Milk fat globule membrane and its polar lipids: reviewing preclinical and clinical trials on cognition.

In most parts of the world, life expectancy is increasing thanks to improved healthcare, public health policies, nutrition, and treatment. This increase in lifespan is often not accompanied by an increase in health span, which severely affects people as they age. One notable consequence of this is the increasing prevalence of neurodegenerative diseases such as mild cognitive impairment, dementia, and Alzheimer's disease. Therefore, dietary and pharmaceutical measures must be taken to reduce the burden of such pathologies. Among the different types of nutrients found in the diet, lipids and especially polar lipids are very important for cognition due to their abundance in the brain. Amid the most studied sources of polar lipids, milk fat globule membrane (MFGM) stands out as it is abundant in industrial by-products such as buttermilk. In this narrative review, we discuss the latest, i.e. less than five years old, scientific evidence on the use of MFGM and its polar lipids in cognitive neurodevelopment in early life and their potential effect in preventing neurodegeneration in old age. We conclude that MFGM is an interesting, abundant and exploitable source of relatively inexpensive bioactive molecules that could be properly formulated and utilized in the areas of neurodevelopment and cognitive decline. Sufficiently large randomized controlled trials are required before health-related statements can be made. However, research in this area is progressing rapidly and the evidence gathered points to biological, health-promoting effects.

Associations of Dietary Intake and Nutrient Status with Micronutrient and Lipid Composition in Breast Milk of Donor Women.

The influence of the diet and nutritional status of milk donors on the nutritional composition of donor human milk (DHM) is unknown. The present study aimed to determine the nutritional profile of DHM and the associations between donors' dietary intake and nutritional status and the micronutrient and lipid composition in DHM. For this purpose, 113 donors completed a food frequency questionnaire, provided a five-day weighed dietary record, and collected milk for five consecutive days. Nutrient determinations in donors' erythrocytes, plasma, urine, and milk were performed. Multiple linear regressions were conducted for the evaluation of the associations. We highlight the following results: DHM docosahexaenoic acid (DHA) was positively associated with donors' plasma DHA content and donors' DHA intake (R2 0.45, p < 0.001). For every 1 g/day DHA intake, an increase of 0.38% in DHA content and 0.78% in total omega-3 content was observed in DHM (R2 0.29, p < 0.001). DHM saturated fatty acids were positively associated with erythrocyte dimethyl acetals, plasma stearic acid, trans fatty acids intake, and breastfeeding duration and negatively associated with erythrocyte margaroleic acid (R2 0.34, p < 0.01). DHM cholecalciferol was associated with plasma cholecalciferol levels and dairy intake (R2 0.57, p < 0.01). Other weaker associations were found for free thiamin, free riboflavin, pyridoxal, dehydroascorbic acid, and the lipid profile in DHM. In conclusion, the diet and nutritional status of donors influence the fatty acid profile and micronutrient content of DHM.

Reduction of Beta Cyclodextrin by Curd Washing in Low-Cholesterol Manchego Cheese.

Beta-cyclodextrin (ß-CD) is a cyclic oligosaccharide consisting of seven glucose units. ß-CD is increasingly used in food research to reduce cholesterol due to its affinity for non-polar molecules such as cholesterol and as a natural additive. The purpose of this study was to evaluate the effect of curd washing in ewe's milk cheese on the reduction in cholesterol by ß-CD from pasteurized ewe's milk Manchego cheese and the characteristics of its main components: milk, lipids, and flavor. An approximately 98.45% cholesterol reduction was observed in washed experimental cheeses that were treated by using ß-CD. The remaining residual ß-CD from the effect of curd washing was 0.15% in mature cheese, of the initial 1% ß-CD treatment of the milk. The chemical properties (fat, moisture, and protein) did not change as a result of the curd washing with or without ß-CD. The curd washing with or without ß-CD on the levels of the various lipid fraction (fatty acids, triglycerides, and phospholipids) were comparable in treated and untreated cheeses. The effects of curd washing and the ß-CD treatment did not significantly affect flavor components or short chain free fatty acids. The ß-CD molecules were edible and nontoxic; as a result, they could be used safely in cholesterol removal processing in cheese manufacturing, improving the reduction in residual ß-CD by curd washing by 85%. Therefore, the present study suggests that curd washing combined with ß-CD is an effective process for cholesterol removal in Manchego cheese, preserving its desirable properties.

Food matrix impacts bioaccessibility and assimilation of acid whey-derived milk fat globule membrane lipids in Caco-2 cells.

The milk fat globule membrane (MFGM) imparts human health benefits ranging from improved immune system, gut, and brain function to improved cardiometabolic health. The industry's growing interest in introducing MFGM-enriched foods requires scientific evidence that the benefits derived from this compound are not affected by the formulation or processes that may alter its function, such as the digestion process. In this study, the impact of food matrices and supplementation levels on the bioaccessibility and assimilation of MFGM lipids in cell culture was investigated. Three food matrices including a protein-rich jelly, carbohydrate-rich cookie, and a carbohydrate- and fat-rich cookie with sunflower oil (SF-cookie) were supplemented with an MFGM ingredient derived from cottage cheese acid whey at 2, 5, and 10% (w/w). Each formulation underwent simulated digestion consisting of oral, gastric, and intestinal phases, and the micellar fraction was collected for both analysis and lipid assimilation in Caco-2 intestinal cells. The micellar fractions were diluted and applied to the cells for 4 h. A lipidomic approach was used to assess the lipid profiles of micellar fractions and intestinal cells. The micelles from digested jellies, cookies, and SF-cookies containing MFGM showed a distinct separation using partial least squares discriminant analysis (PLS-DA). Both correlation loadings and variable importance in projection (VIP) scores demonstrated a tendency of MFGM polar lipids (ceramides, glucosylceramides) for micelles from digested jelly, whereas micelles from digested cookies were associated with MFGM neutral lipids (free fatty acids, cholesterol, etc.). The effect of supplementation level on the micellar lipid profiles reinforced this pattern. The lipid profiles of intestinal cells after incubation with the micellar fractions differed considerably from the corresponding micellar lipid profiles. Specifically, the SF-cookie-treated cells were associated with a greater abundance of PUFA relative to jelly- and cookie-treated cells; however, increasing MFGM supplementation showed irregular patterns and rearrangement of cellular lipid profiles, suggesting the cells' role in regulating lipid metabolism in response to nutritional stimuli. The nature of lipid micellarization and assimilation in intestinal cells from MFGM-containing food formulations echoes the complexity of lipids inherent to the MFGM itself, suggesting the need for application-based MFGM supplementation.

Human Milk Composition and Nutritional Status of Omnivore Human Milk Donors Compared with Vegetarian/Vegan Lactating Mothers.

Women of childbearing age in Western societies are increasingly adopting vegetarian diets. These women are sometimes rejected as milk donors, but little about the composition of their milk is known. The present study aimed to compare the intake, nutritional status, and nutritional composition of human milk from omnivore human milk donors (Donors) and vegetarian/vegan lactating mothers (Veg). Milk, blood, and urine samples from 92 Donors and 20 Veg were used to determine their fatty acid profiles, as well as vitamins and minerals. In a representative sample of both groups, we also determined the lipid class profile as a distribution of neutral and polar lipids, the molecular species of triacylglycerols, and the relative composition of phospholipids in their milk. A dietary assessment was conducted with a five-day dietary record (while considering the intake of supplements). We highlight the following results, expressed as the mean (SE), for the Veg vs. Donors: (1) Their docosahexaenoic acid (DHA) intake was 0.11 (0.03) vs. 0.38 (0.03) g/day; the plasma DHA was 0.37 (0.07) vs. 0.83 (0.06)%; and the milk DHA was 0.15 (0.04) vs. 0.33 (0.02)%. (2) Their milk B12 levels were 545.69 (20.49) vs. 482.89 (4.11) pM; 85% of the Veg reported taking B12 supplements (mean dose: 312.1 mcg/day); and the Veg group showed no differences with Donors in terms of total daily intake or plasma B12. (3) Their milk phosphatidylcholine levels were 26.88 (0.67) vs. 30.55 (1.10)%. (4) Their milk iodine levels were 126.42 (13.37) vs. 159.22 (5.13) mcg/L. In conclusion, the Vegs' milk was shown to be different from the Donors' milk, mainly due to its low DHA content, which is concerning. However, raising awareness and ensuring proper supplementation could bridge this gap, as has already been achieved for cobalamin.

Lipidomic and Proteomic Profiling of the Milk Fat Globule Membrane from Different Industrial By-Products of the Butter and Butter Oil Manufacturing Process.

Recent studies have demonstrated the positive effects of regular intake of milk fat globule membranes (MFGMs) on neural and cognitive development, as well as immune and gastrointestinal health in infants and elders. Dairy products and by-products generated from the butter and butter oil manufacturing process are valuable sources of MFGM. Thus, in view of the growing need to reduce by-products and waste, it is crucial to foster research aimed at the valorization of dairy by-products rich in MFGM. For this purpose, all the by-products coming from butter and butter oil production (from raw milk to the related by-products) were used to study the MFGM isolated fractions, followed by their characterization through a combined lipidomic and proteomic approach. The patterns of polar lipids and proteins indicated that buttermilk (BM), butterserum (BS), and their mix (BM-BS blend) are the most suitable by-products to be employed as starting material for the isolation and purification of MFGMs, thus obtaining MFGM-enriched ingredients for the manufacture of products with high biological activity.

Milk fat globule membrane concentrate as a nutritional supplement prevents age-related cognitive decline in old rats: A lipidomic study of synaptosomes.

Aging is associated with a decline in cognitive abilities, mainly in memory and executive functioning. A similar but premature deterioration in cognitive capacities is the hallmark of mild cognitive impairment, Alzeimer's disease and dementia. The biochemical mechanisms that cause these neurodegenerative disorders are poorly understood. However, some evidence suggests that insufficient dietary intakes of some phospholipids could impact on brain function and increase the risk of future cognitive impairment and dementia. We evaluated the cognitive and biochemical effects of supplementation with a milk fat globule membrane (MFGM) concentrate in aged rats. We observed that, compared to control animals, MFGM supplemented rats showed enhanced spatial working memory, but both groups exhibited similar reference spatial learning and emotional memory abilities. No significant differences between BDNF levels in the hippocampus and frontal cortex of treated rats as compared to controls were found. The nootropic effects observed were accompanied by significant changes in the lipid composition of synaptic membranes. MFGM supplementation increased the levels of EPA and DHA acids as well as the plasmalogens content in the synaptosomes isolated from the hippocampus (Synapt-HP) and the frontal cortex (Synapt-FC). In addition enhanced levels of phosphatidyl serine (PS), particularly PS(18:1/18:1), and phosphatidyl inositol (PI) molecular species were observed in Synapt-HP and Synapt-FC of treated animals.Lipidomic analysis also revealed greater concentration of phosphatidyl ethanolamine (PE) molecular species containing very long-chain fatty acids and PE plasmenyls in Synapt-HP as well as an increase of the SM content in Synapt-FC from the MFGM group. Although further studies are needed to confirm the underlying mechanism (individual or synergistic), these results suggest that MFGM supplementation could be employed as a dietary implement to restore the proper cerebral concentration of some bioactive lipids and prevent or slow the progression of age-related cognitive impairment.

Randomized controlled trial of early arachidonic acid and docosahexaenoic acid enteral supplementation in very preterm infants.

Objective: To evaluate changes in blood long-chain polyunsaturated fatty acid (LCPUFA) and oxylipin concentrations in very preterm infants from birth to 36 weeks' postmenstrual age (WPA) after providing an emulsified arachidonic acid (ARA):docosahexaenoic acid (DHA) supplement at two different concentrations. Study design: This prospective, randomized trial assigned infants to receive a supplement (1) 80:40 group (80 mg/kg/day ARA and 40 mg/kg/day DHA, n = 9) or (2) 120:60 group (120 mg/kg/day ARA and 60 mg/kg/day DHA, n = 9). Infants received supplement daily from birth until 36 WPA. At baseline, 21 days of life and 36 WPA, the LCPUFAs were measured in plasma by gas chromatography/mass spectrophotometry. Additionally, LCPUFAs and oxylipins were analyzed in whole blood by ultra-high-performance liquid chromatography-tandem mass spectrometry. Furthermore, a sample of oral mucosa was obtained to analyze single-nucleotide polymorphism located in the FADS1 gene by PCR. Results: Gestational age was similar between groups (80:40 = 28+6 [27+3; 30+3] completed weeks+days ; 120:60 = 29+6 [27+3; 30+5] completed weeks+days , p = 0.83). At 36 WPA, the change in plasma ARA was significantly different between groups (80:40 group = 0.15 [-0.67; 0.69] %nmol, 120:60 = 1.68 [1.38; 3.16] %nmol, p = 0.031). In whole blood, the levels of ARA-derived oxylipins (5-, 8-, 9-, 11-, 15-HETE and 8,9-EET) and EPA-derived oxylipins (18-HEPE) significantly increase from baseline to 36 WPA in the 120:60 group than the 80:40 group. Conclusion: Supplementation at high doses (120:60 mg/kg/day) increased levels of ARA, and EPA- and ARA-derived oxylipins compared to low doses (80:40 mg/kg/day). Differences were detected in EPA metabolites without a significant increase in plasma DHA.

Impact of High-Pressure Processed Onion on Colonic Metabolism Using a Dynamic Gastrointestinal Digestion Simulator.

Onions are the main dietary source of flavonols that have been associated with important health-promoting properties. Onion treated by high-pressure processing (HPP-treated onion) was subjected to a dynamic gastrointestinal digestion and colon fermentation simulator (DGID-CF) to study the effect on the gut microbiota metabolism in the three colon regions (ascending-AC, transverse-TC, and descending-DC) by means of chronic feeding with 27 g/day for 14 days. HPP-treated onion presented a high content of the flavonols quercetin-3,4'-diglucoside and quercetin-4'-glucoside, and a large percentage of them reached the AC without change. TC and DC progressively increased the total phenolic metabolites 2.5 times respective to day 2, mainly 3-hydroxyphenylacetic, 4-hydroxyphenylacetic, 3-(4-hydroxyphenyl)-propionic, and 3,4-dihydroxyphenylpropionic acids. In addition, the chronic feeding increased the beneficial colon bacteria Bifidobacterium spp. and Lactobacillus spp. and the production of total SCFAs (acetic, propionic, and butyric acids) 9 times (AC), 2.2 times (TC), and 4.4 times (DC) respective to day 1. A multivariate analysis (principal component analysis, PCA) showed a clear separation between the three colon regions based on their phenolic composition (precursors and metabolites). These results showed that HPP-treated onion modulated the human gut microbiota's metabolism and the DGID-CF is a good system to study these changes.

Effect of gestational age (preterm or full term) on lipid composition of the milk fat globule and its membrane in human colostrum.

Human colostrum is the first milk secreted by the mother after birth and constitutes the ideal food for the newborn, because its chemical composition, rich in immunoglobulins, antimicrobial peptides, growth factors, bioactive lipids, and other important molecules, is perfectly adapted to the metabolic, digestive, and immunological immaturity of the newborn. An incomplete gestational period can affect the maturity of the mammary gland and its ability to secrete milk with the proper composition for the newborn's condition. Previous studies indicate that the mammary gland modulates the profiles of bioactive lipids present in the different phases of lactation from colostrum to mature milk. Given the key role played by the polar lipids (PL) (phospho- and sphingolipids) of the milk fat globule membrane (MFGM) in the immune system and cognitive development of the newborn, it is crucial to analyze whether the content and distribution of the PL are affected by gestation period. Therefore, this study aimed to determine the milk fat globule (MFG) and MFGM lipid compositions of human colostrum samples from 20 healthy preterm and full-term mothers. Lipid characterization using chromatographic techniques (gas chromatograph mass spectrometry and HPLC-evaporative light-scattering detection) revealed differences related to length of gestation in the profiles of lipid classes and fatty acid and triacylglyceride contents of colostrum. This comparative analysis leads to noteworthy outcomes about the changing roles of the PL, considering the preterm or full-term condition. We found a lack of correlation of some PL (such as phosphatidylcholine, phosphatidylinositol, and phosphatidylserine) with the delivery term; these could be denoted as structural category lipids. However, sphingomyelin and phosphatidyl-ethanolamine exhibited trends to decrease in full-term colostrum, indicating that in the final stage of pregnancy specific accretion of some PL occurs, which should be denoted as a nutritional redistribution.

Consumption of Goat Cheese Naturally Rich in Omega-3 and Conjugated Linoleic Acid Improves the Cardiovascular and Inflammatory Biomarkers of Overweight and Obese Subjects: A Randomized Controlled Trial.

This study examines the value of a goat cheese naturally enriched in polyunsaturated fatty acids (PUFA) (n-3 PUFA and conjugated linolenic acid (CLA)) as means of improving cardiovascular and inflammatory health. Sixty-eight overweight and obese subjects (BMI ≥ 27 and <40 kg/m2), with at least two risk factors for cardiovascular disease (CVD) in a lipid panel blood tests, participated in a randomized, placebo-controlled, double-blind, parallel designed study. The subjects consumed for 12 weeks: (1) 60 g/d control goat cheese and (2) 60 g/d goat cheese naturally enriched in n-3 PUFA and CLA. Diet and physical activity were assessed. Anthropometric and dual-energy X-ray absorptiometry (DXA) tests were performed. Blood samples were collected at the beginning and at the end of the study period. Changes in health status, lifestyle and dietary habits, and daily compliance were recorded. The consumption of a PUFA-enriched goat cheese significantly increased plasma high-density lipoprotein (HDL)-cholesterol, as well as in apolipoprotein B, and it significantly decreased high-sensitivity C-reactive protein concentrations compared to the control goat cheese (p < 0.05). The significant improvement of the plasma lipid profile and inflammatory status of people with risk for CVD due to the consumption of PUFA-enriched cheese suggests a potential role of this dairy product as an alternative to develop high nutritional value food in a balanced diet comprising regular exercise.

Appraisal of the suitability of two-stage extraction process by combining compressed fluid technologies of polar lipid fractions from chia seed.

Although triacylglycerols (TAG) are the major constituents of chia oil, it also contains minor lipid fractions that include phospholipids (PL) among other desirable components. Its amphiphilic character and excellent biocompatibility make PL appropriate for numerous applications with technological and nutritional significanceand potential health benefits. Given the difficulties entailed by the PL isolation, the efficiency for extracting such compounds using two environmental friendly techniques, pressurized liquid extraction (PLE) and supercritical fluid extraction (SFE) was evaluated. By using PLE with food-grade ethanol (EtOH), an oil recovery close to 100% was achieved in just 10 min. This oil extract was particularly rich in α-linolenic acid (ALA; 70%) as compared to the oil extracted by SFE (56%). In the case of SFE, the oil recovery was only 87% but increased to 99% when ethanol was added to CO2. However the use of co-solvent did not affect the fatty acid profile of the supercritical extracts or their TAG composition, where the high molecular weight TAG species were the predominant in all cases. With the exception of SFE without co-solvent, all methods applied were capable of extracting the PL fraction, although the content and distribution of the individual components present in this fraction differed markedly depending on the extraction conditions used. In this context, the use of a sequential extraction process, combining SFE and PLE was particularly interesting. The re-extraction by PLE of the chia cake, previously defatted by SFE, allowed to obtain an oil extract highly enriched in PLs, whose content exceeded 16% and with a higher PL species than the rest of the oil extracts.

The Influence of ß-Cyclodextrin on the Reduction of Cholesterol Content in Egg and Duck Liver Pâté.

The use of ß-cyclodextrin (ß-CD) to reduce cholesterol is increasing in food research due to its affinity for non-polar molecules such as cholesterol. The objective of this study was to study the ability of ß-CD to remove cholesterol in natural egg, powdered egg, and duck liver pâté and its effect on individual fatty acids. A concentration of 5% of ß-CD was found to be a suitable amount to remove 80.04 ± 4.96-82.12 ± 5.36% of cholesterol from yolk and powdered eggs and 80.21 ± 5.28% of cholesterol from duck liver pâté. ß-CD complexed to cholesterol was precipitated and removed by centrifugation. Individual fatty acid compositions did not differ (p < 0.05) between the controls and the products treated with 5% ß-CD.

Alterations in the Fatty Acid Composition in Infant Formulas and ω3-PUFA Enriched UHT Milk during Storage.

ω3-polyunsaturated fatty acids (PUFA) are known to have several beneficial effects, such as preventing the occurrence of cardiovascular events in adults and improving the development of the central nervous system during fetal life and childhood. Dairy products enriched in PUFA are now available on the market and can help consumers to meet the ω3-PUFA daily intake recommendations. Although PUFA are prone to oxidation, little information exists regarding the alterations they undergo when products containing PUFA are stored. In this study, five infant formulae (IF) and five ω3-PUFA enriched Ultra High Temperature (UHT)-milk products were examined during storage at room temperature in terms of fatty acid (FA) composition and trans fatty acid (tFA) content until the end of shelf life by chromatography techniques. The IF included two follow-on formulae, two first-age formulae and a special medical purpose formula with different fat contents (from 2.6% to 27.7%). In the ω3-PUFA enriched UHT-milk products the milk fat was replaced by eicosapentanoic fatty acid (EPA) and docosahexaenoic acid (DHA) rich oils. The fatty acid content of all samples remained stable whereas some variations were observed for the tFA isomer content (p < 0.05) in the UHT-milk samples.

Production of omega 3-rich oils from underutilized chia seeds. Comparison between supercritical fluid and pressurized liquid extraction methods.

Chia seeds constitute a promising source of α-linolenic acid (ALA). In the present work, an underutilized and cheaper set of chia seeds, which were discarded after the harvest according to quality criteria - named in this work as low oil content seeds (LOCS) - have been evaluated as a potential source for obtaining PUFA-enriched oils against the commonly studied high-quality chia seeds denoted as high oil content seeds (HOCS) in this study. Two efficient and environmental friendly techniques, supercritical fluid extraction (SFE) and pressurized liquid extraction (PLE), were evaluated to optimize the extraction process of chia oil. At 60 °C, by using pressurized food-grade ethanol, recoveries close to 100% were achieved from both sets of seeds in a short extraction time (10 min). By using SFE, the greatest oil extraction yield (>95%) was attained at the highest pressure and temperature conditions (45 MPa and 60 °C) after 240 min. At the early stage of SFE extraction, both LOCS and HOCS exhibited a similar kinetic behavior, reaching oil extraction rates of 0.59 g oil/min and 0.64 g oil/min, respectively. No differences were found between the fatty acid profile of the oils extracted from LOCS and HOCS both by PLE and SFE. ALA and linoleic acid (LA) concentrations ranged between 65-68% and 17-23% respectively, and a predominance of high molecular weight triglycerides (≥ CN50), was found in all extracted oils. In conclusion, LOCS might constitute a new suitable raw material for the production of ALA-enriched oils. Concerning the extraction methods assayed, the oil was almost entirely recovered by both PLE and SFE at the used conditions.

Effect of Beta Cyclodextrin on the Reduction of Cholesterol in Ewe's Milk Manchego Cheese.

Beta-cyclodextrin (ß-CD) is a cyclic oligosaccharide consisting of seven glucose units and is produced from starch using cyclodextrin glycotransferase enzymes to break the polysaccharide chain and forming a cyclic polysaccharide molecule. The use of ß-CD in food research for reduction of cholesterol is increasing due to its affinity for non-polar molecules such as cholesterol. The aim of this study was to evaluate the feasibility of using ß-CD in cholesterol removal from pasteurized ewe's milk Manchego cheese and evaluate the effect on the main components of the milk, lipids, and flavor characteristics. Approximately 97.6% cholesterol reduction was observed in the cheese that was treated using ß-CD. Physicochemical properties (fat, moisture and protein) were not changed by the ß-CD treatment, except the soluble nitrogen and non-protein nitrogen that showed slight differences after the treatment. The amount of the different components of the lipid fraction (fatty acids, triglycerides and phospholipids) were similar in cheeses treated and not treated with ß-CD. Flavor compound and short chain free fatty acids were not mostly significantly influenced by the effect of the ß-CD. ß-CD molecules are edible and nontoxic and as a result they can be used safely for cholesterol removal processing in cheese manufacturing. Therefore, the present study suggests that ß-CD treatment is an effective process for cholesterol removal from Manchego cheese while preserving its properties.

Reduced-intensity conditioning allogeneic transplantation from unrelated donors: evaluation of mycophenolate mofetil plus cyclosporin A as graft-versus-host disease prophylaxis.

In the current study, we have analyzed the efficacy of cyclosporine A (CSA) plus mycophenolate mofetil (MMF) as graft-versus-host disease (GVHD) prophylaxis in the fludarabine plus melphalan or busulfan reduced intensity regimen (RIC) setting in a series of 44 patients receiving allogeneic transplantation from an unrelated donor. Only 23% were in the first complete remission at the time of transplant. Cumulative incidence of grades II-IV and III-IV acute GVHD (aGVHD) was 53% and 23%, respectively. Fifty-six percent had equal to or greater than grade 2 gut involvement. Cumulative incidence of overall and extensive chronic GVHD (cGVHD) was 93% and 63%, respectively. Ninety-two percent of patients who were evaluable +100 days after transplant were in complete remission. Relapse rate was 25% at 2 years. Event free (EFS) and overall survival (OS) at 2 years were 52%. Pharmacokinetic assays of mycophenolic acid (MPA) showed a therapeutic area under the curve (AUC) at the dosage of 3 g daily, although a large inter- and intraindividual variations of MPA plasma levels were found. In conclusion, the combination of CSA plus MMF in the fludarabine plus melphalan or busulfan RIC setting is feasible. Regarding GVHD, this combination allowed to control aGVHD but lead to a high incidence of cGVHD, so that newer strategies are required, especially in trying to decrease gastrointestinal involvement.