Factors Affecting Events During Oxidation of Low Density Lipoprotein: Correlation of Multiple Parameters of Oxidation

The present study shows that copper oxidation of LDL is a tightly-ordered process which can be finely controlled by appropriate selection of duration of oxidation and of concentrations of LDL and copper. Oxidation of LDL (0.1-2.0 mg LDL protein/ml) was carried out by copper catalysis (in the ratio of 2.5 μM Cu²⁺ to 0.1 mg LDL protein/ml) in phosphate-buffered saline, and was monitored by agarose gel electro-phoresis, gas chromatography (GC), anion exchange fast protein liquid chromatography (FPLC), fluorescence spectroscopy and dynamic light scattering. Analysis of the data showed strong cross correlations between many of the parameters of oxidation. Oxidation was more rapid for lower concentrations than for higher concentrations of LDL, despite the same ratio of copper to LDL being employed. Chemical kinetics analysis of the GC data suggested that 7β-hydroxycholesterol formation occurred as a first order (or pseudo first order) consecutive reaction to the oxidation of linoleate. The first order rate constants for decomposition of lioleate and production of 7β-hydroxycholesterol correlated closely with the theoretically-calculated times between collision of LDL particles. LDL particle diameter, measured by dynamic light scattering, increased by ca. 50% over 24 h oxidation, suggesting unfolding of apo B-100.

Prolonged oxidation of LDL at low concentration suggested that the radical chain reaction was able to propagate, albeit slowly, on cholesterol after all the polyunsaturated fatty acid was consumed. For higher concentrations of LDL, prolonged oxidation resulted in partial aggregation. These findings are applicable to preparing oxidised LDL with different degrees of oxidation, under controlled conditions, for studying its biological properties.     

Hexosaminidases and ganglioside catabolism in the GM2-gangliosidoses

The G_M2-gangliosidoses are a set of neurological diseases whose common features include the storage of the ganglioside G_M2, N-acetyl galactosaminyl (N-acetylneuraminyl-) galactosylglucosylceramide and related neutral glycosphingolipids in various organs (particularly brain) of affected individuals and the inability of such individuals' hexosaminidases to catalyze the hydrolysis of G_M2. Associated with this finding has been the demonstration of a deficiency in none, one or both major forms (A and B) of hexosaminidase which can be measured with artificial flurogenic or chromogenic substrates. Additionally, a deficiency in the A form of hexosaminidase which is usually associated with Tay-Sachs disease has been demonstrated in certain clinically normal adults.Recent advances in the purification of the two forms of hexosaminidase have allowed their catalytic, immunological, physical and genetic characteristics to be examined in great detail. This examination has resulted in the proposal of several models for the relationship of the hexosaminidases and their involvment in the G_M2-gangliosidoses. I discuss the evidence for these models and the implications which can be drawn from them in this review.     

Shifting white pox aetiologies affecting Acropora palmata in the Florida Keys, 1994–2014

We propose ‘the moving target hypothesis’ to describe the aetiology of a contemporary coral disease that differs from that of its historical disease state. Hitting the target with coral disease aetiology is a complex pursuit that requires understanding of host and environment, and may lack a single pathogen solution. White pox disease (WPX) affects the Caribbean coral Acropora palmata. Acroporid serratiosis is a form of WPX for which the bacterial pathogen (Serratia marcescens) has been established. We used long-term (1994–2014) photographic monitoring to evaluate historical and contemporary epizootiology and aetiology of WPX affecting A. palmata at eight reefs in the Florida Keys. Ranges of WPX prevalence over time (0–71.4%) were comparable for the duration of the 20-year study. Whole colony mortality and disease severity were high in historical (1994–2004), and low in contemporary (2008–2014), outbreaks of WPX. Acroporid serratiosis was diagnosed for some historical (1999, 2003) and contemporary (2012, 2013) outbreaks, but this form of WPX was not confirmed for all WPX cases. Our results serve as a context for considering aetiology as a moving target for WPX and other coral diseases for which pathogens are established and/or candidate pathogens are identified. Coral aetiology investigations completed to date suggest that changes in pathogen, host and/or environment alter the disease state and complicate diagnosis.     

Cluster Analysis Methods Help to Clarify the Activity—BMI Relationship of Chinese Youth

Objective: To use cluster analysis to create patterns of overall activity and inactivity in a diverse sample of Chinese youth and to evaluate their use in predicting overweight status. Research Methods and Procedures: The study populations were drawn from the 1997 and 2000 years of the longitudinal China Health and Nutrition Survey, comprised of 2702 and 2641 schoolchildren in the 1997 and 2000 cross‐sectional samples, respectively, and 1175 children in the longitudinal cohort. Cluster analysis was used to group children into nonoverlapping activity/inactivity “clusters” that were subsequently used in models of prevalent and incident overweight. Results were compared with traditional models, with activity and inactivity coded separately, to assess whether further insight was gained with the cluster analysis methodology. Results: Moderately and highly active youth were shown to have significantly decreased odds of overweight in both cross‐sectional and longitudinal analyses using cluster analysis. In incident longitudinal models, youth in the high activity/high inactivity cluster had the lowest odds of overweight [odds ratio = 0.12 (0.03, 0.44)]; in contrast, results from traditional models failed to show any significant relationship between overweight and activity or inactivity. Discussion: Cluster analysis methods allow researchers to simultaneously capture activity and inactivity in new ways. In this comparative study, only with the clustering methodology did we find a significant effect of activity on incident overweight, furthering our ability to examine this complex relationship. Interestingly, no effect of increasing levels of inactivity was observed using either method, indicating that activity seems to be the more important determinant of overweight in this population.     

Biochemical assay-based selectivity profiling of clinically relevant kinase inhibitors on mutant forms of EGF receptor

Gefitinib and erlotinib are potent EGFR tyrosine kinase inhibitors (potentially) useful for the treatment of non-small-cell lung cancer (NSCLC). Clinical responses, however, in NSCLC patients have been linked to the presence of certain activating mutations of EGFR. We used an ELISA-based biochemical assay to confirm the selective inhibitory efficacy of gefitinib and erlotinib on the activated mutant receptor. Our results are in line with the clinical observations providing evidence for the predictive power of the kinase assay. Four additional compounds were also investigated: CI-1033 and EKB-569 had dramatic inhibitory effects on all EGFR forms, whereas PD153035 and AG1478 were active on wild-type and activating mutant protein. In docking simulations with wild-type EGFR, our inhibitory data are in good agreement with the binding scores. These data confirm that anilinoquinazolines are good starting structures for the next generation of selective drugs against mutant EGFR, whereas CI-1033 and EKB-569 may represent advances for patients with both wild-type and anilinoquinazoline-resistant mutant tumors.     

Conifer colonization of a 350-year old rock fall at Lassen Volcanic National Park in northern California

Montane conifers of western North America commonly colonize mineral surfaces resulting from landslides, lahars, mudflows, and rock avalanches. This colonization can include shade-tolerant conifers that may eventually dominate the forest in a pattern termed “direct” succession. Documenting examples of this long-term successional process are useful for identifying alternative successional trajectories and indicating potential controlling mechanisms for subsequent experimental analysis. This study (1) analyzes the 1992 status of the conifer colonization on the coarse-textured surface of a 1650 AD rock avalanche in northern California and (2) measures individual growth and survivorship in permanent plots between 1992 and 2003. Increment cores of large trees indicated initial conifer colonization before 1700 AD with continuous subsequent colonization. Mean conifer density in 1992 was 725 (SD = 747) ha⁻¹ with densities increasing with decreasing rock sizes. Densities were not correlated with distances to possible seed sources. Median heights were <1 m, and the mean proportional height growth rate for healthy individuals was 0.0166 year⁻¹. The mortality rate for individuals ≥0.1-m tall was <0.007 year⁻¹. The conifers were more numerous than shrubs, and there was little apparent evidence of facilitation or inhibition of conifers. The species assemblage is mostly (89%) Abies concolor, Pinus monticola, and Pinus ponderosa individuals dispersed across an elevation range of 1870–2040 m a. s. l. This is an atypical species mix for these elevations in this location, and this mix is not readily predicted from species properties such as seed mass, seed flight distance, or drought tolerance.     

Stoichiometry of soil enzyme activity at global scale

Extracellular enzymes are the proximate agents of organic matter decomposition and measures of these activities can be used as indicators of microbial nutrient demand. We conducted a global-scale meta-analysis of the seven-most widely measured soil enzyme activities, using data from 40 ecosystems. The activities of beta-1,4-glucosidase, cellobiohydrolase, beta-1,4-N-acetylglucosaminidase and phosphatase g(-1) soil increased with organic matter concentration; leucine aminopeptidase, phenol oxidase and peroxidase activities showed no relationship. All activities were significantly related to soil pH. Specific activities, i.e. activity g(-1) soil organic matter, also varied in relation to soil pH for all enzymes. Relationships with mean annual temperature (MAT) and precipitation (MAP) were generally weak. For hydrolases, ratios of specific C, N and P acquisition activities converged on 1 : 1 : 1 but across ecosystems, the ratio of C : P acquisition was inversely related to MAP and MAT while the ratio of C : N acquisition increased with MAP. Oxidative activities were more variable than hydrolytic activities and increased with soil pH. Our analyses indicate that the enzymatic potential for hydrolyzing the labile components of soil organic matter is tied to substrate availability, soil pH and the stoichiometry of microbial nutrient demand. The enzymatic potential for oxidizing the recalcitrant fractions of soil organic material, which is a proximate control on soil organic matter accumulation, is most strongly related to soil pH. These trends provide insight into the biogeochemical processes that create global patterns in ecological stoichiometry and organic matter storage.