 days of re-adaptation, the level of GSH was still significantly higher compared to control group.

4. On the third day of re-adaptation, the quantity of GSH in the tissue was significantly lower compared to the cold-adapted group.

5. In cold-adapted animals, intensity of apoptosis in IBAT was lower, whereas in re-adapted animals (3rd and 7th day) it was higher than in control group.

6. This study indicates that GSH depletion is associated with an enhanced apoptosis in IBAT during re-adaptation.

Susceptibility of Tribolium castaneum life stages exposed to elevated temperatures during heat treatments of a pilot flour mill: influence of sanitation, temperatures attained among mills floors, and costs

The influence of sanitation on responses of life stages of the red flour beetle, Tribolium castaneum (Herbst) (Coleoptera: Tenebrionidae), was investigated in a pilot flour mill subjected to three, 24-h heat treatments by using forced-air gas heaters fueled by propane. Two sanitation levels, dusting of wheat flour and 2-cm-deep flour, were created in 25 plastic bioassay boxes, each holding eggs, young larvae, old larvae, pupae, and adults of T. castaneum plus two temperature sensors. Data loggers (48) were placed on the five mill floors to record air temperatures. The time required to reach 50 degrees C, time above 50 degrees C, and the maximum temperature among mill floors and in bioassay boxes were measured. The maximum temperature in bioassay boxes and in the mill was lower on the first floor than on other floors. This trend was apparent in time required to reach 50 degrees C and time above 50 degrees C, especially in compartments with 2-cm-deep flour. The mean +/- SE mortality of T. castaneum life stages on the first floor was 55.5 +/- 12.9-98.6 +/- 0.8%; it was 93.2 +/- 6.7-100 +/- 0.0% on other floors. Adults were the least susceptible stage. Mortality of T. castaneum stages in compartments with 2-cm-deep flour was generally lower than those with flour dust. Costs for the three heat treatments ranged from US$27,438 to $28,838. An effective heat treatment can be conducted within 24 h, provided temperatures on mill floors reach 50 degrees C in 8-12 h and are held above 50 degrees C for at least 10-14 h, with maximum temperatures held between 50 and 60 degrees C.     

Ecophysiological behavior of Caquetaia kraussii (Pisces: Cichlidae) exposed to different temperatures and salinities.

Tropical river sardine, Caquetaia kraussii, captured from La Aguá lagoon (Sucre State, Venezuela) were acclimatized for four weeks at 22, 24, 30 and 32 degrees C and at 0, 5, 10, 15 and 17@1000 salinity. To evaluate effects of thermal response to acclimatization level, the fish were transferred suddenly from lower temperatures (22 and 24 degrees C) to higher ones (32 and 30 degrees C) respectively. Then thermal resistance time was measured at the lethal temperature of 40.9 degrees C for 30 days. We considered that acclimatization process completed when resistance time was stabilized at the new temperature regime. For the saline effect, the concentrations of sodium and potassium were measured in the tissues at each treatment: gills, white muscle, gut and heart. The results showed that thermal tolerance increased rapidly in 3 h with a 6 degrees C rise in temperature (from 24 to 30 degrees C) and in 24 h with a 10 degrees C rise (22 to 32 degrees C). With decreasing temperatures, the acclimatization level reached its lowest in 11 days with a 6 degrees C decreases (from 30 to 24 degrees C) and in 14 days with a 10 degrees C decrease (32 to 22 degrees C). Caquetaia kraussii regulates as much sodium as potassium in gills and white muscle tissues at all salinity levels tested; however, gut and heart tissues showed significantly different regulations among salinities examined.