Metal cations modulate the bacteriochlorophyll–protein interaction in the light-harvesting 1 core complex from Thermochromatium tepidum

The light-harvesting 1 reaction center (LH1-RC) complex from Thermochromatium (Tch.) tepidum exhibits unusual Q_y absorption by LH1 bacteriochlorophyll-a (BChl-a) molecules at 915 nm, and the transition energy is finely modulated by the binding of metal cations to the LH1 polypeptides. Here, we demonstrate the metal-dependent interactions between BChl-a and the polypeptides within the intact LH1-RC complexes by near-infrared Raman spectroscopy. The wild-type LH1-RC (B915) exhibited Raman bands for the C3-acetyl and C13-keto CO stretching modes at 1637 and 1675 cm^? 1, respectively. The corresponding bands appeared at 1643 and 1673 cm^? 1 when Ca^2 + was biosynthetically replaced with Sr^2 + (B888) or at 1647 and 1669 cm^? 1 in the mesophilic counterpart, Allochromatium vinosum. These results indicate the significant difference in the BChl–polypeptide interactions between B915 and B888 and between B915 and the mesophilic counterpart. The removal of the original metal cations from B915 and B888 resulted in marked band shifts of the C3-acetyl/C13-carbonyl νCO modes to ~ 1645/~ 1670 cm^? 1, supporting a model in which the metal cations are involved in the fine-tuning of the hydrogen bonding between the BChl-a and LH1-polypeptides. Interestingly, the interaction modes were almost identical between the Ca^2 +-depleted B915 and Sr^2 +-depleted B888 and between B915 and Ca^2 +-substituted B888, despite the significant differences in their LH1 Q_y peak positions and the denaturing temperatures, as revealed by differential scanning calorimetry. These results suggest that not only the BChl–polypeptide interactions but some structural origin may be involved in the unusual Q_y red-shift and the enhanced thermal stability of the LH1-RC complexes from Tch. tepidum.     

Intestinal perfusion indicates high reliance on paracellular nutrient absorption in an insectivorous bat Tadarida brasiliensis

Flying vertebrates have been hypothesized to have a high capacity for paracellular absorption of nutrients. This could be due to high permeability of the intestines to nutrient-sized molecules (i.e., in the size range of amino acids and glucose, MW 75–180 Da). We performed intestinal luminal perfusions of an insectivorous bat, Tadarida brasiliensis. Using radio-labeled molecules, we measured the uptake of two nutrients absorbed by paracellular and transporter-mediated mechanisms (l-proline, MW 115 Da, and d-glucose, MW 180 Da) and two carbohydrates that have no mediated transport (l-arabinose, MW 150 Da, and lactulose, MW 342 Da). Absorption of lactulose (0.61 ± 0.06 nmol min^? 1 cm^? 1) was significantly lower than that of the smaller arabinose (1.09 ± 0.04 nmol min^? 1 cm^? 1). Glucose absorption was significantly lower than that of proline at both nutrient concentrations (10 mM and 75 mM). Using the absorption of arabinose to estimate the portion of proline absorption that is paracellular, we calculated that 25.1 ± 3.0% to 66.2 ± 7.8% of proline absorption is not transporter-mediated (varying proline from 1 mM to 75 mM). These results confirm our predictions that 1) paracellular absorption is molecule size selective, 2) absorption of proline would be greater than glucose absorption in an insectivore, and 3) paracellular absorption represents a large fraction of total nutrient absorption in bats.     

Photobiomodulation of human adipose-derived stem cells using 810 nm and 980 nm lasers operates via different mechanisms of action

Photobiomodulation (PBM) using red or near-infrared (NIR) light has been used to stimulate the proliferation and differentiation of adipose-derived stem cells. The use of NIR wavelengths such as 810 nm is reasonably well accepted to stimulate mitochondrial activity and ATP production via absorption of photons by cytochrome c oxidase. However, the mechanism of action of 980 nm is less well understood. Here we study the effects of both wavelengths (810 nm and 980 nm) on adipose-derived stem cells in vitro. Both wavelengths showed a biphasic dose response, but 810 nm had a peak dose response at 3 J/cm² for stimulation of proliferation at 24 h, while the peak dose for 980 nm was 10–100 times lower at 0.03 or 0.3 J/cm². Moreover, 980 nm (but not 810 nm) increased cytosolic calcium while decreasing mitochondrial calcium. The effects of 980 nm could be blocked by calcium channel blockers (capsazepine for TRPV1 and SKF96365 for TRPC channels), which had no effect on 810 nm. To test the hypothesis that the chromophore for 980 nm was intracellular water, which could possibly form a microscopic temperature gradient upon laser irradiation, we added cold medium (4 °C) during the light exposure, or pre-incubated the cells at 42 °C, both of which abrogated the effect of 980 nm but not 810 nm. We conclude that 980 nm affects temperature-gated calcium ion channels, while 810 nm largely affects mitochondrial cytochrome c oxidase.     

The effects of iodine on kidney bean starch: Films and pasting properties

In the present study the effect of iodine on the structural characteristics (by infrared spectroscopy and X-ray) of films made from kidney bean starch was evaluated. The pasting properties as affected by iodine and glycerol were also evaluated. Kidney bean starch showed C-type (mixture of A- and B-type) crystalline structure, the conversion of starch into films resulted into reduction in intensity of diffractograms. The starch powder FTIR spectra had peaks centered at 1020 and 995 cm^?1 with a higher intensity at 1020 cm^?1, which is consistent with a partially crystalline material since fully crystalline material show similar intensity peaks centered around 1020 and 1006 cm^?1. Films without iodine showed one main peak centered around 1000 cm^?1 consistent with a disordered state similar to that in gelatinised starch. Iodine addition gradually increased the intensity of the bands around 1020 cm^?1 consistent with the formation of more ordered conformation similar to that in the crystalline material. Iodine encourages the formation of helical structures, however, the formation of crystalline material cannot be inferred. The increasing amounts of iodine up to 0.33% level progressively increased the peak-, through- and breakdown-viscosity. Iodine beyond 0.33% level gradually decreased peak-, trough-, breakdown- and setback-viscosity. Pasting temperature gradually increased with the increase in iodine.     

A Kazal-type serine proteinase inhibitor from chicken liver (clTI-1): Purification,primary structure,and inhibitory properties

Low-molecular-mass trypsin inhibitor (clTI-1; chicken liver Trypsin Inhibitor-1) was purified from chicken liver by extraction with perchloric acid, ammonium sulfate precipitation, a combination of ethanol-acetone fractionation followed by gel filtration, ion-exchange chromatography and RP-HPLC on a C18 column. The inhibitor occurs in two isoforms with molecular masses of 5938.56 and 6026.29 Da (determined by MALDI TOFF mass spectrometry). The complete amino acid sequences of both isoforms were determined (UniProtKB/Swiss-Prot P85000; ISK1L_CHICK). The inhibitor shows a high homology to Kazal-type family inhibitors, especially to trypsin/acrosin inhibitors and pancreatic secretory trypsin inhibitors. clTI-1 inhibits both bovine and porcine trypsin (K_a = 1.1 × 10⁹ M^?1 and 2.5 × 10⁹ M^?1, respectively). Significant differences were shown in the inhibition of the anionic and cationic forms of chicken trypsin (K_a = 4.5 × 10⁸ M^?1 and 1.2 × 10¹⁰ M^?1). Weak interaction with human plasmin (K_a = 1.2 × 10⁷ M^?1) was also revealed.     

Structural changes that occur upon photolysis of the Fe(II)a3–CO complex in the cytochrome ba3-oxidase of Thermus thermophilus: A combined X-ray crystallographic and infrared spectral study demonstrates CO binding to CuB

The purpose of the work was to provide a crystallographic demonstration of the venerable idea that CO photolyzed from ferrous heme-a₃ moves to the nearby cuprous ion in the cytochrome c oxidases. Crystal structures of CO-bound cytochrome ba₃-oxidase from Thermus thermophilus, determined at ~ 2.8–3.2 Å resolution, reveal a Fe–C distance of ~ 2.0 Å, a Cu–O distance of 2.4 Å and a Fe–C–O angle of ~ 126°. Upon photodissociation at 100 K, X-ray structures indicate loss of Fe_a3–CO and appearance of Cu_B–CO having a Cu–C distance of ~ 1.9 Å and an O–Fe distance of ~ 2.3 Å. Absolute FTIR spectra recorded from single crystals of reduced ba₃–CO that had not been exposed to X-ray radiation, showed several peaks around 1975 cm^? 1; after photolysis at 100 K, the absolute FTIR spectra also showed a significant peak at 2050 cm^? 1. Analysis of the ‘light’ minus ‘dark’ difference spectra showed four very sharp CO stretching bands at 1970 cm^? 1, 1977 cm^? 1, 1981 cm^? 1, and 1985 cm^? 1, previously assigned to the Fe_a3–CO complex, and a significantly broader CO stretching band centered at ~ 2050 cm^? 1, previously assigned to the CO stretching frequency of Cu_B bound CO. As expected for light propagating along the tetragonal axis of the P4₃2₁2 space group, the single crystal spectra exhibit negligible dichroism. Absolute FTIR spectrometry of a CO-laden ba₃ crystal, exposed to an amount of X-ray radiation required to obtain structural data sets before FTIR characterization, showed a significant signal due to photogenerated CO₂ at 2337 cm^? 1 and one from traces of CO at 2133 cm^? 1; while bands associated with CO bound to either Fe_a3 or to Cu_B in “light” minus “dark” FTIR difference spectra shifted and broadened in response to X-ray exposure. In spite of considerable radiation damage to the crystals, both X-ray analysis at 2.8 and 3.2 Å and FTIR spectra support the long-held position that photolysis of Fe_a3–CO in cytochrome c oxidases leads to significant trapping of the CO on the Cu_B atom; Fe_a3 and Cu_B ligation, at the resolutions reported here, are otherwise unaltered. This article is part of a Special Issue entitled: Respiratory Oxidases.     

Colonization dynamics of biofilm-associated ciliate morphotypes at different flow velocities

The impact of flow velocity on initial ciliate colonization dynamics on surfaces were studied in the third order Ilm stream (Thuringia, Germany) at a slow flowing site (0.09 m s^?1) and two faster flowing sites (0.31 m s^?1) and in flow channels at 0.05, 0.4, and 0.8 m s^?1. At the slow flowing stream site, surfaces were rapidly colonized by ciliates with up to 60 cells cm^?2 after 24 h. In flow channels, the majority of suspended ciliates and inorganic matter accumulated at the surface within 4.5 h at 0.05 m s^?1. At 0.4 m s^?1 the increase in ciliate abundance in the biofilm was highest between 72 and 168 h at about 3 cells cm^?2 h^?1. Faster flow velocities were tolerated by vagile flattened ciliates that live in close contact to the surface. Vagile flattened and round filter feeders preferred biofilms at slow flow velocities. Addition of inorganic particles (0, 0.6, and 7.3 mg cm^?2) did not affect ciliate abundance in flow channel biofilms, but small ciliate species dominated and number of species was lowest (16 species cm^?2) in biofilms at high sediment content. Although different morphotypes dominated the communities at contrasting flow velocities, all functional groups contributed to initial biofilm communities implementing all trophic links within the microbial loop.     

Optimization of intravascular shear stress assessment in vivo

The advent of microelectromechanical systems (MEMS) sensors has enabled real-time wall shear stress (WSS) measurements with high spatial and temporal resolution in a 3-D bifurcation model. To optimize intravascular shear stress assessment, we evaluated the feasibility of catheter/coaxial wire-based MEMS sensors in the abdominal aorta of the New Zealand white (NZW) rabbits. Theoretical and computational fluid dynamics (CFD) analyses were performed. Fluoroscope and angiogram provided the geometry of aorta, and the Doppler ultrasound system provided the pulsatile flow velocity for the boundary conditions. The physical parameters governing the shear stress assessment in NZW rabbits included (1) the position and distance from which the MEMS sensors were mounted to the terminal end of coaxial wire or the entrance length, (L_e), (2) diameter ratios of aorta to the coaxial wire (D_aorta /D_{coaxial wire}=1.5–9.5), and (3) the range of Reynolds numbers (116–1550). At an aortic diameter of 2.4 mm and a maximum Reynolds number of 212 (a mean Reynolds number of 64.2), the time-averaged shear stress (τ_ave) was computed to be 10.06 dyn cm^?2 with a systolic peak at 33.18 dyn cm^?2. In the presence of a coaxial wire (D_aorta /D_{coaxial wire}=6 and L_e=1.18 cm), the τ_ave value increased to 15.54 dyn cm^?2 with a systolic peak at 51.25 dyn cm^?2. Real-time intravascular shear stress assessment by the MEMS sensor revealed an τ_ave value of 11.92 dyn cm^?2 with a systolic peak at 47.04 dyn cm^?2. The difference between CFD and experimental τ_ave was 18.5%. These findings provided important insights into packaging the MEMS sensors to optimize in vivo shear stress assessment.     

Iron containing keratinolytic metallo-protease produced by Chryseobacterium gleum

Chryseobacterium gleum exhibited complete dissolution of whole chicken-feathers (10 g l^?1, pH 8) after 72 h at 30 °C through synthesis of keratinolytic protease when inoculated at 1% (v/v). This enzyme was purified to 67-fold with yield of 2.25% having a specific activity of 1670 U mg^?1 and ～36 kDa Mw. MALDI-TOF MS of this keratinase showed some similarity with the keratinase peptides of Bacillus subtilis (BOFXJ2). The keratinase action was inhibited by EDTA, iodoacetamide and metal ions like mercury, copper and zinc (1 mM each), while it was enhanced by iron and calcium. Keratinase showed presence of 3 mM of Fe M^?1 as tested by atomic absorption spectroscopy and addition of Fe in its apoenzyme retained about 79% of original residual feather degradation activity which portrayed it to be metalloprotease. Purified keratinase revealed significant degradation (85%) of feather concentrate (20 g l^?1) to 3.9 μM ml^?1 of free amino groups in 24 h at an initial pH of 8.0, 30 °C and 120 rpm shaking. This keratinase activity can be controlled precisely by presence of chemical or metal ions which could be of use in biotechnology industry while the culture can be used in poultry waste management.     

Inhibition of acetylcholinesterase by chromophore-linked fluorophosphonates

Fluorophosphonate (FP) head groups were tethered to a variety of chromophores (C) via a triazole group and tested as FPC inhibitors of recombinant mouse (rMoAChE) and electric eel (EEAChE) acetylcholinesterase. The inhibitors showed bimolecular inhibition constants (k_i) ranging from 0.3 × 10⁵ M^?1 min^?1 to 10.4 × 10⁵ M^?1 min^?1. When tested against rMoAChE, the dansyl FPC was 12.5-fold more potent than the corresponding inhibitor bearing a Texas Red as chromophore, whereas the Lissamine and dabsyl chromophores led to better anti-EEAChE inhibitors. Most inhibitors were equal or better inhibitors of rMoAChE than EEAChE. 3-Azidopropyl fluorophosphonate, which served as one of the FP head groups, showed excellent inhibitory potency against both AChE’s (? 1 × 10⁷ M^?1 min^?1) indicating, in general, that addition of the chromophore reduced the overall anti-AChE activity. Covalent attachment of the dabsyl-FPC analog to rMoAChE was demonstrated using size exclusion chromatography and spectroscopic analysis, and visualized using molecular modeling.     

Characterization of lactase-conjugated magnetic nanoparticles

Conjugation of lactase to magnetic nanoparticles is of interest in biosensor and ingredient processing applications that require high enzyme concentration and catalyst separation from the reaction stream. However, little is known about the effects of these materials on the physicochemical attributes of conjugated lactase. Lactase (Aspergillus oryzae) was covalently attached by carbodiimide chemistry to carboxylic-acid functionalized magnetic particles having a hydrodynamic radius of 18 nm. The resulting enzyme–nanoparticle conjugates were characterized with regard to particle size, zeta potential, enzyme kinetics, temperature and pH stability, catalyst recovery, and secondary structure changes. Following attachment, the materials retained colloidal stability and individual particle characteristics with a zeta potential of ?33 mV compared to ?46 mV for the native particle. The conjugated enzyme showed no changes in secondary structure and exhibited significant catalytic activity with a catalytic efficiency of 2.8 × 10³ M^?1 s^?1 compared to 2.5 × 10³ M^?1 s^?1 for the native enzyme. Relative to the free enzyme, the conjugated enzyme was recovered for repeated use with 78% activity retained after five cycles. This work demonstrates that carboxylic-acid functionalized magnetic nanoparticles can be utilized as a means of producing a simple and effective conjugated-lactase system that achieves both particle and enzyme stability.     

Characterization of a novel laccase from the isolated Coltricia perennis and its application to detoxification of biomass

A highly efficient laccase-producing fungus was isolated from soil and identified as Coltricia perennis SKU0322 by its morphology and by comparison of its internal transcribed spacer (ITS) rDNA gene sequence. Extracellular laccase (Cplac) from C. perennis was purified to homogeneity by anion-exchange and gel filtration chromatography. Cplac is a monomeric glycoprotein with 12% carbohydrate content and a molecular mass of 66 kDa determined by polyacrylamide-gel electrophoresis. Ultraviolet-visible absorption spectroscopy observed type 1 and type 3 copper signals from Cplac. The enzyme acted optimally at pH 3–4 and 75 °C. Its optimal activity was with 2,2-azino-bis (3-ethylbenzothiazoline-6-sulfonate) (ABTS), it also oxidized various lignin-related phenols. The enzyme was characterized as a multi-copper blue laccase by its substrate specificity and internal amino acid sequence. It showed a higher catalytic efficiency towards ABTS (k_cat/K_m = 18.5 s^?1 μM^?1) and 2,6-dimethoxyphenol (k_cat/K_m = 13.9 s^?1 μM^?1) than any other reported laccase. Its high stability and catalytic efficiency suggest its suitability for industrial applications: it detoxified phenolic compounds in acid-pretreated rice straw and enhanced saccharification yield.     

1-Allyl-3-methylimidazolium chloride plasticized-corn starch as solid biopolymer electrolytes

Ionic liquids (ILs), 1-allyl-3-methylimidazolium chloride ([amim]Cl) is found to be a novel plasticizer for cornstarch. [Amim]Cl-plasticized starch film also has a potential application as solid biopolymer electrolytes. In this study, different proportional [amim]Cl/glycerol mixtures are used to plasticize starch by casting. Atomic force microscopy (AFM) finds the diameter of residual starch granules existed in [amim]Cl or glycerol-plasticized starch films is only about 10 nm. However, glycerol can form more intensive hydrogen bond with starch than [amim]Cl detected by Fourier transform infrared (FT-IR) spectroscopy. So some novel ILs with high concentration and active hydrogen bond acceptors are necessary. Moreover, high [amim]Cl content can improve the water absorption and conductance of TPS film simultaneously. The conductance of TPS film with 30 wt% [amim]Cl content can achieve to 10^?1.6 S cm^?1 at 14.5 wt% water content.     

Laccase immobilization on mesostructured cellular foams affords preparations with ultra high activity

Extracellular laccase produced by the wood-rotting fungus Cerrena unicolor was immobilized covalently on the mesostructured siliceous cellular foams (MCFs) functionalised using various organosilanes with amine and glycidyl groups. The experiments indicated that laccase bound via glutaraldehyde to MCFs modified using 2-aminoethyl-3-aminopropyltrimethoxysilane remains very active. In the best biocatalyst activity was about 42,700 U mL^?1 carrier (66,800 U mg^?1 bound protein), and hence significantly higher than ever reported before. Optimisation of the immobilization procedure with respect to protein concentration, pH of coupling mixture and the enzyme purity afforded the biocatalyst with activity of about 90,980 U mL^?1. For the best preparation, thermal- and pH-stability, and activity profiles were determined. Experiments carried out in a batch reactor showed that k_cat/K_m for immobilized enzyme (0.88 min^?1 μM^?1) was acceptable lower than the value obtained for the native enzyme (2.19 min^?1 μM^?1). Finally, potentials of the catalysts were tested in the decolourisation of indigo carmine without redox-mediators. Seven consecutive runs with the catalysts separated by microfiltration proved that adsorption of the dye onto the carrier and enzymatic oxidation contribute to the efficient decolourisation without loss of immobilized enzyme activity.     

Influence of urea and calcium dosage on the effectiveness of bacterially induced carbonate precipitation on limestone

Bacterially induced carbonate precipitation has been explored for the protection and consolidation of ornamental stone. Attempts to improve the efficiency of this biodeposition process were primarily focused on the microbial aspects, i.e. type of microorganism and metabolic pathway. In this study, the influence of the chemical parameters, i.e. concentration of calcium salts and urea, on the effectiveness of the biodeposition treatment has been examined. The amount of calcium carbonate that can be precipitated in the stone is conditioned both by the amount of cells retained in the stone and the concentration of urea and calcium used. From sonication experiments, a good consolidation was observed for limestone prisms treated with a calcium dosage of 17 g Ca²⁺ m^?2 with no improvement at higher concentrations. For limestone prisms of 4 cm × 2 cm × 1 cm, the biodeposition treatment resulted in a 63% lower weight loss upon sonication compared to untreated specimens. The waterproofing effect was observed to increase with increasing calcium dosages. While for a calcium dosage of 17 g Ca²⁺ m^?2 the water absorption was similar to that of untreated specimens, concentrations of 67 g Ca²⁺ m^?2 resulted in a 50% decrease of the rate of water absorption. For calcium dosages higher than 34 g Ca²⁺ m^?2 a significant change in the visual aspect (ΔE > 6) of the treated stones could be observed. Overall, the urea/calcium chloride-based biodeposition treatment attained a protective performance comparable with that of the commonly used ethylsilicates.     

Synthesis of α-tocohexaenol (α-T6) a fluorescent,oxidatively sensitive polyene analogue of α-tocopherol

A polyunsaturated analogue of α-tocopherol was synthesized that is both fluorescent and sensitive to peroxidative chemistry that occurs in phospholipid membranes. α-Tocohexaenol 1, [(S)-2,5,7,8-tetramethyl-2-((1E/Z,3E,5E,7E,9E)-4,8,12-trimethyltrideca-1,3,5,7,9,11-hexaenyl)chroman-6-ol, α-T6] was prepared by condensing a known triene fragment triphenyl-(2,6-dimethyl-octa-2,4,6-trienoic acid methyl ester)-phosphonium bromide with a protected chromanol aldehyde, (2S)-6-{[tert-butyl(dimethyl)silyl]oxy}-2,5,7,8-tetra-methyl-3,4-dihydro-2H-chromene-2-carbaldehyde. The full side chain was then completed with isopentyl(tri-n-butyl)phosphonium bromide to give 1. The geometry of the C1′?C2′ alkene appears to be Z (cis) although the coupling constants of the olefinic protons are intermediate between values normally assigned to E and Z-isomers. In ethanol, α-T6 has a maximum absorption at 368 nm with an absorption coefficient of 45,000 M^?1 cm^?1, and displays a maximum fluorescence emission at 523 nm. The susceptibility of α-T6 to peroxidative chemistry was dependent on the concentration of azo-initiators of lipid oxidation in acetonitrile solution as well as in phospholipid vesicles. A loss of fluorescence at 520 nm was observed when α-T6 (vesicles or α-T6-lipid mixtures) was exposed to peroxidative conditions, and this loss mirrored the production of conjugated dienes and trienes during the peroxidation of bulk phospholipids. Addition of natural α-tocopherol during the AMVN induced oxidation of 4 μM α-T6 and 0.5 mg/ml soybean PC induced a characteristic lag phase, after which the fluorescence of α-T6 began to lessen. Thus, α-T6 may be a useful reporter not only of tocopherol location in cells, but also of the extent of peroxidative events.     

Detection and quantification of 4-substituted phenols: a comparison of mushroom tyrosinase and cell extracts of Pseudomonas putida F6

A comparison of the spectrophotometric detection and quantification of a number of 4-substituted phenols by two sources of the enzyme tyrosinase (Agaricus bisporus (mushroom) versus Pseudomonas putida) is described. Incubation of either source of tyrosinase with selected 4-substituted phenols results in the formation of coloured products that absorb light maximally within a narrow wavelength range (400–423 nm). The inclusion of the nucleophile 3-methyl-2-benzothiazolinone (MBTH) in the tyrosinase assay results in more intensely coloured products that also absorb light within a narrow wavelength range (440–475 nm). The molar extinction coefficient of the reaction products in the tyrosinase and tyrosinase–MBTH assay differed dramatically with values between 714–1580 and 14213–26563 M⁻¹ cm⁻¹, respectively. The addition of MBTH improved the sensitivity of the reaction between 1.3- and 100-fold, depending on the substrate and source of the enzyme. The limit of detection of 4-substituted phenols also varied according to substrate and the source of enzyme used in the assay. The lowest detectable concentration of 4-substituted phenol was 2.5 μM 4-hydroxyphenoxy acetic acid in the presence of mushroom tyrosinase and MBTH and 2.5 μM 2-(4-hydroxyphenyl) ethanol in the presence of cell extract of P. putida F6 and MBTH.     

Knee flexor strength and bicep femoris electromyographical activity is lower in previously strained hamstrings

The aim of this study was to determine if athletes with a history of hamstring strain injury display lower levels of surface EMG (sEMG) activity and median power frequency in the previously injured hamstring muscle during maximal voluntary contractions. Recreational athletes were recruited, 13 with a history of unilateral hamstring strain injury and 15 without prior injury. All athletes undertook isokinetic dynamometry testing of the knee flexors and sEMG assessment of the biceps femoris long head (BF) and medial hamstrings (MHs) during concentric and eccentric contractions at ±180 and ±60° s^?1. The knee flexors on the previously injured limb were weaker at all contraction speeds compared to the uninjured limb (+180° s^?1 p = 0.0036; +60° s^?1 p = 0.0013; ?60° s^?1 p = 0.0007; ?180° s^?1 p = 0.0007) whilst sEMG activity was only lower in the BF during eccentric contractions (?60° s^?1 p = 0.0025; ?180° s^?1 p = 0.0003). There were no between limb differences in MH sEMG activity or median power frequency from either BF or MH in the injured group. The uninjured group showed no between limb differences in any of the tested variables. Secondary analysis comparing the between limb difference in the injured and the uninjured groups, confirmed that previously injured hamstrings were mostly weaker (+180° s^?1 p = 0.2208; +60° s^?1 p = 0.0379; ?60° ^?1 p = 0.0312; ?180° s^?1 p = 0.0110) and that deficits in sEMG were confined to the BF during eccentric contractions (?60° s^?1 p = 0.0542; ?180° s^?1 p = 0.0473). Previously injured hamstrings were weaker and BF sEMG activity was lower than the contralateral uninjured hamstring. This has implications for hamstring strain injury prevention and rehabilitation which should consider altered neural function following hamstring strain injury.     

Syntheses,structures, and magnetic properties of heterobimetallic complexes based on tetracyanometallic building blocks

Two tetracyanometalate building blocks, [Fe(5,5′-dmbipy)(CN)₄]^? (2) and [Fe(4,4′-dmbipy)(CN)₄]^? (3) (5,5′-dmbipy = 5,5′-dimethyl-2,2′-bipyridine; 4,4′-dmbipy = 4,4′-dimethyl-2,2′-bipyridine), and two cyano-bridged heterobimetallic complexes, [Cu₂(bpca)₂(H₂O)₂Fe₂(5,5′-dmbipy)₂(CN)₈] · 2[Cu(bpca)Fe(5,5′-dmbipy)(CN)₄] · 4H₂O (4) and [Cu(bpca)Fe(4,4′-dmbipy)(CN)₄]_n (5) (bpca = bis(2-pyridylcarbonyl)amidate), have been synthesized and structurally characterized. Complex 4 contains two dinuclear and one tetranuclear heterobimetallic clusters in an asymmetric unit whereas the structure of complex 5 features a one-dimensional heterobimetallic zigzag chain. The Cu(II) ion is penta-coordinated in the form of a distorted square-based pyramid. Magnetic studies show ferromagnetic coupling between Cu(II) and Fe(III) ions with g = 2.28, J₁ = 2.64 cm^?1, J₂ = 5.40 cm^?1 and TIP = ?2.36 × 10^?3 for complex 4, and g = 2.17, J = 4.82 cm^?1 and zJ′ = 0.029 cm^?1 for complex 5.     

Cloning,overexpression, purification,and site-directed mutagenesis of xylitol-2-dehydrogenase from Candida albicans

Xylitol-2-dehydrogenase from Candida albicans was cloned and overexpressed in Escherichia coli. The purified recombinant XDH has an apparent molecular weight of 40 kDa which belongs to the medium chain alcohol dehydrogenase family and exclusively uses NAD⁺ as a cofactor. The recombinant caXDH has a K_M of 8.8 mM and 37.7 μM using the substrate xylitol and NAD⁺, respectively, and its catalytic efficiency is 53,200 min^?1 mM^?1. Following site-directed mutagenesis, one of the engineered caXDHs with six mutations at Ser95Cys, Ser98Cys, Tyr101Cys, Asp206Ala, Ile207Arg, and Phe208Ser shifted its cofactor dependence from NAD⁺ to NADP⁺ in which the K_M and k_cat/K_M towards NADP⁺ are 119 μM and 26,200 min^?1 mM^?1, respectively.