Loop grafting of Bacillus subtilis lipase A: inversion of enantioselectivity

Lipases are successfully applied in enantioselective biocatalysis. Most lipases contain a lid domain controlling access to the active site, but Bacillus subtilis Lipase A (LipA) is a notable exception: its active site is solvent exposed. To improve the enantioselectivity of LipA in the kinetic resolution of 1,2-O-isopropylidene-sn-glycerol (IPG) esters, we replaced a loop near the active-site entrance by longer loops originating from Fusarium solani cutinase and Penicillium purpurogenum acetylxylan esterase, thereby aiming to increase the interaction surface for the substrate. The resulting loop hybrids showed enantioselectivities inverted toward the desired enantiomer of IPG. The acetylxylan esterase-derived variant showed an inversion in enantiomeric excess (ee) from -12.9% to +6.0%, whereas the cutinase-derived variant was improved to an ee of +26.5%. The enantioselectivity of the cutinase-derived variant was further improved by directed evolution to an ee of +57.4%.     

BOLD MRI response to hypercapnic hyperoxia in patients with meningiomas: correlation with Gadolinium-DTPA uptake rate

Because meningiomas tend to recur after (partial) surgical resection, radiotherapy is increasingly being applied for the treatment of these tumors. Radiation dose levels are limited, however, to avoid radiation damage to the surrounding normal tissue. The radiosensitivity of tumors can be improved by increasing tumor oxygen levels. The aim of this study was to investigate if breathing a hyperoxic hypercapnic gas mixture could improve the oxygenation of meningiomas. Blood oxygen level-dependent magnetic resonance imaging and dynamic Gadolinium (Gd)-DTPA contrast-enhanced MRI were used to assess changes in tumor blood oxygenation and vascularity, respectively. Ten meningioma patients were each studied twice; without and with breathing a gas mixture consisting of 2% CO(2) and 98% O(2). Values of T(2)* and the Gd-DTPA uptake rate k(ep) were calculated under both conditions. In six tumors a significant increase in the value of T(2)* in the tumor was found, suggesting an improved tumor blood oxygenation, which exceeded the effect in normal brain tissue. Contrarily, two tumors showed a significant T(2)* decrease. The change in T(2)* was found to correlate with both k(ep) and with the change in k(ep). The presence of both vascular effects and oxygenation effects and the heterogeneous response to hypercapnic hyperoxia necessitates individual assessment of the effects of breathing a hyperoxic hypercapnic gas mixture on meningiomas. Thus, the current MRI protocol may assist in radiation treatment selection for patients with meningiomas.     

Determination of nitration degrees for the birch pollen allergen Bet v 1

Nitration of tyrosine residues in the major birch pollen allergen Bet v 1 may alter the allergenic potential of the protein. The kinetics and mechanism of the nitration reaction, however, have not yet been well characterized. To facilitate further investigations, an efficient method to quantify the nitration degree (ND) of small samples of Bet v 1 is required. Here, we present a suitable method of high-performance liquid chromatography coupled to a diode array detector (HPLC-DAD) that can be photometrically calibrated using the amino acids tyrosine (Tyr) and nitrotyrosine (NTyr) without the need for nitrated protein standards. The new method is efficient and in agreement with alternative methods based on hydrolysis and amino acid analysis of tetranitromethane (TNM)-nitrated Bet v 1 standards as well as samples from nitration experiments with peroxynitrite. The results confirm the applicability of the new method for the investigation of the reaction kinetics and mechanism of protein nitration.

Antimicrobial Activity and Wound-Healing Capacity of Birch,Beech and Larch Bark Extracts

Bark is a major by-product of woodworking industries. The contents of several wood species are known to harbor antimicrobial, antiviral, anti-inflammatory and wound-healing capacities. The aim of this work was to identify beneficial properties of Austrian larch, birch and beech bark extracts for their potential usage as additives or active ingredients in dermatological applications. Bacterial agar diffusion assay and resazurin-based broth microdilution assay were used to evaluate anti-bacterial activity. To gain more insight into the cellular response to bark extracts, viability-, scratch-assays and ELISAs were performed. Birch and beech extracts showed strong antimicrobial activities against Gram-positive bacteria, including Cutibacterium acnes, Staphylococcus epidermidis and MRSA. Wound closure was enhanced with birch and beech extracts as compared to controls in the scratch-assays. Whereas beneficial properties of birch bark components have previously been described, the similar effects of beech extracts are novel. The combined positive effect on wound-healing and antimicrobial activity has great potential for the treatment of various skin diseases, including acne in future dermal applications.     

Producing translationally cold, ground-state CO molecules

Carbon monoxide molecules in their electronic, vibrational, and rotational ground state are highly attractive for trapping experiments. The optical or ac electric traps that can be envisioned for these molecules will be very shallow, however, with depths in the sub-milliKelvin range. Here, we outline that the required samples of translationally cold CO (X(1)Σ(+), v' = 0, N' = 0) molecules can be produced after Stark deceleration of a beam of laser-prepared metastable CO (a(3)Π(1)) molecules followed by optical transfer of the metastable species to the ground state via perturbed levels in the A(1)Π state. The optical transfer scheme is experimentally demonstrated and the radiative lifetimes and the electric dipole moments of the intermediate levels are determined.     

Thermodynamic Uncertainty Relations

Bohr and Heisenberg suggested that the thermodynamical quantities of temperature and energy are complementary in the same way as position and momentum in quantum mechanics. Roughly speaking their idea was that a definite temperature can be attributed to a system only if it is submerged in a heat bath, in which case energy fluctuations are unavoidable. On the other hand, a definite energy can be assigned only to systems in thermal isolation, thus excluding the simultaneous determination of its temperature. Rosenfeld extended this analogy with quantum mechanics and obtained a quantitative uncertainty relation in the form U (1/T) k, where k is Boltzmann's constant. The two extreme cases of this relation would then characterize this complementarity between isolation (U definite) and contact with a heat bath (T definite). Other formulations of the thermodynamical uncertainty relations were proposed by Mandelbrot (1956, 1989), Lindhard (1986), and Lavenda (1987, 1991). This work, however, has not led to a consensus in the literature. It is shown here that the uncertainty relation for temperature and energy in the version of Mandelbrot is indeed exactly analogous to modern formulations of the quantum mechanical uncertainty relations. However, his relation holds only for the canonical distribution, describing a system in contact with a heat bath. There is, therefore, no complementarily between this situation and a thermally isolated system.     

Spatially Controlled Out‐of‐Equilibrium Host–Guest System under Electrochemical Control

Self‐assembly to create molecular and nanostructures is typically performed at the thermodynamic minimum. To achieve dynamic functionalities, such as adaptability, internal feedback, and self‐replication, there is a growing focus on out‐of‐equilibrium systems. This report presents the dynamic self‐assembly of an artificial host–guest system at an interface, under control by a dissipative electrochemical process using (electrical) energy, resulting in an out‐of‐equilibrium system exhibiting a supramolecular surface gradient. The gradient, its steepness, rate of formation, and complex surface composition after backfilling, as well as the surface compositions after switching between the different states of the system, are assessed and supported by modelling. Our method shows for the first time an artificial surface‐confined out‐of‐equilibrium system. The electrochemical process parameters provide not only control over the system in time, but also in space.