Micellization of bovine beta-casein studied by isothermal titration microcalorimetry and cryogenic transmission electron microscopy

The association behavior, critical micellization concentration (CMC), and enthalpy of demicellization (DeltaHdemic) of bovine beta-casein were studied, for the first time by isothermal titration calorimetry, in a pH 7.0 phosphate buffer with 0.1 ionic strength and in pure water. In the buffer solutions, the CMC decreased asymptotically from 0.15 to 0.006 mM as the temperature was raised from 16 to 45 degrees C. DeltaHdemic decreased with increasing temperature between 16 and 28 degrees C but increased from 28 to 45 degrees C. Thermodynamic analysis below 30 degrees C is consistent with the Kegeles shell model, which suggests a stepwise association process. At higher temperatures, this model exhibits limitations, and the micellization becomes much more cooperative. The CMC values in water, measured between 17 and 28 degrees C, decreased with increasing temperature and, expectedly, were higher than those found in the buffer solutions. beta-Casein micelles were visualized and characterized, for the first time in their hydrated state, using advanced digital-imaging cryogenic transmission electron microscopy. The images revealed small, oblate micelles, about approximately 13 nm in diameter. The micelles shape and dimensions remained nearly constant in the temperature range of 24-35 degrees C.     

The effect of land use type on net nitrogen mineralization on abandoned agricultural land: Silver birch stand versus grassland

The effect of land use type on the dynamics and annual rate of net nitrogen mineralization (NNM) in a naturally generated silver birch stand and in a grassland, both on abandoned agricultural land, was assessed in situ in the upper 0–20 cm soil layer using the method of buried polyethylene bags. Annual NNM rate in the birch stand (156 kg N ha⁻¹ year⁻¹) was higher than in the grassland (102 kg N ha⁻¹ year⁻¹); in both cases NNM covered a major part of the plants annual nitrogen demand. The rate of NNM in the upper 0–10 cm soil layer in the birch stand (99 kg N ha⁻¹ year⁻¹) exceeded the respective rate of NNM in the grassland (51 kg N ha⁻¹ year⁻¹) roughly two times. In the grassland the rates of NNM in the 0–10 and 10–20 cm layers were equal; in the birch stand NNM in the 0–10 cm layer was 1.7 times higher than in deeper 10–20 cm layer. The intensity of daily NNM in the upper 0–10 cm soil layer in the birch stand was the highest in June and in the grassland in May, 776 and 528 mg kg⁻¹ N day⁻¹, respectively. In our study no significant correlation was found between NNM and the environmental factors monthly mean soil temperature, soil moisture content and pH.     

Early growth and development of silver birch (<Emphasis Type="Italic">Betula pendula</Emphasis> Roth.) plantations on abandoned agricultural land

Eight-year-old plantations (11) of silver birch were studied on abandoned agricultural land in Estonia. Trees were planted on uncovered soil and on polyethylene. After eight growing seasons, the height and diameter at breast height of the trees grown on polyethylene were significantly (P < 0.001) larger than corresponding parameters of the trees grown on uncovered soil. The annual height increment of the birches grown on the polyethylene was significantly (P < 0.05) larger during six growing seasons after planting compared to that of the trees grown on uncovered soil. However, annual height increments were not significantly different in the 7th and the 8th growing seasons. The use of polyethylene mulching had a statistically significant effect (P < 0.001) on the height of the beginning of the live crown in 8-year-old plantations. The differences between the values of live crown ratio of the trees grown on polyethylene and the values of live crown ratio of the trees grown on uncovered soil decreased during six growing seasons. However, by the 7th and 8th growing seasons, there was no significant difference (P > 0.05) between the values of the live crown ratio. The height growth of silver birch grown without mulching as well as with mulching was found to be more intensive on Glossic Podzoluvisol, Calcaric Cambisol, Calcaric Luvisol and Dystric Gleysol; however, the height growth was more intensive on mulched soil. The height growth of the birches was modelled on the single-tree and stand levels for five soil types.     

Self-assembly of bovine beta-casein below the isoelectric pH

Beta-casein is an intrinsically unstructured amphiphilic protein that self-assembles into micelles at neutral pH. This paper reports that beta-casein self-organizes into micelles also under acidic conditions. The protein association behavior and micelle characteristics at pH 2.6, well below the p I, are presented. The pH was found to strongly affect the micelle shape and dimensions. Cryogenic transmission electron microscopy (cryo-TEM) experiments revealed disk-like micelles of 20-25 nm in length and approximately 3.5 nm in height in acidic conditions. An aggregation number of 6 was determined by sedimentation equilibrium under these conditions. Isothermal titration calorimetry experiments verified the association below the p I and allowed determination of the micellization enthalpy, the critical micellar concentration, and the micellization relative cooperativity (MR). Small-angle X-ray scattering results at concentrations below the critical micellization concentration (CMC) suggest that the monomeric protein is likely in a premolten globule state at low pH. Calculations of the protein charge at acidic and neutral pH reveal a similar high net charge but considerable differences in the charge distribution along the protein backbone. Overall the results show that beta-casein is amphiphilic at low pH, but the distribution of charge along the protein chain creates packing constraints that affect the micelle organization, leading at concentrations above the CMC to the formation of disk micelles.     

A field investigation of the use of the pedometer for the early detection of lameness in cattle

The efficacy of the pedometer to predict lameness earlier than the appearance of the clinical signs in a herd of dairy cows was investigated by correlating pedometric activity (PA) with clinical cases of lameness. The computer program was set to identify cows with a reduction of 5% or more in PA compared with their own previous 10 days average; these animals were then examined for clinical lameness. At the same time, every lame cow was checked to see if and when its PA was reduced. Forty-six cows showed a reduced PA; 38 cases of lameness were identified by either a reduction in PA or clinical observation; of these, 21 lame cows (45.7%) showed a reduction in PA of 5% or more, 7 to 10 days prior to the appearance of clinical signs. This cohort comprised 55.3% of the lame cows. In 92% of the lame cows identified by PA, the decrease was above 15%.     

Molecular diagnosis of mango malformation disease and phylogeny of Fusarium mangiferae

Mango malformation disease (MMD) caused by various Fusaria, including the fungus Fusarium mangiferae, is difficult to diagnose and cannot be controlled effectively. In this study a PCR diagnostic tool was developed for detection of F. mangiferae by generating primers which flank fungus-specific sequences. The assay is suitable for both in-vitro and in-vivo tests, and is capable of detecting 10?pg of the fungal DNA. Relatively high genetic variation was discovered between various isolates of the pathogen based on amplified fragment length polymorphism (AFLP) analyses. In addition, some isolates of F. mangiferae (including the accession numbers FMS-123 and NRRL 25226), were found to be quite distinct from most F. mangiferae accessions. Likewise, representative MMD-associated Fusaria from the Asian and American clades of the Gibberella fujikuroi species complex were further differentiated by AFLP analyses.     

Genetic association between DNA fingerprint fragments and loci controlling agriculturally important traits in avocado (Persea americana Mill.)

Summary Associations between DNA fingerprint fragments and 16 avocado quantitative trait loci were tested for in two avocado tamilies by one way analysis of variance and multiple regression analysis. Based on these two analyses the fragments P4, P8, E2 and E5 in Pinkerton × Ettinger progeny were found to be associated with harvest duration, skin color, skin thickness and skin surface, respectively. The fragments P1, P8, B1 and B4 in the Pinkerton × Bacon progeny (half sibs of the original population) were found to be associated with fruit weight, skin color, seed size and peeling, respectively. Based on the original Pinkerton progeny as well as on the half sibs progeny, the fragment P8 was found to be associated with black-purple fruit skin color.The fragment pattern intensity of DNA pools of progeny having either green or black-purple fruit skin color —supported these data. These results are interpreted as genetic-linkage between the DNA fingerprint fragment P8 and locas (1) regulating fruit skin color in avocado.     

Water quality,trend detection,and causality

This paper begins by discussing the detection of a trend in water quality time series. Using the method of Box and Jenkins, an example of trend identification in sediment loading in the Iowa River is presented. Subsequent to the identification of a trend, an approach is suggested whereby the factors giving rise to the trend can be detected. For the example being considered, it is indicated that the cropping practices in the watershed have a significant impact on the detected trend. Thus, each 1% increase in the area of corn and soybeans planted results in about a 0.42% increase in sediment loading.