Trade‐off relationship between productivity and thallus toughness in Laminariales (Phaeophyceae)

Trade‐off relationships are considered key to understanding the mechanisms supporting the coexistence of multiple species within kelp beds. Thus, information on trade‐offs is expected to contribute to conservation of kelp bed diversity. To test the existence of a trade‐off between productivity and thallus toughness, thallus traits and relationships between the traits were examined for seven species of Laminariales including 24 populations. For each population, photosynthetic capacity per unit biomass (as A _mass) and nitrogen (i.e., photosynthetic nitrogen‐use efficiency, PNUE), nitrogen content (as N _mass), thallus mass per unit thallus area (as TMA) and force required to penetrate the thallus (as F _p, a common index of leaf toughness in land plants by punch test) were determined. A _mass increased with increasing N _mass. Blades with high N _mass showed high A _mass. These blades may invest a large proportion of nitrogen to the photosynthetic parts, and consequently exhibit high metabolic rates. Moreover, blades with high N _mass tended to be associated with low TMA, and N _mass decreased with increasing TMA. A significant negative correlation was observed between TMA and A _mass because of the linkage of high A _mass with high N _mass and high N _mass associated with low TMA, while a significant positive correlation was observed between TMA and F _p. The two correlations indicate the existence of a trade‐off between productivity and thallus toughness in Laminariales. PNUE showed a significant negative correlation with TMA, which also showed a significant positive correlation with F _p as the index of thallus toughness, and therefore a trade‐off relationship between productivity and thallus toughness.     

Regional Intra‐Subject Variability in Abdominal Adiposity Limits Usefulness of Computed Tomography

Objective: Computed tomography (CT) and magnetic resonance imaging, the most accurate methods of abdominal fat measurement, have been applied using a number of protocols, ranging from single‐slice area determination to multiple‐slice volume calculation. The aim of this study was to assess the validity of single‐slice CT for abdominal fat area measurement by estimating the intra‐subject variability in abdominal fat areas and comparing the ranking of subjects across four contiguous abdominal levels. Research Methods and Procedures: Nineteen premenopausal women (age, 35.3 ± 1.4 years; mean ± SE) were studied. CT was used to measure intra‐abdominal fat (IAF) area, percentage of total intra‐abdominal area (%IAF), subcutaneous abdominal fat (SAF) area, and IAF/SAF at four adjacent cross‐sectional lumbar levels (L2–L4). Intra‐subject variability (percentage) was defined as SD/mean × 100. Total body fat was measured by DXA, which was further analyzed for central abdominal fat. Results: Mean body mass index was 24.9 ± 1.0 kg/m². The average (range) intra‐subject variability was 28% (8% to 61%) for IAF, 46% (19% to 124%) for %IAF, 26% (14% to 38%) for SAF area, and 19% (7% to 71%) for IAF/SAF. The pattern of this variability was not uniform between subjects, because their ranking by IAF area was markedly different at each CT level. Discussion: We demonstrated significant intra‐subject variability in CT‐measured adipose tissue areas across four predetermined sites. This resulted in a difference in the ordering of subjects by IAF at each of the four imaging sites, suggesting that the usefulness of single‐slice CT in the assessment of abdominal adiposity in premenopausal women may be limited, particularly when performed for the purpose of making comparisons between subjects based on abdominal fat area.     

Photosynthetic gas exchange under emersed conditions in eulittoral and normally submersed members of the Fucales and the Laminariales: interpretation in relation to C isotope ratio and N and water use efficiency

Summary Ten species of brown macroalgae (five eulittoral and one submersed species of the Fucales; four submersed species of the Laminariales) from a rocky shore at Arbroath, Scotland, were examined for characteristics of emersed photosynthesis in relation to the partial pressure of CO₂ and O₂. The five eulittoral species of the Fucaceae were approaching CO₂ saturation for light-saturated photosynthesis at normal air levels of CO₂ (35 Pa) in 21 kPa O₂. The normally submersed algae are further from CO₂ saturation under these conditions, especially in the case of the four members of the Laminariales. The rate of net photosynthesis in the Fucaceae is O₂-independent in the range 2–21 kPa O₂ over the entire range of CO₂ partial pressure tested (compensation up to 95 Pa). For the other five algae tested, net photosynthesis is slightly inhibited by O₂ at 21 kPa relative to 2 kPa over the entire range of CO₂ partial pressures tested (compensation up to 95 Pa). CO₂ compensation partial pressures are low (<0.5 Pa) for the Fucaceae and independent of O₂ in the range 2–42 kPa. For the other five algae, the CO₂ compensation partial pressure are higher, and increased with O₂ partial pressure in the range 2–42 kPa. These gas exchange data show that the Fucaceae exhibit more C₄-like characteristics of their photosynthetic physiology than do the other five species tested, although even the Laminariales and Halidrys siliquosa are not classic C₃ plants in their photosynthetic physiology. These data suggest that, in emersed conditions as well as in the previously reported work on submersed photosynthesis, a CO₂ concentrating mechanism is operating which, by energized transmembrane transport of inorganic C, accumulates CO₂ at the site of RUBISCO and, at least in part, suppresses the oxygenase activity. Work with added extracellular carbonic anhydrase (CA), and with a relatively membrane-impermeant inhibitor of the native extracellular CA activity (acetazolamide), suggests that, in emersed conditions as well as in the previously reported work on algae submersed in seawater at pH 8, HCO _inf3 ^sup– is the major inorganic C species entering the cell. At optimal hydration, the rate of emersed photosynthesis in air is not less than the rate of photosynthesis when submersed in seawater, at least for the Fucaceae. ¹³C ratios of organic C for the Fucaceae are slightly more negative than is the case for the other five algae; these data are consitent with substantial (half or more of the entering inorganic C) leakage of CO₂ from the accumulated pool, and with some contribution of atmospheric CO₂ to the organic C gain by the eulittoral algae. The predicted increase in N use efficiency of photosynthesis in the Fucaceae, with their more strongly developed CO₂ concentrating mechanism, is consistent with data on emersed, but not submersed, photosynthesis for the algae collected from the wild and thus at a poorly defined N status. The more C₄-like gas exchange charateristics of photosynthesis in the eulittoral Fucaceae may be important in increasing the water use efficiency of emersed photosynthesis from the limited capital of water available for transpiration by a haptophyte.     

Alteration of fatty acid chain length of Chlamydomonas reinhardtii by simultaneous expression of medium‐chain‐specific thioesterase and acyl carrier protein

Biofuel from fatty acids with chain lengths of 8–15 (C8–C15) have properties similar to those of conventional diesel and jet fuels, thus, can save time and reduce costs for the refurbishment of engines and maintenance of oiling facilities. Most oil‐producing algae yield C16–C18 fatty acids; however, the manipulation of algae using genetic engineering is a promising approach to obtain C8–C15 fatty acids. The introduction of a medium‐chain‐specific thioesterase (TE) is expected to effectively alter algae to produce medium‐chain fatty acids (MCFAs). TE is the main determinant of fatty acid chain length as it releases fatty acids from the acyl carrier protein (ACP) in the fatty acid elongation cycle. In a previous study, the introduction of heterologous C8–C12‐specific TEs into Chlamydomonas reinhardtii did not increase the yield of MCFAs. This effect was attributed to a low affinity of the heterologous TEs to C. reinhardtii ACP. Therefore, we introduced both the C10–C14‐specific TE gene and the ACP gene from the land plant Cuphea lanceolata into C. reinhardtii. We measured free fatty acids (FFAs) and triacylglycerols (TAGs) in the transformants using liquid chromatography–mass spectrometry. The production of C12:0 and C14:0, chain length 12 and 14 without unsaturation, FFAs was not significantly increased in any of the tested strains. However, we found a slight but significant increase in TAG‐containing MCFAs in both TE only and TE–ACP transformants. The increased production rate of C14:0‐containing TAGs ranged from 1.25‐ to 1.58‐fold, indicating the ability of medium‐chain‐specific TE to increase MCFAs. These results suggest that the selection of specific TEs is important when modifying eukaryotic algae to produce MCFAs.     

Modifying fatty acid profiles through a new cytokinin‐based plastid transformation system

The widespread use of herbicides and antibiotics for selection of transgenic plants has not been very successful with regard to commercialization and public acceptance. Hence, alternative selection systems are required. In this study, we describe the use of ipt, the bacterial gene encoding the enzyme isopentenyl transferase from Agrobacterium tumefaciens, as a positive selectable marker for plastid transformation. A comparison between the traditional spectinomycin‐based aadA selection system and the ipt selection system demonstrated that selection of transplastomic plants on medium lacking cytokinin was as effective as selection on medium containing spectinomycin. Proof of principle was demonstrated by transformation of the kasIII gene encoding 3‐ketoacyl acyl carrier protein synthase III into tobacco plastids. Transplastomic tobacco plants were readily obtained using the ipt selection system, and were phenotypically normal despite over‐expression of isopentenyl transferase. Over‐expression of KASIII resulted in a significant increase in 16:0 fatty acid levels, and a significant decrease in the levels of 18:0 and 18:1 fatty acids. Our study demonstrates use of a novel positive plastid transformation system that may be used for selection of transplastomic plants without affecting the expression of transgenes within the integrated vector cassette or the resulting activity of the encoded protein. This system has the potential to be applied to monocots, which are typically not amenable to traditional antibiotic‐based selection systems, and may be used in combination with a negative selectable marker as part of a two‐step selection system to obtain homoplasmic plant lines.     

Fatty acid binding protein 7 and n‐3 poly unsaturated fatty acid supply in early rat brain development

Fatty acid binding protein 7 (FABP7), abundant in the embryonic brain, binds with the highest affinity to docosahexaenoic acid (DHA) and is expressed in the early stages of embryogenesis. Here, we have examined the consequences of the exposure to different DHA levels and of the in utero depletion of FABP7 on early rat brain development. Neurodevelopment was evaluated through the contents of two proteins, connexin 43 (Cx43) and cyclin‐dependent kinase 5 (CDK5), both involved in neuroblast proliferation, differentiation, and migration. The dams were fed with diets presenting different DHA contents, from deficiency to supplementation. DHA brain embryos contents already differed at embryonic day 11.5 and the differences kept increasing with time. Cx43 and CDK5 contents were positively associated with the brain DHA levels. When FABP7 was depleted in vivo by injections of siRNA in the telencephalon, the enhancement of the contents of both proteins was lost in supplemented animals, but FABP7 depletion did not modify phospholipid compositions regardless of the diets. Thus, FABP7 is a necessary mediator of the effect of DHA on these proteins synthesis, but its role in DHA uptake is not critical, although FABP7 is localized in phospholipid‐rich areas. Our study shows that high contents of DHA associated with FABP7 are necessary to promote early brain development, which prompted us to recommend DHA supplementation early in pregnancy. © 2015 Wiley Periodicals, Inc. Develop Neurobiol 76: 287–297, 2016     

Usefulness of Anthropometry and DXA in Predicting Intra‐abdominal Fat in Obese Men and Women

Objective: To investigate the usefulness of anthropometry and DXA in predicting intra‐abdominal fat (IAF) in obese men and women. Research Methods and Procedures: Observational, cross sectional study of 22 women and 18 men with a body mass index of 30 or above. IAF from 20 cm above and 10 cm below the L4 to L5 intervertebral disc was measured by magnetic resonance imaging (MRI) as a reference method. Central abdominal fat was measured from the upper border of L2 to the lower border of L4 by DXA. Waist and hip circumferences were also measured. Results: In obese women DXA, waist circumference and waist‐hip ratio were equally well correlated with IAF (r = 0.74, 0.75, and 0.70, respectively). In obese men DXA was moderately correlated with IAF measured by MRI (r = 0.46), whereas waist circumference and waist‐hip ratio were not significantly correlated with IAF. Discussion: The prediction of IAF in obese subjects was highly dependent on sex more than in non‐obese persons. Anthropometry and DXA were equally useful in obese women, whereas anthropometry had no predictive power and DXA was the only acceptable predictor of IAF in obese men.     

Production of 8,11‐dihydroxy and 8‐hydroxy unsaturated fatty acids from unsaturated fatty acids by recombinant Escherichia coli expressing 8,11‐linoleate diol synthase from Penicillium chrysogenum

Hydroxy unsaturated fatty acids can be used as antimicrobial surfactants. 8,11‐Linoleate diol synthase (8,11‐LDS) catalyzes the conversion of unsaturated fatty acid to 8‐hydroperoxy unsaturated fatty acid, and it is subsequently isomerized to 8,11‐dihydroxy unsaturated fatty acid by the enzyme. The optimal reaction conditions of recombinant Escherichia coli expressing Penicillium chrysogenum 8,11‐LDS for the production of 8,11‐dihydroxy‐9,12(Z,Z)‐octadecadienoic acid (8,11‐DiHODE), 8,11‐dihydroxy‐9,12,15(Z,Z,Z)‐octadecatrienoic acid (8,11‐DiHOTrE), 8‐hydroxy‐9(Z)‐hexadecenoic acid (8‐HHME), and 8‐hydroxy‐9(Z)‐octadecenoic acid (8‐HOME) were pH 7.0, 25°C, 10 g/L linoleic acid, and 20 g/L cells; pH 6.0, 25°C, 6 g/L α‐linolenic acid, and 60 g/L cells; pH 7.0, 25°C, 8 g/L palmitoleic acid, and 25 g/L cells; and pH 8.5, 30°C, 6 g/L oleic acid, and 25 g/L cells, respectively. Under these optimized conditions, the recombinant cells produced 6.0 g/L 8,11‐DiHODE for 60 min, with a conversion of 60% (w/w) and a productivity of 6.0 g/L/h; 4.3 g/L 8,11‐DiHOTrE for 60 min, with a conversion of 72% (w/w) and a productivity of 4.3 g/L/h; 4.3 g/L 8‐HHME acid for 60 min, with a conversion of 54% (w/w) and a productivity of 4.3 g/L/h; and 0.9 g/L 8‐HOME for 30 min, with a conversion of 15% (w/w) and a productivity of 1.8 g/L/h. To best of our knowledge, this is the first report on the biotechnological production of 8,11‐DiHODE, 8,11‐DiHOTrE, 8‐HHME, and 8‐HOME. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:390–396, 2017     

Population responses and fatty acid profiles of Brachionus calyciflorus (Rotifera) in relation to different thermal regimes

Diurnal temperature fluctuations affect ectothermic species more than endothermic taxa. We tested the effect of three fixed temperatures (20, 25 and 30 °C) and a 24 h variable (20–30 °C) on the population growth and fatty acid profiles of the common rotifer Brachionus calyciflorus. Depending on the temperature treatment, the peak population abundances of B. calyciflorus varied from 65 to 80 ind./ml, the lowest being on variable temperature range. The rate of population increase varied from 0.31 to 0.52 per day, highest being at 30 °C. There was a curvilinear relationship between the population density and the egg ratio (number of eggs/female) in all the tested temperature regimes. The egg ratio was higher (>0.6) for treatments involving fixed temperatures, but for variable temperature regime, the egg ratios were lower (<0.5). Temperature also induced changes in the fatty acid content of B. calyciflorus. While the total saturated fatty acids increased, both mono-unsaturated and poly-unsaturated fatty acids decreased with increasing temperature regime. These results have been interpreted in relation to the role of temperature (both intensity and mode of exposure) on the population growth characteristics of rotifers.     

Essential fatty acid profiles of maturation feeds used in freshwater ornamental fish culture

An investigation of live or freshly prepared feeds used as maturationdiets for freshwater ornamental fish was conducted to uncover similaritiesor differences in total and essential fatty acids. Analysis of these maturation feeds reveals that there are relatively low levels of totalfatty acids (0.81–8.96 mg/100 mg^–1 dry weight) and withthe exception of the beef heart diet all other feeds have undetectablelevels of docosahexaenoate (22:6n3). The beef heart diet was observed topossess 4.86 mg 100 mg^–1 dry weight of 22:6n3 most probablydue to the addition of skipjack tuna, Katsuwanus pelamis, roe. All otherfeeds examined were found to contain low to moderate levels ofeicosapentaenoate (20:5n3) 0.00–0.61 mg/100 mg^–1 dryweight. Surprisingly relatively high amounts of arachidonate (20:4n6)0.16–0.90 mg/100 mg^–1 dry weight were observed in all ofthe maturation diets and ranged between 3.79%–27.16% ona percent composition basis. The results obtained to date indicate a need toscrutinize the role of arachidonate in the maturation and spawningprocess.     

Separation of polyunsaturated fatty acids by selective inclusion in guanidinium-1,5-naphthalenedisulfonate-2-methoxyethanol host networks

Hydrogen-bonded networks composed of guanidinium (G) and 1,5-naphthalenedisulfonate (NDS) have been developed for the selective inclusion and separation of fatty acid esters based on their degree of unsaturation. Porous crystalline networks have been synthesized and include fatty acid esters during crystallization from both methanol and 2-methoxyethanol. Crystalline networks formed in methanol are selective for the inclusion of saturates in preference to polyunsaturated fatty acid methyl esters, but their applicability is limited by the competing inclusion of the solvent methanol. In 2-methoxyethanol, a three-component host structure is formed that provides 4.1 x 4.7 A2 pores which are also selective for saturated fatty acid ester inclusion. These networks do not suffer from the competing inclusion of the solvent 2-methoxyethanol as is the case with methanol, and thus complete removal of initial saturated fatty acid 2-methoxyethyl esters is possible. Binary selectivity experiments on mixtures of the 2-methoxyethyl esters of saturated stearic acid and the omega-3 fatty acid alpha-linolenic acid reveal that this three-component network structure provides nearly complete resolution of these two guests in the crystal and filtrate fractions. Separation of the 2-methoxyethyl esters of alpha-linolenic acid from the monounsaturated oleic acid is also possible, although with decreased efficiency in comparison to removal of the saturated fatty acid ester.     

Reduced blood‐brain barrier expression of fatty acid‐binding protein 5 is associated with increased vulnerability of APP/PS1 mice to cognitive deficits from low omega‐3 fatty acid diets

Lower levels of the cognitively beneficial docosahexaenoic acid (DHA) are often observed in Alzheimer's disease (AD) brains. Brain DHA levels are regulated by the blood‐brain barrier (BBB) transport of plasma‐derived DHA, a process facilitated by fatty acid‐binding protein 5 (FABP5). This study reports a 42.1 ± 12.6% decrease in the BBB transport of ¹⁴C‐DHA in 8‐month‐old AD transgenic mice (APPswe,PSEN1?E9) relative to wild‐type mice, associated with a 34.5 ± 6.7% reduction in FABP5 expression in isolated brain capillaries of AD mice. Furthermore, short‐term spatial and recognition memory deficits were observed in AD mice on a 6‐month n‐3 fatty acid‐depleted diet, but not in AD mice on control diet. This intervention led to a dramatic reduction (41.5 ± 11.9%) of brain DHA levels in AD mice. This study demonstrates FABP5 deficiency and impaired DHA transport at the BBB are associated with increased vulnerability to cognitive deficits in mice fed an n‐3 fatty acid‐depleted diet, in line with our previous studies demonstrating a crucial role of FABP5 in BBB transport of DHA and cognitive function.

     

Climate warming is predicted to reduce omega‐3, long‐chain,polyunsaturated fatty acid production in phytoplankton

Phytoplankton are the main source of energy and omega‐3 (n‐3) long‐chain essential fatty acids (EFA) in aquatic ecosystems. Their growth and biochemical composition are affected by surrounding environmental conditions, including temperature, which continues to increase as a result of climate warming. Increasing water temperatures may negatively impact the production of EFA by phytoplankton through the process of homeoviscous adaptation. To investigate this, we conducted an exploratory data synthesis with 952 fatty acid (FA) profiles from six major groups of marine and freshwater phytoplankton. Temperature was strongly correlated with a decrease in the proportion of n‐3 long‐chain polyunsaturated FA (LC‐PUFA) and an increase in omega‐6 FA and saturated FA. Based on linear regression models, we predict that global n‐3 LC‐PUFA production will be reduced by 8.2% for eicosapentaenoic acid (EPA) and 27.8% for docosahexaenoic acid (DHA) with an increase in water temperature of 2.5 °C. Using a previously published estimate of the global production of EPA by diatoms, which contribute to most of the world's supply of EPA, we predict a loss of 14.2 Mt of EPA annually as a result of ocean warming. The n‐3 LC‐PUFA are vitally important for an array of key physiological functions in aquatic and terrestrial organisms, and these FA are mainly produced by phytoplankton. Therefore, reduced production of these EFA, as a consequence of climate warming, is predicted to negatively affect species that depend on these compounds for optimum physiological function. Such profound changes in the biochemical composition of phytoplankton cell membranes can lead to cascading effects throughout the world's ecosystems.     

Quantitative trait loci mapping for conjugated linoleic acid,vaccenic acid and ∆9‐desaturase in Italian Brown Swiss dairy cattle using selective DNA pooling

A selective DNA pooling approach was applied to identify QTL for conjugated linoleic acid (CLA), vaccenic acid (VA) and Δ⁹‐desaturase (D9D) milk content in Italian Brown Swiss dairy cattle. Milk samples from 60 animals with higher values (after correction for environmental factors) and 60 animals with lower values for each of these traits from each of five half‐sib families were pooled separately. The pools were genotyped using the Illumina BovineSNP50 BeadChip. Sire allele frequencies were compared between high and low tails at the sire and marker level for SNPs for which the sires were heterozygous. An r procedure was implemented to perform data analysis in a selective DNA pooling design. A correction for multiple tests was applied using the proportion of false positives among all test results. BTA 19 showed the largest number of markers in association with CLA. Associations between SNPs and the VA and Δ⁹‐desaturase traits were found on several chromosomes. A bioinformatics survey identified genes with an important role in pathways for milk fat and fatty acids metabolism within 1 Mb of SNP markers associated with fatty acids contents.     

Modulation of adipocyte differentiation by omega‐3 polyunsaturated fatty acids involves the ubiquitin‐proteasome system

We have evaluated the effects of three different omega‐3 polyunsaturated fatty acids (ω‐3 PUFAs) – docosahexaenoic acid (DHA), eicosapentaenoic acid (EPA) and docosapentaenoic acid (DPA) on fat accumulation and expression of adipogenic and inflammatory markers using both 3T3‐L1 pre‐adipocytes and differentiated 3T3‐L1 adipocytes. Our results indicate that ω‐3 PUFAs induce the degradation of fatty acid synthase through the ubiquitin‐proteasome system, which is likely to have beneficial metabolic effect on adipose cells. Omega‐3 PUFAs also increase overall levels of polyubiquitinated proteins, at least in part through decreasing the expression of proteasome subunits. Moreover, adipocytes are resistant to proteasome inhibition, which induces adipophilin while decreasing perilipin expression. On the other hand, ω‐3 PUFAs decrease expression of SREBP1 while inducing expression of adipophilin and GLUT4. Moreover, all three ω‐3 PUFAs appear to induce tumour necrosis factor‐α without affecting NFκB levels. All three ω‐3 PUFAs appear to have overall similar effects. Further research is needed to elucidate their mechanism of action.