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.     

Effect of progressive systemic sclerosis on antral myoelectrical activity and gastric emptying

In patients with progressive systemic sclerosis (PSS) suffering from chronic dyspepsia the stomach may be affected by this disease. The objective of this study was to investigate both antral myoelectrical activity and gastric emptying in PSS patients. Electrogastrography (EGG) was performed in 17 PSS patients (16 female, one male, median age 58 years, range 32-74 years) with chronic dyspepsia. After an overnight fast during one hour in the fasting and one hour in the fed state after ingestion of a liquid-solid test meal (370 kcal; liquid phase labeled with 0.5 mCi 99mTc-colloid) antral electrical activity was measured by one pair of electrodes sonographically placed on the skin overlying the gastric antrum. Several EGG parameters including dominant frequency (DF), percentages of DF in the normal range (2-4 cycles per minute [cpm]), bradygastria (< 2 cpm) and tachygastria (4-10 cpm), dominant frequency instability coefficient (DFIC), and postprandial to preprandial power ratio (PR) were calculated. The data were correlated to results obtained in 20 age- and gender-matched healthy subjects. In addition, the data were compared to gastric retention of the radionuclide at 60 min measured by simultaneous scintigraphy. The PSS patients did not reveal electrical disturbances. They even exhibited a significant postprandial decrease in DFIC, bradygastria, and tachygastria (ns) compared to healthy subjects. Over 50% of the PSS patients showed a delayed gastric emptying. However, EGG did not correlate to radioscintigraphy significantly. Our results reflect an absent relationship between antral myoelectrical activity in EGG and gastric emptying. Therefore, electrogastrography is unsuitable to assess gastric involvement in PSS.     

Effect of hemofiltration on hemodynamics and systemic concentrations of anaphylatoxins and cytokines in human sepsis

OBJECTIVE: To determine whether hemofiltration (HF) can eliminate cytokines and complement components and alter systemic hemodynamics in patients with severe sepsis. DESIGN: Prospective observation study. SETTING: Surgical intensive care unit of a university hospital. PATIENTS: 16 patients with severe sepsis. INTERVENTIONS: Continuous zero-balanced HF without dialysis (ultrafiltrate rate 2 l/h) was performed in addition to pulmonary artery catheterization, arterial cannulation, and standard intensive care treatment. MEASUREMENTS AND MAIN RESULTS: Plasma and ultrafiltrate concentrations of cytokines (the interleukins IL-1 beta, IL-6, IL-8, and tumor necrosis factor alpha) and of complement components (C3adesArg, C5adesArg) were measured after starting HF (t0) and 4 h (t4) and 12 h later (t12). Hemodynamic variables including mean arterial pressure (MAP), mean central venous pressure, mean pulmonary artery pressure, pulmonary capillary wedge pressure, and cardiac output were serially determined. During HF, cytokine plasma concentrations remained constant. However, C3adesArg and C5adesArg plasma concentrations showed a significant decline during 12-h HF (C3adesArg: t0 = 676.9 +/- 99.7 ng/ml vs t12 = 467.8 +/- 71, p < 0.01; C5adesArg: 26.6 +/- 4.7 ng/ml vs 17.6 +/- 6.2, p < 0.01). HF resulted in a significant increase over time in systemic vascular resistance (SVR) and MAP (SVR at t0: 669 +/- 85 dyne.s/cm5 vs SVR at t12: 864 +/- 75, p < 0.01; MAP at t0: 69.9 +/- 3.5 mmHg vs MAP at t12: 82.2 +/- 3.7, p < 0.01). CONCLUSIONS: HF effectively eliminated the anaphylatoxins C3adesArg and C5adesArg during sepsis. There was also a significant rise in SVR and MAP during high volume HF. Therefore, HF may represent a new modality for removal of anaphylatoxins and may, thereby, deserve clinical testing in patients with severe sepsis.     

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.     

Polarisation transformer on Ti:LiNbO3 with reset-free optical operation for heterodyne/homodyne receivers

Experimental results for an integrated-optical state-of-polarisation (SOP) controller device on Ti:LiNbO3 for continuous broadband optical operation, with small temperature sensitivity and the possibility of integration with polarisation- insensitive directional couplers, are presented.     

Structural and functional meta-analytic evidence for fronto-subcortical system deficit in progressive supranuclear palsy

Meta-analytic methods were used to determine the most sensitive indexes to fronto-subcortical deficit in progressive supranuclear palsy (PSP) and to further characterize the neurocognitive and related features of PSP that can provide a basis of comparison to other disorders with prominent subcortical brain lesions. Studies dating back to 1984 were gathered and calibrated to compare the neuropsychological, neuroimaging, and neurophysiological test results from 229 patients with PSP, and 357 healthy controls. The tests most sensitive to fronto-subcortical deficit in PSP were mostly neuropsychological measures that include such tests as the Stroop Task, Trail Making Test Part A, and Purdue pegboard performance. We conclude that although neuropsychological measures may be most sensitive to deficits in PSP, they are also less specific and valid indicators of fronto-subcortical brain system integrity.     

Congenital chylothorax in siblings

We describe two cases of congenital chylothorax in siblings with important differences from previously described familial cases. Our findings support the likelihood of an autosomal recessive inheritance in some cases of this condition, rather than X-linked recessive inheritance, which has also been suggested. Autopsy findings from one of these cases and others previously described suggest that the pathophysiological mechanisms involved may be variable.