Maximum voluntary temperature of insect larvae reveals differences in their thermal biology

We investigate the thermoregulatory behaviors of larvae of four species of Drosophila (D. melanogaster, D. subobscura, D. pseudoobscura, and D. mojavensis), a thermotolerant strain of Drosophila melanogaster (T strain) known to differ in thermal biology, and two mutant stocks of D. melanogaster that have (as adults) defective thermoregulatory behavior. We describe and evaluate new techniques to measure two indices of maximum voluntary temperature of insect larvae. Both measures were highly repeatable within lines (species, strains, or mutants). One measure (temperature at which larvae stood upright) differed among lines consistent with expectations based on adult thermal ecology, suggesting that this measure will be useful measures of thermoregulatory set-points of larvae. The second measure (temperature of emergence from media) is less discriminatory.     

Crosslinked enzyme crystals of glucoamylase as a potent catalyst for biotransformations

Glucoamylase (E.C: 3.2.1.3, alpha-(1-->4)-glucan glucohydrolase) mainly hydrolyzes starch and has been extensively used in the starch, glucose (dextrose), and fermentation industries. Immobilized glucoamylase has an inherent disadvantage of lower conversion rates and low thermostability of less than 55 degrees C when used in continuous operations. We have developed crosslinked enzyme crystals (CLEC) of glucoamylase that overcome the above disadvantages, possess good thermal stability and retain 98.6% of their original activity at 70 degrees C for 1h, 77% activity at 80 degrees C for 1h, and 51.4% activity at 90 degrees C for 0.5h. CLEC glucoamylase has a specific activity of 0.0687 IU/mg and a yield of 50.7% of the original activity of the enzyme under optimum conditions with starch as the substrate. The crystals obtained are rhombohedral in shape having a size approximately 10-100 microm, a density of 1.8926 g/cm(3) and a surface area of 0.7867 m(2)/g. The pH optimum of the glucoamylase crystals was sharp at pH 4.5, unlike the soluble enzyme. The kinetic constants V(max) and K(m) exhibited a 10-fold increase as a consequence of crystallization and crosslinking. The continuous production of glucose from 10% soluble starch and 10% maltodextrin (12.5 DE) by a packed-bed reactor at 60 degrees C had a productivity of 110.58 g/L/h at a residence time of 7.6 min and 714.1g/L/h at a residence time of 3.4 min, respectively. The CLEC glucoamylase had a half-life of 10h with 4% starch substrate at 60 degrees C.     

Thermal ecophysiology of a native and an invasive gecko species in a tropical dry forest of Mexico

For ectotherms, thermal physiology plays a fundamental role in the establishment and success of invasive species in novel areas and, ultimately, in their ecological interactions with native species. Invasive species are assumed to have a greater ability to exploit the thermal environment, higher acclimation capacities, a wider thermal tolerance range, and better relative performance under a range of thermal conditions. Here we compare the thermal ecophysiology of two species that occur in sympatry in a tropical dry forest of the Pacific coast of Mexico, the microendemic species Benedetti's Leaf-toed Gecko (Phyllodactylus benedettii) and the invasive Common House Gecko (Hemidactylus frenatus). We characterized their patterns of thermoregulation, thermoregulatory efficiency, thermal tolerances, and thermal sensitivity of locomotor performance. In addition, we included morphological variables and an index of body condition to evaluate their effects on the thermal sensitivity of locomotor performance in these species. Although the two species had similar selected temperatures and thermal tolerances, they contrasted in their thermoregulatory strategies and thermal sensitivity of locomotor performance. Hemidactylus frenatus had a higher performance than the native species, P. benedettii, which would represent an ecological advantage for the former species. Nevertheless, we suggest that given the spatial and temporal limitations in habitat use of the two species, the probability of agonistic interactions between them is reduced. We recommend exploring additional biotic attributes, such as competition, behavior and niche overlap in order assess the role of alternative factors favoring the success of invasive species.     

The role of temperature in determining distributions and coexistence of three species of Plestiodon

The Common Five-Lined Skink (Plestiodon fasciatus), Southeastern Five-Lined Skink (P. inexpectatus), and Broadheaded Skink (P. laticeps) are all found in forested ecosystems of the southeastern United States and occur in apparent sympatry. No aspect of their ecology has been quantitatively shown to be different. Differences in their natural history, such as northern extent of geographic range, observations of differences in habitat preference, and differing physiological responses to temperature, implicate the thermal environment as a potential niche dimension by which these organisms partition resources. Here we show that the northernmost extent of each species' range can be adequately discriminated by temperature extremes and that their preferred habitats differ in canopy cover, mean temperature, and daily maximum temperatures. These differences are supported by observations in the field. Based upon these findings and observations, we conclude that these species may be able to exist in such close proximity due to differences in their thermal niche, particularly the two like-sized species, P. fasciatus and P. inexpectatus. P. laticeps is a much larger lizard as an adult and functionally coexists with its congeners, potentially by partitioning resources along a different axis.     

Keeping warm in the cold: On the thermal benefits of aggregation behaviour in an intertidal ectotherm

Temperature is a primary determinant for species geographic ranges. In the context of global warming, most attention focuses upon the potential effects of heat stress on the future distribution of ectothermic species. Much less attention has, however, been given to cold thermal stress although it also sets species thermal window limits, hence distribution ranges. This study was conducted in winter on a South-Australian rocky shore in order to investigate the potential thermal benefits of the aggregation behavior observed in the dominant gastropod Nerita atramentosa. Thermal imaging was used to measure the body temperatures of 3681 aggregated individuals and 226 solitary individuals, and surrounding substratum temperature. N. atramentosa aggregates and solitary individuals were significantly warmer than their surrounding substratum. The temperature deviation between aggregates and substratum was, however, ca. 2 °C warmer than the one observed between solitary individuals and substratum. This result is critical since a body temperature increase of only a few degrees might enhance individual performance, hence organismal fitness, and could potentially drive changes in interspecific relationships. Besides, the potential higher thermal inertia of aggregates might increase the snail adaptive ability to abrupt environmental changes. We further investigate the potential thermal heterogeneity within an aggregate in order to identify any thermally advantageous position. Patch centers are significantly warmer than their edges, hence snails experience greater thermal advantages in the aggregate center. Finally, we examined the potential effect of aggregate size on snail temperature and thermal spatial heterogeneity. We identified an aggregate size threshold (216 individuals) beyond which all snails had equal thermal benefits, regardless of their spatial positions within an aggregate. While the determinism of this aggregate size threshold requires further investigations, the present work uniquely identified the thermal benefits of aggregation behavior for intertidal ectotherms under cold weather conditions. The implications of the present finding are discussed in the general framework of the ability of ectothermic populations to face environmental changes.     

Thermal and guanidine hydrochloride-induced denaturation of human cystatin C

Wild-type human cystatin C is directly involved in pathological fibrils formation, leading to hemorrhage, dementia and eventually death of people suffering from cerebral amyloid angiopathy. Some studies on cystatin C oligomerization have been already done but some points are still unclear. In order to learn more about this important process, we have investigated thermal and chemical (guanidine hydrochloride-induced) denaturation of human cystatin C. Studies performed using tryptophan fluorescence, calorimetry, circular dichroism and Fourier transform infrared spectroscopy demonstrate that neither chemical nor thermal denaturation of hCC are simple two-state events. One recognized intermediate form was dimeric cystatin C, whose appearance was preceded mainly by changes in the L2 binding loop. The other form occurred only in the chemical denaturation process and was characterized by partially recovered interactions maintaining the protein tertiary structure. Our studies also strongly indicate that the -structural motif of cystatin C is directly implicated in formation of temperature-induced aggregates.Abbreviations Gdn.HCl guanidine hydrochloride - hCC human cystatin C     

Preparation, spectral characterization, cytotoxic and thermal studies of platinum (IV) thiohydrazone complexes

Platinum (IV) complexes of type [Pt(L)₂Cl₂] [where, L=benzaldehyde-N,N-diphenyl-N-thiohydrazone (L¹), salicylaldehyde-N,N-diphenyl-N-thiohydrazone (L²), acetaphenone-N,N-diphenyl-N-thiohydrazone (L³) and cyclohexanone-N,N-diphenyl-N-thiohydrazone (L⁴)] have been synthesized. The thiohydrazones can exist as thione–thiol tautomer and coordinates as a bidentate N–S ligand. The ligands found to act in monobasic bidentate fashion. Analytical data reveals that metal to ligand stoichiometry is 1:2. The complexes have been characterized by elemental analysis, IR, mass, electronic, ¹H NMR spectroscopic studies, and TG/DTA study. Various kinetic and thermodynamic parameters like order of reaction (n), activation energy (E_a), apparent activation entropy (S^#) and heat of reaction (ΔH) have also been carried out for one complex. Cytotoxic study has also been carried out for one complex.     

Thermally stable and hydrogen peroxide tolerant manganese peroxidase (MnP) from Lenzites betulinus

A thermally stable and hydrogen peroxide tolerant manganese peroxidase (MnP) was purified from the culture medium of Lenzites betulinus by ion exchange chromatography, gel filtration and isoelectric focusing chromatography. The MnP purified from L. betulinus (L-MnP) has a molecular mass of 40 kDa and its isoelectric point was determined to be 6.2. The first 19 amino acids at the N-terminal end of the L-MnP sequence were found to exhibit 74% identity with those of a Phlebia radiata MnP. L-MnP was proved to have the highest hydrogen peroxide tolerance among MnPs reported so far. It retained more than 60% of the initial activity after thermal treatment at 60°C for 60 min, and also retained more than 60% of the initial activity after exposure to 10 mM hydrogen peroxide for 5 min at 37°C.     

Stabilization of G-quadruplex structure on vascular endothelial growth factor gene promoter depends on CpG methylation site and cation type

Background

DNA methylation at the 5-position of cytosine is an epigenetic modification of CpG dinucleotides. In addition to CpG methylation, the G-quadruplex (G4) structure has been reported as a regulator of gene expression. The identification of G4 forming sequences in CpG islands suggests an involvement of CpG-methylated G4 structures in biological processes; however, few reports have addressed the effects of CpG methylation on G4 structure.

If space is provided,bulky modification on the rim of azurin's beta-barrel results in folded protein

Pseudomonas aeruginosa azurin is a blue-copper protein with a beta-barrel fold. Here we report that, at conditions where thermal unfolding of apo-azurin is reversible, the reaction occurs in a single step with a transition midpoint (T(m)) of 69 degrees C (pH 7). The active-site mutation His117Gly creates a cavity in the beta-barrel near the surface but does not perturb the overall fold (T(m) of 64 degrees C, pH 7). Oxidation of the active-site cysteine (Cysteine-112) in wild-type azurin, which occurs readily at higher temperatures, results in a modified protein that cannot adopt a native-like structure. In sharp contrast, Cysteine-112 oxidation in His117Gly azurin yields a modified apo-azurin that appears folded and displays cooperative, reversible unfolding (T(m) approximately 55 degrees C, pH 7). We conclude that azurin's beta-barrel is a rigid structural element that constrains the structure of its surface; a bulky modification can only be accommodated if complementary space is provided.