Efficacy of Bifidobacterium breve CECT7263 for infantile colic treatment: an open-label, parallel, randomised, controlled trial.

Infantile colic is a prevalent condition characterised by excessive crying with no effective treatment available. We aimed to evaluate the efficacy of Bifidobacterium breve CECT7263 and a combination of this and Lactobacillus fermentum CECT5716 versus simethicone in reducing the daily time spent crying in colicky infants. A multicentre randomised, open-label, parallel, controlled trial of 28 days was performed in 150 infants who were diagnosed with colic according to the Rome III criteria and who randomly received simethicone (80 mg/day; Simethicone group), B. breve CECT7263 (2×108 cfu/day, Bb group), or a combination of L. fermentum CECT5716 and B. breve CECT7263 (1×108 cfu/day per strain, Bb+Lf group). The main outcomes were minutes of crying per day and the percentage of reduction in daily crying from baseline. Data were analysed per intention to treat. All treatments significantly decreased the daily crying time at the end of the intervention (P-time <0.001). However, the infants in the Bb group had significantly decreased crying time from the first week of the study (P<0.05), whereas the Bb+Lf group and the simethicone group had significantly decreased crying time from the second week (P<0.05). The percentage of reduction in the minutes of crying from baseline in the Bb group was significantly higher than that in the Simethicone group every week of the intervention (-40.3 vs -27.6% at 1-week; -59.2 vs -43.2% at 2-weeks; -64.5 vs -53.5% at 3-week and -68.5 vs -59.5% at 4-weeks, P<0.05). Additionally, in the Bb group, infants had better night sleep, and parents reported a more positive mood at the end of the intervention. All the products used in the study were safe and well tolerated. In conclusion, the breastmilk-isolated probiotic strain B. breve CECT7263 is a safe and effective treatment for infantile colic, presenting an earlier and more robust effect than the reference prescribed drug, simethicone.

Effects of Lactobacillus fermentum CECT5716 Lc40 on infant growth and health: a randomised clinical trial in nursing women.

The breast milk microbiota has been described as a source of bacteria for infant gut colonisation. We studied the effect of Lactobacillus fermentum CECT5716 (Lc40) on growth and infection incidence of the infants, when the probiotic is administrated to the mothers. Moreover, whether such effects might depend on the interaction between the mother or infant microbiota and the probiotic administration. A total of 291 mother-infant pairs were studied for 16 weeks in a randomised double-blinded placebo-controlled multicentre trial. The Lc40 group (n=139) received 1 capsule/day containing 3×109 cfu Lc40; the control group (n=152) received 1 placebo (maltodextrin) capsule/day. A positive and significant correlation of the Staphylococcus load between breast milk and infant faeces was only observed in control group. Additionally, the weight z-score of the infants whose mothers had higher values of Lactobacillus in their breast milk were significantly higher for the Lc40 group. We observed a significant lower incidence of conjunctivitis in the infants whose mothers received Lc40. A higher load of Staphylococcus in infant faeces significantly increased the risk of respiratory infections. Such incidence, under an absent or low Staphylococcus load in the faeces, was significantly 36 times higher in the infants in the control group than in the infants in the Lc40 group. However, the protective effect of Lc40 was gradually reduced as the Staphylococcus load of the milk increased. The administration of Lc40 to nursing women might influence infant growth and health but it seems to depend on its interactions with mother or infant microbiota. Registered in the US Library of Medicine (www.clinicaltrials.gov): NCT02203877.

Long-term safety of early consumption of Lactobacillus fermentum CECT5716: A 3-year follow-up of a randomized controlled trial.

Lactobacillus fermentum CECT5716 is a probiotic strain originally isolated from human breast milk. Previous clinical studies in infants showed that the early administration of a milk formula containing this probiotic strain was safe and may be useful for the prevention of community-acquired infections. This is a 3-year follow-up study aimed at evaluating the long-term effects produced by the early consumption of an infant formula supplemented with L. fermentum CECT5716 (experimental group, EG) compared with a control formula without the probiotic (control group, CG). The infants included in this follow-up study had previously completed a 5-month randomized double-blind controlled trial (from 1 to 6 months of age), where the safety and tolerance of the probiotic formula was evaluated. The main outcome of the follow-up study was the growth of the children. The secondary outcomes included the incidence of infectious and non-infectious diseases, parameters related with intestinal function and faecal microbiota. At 3 years, the mean values of weight, length and head circumference were similar in children of the EG compared with those of the CG. No differences were observed in the incidence of infectious and non-infectious diseases or disorders related with intestinal function. The pattern of faecal microbiota was also similar between both groups. In conclusion, this 3-year study shows that the early administration of the probiotic of L. fermentum CECT5716 in an infant formula is safe and it does not produce measurable differences in children compared with a control formula.

Daily consumption of milk enriched with fish oil, oleic acid, minerals and vitamins reduces cell adhesion molecules in healthy children.

BACKGROUND AND AIMS: Several studies have suggested that polyunsaturated fatty acids, vitamins and minerals have beneficial effects on lipid profile and systemic inflammation in adults. METHODS AND RESULTS: We examined the effects of a daily intake of milk enriched with longchain polyunsaturated fatty acids, oleic acid, carbohydrates, vitamins, minerals and low in saturated fatty acids (SFAs) for 5 months, on several cardiovascular (CVD) risk biomarkers in healthy children aged 8-14 years. In a randomized double-blind placebo-controlled trial, a total of 107 children of both genders were assigned to two study groups: 1) a supplemented group (SG, n=53) who consumed 0.6 L/day of an enriched dairy product, and 2) a control group (CG, n=54) who consumed 0.6 L/day of standard whole milk. Both groups consumed the dairy drinks for 5 months, in addition to their usual diet. Serum levels of adhesion molecules as indices of vascular endothelial cell activation were assessed in both groups at 0 and 5 months as well as white blood cell counts, lipid profile, serum proteins, total serum calcium, 25-OH vitamin D, glucose, insulin and adiponectin. In the enriched dairy drink supplemented group, adhesion molecules E-selectin and ICAM-1 as well as lymphocyte levels decreased while plasma docosahexaenoic acid (DHA) and serum calcium concentrations increased. In the control group, serum total protein, transferrin, total cholesterol, HDL-cholesterol and adiponectin concentrations decreased. CONCLUSION: The consumption of a milk enriched with fish oil, oleic acid, minerals and vitamins reduced indices of endothelial cell activation in the studied group of healthy children.

One-month administration of hydroxytyrosol, a phenolic antioxidant present in olive oil, to hyperlipemic rabbits improves blood lipid profile, antioxidant status and reduces atherosclerosis development.

The present study analysed the effects of hydroxytyrosol (HT) on blood lipids, antioxidant status and the progression of aortic lesions in hyperlipemic rabbits. Sixty-four rabbits were distributed into eight groups of animals (n = 8). Animal groups C, A and H were fed for 1-month with a control diet containing sunflower oil (C), an atherogenic diet (A) high in saturated fat and cholesterol or the A diet together with HT, respectively. The other five groups were fed for 2-months with diets C or A (groups CC or AA, respectively), or for 1-month with the A-diet followed by a further month with diet C, extra virgin olive oil diet (O) or diet C with HT (groups AC, AO and AH, respectively). Four milligram of HT/kg body weight were used in the study. Fifty and 42% decrease in total cholesterol and triacylglycerols, respectively, and a 2.3-fold increase in HDL-cholesterol were observed in the AH group but not in the H group. The HT-supplemented groups improved their antioxidant status and reduced the size of atherosclerotic lesions measured as intimal layer areas of the aortic arch when compared with control animals. We conclude that HT supplementation may have cardioprotective effects in vivo.

[Cardiovascular effects of omega-3-fatty acids and alternatives to increase their intake]. / Efectos cardiovasculares de los ácidos grasos omega-3 y alternativas para incrementar su ingesta.

Cardiovascular diseases are the main mortality cause in Europe, the USA and a great extent of Asia. There are several risk factors associated with cardiovascular diseases, such as increased total cholesterol, homocysteine and triglycerides, hypertension, diabetes, and reduced levels of HDL-cholesterol. Many of these risk factors are diet influenced. In spite of the great amount of foods enriched with n-3 fatty acids available at the market, the knowledge about the effects produced by regular intake of these foods still is a challenge in the majority of cases. It appears that intake of foods enriches with n-3 polyunsatured fatty acids is an option that may be effective in reducing risk factors for diseases, by substituting supplements without modifying consumer's alimentary habits. Also shown are the outcomes from a nutritional study undergone with a functional milk-bases food that contains n-3 fatty acids, oleic acid and vitamins.

Efectos cardiovasculares de los ácidos grasos omega-3 y alternativas para incrementar su ingesta / Cardiovascular effects of omega-3-fatty acids and alternatives to increase their intake

Las enfermedades cardiovasculares son la principal causa de mortalidad en Europa, Estados Unidos y gran parte de Asia. Existen varios factores de riesgo asociados a las enfermedades cardiovasculares, entre ellos están el colesterol total, la homocisteína y los triglicéridos elevados, la hipertensión, la diabetes y niveles reducidos de colesterol HDL. Muchos de estos factores de riesgo son influenciables por la dieta. A pesar de la gran cantidad de alimentos enriquecidos en ácidos grasos n-3 disponibles en el mercado, el conocimiento de los efectos originados por el consumo regular de estos alimentos supone aún un reto en la mayoría de los casos. La ingesta de alimentos enriquecidos en ácidos grasos poliinsaturados n-3 parece ser una opción que puede ser eficaz en la reducción de factores de riesgo de enfermedades, sustituyendo a los suplementos sin originar cambios en los hábitos alimentarios del consumidor. También se muestran los resultados procedentes de un estudio nutricional que hemos llevado a cabo con un alimento funcional de base láctea que contiene ácidos grasos n-3, ácido oleico y vitaminas (AU)

Cardiovascular diseases are the main mortality cause in Europe, the USA and a great extent of Asia. There are several risk factors associated with cardiovascular diseases, such as increased total cholesterol, homocysteine and triglycerides, hypertension, diabetes, and reduced levels of HDL-cholesterol. Many of these risk factors are diet influenced. In spite of the great amount of foods enriched with n-3 fatty acids available at the market, the knowledge about the effects produced by regular intake of these foods still is a challenge in the majority of cases. It appears that intake of foods enriches with n-3 polyunsatured fatty acids is an option that may be effective in reducing risk factors for diseases, by substituting supplements without modifying consumer's alimentary habits. Also shown are the outcomes from a nutritional study undergone with a functional milk-bases food that contains n-3 fatty acids, oleic acid and vitamins (AU)

The teaching of human Anatomy in Spain at the beginning of the twenty-first century

This work, which merely aims to reflect theauthors’ opinions, begins by pointing out a seriesof problems involved in the teaching of HumanAnatomy in Spanish Universities over the pastquarter century. Emphasis is placed on both thecauses that can be attributed to anatomists themselvesand on those deriving from Universitylegislation, especially after the University ReformLaw. We also refer to the small body of literaturepublished by Spanish anatomists with respectto the teaching of Anatomy, cadaver dissection orthe use of new technologies in such teaching. Wethen offer an overview of the current circumstancesand future possibilities not only of teachingand methodology but also as regards instructorsand departments. We offer a reference tothe Spanish Anatomical Society and we end byproposing a series of guidelines for the immediatefuture with a view to optimising the teachingof Human Anatomy and Embryology andthe professionalisation of future anatomists

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Magnetic resonance microscopy versus light microscopy in human embryology teaching.

A study was carried out on the application of magnetic resonance microscopy (MRM) in teaching prenatal human development. Human embryos measuring 8 mm, 15 mm, 18.5 mm, and 22 mm were fixed in a 4% paraformaldehyde solution and sections obtained with magnetic resonance imaging (MRI) were compared to those prepared for light microscopy (LM), using the same embryos. The MRM and LM slices were of a similar quality. In the MRM sections, embryonic organs and systems were clearly visible, particularly the peripheral and central nervous systems, and the cardiovascular and digestive systems. The digitalization and clarity of the MRM images make them an ideal teaching aid that is suitable for students during the first years of a health-science degree, particularly medicine. As well as providing students with their first experience of MRM, these images allow students to access, at any time, all embryos used, to assess changes in the positions of different organs throughout their stages of development, and to acquire spatial vision, an absolute requirement in the study of human anatomy. We recommend that this technique be incorporated into the wealth of standard embryonic teaching methods already in use.

n-3 Fatty acids plus oleic acid and vitamin supplemented milk consumption reduces total and LDL cholesterol, homocysteine and levels of endothelial adhesion molecules in healthy humans.

BACKGROUND AND AIMS: Numerous studies suggest n -3 polyunsaturated fatty acids (n -3 PUFA) and oleic acid intake have beneficial effects on health including risk reduction of coronary heart disease. The purpose of this study was to evaluate the effect of a commercially available skimmed milk supplemented with n -3 PUFA, oleic acid, and vitamins E, B(6), and folic acid (Puleva Omega3) on risk factors for cardiovascular disease. (CVD). METHODS: Thirty volunteers were given 500 ml/day of semi-skimmed milk for 4 weeks and then 500 ml/day of the n -3 enriched milk for 8 further weeks. Plasma and LDL lipoproteins were obtained from volunteers at the beginning of the study (T(pre)), and at 4, 8 and 12 weeks. RESULTS: The consumption of n -3 enriched milk produced a significant decrease in plasma concentration of total and LDL cholesterol accompanied by a reduction in plasma levels of homocysteine. Plasma and LDL oxidability and vitamin E concentration remained unchanged throughout the study. A significant reduction in plasma levels of vascular cell adhesion molecule 1, and an increase in plasma concentration of folic acid were also observed. CONCLUSION: Daily intake of n -3 PUFA and oleic acid supplemented skimmed milk plus folic acid and B-type vitamins has favourable effects on risk factors for CVD.

The human embryo development through MMR.

This study evaluates the use of the Microscopic Magnetic Resonance (MMR) in the human prenatal development. Human embryos (8mm, 15mm, 18mm and 22mm in length) fixed in 4% paraformaldehyde were used. Results were compared with light microscopy (LM) images. The internal configuration of the embryos can be clearly observed as well as many organs such as liver, lungs, heart, including their spatial relationships. In general MMR sections are less clear and show minor details than those by LM. Neverthless, many advantages are provided by using this technique. For example it is possible: a) to make three-dimensional (3-D) surface and internal full or partial reconstructions; b) to evaluate the presence of developmental anomalies; c) to evaluate the tissular preservation degree of the specimens; and d) to apply morphometric techniques to unfixed specimens. In our opinion the advantages derived by using MMR are many and overcome the disadvantages. This study demonstrates that MMR can be incorporated into ordinary laboratory techniques in human development studies, being also an initial election technique opposite to others more aggressive.

Immunocytochemical study on the triple origin of the sphincter iris in the chick embryo.

The ontogenic development of the sphincter iris has been studied by immunocytochemistry and standard staining on chick embryos from stage 25 HH to the time of hatching. We have used the monoclonal antibody 13F4, a highly specific marker of muscular cells. We have observed three different regions in the iris. In the pupillary region, immunoreactive cells are in continuous contact with the inner epithelium of the pupillary margin. In the intermediate region, the outer epithelium forms buds of pigmented cells that emigrate toward the stroma. In this epithelium cells that are totally or partially unpigmented exist, and they are 13F4 positive. In the sphincter we have observed 13F4 positive cells with melanin granules. In the ciliary region, the immunoreactivity appears in dispersed mesenchymal cells. The present findings are consistent with a triple origin of the sphincter iris in the chick embryo. This muscle is derived from the inner epithelium of the pupillary margin, the intermediate region of the outer epithelium, and from the mesenchymal cells. The cells of the inner epithelium of the pupillary margin are differentiated into smooth muscle cells, and the remaining cells form striated muscle cells.

Anomalias cardíacas producidas por disminución de flujo sanguíneo en embriones de pollo / Cardiac produced by disminution of the blood flow in chicken embryos

En un intento por conocer experimentalmente la importancia de los factores hemodinámicos en la morfogénesis cardíaca, diseñamos una investigación para determinar las consencuencias que ejerce la disminución del flujo sanguíneo en el desarrollo del corazón en embriones de pollo. A tal objeto, los embriones fueron intervenidos durante el estadío 16, mediante la ligadura de la vena vitelina derecha en unos casos, e izquierda en otros, observando sus efectos en embriones de los estadíos 36 y 37 mediante técnicas ultraestructurales de barrido. Nuestros resultados indican que la disminución del volumen sanguíneo causa alteraciones en la morfogénesis cardíaca

Lectin-binding patterns in the development of the cerebellum.

We studied the binding distribution of several lectins, Con A, DBA, UEA and WGA, in the embryonic development of the chick cerebellum between stages 18 to 45 of Hamburger and Hamilton. We observed a differential labeling (in intensity and distribution) in the migratory and cortical layers of the cerebellum anlage with these different lectins. The different distributions and modifications in the labeling pattern suggest intense variations of the glycoproteins and glycosaminoglycans in the extracellular matrix during development. These variations coincide with cellular and organizational phenomena in the migratory and cortical layers, and suggest compartmentalization of the Purkinje cell labeling.

Antigenic relationships between chloroplast and cytosolic fructose-1,6-bisphosphatases.

Cytosolic fructose-1,6-biphosphatases (FBPase, EC 3.1.3.11) from pea (Pisum sativum L. cv Lincoln) and spinach (Spinacia oleracea L. cv Winter Giant) did not cross-react by double immunodiffusion and western blotting with either of the antisera raised against the chloroplast enzyme of both species; similarly, pea and spinach chloroplast FBPases did not react with the spinach cytosolic FBPase antiserum. On the other hand, spinach and pea chloroplast FBPases showed strong cross-reactions against the antisera to chloroplast FBPases, in the same way that the pea and spinach cytosolic enzymes displayed good cross-reactions against the antiserum to spinach cytosolic FBPase. Crude extracts from spinach and pea leaves, as well as the corresponding purified chloroplast enzymes, showed by western blotting only one band (44 and 43 kD, respectively) in reaction with either of the antisera against the chloroplast enzymes. A unique fraction of molecular mass 38 kD appeared when either of the crude extracts or the purified spinach cytosolic FBPase were analyzed against the spinach cytosolic FBPase antiserum. These molecular sizes are in accordance with those reported for the subunits of the photosynthetic and gluconeogenic FBPases. Chloroplast and cytosolic FBPases underwent increasing inactivation when increasing concentrations of chloroplast or cytosolic anti-FBPase immunoglobulin G (IgG), respectively, were added to the reaction mixture. However, inactivations were not observed when the photosynthetic enzyme was incubated with the IgG to cytosolic FBPase, or vice versa. Quantitative results obtained by enzyme-linked immunosorbent assays (ELISA) showed 77% common antigenic determinants between the two chloroplast enzymes when tested against the spinach photosynthetic FBPase antiserum, which shifted to 64% when assayed against the pea antiserum. In contrast, common antigenic determinats between the spinach cytosolic FBPase and the two chloroplast enzymes were less than 10% when the ELISA test was carried out with either of the photosynthetic FBPase antisera, and only 5% when the assay was performed with the antiserum to the spinach cytosolic FBPase. These results were supported by sequencing data: the deduced amino acid sequence of a chloroplast FBPase clone isolated from a pea cDNA library indicated a 39,253 molecular weight protein, with a homology of 85% with the spinach chloroplast FBPase but only 48.5% with the cytosolic enzyme from spinach.

Factors affecting the food intake, growth and protein utilization in the Helix aspersa snail. Protein content of the diet and animal age.

Food intake, growth and protein utilization were studied in the Helix aspersa snail by means of a nutritional balance experiment. This was designed in a 5 x 11 factorial arrangement involving 5 diets of different protein content (10.0, 12.5, 15.0, 17.5 and 20.0%) and 11 animal ages (ranging over 15 to 180 days of age). Dietary protein content and animal age determined food intake and protein utilization. The variations of dry matter and protein intake rates, growth rates, growth efficiency factors and protein retention rates, depending on dietary protein content or animal age were in agreement with what happens in other animals. These results suggest that there is no need to use diets with more than 17.5% of crude protein in these animals.

Protein and energy requirements for maintenance of indigenous Granadina goats.

Sixteen adult castrated male goats of the Granadina breed, with initial live weights ranging from 26.0 to 33.3 kg were used in two experiments to determine their protein and energy requirements for maintenance. Digestibility, nitrogen and energy balance measurements were made during the experiments. Two diets, which were based on pelleted lucerne (Medicago sativa) hay alone or on this forage and barley, were individually given at about maintenance level once daily. Gas exchange was measured using open-circuit respiration chambers. Fasting heat production was also determined. By regression analysis endogenous urinary N and maintenance requirements for N were estimated to be 119 mg/kg body-weight (W)0.75 per d and 409 mg total N/kg W0.75 per d respectively. Fasting heat production was 324 kJ/kg W0.75. The energy requirement for maintenance was calculated by regression of energy balance on metabolizable energy (ME) intake and a value of 443 kJ/kg W0.75 per d was found. The overall efficiency of utilization of ME for maintenance was 0.73.

Protein and energy metabolism of lactating Granadina goats.

Twelve goats of the Granadina breed in mid- and late lactation were used in two consecutive years to determine their protein and energy requirements for lactation. The animals were individually fed on diets based on pelleted lucerne (Medicago sativa) hay and barley. A total of six balance experiments were carried out. Gas exchange was measured using open-circuit respiration chambers. Milk yield ranged widely from 0.649 to 1.742 kg/d in the first year and from 0.222 to 1.989 kg/d in the second year, a steady decline in milk output being observed as lactation progressed. Milk composition remained rather constant during the midstage of lactation, with an average content (/kg milk) for total solids, total nitrogen, fat and gross energy of 149.7 g, 5.39 g, 58.8 g and 3.59 MJ respectively. Total endogenous N, endogenous urinary N and maintenance requirement for N in lactating goats were estimated to be 244, 218 mg N/kg body-weight (W)0.75 per d and 478 mg total N/kg W0.75 per d respectively from regression equations. A constant efficiency of use of dietary N for milk N plus retained N of 51.0% was found. By regressing milk energy plus apparent body energy retention or loss on metabolizable energy (ME) intake, the maintenance energy requirement was estimated to be 401 kJ ME/kg W0.75 per d. When estimating the corrected milk yield as milk energy + (0.84 x negative energy retentio) + (1.05 x positive energy retention), regression analysis indicated that the overall efficiency of use of ME for lactation was 66.7%. Also, from a plot of apparent body energy retention v. milk energy yield, both expressed as a percentage of ME intake above maintenance, the efficiency with which ME was used to promote energy retention in the body during lactation was calculated to be 0.907 times that for milk secretion.

Land snails as experimental animals: a study of the variability and distribution of individual weights in Helix aspersa snails born from the same clutch.

A study was carried out to examine the distribution of individual weights in Helix aspersa snails, the aims being to establish the best estimate of the ponderal growth and also to obtain a model growth curve. Four groups of 20 snails from the same clutch were analysed and kept under experimental conditions from birth up to 6 months. The variability of their individual weights within groups was studied by calculating the coefficients of variation every 15 days. At the same time, the assumed normal distribution of those weights was being tested. The coefficients of variation increased with age and the assumed normal distribution of individual weights had to be rejected. By means of a log transformation of the original data, a model growth curve was constructed, and was used to assess the possibility of estimating age from weight. We finally reached the conclusion that median weight, rather than the mean, would be a better measure of central tendency to use until it is possible to obtain selected populations. The difficulty of estimating age from weight is emphasized.