Design and Characterization of a New pVII Combinatorial Phage Display Peptide Library for Protease Substrate Mining Using Factor VII Activating Protease (FSAP) as Model

We describe a novel, easy and efficient combinatorial phage display peptide substrate-mining method to map the substrate specificity of proteases. The peptide library is displayed on the pVII capsid of the M13 bacteriophage, which renders pIII necessary for infectivity and efficient retrieval, in an unmodified state. As capture module, the 3XFLAG was chosen due to its very high binding efficiency to anti-FLAG mAbs and its independency of any post-translational modification. This library was tested with Factor-VII activating protease (WT-FSAP) and its single-nucleotide polymorphism variant Marburg-I (MI)-FSAP. The WT-FSAP results confirmed the previously reported Arg/Lys centered FSAP cleavage site consensus as dominant, as well as reinforcing MI-FSAP as a loss-of-function mutant. Surprisingly, rare substrate clones devoid of basic amino acids were also identified. Indeed one of these peptides was cleaved as free peptide, thus suggesting a broader range of WT-FSAP substrates than previously anticipated.     

Occupational Factors and 5-Year Weight Change Among Men in a Danish National Cohort.

This study explored whether factors related to the work environment could predict changes in body mass index (BMI) and whether the effect of psychosocial factors was dependent on baseline BMI. The sample consisted of 1,980 male employees from the Danish National Work Environment Cohort Study. Changes in BMI between 1995 and 2000 were analyzed, by multiple regression, as a function of background variables and a series of occupational variables obtained in 1995. Age, baseline BMI, job insecurity, and psychological demands predicted changes in BMI. Job insecurity and high or low psychological demands increased the likelihood of weight gain among obese employees, whereas they increased the likelihood of weight loss among employees with a low BMI. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

The ultrasound appearances of neonatal renal vein thrombosis

Renal vein thrombosis (RVT) is the most frequently occurring vascular condition in the new-born kidney. The predisposing factors include dehydration, sepsis, birth asphyxia, maternal diabetes, polycythaemia and the presence of an indwelling umbilical venous catheter. (RVT) may present clinically with a flank mass, haematuria, hypertension or renal failure. Many imaging modalities have been employed, but ultrasound is the technique most commonly used in the evaluation of neonates with suspected RVT. Thrombosis commences in the small renal veins and subsequently propagates via larger interlobar veins to the main renal vein and inferior vena cava (IVC). The ultrasound appearances depend upon the stage at which the examination is performed and extent of the thrombus. Initially, the interlobular and interlobar thrombus appears as highly echogenic streaks. These streaks commence in a peripheral, focal segment of the involved kidney and only persist for a few days. In the first week the affected kidney swells and becomes echogenic with prominent echopoor medullary pyramids. Later, the swelling increases and the kidney becomes heterogenous with loss of corticomedullary differentiation. Grey scale ultrasound readily demonstrates thrombus within the renal vein and IVC. Adrenal haemorrhage is a recognized association and may be identified ultrasonically. Colour Doppler scanning provides additional information. In the early stages of RVT, colour Doppler may demonstrate absent intrarenal and renal venous flow. Ultimately, the kidney may recover, show focal scarring or become atrophic. Thus, ultrasound provides an accessible and reliable tool in the assessment of suspected neonatal RVT.     

Scarring trachoma is associated with polymorphism in the tumor necrosis factor alpha (TNF-alpha) gene promoter and with elevated TNF-alpha levels in tear fluid

Tumor necrosis factor alpha (TNF-alpha) may play a central role in the disease pathogenesis which occurs as a consequence of chlamydial infection. To investigate the importance of TNF-alpha gene promoter polymorphisms and TNF-alpha levels in tear fluid in scarring trachoma, a large matched-pair case-control study was performed in The Gambia. The -308A allele was present in a higher proportion of patients (28.4%) than controls (18.4%), with an increasing association for homozygotes (chi2 for trend, P = 0.032; allele frequency, 0.163 in patients and 0.099 in controls; chi2, P = 0.025). For the -238A allele, the association was similar but not significant. The disease association was highly significant when the number of either -308A or -238A sites in an individual was considered (P = 0.003). TNF-alpha promoter alleles are tightly linked to some HLA class I and II alleles, but multivariate analysis confirmed that the disease associations were independent of HLA, although a class I allele, A*6802, is also associated with disease. TNF-alpha was more frequently detected in tear samples from patients (27.6%) than from controls (15.9%), increasingly so for higher levels of detectable TNF-alpha (P = 0.015). Among patients, detectable TNF-alpha in tears was highly associated with the presence of ocular chlamydial infection (P < 0.001). The results indicate that TNF-alpha plays a major role in the tissue damage and scarring which occurs as a consequence of Chlamydia trachomatis infection.     

Removal of nitrates from groundwater through the use of artificial wetlands

Intensive fertilizer use has led to nitrate contamination of groundwater in many areas. This article examines the possibility of removing nitrates through the construction of artificial wetlands. A pilot study conducted in Denmark has found that the method is effective and economical, at least for small waterworks, though further treatment is necessary before an acceptable microbiological water quality is attained.     

Semiparametric Estimation in Time‐Series Regression with Long‐Range Dependence

Abstract. We consider semiparametric estimation in time‐series regression in the presence of long‐range dependence in both the errors and the stochastic regressors. A central limit theorem is established for a class of semiparametric frequency domain‐weighted least squares estimates, which includes both narrow‐band ordinary least squares and narrow‐band generalized least squares as special cases. The estimates are semiparametric in the sense that focus is on the neighbourhood of the origin, and only periodogram ordinates in a degenerating band around the origin are used. This setting differs from earlier studies on time‐series regression with long‐range dependence, where a fully parametric approach has been employed. The generalized least squares estimate is infeasible when the degree of long‐range dependence is unknown and must be estimated in an initial step. In that case, we show that a feasible estimate which has the same asymptotic properties as the infeasible estimate, exists. By Monte Carlo simulation, we evaluate the finite‐sample performance of the generalized least squares estimate and the feasible estimate.     

Sludge reed bed facilities: operation and problems.

Short operating periods and problems with dewatering efficiency, vegetation, mineralisation and odour are primarily caused by incorrect construction of the filter, poor capillary connections, an inadequate number of basins, insufficient basin area and overloading during commissioning and during subsequent operation. Dimensioning should be based on the sludge quality including the dewatering qualities and not solely on calculations of the sludge production. Loading after planting depends on the development level of the vegetation. The loading program should ensure that reed establishment is not impeded and should prevent the sludge residue from growing so quickly horizontally and vertically that the reeds cannot colonise the sludge residue. Overloading results in an anaerobic sludge residue with ensuing methane production. Typically, a sludge reed bed facility with a loading period of maximum 5 days, must be built with 10 basins to permit a rest phase of about 40 days. Facilities with 8 basins, where it is possible to load 1 basin for 7 days, will be able to have a 7-week rest phase before the first basin is loaded again. Facilities with for example 8-13 basins and loading period of between 4 and 10 days are able to achieve rest periods of up to 2 to 2(1/2) months, which results in the optimal evapotranspiration and minerilization. The conditions and thus the possibilities vary depending upon the type of sludge.