Deciphering the structural role of histidine 83 for heme binding in hemophore HasA

Heme carrier HasA has a unique type of histidine/tyrosine heme iron ligation in which the iron ion is in a thermally driven two spin states equilibrium. We recently suggested that the H-bonding between Tyr75 and the invariantly conserved residue His83 modulates the strength of the iron-Tyr75 bond. To unravel the role of His83, we characterize the iron ligation and the electronic properties of both wild type and H83A mutant by a variety of spectroscopic techniques. Although His83 in wild type modulates the strength of the Tyr-iron bond, its removal causes detachment of the tyrosine ligand, thus giving rise to a series of pH-dependent equilibria among species with different axial ligation. The five coordinated species detected at physiological pH may represent a possible intermediate of the heme transfer mechanism to the receptor.     

Metabolic Effects of Mental Stress during Over- and Underfeeding in Healthy Women

Objective: To assess the short-term consequences of carbohydrate or fat overfeeding or of food restriction on the metabolic effects of mental stress in healthy lean women. Research Methods and Procedures: The effects of a sympathetic activation elicited by mental stress were evaluated in a group of healthy women after standardized isocaloric feeding (ISO) or after a 3-day overfeeding with 40% excess calories as either carbohydrate overfeeding (CHO OF) or fat overfeeding (FAT OF). Oxygen consumption rate (VO ₂) was measured as an index of energy expenditure, and subcutaneous glycerol concentrations were monitored with microdialysis. The same measurements were performed in another group of healthy women after ISO and after a 3-day period of underfeeding with a protein sparing modified fast (UF). Results: In all conditions, mental stress significantly increased heart rate, blood pressure, plasma norepinephrine and epinephrine concentrations, and VO ₂, and produced a nonsignificant increase in subcutaneous glycerol concentrations. CHO OF and FAT OF did not alter the effects of mental stress on VO ₂ and subcutaneous glycerol concentrations. In contrast, UF increased basal VO ₂ but significantly reduced its stimulation by mental stress. UF also enhanced the increase in subcutaneous glycerol concentrations during mental stress. Discussion: UF reduces the stimulation of energy expenditure and enhances lipolysis during sympathetic activation. These adaptations may be involved in mobilization of endogenous fat while limiting weight loss. In contrast, short-term overfeeding fails to alter the sympathetic control of energy expenditure and lipolysis.     

Energy Metabolism in Rhizomes of Acorus calamus (L.) and in Tubers of Solanum tuberosum (L.) With Regard to their Anoxia Tolerance

Rhizomes of the marsh plant Acorus calamus (L.) and tubers of the flooding-intolerant Solanum tuberosum (L.) var. Bintje, both kept under strict anoxia, differ markedly in their fermentation properties. The fermentation capacities as measured by ADH and LDH activities and their respective product concentrations were estimated. While rhizomes of Acorus calamus, having high ADH and low LDH activities, accumulate mainly ethanol, tubers of Solanum tuberosum tend towards lactic acid fermentation. The total amount of adenine nucleotides is quite stable in Acorus calamus, whereas they show a sharp decline in S. tuberosum during the first 6h of anoxia. The adenylate energy charge of A. calamus recovers after a short initial drop (AEC > 0.8). AEC values of S. tuberosum decrease rapidly and remain at very low values (AEC ～ 0.3). Tuber tissues became soft and lost viability after about 48–72 h of anoxia at 25 °C. This might be due to tissue acidification and impaired energy metabolism, but not to the lack of energy reserves. Energy metabolism of A. calamus is well adapted to anoxia.     

Seasonal variation and an “outbreak” of frog predation by tamarins

We report temporal variation and an “outbreak” of frog predation by moustached tamarins, Saguinus mystax, in north-eastern Peruvian Amazonia. Frog predation rates were generally very low, but strongly increased in October 2015. Other high rates, identified by outlier analyses, were also observed in September–November of other years. Over all study years, predation rates in this 3-month period were significantly higher than those in the remainder of the year, suggesting a seasonal pattern of frog predation by tamarins. Reduced fruit availability or increased frog abundance or a combination of both may be responsible for both the seasonal pattern and the specific “outbreak” of frog predation.     

Effect of plant growth regulators on indirect shoot organogenesis of Ficus religiosa through seedling derived petiole segments

Ficus religiosa is known as a long-lived multipurpose forest tree. The tree plays an important role for religious, medicinal, and ornamental purposes. However, the propagation rate of Ficus religiosa is low in natural habitat so the plant tissue culture techniques are an applicable method for multiplication of this valuable medicinal plants. Thus, the aim of this study is to understand the effect of different auxin/cytokinin ratios on indirect shoot organogenesis of this plant. According to our results, the maximum callus induction frequency (100%) was obtained on Murashige and Skoog (MS) medium supplemented with 0.5?mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) plus 0.05?mg/l 6-benzylaminopurine (BAP) from petiole segments. For shoot induction purpose, the yellow-brownish, friable, organogenic calli were inoculated on shoot induction medium. On MS medium supplemented with 1.5?mg/l BAP and 0.15?mg/l Indole-3-butyric acid (IBA), 96.66% of the petiole-derived calli responded with an average number of 3.56 shoots per culture. The highest root formation frequency (96.66%), root number (5.5), and root length (4.83?cm) were achieved on MS medium containing 2.0?mg/l IBA plus 0.1?mg/l Naphthaleneacetic acid (NAA). The rooted shoots were successfully transferred to field condition and the substrate with the mixture of cocopeat and perlite (1:1) had the highest survival rate (96.66%). This is the first report of an effective in vitro organogenesis protocol for F. religiosa by indirect shoot organogenesis through axenic seedling derived petiole explants, which can be efficiently employed for conservation of this important medicinal plant species as well as the utilization of active biomolecules.     

An investigation into the role of different constituents in damage accumulation in arterial tissue and constitutive model development

Carotid atherosclerotic plaque rupture is one of the leading causes of stroke. Treatments for atherosclerosis can induce tissue damage during the deployment of an intravascular device or through external tissue clamping during surgery. In this paper, a constituent specific study was performed to investigate the role of the ground matrix and collagen fibres of arterial tissue in response to supra-physiological loads. Cyclic mechanical tests were conducted on intact and collagenase-digested strips of porcine common carotid arteries. Using these tests, four passive damage-relevant phenomena were studied, namely (i) Mullins effect, (ii) hysteresis, (iii) permanent set and (iv) matrix failure and fibre rupture. A constitutive model was also developed to capture all of these damage-relevant phenomena using a continuum damage mechanics approach. The implemented constitutive model was fit to experimental results for both intact and digested samples. The results of this work demonstrate the important role of the ground matrix in the permanent deformation of the arterial tissue under high loads. Supra-physiological load-induced tissue damage may play a key role in vascular remodelling in arteries with atherosclerosis or following interventional procedures.     

Identification of an intracellular domain of the EGF receptor required for high-affinity binding of EGF

Although all EGF receptors in EGF receptor-expressing cells are molecularly identical, they can be subdivided in two different classes that have either a high or a low affinity for EGF. Specifically the high-affinity class is associated with filamentous actin. To determine whether the interaction of the EGF receptor with actin induces its high-affinity state, we studied EGF-binding properties of an EGF receptor mutant that lacks the actin-binding site. Interestingly, we found that cells expressing this mutant receptor still display both high- and low-affinity classes of EGF receptors, indicating that the actin-binding domain does not determine the high-affinity binding state. By further mutational analysis we identified a receptor domain, within the tyrosine kinase domain, that regulates the affinity for EGF.     

Unprecedented Melioidosis Cases in Northern Australia Caused by an Asian Burkholderia pseudomallei Strain Identified by Using Large-Scale Comparative Genomics

Melioidosis is a disease of humans and animals that is caused by the saprophytic bacterium Burkholderia pseudomallei. Once thought to be confined to certain locations, the known presence of B. pseudomallei is expanding as more regions of endemicity are uncovered. There is no vaccine for melioidosis, and even with antibiotic administration, the mortality rate is as high as 40% in some regions that are endemic for the infection. Despite high levels of recombination, phylogenetic reconstruction of B. pseudomallei populations using whole-genome sequencing (WGS) has revealed surprisingly robust biogeographic separation between isolates from Australia and Asia. To date, there have been no confirmed autochthonous melioidosis cases in Australia caused by an Asian isolate; likewise, no autochthonous cases in Asia have been identified as Australian in origin. Here, we used comparative genomic analysis of 455 B. pseudomallei genomes to confirm the unprecedented presence of an Asian clone, sequence type 562 (ST-562), in Darwin, northern Australia. First observed in Darwin in 2005, the incidence of melioidosis cases attributable to ST-562 infection has steadily risen, and it is now a common strain in Darwin. Intriguingly, the Australian ST-562 appears to be geographically restricted to a single locale and is genetically less diverse than other common STs from this region, indicating a recent introduction of this clone into northern Australia. Detailed genomic and epidemiological investigations of new clinical and environmental B. pseudomallei isolates in the Darwin region and ST-562 isolates from Asia will be critical for understanding the origin, distribution, and dissemination of this emerging clone in northern Australia.