Fatty Acid Production and Direct Acyl Transfer through Polar Lipids Control TAG Biosynthesis during Nitrogen Deprivation in the Halotolerant Alga Dunaliella tertiolecta

The aims of this work were to evaluate the contribution of the free fatty acid (FA) pool to triacylglyceride (TAG) biosynthesis and to try to characterize the mechanism by which FA are assimilated into TAG in the green alga Dunaliella tertiolecta. A time-resolved lipidomic analysis showed that nitrogen (N) deprivation induces a redistribution of total lipidome, particularly of free FA and major polar lipid (PL), in parallel to enhanced accumulation of polyunsaturated TAG. The steady-state concentration of the FA pool, measured by prolonged ¹⁴C-bicarbonate pre-labeling, showed that N deprivation induced a 50% decrease in total FA level within the first 24 h and up to 85% after 96 h. The abundance of oleic acid increased from 50 to 70% of total free FA while polyunsaturated FA (PUFA) disappeared under N deprivation. The FA flux, measured by the rate of incorporation of ¹⁴C-palmitic acid (PlA), suggests partial suppression of phosphatidylcholine (PC) acyl editing and an enhanced turnover of the FA pool and of total digalactosyl-diacylglycerol (DGDG) during N deprivation. Taken together, these results imply that FA biosynthesis is a major rate-controlling stage in TAG biosynthesis in D. tertiolecta and that acyl transfer through PL such as PC and DGDG is the major FA assimilation pathway into TAG in that alga and possibly in other green microalgae. Increasing the availability of FA could lead to enhanced TAG biosynthesis and to improved production of high-value products from microalgae.     

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.     

Expression of relaxin receptor LRG7, canine relaxin, and relaxin-like factor in the pelvic diaphragm musculature of dogs with and without perineal hernia

OBJECTIVES: To compare the expression of canine relaxin, relaxin-like factor (RLF), and relaxin receptors within the muscles of the pelvic diaphragm of dogs with perineal hernia (PH) and clinically normal dogs. STUDY DESIGN: In vivo comparative study. ANIMALS: Fifteen client-owned intact male dogs with PH were studied. Four mature intact male dogs with no evidence of perineal pathology served as controls. METHODS: Biopsy samples from the levator ani, coccygeus, and internal obturator muscles were obtained. RNA samples were reverse transcribed and analyzed by real-time PCR for the expression of canine relaxin receptor LRG7, relaxin, and RLF. RESULTS: Significantly higher expression levels of canine relaxin receptors occurred in the musculature of the pelvic diaphragm and internal obturator muscle in dogs with PH compared with normal dogs. Expression of canine RLF revealed no significant difference between dogs with PH and controls. The difference in the expression of canine relaxin between groups was not statistically significant. CONCLUSIONS: Relaxin receptor up-regulation occurs in the coccygeus, levator ani, and internal obturator muscles of dogs with PH. CLINICAL RELEVANCE: The higher expression of relaxin receptors within the muscles of the pelvic diaphragm in dogs with PH suggests that relaxin might play a role in the pathogenesis of PH. Atrophy of these muscles, which predisposes to PH, may be attributable to increased relaxin activity.     

Effect of irrigation water salinity on transpiration and on leaching requirements: A case study for bell peppers

Maximization of crop yields when the salinity of irrigation water is high depends on providing plant transpiration needs and evaporative losses, as well as on maintaining minimum soil solution salinity through leaching. The effect of the amount of applied irrigation water was studied regarding transpiration, yields, and leaching fractions as a function of irrigation water salinity. Bell pepper (Capsicum annum L. vars. Celica and 7187) in protected growing environments in the Arava Valley of Israel was used as a case study crop to analyze water quantity–salinity interactions in a series of lysimeter, field and model simulation experiments. Leaching fraction was found to be highly influenced by plant feedback, as transpiration depended on root zone salinity. Increased application of saline irrigation water led to increased transpiration and yields. The higher the salinity level, the greater the relative benefit from increased leaching. The extent of leaching needed to maximize yields when irrigating with saline water may make such practice highly unsustainable.     

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%.