Finite Element Analysis of Simulations of Cancellous Bone Resorption

A stochastic simulation of the resorption of cancellous bone has been developed and integrated with a finite element model to predict the resultant change in structural properties of bone as bone density decreases. The resorption represents the net imbalance of osteoclast and osteoblast activity that occurs in osteoporosis. A simple lattice structure of trabecular bone is considered, with an examination of the lattice geometry and discretization indicating that just five trabeculae need to be modelled. The results from the analysis show how the mechanical properties of the cancellous bone degrade with osteoporosis and demonstrate how the method can be used to predict the relationships between stiffness and density or porosity.     

Two levels of resting potential in cardiac purkinje fibers

In an appropriate ionic environment, the resting potential of canine cardiac purkinje fibers may have either of two value. By changing the external K concentration, [K](0), in small steps, it was shown that, in the low (1 mM) Cl, Na-containing solutions used in this study, the two levels of resting potential could be obtained only within a narrow range of [K](0) values; that range was usually found between 1 and 4 mM. Within the critical [K](0) range the resting potential could be shifted from either level to the other by the application of small current pulses. It was shown that under these conditions the steady-state current- voltage relationship was “N-shaped,” and that a region of both negative slope, and negative chord conductance lay between the two stable zero-current potentials. The negative chord conductance was largely due to inward sodium current, only part of which was sensitive to tetrodotoxin (TTX). Under appropriate conditions, the negative chord conductance could be abolished by several experimental interventions and the membrane potential thereby shifted from the lower to the higher resting level: those interventions which were effective by presumably diminishing the steady-state inward current included reducing the external sodium concentration, adding TTX, or adding lidocaine; those which presumably increased the steady-state outward current included small increases in [K](0), brief depolarizations to around -20 mV, or the addition of acetylcholine chloride.     

The importance of weather and agronomic factors for the overwinter survival of yellow rust (Puccinia striiformis) and subsequent disease risk in commercial wheat crops in England

Disease survey data from 4475 randomly selected crops of wheat from England and Wales during 1985–2000 showed that yellow rust was most prevalent in 1988, 1989, 1990, 1998 and 1999. Disease severity on the upper two leaves was low as >95% crops had received foliar fungicides. Factors affecting the presence or absence (incidence) of yellow rust were investigated using random effects logistic regression (general linear mixed model). This enabled crop management (risk) variables for individual crops to be combined with meteorological variables measured at the county level. Two models are presented that analysed the effect of host genotype on incidence either solely through yellow rust resistance rating (Model 1) or by including both resistance rating (fixed effect) and cultivar (fitted as a random term) (Model 2). In both models, the percentage of crops with yellow rust decreased with cultivar disease resistance ratings ≥3, the occurrence of severe frosts (P < 0.05) of timing of fungicide sprays, previous cropping or summer weather. The use of risk variables associated with overwintering survival may help adjust fungicide inputs to seasonal risk.     

Low reduction potential of Ero1alpha regulatory disulphides ensures tight control of substrate oxidation

Formation of disulphide bonds within the mammalian endoplasmic reticulum (ER) requires the combined activities of Ero1α and protein disulphide isomerase (PDI). As Ero1α produces hydrogen peroxide during oxidation, regulation of its activity is critical in preventing ER-generated oxidative stress. Here, we have expressed and purified recombinant human Ero1α and shown that it has activity towards thioredoxin and PDI. The activity towards PDI required the inclusion of glutathione to ensure sustained oxidation. By carrying out site-directed mutagenesis of cysteine residues, we show that Ero1α is regulated by non-catalytic disulphides. The midpoint reduction potential (E°′) of the regulatory disulphides was calculated to be approximately −275 mV making them stable in the redox conditions prevalent in the ER. The stable regulatory disulphides were only partially reduced by PDI (E°′~−180 mV), suggesting either that this is a mechanism for preventing excessive Ero1α activity and oxidation of PDI or that additional factors are required for Ero1α activation within the mammalian ER.     

Effects of Ca- and Na-lignosulfonate on starch gelatinization and network formation

The interaction of lignosulfonates with starches was examined by microscopy and viscosity measurements. 8% starch dispersions with Ca- or Na-lignosulfonate, or with only Ca²⁺ or Na⁺, were heated to 97 °C and cooled to 50 °C in a Brabender Viscograph, the gelatinization was followed by light microscopy and image analysis, and the gel network formed after cooling to 4 °C was studied under the transmission electron microscope.

The lignosulfonates (2%) delayed the initial granule swelling in all starches (native maize, waxy maize and waxy barley). The presence of ions enhanced amylose leakage resulting in lower peak viscosity. The viscosity during cooling increased more with Ca-LS than with Na-LS. With a low lignosulfonate concentration the network formed after cooling was homogeneous with fine strands. With Na-lignosulfonate, as well as with Na⁺, the network connectivity deteriorated and spherical aggregates formed. Ca-lignosulfonate induced a network with thick strands, but with Ca²⁺ the strands became thinner.     

A DNA vaccine against tuberculosis based on the 65 kDa heat-shock protein differentially activates human macrophages and dendritic cells

Background

Application of plasmid DNA for immunization of food-producing animals established new standards of food safety. The addition of foreign products e.g. pDNA into the food chain should be carefully examined to ensure that neither livestock animals nor consumers develop unpredicted or undesirable side-effects.

Methods

A quantitative real-time PCR (QRTPCR) methodology was developed to study the biodistribution and persistence of plasmid DNA vaccine pDNAX (pVAX-Hsp60 TM814) in mice and beef cattle. The linear quantification range and the sensitivity of the method was found to be 10 – 10⁹ copies per reaction (500 ng/gDNA) and 3 copies per reaction, respectively.

Results

Persistence of pDNAX in mice muscle tissue was restricted to injection site and the amount of pDNAX showed delivery formulation dependent (naked pDNA, electroporation, cationic liposome complexes) and mouse age-dependent clearance form injection site but pDNAX was still detectable even after 365 days. The QRTPCR analysis of various muscle tissue samples of vaccinated beef bulls performed 242–292 days after the last revaccination proved that residual pDNAX was found only in the injection site. The highest plasmid levels (up to 290 copies per reaction) were detected in the pDNAX:CDAN/DOPE group similarly to mice model. No pDNA was detected in the samples from distant muscles and draining lymph nodes.

Conclusion

Quantitative real-time PCR (QRTPCR) assay was developed to assess the residual pDNA vaccine pVAX-Hsp60 TM814 in mice and beef cattle. In beef cattle, ultra low residual level of pDNA vaccine was only found at the injection site. According to rough estimation, consumption of muscles from the injection site represents almost an undetectable intake of pDNA (400 fg/g muscle tissue) for consumers. Residual plasmid in native state will hardly be found at measurable level following further meat processing. This study brings supportive data for animal and food safety and hence for further approval of pDNA vaccine field trials.     

A simple assay for a human serum phospholipase A2 that is associated with high-density lipoproteins

Phospholipase A2 (PLA2) activity is usually assayed with expensive radioactive or chromogenic substrates unsuitable for performing large numbers of assays. We have designed a simple microplate assay for human serum PLA2 using the chromogenic substrate 4-nitro-3-octanoyloxy-benzoic acid. Using this substrate, serum PLA2 activity was similar to that measured with the previously characterized chromogenic phospholipid substrate 1,2-bis-heptanoylthio-glycerophosphocholine. However, the assay described here appears to be more sensitive. The mean PLA2 activity in serum from healthy volunteers (n = 30) measured by this assay was 10.4 +/- 1.6 micromol x h(-1) x ml(-1). The assay is reproducible and is suitable for the analysis of large numbers of samples in a clinical setting. We have also demonstrated that 94% of the PLA2 activity in normal human serum is associated with high-density lipoproteins and that serum PLA2 activity is positively correlated with the lipoprotein parameters total triglyceride (P < 0.0001), total cholesterol (P < 0.0001), and atherogenic index (P = 0.008). The serum PLA2 activity was calcium dependent and was inhibited by the serine protease inhibitor 3,4-dichloroisocoumarin (EC(50) = 0.4 mM). The PLA2 activity characterized here is unlikely to be due to plasma platelet-activating factor acetylhydrolase or low molecular weight His-Asp sPLA2, and may represent a new sPLA2 type.     

Utilisation of morphological features in life table studies of Liriomyza huidobrensis (Dipt., Agromyzidae) developing in lettuce

Abstract: A method of distinguishing different larval instars of Liriomyza huidobrensis morphologically, using measurements of the cephalopharyngeal skeleton was developed. The growth ratios of cephalopharyngeal skeletons between first and second and second and third instar larvae were 1.80 and 1.47, respectively, enabling clear separation to be achieved for experimental work. Using this method the development rates of the immature stages of L. huidobrensis feeding on Lactuca sativa were determined under constant temperatures of 11, 16, 19, 26 and 28 ± 1°C and were shown to increase linearly with temperature over the range investigated. The theoretical lower threshold temperatures for development from oviposition to the end of each larval instar or pupal stage were 5.35, 6.30, 6.20 and 5.70°C, respectively. The overall threshold temperature for development from oviposition to 50% adult emergence (5.70°C) was used to calculate degree‐day (DD) requirements for development from oviposition to each larval instar or pupal eclosion, which were 84.3, 30.1, 58.9, 143.7 DD, respectively. The use of these data for optimizing the timing of application of control agents which are effective against specific developmental stages is discussed.