Unsaturated Fatty Acid Synthesis During the Development of Isolated Sunflower (Helianthus annuus L.) Seeds

The effect of temperature on unsaturated fatty acid synthesisin developing sunflower seed embryos (Helianthus annuus L.)has been studied using isolated seeds grown in culture. Variabilitybetween individual embryos in the response to temperature wasalso investigated. Oil and dry matter accumulation in cultured embryos were similarto those of embryos allowed to develop in intact plants, andthe effect of increasing temperature in lowering the amountof linoleic acid in seed oil was reproduced in cultured embryos.The isolated seed culture system, therefore, constitutes a suitablemodel system for studies of oil synthesis in developing sunflowerembryos. The decrease in linoleic acid synthesis in response to highertemperature was detectable after only 18 day-degrees incubation,and the incorporation of labelled substrates suggests that alterationsin the fatty acid composition of seed oil in response to temperatureare produced by an effect on the desaturation of newly synthesizedoleate rather than through turnover of existing lipid. Variation in fatty acid composition between individual embryosgrown at constant temperature was considerable. The detectionof embryos with high linoleic acid levels following growth athigh temperature indicates that potential may exist for theselection of cultivars for temperature-stable fatty acid compositionin sunflower oil. Key words: Fatty acid synthesis, Helianthus annuus, Sunflower seeds     

Distribution of Fibronectin and Collagen during Mouse Limb and Palate Development

Indirect immunofluorescence has been used to study the distribution of fibronectin and collagen types I, II, and III in the developing primary and secondary palatal processes and forelimb buds of the Swiss Webster (NIH) mouse. In the palatal processes fibronectin and types I and III collagen are distributed throughout the mesenchyme. Fibronectin is present in the basement membrane, while types I and III collagen are localized in a linear, discontinuous fashion beneath the basement membrane. Fibronectin is not observed in the epithelium, including the presumptive fusion areas. In the forelimb bud these components show a similar distribution prior to chondrogenesis (early day 11). When chondrogenesis commences (late day 11 or early day 12) fibronectin and, to a lesser degree, types I and III collagen are apparently concentrated in the core mesenchyme, suggesting that fibronectin has a role in initiating chondrogenesis, perhaps by increasing cellular aggregation. Type II collagen is observed only in chondrogenic regions. The codistribution of fibronectin and types I and III collagen supports in vitro studies which indicate that cells use fibronectin to bind to collagen in the matrix. The developing chondrogenic regions appear to lose fibronectin gradually, concomitant with the appearance of type II collagen, suggesting that fibronectin is not involved in the maintenance of functional chondrocytes in their matrices.     

Differential lipid reserves influence host-seeking behaviour in the mosquitoes Aedes cantans and Aedes punctor

ipid reserves of bait-caught female Ae.cantans and Ae.punctor mosquitoes (Diptera: Culicidae) were significantly higher than in teneral females. Female Ae.cantans given access to 10% w/v sucrose solution post-emergence showed an ability to synthesize lipid and, after 192 h, they were willing to take a bloodmeal from a human volunteer. At this point, mean lipid reserves were not significantly different from mean lipid reserves of bait-caught females. Prior to 192 h, females would not take a bloodmeal and lipid reserves were significantly lower than in bait-caught females. Female Ae.punctor given access to 10% w/v sucrose solution post-emergence also showed an ability to synthesize lipid. Females of this species were willing to feed from a human host after only 48 h, at which point lipid content was not significantly different from that in bait-caught females. The level of lipid reserves in females coming to bait differs significantly between species: Ae.cantans has lipid reserves approximately double those of Ae.punctor . In addition, Ae.punctor is able to synthesize lipid to a level comparable with that found in bait-caught females after only 24 h, whilst it takes 192 h for Ae.cantans females to synthesize the amount of lipid found in host-seeking females, when allowed free access to sugar. Physiological differences in lipid synthesis and the level of lipid reserves required may therefore explain the differences observed between the species in the time taken to initiate host seeking.     

Suppression of methanogenesis by dissimilatory Fe(III)-reducing bacteria in tropical rain forest soils: implications for ecosystem methane flux

Tropical forests are an important source of atmospheric methane (CH₄), and recent work suggests that CH₄ fluxes from humid tropical environments are driven by variations in CH₄ production, rather than by bacterial CH₄ oxidation. Competition for acetate between methanogenic archaea and Fe(III)‐reducing bacteria is one of the principal controls on CH₄ flux in many Fe‐rich anoxic environments. Upland humid tropical forests are also abundant in Fe and are characterized by high organic matter inputs, steep soil oxygen (O₂) gradients, and fluctuating redox conditions, yielding concomitant methanogenesis and bacterial Fe(III) reduction. However, whether Fe(III)‐reducing bacteria coexist with methanogens or competitively suppress methanogenic acetate use in wet tropical soils is uncertain. To address this question, we conducted a process‐based laboratory experiment to determine if competition for acetate between methanogens and Fe(III)‐reducing bacteria influenced CH₄ production and C isotope composition in humid tropical forest soils. We collected soils from a poor to moderately drained upland rain forest and incubated them with combinations of ¹³C‐bicarbonate, ¹³C‐methyl labeled acetate (¹³CH₃COO^?), poorly crystalline Fe(III), or fluoroacetate. CH₄ production showed a greater proportional increase than Fe²⁺ production after competition for acetate was alleviated, suggesting that Fe(III)‐reducing bacteria were suppressing methanogenesis. Methanogenesis increased by approximately 67 times while Fe²⁺ production only doubled after the addition of ¹³CH₃COO^?. Large increases in both CH₄ and Fe²⁺ production also indicate that the two process were acetate limited, suggesting that acetate may be a key substrate for anoxic carbon (C) metabolism in humid tropical forest soils. C isotope analysis suggests that competition for acetate was not the only factor driving CH₄ production, as ¹³C partitioning did not vary significantly between ¹³CH₃COO^? and ¹³CH₃COO^?+Fe(III) treatments. This suggests that dissimilatory Fe(III)‐reduction suppressed both hydrogenotrophic and aceticlastic methanogenesis. These findings have implications for understanding the CH₄ biogeochemistry of highly weathered wet tropical soils, where CH₄ efflux is driven largely by CH₄ production.     

Soil organic matter dynamics during 80 years of reforestation of tropical pastures

Our research takes advantage of a historical trend in natural reforestation of abandoned tropical pastures to examine changes in soil carbon (C) during 80 years of secondary forest regrowth. We combined a chronosequence approach with differences in the natural abundance of ¹³C between C3 (forest) and C4 (pasture) plants to estimate turnover times of C in the bulk soil and in density fractions. Overall, gains in secondary forest C were compensated for by the loss of residual pasture-derived soil C, resulting in no net change in bulk soil C stocks down to 1 m depth over the chronosequence. The free light fraction (LF), representing physically unprotected particulate organic matter, was most sensitive to land-use change. Reforestation replenished C in the free LF that had been depleted during conversion to pastures. Turnover times varied with model choice, but in general, soil C cycling rates were rapid for the 0–10 cm depth, with even the heavy fraction (HF) containing C cycling in decadal time scales. Turnover times of C in the free LF from the 0–10 cm depth were shorter than for the occluded and HFs, highlighting the importance of physical location in the soil matrix for residence time in the soil. The majority of the soil C pool (82±21%) was recovered in the mineral-associated density fraction. Carbon-to-nitrogen ratios and differences in natural abundance ¹⁵N of soil organic matter (SOM) showed an increasing degree of decomposition across density fractions with increasing mineral association. Our data show that the physical distribution of C in the soil has a large impact on soil C turnover and the ability of soils to maintain SOM stocks during land-use and land-cover change.     

Silencing of EEF2K (eukaryotic elongation factor-2 kinase) reveals AMPK-ULK1-dependent autophagy in colon cancer cells

EEF2K (eukaryotic elongation factor-2 kinase), also known as Ca²⁺/calmodulin-dependent protein kinase III, functions in downregulating peptide chain elongation through inactivation of EEF2 (eukaryotic translation elongation factor 2). Currently, there is a limited amount of information on the promotion of autophagic survival by EEF2K in breast and glioblastoma cell lines. However, the precise role of EEF2K in carcinogenesis as well as the underlying mechanism involved is still poorly understood. In this study, contrary to the reported autophagy-promoting activity of EEF2K in certain cancer cells, EEF2K is shown to negatively regulate autophagy in human colon cancer cells as indicated by the increase of LC3-II levels, the accumulation of LC3 dots per cell, and the promotion of autophagic flux in EEF2K knockdown cells. EEF2K negatively regulates cell viability, clonogenicity, cell proliferation, and cell size in colon cancer cells. Autophagy induced by EEF2K silencing promotes cell survival and does not potentiate the anticancer efficacy of the AKT inhibitor MK-2206. In addition, autophagy induced by silencing of EEF2K is attributed to induction of protein synthesis and activation of the AMPK-ULK1 pathway, independent of the suppression of MTOR activity and ROS generation. Knockdown of AMPK or ULK1 significantly abrogates EEF2K silencing-induced increase of LC3-II levels, accumulation of LC3 dots per cell as well as cell proliferation in colon cancer cells. In conclusion, silencing of EEF2K promotes autophagic survival via activation of the AMPK-ULK1 pathway in colon cancer cells. This finding suggests that upregulation of EEF2K activity may constitute a novel approach for the treatment of human colon cancer.