Effects of hyperthermia on enzymes and electrolytes in blood and cerebrospinal fluid in dogs

The effects of exposure to various degrees of heat stress on serum glutamate—oxaloacetate transaminase (SGOT), serum glutamate-pyruvate transaminase (SGPT), alkaline phosphatase (ALK-P-ase), calcium and chlorides have been studied on 75 dogs. Rectal temperature (T_re) was recorded before and after exposure to heat stress. These dogs were divided into 5 groups, according to the T_re level attained after exposure to heat stress. Rectal temperature was raised from normal to 39.45±0.47C in the first group, to 40.93±0.17C in the second group, to 41.87±0.22C in the third group, to 42.90 ± 0.21C in the fourth group and to 43.93±0.19C in the fifth group. The concentration of enzymes SGOT, SGPT and ALK-P-ase in blood and cerebrospinal fluid (CSF) increased significantly with hyperthermia. Calcium and chlorides concentrations in blood and in CSF tended to increase in hyperthermia. The integrity of the blood brain barrier for these enzymes and calcium is maintained under mild hyperthermia but it breaks down partially under influence of more severe hyperthermia. Core temperature above 41C results in damage to tissues and consequential rise of plasma enzymes. This degree of hyperthermia also seems to mark the beginning of injury to blood brain barrier. Critical core temperature tolerated by 50% of animals was 44C.     

Interaction of cytochrome c L with methanol dehydrogenase from Methylophaga marina 42:

The reaction of methanol dehydrogenase with cytochrome c _L from Methylophaga marina and the reactions of the non-physiological substrates, Wurster's blue and ascorbic acid, with both proteins were studied as a function of temperature (4–32 °C), pressure (1–2000 bar) and ionic strength using the optical high pressure stopped-flow method. The thermodynamic parameters H, S and V were determined for all reactions where electron transfers are involved. These data allowed the determination of the Maxwell relationships which proved the internal thermodynamic consistency of the system under study. A conformational change on the cytochrome c _L level was deduced from both breaks in the Arrhenius plots and the variation of the V with temperature.Abbreviations MOPS 4-morpholinepropanesulfonic acid - CHES 2-(cyclohexylamino)ethanesulfonic acid - MDH methanol dehydrogenase - EDTA ethylenedinitrilotetraacetic acid disodium salt - BTB bromothymol blue (3,3-dibromothymolsulfoneph-thalein) - PQQ 2,7,9-tricarboxy-lH-pyrrolo-[2,3f]quinoline-4,5-dione - cytochrome c _HH mammalian horse heart cytochrome c     

Central nervous versus total body thermosensitivity of the duck

Ducks were chronically implanted with thermodes in the POAH region, the lower brainstem or the vertebral canal. At thermoneutral conditions, lowering the temperature of the spinal cord (T_vc) or the lower brainstem (T_mb) stimulated metabolic heat production (M) with a subsequent rise of core temperature (T_c). Lowering the temperature of the POAH region (T_hy) induced a fall of T_c due to paradoxical activation of heat defence and, thus, induced slight to moderate general hypothermia depending on the cooling intensity. When T_hy was normalized, the hypothermia temporarily stimulated metabolic heat production until T_c was normalized. Cold sensitivity of the entire body, as revealed by the metabolic response to the hypothermia induced by preceding POAH cooling, and cold sensitivity of the spinal cord and the lower brainstem, as revealed by the metabolic response to local cooling, were quantified by calculating the quotient M/T from the maximum metabolic response and the experimentally induced drop of T_c, T_mb and T_vc. With lower brainstem cooling M/T_mbdid not exceed –0.4 W/(kg · C). With spinal cord cooling, M/T_vc did not exceed –0.6 W/(kg · C). The mean value of M/T_c after hypothermia induced by POAH cooling was –4.02 W/(kg · C). The results indicate that the cold sensitivity residing in the CNS of ducks represents only a small fraction of the entire cold sensitivity of the body.Presented at the Eighth International Congress of Biometeorology, 9–14 September 1979, Shefayim, Israel.     

Thermoregulation and thermal perception in the cold and heat before and after intermittent heat adaptation

Students wearing swim suits were exposed for 30 min to neutral room temperature (T_R=28C). During the following 60 min they were subjected to gradual decreases or increases of room temperature reaching 12C or 45C, respectively. Static thermal stimuli were applied to the palms of the right (38C) and left (25C) hands. Hands and feet of all subjects were thermally isolated at 22C ambient temperature. General thermal comfort (GTC), local thermal comfort (LTC), skin blood flow (which is proportional to heat transport index ) several body temperatures, oxygen-consumption , and sweat rate (S), were measured. After moderate intermittent heat exposures (7 times for 1h at T_R=42.5C) the experiments started again. From GTC, LTC, or as functions of T_R, no new knowledge about thermoregulatory or adaptive mechanisms was available. The high in the cold stimulated left hand, however, and the oscillatory thresholds (_OSC) for rhythmic vasomotion indicated the peripheral influence of skin temperature, as well as local, mean skin temperature (¯T_s) and core temperature. When exposed to moderate temperature decreases or increases the body seems to react only with increasing thermal resistance by vasoconstriction or an increase of sweat rate, respectively. Moderate heat adaptation is only able to raise sweat rate, but not the thresholds and gain of the S-function. We assume that functional studies of adaptive modifications in humans must be conducted at temperatures greatly beyond those used in these experiments.     

Variations in mental performance under moderate cold stress

Effects of moderate cold stress on reasoning ability, associative learning and critical flicker frequncy of Indian subjects were studied by exposing them to 25C,. 20C, 15C and 10C for three hours. A second set of experiments was also conducted to confirm the conclusions of the first by using the same temperatures and duration of exposure. However, not only the sample used in the second case was larger and different but also the mental functions tested were numerical ability, running memory and mental alertness. It has been concluded that there is a significant impairment of simple cognitive functions at 15C which is 10C lower than their most comfortable temperature of 25C.     

Growth temperature acclimation byAmoeba proteus: Effects on cytoplasmic organelle morphology

Summary The effects on subcellular morphology of maintaining amoebae at temperatures other than 20 C (the routine culture temperature) were assessed. Estimations of cycling potential at each temperature confirmed that acclimation had affected gross cell functioning. Generation times ranged from no division at 6 C, to an optimal minimum of 2 days at 22 C.Organelle morphology changes were studied after 5 days of growth at the new temperatures; alterations were most evident at the extremes of 6 and 28 C. The main mitochondrial alteration resulted in changes to the ratio of Type I: Type II organelles; with a decrease in Type I forms away from the optimal range of 20–22 C. Extended culturing at 6 C generated mitochondrial matrical inclusions. Ribosomal attachment to the endoplasmic reticulum, a common feature of 20 C-grown cells, decreased at the temperature extremes, where an increase in free ribosomes occurred. Upon extended culture at 6 C helical structures, usually observed in groups only within the nucleus, were also present in the cytoplasm. Golgi complexes were less common in cells maintained at extreme temperatures and often showed differences in shape. These changes were all reversible on a return to culturing at 20 C.The possible functional significance of these findings is discussed.     

Responses in rectal and skin temperatures to centrifugal forces in rats of different ambient temperatures

Effects of centrifugation upon rectal (T_re) and tail skin temperatures (T_s) were studied in male Wistar rats at varying ambient temperature (T_a) using a centrifuge which was placed in a climatic chamber. Centrifugal forces of Gz of 3.0 were imposed on rats which were suspended at horizontal body position using a newly developed mesh suits holding method in the animal box placed on the rotating arm of the centrifuge. Headwards or tailwards forces were applied according to the experimental design. No significant difference of the responses was observed between the two force directions.Centrifugations imposed at different T_a of 15, 20, 25, 30 and 32.5C resulted in falls in T_re accompanied by rises in tail T_s at the cooler environments, while rises in T_re accompanied by falls in T_s in the warmer environments. The T_a at which the response pattern of T_re and T_s was reversed (critical ambient temperature) was 26.8±2.3 (mean and SE) and 27.9±2.8C, respectively. Tolerance to centrifugation was markedly increased in cooler environments than in wanner ones. It was suggested that the increased skin pressure due to centrifugation exerted some inhibitory effects upon central thermoregulatory ability.     

Rhodopsin phosphorylation occurs at metarhodopsin II level

Photolyzed rhodopsin was phosphorylated in bovine rod outer segments incubated at –10 C. In the experiment in which urea-treated outer segments and rhodopsin kinase were incubated with ATP in the presence of 30% glycerol, the extent of phosphate incorporation at –10 C was about 30% of that at 37 C. Separation of phosphorylated rhodopsin by isoelectric focusing indicated that a limited number of sites were phosphorylated at –10 C. The partially phosphorylated pigment incorporated more phosphates when the temperatures was raised to 37 C. This was partly due to decreased inhibition of phosphorylation by glycerol at higher temperature. Since the maximum phosphorylation at –10 C (at which metarhodopsin II is stable) occurred at a pH value (6.0) lower than the pKa for metarhodopsin I-metarhodopsin II transition, metarhodopsin II was suggested to be the preferred substrate for rhodopsin kinase at –10 C. Limited proteolysis with thermolysin of rhodopsin phosphorylated at 37 C released peptides containing about 50% of the total phosphate incorporated. In contrast, proteolytic digestion of rhodopsin phosphorylated at –10 C released negligible amounts of phosphate-containing peptides. The results were taken to suggest that the incorporation of phosphates at metarhodopsin II level under the present condition occurred in the residues other than those removed by thermolysin digestion.Based on material presented at the Fifth International Congress of Eye Research, Eindhoven, October 1982     

Modulation of natural killer activity by thymosin alpha 1 and interferon

Summary A single injection of -interferon (-IFN) (30 000 units/mouse), a major biological modifier of natural killer (NK) cytolytic activity, strongly stimulated NK activity in normal mice, as expected, while the same treatment did not statistically alter the NK response in cyclophosphamide (CY)-suppressed animals.We investigated the possibility of thymosin ₁ cooperating with -IFN in boosting NK activity in CY-suppressed animals.The results show that treatment with thymosin ₁ (200 g/kg) for 4 days, followed by a single injection of -IFN 24 h before testing, strongly restored NK activity in CY-suppressed mice. Thymosin ₁ was, moreover, able to accelerate the recovery rate of NK activity in bone marrow reconstituted murine chimeras.Taken together the data support the concept that the synergic effect between thymosin ₁ and -IFN could be the result of effects on differentiation of the NK lineage at different levels.     

Effect of protons and cations on chloroplast membranes as visualized by the bound ANS fluorescence

Structural changes in the chloroplast membranes caused by acidification and heat-treatment are studied by observing the changes in the fluorescence of ANS bound to thylakoid membranes. On addition of acids to buffered suspension of isolated pea chloroplasts, the fluorescence intensity of bound ANS shows a sigmoidal rise on reaching a pH value of about 4.5. A part of the fluorescence enhancement of bound ANS brought about by protons is not reversible on back titration with alkali. The reversible part of acid induced rise in ANS fluorescence possibly reflects structural changes expected to be associated with photophosphorylation. Divalent cations enhance the fluorescence of ANS bound to chloroplasts between a pH range 4.5–7.0 but diminish it if the pH is below 4.5.Addition of acid to heat-treated chloroplasts shows similar sigmoidal rise in ANS fluorescence intensity on lowering the pH to about 4.5. On addition of acid upto a pH of 3.1, the ANS fluorescence is greater than that of untreated chloroplasts, however, at pH below 3.1, the fluorescence of bound ANS is lower than the control chloroplasts. This observation indicates that heat-treatment caused some alteration of the microstructure of thylakoid membranes of chloroplasts besides the usual loss in the O₂ evolving capacity.This is further confirmed from the studies of Hill-activity and ANS binding to chloroplasts incubated at various temperatures in the absence and presence of aliphatic alcohol. Hill-activity (DCPIP reduction) of chloroplasts incubated at temperatures between 25 C and 55 C first increases reaching a maximum at 45 C and then declines rather sharply, when the chloroplasts are heated beyond 45 C (Tmax). The presence of 200 mM n-butyl alcohol or 40 mM n-amyl alcohol during the warming treatment lowers the temperature by 8 C at which the decline in the Hill-activity is observed. An enhancement in the fluorescence intensity and a blue shift of the emission spectrum of bound ANS are noted if the chloroplasts are heated beyond the Tmax either in absence or presence of alcohol. The changes in the fluorescence of ANS bound to heat-treated chloroplasts plausibly reflect the nature of the structural changes in chloroplasts during the heating upto 55 C.Abbreviations ANS 1-anilino-8-naphthalene sulphonate - DCPIP 2,6-dichlorophenol indophenol     

Effects of climate on the thermoregulatory responses of male buffalo (Bubalus-bubalis) calves supplemented with different levels of testosterone and TDN

The present work was undertaken to study the changes in the thermoregulatory responses due to changing climate, androgen and TDN percent in 12 buffalo male calves from 4 months of age for a period of two years. The results obtained were: (1) The amplitudes of the diurnal cycles in rectal temperature (T_re) in each month of the experiment were very similar but longer in summer months as compared to winter months, (2) The correlation between T_re and ambient temperature (T_a) was high in all months. (3) Buffalo calves without testosterone supplementation on 70% TDN showed maximum rise in T_re by 0.076 0C per degree of ambient temperature, indicating more strain, (4) Highest respiratory frequencies occurred at 10:00 h during summer months. Subgroup 2 of group 1 showed the maximum rise by 311% in respiratory frequency in summer over winter due to least acclimatization, (5) The mean daily increase by 9.94C in ambient temperature doubled the respiratory frequency of male buffalo calves from January to April, and (6) Androgen therapy and higher TDN percent level helped the male buffalo calves to adjust better and sooner than the other calves to changing climates.     

The relative influence of urban climates on outdoor human energy budgets and skin temperature II. Man in an urban environment

Four typical urban neighborhoods or street canyon settings (including street parks) were simulated. These urban morphologies were exposed to typical summer and winter climatic scenarios for latitudes 10, 34, and 50N. The changes induced in the components of the human energy budget were examined. Resultant skin temperatures were compared with non-urban, unobstructed environments.     

Temperature dependent structural changes of intraphage T7 DNA

The phenomena connected with the first phase transition step of the native T7 phage at 40C–65 C have been studied using various methods. In this temperature range a) the optical melting curve shows an absorption decrease, b) the maximum of the small-angle X-ray scattering characteristic for DNA packing disappears, c) there is a drop of biological activity and d) there are changes in the structure of the difference absorption spectra of native phages versus isolated DNA. All data are interpreted assuming a structural change of the DNA due to the release of its protein coat towards the end of the first phase transition step (at 60–65 C in the case of M9 buffer). Above this temperature the intraphage DNA packing appears to be destroyed and the DNA structure seems to be similar to that in DNA solution.     

Thermal stability of soluble mitochondrial H+-ATPase

ATPase melting has been studied by circular dichroism and differential scanning microcalorimetry. Decomposition of the -helix of H⁺-ATPase (in which about 80% of the peptide groups of the enzyme are involved) following thermal treatment is shown to proceed gradually, beginning with room temperature. Effect of nucleotides upon melting is detected in the range of 20–40 C. Above 40 C, the pattern of thermal decomposition of the three-dimensional structure of H⁺-ATPase is independent of the nature of nucleotides present. Highly stable -helical sites have been found in the enzyme molecule. Possible mechanism of formation of such sites is discussed, and the results obtained are compared with data on thermal stability of ATPase from thermophilic bacteria. Structural changes in the molecule following thermal treatment are compared with ATPase activity changes under similar experimental conditions.     

Variations cardio-respiratoires et metaboliques à l'effort sous-maximal (P=40% Pmax) et maximal (Vita Max) en conditions thermiques froide (0‡C) et neutre (20‡C)

Resume Cette étude a pour but d'étudier les variations physiologiques et métaboliques tant au repos, qu'au cours d'efforts physiques de longue durée (2h) à 40% de la puissance maximale et d'efforts exhaustifs dans des conditions d'hypothermie ambiante (0C) en comparaison de valeurs recueillies en conditions thermiques neutres (20C). L'ensemble des épreuves furent exécutées en chambre climatique par: d'une part 11 sujets mâles pour l'effort exhaustif et d'autre part 8 sujets mâles pour l'exercice sous-maximal. Nous avons observé une légère bradycardie à 0C pour nos deux groupes à la fin de l'effort. La VO₂ et la ventilation pulmonaire sont plus élevées à 0C lors de l'effort maximal, nous observons toutefois le phénomène inverse lors de l'exercice sousmaximal. Les concentrations plasmatiques en acide lactique sons moins élevées et les bicarbonates plus élevés en ambiance thermique froide pour les deux conditions expérimentales. Le pH et la température rectale demeurent stables. Le RQ ne présente pas de différences significatives à la fin de chacun des deux efforts à 20C et 0C.

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The purpose of this study is to investigate the responses on cardiorespiratory and metabolic level measured at rest, during a long-term (2 h) submaximal exercise (P=40% Pmax) and during an exhaustive exercise (Vita Max) and recovery time in normal (20C) and in hypothermic (0C) conditions. The ergometric exercises were performed by 11 male subjects (Vita Max) and 8 male subjects (submaximal exercise) in a climatic room. We have observed a slight but mostly not significant (P<.05) bradycardia for both groups at the end of the exercise. VO₂ and pulmonary ventilation are higher at the end of the maximal exercise performed at 0C, this phenomenon is reversed for the submaximal exercise. The lactate concentrations are lower at 0C. For the same temperature the bicarbonates are higher. pH and T rect are not different in both experimental conditions and RQ is not significantly affected by temperature variations.

     

Defective remethylation of homocysteine is related to decreased synthesis of coenzymes B2 in thyroidectomized rats

Summary. We investigated the influence of hypothyroidism on homocysteine metabolism in rats, focusing on a hypothetical deficient synthesis of FAD by riboflavin kinases. Animals were allocated in control group (n=7), thyroidectomized rats (n=6), rats with diet deficient in vitamin B2, B9, B12, choline and methionine (n=7), thyroidectomized rats with deficient diet (n=9). Homocysteine was decreased in operated rats (2.6±1.01 vs. 4.05±1.0mol/L, P=0.02) and increased in deficient diet rats (29.56±4.52 vs. 4.05±1.0mol/L, P=0.001), when compared to control group. Erythrocyte-Glutathione-Reductase-Activation-Coefficient (index of FAD deficiency) was increased in thyroidectomized or deficient diet rats (P=0.004 for both). Methylenetetrahydrofolate-reductase and methionine-synthase activities were decreased in thyroidectomized rats but not in those subjected to deficient diet. Cystathionine--synthase was increased only in operated rats. Taken together, these results showed a defective re-methylation in surgical hypothyroidism, which was due in part to a defective synthesis of vitamin B2 coenzymes. This defective pathway was overcompensated by the increased Cystathionine--synthase activity.     

Novel prebiotic systems: Nucleotide oligomerization in surfactant entrapped water pools

Summary Oligomerization of 5-TMP in water pools entrapped by dodecyl-ammonium chloride surfactant aggregates in benzene: hexane in the presence of dicyanodiimide at temperatures ranging from 21°–72° resulted in the formation of linear and cyclic oligonucleotides containing up to pentamers. Effects of temperature, time and surfactants have been examined. Rate constants for the formation of oligomers have been determined at five different temperatures. These data afforded values of H = 11.8 ± 1.9 Kcal mole^–1, S=–53 6 e.u. and G = 27.4 4.0 Kcal mole^–1. Prebiotic significance of these results are discussed.     

Genetic Linkage Maps of the Guppy (Poecilia reticulata): Assignment of RAPD Markers to Multipoint Linkage Groups

Genetic linkage maps of the guppy (Poecilia reticulata) were constructed from independent crosses between the Tuxedo strain and a feral line (Wildtype). Segregation patterns of random amplified polymorphic DNA (RAPD) markers and phenotypic markers were investigated in F₂ offspring of Tuxedo × Wildtype and Wildtype × Tuxedo crosses. Among the 300 and 276 RAPD markers scored for the respective crosses, linkages were identified for 230 and 212, respectively. The Tuxedo × Wildtype and Wildtype × Tuxedo maps spanned 2100 Kosambi centiMorgans (cM_K) and 1900 cM_K, respectively, in 28 linkage groups. Average marker resolution was 10 cM_K. Genome length was estimated at 4410 cM_K and 4060 cM_K for the respective crosses, with an average physical distance of 166 kbp/cM_K. Several RAPD markers were closely linked to or mapped onto the loci for the sex-determining region (SdR), and the sex-linked black caudal-peduncle (Bcp) and red tail (Rdt) genes. These primary linkage maps are the initial step toward the construction of a composite high-density map to facilitate map-based cloning and marker-assisted selection of quantitative trait loci that are essential for the development of comprehensive breeding programs for the guppy.     

Estimation of sweat rate and thermal tolerance of pure Creole and of Limousin × Creole crossbred growing bulls in Guadeloupe (French West Indies)

Four out of 8 Limousin × Creole (Li × Cr) and 4 out of 8 Creole growing bulls were exposed to full sunshine in Guadeloupe (French West Indies), the others being kept under shade. Sweating rate (measured on the back with capsules filled with a dessicator), rectal temperature and respiratory rhythm were measured between 11 a.m. and 3 p.m., as well as climatic environmental data. Rectal temperature of Creole bulls (overall mean: 38.8C) remains unaffected by black globe temperature (TG), while, for Li × Cr bulls, it is well related to TG, reaching 39.5C when TG=39C; at this moment, Li × Cr respiratory rhythm reaches 100 breaths/min, compared with only 70 for Creole animals. The shade mean levels of these two parameters for each genotype are consistently the same (38.7C and 39 breaths/min). Under full sunshine, mean sweat rate is almost the same for the 2 genotypes; under shelter, the crossbred bulls sweat more than the Creole bulls, both shade means being significantly lower than means in the sun. Probably, when shaded, the Li × Cr animals must sweat more to balance a greater heat production, but their maximum sweat ability is not very different than the one of Creole bulls.     

Expression of a chimeric stilbene synthase gene in transgenic wheat lines

A chimeric stilbene synthase (sts)gene was transferred into wheat. Stilbene synthases play a role in the defence against fungal diseases in some plant species (e.g. groundnut or grapevine) by producing stilbenetype phytoalexins like resveratrol. Resveratrol is also claimed to have positive effects to human health. Embryogenic scutellar calli derived from immature embryos of the two commercial German spring wheat cultivars Combi and Hanno were used as target tissue for cotransformation by microprojectile delivery. The selectable marker/reporter gene constructs contained the bargene either driven by the ubiquitinpromoter from maize (pAHC 25, also containing the uidAgene driven by the ubiquitinpromoter), or by the actinpromoter (pDM 302) from rice. The cotransferred plasmid pStil 2 consisted of a grapevine stscoding region driven by the ubiquitin promoter. Eight transgenic Combi and one Hanno T_Oplant were obtained and, except one Combi T_Oplant, found to be cotransformants due to the integration of both the stsgene and the selectable marker or reporter genes. Expression of the stsgene was proven by RTPCR, and, for the first time, by detection of the stilbene synthase product resveratrol by HPLC and mass spectrometry. The stsgene was expressed in four of the seven transgenic Combi T_{_o}plants. Two of the respective T₁progenies segregated in a Mendelian manner were still expressing the gene. Investigations into methylation of the stsgene showed that in three nonexpressing progenies inactivation was paralleled by methylation.