Isolation and purification ofβ-galactosidase ofLactobacillus murinus CNRZ 313

β-Galactosidase has been isolated inLactobacillus murinus CNRZ 313, and its properties have been studied. The enzyme was purified 292-fold by chromatography on Ultrogel ACA 34, DEAE-Sephadex A-50 columns, and by affinity chromatography in agarose-p-aminophenyl-β-d-thiogalactoside. The purified extract exhibited a single band following polyacrylamide gel electrophoresis (PAGE). Molecular weight was estimated to be 170,000 on an Ultrogel ACA 34 column. Maximum enzymatic activity was observed at 45°C and pH 7 in 50 mM phosphate buffer. The Km for ONPG and ONPG + 20 mM of lactose were 480 μM and 870 μM, respectively. The effect of different salts on the enzymatic activity was studied. An inhibitory effect was observed when using 10 mM CaCl₂, and a stimulating effect when using 10 mM MgCl₂. The latter protected the enzyme from thermal denaturation. Among the protective agents belonging to the sulfhydryl group that was tested, mercaptoethanol and dithiotreitol acted as activators, while glutathione and cysteine acted as inhibitors, of the enzymatic activity.p-β-Galactosidase activity was not observed.     

Tomato juice as a protective agent forLactobacillus acidophilus

The extent of protection afforded by tomato juice, to two strains ofLactobacillus acidophilus during heat shock (80°C) in skimmed milk medium was studied. Presence of tomato juice or its fractions assured a higher survival of both cultures. Strain LB₁H₃ was more resistant than strain LACH to heat shock. Sediments gave the highest protective effect, followed by tomato juice and serum.     

Changes in exercise and post-exercise core temperature under different clothing conditions

 This study evaluates the effect of different levels of insulation on esophageal (T _es) and rectal (T _re) temperature responses during and following moderate exercise. Seven subjects completed three 18-min bouts of treadmill exercise (75% VO_2max, 22°C ambient temperature) followed by 30 min of recovery wearing either: (1) jogging shoes, T-shirt and shorts (athletic clothing); (2) single-knit commercial coveralls worn over the athletic clothing (coveralls); or (3) a Canadian Armed Forces nuclear, bacteriological and chemical warfare protective overgarment with hood, worn over the athletic clothing (NBCW overgarment). T _es was similar at the start of exercise for each condition and baseline T _re was ∼0.4°C higher than T _es. The hourly equivalent rate of increase in T _es during the final 5 min of exercise was 1.8°C, 3.0°C and 4.2°C for athletic clothing, coveralls and NBCW overgarment respectively (P<0.05). End-exercise T _es was significantly different between conditions [37.7°C (SEM 0.1°C), 38.2°C (SEM 0.2°C and 38.5°C (SEM 0.2°C) for athletic clothing, coveralls and NBCW overgarment respectively)] (P<0.05). No comparable difference in the rate of temperature increase for T _re was demonstrated, except that end-exercise T _re for the NBCW overgarment condition was significantly greater (0.5°C) than that for the athletic clothing condition. There was a drop in T _es during the initial minutes of recovery to sustained plateaus which were significantly (P<0.05) elevated above pre-exercise resting values by 0.6°C, 0.8°C and 1.0°C, for athletic clothing, coveralls, and NBCW overgarment, respectively. Post-exercise T _re decreased very gradually from end-exercise values during the 30-min recovery. Only the NBCW overgarment condition T _re was significantly elevated (0.3°C) above the athletic clothing condition (P<0.05). In conclusion, T _es is far more sensitive in reflecting the heat stress of different levels of insulation during exercise and post-exercise than T _re. Physiological mechanisms are discussed as possible explanations for the differences in response. Received: 30 June 1998 / Accepted: 19 February 1999     

Water and temperature relations of softrot bacteria: growth and disease development

The effect of water availability and the temperature of the growth substrate on growth and disease development of softrot bacteria were studied using artificial media and plant material. Water availability was measured as the osmotic potential of a solution (ψ_osm) and was assessed for solutions of PEG4000 and KNO₃ as artificial osmotica and for plant tissue of chicory heads. Growth of softrot bacteria was found at water potentials from ψ= -0.12 MPa to ψ= -8.0 MPa but the lag phase of the growth curve increased with decreasing water potential. The relative growth rates of the three softrot pathogens showed a sigmoidal relationship with water potential, the relative growth rates decreasing rapidly at water potentials lower than ψ= -1.5 MPa. The water potential of harvested chicory heads decreased with storage time of the harvested crop but was still within the growth limits for softrot bacteria. In relation to temperature, the relative growth rate of Erwinia carotovora subsp. carotovora (Ecc) was highest at 10°C, of Erwinia carotovora subsp. atroseptica (Eca) at 15°C and of Pseudomonas marginalis (Pm) at 5°C. Chicory heads of two cultivars, Rumba and Salsa, inoculated with Ecc, had a significantly higher disease severity at 30°C (0.72 for Rumba and 0.47 for Salsa) than at lower or higher temperatures. In conclusion, temperature and water availability during forcing of chicory were not factors limiting populations of softrot bacteria. Possibilities for crop protection thus only avail during chicory root storage. During storage a high death rate combined with a low growth rate of the softrot bacteria may result in a decrease of bacterial populations below the minimum densities needed for infection during the forcing of chicory heads.     

Effects of Menstrual Cycle and Room Temperature on Color Preference

This study investigated the effects of menstrual cycle on color preference in nine normally menstruating female subjects. They were instructed to choose their preferred color out of 45 Munsell hues every 5 min at ambient temperatures (T _a) of 28°C (630-800 h), from 28°C to 23°C (800-900 h) and at 23°C (900-930 h). Warmer color hues were preferred during the luteal phase than the follicular phase at 28°C, while there did not exist any significant differences at other T _as. The findings that a preference for warmer colors occurred in the luteal phase at 28°C is discussed in terms of the load error between actual core temperature and its setpoint.     

The Characteristics of Arylamine N-Acetyltransferase in Pseudomonas aeruginosa

N-Acetyltransferase (NAT), responsible for bioactivation and detoxification of arylamines, has been demonstrated to be widely distributed in many organisms ranging from humans to microorganisms. Using high performance liquid chromatography (HPLC) to analyze NAT activity in bacteria, the authors found that Pseudomonas aeruginosa exhibited high NAT activity with 2-aminofluorene (2-AF) as substrate. Characteristics of this bacterial NAT were further investigated. The N-acetylation catalyzed by this enzyme is an acetyl coenzyme A (AcCoA)-dependent reaction. As the concentration of AcCoA in the reaction mixture was increased, the apparent K _m and V _max for 2-AF increased. The K _m and V _max were 0.504 ± 0.056 mM and 31.92 ± 3.23 nmol/min/mg protein, respectively, for the acetylation of 2-AF with 0.5 mM AcCoA. The optimum pH for the enzyme activity was estimated to be around 8.5. It was active at a temperature range from 5°C to 55°C, with maximum activity at 37°C. The enzyme activity was inhibited by divalent metal ions including Cu⁺⁺, Fe⁺⁺, Zn⁺⁺, Ca⁺⁺, Co⁺⁺, Mn⁺⁺, and Mg⁺⁺, suggesting that a sulfhydryl group is involved in the N-acetylation activity. The three chemical modification agents, iodoacetamide, phenylglyoxal, and diethylpyrocarbonate, all exhibited a dose-, time-, and temperature-dependent inhibition effect. Preincubation of the NAT with AcCoA provided significant protection against the inhibition of iodoacetamide and diethylpyrocarbonate, but only partial protection against the inhibition of phenylglyoxal. These results indicate that cysteine, histidine, and arginine residues are essential for this bacterial enzyme activity, and the first two are likely to reside on the AcCoA binding site, but arginine residue may be located only near the AcCoA binding site. Our data demonstrate that P. aeruginosa possesses highly active N-acetyltransferase which shares a similar catalytic mechanism as that of higher organisms. These findings are very helpful for further investigating the role of arylamine NAT in this bacterial species. Received: 29 August 1997 / Accepted: 23 December 1997     

meso-α,ɛ-Diaminopimelate d-Dehydrogenase: Sulfhydryl Group Modification

meso-α,?-Diaminopimelate D-dehydrogenase was inhibited by sulfhydryl reagents such as p-chloromercuribenzoate and HgCl₂. Two sulfhydryl groups were titrated per molecule in the presence and absence of 6 M guanidine hydrochloride: the enzyme contained one sulfhydryl group per subunit. Modification of the sulfhydryl groups with p-chloromercuribenzoate, 5,5'-dithiobis(2-nitrobenzoic acid), 4,4'-dithiopyridine, N-ethylmaleimide, and iodoacetic acid was accompanied by a loss of enzyme activity. However, modification of sulfhydryl groups of the enzyme with cyanide did not affect the activity. Thus, the introduction of bulky or charged substituents to sulfhydryl groups decreased the catalytic activity of the enzyme, but modification of the groups with the small and uncharged group, a cyano group, did not. The sulfhydryl groups did not play an essential role in catalysis.     

Enhancing viability of two biocontrol yeasts in liquid formulation by applying sugar protectant combined with antioxidant

Survivals of Cryptococcus laurentii and Pichia membranaefaciens in liquid formulations with sugar protectants (trehalose and galactose) and L-ascorbic acid (Vc) were investigated during storage at 4°C and 25°C. When galactose or trehalose was used alone as protectant, C. laurentii maintained relatively high viability in potassium phosphate buffer. Addition of Vc to trehalose improved its protective effect. P. membranaefaciens maintained viability >60% after 90 days at 4°C when 5% galactose served as a protectant, and its combination with Vc was the most effective at maintaining viability. Moreover, liquid formulation kept higher viability of the two yeasts at 4°C than at 25°C. Biocontrol efficiency of the two yeasts was maintained after formulation and storage. The results indicate that trehalose is considered as a suitable protectant for liquid formulation of C. laurentii, while galactose is better for P. membranaefaciens. Combining Vc with the sugars improves the protective efficiency.     

Plant responses to H2S and SO2 fumigation. I. Effects on growth, transpiration and sulfur content of spinach

Exposure of spinach (Spinacia oleracea L. cv. Monosa) to 0.25 μl l_?1 H₂S reduced the relative growth rate by 26, 47 and 60% at 15, 18 and 25°C, respectively. Shoot to root ratio decreased in plants fumigated at 18 and 25°C. Growth of spinach was not affected by a 2-week exposure to 0.10 or 0.25 μl l_?1 SO₂. Both H₂S and SO₂ fumigation increased the content of sulfhydryl compounds and sulfate. A 2-week exposure to 0.25 μl l_?1 H₂S resulted in an increase in sulfhydryl and sulfate content of 250 to 450% and 63 to 248% in the shoots, respectively, depending on growth temperature. Exposure to 0.15 and 0.30 μl l_?1 H₂S at 20°C for 2 weeks resulted in a 46% increase in sulfate content of the shoots at 0.30 μl l_?1 and no detectable increase at 0.15 μl l_?1 H₂S; the sulfate content of the roots increased by 195 and 145% at 0.15 and 0.30 μl l_?1 H₂S, respectively. Fumigation with 0.25 μl l_?1 SO₂ at 20°C for 2 weeks resulted in an increase in sulfhydryl content and sulfate content in the shoots of 285% and 300 to 1100%. H₂S fumigation during the 12 h light period or only during the dark period resulted in identical growth reduction and accumulation of sulfhydryl compounds; they were about 50 and 67% of those observed in continuously exposed plants. H₂S- and SO₂-exposed plants showed an increased transpiration rate, which was mainly caused by an increased dark-period transpiration. No effect of H₂S and SO₂ on the water uptake of the plants and the osmotic potential of the leaves was detected. Plants fumigated with 0.25 μl l_?1 H₂S for 2 weeks were smaller and differed morphologically from the control plants by slightly more abaxially curved leaf margins. Cross sections of the leaves showed smaller cells at the margins and smaller and fewer air spaces. The increased transpiration in the H₂S-exposed plants is discussed in relation to the observed morphological changes.     

Trehalose protects wine yeast against oxidation under thermal stress

Accumulation of trehalose in yeasts has been suggested to be an important mechanism of tolerance against adverse stress conditions, particularly in thermal stress. However, under thermal stress, it is not clear if the mechanism of protection is related to its antioxidant role. In this study, a newly isolated wine yeast Saccharomyces cerevisia was used to examine the protective effect of trehalose against oxidation during thermal stress treatment. Cells were treated either with a mild heat treatment at 37°C (which leads to trehalose accumulation) or with a 50 mM trehalose solution and then exposed to a high temperature of 53°C. According to our results, mild heat treatment at 37°C and trehalose addition which promote accumulation of trehalose significantly increased cell survival upon exposure to thermal stress at 53°C which seems to be correlated with decrease in reactive oxygen species levels and lipid peroxidation. Trehalose could protect yeast from oxidative injuries under thermal stress.     

Effects of holding temperature and handling stress on the upper thermal tolerance of threadfin shad Dorosoma petenense

The critical thermal maxima (T_MAX) of threadfin shad Dorosoma petenense exposed to standardized stress (30 s handling in a dip‐net), simulating stressors endured during fish loading before transport, were measured over a range of holding temperatures (15, 20 and 25° C). Dorosoma petenense T_MAX showed a significant thermal effect, displaying mean ±s.d . critical thermal maxima of 26·5 ± 1·6, 30·9 ± 1·2 and 33·3 ± 1·4° C, when tested at temperatures of 15, 20 and 25° C, respectively. Dorosoma petenense T_MAX levels were also affected by stress, with handled fish showing significantly lower values than control fish exposed to 15 (mean ±s.d . T_MAX = 25·6 ± 2·0° C), 20 (27·6 ± 2·8° C) and 25° C (32·0 ± 2·6° C). In addition to providing basic information on D. petenense thermal tolerance, experimental results suggest that fishery managers should consider the whole suite of potential stressors, such as air exposure during handling and fish loading, when developing management criteria.     

Comparative NMR studies of diffusional water permeability of red blood cells from different species: XVI Dingo (Canis familiaris dingo) and dog (Canis familiaris)

As part of a programme of comparative measurements of P _d (diffusional water permeability) the RBCs (red blood cells) from dingo (Canis familiaris dingo) and greyhound dog (Canis familiaris) were studied. The morphologies of the dingo and greyhound RBCs [examined by light and SEM (scanning electron microscopy)] were found to be very similar, with regard to aspect ratio and size; the mean diameters were estimated to be the same (～7.2 μm) for both dingo and greyhound RBCs. The water diffusional permeability was monitored by using an Mn²⁺‐doping ¹H NMR technique at 400 MHz. The P _d (cm/s) values of dingo and greyhound RBCs were similar: 6.5×10^?3 at 25°C, 7.5×10^?3 at 30°C, 10×10^?3 at 37°C and 11.5×10^?3 at 42°C. The inhibitory effect of a mercury‐containing SH (sulfhydryl)‐modifying reagent PCMBS (p‐chloromercuribenzene sulfonate) was investigated. The maximal inhibition of dingo and greyhound RBCs was reached in 15–30 min at 37°C with 2 mmol/l PCMBS. The values of maximal inhibition were in the range 72–74% when measured at 25°C and 30°C, and ～66% at 37°C. The lowest value of P _d (corresponding to the basal permeability to water) was ～2–3×10^?3 cm/s in the temperature range 25–37°C. The E _a,d (activation energy of water diffusion) was 25 kJ/mol for dingo RBC and 23 kJ/mol for greyhound RBCs. After incubation with PCMBS, the values of E _a,d increased, reaching 46–48 kJ/mol in the condition of maximal inhibition of water exchange. The electrophoretograms of membrane polypeptides of the dingo and greyhound RBCs were compared and seen to be very similar. We postulate that the RBC parameters reported in the present study are characteristic of all canine species and, in particular in the two cases presented here, these parameters have not been changed by the peculiar Australian habitat over the millennia (as in the case of the dingo) or over shorter time periods, decades or centuries (as in the case of the domestic greyhound).     

Energetics of arousal episodes in hibernating arctic ground squirrels

Arctic ground squirrels overwintering in northern Alaska experience average soil temperature of −10°C. To examine energetic costs of arousing from hibernation under arctic compared to temperate conditions, captive ground squirrels were maintained in ambient temperatures (T _a) of 2, −5 and −12°C. Rates of oxygen consumption and carbon dioxide production were used to estimate metabolic rate and fuel use during the three phases of arousal episodes: rewarming, euthermia, and recooling. Respiratory quotient comparisons suggest exclusive use of lipid during rewarming and mixed fuel use during euthermia. Animals rewarming from torpor at T _a −12°C took longer, consumed more oxygen, and attained higher peak rates of oxygen consumption when compared to 2°C. T _a had no significant effect on cost or duration of the euthermic phase. Animals recooled faster at −12°C than at 2°C, but total oxygen consumption was not different. T _a had no significant effect on the total cost of arousal episodes when all three phases are included. Arousal episodes account for 86% of estimated costs of a complete hibernation cycle including torpor when at 2°C and only 23% at −12°C. Thus, due to the higher costs of steady-state metabolism during torpor, proportional metabolic costs of arousal episodes at T _a characteristic of the Arctic are diminished compared to relative costs of arousals in more temperate conditions.     

Bcl‐xL overexpression does not enhance specific erythropoietin productivity of recombinant CHO cells grown at 33°C and 37°C

Overexpression of bcl‐xL in recombinant Chinese hamster ovary (rCHO) cells has been known to suppress apoptotic cell death and thereby extend culture longevity during batch culture. However, its effect on specific productivity (q) of rCHO cells is controversial. This study attempts to investigate the effect of bcl‐xL overexpression on q of rCHO cells producing erythropoietin (EPO). To regulate the bcl‐xL expression level, the Tet‐off system was introduced in rCHO cells producing EPO (EPO‐off‐bcl‐xL). The bcl‐xL expression level was tightly controlled by doxycycline concentration. To evaluate the effect of bcl‐xL overexpression on specific EPO productivity (q_EPO) at different levels, EPO‐off‐bcl‐xL cells were cultivated at the two different culture temperatures, 33°C and 37°C. The q_EPO at 33°C and 37°C in the presence of 100 ng/mL doxycycline (without bcl‐xL overexpression) were 4.89 ± 0.21 and 3.18 ± 0.06 μg/10⁶cells/day, respectively. In the absence of doxycycline, bcl‐xL overexpression did not affect q_EPO significantly, regardless of the culture temperature, though it extended the culture longevity. Taken together, bcl‐xL overexpression showed no significant effect on the q_EPO of rCHO cells grown at 33°C and 37°C. © 2009 American Institute of Chemical Engineers Biotechnol. Prog., 2009     

Experimental evidence for the attenuating effect of SOM protection on temperature sensitivity of SOM decomposition

The ability to predict C cycle responses to temperature changes depends on the accurate representation of temperature sensitivity (Q₁₀) of soil organic matter (SOM) decomposition in C models for different C pools and soil depths. Theoretically, Q₁₀ of SOM decomposition is determined by SOM quality and availability (referred to here as SOM protection). Here, we focus on the role of SOM protection in attenuating the intrinsic, SOM quality dependent Q₁₀. To assess the separate effects of SOM quality and protection, we incubated topsoil and subsoil samples characterized by differences in SOM protection under optimum moisture conditions at 25 °C and 35 °C. Although lower SOM quality in the subsoil should lead to a higher Q₁₀ according to kinetic theory, we observed a much lower overall temperature response in subsoil compared with the topsoil. Q₁₀ values determined for respired SOM fractions of decreasing lability within the topsoil increased from 1.9 for the most labile to 3.8 for the least labile respired SOM, whereas corresponding Q₁₀ values for the subsoil did not show this trend (Q₁₀ between 1.4 and 0.9). These results indicate the existence of a limiting factor that attenuates the intrinsic effect of SOM quality on Q₁₀ in the subsoil. A parallel incubation experiment of ¹³C‐labeled plant material added to top‐ and subsoil showed that decomposition of an unprotected C substrate of equal quality responds similarly to temperature changes in top‐ and subsoil. This further confirms that the attenuating effect on Q₁₀ in the subsoil originates from SOM protection rather than from microbial properties or other nutrient limitations. In conclusion, we found experimental evidence that SOM protection can attenuate the intrinsic Q₁₀ of SOM decomposition.     

Expression of heat shock proteins in response to high and low temperature extremes in diapausing pharate larvae of the gypsy moth,Lymantria dispar

Diapausing pharate first instars of the gypsy moth, Lymantria dispar, respond to high temperature (37–41°C) by suppressing normal protein synthesis and synthesizing a set of seven heat shock proteins with M_rs of 90,000, 75,000, 73,000, 60,000, 42,000, 29,000, and 22,000 as determined by SDS-PAGE. During recovery at 25°C from heat shock, synthesis of the heat shock proteins gradually decreases over a period of 6 h, while normal protein synthesis is restored. A subset of these same heat shock proteins is also expressed during recovery at 4°C or 25°C from brief exposures to low temperature (-10 to 20°C), and its expression is more intense with increased severity of cold exposure. During recovery at 4°C after 24 h at ?20°C, both 90,000 and 75,000 M_r heat shock proteins are expressed for more than 96 h. While normal protein synthesis is suppressed during heat shock and recovery from heat shock, normal protein synthesis coincides with synthesis of the heat shock proteins during recovery from low temperatures, thus implying that expression of the heat shock proteins is not invariably linked to suppression of normal protein synthesis. Western transfer, using a monoclonal antibody that recognizes the inducible form of the human 70,000 M_r heat shock protein, demonstrates that immunologically related proteins in the gypsy moth are expressed at 4°C and during recovery from cold and heat shock.     

Optimisation of the spray drying process of formulating the post-harvest biocontrol agent Meyerozyma caribbica

In this study, conditions of the spray-drying process of Meyerozyma caribbica were optimised using response surface methodology (RSM). The effect of three parameters (protective agent, inlet air temperature and protective agent concentration) on the cell viability of Meyerozyma caribbica was evaluated. Each parameter was evaluated at three levels. All of the evaluated factors presented an effect on the viability of the control agent. According to RSM, optimal conditions include the use of trehalose with a concentration of 7.75% (w/v) and inlet air temperature of 112.5°C. The validation of the optimal spray-drying conditions allows obtaining a formulation of M. caribbica with 95.41?±?0.93% viability, 5?±?0.37% humidity and a_w of 0.33?±?0.11. Storage for six months at 4°C presented a 5% loss in cell viability.     

Characterizing the ability of an ice recrystallization inhibitor to improve platelet cryopreservation

Improving aspects of platelet cryopreservation would help ease logistical challenges and potentially expand the utility of frozen platelets. Current cryopreservation procedures damage platelets, which may be caused by ice recrystallization. We hypothesized that the addition of a small molecule ice recrystallization inhibitor (IRI) to platelets prior to freezing may reduce cryopreservation-induced damage and/or improve the logistics of freezing and storage. Platelets were frozen using standard conditions of 5–6% dimethyl sulfoxide (Me₂SO) or with supplementation of an IRI, N-(2-fluorophenyl)-d-gluconamide (2FA), prior to storage at −80 °C. Alternatively, platelets were frozen with 5–6% Me₂SO at −30 °C or with 3% Me₂SO at −80 °C with or without 2FA supplementation. Supplementation of platelets with 2FA improved platelet recovery following storage under standard conditions (p = 0.0017) and with 3% Me₂SO (p = 0.0461) but not at −30 °C (p = 0.0835). 2FA supplementation was protective for GPVI expression under standard conditions (p = 0.0011) and with 3% Me₂SO (p = 0.0042). Markers of platelet activation, such as phosphatidylserine externalization and microparticle release, were increased following storage at −30 °C or with 3% Me₂SO, and 2FA showed no protective effect. Platelet function remained similar regardless of 2FA, although functionality was reduced following storage at −30 °C or with 3% Me₂SO compared to standard cryopreserved platelets. While the addition of 2FA to platelets provided a small level of protection for some quality parameters, it was unable to prevent alterations to the majority of in vitro parameters. Therefore, it is unlikely that ice recrystallization is the major cause of cryopreservation-induced damage.     

Maximum rates of sustained metabolic rate in cold-exposed Djungarian hamsters (Phodopus sungorus): the second wind

Djungarian hamsters (Phodopus sungorus) tolerate short-term exposure to ambient temperatures (T _as) down to −70°C, but surprisingly, previously appeared to reach maximum sustainable metabolic rate (SusMR) when kept at T _as as high as ≥−2°C. We hypothesized that SusMR in Djungarian hamsters may be affected by the degree of prior cold acclimation and temporal patterns of T _a changes experienced by the animals, as average T _a declines. After cold-acclimation at +5°C for 6 weeks, hamsters reached rates of SusMR that were 35% higher than previously determined and were able to maintain positive energy balances down to T _a −9°C. SusMR was unaffected, however, by whether mean cold load was constant or caused by T _as cycling between +3°C and as low as −25°C, at hourly intervals. At mean T _as between +3 and −3°C hamsters significantly reduced body mass and energy expenditure, but were able to maintain stable body mass at lower T _as (−5 to −9°C). These results indicate that prior cold-acclimation profoundly affects SusMR in hamsters and that body mass regulation may play an integral part in maintaining positive energy balance during cold exposure. Because the degree of instantaneous cold load had no effect on SusMR, we hypothesize that limits to energy turnover in Djungarian hamsters are not determined by the capacity to withstand extreme temperatures (i.e., peripheral limits) but are due to central limitation of energy intake.     

Purification and properties of the endo-1,4-β-glucanase III from Ruminococcus albus

Endoglucanase III (EGIII) was purified from Ruminococcus albus culture supernatant. An enzyme having a molecular weight of 53,000 was stabilized by mercaptoethanol and inhibited by sulfhydryl group-blocking reagents, and exhibited its highest CMC-degrading activity of pH 5.7 and 55°C. The enzyme hydrolyzed cellobiose (G2) and cellotriose (G3) only negligibly, but significantly hydrolyzed cellotetraose (G4), cellopentaose (G5) and cellohexaose (G6). The major hydrolysis reactions conducted by the enzyme were G4→2G2, G5→G2+G3, G6→G2+G4 and G6→2G3. The V_max values of these reactions increased remarkably while the K_m values decreased significantly with an increase in degree of polymerization of the substrate.