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.

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.

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.

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.

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.

The ontogeny of the cerebellar fissures in the chick embryo.

We have studied the chronology of appearance and the cortical changes which precede the fissures appearing between stages 34 to 40. In this paper we demonstrate the structural modifications of the cerebellar cortex defined as the anlagen of the fissures. The anlage of a fissure begins in a well-determined place of the cerebellar cortex. It begins with a thickening of the internal cortical cell layer, which finally folds. These modifications precede other similar ones which occur in the overlying external granular layer. On the other hand, these cortical structural modifications precede the transversal projection of the fissures. Chronological order of appearance of the anlagen of the fissures, related to the appearance of fissures on the surface of the cerebellum is also given.

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.

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.

Malformation of the venous sinus in human embryos.

2 human embryos of stages XV and XVII are studied, in which the persistence of the left vitelline vein opening into the venous sinus (sinus venosi cordis) is described.

The morphogenesis of the ventricular flow pathways in man.

This work which is based on the study of eight human embryos of 4,5 to 13,6 mm from the Madrid collection, and also on six plastic reconstructions made by the BORN method, and on experiments on chick embryos (ligature of the left vitelline vein in embryos, 3 days of incubation) enables us to reach the following conclusions: no true rotatiorial movements take place during normogensis. Only ventromedial rotation of the right ventricle is observed in stages 12 to 14. This rotation will place the right ventricle, and therefore the vestibulum pulmonalis in front of the aortic infundibulum. We consider that the initiation of cardiac septation is to a great extent due to hemodynamic. causes. During normogenesis in all cases, from the earliest stages of development the presumptive pulmonary outflow tract is always anterior and to the right of the presumptive aortic tract. We do not agree with the statement of GOOR and EDWARDS and VAN PRAAGH R. and VAN PRAAGH S. (1965), that in some stages the presumptive coni of the aorta and pulmonalis are arranged inversely to the way in which they will arranged later.

The formation, septation and fate of the truncus arteriosus in man.

The results of our studies enable us to draw the following conclusions. The truncus appears in the human embryo, between Stages XII and XIII, as a portion of the aortic sac which invaginates into the interior of the pericardial cavity. Therefore it is an arterial portion which is added to the heart. It lengthens progressively. The sigmoid valves form in the angle between the bulbus cordis and the truncus. Septation of the truncus begins when the sixth arterial arches appear in embryos of 6 to 8 mm. The process is very rapid; commencing in embryos of 6 to 7 mm, it is complete in embryos of 10 to 11 mm, that is to say, during only five days. The septation mechanism is extrinsic. The peribranchial mesenchyma which accompanies the aortic sac in its invagination advances principally on the right inferior part and insinuates itself between the fourth and sixth arterial arches, separating the truncus pulmonalis from a portion of the ascending aorta. An aorticopulmonary communication exists for a certain period prior to fusion of the two blocks of mesenchyma; there is a mesenchymal island. On the contrary, in the bulbus cordis septation is effected by the bulbar ridges. Septation of the truncus, which does not exist in the primitive cardiac tube, occurs prior to that of the bulbus cordis. While septation of the truncus has been already completed in embryos of 10 mm, septation of the bulbus cordis is completed only in embryos of 14 to 16 mm. Therefore the bulbus and the truncus are two portions, different in respect of both structure and septation. There is no continuity between the bulbar ridges and the septation of the truncus. They are separated by the sigmoid valves. This makes it possible to observe independent malformations in the bulbus and in the truncus. In the truncus the mesenchyma passes between the two vessels. They do not have a common septum, and it is for this reason that the surgeon can separate them in the mature heart.

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.