Expanding the reach of probiotics through social enterprises.

The rapid rise in microbiome and probiotic science has led to estimates of product creation and sales exceeding $50 billion within five years. However, many people do not have access to affordable products, and regulatory agencies have stifled progress. The objective of a discussion group at the 2017 meeting of the International Scientific Association for Probiotics and Prebiotics was to identify mechanisms to confer the benefits of probiotics to a larger portion of the world's population. Three initiatives, built around fermented food, were discussed with different methods of targeting populations that face enormous challenges of malnutrition, infectious disease, poverty and violent conflict. As new candidate probiotic strains emerge, and the market diversifies towards more personalised interventions, manufacturing processes will need to evolve. Information dissemination through scientific channels and social media is projected to provide consumers and healthcare providers with rapid access to clinical results, and to identify the nearest location of sites making new and affordable probiotic food and supplements. This rapid translation of science to individual well-being will not only expand the beneficiaries of probiotics, but also fuel new social enterprises and economic business models.

A consideration of biomarkers to be used for evaluation of inflammation in human nutritional studies.

To monitor inflammation in a meaningful way, the markers used must be valid: they must reflect the inflammatory process under study and they must be predictive of future health status. In 2009, the Nutrition and Immunity Task Force of the International Life Sciences Institute, European Branch, organized an expert group to attempt to identify robust and predictive markers, or patterns or clusters of markers, which can be used to assess inflammation in human nutrition studies in the general population. Inflammation is a normal process and there are a number of cells and mediators involved. These markers are involved in, or are produced as a result of, the inflammatory process irrespective of its trigger and its location and are common to all inflammatory situations. Currently, there is no consensus as to which markers of inflammation best represent low-grade inflammation or differentiate between acute and chronic inflammation or between the various phases of inflammatory responses. There are a number of modifying factors that affect the concentration of an inflammatory marker at a given time, including age, diet and body fatness, among others. Measuring the concentration of inflammatory markers in the bloodstream under basal conditions is probably less informative compared with data related to the concentration change in response to a challenge. A number of inflammatory challenges have been described. However, many of these challenges are poorly standardised. Patterns and clusters may be important as robust biomarkers of inflammation. Therefore, it is likely that a combination of multiple inflammatory markers and integrated readouts based upon kinetic analysis following defined challenges will be the most informative biomarker of inflammation.

Inflammatory disease processes and interactions with nutrition.

Inflammation is a stereotypical physiological response to infections and tissue injury; it initiates pathogen killing as well as tissue repair processes and helps to restore homeostasis at infected or damaged sites. Acute inflammatory reactions are usually self-limiting and resolve rapidly, due to the involvement of negative feedback mechanisms. Thus, regulated inflammatory responses are essential to remain healthy and maintain homeostasis. However, inflammatory responses that fail to regulate themselves can become chronic and contribute to the perpetuation and progression of disease. Characteristics typical of chronic inflammatory responses underlying the pathophysiology of several disorders include loss of barrier function, responsiveness to a normally benign stimulus, infiltration of inflammatory cells into compartments where they are not normally found in such high numbers, and overproduction of oxidants, cytokines, chemokines, eicosanoids and matrix metalloproteinases. The levels of these mediators amplify the inflammatory response, are destructive and contribute to the clinical symptoms. Various dietary components including long chain omega-3 fatty acids, antioxidant vitamins, plant flavonoids, prebiotics and probiotics have the potential to modulate predisposition to chronic inflammatory conditions and may have a role in their therapy. These components act through a variety of mechanisms including decreasing inflammatory mediator production through effects on cell signaling and gene expression (omega-3 fatty acids, vitamin E, plant flavonoids), reducing the production of damaging oxidants (vitamin E and other antioxidants), and promoting gut barrier function and anti-inflammatory responses (prebiotics and probiotics). However, in general really strong evidence of benefit to human health through anti-inflammatory actions is lacking for most of these dietary components. Thus, further studies addressing efficacy in humans linked to studies providing greater understanding of the mechanisms of action involved are required.

Clinical evidence for immunomodulatory effects of probiotic bacteria.

Close, tightly orchestrated interactions between the intestinal epithelium and the mucosa-associated immune system are critical for normal intestinal absorptive and immunological functions. Recent data indicate that commensal intestinal microbiota represents a major modulator of intestinal homeostasis. This review analyzes the process of intestinal colonization and the interaction of microbiota with the intestinal epithelium and mucosal immune system, with special reference to the first years of extrauterine life. Dysregulation of the symbiotic interaction between intestinal microbiota and the mucosa may result in a pathological condition with potential clinical repercussions. Based on the concept that there is a beneficial and symbiotic relation between the host and endogenous microbiota, strategies aimed at directly modulating intestinal microbiota with regard to disease prevention or treatment have been developed. One strategy involves administering viable probiotic bacteria. Clinical evidence for the beneficial effect of probiotics in the prevention and/or treatment of necrotizing enterocolitis, infectious and antibiotic-associated diarrhea, allergic diseases, and inflammatory bowel disorders is reviewed herein.

Diversion of cytoskeletal processes by Shigella during invasion of epithelial cells.

Shigella, the causative agent of bacillar dysentery, invades colonic epithelial cells and moves intracellularly to spread from cell to cell. The processes of Shigella entry, determined by the Ipa proteins, and of actin-based motility, dependent on the IcsA/VirG protein, represent different levels of bacterial manipulation of the cell cytoskeleton.

Bacterial signals and cell responses during Shigella entry into epithelial cells.

Shigella invades epithelial cells by inducing cytoskeletal reorganization localized at the site of bacterial-host cell interaction. During entry, the Shigella type III secretion apparatus allows the insertion of a pore that contains the IpaB and IpaC proteins into cell membranes. Insertion of this complex is thought to allow translocation of the carboxy-terminus moiety of IpaC, but also of other Shigella effectors, such as IpaA, into the cell cytosol. IpaC triggers actin polymerization and the formation of filopodial and lamellipodial extensions dependent on the Cdc42 and Rac GTPases. IpaA, on the other hand, binds to the focal adhesion protein vinculin and induces depolymerization of actin filaments. IpaA and the GTPase Rho are not required for actin polymerization at the site of bacterial contact with the cell membrane, but allow the transformation of the IpaC-induced extensions into a structure that is productive for bacterial entry. Rho is required for the recruitment at entry foci of ezrin, a cytoskeletal linker required for Shigella entry, and also of the Src tyrosine kinase. The Src tyrosine kinase activity, which is required for Shigella-induced actin polymerization, also appears to be involved in a negative regulatory loop that downregulates Rho at the site of entry.

Binding of the Shigella protein IpaA to vinculin induces F-actin depolymerization.

Shigella flexneri, the causative agent of bacillary dysentery, enters into epithelial cells by a macropinocytic process. IpaA, a Shigella protein secreted upon cell contact, binds to the focal adhesion protein vinculin and is required for efficient bacterial uptake. IpaA was shown here to bind with high affinity to the N-terminal residues 1-265 of vinculin. Using co-sedimentation and solid-phase assays, we demonstrated that binding of IpaA to vinculin strongly increases the association of vinculin with F-actin. We also characterized a depolymerizing activity on actin filaments associated with the vinculin-IpaA complex both in vitro and in microinjected cells. We propose that the conformational change of vinculin induced by IpaA binding allows interaction of the vinculin-IpaA complex with F-actin and subsequent depolymerization of actin filaments.

Interactions between tissues engaged in wound repair and skeletal muscle regeneration.

Enhanced accumulation of granulation tissue in a wound healing model was observed in the presence of regenerating skeletal muscle. This study characterized the tissue accumulated in the normal wound healing and regeneration-adapted models. DNA, RNA, total protein, collagen, and noncollagen protein content, and protein:DNA and noncollagen protein:collagen ratios were determined using standard methods. DNA did not differ; but RNA increased and total protein decreased compared to early granulation tissue. Mean protein: DNA ratio was lower in regeneration-associated granulation tissue compared to age-matched normal and early granulation tissue. Collagen was reduced and noncollagen protein was increased in tissue surrounding regenerating muscle, yielding an elevated noncollagen:collagen ratio. The relative cellularity of granulation tissue was enhanced and collagen deposition potentially reduced, when accompanied by regeneration. Factors from a regeneration environment can interact with cells engaged in wound repair and potentially alter their behavior.

An in vivo model for evaluating wound repair and regeneration microenvironments.

BACKGROUND: Factors differentially regulating regeneration and repair as contrasting resolutions to injury are unknown. We adapted a common wound healing model to further characterize mammalian wound repair and regeneration microenvironments. METHODS: Polyvinyl alcohol sponges, sponges containing minced muscle, or blocks of minced muscle were implanted onto the backs of Fischer rats. Vascularization was assessed by infusion with india ink and progress of regeneration was evaluated histologically. RESULTS: Regeneration occurring within sponges was histologically similar to that of free muscle blocks; but, was initiated more slowly. Vascularization of minced muscle implanted in sponges was delayed slightly compared to implanted free muscle blocks. CONCLUSIONS: Regeneration of minced muscle in sponges parallels normal regeneration of free minced muscle. Incorporation into a wound repair model provides access to interstitial fluids conditioned by regenerating muscle and will lead to more detailed comparisons of the content and properties of repair and regeneration microenvironments.

Biochemical correlates of dexamethasone-induced relative cardiomegaly in neonatal rats.

Dexamethasone induces a relative cardiomegaly in neonatal rats. Biochemical analyses, performed on hearts at seven days postpartum, disclosed that dexamethasone increased protein content without significantly altering DNA or RNA content. While dexamethasone decreased absolute actinomyosin and collagen content, the relative abundance of these proteins as a function of heart mass was increased and the ratio of muscle protein to collagen remained constant. In addition, dexamethasone increased glycogen but decreased fatty acid content. Thus, dexamethasone treatment of rats during the neonatal period appears to induce a relative hypertrophic cardiomyopathy complicated by fibrosis and metabolic derrangement of myocardial maturation.

Interstitial fluids associated with wound repair support proliferation but not differentiation of neonatal rat myoblasts in vitro.

The ability of wound fluids to support events required for skeletal muscle regeneration was examined. Wound fluids were obtained from polyvinyl alcohol sponges 1, 3, 5, 10, and 15 days after implantation. Neonatal rat L8 myoblasts were used to test the ability of early wound fluids to promote myoblast proliferation and late wound fluids to promote myoblast differentiation-two characteristics deemed critical for effective skeletal muscle regeneration. Early wound fluids (1- and 3-day) stimulated DNA replication by myoblasts, as judged by tritiated thymidine uptake, up to ninefold (p < 0.05). Later wound fluids (5-, 10-, and 15-day) displayed decreasing ability to stimulate proliferation, with 15-day wound fluid failing to significantly stimulate proliferation. In contrast, myoblast differentiation, as judged by myotube fusion and creatine kinase activity, was progressively reduced by wound fluids of increasing age. In fact, late wound fluids (5, 10, and 15 days) reduced myotube fusion by 88% to 100% and depressed creatine kinase activity by 60% to 75% (p < 0.05). Thus, wound fluids from a repair environment appear to support myoblast proliferation early but suppress myoblast differentiation later. These characteristics suggest that the wound repair environment cannot fully support skeletal muscle regeneration.

Dexamethasone-induced histopathology of neonatal rat myocardium.

Dexamethasone-induced relative cardiomegaly in neonatal rats mimics hypertrophic cardiomyopathy seen in premature infants receiving dexamethasone for bronchopulmonary dysplasia. Dexamethasone reduced cellular density by approximately 20%. However, dose related increases in mean fiber diameter were seen and mitotic index was doubled at 12.5 micrograms dexamethasone/injection. In addition, dexamethasone induced collagen accumulation throughout the myocardium in an apparently dose dependent manner. These data suggest that dexamethasone promotes (i) myocardial hypertrophy and possibly hyperplasia and (ii) fibrosis and possibly fibroplasia. Further studies are required to determine relationships between disseminated collagen deposition and other suggested metabolic changes and the observed compensatory myocardial hypertrophy and hyperplasia.

Insulin reduces basal release of atrial natriuretic factor by isolated, perfused neonatal pig hearts.

Plasma atrial natriuretic factor (ANF) levels are markedly altered in diabetic rodents suggesting that insulin might be a modulator of ANF release. Effects of insulin (25 mU/ml) on basal (nonstimulated) immunoreactive (ir)-ANF release by isolated neonatal pig hearts paced at 150 beats/min and perfused with no change in atrial stretch were examined. Release of ir-ANF decreased with age from 1-3 days postpartum (146.0 +/- 38.4 to 62.2 +/- 20.5 fmol/g/min). Insulin stimulated myocardial glucose utilization and lactate production approximately 2-fold, but uniformly decreased ir-ANF release by approximately 25%, regardless of age. Correlations between ir-ANF release and myocardial metabolism suggest independent influences of insulin on these two events. In addition, it appears that this model might be reasonably exploited to investigate episodic ANF release and its regulation during ontogeny and in a variety of physiological states.

Phosphoprotein phosphatase activity in regenerating forelimbs of adult newts, Notophthalmus viridescens.

Traditional signal transduction pathways appear to be operating during amphibian forelimb regeneration, as in other developing systems. A consistent picture appears to be developing that suggests that growth factors promoting proliferation during regeneration act through cAMP-independent mechanisms while factors that modulate differentiation and morphogenesis include signaling pathways in which cAMP participates. Although our understanding of these pathways is far from complete, it is becoming increasingly evident that further characterization of signal transduction events will facilitate (a) identifying major signaling substances, (b) determining the cellular events in regeneration that they modulate, and (c) defining the mechanisms through which they act. The present study demonstrates that PPtases, and in particular PTPases, can be studied in regenerating newt limbs. More importantly, this investigation demonstrates that there are progressive and significant increases in PPtase activity during regeneration. Moreover, the observed patterns of PPtase activity during regeneration conform to an emerging picture that ligands acting through receptors possessing intrinsic tyrosine kinase activity promote proliferation within the regeneration blastema. However, the substrates used in this study precluded acquiring insights about modulating effects of serine/threonine kinases (e.g., PKA and PKC). Nevertheless, these data suggest that activation of PPtases contribute to the orchestration of the diverse cellular activities required to regenerate a vertebrate appendage.

Dexamethasone induces a transient relative cardiomegaly in neonatal rats.

Effects of s.c. administration of dexamethasone (1-25 micrograms/rat pup/d) during the first 5 d postpartum on survival rates, body weight, heart weight, heart-to-body weight ratios, and heart dimensions were determined. Dexamethasone decreased survival, body weight, and heart weight, but increased heart-to-body weight ratios and myocardial percentage of dry weight (0.44 to 0.52-1.06 and 20 to 23%, respectively; p less than 0.05) by 7 d postpartum. By 21 d postpartum, differences in absolute body weight and heart weight between control and experimental animals were reduced; however, myocardial percentage of dry weight and heart-to-body weight ratios were indistinguishable. Microscopic analysis of 7-d-old hearts disclosed that interventricular septum, left ventricular free wall thicknesses, and left ventricle chamber diameter were reduced (0.93 to 0.76-0.85 mm, 1.19-1.34 to 0.92-1.07 mm, and 1.35-1.40 to 0.89-1.23 mm, respectively; p less than 0.05), whereas right ventricular free wall thickness was unaffected and right ventricle chamber diameter was increased (0.29-0.31 to 0.42-0.46 mm) by dexamethasone. Thus, dexamethasone reduced survival and retarded growth of neonatal rats. Retardation of heart growth was less severe, producing a transient relative cardiomegaly characterized by 1) increased myocardial percentage of dry weight, 2) increased left ventricular free wall-to-chamber ratio, and 3) retention of absolute right ventricular free wall thickness. This relative cardiomegaly appeared to be resolved by 21 d postpartum, as reflected in myocardial percentage of dry weight and heart-to-body weight ratios.(ABSTRACT TRUNCATED AT 250 WORDS)

Hypertrophic cardiomyopathy associated with dexamethasone therapy for bronchopulmonary dysplasia.

The potential induction of cardiac effects by high-dose dexamethasone therapy was evaluated prospectively in 13 respirator-dependent infants with bronchopulmonary dysplasia by means of two-dimensional and M-mode echocardiography. The initial divided dose of dexamethasone was 500 micrograms/kg per day, tapered progressively for as long as 6 weeks. Evaluations were made before treatment and at 3, 7, 14, 21, 28, 35, and 42 days after the start of dexamethasone therapy. This regimen was associated with a significant (p less than 0.01) increase in thickness of the interventricular septum (2.60 +/- 0.09 to 4.00 +/- 0.16 mm), diastolic left ventricular free wall (2.80 +/- 0.13 to 4.06 +/- 0.20 mm), and diastolic right ventricular free wall (1.55 +/- 0.08 to 2.02 +/- 0.12 mm). In addition, seven dexamethasone-treated infants but no control infants had systolic anterior motion of the mitral valve (p less than 0.001). These effects were transient, reached their maximal degree by the third week of treatment, and approached pretreatment conditions by the sixth week of treatment. Ejection fraction was not affected; heart rate and mean arterial pressure were transiently increased during dexamethasone therapy. We conclude that a transient absolute myocardial hypertrophy is associated with dexamethasone therapy in infants with bronchopulmonary dysplasia. The mechanism or mechanisms through which this hypertrophy arises and the cardiopulmonary implications are unclear.

Atrial natriuretic peptide in newborn piglets with a patent ductus arteriosus.

Atrial natriuretic peptide (ANP) is a hormone involved in fluid and blood pressure homeostasis. We studied the effects of left-to-right shunting through a patent ductus arteriosus on blood pressure changes and plasma ANP concentrations in newborn piglets. In five experimental piglets, the ductus arteriosus was bathed with PGE1 and infiltrated with formalin to maintain its patency. In four age-matched control piglets, the ductus arteriosus was ligated. Plasma ANP concentrations and blood pressure determinations were obtained prior to (base-line) and 25 +/- 1 h (day 1), and 48 +/- 1 h (day 2) after surgery. Radionuclide-microsphere determinations of left-to-right patent ductus arteriosus shunts were performed on days 1 and 2 in the 5 piglets with a patent ductus arteriosus. Plasma ANP concentrations were significantly elevated in the left atrium on day 1 and the right atrium on day 2 in the PDA piglets. No correlation was demonstrated between plasma ANP concentrations and right or left atrial pressures. We conclude that left and right plasma atrial ANP concentrations are significantly elevated in newborn piglets with left-to-right patent ductus arteriosus shunts.