Isolation and characterization of a novel 5-oxoprolinase (without ATP-hydrolyzing) from Alcaligenes faecalis N-38A

A screening test was undertaken to isolate a microorganism that produced 5-oxoprolinase (without ATP-hydrolyzing). The 5-oxoprolinase (without ATP-hydrolyzing) activity (decyclization activity toward L-pyroglutamate) was found in a cell-free extract of Alcaligenes faecalis N-38A, newly isolated from a soil sample. The enzyme was purified as a homogeneous preparation. The molecular weight of the enzyme was estimated to be 47,000. The decyclization activity was specific for L-pyroglutamate, and independent of ATP and metal ions. The reaction was a reversible one, i.e., cyclization reaction of L-glutamate to yield pyroglutamate was identified.     

Clonal composition of malignant fibrous histiocytoma: analysis by PCR-based assay of the human androgen receptor gene (HUMARA)

Poly(A)+ mRNA was isolated from leaves of potato plants (Solanum tuberosum L. cv. Desiree) according to standard protocols. This poly(A)+ mRNA was injected via glass microcapillaries into oocytes that were surgically removed from the African clawed toad Xenopus laevis. As a control, oocytes were either injected with H2O or remained untreated. Three days after injection the oocytes were analyzed by two electrode voltage clamping. Current voltage analysis revealed that a K+ channel from potato was functionally expressed in injected oocytes. The identity of this K+ channel was confirmed by its substrate specificity and a shift in the reversal potential. In particular, when the outside K+ concentration was increased the reversal potential of poly(A)+ injected oocytes shifted to more positive values. Furthermore, K+ outward currents declined when the outside K+ concentration was raised from 0.1 to 100 mM. Inward currents increased with an elevation of the K+ concentration. Several pharmaceuticals were tested for their potential to block this K+ channel. As a result, the channel was completely blocked by BaCl2. A three state reaction kinetic model was used to simulate the currents through the K+ transport protein as function of the extracellular K+ concentration. In particular, the simulation revealed current voltage relations that exactly matched the measured ones. Saturation of current voltage curves emerged from the simulation as a consequence of high extracellular potassium concentration.     

Modeling and control of a finger-like mechanism using bending shape memory alloys

Microsystem Technologies - In this research a biologically inspired finger-like mechanism similar to human musculoskeletal system is developed based on Shape Memory Alloys (SMAs). SMA actuators are...     

Some new volatile compounds from the non-enzymic browning reaction of glucose-glutamic acid system

Summary The volatile compounds from the Maillard reaction in the glucose - glutamic acid system were analysed by combined gas chromatography - mass spectrometry. Of the 50 peaks detected, 42 components comprising 2 acids, 2 alcohols, 5 carbonyls, 2 esters, 4 lactones, 17 furans, 4 pyrazines and 3 pyrroles were identified, four of which tentatively. Decadien 2,4-al, 2-n-pentyl furoate, 2-(2-fury()-pyrazine. -nonalactone, 2-(2-fury()-5- or 6-methylpyrazine were previously not reported to occur in heated sugar - amino acid browning systems. The new compound, 5-formyloxymethyl-2-furfural has apparently not yet been recorded in the chemical literature.The compound 5-acetoxymethyl-2-furfural had an aroma close to that of bread whereas 5-formyloxymethyl-2-furfural exhibited a very faint sweet burnt aroma.

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Einige neue flüchtige Verbindungen aus der nicht-enzymatischen Bräunungsreaktion des Glucose-Glutaminsäure-Systems

Zusammenfassung Die flüchtigen Verbindungen aus der Maillardreaktion des Glucose-Glutaminsäure-Systems wurde mit Hilfe der Gaschromatographie/Massenspektrometrie analysiert. Von den 50 nachgewiesenen Peaks wurden 42 identifiziert: 2 Säuren, 2 Alkohole, 5 Carbonyle, 2 Ester, 4 Lactone, 17 Furane, 4 Pyrazine, 3 Pyrrole und 4 weitere provisorisch. Deca-2,4-dienal, 2-n-Pentylfluorat, 2-(2-Furyl)-pyrazin, -Nonalacton, 2-(2-Furyl)-5 oder -6-Methylpyrazin wurden bisher nicht in dem erhitzten Zucker-Aminosäuren-Bräunungssystem registriert. Die neue Verbindung 5-Formyloxymethyl-2-furfural ist anscheinend noch nicht in der chemischen Literatur festgestellt worden. Die Verbindung 5-Acetoxymethyl-2-furfural hat an ein Brot erinnerndes Aroma, während die 5-Formyloxymethyl-2-furfural-Verbindung ein sehr schwaches süßlich-verbranntes Aroma aufweist.

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Taken in part from the Ph. D. thesis of the Senior Author     

Annexin 1 Is a Component of eATP-Induced Cytosolic Calcium Elevation in Arabidopsis thaliana Roots

Extracellular ATP (eATP) has long been established in animals as an important signalling molecule but this is less understood in plants. The identification of Arabidopsis thaliana DORN1 (Does Not Respond to Nucleotides) as the first plant eATP receptor has shown that it is fundamental to the elevation of cytosolic free Ca²⁺ ([Ca²⁺]_cyt) as a possible second messenger. eATP causes other downstream responses such as increase in reactive oxygen species (ROS) and nitric oxide, plus changes in gene expression. The plasma membrane Ca²⁺ influx channels involved in eATP-induced [Ca²⁺]_cyt increase remain unknown at the genetic level. Arabidopsis thaliana Annexin 1 has been found to mediate ROS-activated Ca²⁺ influx in root epidermis, consistent with its operating as a transport pathway. In this study, the loss of function Annexin 1 mutant was found to have impaired [Ca²⁺]_cyt elevation in roots in response to eATP or eADP. Additionally, this annexin was implicated in modulating eATP-induced intracellular ROS accumulation in roots as well as expression of eATP-responsive genes.     

Molecular Docking Studies and Biological Evaluation of Berberine–Benzothiazole Derivatives as an Anti-Influenza Agent via Blocking of Neuraminidase

In this study, we have introduced newly synthesized substituted benzothiazole based berberine derivatives that have been analyzed for their in vitro and in silico biological properties. The activity towards various kinds of influenza virus strains by employing the cytopathic effect (CPE) and sulforhodamine B (SRB) assay. Several berberine–benzothiazole derivatives (BBDs), such as BBD1, BBD3, BBD4, BBD5, BBD7, and BBD11, demonstrated interesting anti-influenza virus activity on influenza A viruses (A/PR/8/34, A/Vic/3/75) and influenza B viral (B/Lee/40, and B/Maryland/1/59) strain, respectively. Furthermore, by testing neuraminidase activity (NA) with the neuraminidase assay kit, it was identified that BBD7 has potent neuraminidase activity. The molecular docking analysis further suggests that the BBD1–BBD14 compounds’ antiviral activity may be because of interaction with residues of NA, and the same as in oseltamivir.     

Modulation of Tubular pH by Acetazolamide in a Ca2+ Transport Deficient Mice Facilitates Calcium Nephrolithiasis

Proximal tubular (PT) acidosis, which alkalinizes the urinary filtrate, together with Ca²⁺ supersaturation in PT can induce luminal calcium phosphate (CaP) crystal formation. While such CaP crystals are known to act as a nidus for CaP/calcium oxalate (CaOx) mixed stone formation, the regulation of PT luminal Ca²⁺ concentration ([Ca²⁺]) under elevated pH and/or high [Ca²⁺] conditions are unknown. Since we found that transient receptor potential canonical 3 (TRPC3) knockout (KO; -/-) mice could produce mild hypercalciuria with CaP urine crystals, we alkalinized the tubular pH in TRPC3-/- mice by oral acetazolamide (0.08%) to develop mixed urinary crystals akin to clinical signs of calcium nephrolithiasis (CaNL). Our ratiometric (λ340/380) intracellular [Ca²⁺] measurements reveal that such alkalization not only upsurges Ca²⁺ influx into PT cells, but the mode of Ca²⁺ entry switches from receptor-operated to store-operated pathway. Electrophysiological experiments show enhanced bicarbonate related current activity in treated PT cells which may determine the stone-forming phenotypes (CaP or CaP/CaOx). Moreover, such alkalization promotes reactive oxygen species generation, and upregulation of calcification, inflammation, fibrosis, and apoptosis in PT cells, which were exacerbated in absence of TRPC3. Altogether, the pH-induced alteration of the Ca²⁺ signaling signature in PT cells from TRPC3 ablated mice exacerbated the pathophysiology of mixed urinary stone formation, which may aid in uncovering the downstream mechanism of CaNL.     

Facile Deposition of Silver Nanoparticles on Photonic Cellulose Nanocrystals Films: A Study on Solvent Stability and Post Antibacterial Activity

Cellulose nanocrystals (CNCs) incorporated with silver nanoparticles (AgNPs) photonic films have drawn considerable attention due to their plasmonic chiroptical activity. However, the exploitation of some fundamental properties for practical use such as the affinity analysis of metal nanoparticles attached to the surface of photonic films according to the solvent compatibility and antibacterial activity under physical conditions has yet not been studied. Hence, a facile process of in situ deposition of AgNPs into the chiral structure of CNC films is proposed. CNC photonic films, cross-linked by glutaraldehyde are prepared. This interaction generated the solvents-stable photonic film with a considerable amount of unreacted aldehyde functional groups that facilitates the reduction of Ag salt to AgNPs. The formed AgNPs in the photonic films show excellent stability over immersion in various polar and non-polar solvents. The post-solvent treated photonic films display excellent contact-based antibacterial behavior against gram-negative Escherichia coli.     

Monte-carlo modeling of grain growth in Zr equal channel angular pressed and recrystallized

Grain boundary character distribution in equal-channel-angular pressed Zr was studied. Using a die design of 90°/20° and an operation temperature of 350°C. The initial grain size of 20 μm was reduced to about 270 nm with 4 passes via route B_c. The grain growth kinetics of the recrystallized state was obtained by experiment and Monte-Carlo computer simulation, respectively, which showed good agreement. Based on kinetics and morphological characteristics, it was concluded that the grain coarsening mechanism was governed by normal grain growth. No sign of abnormal grain growth was detected either in the experiment or in simulation despite taking into consideration anisotropy in grain boundary energy as well as its mobility. This indicates that grain boundaries produced by severely deformed Zr are stable against explosive coarsening. The evolution characteristics of the microstructure in the present ECA pressed and recrystallized Zr differed from those of cold rolled Ti in that the grain boundary misorientation distribution and texture were rather stable during grain growth. Jointly Appointed by the Center for Advanced Aerospace Materials This article is based on a presentation made in the 2002 Korea-US symposium on the “Phase Transformations of Nano-Materials,” organized as a special program of the 2002 Annual Meeting of the Korean Institute of Metals and Materials, held at Yonsei University, Seoul, Korea on October 25–26, 2002.