An endo-acting proline-specific oligopeptidase from Treponema denticola ATCC 35405: evidence of hydrolysis of human bioactive peptides

An endo-acting proline-specific oligopeptidase (prolyl oligopeptidase [POPase], EC 3.4.21.26) was purified to homogeneity from the Triton X-100 extracts of cells of Treponema denticola ATCC 35405 (a human oral spirochete) by a procedure that comprised five successive fast protein liquid chromatography steps. The POPase is a cell-associated 75- to 77-kDa protein with an isoelectric point of ca. 6.5. The enzyme hydrolyzed (optimum pH 6.5) the Pro-pNA bond in carbobenzoxy-Gly-Pro-p-nitroanilide (Z-Gly-Pro-pNA) and bonds at the carboxyl side of proline in several human bioactive peptides, such as bradykinin, substance P, neurotensin, angiotensins, oxytocin, vasopressin, and human endothelin fragment 22-38. The minimum hydrolyzable peptide size was tetrapeptide P3P2P1P'1, while the maximum substrate size was ca. 3 kDa. An imino acid residue in position P1 was absolutely necessary. The hydrolysis of Z-Gly-Pro-pNA was potently inhibited by the following, with the Ki(app) (in micromolar) in parentheses: insulin B-chain (0.7), human endothelin-1 (0.5), neuropeptide Y (1.7), substance P (32.0), T-kinin (4.0), neurotensin (5.0), and bradykinin (16.0). Chemical modification and inhibition studies suggest that the POPase is a serine endopeptidase whose activity depends on the catalytic triad of COOH ... Ser ... His but not on a metal. The amino acid sequence around the putative active-site serine is Gly-Gly-Ser-Asn-Pro-Gly. The enzyme is suggested to contain a reactive cysteinyl residue near the active site. Amino acid residues 4 to 24 of the first 24 N-terminal residues showed a homology of 71% with the POPase precursor from Flavobacterium meningosepticum and considerable homology with the Aeromonas hydrophila POPase. The ready hydrolysis of human bioactive peptides at bonds involving an imino acid residue suggests that enzymes like POPase may contribute to the chronicity of periodontal infections by participating in the peptidolytic processing of those peptides.     

Molecular characterization of erythrocyte glucose-6-phosphate dehydrogenase deficiency in Al-Ain District, United Arab Emirates

Operational skills involved in controlling a motor vehicle were measured in two groups of very healthy elderly drivers and a young control group to test the hypothesis that there are age-related declines in operational performance that may influence driver safety. An actual behind-the-wheel, standardized road test was employed using a motor vehicle equipped with sensors to record speed, braking activity, and lane position, as well as direction and magnitude of front-wheel and eye-movement excursions. The data from these sensors were used as dependent measures of operational performance. Older drivers made fewer steering and eye-movement excursions and drifted across the center line more frequently than the young control group. Younger drivers drove significantly faster and executed more braking applications than did their older counterparts. The motor-vehicle operational performance of older healthy drivers was related to visual-spatial attentional declines and the useful field of vision associated with the normal aging process.     

Effects of changes in strain path on the anisotropy of yield stresses of low-carbon steel and 70-30 brass sheets

Uniaxial tensile tests in various directions following uniaxial extension, equibiaxial stretching or plane strain rolling have been performed to study the effects of changes in strain path on the anisotropy of yield stresses of aluminium-killed low-carbon steel and 70-30 brass sheets. The anisotropy could be predicted from the specimen textures, if dislocation structure were equiaxed, as in the case of equibiaxial stretching. However, elongated dislocation cell structures, developed in the steel specimens prestrained in uniaxial tension or plane strain rolling, gave rise to the second-stage yield stresses higher than predicted from textures in the directions different from the maximum prestrain direction. Planar dislocation structures in the brass specimens prestrained in uniaxial tension or plane strain rolling gave the second-stage yield stresses lower than predicted from the textures in the directions different from the maximum prestrain direction. The phenomena are discussed based on textures and dislocation structures.     

Pharmacological specificities and modalities of prescriptions for corticoids for asthmatic adults

The efficacy of corticosteroids in asthma has been recognized over 40 years ago. Since that time, the advent of inhaled forms has further improved the therapeutic of these drugs which are now recognized as the fundamental treatment for asthma, and described in detail by national and international consensus. Based on a large body of literature, it can now be recommended to prescribe inhaled corticosteroids for symptomatic asthma patients. Long-term treatment is required and dosage not exceeding 1000 micrograms/d (beclometasone dipropionate equivalent) in adults are safe. Differences in the pharmacological characteristics of the various systematic and inhaled corticosteroids can be used to adapt treatment and administration route to each patient and achieve good patient compliance with optimal therapeutic efficacy.     

An isolated ZVS/ZCS flyback converter using the leakage inductanceof the coupled inductor

This paper describes a simple and effective way to modify an existing hard-switched flyback power converter into a circuit with zero-voltage switching (ZVS) and zero-current switching (ZCS). The key improvement is to turn the unattractive features of the coupled inductor leakage inductance and snubber capacitor into attractive ones. The coupled inductor leakage inductance and snubber are used to form a quasi-resonant circuit to facilitate ZVS/ZCS of all power devices. The operating principles of the power converter and experimental results are presented     

Anomalous variations of OFF-State leakage current in poly-Si TFTunder static stress

Studies the anomalous variations of the OFF-state leakage current (I_OFF) in n-channel poly-Si thin-film transistors (TFTs) under static stress. The dominant mechanisms for the anomalous I_OFF can be attributed to (1) I_OFF increases due to channel hot electrons trapping at the gate oxide/channel interface and silicon grain boundaries and (2) I_OFF decreases due to hot holes accumulated/trapped near the channel/bottom oxide interface near the source region. Under the stress of high drain bias, serious impact ionization effect will occur to generate hot electrons and hot holes near the drain region. Some of holes will be injected into the gate oxide due to the vertical field (~(V_Gstress V_Dstress)/T _OX) near the drain and the others will be migrated from drain to source along the channel due to lateral electric field (~V_Dstress/L_CH)     

Effects of quinine on the mechanical frequency response of the cupula in the fish lateral line

Quinine induces changes in the motion of the cupula in the lateral line canal of the African knife-fish in response to sinusoidal water movements. Two different phases in the action of quinine on the cupular frequency response can be discerned. In the first phase the best frequency, i.e., the frequency at which the cupular vibratory displacement is maximal in response to constant-amplitude sinusoidal canal fluid displacement, shifts toward higher frequencies. During this phase, lasting about 70-100 min, the best frequency increases by a factor between 1.3 and 1.5. In the second phase, during roughly the following 90 min, the best frequency decreases gradually to a value 0.3-0.5 times that observed before the application of quinine.     

A region-based selective optical flow back-projection for genuine motion vector estimation

Motion vector plays one significant feature in moving object segmentation. However, the motion vector in this application is required to represent the actual motion displacement, rather than regions of visually significant similarity. In this paper, region-based selective optical flow back-projection (RSOFB) which back-projects optical flows in a region to restore the region's motion vector from gradient-based optical flows, is proposed to obtain genuine motion displacement. The back-projection is performed based on minimizing the projection mean square errors of the motion vector on gradient directions. As optical flows of various magnitudes and directions provide various degrees of reliability in the genuine motion restoration, the optical flows to be used in the RSOFB are optimally selected based on their sensitivity to noises and their tendency in causing motion estimation errors. In this paper a deterministic solution is also derived for performing the minimization and obtaining the genuine motion magnitude and motion direction.