Inflexibility of the plasma miRNA response following a high-carbohydrate meal in overweight insulin-resistant women.

CONTEXT: Metabolic inflexibility is a characteristic of insulin resistance, limiting the ability to transiently regulate oxidative metabolism and gene expression in response to nutrient availability. Little is known of the flexibility of post-transcriptional regulation, including circulatory miRNAs (c-miRNAs). DESIGN: The abundances of targeted c-miRNAs, with reported functions in metabolic regulation, were analysed in response to a high-carbohydrate meal in healthy weight insulin-sensitive (IS) and overweight insulin-resistant (IR) women. PARTICIPANTS: Age-matched healthy weight IS (n = 20, BMI = 24.3 ± 0.70) and overweight IR (n = 20, BMI = 28.6 ± 0.67) women. METHODS: An abundance of c-miRNAs was quantified prior to and following a high-carbohydrate breakfast meal (2500 kJ; 50% carbohydrate, 20% fat and 27% protein). Target genes of the differentially regulated c-miRNA were measured in RNA extracted from circulatory peripheral blood mononuclear cells (PBMCs). RESULTS: In healthy weight IS women, both miR-15a-5p (p = 0.03) and miR-17-5p (p < 0.01) levels were halved at 4 h post-meal. These miRNA remained unaltered following the same meal in the overweight IR women. Furthermore, amongst genes targeted by these miRNA, CPT1A (p = 0.01) and IL8 (p = 0.03) had also reduced expression 4 h post-meal only in the healthy weight IS women. CONCLUSIONS: The study findings provide preliminary evidence for a possible extension of metabolic inflexibility to include c-miRNAs. TRIAL REGISTRATION: The clinical trial is registered with Australian New Zealand Clinical Trials Registry under Trial registration: ANZCTR: ACTRN12615001108505. Registered on 21 October 2015.

The effects of vitamin D supplementation on muscle strength and mobility in postmenopausal women: a systematic review and meta-analysis of randomised controlled trials.

BACKGROUND: The results obtained from previous trials regarding the effects of vitamin D supplementation on muscle strength and mobility in postmenopausal women have been inconsistent. This systematic review and meta-analysis of randomised controlled trials (RCTs) aimed to investigate the effect of vitamin D supplementation on muscle strength and mobility in postmenopausal women. METHODS: A comprehensive search on EMBASE, PubMed, MEDLINE and SCOPUS was performed to identify relevant articles published up to 28 March 2019. RCTs published in English measuring the effect of all forms and doses of vitamin D supplementation with or without calcium on muscle strength and mobility outcomes in postmenopausal women were included. RESULTS: In total, 29 eligible studies were included in the systematic review. The pooled findings using a random effects model showed that vitamin D supplementation insignificantly increased hand grip strength (HGS) as the measurement of muscle strength (MD = 0.656; 95% confidence interval = -0.037 to 1.350, P = 0.06). However, it did not affect timed-up-and-go (TUG) as the measurement of mobility (MD = 0.118; 95% confidence interval = -0.655 to 0.892, P = 0.76). The subgroup analyses showed that vitamin D supplementation improved HGS with respect to dosages >1000 IU day-1 (P = 0.016), a treatment duration of 3 months (P Ë‚ 0.001) and subjects with baseline vitamin D <30 ng mL-1 (P = 0.033). CONCLUSIONS: The present review demonstrates that vitamin D supplementation resulted in small but nonsignificant improvements in muscle strength compared to control in postmenopausal women. No significant effect was observed in mobility after vitamin D administration.

Differences between bone mineral density, lean and fat mass of HIV-positive and HIV-negative black women

Objectives: To investigate the differences between bone mineral density (BMD), lean and fat mass of human immunodeficiency virus (HIV-) positive and HIV-negative black women and to investigate factors associated with low BMD. Methods: Case-control study of black women (n= 565) aged 29­65 years from Potchefstroom, North West province, South Africa, based on secondary analysis of data. Total BMD, left femur neck of the hip (LFN BMD), spine BMD, total fat, fat-free tissue mass and percentage body fat (%BF) were measured by dual-energy X-ray absorptiometry. Results: HIV-negative women had significantly higher median BMD, %BF, appendicular skeletal mass (ASM), ASM index, body mass index (BMI) and waist circumference than HIV-positive women. When the groups were matched for age and BMI, only spine BMD was marginally lower in HIV-positive women. In the total group, age, smoking and HIV status were associated with lower BMD, while calcium intake was positively associated with BMD. Similar variables were associated with BMD in HIV-negative women, while age and educational status were associated with BMD in HIV-positive women. Conclusion: Low BMD was more common among HIV-positive than HIV-negative women. Older HIV-positive women with low educational status are particularly at risk

Effects of Soy Phytoestrogens and New Zealand Functional Foods on Bone Health.

New Zealand is a rich source of food components that may have bioactivity on bone. Docosahexaenoic acid (DHA) from fish oil has been shown to maintain bone in ovariectomised (OVX) rats. Kiwifruit, a source of fibre and carotenoids, may also affect bone via a prebiotic as well as direct cell-based mechanisms. We aimed to 1) ascertain the effects of DHA on two cell models, including interactions with soy isoflavones; 2) and investigate the specific effects of carotenoids from kiwifruit as well as whole kiwifruit in cell-based and rodent models as well as in a human study. RAW 264.7 mouse monocytes or mouse bone marrow was used to generate osteoclasts (OC). Cells were exposed to the agents between 5 and 21 d and formation and activity of OC measured, including molecular markers. DHA inhibited OC formation in both cell models, including expression of cathepsin K, NFATc1 as well as actin ring formation. Combination with isoflavones enhanced these effects. In OVX rats and mice fed with kiwifruit for 8 wk, green kiwifruit reduced the rate of bone loss after OVX, and in mice it reduced C-telopeptide of Type 1 collagen (CTX) levels and RANKL expression while in menopausal women, green kiwifruit affected blood lipids and bone markers positively.

Diet, weight, cytokines and bone health in postmenopausal women.

OBJECTIVES: To investigate diet and nutrition-related factors associated with bone loss in a group of postmenopausal (PM) women. Nutritional intake, inflammatory markers and body composition (weight, body mass index, fat/lean mass) were analysed for associations with bone mineral density (BMD). DESIGN: A cross sectional study examining correlations between BMD (Duel-energy X ray absorptiometry; (DXA) and dietary intake (3-day diaries), body composition and plasma bone and inflammatory markers: C-terminal telopeptide of type I collagen (CTX) and procollagen type I N propeptide (P1NP), C- reactive protein (CRP), interleukin 6 and 10 (IL-6, IL-10), tumour necrosis factor (TNF) and osteoprotegerin (OPG). SETTING: Community dwelling women from the Auckland, Hawke's Bay and Manawatu regions in New Zealand. PARTICIPANTS: 142 healthy, PM women aged 50-70 years. RESULTS: OPG (per kilogram fat mass) was increased in women with osteoporosis (p<0.001) compared to groups classified with normal BMD and osteopenia. Protein, vitamin B12, zinc, potassium and dairy intake were all positively correlated with higher BMD while dairy and potassium intakes also inversely correlated with CTX. Body composition (weight, BMI and fat/lean mass) had strong positive associations with BMD. Multiple regression analysis showed body weight, potassium and dairy intake were predictors of increased BMD in PM women and explained 39% (r2=0.39, p< 0.003) of variance. CONCLUSION: BMD was negatively correlated with OPG and positively with weight, dairy and potassium intake. This study highlights the importance of maintaining adequate body weight and emphasising dairy and potassium predominantly sourced from fruit/vegetables to reduce bone loss at midlife.

The effect of aspirin and smoking on urinary excretion profiles of lactulose and mannitol in young women: toward a dynamic, aspirin augmented, test of gut mucosal permeability.

BACKGROUND: We explored the temporal dynamics of the lactulose mannitol test and the influence of a single dose of aspirin. METHODS: Twenty healthy female volunteers each received 600 mg aspirin or placebo in random sequence and were subsequently dosed with 10 g lactulose and 5 g mannitol, their urine collected every half hour for 6h. KEY RESULTS: The lactulose:mannitol ratios (LMR) of urine samples collected over the entire 6-h period were significantly higher than those collected in the first 3 h. Greater quantities of mannitol were excreted over the first than the subsequent 3 h. A similar pattern of temporal variation in mannitol excretion was found in smokers and non-smokers and was maintained following administration of a single 600 mg dose of aspirin. The rates at which lactulose was excreted were relatively constant over the entire 6 h period of collection, but mean levels were increased over the entire 6 h following the administration of aspirin. The effect of aspirin did not differ significantly between smokers and non-smokers. CONCLUSIONS & INFERENCES: While the LMR test is sufficiently sensitive to reproducibly detect the increase in intestinal permeability resulting from a single 600 mg oral dose of aspirin, the temporal patterns of excretion of mannitol and lactulose differ both in the presence and absence of aspirin. Hence, variation in sampling period and in method of dosage are likely to influence the result and it is preferable to examine the patterns of absorption of component sugars separately with due regard to the method of dosage.

High-calcium, vitamin D fortified milk is effective in improving bone turnover markers and vitamin D status in healthy postmenopausal Chinese women.

BACKGROUND/OBJECTIVES: Risk for developing osteoporosis increases in Asia. The purpose of the study was to evaluate the impact of a high-calcium vitamin D fortified milk (HCM) intervention on parathyroid hormone (PTH) levels, vitamin D status and markers of bone turnover in postmenopausal Chinese women. SUBJECTS/METHODS: Sixty three women (>55 years) were assigned to receive two servings of either a calcium/vitamin D fortified milk or a control drink for 12 weeks. PTH, serum 25 (OH)D levels, C-telopeptide of type I collagen (CTX) levels and procollagen type I N-terminal propeptide (PINP) were measured at baseline, 2, 8 and 12 weeks of supplementation. RESULTS: Daily calcium intake at baseline ranged between 260 and 482 mg for the HCM, and 252 and 692 mg for the control group. HCM improved serum 25 (OH)D levels significantly (33.13-39.49 nmol/l), while remaining similar in the control group (29.27-28.21 nmol/l). The difference between the groups were significant at week 2, 8 and 12. The percentage change in PTH levels in the HCM group was significant from week 2 onwards compared to the control drink (P<0.017, P<0.05 and P<0.001 at weeks 2, 8 and 12, respectively). Plasma CTX of the HCM group reduced by 25% between weeks 0 and 2, remaining significantly lower and at similar levels up to week 12. The difference between the HCM and control group for PINP reached significance at weeks 8 (P=0.011) and 12 (P=0.003). CONCLUSIONS: The HCM intervention significantly improved vitamin D status and reduced bone turnover over 12 weeks in postmenopausal Chinese women.

Long-chain polyunsaturated fatty acids: selected mechanisms of action on bone.

Evidence presented over the past 20 years has shown that long-chain polyunsaturated fatty acids (LCPUFAs), especially the n-3 fatty acids such as eicospentaenoic acid (EPA) and docosahexaenoic acid (DHA) are beneficial for bone health. Some studies in humans indicate that LCPUFAs can increase bone formation, affect peak bone mass in adolescents and reduce bone loss as measured using bone mineral densitometry. The cellular mechanisms of action of the LCPUFAs, however, are complex and involve modulation of fatty acid metabolites such as prostaglandins, resolvins and protectins, several signalling pathways, cytokines and growth factors. LCPUFAs affect receptor activator of nuclear factor κß (RANK), a receptor found on the osteoclast, the cell causing bone resorption, which controls osteoclast formation. Lipoxygenase (LOX) generated lipid mediators (resolvins, lipoxins, protectins and docosanoids) have both anti-inflammatory and pro-resolving activities. Both resolvins and lipoxins inhibit inflammation-induced bone resorption. Arachidonic acid significantly upregulates inducible NO synthase (iNOS) mRNA expression in human osteoblast-like cells, thereby possibly enhancing osteoclastic activity. The protective effect of EPA on osteoblastogenesis could be mediated by the biphasic cross-talk between PGE(2) and NO production involving COX-2 and iNOS pathways. Other mediators of osteoblast maturation include PPARα ligands such as linoleic acid and possibly DHA in association with bone morphogenic proteins. Since DHA is a weaker ligand for PPARγ, more uncommitted mesenchymal stem cells are thought to differentiate into osteoblasts rather than adipocytes. This review addresses selected cellular mechanisms that may explain the beneficial effects of the LCPUFAs on bone.

Vitamin D supplementation suppresses age-induced bone turnover in older women who are vitamin D deficient.

There is a lack of evidence that improving vitamin D status, without changing calcium intake, has a positive effect on bone turnover as indicated by bone marker changes. The objective was to measure the effect of vitamin D supplementation, in vitamin D deficient women (25(OH)D concentration<50 nmol/L), on osteocalcin (OC) and C-telopeptide (CTX). The study design was a randomised controlled intervention administering 4000 IU vitamin D3 or placebo daily for 6 months to South Asian women, aged>20 years. Subjects were stratified by age and menopausal status. Median (25th, 75th percentile) serum 25(OH)D increased significantly from 21 (11, 40) to 75 (55, 84) nmol/L with supplementation. In women>49 years or postmenopausal (n=26), who were not supplemented (n=13), CTX and OC levels increased (P=0.001, P=0.004 respectively), indicating an increased rate of bone turnover. With supplementation CTX decreased (P=0.012) and there was no significant change in OC. In women who were under 49 years and premenopausal (n=55; 29 supplemented), there was no significant response to supplementation in either CTX or OC. We conclude that correcting vitamin D deficiency in older women suppresses the age-induced increase in bone turnover and reduces bone resorption which would normally be exacerbated in conditions of low serum 25(OH)D.

Specific effects of gamma-linolenic, eicosapentaenoic, and docosahexaenoic ethyl esters on bone post-ovariectomy in rats.

Long chain polyunsaturated fatty acids (LCPUFAs) are involved in the regulation of bone metabolism. Increased dietary consumption of n-3, and possibly some n-6, LCPUFAs may limit postmenopausal bone loss. The aim of this study was to determine the effects on bone of specific fatty acids within the n-3 and n-6 LCPUFA families in ovariectomized (OVX) rats. Rats were OVX or sham-operated and fed either a control diet (OVX and sham) or a diet supplemented with 0.5 g/kg body weight/day of gamma-linolenic (GLA), eicosapentaenoic (EPA), docosahexaenoic (DHA) ethyl esters or a mixture of all three (MIX) for 16 weeks. Bone mineral content (BMC), area, and density and plasma concentrations of insulin-like growth factor-I, vitamin D, selected biochemical markers of bone metabolism, and parathyroid hormone (PTH) were determined. The OVX-induced decrease in lumbar spine BMC was significantly attenuated by DHA but not by EPA or GLA supplementation or supplementation with a mixture of all three LCPUFAs. Endosteal circumferences of tibiae were significantly greater in DHA and EPA compared to OVX. Plasma C-terminal telopeptide of type I collagen and osteocalcin concentrations were not significantly different in the DHA group compared to OVX. Femur BMC decreased by a significantly greater amount in GLA than OVX, and final plasma PTH concentrations were significantly higher in GLA compared to all other groups. In conclusion, DHA ameliorated OVX-induced bone mineral loss. GLA exacerbated post-OVX bone mineral loss, possibly as a result of PTH-induced bone catabolism.

Effects of arachidonic acid, docosahexaenoic acid and prostaglandin E(2) on cell proliferation and morphology of MG-63 and MC3T3-E1 osteoblast-like cells.

During bone remodelling bone is resorbed by osteoclasts and replaced again by osteoblasts through the process of bone formation. Clinical trials and in vivo animal studies suggest that specific polyunsaturated fatty acids (PUFAs) might benefit bone health. As the number of functional osteoblasts is important for bone formation the effects of specific PUFAs on in vitro osteoblastic cell proliferation were investigated. Morphological studies were conducted to determine whether exposure of the cells to these agents caused structural damage to the cells thereby yielding invalid results. Results from this study showed that arachidonic acid (AA) and docosahexaenoic acid (DHA) both inhibit cell growth significantly at high concentrations. The anti-mitotic effect of AA is possibly independent of PGE(2) production, as PGE(2) per se had little effect on proliferation. Further study is required to determine whether reduced proliferation due to fatty acids could be due to increased differentiation of osteoblasts to the mature mineralising osteoblastic phenotype.

Stimulation of prostaglandin E2 (PGE2) production by arachidonic acid, oestrogen and parathyroid hormone in MG-63 and MC3T3-E1 osteoblast-like cells.

Polyunsaturated fatty acids (PUFAs) as well as oestrogen (E2) and parathyroid hormone (PTH) affect bone cells. The aim of the study was to determine whether arachidonic acid (AA), E2, and PTH increase prostaglandin E(2) (PGE(2)) synthesis in MG-63 and MC3T3-E1 osteoblastic cells and the level of mediation by COX-1 and COX-2. PGE(2) levels were determined in the conditioned culture media of MG-63 and MC3T3-E1 osteoblasts after exposure to AA, PTH and E2. Cells were pre-incubated in some experiments with the unselective COX inhibitor indomethacin or the COX-2 specific blocker NS-398. Indirect immunofluorescence was performed on MG-63 cells to detect the presence and location of the two enzymes involved. AA increased PGE(2) secretion in both cell lines; production by MC3T3-E1 cells, however, was significantly higher than that of MG-63 cells. This could be due to autoamplification via the EP(1) subtype of PGE receptors in mouse MC3T3-E1 osteoblasts. Both COX-1 and COX-2 affected the regulation of PGE(2) synthesis in MG-63 cells. E2 had no effect on PGE(2) secretion in both cell lines, while PTH caused a slight increase in PGE(2) synthesis in the MG-63 cell line.

Changes in mineralization and biomechanics of tibial metaphyses in splinted rats.

The effect of 3 wk of splintage of a single hindlimb on the midarea and mineral content of both tibial metaphyses was assessed immediately after splint removal and after 1 mo of mobilization in 12-wk-old Sprague-Dawley rats. Immobilization reduced tibial metaphyseal bone mineral density (BMD) in immobilized limbs compared with "free" limbs of splinted animals and with controls. These changes persisted and were accentuated by relatively greater increases in tibial metaphyseal BMDs of unsplinted (control) animals after 7 wk. Immediately after splintage, tibial metaphyseal areas and total mineral contents of both hindlimbs of splinted animals were reduced compared with those of unsplinted animals. However, the relationship between mineralization and area differed between the free and immobilized limbs of splinted animals. The breaking strain and the breaking energy of immobilized and free femurs of splinted animals were impaired 4 wk after the removal of the splint. This impairment was correlated with an effect of splintage on femoral size with some additional local effect from immobilization. Thus osteoporotic changes consequent on immobilization include both local effects on mineralization and general effects on growth, which may separately influence the elastic properties of bone.

Intracellular free calcium in the neutrophils of maintenance haemodialysis patients.

Chronic renal failure has on occasion been referred to as a state of calcium toxicity. The aim of this study was to investigate the status of intracellular free Ca2+ in the neutrophils of chronic renal failure patients on maintenance haemodialysis treatment. Factors previously suggested to influence intracellular free Ca2+ were investigated including PTH levels, oxidative stress and recombinant human erythropoietin administration. The study involved 14 chronic renal failure patients on the haemodialysis programme of the Pretoria Academic hospital. Intracellular free Ca2+ and transmembrane Ca2+ fluxes were investigated by fluorescence spectrophotometry. Increases above control values were found in intracellular free Ca2+ (P-value 0.0242) and in the transmembrane Ca2+ flux upon fMLP stimulation (P-value 0.0002). The results showed significant differences in intracellular free Ca2+ between patients on rHuEPO and patients not on rHuEPO. The apparently rHuEPO-induced increase in intracellular free Ca2+ persisted in the presence of calcium channel blockers. No overt indications of oxidative stress could be detected by the antioxidant vitamin levels. It is concluded that factors other than those associated with uraemia, such as rHuEPO administration, might contribute to the often reported increase in intracellular free Ca2+ in these patients. Further studies to investigate the relationship between intracellular free Ca2+, rHuEPO and calcium channel blockers are suggested.

Red blood cell fatty acid profile of chronic renal failure patients receiving maintenance haemodialysis treatment.

The fatty acid profile of chronic renal failure (CRF) patients on maintenance haemodialysis treatment (MHT) is abnormal and results point towards an essential fatty acid (EFA) deficiency. However, controversies still exist as to which of the essential fatty acids (EFAs) or EFA-products are decreased. In this study, the results of a comprehensive analysis of the fatty acids, performed on the red blood cells of 14 CRF patients on MHT, are presented. The red blood cell membrane fatty acids determined in this study include a range of saturated fatty acids (SFAs), mono-unsaturated fatty acids (MUFAs) and poly-unsaturated fatty acids (PUFAs). Results confirmed the suggested presence of an essential fatty acid deficiency in CRF patients on MHT. It showed the total content of the n-6 fatty acids (31.66 +/- 3.21 vs 34.67 +/- 2.05), as well as the total content of the PUFAs (37.22 +/- 4.08 vs 40.93 +/- 2.35), to be significantly decreased. The total MUFA content was, in contrast, significantly increased (16.87 +/- 0.91 vs 15.49 +/- 1.18). The EFA deficiency profile seen in this study points towards that of a chronic inflammatory condition. This is borne out by the fact that all three precursors of the eicosanoids--the mediators of various inflammatory and immune responses--were reduced in the presence of an increase in MUFAs as well as SFAs. The possibility of the fatty acid profile of CRF patients on MHT being that of a chronic inflammatory condition is supported by the fact that continuous complement-dependent and complement-independent immune activation, due to bio-incompatibility between blood cells and the dialysis membranes, is known to occur during the dialysis process.

Effect of arachidonic acid on duodenal enterocyte ATPases.

Duodenal ion transport processes are supported by ATPase enzymes in basolateral membranes of the enterocyte. In vivo studies have shown that long term n-6 poly-unsaturated fatty acid (PUFA) supplementation in rats causes increases in intestinal Ca absorption, coupled with a higher total calcium balance and bone calcium content. The present in vitro study was undertaken to test the effect of arachidonic acid (AA), a highly unsaturated (and thus physiologically potent) member of the n-6 PUFA family, on ATPases in enterocyte basolateral membranes isolated with a sorbitol density gradient procedure. This paper presents results which show that AA inhibits Na+,K+-ATPase in a dose-dependent manner (-67% of basal activity at a concentration of 30 microg/ml, P < 0.005) but that this effect is not mediated by protein kinase C, as shown by the use of the protein kinase C blocker calphostin (0.5 microM). Indomethacin (IDM) at 0.1 mM, a cyclo-oxygenase blocker, could also not reverse the inhibitory effect of AA on Na+,K+-ATPase. Ca2+-ATPase, on the other hand, is not affected significantly (-10%, P > 0.05) by arachidonic acid at 30 microg/ml.

Upregulation of duodenal calcium absorption by poly-unsaturated fatty acids: events at the basolateral membrane.

Poly-unsaturated fatty acids, especially of the n-3 series, have a beneficial effect in treatment of osteoporosis in the elderly. Duodenal calcium absorption is a particularly vulnerable aspect of the development of this disease. It has been shown that the process of calcium transport through the rat duodenal enterocyte takes place in essentially three steps: entry of calcium through channels in the brush border (apical membrane), transcellular transport through the cytoplasm by calbindin and extrusion at the basolateral membrane by Ca(2+)-ATPase and a Ca(2+)-Na(+)exchanger which is driven by Na(+), K(+)-ATPase. This paper presents a hypothesis that poly-unsaturated fatty acids can modulate both Ca(2+)-ATPase and Na(+), K(+)-ATPase activity either by a direct action on the enzyme or by phosphorylation processes via protein kinases A and C and thus exert their positive influence on calcium absorption in this manner.

Modulation of intestinal vitamin D receptor availability and calcium ATPase activity by essential fatty acids.

The physiological mechanisms by which essential fatty acids (EFAs) affect calcium (Ca(2+)) retention is not clear, but suggestions have included changes in membrane fluidity, receptor modulation and induction of second messengers. The vitamin D receptor (VDR) is essential for the functioning of 1,25(OH)(2)D(3)which increases Ca(2+)absorption. Activity of the intestinal basolateral membrane (BLM) Ca(2+)ATPase correlates with the degree of Ca(2+)absorption. Therefore, changes in ATPase activity and VDR availability due to EFAs may influence calcium retention. We have investigated the effect of long-term dietary supplementation with EFAs on Ca(2+)ATPase activity (measured colourimetrically) and VDR availability (measured with the ELISA technique) after the loss of oestrogen induced by ovariectomy (OVX) in female Sprague Dawley rats. Control animals underwent anaesthesia and a surgical procedure but the ovaries were left intact (sham). Ca(2+)ATPase activity was significantly lower in OVX animals than in the intact animals (P<0.05) and following supplementation with EFAs, was significantly higher than in sham controls (P<0.05). A higher number of VDR was measured after OVX and declined due to EFA supplementation; these differences in activity of the ATPase and number of receptors could be ascribed to membrane changes due to EFA supplementation, feedback control by serum calcium or the direct influence of the EFAs.

Inhibition of duodenal enterocyte Mg2+-ATPase by arachidonic acid is not mediated by an effect on protein kinase C.

Active absorption processes in the duodenal enterocyte are driven by various ATPases. It is known that the activity of Na+,K+-ATPase, Ca2+-ATPase and Mg2+-ATPase can be modulated by polyunsaturated fatty acids of the n-6 series, for example by linoleic and gamma-linolenic acids. These effects may be achieved by protein phosphorylation via protein kinase C. The present study was undertaken to determine the effect of arachidonic acid on Mg2+-ATPase (measured colorimetrically) activity in basolateral membranes prepared from rat duodenum. It shows, for the first time, significant dose-dependent inhibition of Mg2+-ATPase (26-62%) by arachidonic acid (10-50 microg/ml) which already takes place after one minute of exposure, indicating involvement of a rapid signal transduction mechanism. Addition of the protein kinase C inhibitors bisimidolylmaleimide (2.5 microM) and calphostin (0.5 microM) did not influence the action of arachidonic acid on Mg2+-ATPase; protein kinase C involvement in this process is thus not indicated.