Carlos Muñoz

Effects of a nutritional supplement on the immune response and cytokine production in free-living Chilean elderly.

BACKGROUND Immune response is impaired in the elderly. Our aim was to study the effects of a special nutritional formula on the immune response and response to influenza and pneumococcal vaccination in elderly subjects. METHODS Sixty healthy subjects aged > or = 70 years, with a Mini Mental score > or = 22 were studied. Half of the subjects received a special nutritional formula (in addition to the regular diet) providing, among other nutrients, 480 kcal, 31 g proteins, 120 IU vitamin E, 3.8 microg vitamin B12, 400 microg folic acid, 10(9) cfu Lactobacillus paracasei (NCC 2461), and 6 g of fructo-oligosaccharides. At 4 months of follow-up, subjects were vaccinated against influenza and pneumococcus. Lymphokine production by mononuclear cells (PBMC), lymphocyte subpopulations, and natural killer cell (NK) activity were measured at baseline and 4 months of follow-up (before vaccination). Antibodies against influenza and pneumococcal antigens and flu-stimulated production of interferon gamma and interleukin-2 by PBMC were measured at 4 and 6 months. Skin response to 7 recall antigens and body composition were assessed at baseline and at 4 and 12 months. All infections occurring during the study period were recorded. RESULTS NK activity increased in supplemented subjects and decreased in nonsupplemented individuals. Interleukin-2 production by PBMC and the proportion of T cells with NK activity decreased in controls and did not change in supplemented subjects. Supplemented subjects reported less infections than nonsupplemented individuals (in 13% and 22% of scheduled visits, respectively; p = .02). CONCLUSIONS This nutritional supplement increased innate immunity and protection against infections in elderly people.

Effects of prebiotics on the immune response to vaccination in the elderly

BACKGROUND Prebiotics stimulate the growth of bifidogenic bacteria in the gut. The aim of this work was to assess the effects of a prebiotic mixture on the immune response in healthy elderly people. METHODS Healthy free-living elderly people (age, > or = 70 years), receiving a nutritional supplement that provided 1.6 MJ, 15 g of protein, and 50% of vitamin daily reference values per day, were randomly assigned to receive a prebiotic mixture (6 g/d of a 70% raftilose and 30% raftiline mixture) or placebo (6 g of maltodextrin powder) for 28 weeks. At week 2 of the study, all subjects were vaccinated with influenza and pneumococcal vaccines. At weeks 0, 2, and 8 of the study, serum total proteins, albumin, immunoglobulins, saliva secretory immunoglobulin A (IgA), and serum titers of influenza A and B and pneumococcal antibodies were measured. At week 8, cultured peripheral monocyte cell secretion of interleukin-4, interferon-gamma, and lymphocyte proliferation, stimulated with phytohemagglutinin and influenza antigen, were measured. RESULTS Sixty-six subjects were considered eligible for the study, and 43 (20 receiving prebiotics and 23 receiving placebo) were considered for final analyses on a per protocol basis. No changes in serum proteins, albumin, immunoglobulins, and secretory IgA were observed. Antibodies against influenza B and pneumococcus increased significantly from weeks 0 to 8, with no significant differences between groups. Antibodies against influenza A did not increase. No effects of prebiotics on interleukin-4 and interferon-gamma secretion by cultured monocytes were observed. CONCLUSIONS No immunological effects of prebiotics were observed in this study.

Nutritional Support in Alcoholic Cirrhotic Patients Improves Host Defenses

BACKGROUND Malnutrition is usual in patients with alcoholic liver disease and is associated with a poor outcome. Nutritional support decreases nutrition-associated complications. AIM To demonstrate that nutritional support in ambulatory alcoholic cirrhotic patients improves host defenses. METHODS Thirty-one male outpatients with alcoholic cirrhosis CHILD-PUGH B or C were included. Twenty-five subjects completed six months consuming daily a nutritional supplement (Ensure, 1000 Kcal and 35 g protein), in addition to their regular diet. At entrance and every three months, a clinical assessment, nutritional evaluation and indirect calorimetry were performed. Liver function tests and LPS-induced monocyte production of cytokines, salivary secretory IgA, lactulose/mannitol ratio and breath hydrogen tests were also measured in these intervals. Delayed cutaneous hypersensitivity and IgG and IgM antibody response to endotoxin were assessed at entrance and at the end of the study. RESULTS Patients drank 85% of the provided supplement as an average. REE, total body fat and serum albumin increased, basal breath hydrogen decreased and cellular immunity improved significantly during the follow up period (p< or =0.03). All the other parameters remained unchanged throughout the study. Six patients (16.2%) died during the study, five due to upper gastrointestinal bleeding. CONCLUSION Nutritional support in alcoholic cirrhotic patients improves nutritional status and cell mediated immunity.

Iron, copper and immunocompetence.

Microminerals including copper and iron are essential to immunity and health in human beings. The development of powerful tools in analytical cell biology and molecular genetics has facilitated efforts to identify specific cellular and molecular functions of trace elements in the maturation, activation and functions of host defence mechanisms. Selected recent reports about the role of copper and iron nutrition on immune functions are critically analysed here. Effects of trace element supplementation on infectious morbidity are also reviewed. While micromineral deficiencies, in general, may have widespread effects on nearly all components of immune response, these effects can be reversed by supplementation. However, the conflicting effects of iron deficiency and iron supplementation in vitro on the defensive systems reveals the urgent need for further additional information on the in vivo situation. In the elderly, vaccination against respiratory infections is likely to protect only 30-70% of the population. However, it may be possible to modulate immune function and ultimately reduce the severity of infections through micronutrient supplementation. Thus, microminerals contribute to the maintenance of the balance between immunity and health in humans.

Impaired interleukin-1 and tumor necrosis factor production in protein-calorie malnutrition

Abstract The capacity of malnourished infants to produce interleukin-1 beta (IL-1β) and tumor necrosis factor alpha (TNF-α) was evaluated before and after nutritional rehabilitation. Ten marasmic patients without infectious diseases 2 to 8 months of age with −2.48±0.54 weight for age Z score (mean ± SD) were studied on admission, and after 4 month of nutritional therapy in a Closed Nutritional Recovery Center. Cytokines were induced by in vitro stimulation of blood mononuclear cells with lipopolysaccharide. IL-1β and TNF-α were determined in supernatants and cell lysates using an specific immunoassay. Low levels of released and cell-associated IL-1β were found on admission; mean±SEM values were 2.0±0.5 and 1.3±0.5 ng/ml, respectively. After nutritional rehabilitation, a significant increase of released and cell-associated IL-1β were observed: 5.4±0.8 and 3.7±0.3 ng/ml, (mean±SEM) respectively, p

Local and systemic liberation of proinflammatory cytokines in ulcerative colitis

Determination of plasma and tissue cytokinelevels in inflammatory bowel disease have frequentlyresulted in conflicting data. In the present study wedetermined in patients with ulcerative colitis (UC), the levels of the proinflammatory cytokinesinterleukin (IL)-1β, IL-6, interferon(IFN)-γ, and tumor-necrosis factor (TNF)-αliberated by peripheral blood mononuclear cells (PBMC)and lamina propria mononuclear cells (LPMC) after 48-hrculture with pokeweed mitogen (PWM). IL-1β, IL-6,IFN-γ and TNF-α in the supernatant weredetected by ELISA. Results show low basal levels ofIL-1β secretion by PBMC and LPMC, and a considerableincrease after mitogen stimulation. Basal IL-6production by PBMC was higher in UC patients than incontrols [2029 pg/ml, CI9 (–165 to4223) vs 572 pg/ml (–383 to 1527) respectively, P = 0.05] and also afterPWM activation [14,995 pg/ml (7759 -22230) vs 6598 pg/ml(3240-9956), respectively, P = 0.05]. In LPMC, nodifferences in IL-6 secretion were observed. TNF-α in activated PBMC of patients with UC was notsignificantly increased in relation to control (P =0.09). No constitutive secretion of IFN-γ wasobserved in mononuclear cells. IFN-γ levelssecreted by activated LPMC were lower in patients withUC than in controls [1571 pg/ml (–108 to 3251) vs7953 pg/ml (3851-12,055), respectively, P = 0.03]. Theseresults suggest that IL-6, IL-1β, and TNF-α participate as mediators in the inflammatoryphenomena observed in UC. Further studies are necessaryto evaluate the role of IFN-γ in thiscondition.

Construction of High Density Sweet Cherry (Prunus avium L.) Linkage Maps Using Microsatellite Markers and SNPs Detected by Genotyping-by-Sequencing (GBS)

Linkage maps are valuable tools in genetic and genomic studies. For sweet cherry, linkage maps have been constructed using mainly microsatellite markers (SSRs) and, recently, using single nucleotide polymorphism markers (SNPs) from a cherry 6K SNP array. Genotyping-by-sequencing (GBS), a new methodology based on high-throughput sequencing, holds great promise for identification of high number of SNPs and construction of high density linkage maps. In this study, GBS was used to identify SNPs from an intra-specific sweet cherry cross. A total of 8,476 high quality SNPs were selected for mapping. The physical position for each SNP was determined using the peach genome, Peach v1.0, as reference, and a homogeneous distribution of markers along the eight peach scaffolds was obtained. On average, 65.6% of the SNPs were present in genic regions and 49.8% were located in exonic regions. In addition to the SNPs, a group of SSRs was also used for construction of linkage maps. Parental and consensus high density maps were constructed by genotyping 166 siblings from a ‘Rainier’ x ‘Rivedel’ (Ra x Ri) cross. Using Ra x Ri population, 462, 489 and 985 markers were mapped into eight linkage groups in ‘Rainier’, ‘Rivedel’ and the Ra x Ri map, respectively, with 80% of mapped SNPs located in genic regions. Obtained maps spanned 549.5, 582.6 and 731.3 cM for ‘Rainier’, ‘Rivedel’ and consensus maps, respectively, with an average distance of 1.2 cM between adjacent markers for both ‘Rainier’ and ‘Rivedel’ maps and of 0.7 cM for Ra x Ri map. High synteny and co-linearity was observed between obtained maps and with Peach v1.0. These new high density linkage maps provide valuable information on the sweet cherry genome, and serve as the basis for identification of QTLs and genes relevant for the breeding of the species.

Pravastatin immunomodulates IL-6 and C-reactive protein, but not IL-1 and TNF-α, in cardio-pulmonary bypass

BACKGROUND While statins are increasingly used in cardiopulmonary bypass (CPB), the anti-inflammatory effects of individual statins, within the context of various treatment regimes, need further examination. The present study evaluates the anti-inflammatory effectiveness of the short-term, preoperative and intensive postoperative use of pravastatin in CPB. METHOD Forty three patients undergoing CPB were enrolled in a randomized, prospective clinical study. One group (n = 21), received pravastatin, the other (n = 22) did not. Patients in the pravastatin group received one dose of 40 mg per day for nine days, starting 48 hours before CPB, with an additional dose of 40 mg one hour after surgery. Plasma levels of selected inflammatory mediators were measured at baseline and tracked systematically. RESULTS Pravastatin reduced postoperative interleukin-6 (IL-6) levels significantly at 24 and 48 hours, and at seven days. Mean +/- SD values, for treated versus untreated patients were: at 24 hours, 159.5 +/- 58.5 versus 251.2 +/- 53.0 pg/mL (p < 0.001); at 48 hours, 81.9 +/- 31.5 versus 194.2 +/- 56.3 pg/mL (p < 0.001); and at seven days, 16.4 +/- 7.2 versus 30.8 +/- 12.6 (p < 0.001). C-reactive protein (CRP) decreased significantly on the seventh postoperative day, when plasma levels were 3.6 +/- 1.1 in the treated patients versus 8.2 +/- 2.1 mg/dL in the controls (p < 0.001). No changes in plasma IL-1 and TNF-alpha were found during entire study. CONCLUSIONS Pravastatin induced a precocious modulation of IL-6 expression and a later reduction of plasma CRP levels. Pravastatin;s effects on the expression of these pivotal inflammatory mediators strongly support its well-timed use in CPB.

Rotating disk sorbent extraction for pre-concentration of chromogenic organic compounds and direct determination by solid phase spectrophotometry

A novel and very simple microextraction approach for pre-concentration and direct solid phase spectrophotometric measurement has been developed for the determination of chromogenic analytes. The model analyte to assess this approach was the chromophore malachite green (MG). The analyte was extracted from water samples onto a small rotating disk made of Teflon containing a sorbent phase of polydimethylsiloxane (PDMS) on one of its surfaces. We refer to the extraction procedure as rotating disk sorptive extraction (RDSE). After extraction, the sorbent phase with the concentrated analyte was separated from the Teflon disk and used directly for MG determination by solid phase spectrophotometry at 624 nm, without the necessity of a desorption step. Chemical and extraction variables such as concentration of sodium sulfate, pH, disk rotational velocity, extraction time, and temperature were studied in order to establish the best conditions for extraction. Under optimum conditions, the extraction of MG was carried out in 18 min and 90 min, for sample volumes of 100mL or 1000 mL, respectively. The detection limit, based on three times the standard deviation of the blank phase (3σ(b)), was 1.4 μg L⁻¹ and the repeatability, expressed as relative standard deviation (RSD), for 20 μg L⁻¹ MG was 8.1%. This study also applied the method to real samples, obtaining quantitative recovery (mean recovery of 99.3%). The PDMS phases could be reused after desorbing the MG into methanol for 3h. Replacement of the PDMS film onto the disk is very easy and low cost.

Phagocytosis and immunoglobulin levels in hypocupremic infants

Abstract Nineteen hypocupremic marasmic infants 5–14 months of age with a normal wt/1gth ratio, were immunologically evaluated during their rehabilitation. The infants were studied 2–3 months after admission to a Closed Nutritional Recovery Center and subsequently after copper supplementation. Two groups were defined: a severely copper deficient group (A), with abnormal copper and ceruloplasmin levels; and a marginal copper deficient group (B), with abnormal plasma copper level and normal serum ceruloplasmin. Immunoglobulin levels were measured by radial immunodiffusion technique and phagocytosis to S. aureus by using autologous and normal homogogous plasma. After copper supplementation plasma copper increased to normal values in both groups of infants. Ceruloplasmin exceeded normal ranges in group A whereas no changes were detected in group B. Serum immunoglobulins were normal and secretory IgA remained low in most of the infants. Phagocytic indices increased in most infants with both types of plasma after copper supplementation in groups A and B (p