A novel family VII esterase with industrial potential from compost metagenomic library

<p>Abstract</p> <p>Background</p> <p>Among the vast microbial genomic resources now available, most microbes are unculturable in the laboratory. A culture-independent metagenomic approach is a novel technique that circumvents this culture limitation. For the screening of novel lipolytic enzymes, a metagenomic library was constructed from compost, and the clone of <it>estCS2 </it>was selected for lipolytic properties on a tributyrin-containing medium.</p> <p>Results</p> <p>The <it>estCS2 </it>sequence encodes a protein of 570 amino acid residues, with a predicted molecular mass of 63 kDa, and based on amino acid identity it most closely matches (45%) the carboxylesterase from <it>Haliangium ochraceum </it>DSM 14365. EstCS2 belong to family VII, according to the lipolytic enzyme classification proposed by Arpigny and Jaeger, and it retains the catalytic triad Ser₂₄₅-Glu₃₆₃-His₄₆₆that is typical of an α/β hydrolase. The Ser₂₄₅residue in the catalytic triad of EstCS2 is located in the consensus active site motif GXSXG. The EstCS2 exhibits strong activity toward <it>p</it>-nitrophenyl caproate (C6), and it is stable up to 60°C with an optimal enzymatic activity at 55°C. The maximal activity is observed at pH 9, and it remains active between pH 6-10. EstCS2 shows remarkable stability in up to 50% (v/v) dimethyl sulfoxide (DMSO) or dimethylformamide (DMF). The enzyme has the ability to cleave sterically hindered esters of tertiary alcohol, as well as to degrade polyurethanes, which are widely used in various industries.</p> <p>Conclusions</p> <p>The high stability of EstCS2 in organic solvents and its activity towards esters of ketoprofen and tertiary alcohols, and in polyurethane suggests that it has potential uses for many applications in biotransformation and bioremediation.</p

Transmit Power Allocation for a Modified V-BLAST System

This paper was presented in part at the 13th IEEE Symposium on Personal, Indoor and Mobile Radio Communications, Lisbon, Portugal, September 2002.Abstract—In this letter, we present a modification of Vertical Bell Laboratories Layered Space–Time (V-BLAST), and propose an effective transmit power allocation (TPA) scheme for the modified system. The proposed TPA scheme minimizes the uncoded biterror rate (BER) averaged over all detection stages, and requires small feedback overhead. Simulation results show that the modified V-BLAST system with the proposed TPA scheme provides a significant reduction in the uncoded BER compared with the conventional V-BLAST system. When the minimum mean-square error nulling is adopted, the modified V-BLAST system is found to achieve the uncoded BER performance comparable to that of the maximum-likelihood detection for the conventional V-BLAST architecture.This work was supported in part by the Brain Korea 21 Project

Different Susceptibilities to Low Temperature Photoinhibition in the Photosynthetic Apparatus Among three Cultivars of Cucumber (Cucumis sativus L.)

Susceptibility to low temperature photoinhibition in photosynthetic apparatus was compared among three cucumber cultivars, Gahachungjang (GH), Banbaekjijeo (BB) and Gaeryangsymji (GR). By chilling in the light for 6 h, a sustained decrease in the potential quantum yield (Fv/Fm) and the oxidizable P700 contents was observed, and the decrease was less in GH than in BB and GR. Although the difference was small, some Φ PSII remained in GH after light-chilling for 6 h indicating that a few electrons can flow around photosystem II (PSII). As a consequence, the primary electron acceptor of PSII, Q A , was reduced slowly and was not fully reduced after lightchilling for 6 h in GH. Although the amplitude was small, the development of NPQ was also faster in GH, indicating a higher capacity for non-photochemical energy dissipation. The relative fraction of a fast relaxing component of NPQ (qf) was higher in GH. After light-chilling for 5 h, the values of qf in BB and GR became much smaller than that in GH, indicating BB and GR suffered more significant uncoupling of ATPase and/or irreversible damages in PSII. When fluorescence induction transients were recorded after chilling, significant differences in quenching coefficients (qQ and qN) were observed among the three cultivars

THE EFFECTS OF WHEELCHAIR CAMBER AND HANDRIM SIZE IN WHEELCHAIR BASKETBALL MOVEMENT

Improving the wheelchair design can be an important determinant of high performance in the wheelchair basketball. Researchers have focused on increasing the efficiency of the wheelchair, looking at the factors such as the seat position, handrim size, and wheel camber. Most handrim and wheel chamber studies, however, have focused only on the propulsive phase. Quick turn as well as fast propulsion is essential to the success in a wheelchair basketball game and the energy efficiency becomes particularly important in a prolonged wheelchair use. The purpose of this study was to investigate the effects of wheelchair camber and handrim size on the linear propulsion, turn velocity, and efficiency

5′-Triphosphate-RNA-independent activation of RIG-I via RNA aptamer with enhanced antiviral activity

RIG-I is a cytosolic receptor for non-self RNA that mediates immune responses against viral infections through IFNα/β production. In an attempt to identify novel tools that modulate IFNα/β production, we used SELEX technology to screen RNA aptamers that specifically target RIG-I protein. Most of the selected RIG-I aptamers contained polyU motifs in the second half regions that played critical roles in the activation of RIG-I-mediated IFNβ production. Unlike other known ligands, RIG-I aptamer bound and activated RIG-I in a 5′-triphosphate-independent manner. The helicase and RD domain of RIG-I were used for aptamer binding, but intact RIG-I protein was required to exert aptamer-mediated signaling activation. Furthermore, replication of NDV, VSV and influenza virus in infected host cells was efficiently blocked by pre- or post-treatment with RIG-I aptamer. Based on these data, we propose that RIG-I aptamer has strong potential to be an antiviral agent that specifically boosts the RIG-I-dependent signaling cascade

Antiplatelet and Antithrombotic Activity of a Traditional Medicine, Hwangryunhaedok-Tang

Platelet activation and accumulation at the site of vascular injury are central to thrombus formation resulted in thrombotic disorders. Medicinal herbs could be one of the most important pharmaceutical agents that ameliorate thrombotic disorders, such as unstable angina, myocardial infarction, stroke, and peripheral vascular diseases. Hwangryunhaedok-tang (HRT) is a traditional herbal medicine that displays multiple biological properties including anti-inflammatory abilities. However, its role in platelet activation has not been fully studied. Hence, we examined whether HRT has a potent inhibitory effect on platelet aggregation and thrombus formation. We demonstrated that HRT (30, 50, and 100 μg/ml) significantly impaired thrombin- and collagen-related peptide-induced platelet aggregation, granule secretion, thromboxane B2 generation, and intracellular Ca2+ mobilization. Biochemical studies revealed that HRT is involved in inhibiting the phosphorylation of phospholipase C and protein kinase B. The oral administration of HRT (30, 50, and 100 mg/kg once daily for 1 and/or 7 days) efficiently ameliorates ferric chloride induced arterial thrombus formation in vivo. Tail bleeding time was not significantly increased. The qualitative phytochemical constituents of the HRT extract were investigated using high-performance liquid chromatography. Our results demonstrated that HRT shows potential antiplatelet and antithrombotic effects without affecting hemostasis. Hence, HRT could be an effective therapeutic agent for the treatment of thrombotic diseases

Stress-induced Cardiomyopathy during Pulmonary Resection (Takotsubo Syndrome) - A case report -

Stress-induced cardiomyopathy is caused by emotional or physical stressors and mimics acute myocardial infarction, though Stress-induced cardiomyopathy is characterized by reversible left ventricular (LV) apical ballooning in the absence of significant coronary artery disease. We describe a 51-year-old male who underwent left upper lobectomy for non-small cell lung cancer, and during which cardiogenic arrest occurred due to stress-induced cardiomyopathy, successfully managed by intra-aortic balloon pumping and extracorporeal membrane oxygenation

An Isolated True Aneurysm of the Superficial Femoral Artery in a Young Woman - A case report -

A 39-year-old woman was admitted to the hospital due to a pulsatile mass on her right inner thigh that was evident for two months. She did not exhibit any risk factors of atherosclerosis, no evidence of vasculitis, or any signs of previous trauma history. Ultrasound and computed tomography revealed an adult fist-sized aneurysm on the distal superficial femoral artery. The aneurysm was resected and peripheral circulation was restored with the interposition of a saphenous vein graft. The resected aneurysm had three layers that showed atherosclerosis on histological examination

Structural and Biochemical Bases for the Inhibition of Autophagy and Apoptosis by Viral BCL-2 of Murine γ-Herpesvirus 68

All gammaherpesviruses express homologues of antiapoptotic B-cell lymphoma-2 (BCL-2) to counter the clearance of infected cells by host antiviral defense machineries. To gain insights into the action mechanisms of these viral BCL-2 proteins, we carried out structural and biochemical analyses on the interactions of M11, a viral BCL-2 of murine γ-herpesvirus 68, with a fragment of proautophagic Beclin1 and BCL-2 homology 3 (BH3) domain-containing peptides derived from an array of proapoptotic BCL-2 family proteins. Mainly through hydrophobic interactions, M11 bound the BH3-like domain of Beclin1 with a dissociation constant of 40 nanomole, a markedly tighter affinity compared to the 1.7 micromolar binding affinity between cellular BCL-2 and Beclin1. Consistently, M11 inhibited autophagy more efficiently than BCL-2 in NIH3T3 cells. M11 also interacted tightly with a BH3 domain peptide of BAK and those of the upstream BH3-only proteins BIM, BID, BMF, PUMA, and Noxa, but weakly with that of BAX. These results collectively suggest that M11 potently inhibits Beclin1 in addition to broadly neutralizing the proapoptotic BCL-2 family in a similar but distinctive way from cellular BCL-2, and that the Beclin1-mediated autophagy may be a main target of the virus