An in vitro biomechanical comparison between prototype tapered shaft cortical bone screws and AO cortical bone screws for an equine metacarpal dynamic compression plate fixation of osteotomized equine third metacarpal bones

OBJECTIVES: To compare biomechanical properties of a prototype 5.5 mm tapered shaft cortical screw (TSS) and 5.5 mm AO cortical screw for an equine third metacarpal dynamic compression plate (EM-DCP) fixation to repair osteotomized equine third metacarpal (MC3) bones. STUDY DESIGN: Paired in vitro biomechanical testing of cadaveric equine MC3 with a mid-diaphyseal osteotomy, stabilized by 1 of 2 methods for fracture fixation. ANIMAL POPULATION: Adult equine cadaveric MC3 bones (n=12 pairs). METHODS: Twelve pairs of equine MC3 were divided into 3 groups (4 pairs each) for (1) 4-point bending single cycle to failure testing, (2) 4-point bending cyclic fatigue testing, and (3) torsional single cycle to failure testing. An EM-DCP (10-hole, 4.5 mm) was applied to the dorsal surface of each, mid-diaphyseal osteotomized, MC3 pair. For each MC3 bone pair, 1 was randomly chosen to have the EM-DCP secured with four 5.5 mm TSS (2 screws proximal and distal to the osteotomy; TSS construct), two 5.5 mm AO cortical screws (most proximal and distal holes in the plate) and four 4.5 mm AO cortical screws in the remaining holes. The control construct (AO construct) had four 5.5 mm AO cortical screws to secure the EM-DCP in the 2 holes proximal and distal to the osteotomy in the contralateral bone from each pair. The remaining holes of the EM-DCP were filled with two 5.5 mm AO cortical screws (most proximal and distal holes in the plate) and four 4.5 mm AO cortical screws. All plates and screws were applied using standard AO/ASIF techniques. Mean test variable values for each method were compared using a paired t-test within each group. Significance was set at P<.05. RESULTS: Mean 4-point bending yield load, yield bending moment, bending composite rigidity, failure load and failure bending moment of the TSS construct were significantly greater (P<.00004 for yield and P<.00001 for failure loads) than those of the AO construct. Mean cycles to failure in 4-point bending of the TSS construct was significantly greater (P<.0002) than that of the AO construct. The mean yield load and composite rigidity in torsion of the TSS construct were significantly greater (P<.0039 and P<.00003, respectively) than that of the AO construct. CONCLUSION: The TSS construct provides increased stability in both static overload testing and cyclic fatigue testing. CLINICAL RELEVANCE: The results of this in vitro study support the conclusion that the EM-DCP fixation using the prototype 5.5 mm TSS is biomechanically superior to the EM-DCP fixation using 5.5 mm AO cortical screws for the stabilization of osteotomized equine MC3.     

Mycotoxins and other secondary metabolites produced in vitro by Penicillium paneum Frisvad and Penicillium roqueforti Thom isolated from baled grass silage in Ireland

Secondary metabolites produced by Penicillium paneum and Penicillium roqueforti from baled grass silage were analyzed. A total of 157 isolates were investigated, comprising 78 P. paneum and 79 P. roqueforti isolates randomly selected from more than 900 colonies cultured from bales. The findings mostly agreed with the literature, although some metabolites were not consistently produced by either fungus. Roquefortine C, marcfortine A, and andrastin A were consistently produced, whereas PR toxin and patulin were not. Five silage samples were screened for fungal metabolites, with two visually moldy samples containing up to 20 mg/kg of roquefortine C, mycophenolic acid, and andrastin A along with minor quantities (0.1-5 mg/kg) of roquefortines A, B, and D, festuclavine, marcfortine A, and agroclavine. Three visually nonmoldy samples contained low amounts of mycophenolic acid and andrastin A. The ability of both molds to produce a diverse range of secondary metabolites in vitro and in silage should be a concern to livestock producers.     

Potency- and Selectivity-Enhancing Mutations of Conotoxins for Nicotinic Acetylcholine Receptors Can Be Predicted Using Accurate Free-Energy Calculations

Nicotinic acetylcholine receptor (nAChR) subtypes are key drug targets, but it is challenging to pharmacologically differentiate between them because of their highly similar sequence identities. Furthermore, α-conotoxins (α-CTXs) are naturally selective and competitive antagonists for nAChRs and hold great potential for treating nAChR disorders. Identifying selectivity-enhancing mutations is the chief aim of most α-CTX mutagenesis studies, although doing so with traditional docking methods is difficult due to the lack of α-CTX/nAChR crystal structures. Here, we use homology modeling to predict the structures of α-CTXs bound to two nearly identical nAChR subtypes, α3β2 and α3β4, and use free-energy perturbation (FEP) to re-predict the relative potency and selectivity of α-CTX mutants at these subtypes. First, we use three available crystal structures of the nAChR homologue, acetylcholine-binding protein (AChBP), and re-predict the relative affinities of twenty point mutations made to the α-CTXs LvIA, LsIA, and GIC, with an overall root mean square error (RMSE) of 1.08 ± 0.15 kcal/mol and an R² of 0.62, equivalent to experimental uncertainty. We then use AChBP as a template for α3β2 and α3β4 nAChR homology models bound to the α-CTX LvIA and re-predict the potencies of eleven point mutations at both subtypes, with an overall RMSE of 0.85 ± 0.08 kcal/mol and an R² of 0.49. This is significantly better than the widely used molecular mechanics—generalized born/surface area (MM-GB/SA) method, which gives an RMSE of 1.96 ± 0.24 kcal/mol and an R² of 0.06 on the same test set. Next, we demonstrate that FEP accurately classifies α3β2 nAChR selective LvIA mutants while MM-GB/SA does not. Finally, we use FEP to perform an exhaustive amino acid mutational scan of LvIA and predict fifty-two mutations of LvIA to have greater than 100X selectivity for the α3β2 nAChR. Our results demonstrate the FEP is well-suited to accurately predict potency- and selectivity-enhancing mutations of α-CTXs for nAChRs and to identify alternative strategies for developing selective α-CTXs.     

Effects of non-cropped landscape diversity on spatial dynamics of farmland birds in intensive farming systems

Increasing habitats diversity in agricultural landscapes has been proposed as a key measure for reversing the decline of farmland biodiversity in Europe. However, indicators used for assessing such a potential compensation effect usually only rely on species diversity and abundance while ignoring variations in species-specific vulnerability. The extent to which habitat diversity may reverse the decline of specialist species in Europe to farming systems is thus still unclear. In this study, we investigate whether the effect of non-cropped habitat diversity on farmland birds’ occurrences was dependent on species’ specialization for habitats. In particular, we focused on the relative effects of non-cropped habitat diversity on species’ abilities to persist or to colonize new vacant areas. We used a capture-recapture statistical framework to study the spatial dynamics of 20 farmland bird species in France monitored from 2001 to 2007. We found that non-cropped landscape diversity reduces both the probabilities that a species becomes extinct locally and that a species colonizes new vacant areas, and the occupancy rate. Although this suggests a possible stabilizing effect of the surrounding habitat diversity on species occurrence in farming systems, the occupancy was only weakly affected. Moreover, we found that the most specialist species were the more negatively affected by this landscape diversity in terms of colonization abilities. We argue that accounting for the differences in habitat specialization among farmland species can improve conservation policies dedicated to the management of landscape diversity.     

Effect of the ionic strength of pulsed electric field treatment medium on the physicochemical and structural characteristics of lactoferrin

Pulsed electric field (PEF) treatment (35 kV cm(-1) for 19.2 μs using bipolar 2 μs pulses) was conducted on bovine lactoferrin (LF; 0.4 mg mL(-1)) prepared in simulated milk ultrafiltrate (SMUF), at concentrations between 0.2× and 2× normal strength, with electrical conductivities ranging from 0.17 to 1.04 S m(-1). The physicochemical and structural characteristics (LF content by a spectrophotometric and an ELISA method, surface hydrophobicity, electrophoretic mobility, far-UV circular dichroism spectra, and tryptophan fluorescence) of LF dissolved in SMUF of all strengths tested were not changed after PEF treatment. The PEF treatment of LF in 0.2 strength SMUF did not cause the release of LF-bound ferric ion into the aqueous phase, with a concentration of LF-bound iron being the same as that of the untreated LF control (174 μg L(-1)). However, in treatment media with higher ionic strengths, ferric ion was released from the LF molecule into the aqueous phase. The concentration of LF-bound iron decreased from 174 μg L(-1) for the LF treated in 0.2 strength SMUF to 80 μg L(-1) for that treated in double-strength SMUF. The results suggest that the PEF-induced iron depletion of LF does not appear to cause an appreciable conformational change in LF molecules. PEF treatment could be developed as a novel physical way to produce iron-depleted LF, as an alternative to the existing chemical method.     

Evolution of chemical and sensory properties during aging of top-fermented beer

The aging and consequent changes in flavor molecules of a top-fermented beer were studied. Different aging conditions were imposed on freshly bottled beer. After 6 months of aging, the concentration changes were recorded for acetate esters, ethyl esters, carbonyls, Maillard compounds, dioxolanes, and furanic ethers. For some flavor compounds, the changes with time of storage were monitored at different temperatures, either with CO(2) or with air in the headspace of the bottles. For some molecules a relationship was determined between concentration changes and sensory evaluation results. A decrease in volatile esters was responsible for a reduced fruity flavor during aging. On the contrary, various carbonyl compounds, some ethyl esters, Maillard compounds, dioxolanes, and furanic ethers showed a marked increase, due to oxidative and nonoxidative reactions. A very high increase was found for furfural, 2-furanmethanol, and especially the furanic ether, 2-furfuryl ethyl ether (FEE). For FEE a flavor threshold in beer of 6 mug/L was determined. In the aged top-fermented beer, FEE concentrations multiple times the flavor threshold were observed. This was associated with the appearance of a typical solvent-like flavor. As the FEE concentration increased with time at an almost constant rate, with or without air in the headspace, FEE (and probably other furanic ethers) is proposed as a good candidate to evaluate the thermal stress imposed on beer.     

Techniques for pollution monitoring in remote sites: II. Ozone

An O₃ monitoring system for use in remote sites lacking AC line power was developed. A chemiluminescent O₃ monitor, ethylene delivery system, and a DC power supply were combined to allow unattended O₃ measurements over extended periods of time. A Campbell Scientific 21X micrologger in combination with a relay was used to control the ethylene delivery solenoid and O₃ analyzer power, and to record results. The system was tested over a six week period in late summer 1990 and gave results comparable to those from surrounding sites.     

Die „Baumfuß-Methode”︁, dargestellt anhand einer Untersuchung der Immissionsbelastung von Nordwest-Jugoslawischen Buchenwäldern

The “Trunk Base Method”, Demonstrated by an Investigation on Immission Load of Beech Stands in Northwestern Yugoslavia Due to stemflow, the soil area close to the stem of old beech trees (Fagus sylvatica) differs from the soil farther from the stem as a consequence of higher input rates of all elements. It is postulated that the level of deposition determines the degree of deviation in soil chemical properties of the trunk base area (“Baumfußbereich” = “micro-habitat”) compared with soil which is not influenced by stemflow water (“Vergleichsstandort” = “macro-habitat”). The paper attempts to estimate the relative load of air pollution in 10 beech stands (mainly Rendzinas and Cambisols from limestone and dolomite) of northwestern Yugoslavia. In each plot 10 micro- and macro-habitats were chosen, for analysis of soil samples of Ah horizons for pH and exchangeable cations after percolation with NH₄Cl. The differences in pH, base saturation and saturation of (H+Fe) between trunk base area and related soil are used to determine acid loading. The saturation of (H+Fe) turned out to be the most appropriate indicator to describing the acidification. The results indicate high acid deposition rates at sites 1–5,9 and 10, but low rates at stands 6–8. It is assumed, that the main sources of pollutants reaching the plots 1–5, which are located at the western declivity of Risnjak, are industrial plants in northern Italy and towns at the Adriatic coast. The sources of pollutants reaching the sites 9 and 10, which are located at the eastern declivity of Gorski Kotar, respectively Lika region, are the industrial plants in northern Yugoslavia and Hungary.     

Chemical composition and antimicrobial activity of the essential oils from the gum of Turkish pistachio (Pistacia vera L.)

The essential oil from the gum of Pistachio (Pistacia vera L. (Anacardiaceae)) grown in Turkey was obtained by the hydro-distillation method, and its chemical composition was analyzed by GC and GC-MS. Moreover, the antimicrobial activities of the oil against the growth of 13 bacteria and 3 pathogenic yeasts were evaluated using the agar-disk diffusion and minimum inhibitory concentration (MIC) methods. The results showed that the essential oil contained about 89.67% monoterpenes, 8.1% oxygenated monoterpenes and 1.2% diterpenes. alpha-Pinene (75.6%), beta-pinene (9.5%), trans-verbenol (3.0%), camphene (1.4%), trans-pinocarveol (about 1.20%), and limonene (1.0%) were the major components. The antimicrobial results showed that the oil inhibited nine bacteria and all the yeasts studied, and the activities were considerably dependent upon concentration and its bioactive compounds such as carvacrol, camphene, and limonene. Moreover, the essential oil of the gum was found to be more effective yeastcide than Nystatin, synthetic yeastcide. Furthermore, the antibacterial activities of the oil were lower than those of standard antibiotics, ampicillin sodium, and streptomycine sulfate under the conditions studied.     

Interaction of maize roots and rhizosphere microorganisms

The interaction between the roots of Zea mays L. and its rhizosphere microflora was studied in nutrient solution with respect to two exemplary aspects - water soluble vitamins and the four most frequent carbohydrates. The exudation of vitamins and sugars into the medium is increased in a solid substrate (glass-ballotini simulating soil structure) in comparison with nutrient solution only. The amounts of vitamins released (1 to 421 ng per plant) are high enough for a pronounced influence on the rhizosphere microflora to be expected. However, the microbial population feeding on the exudates has very simple nutritional demands. In spite of appreciable amounts of vitamins found in the inoculated nutrient solution, the growth of vitamin requiring species is not stimulated compared to the soil population. The roots are mostly inhabited by shorter rods and occasionally by actinomycete-like filaments. The percentage of the root area covered by its microflora, assuming a monolayer, is 4% in the root region where root hairs are just emerging, 7% in the root hair zone and 20% in the oldest part of the roots, 10 mm below the grain. In the presence of rhizosphere microorganisms, root dry weight is lower than that of axenically grown plants because the diameter of the primary root is diminished. In addition, the vitamin and sugar content of the root is affected by rhizosphere microorganisms. The results can be interpreted as an indication that phytohormones may participate in interactions between roots and bacteria.