Changes In Membrane Fatty Acids Research Articles

Increasing temperature counteracts the negative effects of ultraviolet radiation on Microcystis aeruginosa under future climate scenarios in relation to physiological processes.

Heat waves, are a major concern related to climate change, and are projected to increase in frequency and severity. This temperature rise causes thermal stratification, exposing surface-dwelling organisms to higher levels of ultraviolet radiation (UVR). This study aims to understand how the toxic bloom-forming cyanobacterium Microcystis aeruginosa adapts to changing climatic conditions. The effects of increased temperature and UVR were evaluated in terms of cell abundance, reactive oxygen and nitrogen species (ROS/RNS), the antioxidant activity of catalase (CAT), superoxide dismutase (SOD), glutathione S transferase (GST), fatty acid (FA) content, and lipid damage. Negative UVR effects on biomass, lipid damage, and polyunsaturated fatty acids (PUFAs) were more pronounced at 26 °C compared to 29 °C. However, antioxidant responses were higher at 29 °C. The relative abundance of ω6 FAs was less affected by UVA, while ω3 FAs were highly sensitive at 29 °C but unsaturated fatty acids (UFA) did not experience peroxidation. The differential response in FA to high temperature and UVR results in differences in lipid damage and antioxidants. Changes in membrane FA may suggest an adaptation strategy at high UVR conditions. The exposure to environmental changes can alter membrane fluidity, affecting cell physiology. Thus, to survive UVR exposure, M. aeruginosa maintains a balance between damage and stress adaptation, increasing the protection of selected PUFAs at high temperatures, allowing them to effectively cope with the harmful effects of elevated temperature and UVR.

Direct Monitoring of Membrane Fatty Acid Changes and Effects on the Isoleucine/Valine Pathways in an ndgR Deletion Mutant of Streptomyces coelicolor.

NdgR, a global regulator in soil-dwelling and antibiotic-producing Streptomyces, is known to regulate branched-chain amino acid metabolism by binding to the upstream region of synthetic genes. However, its numerous and complex roles are not yet fully understood. To more fully reveal the function of NdgR, phospholipid fatty acid (PLFA) analysis with gas chromatography-mass spectrometry (GC-MS) was used to assess the effects of an ndgR deletion mutant of Streptomyces coelicolor. The deletion of ndgR was found to decrease the levels of isoleucine- and leucine-related fatty acids but increase those of valine-related fatty acids. Furthermore, the defects in leucine and isoleucine metabolism caused by the deletion impaired the growth of Streptomyces at low temperatures. Supplementation of leucine and isoleucine, however, could complement this defect under cold shock condition. NdgR was thus shown to be involved in the control of branched-chain amino acids and consequently affected the membrane fatty acid composition in Streptomyces. While isoleucine and valine could be synthesized by the same enzymes (IlvB/N, IlvC, IlvD, and IlvE), ndgR deletion did not affect them in the same way. This suggests that NdgR is involved in the upper isoleucine and valine pathways, or that its control over them differs in some respect.

Changes in membrane fatty acids of a halo-psychrophile exposed to magnesium perchlorate and low temperatures: Implications for Mars habitability

The presence of perchlorate salts in aqueous solutions bears two opposite effects on habitability. On the one hand, perchlorate salts trigger a decrease in the freezing point of the aqueous solutions, resulting in stable aqueous solutions at subzero temperatures, thereby widening the habitable conditions for potential microbial life. On the other hand, the presence of perchlorates in solution imposes a significant osmotic stress that compromises the integrity of microbial cell membranes, thereby restricting the habitable conditions in the same aqueous environment. Here we investigated the survivability and the changes in the composition of membrane fatty acids (FAs) of the bacterium Rhodococcus sp. JG-3 cells under warm (20°C), cold (4°C), and subzero temperatures (−10°C and −16°C), and in the presence (8 wt% and 16 wt%) and absence of magnesium perchlorate (Mg(ClO4)2). Bacterial cell survivability decreased with decreasing temperature and presence of magnesium perchlorate. However, Rhodococcus sp. JG-3 was able to tolerate up to 8 wt% Mg(ClO4)2 at −16°C. The presence of magnesium perchlorate in the medium decreased the concentration of total FAs, likely due to a destabilization of the molecules by the chaotropic effect of the perchlorate anion. At the maximum stress (both subzero temperatures and 16 wt% magnesium perchlorate), the composition of FAs changed, i.e., Rhodococcus sp. JG-3 cells increased the relative abundance of saturated FAs (SFAs) over the unsaturated (UFAs) or branched (BFAs). These changes in the proportion of FAs types may be a physiological response during cooling, aimed to improve lipid membrane stability. Interestingly, the composition and relative abundance of fatty acid types (i.e., SFAs, UFAs and BFAs) of Rhodococcus sp. JG-3 when simultaneously exposed to subzero temperatures and 16 wt% magnesium perchlorate was similar to that following freezing stress alone, suggesting that either both conditions triggered a similar response or that one response dominated over the other. Our findings contribute to understand the survivability and adaptation of extremophilic microorganisms under polyextreme conditions, such as those existing in the Martian subsurface today and/or in the past, which include the documented presence of magnesium perchlorate salts in ancient sediments and global cold temperatures.

Toxic Effect of Anionic Surfactants on Freshwater Sponge Lubomirskia baikalensis and Its Endosymbiotic Microalgae Chlorella sp.

A number of reports on sponge diseases, including from Lake Baikal, have increased dramatically all over the world in recent years. Herewith, there are various hypotheses for sponge mortality. Lubomirskia baikalensis (phylum Porifera, order Spongillida) is a unique endemic freshwater sponge of Lake Baikal that contains a complex community of eukaryotic and prokaryotic endosymbiotic microorganisms. In this work, we present the first results for the effect of anionic surfactants viz. linear alkylbenzene sulphonates (LAS) at low 10 and 20 µg L−1 concentrations on Baikal sponge species and their symbiotic community as an experimental model. A new toxicity test protocol under conditions close to natural is proposed. It uses the sponge amoebocytes called as SA1-cells, which contain eukaryotic green microalgae Chlorella sp. These SA1-cells are shown to be representative indicator in assessing the impact of anionic surfactants. The acute toxic effect resulted in 97–100% sponge cell death in less than 48 h, as well as 100% symbiotic microalgae Chlorella sp. death over 72 h was noted under LAS solution (20 μg L−1) exposure during in vivo experiments. This includes the cell membrane fatty acid changes, change in the cell sizes, cell swelling, and lysis. Long term exposure to LAS solution (10 μg L−1) reflected in cellular stress (oxidative stress) and accompanied by malondialdehyde formation (0.16–2.0 μg g−1 of dry weight) during 14-day exposure was noted. Oxidative stress and mortality of L. baikalensis are associated with their low antioxidant activity. Trolox-equivalent antioxidant capacity (TEAC) found in the range from 0.00031 to 0.00077 Trolox equivalents for these freshwater sponges.

Cold stress-induced changes in metabolism of carbonyl compounds and membrane fatty acid composition in chickpea

In this study, changes in membrane fatty acid (FA) composition and damage indices contents as well as the transcript patterns of carbonyl-detoxifying genes were evaluated in two chickpea (Cicer arietinum L.) genotypes, cold-tolerant Sel96th11439 and cold-sensitive ILC533 under cold stress (CS; 4 °C). During CS, H2O2 and malondialdehyde (MDA) contents increased (by 47% and 57%, respectively) in the sensitive genotype, while these contents remained unchanged in the tolerant genotype. In tolerant plants, higher content of linoleic, linolenic, unsaturated FAs (UFAs), total FAs and double bond index (DBI) (by 23, 21, 19, 17 and 9%, respectively) was observed at 6 days after stress (DAS) compared to sensitive plants, which, along with alterations of the damage indices, indicate their enhanced tolerance to CS. Compared with the sensitive genotype, less lipoxygenase (LOX) activity (by 59%) in the tolerant genotype was accompanied by decreased MDA and increased levels of UFAs and DBI during CS, particularly at 6 DAS. Upregulation of aldehyde dehydrogenase and aldo-keto reductase genes (by 9- and 10-fold, respectively) at 1 DAS, along with the enhanced transcript levels of aldehyde reductase and 2-alkenal reductase (by 3- and 14.7-fold, respectively) at 6 DAS were accompanied by increased UFAs and reduced MDA contents in the tolerant genotype. Overall, the results suggest that cold tolerance in chickpea was partly associated with regulation of membrane FA compositions and the potential metabolic networks involved in synthesis and degradation of carbonyl compounds.

P0326EFFECT OF PRAVASTATIN ON ERYTHROCYTE MEMBRANE FATTY ACID CONTENTS IN PATIENTS WITH CHRONIC KIDNEY DISEASE

Abstract Background and Aims Treatment with statin has been decreased the risk of cardiovascular events in patients with chronic kidney disease (CKD). Erythrocyte membrane oleic acid contents are significantly higher in patients with acute coronary syndrome than control subjects. Still, there is no report about the effect of statin on erythrocyte membrane fatty acid (FA) including oleic acid. The aim of this study was to evaluate the effect of pravastatin on the erythrocyte membrane FA in patients with CKD. Method Sixty two CKD patients were enrolled in this single arm randomized clinical trial and this trial was conducted at two centers from Jan 2017 to March 2019 (NCT02992548). Pravastatin with dose of 20mg was initially treated for 24 weeks. Pravastatin dose was increased to 40mg after 12 weeks, if it is necessary to control dyslipidemia. The primary outcome of this study was the change of erythrocyte membrane FA including oleic acid after pravastatin treatment for 24 weeks. We checked total cholesterol, triglyceride, LDL-cholesterol, HDL-cholesterol and adiponectin for secondary outcome at baseline and after 24 weeks. Erythrocyte membrane FA contents were measured by gas chromatography at baseline and after 24 weeks. Results Forty-five patients finished this study (age: 59.2±12.4 years, male: 42.2%, diabetes: 48.9%). There was no adverse effect related with pravastatin. Baseline serum creatinine was 1.5±0.7 mg/dL and estimated glomerular filtration rate was 54.2±27.3 ml/min/1.73 m2. Compared to baseline, total cholesterol (223.1±50.8 mg/dL vs. 168.4±32.5 mg/dL), LDL-cholesterol (149.1±35.3mg/dL vs. 100.1±25.4 mg/dL) and C-reactive protein were significantly decreased after pravastatin treatment. There were no significant changes in triglyceride, HDL-cholesterol, serum creatinine, amounts of proteinuria and adiponectin levels. Saturated FA, oleic acid and arachidonic acid contents of erythrocyte membrane were significantly increased after pravastatin treatment compared to baseline levels. Polyunsaturated FA (PUFA) and linoleic acid were significantly decreased after pravastatin treatment compared to baseline. Decreased tendency of eicosapentaenoic acid and omega-3 index were found after pravastatin treatment. Conclusion Linoleic acid or omega-3 FA supplementations may be necessary to recover erythrocyte membrane FA changes, when pravastatin is used for dyslipidemia treatment in CKD patients. Further study for reducing cardiovascular events are necessary by using combined PUFA and pravastatin treatment.

Effect of silver nanoparticles on gill membranes of common carp: Modification of fatty acid profile, lipid peroxidation and membrane fluidity

Although the toxicity of silver nanoparticles (AgNPs) in aquatic organisms has been extensively investigated, the mechanism by which AgNPs damage membranes remains unclear. This study investigated the toxic effects of a series of sub-lethal concentrations of AgNPs on the membranes of freshwater carp (Cyprinus carpio) gills, based on changes in membrane fatty acid (FA) profile, membrane fluidity, membrane lipid peroxidation, and histopathology. Most of the FAs in fish gill membrane was not significantly affected by exposure to multiple AgNPs concentrations, only few significant changes occurred in some specific FAs species at a high concentration of AgNPs exposure. In particular, high concentrations of AgNPs significantly decreased the proportions of two important long-chain n-3 series polyunsaturated FAs (C20: 5n3, and C22: 6n3), resulting in a decreased ratio of n-3 polyunsaturated FAs to n-6 polyunsaturated FAs (Σn-3UFA/Σn-6UFA). The AgNPs also caused a dose-dependent decrease in fish gill membrane fluidity, increased the level of lipid peroxidation, and inhibited Na+/K+-ATPase enzyme activity. Further histopathological examination revealed that exposure to AgNPs can cause toxic responses in the lamellae, including the thinning of the basement membrane, malformation, and inflammation. Together, the results suggest that the mechanism of AgNPs membrane toxicity involves the oxidization of long-chain omega-3 unsaturated FAs to saturated FAs via lipid peroxidation, resulting in, decreased membrane fluidity and ultimately the destruction of the normal physiological function of the fish gill membrane. The findings contribute significantly to our understanding of nanoparticle-induced membrane toxicity and potential risks in aquatic environments.

Effects of somatostatin, curcumin, and quercetin on the fatty acid profile of breast cancer cell membranes.

Breast cancer is a worldwide commonly found malignancy in women and effective treatment is regarded as a huge clinical challenge even in the presence of several treatment options. Extensive literature is available demonstrating polyphenols as phytopharmaceutical anticancer agents. Among the polyphenols, quercetin and curcumin have been reported to have a strong potential against breast cancer. However, so far, no comprehensive study has been performed to demonstrate the anticarcinogenic effects of curcumin, quercetin, and their combinations with somatostatin on the fatty acid profile of breast cancer cell membranes. We used MCF-7 and MDA-MB231 breast cancer cells incubated with curcumin and quercetin for 24 h, in the absence and presence of somatostatin, at their EC50 concentrations to evaluate membrane fatty acid based functional lipidomics together with the followup of EGFR and MAPK signaling pathways. The two cell lines gave different membrane free fatty acid reorganization. In MCF-7 cells, the following changes were observed: an increase of ω6 linoleic acid in the cells incubated with somatostatin + quercetin and quercetin and a decrease of ω3 acids in the cells incubated with somatostatin + curcumin compared to somatostatin and significant increases of monounsaturated fatty acid (MUFA), mono-trans arachidonic acid levels and docosapentaenoic acid for the cells incubated with somatostatin + quercetin compared to the control cells. In MDA-MB231 cells, incubations with curcumin, quercetin, and somatostatin + quercetin induced the most significant membrane remodeling with the increase of stearic acid, diminution of ω6 linoleic, arachidonic acids, and ω3 (docosapentaenoic and docosahexaenoic acids). Distinct signaling pathway changes were found for these cell lines. In MCF-7 cells, separate or combined incubations with somatostatin and quercetin, significantly decreased EGFR and incubation with curcumin decreased MAPK signaling. In MDA-MB231 cells, incubation with curcumin decreased AKT1 and p-AKT1 (Thr308) levels. Incubation with curcumin and quercetin decreased the EGFR levels. Our results showed that cytostatic and antioxidant treatments can be combined to induce membrane fatty acid changes, including lipid isomerization as specific free radical driven process, and to influence signaling pathways. This study aimed to contribute to the literature on these antioxidants in the treatment of breast cancer to clarify the effects and mechanisms in combination with somatostatin.

Resistance of Escherichia coli O157:H7 ATCC 35150 to ohmic heating as influenced by growth temperature and sodium chloride concentration in salsa

Salsa is a liquid-solid food containing jalapeño and serrano peppers, which result in multistate outbreak in 2008. Storage temperature and sodium chloride (NaCl) concentration of salsa vary depending on the climate, season, and type of product. In this regard, effect of growth conditions, namely, low temperature (15 °C) or NaCl concentration (4.5%) on the resistance of Escherichia coli O157:H7 ATCC 35150 to ohmic heating was identified in this study. Cells of E. coli O157:H7 ATCC 35150 grown under different growth conditions was inoculated into prepared salsa sample, and then subjected to ohmic heating. Mechanisms of resistance acquisition were identified by transcriptional responses, membrane fatty acid changes and confirmed with propidium iodide (PI) uptake values. Resistance of the pathogen to ohmic heating decreased when growth temperature decreased from 37 °C to 15 °C while increased resistance was observed for this pathogen when grown with 4.5% NaCl. Several heat stress related genes such as dnaK, rpoH, grpE, groES, htpG, and htpX were up-regulated (≥5 fold change) as growth temperature decreasedwhile groEL, dnaK, rpoH were up-regulated when grown with high NaCl concentration in the present study. The ratio of unsaturated fatty acids (USFA) to saturated fatty acids (SFA) of pathogen increased slightly (+0.16) or significantly (+0.79) with increasing NaCl concentration or decreasing temperature, respectively. These results indicate that the cell membrane of the pathogen grown at low temperature was more susceptible to heat than when grown under optimal conditions or high NaCl concentration. Cell membrane damage measured by PI uptake values of the pathogen grown with high NaCl concentration were not significantly different from those of the control (p > 0.05), while the values were significantly higher for the pathogen grown at low temperature and subjected to ohmic heating (p < 0.05). Based on these results we suggest that resistance of the pathogen grown at low temperature to ohmic heating decreased because of dominant cell membrane damage compared to induced heat stress related genes. The cell membrane damage was dominant by means of an increased ratio of USFA to SFA. On the other hand, pathogen resistance increased when grown in medium of high NaCl concentration because of induced heat stress related genes.

Effects of Somatostatin and Vitamin C on the Fatty Acid Profile of Breast Cancer Cell Membranes.

Vitamin C (Vit C) is an important physiological antioxidant with growing applications in cancer. Somatostatin (SST) is a natural peptide with growth inhibitory effect in several mammary cancer models. The combined effects of SST and Vit C supplementation have never been studied in breast cancer cells so far. We used MCF-7 and MDA-MB231 breast cancer cells incubated with SST for 24h, in the absence and presence of Vit C, at their EC50 concentrations, to evaluate membrane fatty acid-profiles together with the follow-up of EGFR and MAPK signaling pathways. The two cell lines gave different membrane reorganization: in MCF-7 cells, decrease of omega-6 linoleic acid and increase of omega-3 fatty acids (Fas) occurred after SST and SST+Vit C incubations, the latter also showing significant increases in MUFA, docosapentaenoic acid and mono-trans arachidonic acid levels. In MDA-MB231 cells, SST+Vit C incubation induced significant membrane remodeling with an increase of stearic acid and mono-trans-linoleic acid isomer, diminution of omega-6 linoleic, arachidonic acid and omega-3 (docosapentaenoic and docosadienoic acids). Distinct signaling pathways in these cell lines were studied: in MCF-7 cells, incubations with SST and Vit C, alone or in combination significantly decreased EGFR and MAPK signaling, whereas in MDA-MB231 cells, SST and Vit C incubations, alone or combined, decreased p- P44/42 MAPK levels, and increased EGFR levels. Our results showed that SST and Vit C can be combined to induce membrane fatty acid changes, including lipid isomerization through a specific free radical-driven process, influencing signaling pathways.

Exogenous Fatty Acids Protect Enterococcus faecalis from Daptomycin-Induced Membrane Stress Independently of the Response Regulator LiaR.

Enterococcus faecalis is a commensal bacterium of the gastrointestinal tract that can cause nosocomial infections in immunocompromised humans. The hallmarks of this organism are its ability to survive in a variety of stressful habitats and, in particular, its ability to withstand membrane damage. One strategy used by E. faecalis to protect itself from membrane-damaging agents, including the antibiotic daptomycin, involves incorporation of exogenous fatty acids from bile or serum into the cell membrane. Additionally, the response regulator LiaR (a member of the LiaFSR [lipid II-interacting antibiotic response regulator and sensor] system associated with cell envelope stress responses) is required for the basal level of resistance E. faecalis has to daptomycin-induced membrane damage. This study aimed to determine if membrane fatty acid changes could provide protection against membrane stressors in a LiaR-deficient strain of E. faecalis We noted that despite the loss of LiaR, the organism readily incorporated exogenous fatty acids into its membrane, and indeed growth in the presence of exogenous fatty acids increased the survival of LiaR-deficient cells when challenged with a variety of membrane stressors, including daptomycin. Combined, our results suggest that E. faecalis can utilize both LiaR-dependent and -independent mechanisms to protect itself from membrane damage. Enterococcus faecalis is responsible for a significant number of nosocomial infections. Worse, many of the antibiotics used to treat E. faecalis infection are no longer effective, as this organism has developed resistance to them. The drug daptomycin has been successfully used to treat some of these resistant strains; however, daptomycin-resistant isolates have been identified in hospitals. Many daptomycin-resistant isolates are found to harbor mutations in the genetic locus liaFSR, which is involved in membrane stress responses. Another mechanism shown to increase tolerance to daptomycin involves the incorporation of exogenous fatty acids from host fluids like serum or bile. This improved tolerance was found to be independent of liaFSR and suggests that there are additional ways to impact sensitivity to daptomycin. Thus, further studies are needed to understand how host fatty acid sources can influence antibiotic susceptibility.

Cell membrane fatty acid changes and desaturase expression of Saccharomyces bayanus exposed to high pressure homogenization in relation to the supplementation of exogenous unsaturated fatty acids.

Aims: The aim of this work was to study the responses of Saccharomyces bayanus cells exposed to sub-lethal high-pressure homogenization (HPH) and determine whether the plasmatic membrane can sense HPH in the presence, or absence, of exogenous unsaturated fatty acids (UFAs) in the growth medium.Methods and Results: High-pressure homogenization damaged and caused the collapse of cell walls and membranes of a portion of cells; however, HPH did not significantly affect S. bayanus cell viability (less than 0.3 Log CFU ml-1). HPH strongly affected the membrane fatty acid (FA) composition by increasing the percentage of total UFA when compared with saturated fatty acids. The gene expression showed that the transcription of OLE1, ERG3, and ERG11 increased after HPH. The presence of exogenous UFA abolished HPH-induced effects on the OLE1 and ERG3 genes, increased the percentage of membrane lipids and decreased the expression of OLE1 and ERG3 within 30 min of treatment.Conclusion: The results suggest a key role for UFA in the microbial cell response to sub-lethal stress. In addition, these data provide insight into the molecular basis of the response of S. bayanus to this innovative technology.Significance and Impact of the Study: Elucidation of the mechanism of action for sub-lethal HPH will enable the utilization of this technology to modulate the starter performance at the industrial scale.

Studies on the Mechanism of Telavancin Decreased Susceptibility in a Laboratory-Derived Mutant

Telavancin is a novel semisynthetic lipoglycopeptide derivative of vancomycin with a dual mode of action. This study sought to understand the mechanisms of decreased telavancin susceptibility in a laboratory-derived Staphlococcus aureus mutant Tlv(DS)MED1952. There were extensive changes in the transcriptome of Tlv(DS)MED1952 compared to the susceptible parent strain MED1951. Genes upregulated included cofactor biosynthesis genes, cell wall-related genes, fatty acid biosynthesis genes, and stress genes. Downregulated genes included lysine operon biosynthesis genes and lrgB, which are induced by telavancin in susceptible strains, agr and kdpDE genes, various cell surface protein genes, phenol-soluble modulin genes, several protease genes, and genes involved in anaerobic metabolism. The decreased susceptibility mutant had somewhat thicker cell walls and a decreased autolytic activity that may be related to decreased proteolytic peptidoglycan hydrolase processing. Membrane fatty acid changes correlated with increased membrane fluidity were observed. It seems likely that there are multiple genetic changes associated with the development of decreased telavancin susceptibility. The Tlv(DS) mutant showed some similar features to vancomycin-intermediate S. aureus and decreased daptomycin susceptibility strains, but also exhibited its own unique features.

Changes in membrane fatty acids and murein composition of Bacillus cereus and Salmonella Typhi induced by gamma irradiation treatment

This present study was carried out to evaluate the effects of gamma irradiation on the fatty acids (FA) and mureins composition of two different radiotolerant bacteria. Bacillus cereus LSPQ 2872 and Salmonella Typhi ATCC 19430 were used for this study. The bacterial strains were treated with a sublethal radiation dose of 1 kGy to cause a cellular damage. Gas chromatography (GC) and high performance liquid chromatography (HPLC) analysis were performed to demonstrate respectively the modification of the FA composition and the changes in muropeptide profile. Results obtained show, for both bacteria, that this treatment had a significant effect ( P ≤ 0.05) on the FA content with an increase of unsaturated FA percentage. Substantial changes were also noticed for the relative percentage and the number of the muropeptides. This study represents one of the few to demonstrate the modifications on bacterial membrane as a cellular response to survive the ionising radiation stress.

Consumption of Virgin Olive Oil Influences Membrane Lipid Composition and Regulates Intracellular Signaling in Elderly Adults With Type 2 Diabetes Mellitus

We aimed to define changes in membrane fatty acids and signaling proteins induced by virgin olive oil (VOO) consumption in elderly persons with type 2 diabetes (n = 16) compared to a control group (n = 28). The fatty acid composition was determined by gas chromatography and G-protein subunits and protein kinase C alpha (PKCalpha) by immunoblotting. VOO consumption increased the monounsaturated fatty acid content in phospholipids and cholesterol esters in both groups. In contrast, saturated fatty acids were decreased only in phospholipids. The levels of Galphao, Gbeta, and PKCalpha were significantly lower in diabetics than in controls. However, whereas VOO consumption reduced Galphas, Gbeta, and PKCalpha in both groups, reduction in Galphai was observed only in diabetics. These results indicate that long-term VOO consumption modifies the fatty acid composition of plasma membrane, which influences the association of G proteins and PKCalpha with the lipid bilayer. These combined effects probably account for the positive effects of VOO on glycemic homeostasis.

Drought acclimation confers cold tolerance in the soil collembolan Folsomia candida

It has been noted that both summer drought and sub-zero winter temperatures induce the synthesis of sugars and polyols in invertebrate tissues. This has led several authors to suggest that many of the adaptations, previously viewed as a response to cold, might be part of a more universal desiccation tolerance mechanism. Here we show that acclimation of the soil dwelling collembolan Folsomia candida to a sublethal desiccation stress confers tolerance to cold shock and a significant increase in the molar percent of membrane fatty acids with a mid-chain double bond. These changes in membrane fatty acids are interpreted as conferring a significant reduction in the transition temperature of cell membranes, as would be expected in acclimation to cold, and these changes are therefore interpreted as contributing to the cross-tolerance. Drought acclimation was also shown to trigger the synthesis of the 70 kDa family of heat-shock proteins (Hsp70). This group of heat shock proteins is implicated in the reestablishment of the normal three-dimensional structure of partially unfolded proteins and therefore are also likely to contribute to the observed cross-tolerance. This study provides evidence that the stresses exerted by desiccation and cold at the cellular level have sufficient similarities to induce overlapping adaptations.

Cold shock and its effect on ribosomes and thermal tolerance in Listeria monocytogenes.

Differential scanning calorimetry (DSC) and fatty acid analysis were used to determine how cold shocking reduces the thermal stability of Listeria monocytogenes. Additionally, antibiotics that can elicit production of cold or heat shock proteins were used to determine the effect of translation blockage on ribosome thermal stability. Fatty acid profiles showed no significant variations as a result of cold shock, indicating that changes in membrane fatty acids were not responsible for the cold shock-induced reduction in thermal tolerance. Following a 3-h cold shock from 37 to 0 degrees C, the maximum denaturation temperature of the 50S ribosomal subunit and 70S ribosomal particle peak was reduced from 73.4 +/- 0.1 degrees C (mean +/- standard deviation) to 72.1 +/- 0.5 degrees C (P < or = 0.05), indicating that cold shock induced instability in the associated ribosome structure. The maximum denaturation temperature of the 30S ribosomal subunit peak did not show a significant shift in temperature (from 67.5 +/- 0.4 degrees C to 66.8 +/- 0.5 degrees C) as a result of cold shock, suggesting that either 50S subunit or 70S particle sensitivity was responsible for the intact ribosome fragility. Antibiotics that elicited changes in maximum denaturation temperature in ribosomal components also elicited reductions in thermotolerance. Together, these data suggest that ribosomal changes resulting from cold shock may be responsible for the decrease in D value observed when L. monocytogenes is cold shocked.

Changes in membrane fatty acids and delta-9 desaturase in senescence accelerated (SAMP8) mouse hippocampus with aging

Senescence accelerated mice (SAMP8) exhibit age induced impairments such as loss of memory and learning disabilities by the age of 8–10 months. Analysis of hippocampus of SAMP8 mice revealed that delta 9-desaturase ( Δ 9desaturase) activity reduced up to 44–50% with age. Correspondingly, levels of unsaturated fatty acids are also lowered in the aged animals approximately to the same levels. Rnase protection assay showed that Δ 9specific message decreased similarly with age. As such a decrease is known to cause alterations in membrane fluidity and affect cellular signaling pathways, these results suggest that lowering of Δ 9gene expression may be partly involved in age induced impairments.

Changes in Membrane Fatty Acids in Cold-acclimated Turfgrass

Three genotypes of seashore paspalum, `PI 299042', `Adalayd', and `PI 509018-1' considered to be cold-sensitive, intermediately cold-tolerant and cold-tolerant, respectively, were analyzed to investigate the biochemical basis of cold tolerance. The cultivars were acclimated to 8/4 °C day/night temperatures and rhizomes nodes and crowns were harvested at 7-day intervals over the 4-week experiment. Total lipid was extracted from these tissues, and the fatty acids present in the lipid fraction were identified by gas chromatography. Palmitic acid, stearic acid, linoleic acid and linolenic acid were the major fatty acids present. In cold acclimated tissues, the level of palmitic acid and stearic acid did not change significantly during the treatment period. There was a decrease in the level of linoleic acid by the second week of cold treatment. The amount of linolenic acid increased significantly during the second week of cold treatment corresponding to the decrease in linoleic acid. The change in the amount was significantly greater in `PI 509018-1' than in `Adalayd' or `PI 299042'. These results are similar to what was found for cultivars of bermudagrass that differ in their cold-tolerance phenotypes. Desaturases are enzymes involved in introducing the double bonds into the fatty acid chains. Research is underway to characterize and clone the genes encoding the Ω3, Ω6, and Ω9 desaturases, which may have an important role in affecting the cold tolerance by altering the degree of membrane lipid fatty acid saturation.

Increased mean platelet volume after oestrogen replacement therapy.

The effect of hormone replacement therapy (HRT) on platelet size was examined in 38 post-menopausal women before and at end of a 6 week period of HRT. Subjects were treated with cyclical L-norgestrel 75 mg daily from days 17 to 28 of a 28-day cycle combined with continuous conjugated equine oestrogens 0.625 mg daily. Platelet-rich plasma was obtained to measure platelet indices by Coulter analysis. Plasma measurements of luteinizing hormone, follicle stimulating hormone and 17 beta-oestradiol were measured by immunoassays. Platelet membrane fatty acids were measured by gas-liquid chromatography. A significant reduction in platelet membrane linoleic acid, di-homo-gamma-linoleic acid and arachidonic acid of 8.1%, 14.3% and 17.8%, respectively was noted after 6 weeks of HRT (p < 0.01). There was an increase in the platelet count (not significant) and platelet volume (MPV) (P < 0.05) after 6 weeks of hormone therapy. HRT appears to increase mean platelet volume which may indicate an increase in platelet reactivity. There was no correlation between the changes in mean platelet volume and membrane fatty acid changes in platelets after such therapy.