Gut microbiota: next frontier in understanding human health and development of biotherapeutics

The gut microbiota is a remarkable asset for human health. As a key element in the development and prevention of specific diseases, its study has yielded a new field of promising biotherapeutics. This review provides comprehensive and updated knowledge of the human gut microbiota, its implications in health and disease, and the potentials and limitations of its modification by currently available biotherapeutics to treat, prevent and/or restore human health, and future directions. Homeostasis of the gut microbiota maintains various functions which are vital to the maintenance of human health. Disruption of the intestinal ecosystem equilibrium (gut dysbiosis) is associated with a plethora of human diseases, including autoimmune and allergic diseases, colorectal cancer, metabolic diseases, and bacterial infections. Relevant underlying mechanisms by which specific intestinal bacteria populations might trigger the development of disease in susceptible hosts are being explored across the globe. Beneficial modulation of the gut microbiota using biotherapeutics, such as prebiotics, probiotics, and antibiotics, may favor health-promoting populations of bacteria and can be exploited in development of biotherapeutics. Other technologies, such as development of human gut models, bacterial screening, and delivery formulations eg, microencapsulated probiotics, may contribute significantly in the near future. Therefore, the human gut microbiota is a legitimate therapeutic target to treat and/or prevent various diseases. Development of a clear understanding of the technologies needed to exploit the gut microbiota is urgently required

Effect of Orally Administered Microencapsulated FA-Producing L. fermentum on Markers of Metabolic Syndrome: An In Vivo Analysis

Ferulic Acid (FA) is a natural phenolic acid produced by a number of lactic acid bacteria. FA has a number of beneficial properties, including: antioxidant activity, anti-tumorigenic properties and cholesterol-lowering capabilities. Our group has previously screened lactobacilli for FA production, and selected L. fermentum ATCC 11976 (L.f. 11976) as one of the best producers. Alginate-polylysine-alginate (APA) microencapsulation has proven successful for the oral delivery of this strain to the colon, where production of FA is greatest. The aim of this study was to investigate the role of APA microencapsulated L.f. 11976 to modulate markers of metabolic syndrome. The antioxidant activity, as a potential mechanism of action to treat/prevent metabolic syndrome of free and microencapsulated L.f. 11976 was quantified. A high-fat fed BioF1B Golden Syrian hamster model was used to investigate the effects of orally administered microencapsulated L.f. 11976 on markers of metabolic syndrome. Results demonstrate that the microencapsulated L.f. 11976 formulation greatly reduced the adiposity index (p = 0.0014), serum insulin (p = 0.0042), insulin resistance (p = 0.0096), glycosylated albumin (p = 0.00013), serum leptin (p = 0.048), serum uric acid (p =0.025) serum total cholesterol (p = 0.024), serum esterified cholesterol (p = 0.0328) and free non-esterified fatty acid (p = 0.029) levels in the treated animals. This research indicates that the probiotic L.f. 11976 microencapsulated formulation may significantly delay the onset of insulin resistance, hyperglycemia, hyperinsulinemia, dyslipidemia and obesity, indicating a lower risk of diabetes and cardiovascular disease. We propose and discuss the potential mechanism(s) of action by which FA is acting. With these in mind, further in vivo studies are required to validate the therapeutic effects of the formulation and to investigate the mechanism(s) of action by which the probiotic formulation is acting

Oral Probiotic Microcapsule Formulation Ameliorates Non-Alcoholic Fatty Liver Disease in Bio F1B Golden Syrian Hamsters

The beneficial effect of a microencapsulated feruloyl esterase producing Lactobacillus fermentum ATCC 11976 formulation for use in non-alcoholic fatty liver disease (NAFLD) was investigated. For which Bio F1B Golden Syrian hamsters were fed a methionine deficient/choline devoid diet to induce non-alcoholic fatty liver disease. Results, for the first time, show significant clinical benefits in experimental animals. Examination of lipids show that concentrations of hepatic free cholesterol, esterified cholesterol, triglycerides and phospholipids were significantly lowered in treated animals. In addition, serum total cholesterol, triglycerides, uric acid and insulin resistance were found to decrease in treated animals. Liver histology evaluations showed reduced fat deposits. Western blot analysis shows significant differences in expression levels of key liver enzymes in treated animals. In conclusion, these findings suggest the excellent potential of using an oral probiotic formulation to ameliorate NAFLD

' Lactobacillus fermentum ' 3872 genome sequencing reveals plasmid and chromosomal genes potentially involved in a probiotic activity.

In this report we describe a ' Lactobacillus fermentum ' 3872 plasmid (pLF3872) not previously found in any other strain of this species. The analysis of the complete sequence of this plasmid revealed the presence of a gene encoding a large collagen binding protein (CBP), as well as the genes responsible for plasmid maintenance and conjugation. Potential roles of CBP and a chromosomally encoded fibronectin-binding protein (FbpA) in probiotic activity are discussed

Driven Assembly of Lignin into Microcapsules for Storage and Delivery of Hydrophobic Molecules

Oil-filled microcapsules of kraft lignin were synthe- sized by first creating an oil in water emulsion followed by a high- intensity, ultrasound-assisted cross-linking of lignin at the water/oil interface. The rationale behind our approach is based on promoting documented lignin hydrophobic interactions within the oil phase, followed by locking the resulting spherical microsystems by covalent cross-linking using a high intensity ultrasound treatment. As further evidence in support of our rationale, confocal and optical microscopies demonstrated the uniformly spherical morphology of the created lignin microparticles. The detailed elucidation of the cross-linking processes was carried out using gel permeation chromatography (GPC) and quantitative 31P NMR analyses. The ability of lignin microcapsules to incorporate and release Coumarin-6 was evaluated in detail. In vitro studies and confocal laser scanning microscopy analysis were carried out to assess the internalization of capsules into Chinese hamster ovary (CHO) cells. This part of our work demonstrated that the lignin microcapsules are not cytotoxic and readily incorporated in the CHO cells

Microencapsulated ferulic acid esterase active «Lactobacillus fermentum» for the reduction of inflammation and cholesterol in metabolic syndrome

Metabolic syndrome (MetS) is an important public health concern of industrialized countries. MetS is a cluster of metabolic disturbances, including abdominal obesity, atherogenic dyslipidemia, elevated blood pressure, insulin resistance and the presence of a proinflammatory and prothrombotic state. Currently, there is no successful therapeutic intervention for the prevention and treatment of this disorder. Chronic systemic inflammation and hypercholesterolemia are two important therapeutic targets of MetS. The goal of this work is to develop a novel microencapsulated ferulic acid-producing probiotic Lactobacillus formulation as a MetS biotherapeutic. Specifically, a number of selected probiotic strains were screened for their ferulic acid esterase activity to select the best probiotic producer of ferulic acid from ethyl ferulate, a natural substrate. Ferulic acid is a molecule with important therapeutic properties relevant to the development of a MetS biotherapeutic. Due to the harsh conditions of the gastrointestinal tract, we propose microencapsulation as a carrier capable of protecting the viability and activity of the probiotic cells. Hence, the best ferulic acid-producing L. fermentum was microencapsulated using alginate-polylysine-alginate microcapsules. The probiotic viability and activity in the microcapsules was significantly higher than that of non-microencapsulated probiotic cells following exposure to gastrointestinal conditions in an in vitro model, demonstrating its suitability for oral delivery. The probiotic L. fermentum formulation was shown to possess anti-inflammatory properties, in vitro. Specifically, the probiotic reduced the secretion of macrophage pro-inflammatory cytokines and provided protection of the intestinal epithelial integrity in a co-culture model of the epithelium. As well, the probiotic formulation demonstrated important cholesterol-lowering activity, by assimilating cholesterol and via inhibition of colon epithelium cholesterol uptake, determined in vitro. Results also demonstrate that L. fermentum, administered in animal models of MetS, significantly reduced a number of MetS-associated risk factors, including cholesterol, adiposity, insulin resistance and hyperglycemia. This novel work indicates the potential of a ferulic acid-producing microencapsulated probiotic formulation as a biotherapeutic for MetS treatment and prevention. Further studies, including clinical investigations, are required to demonstrate the full potential of this probiotic biotherapeutic for the management of MetS.Le syndrome métabolique (SMet) est une préoccupation importante des pays industrialisés au niveau de la santé publique. SMet est un groupe de troubles métaboliques, qui comprend l'obésité abdominale, la dyslipidémie athérogènique, l'hypertension artérielle, la résistance à l'insuline et la présence d'un état pro-inflammatoire et pro-thrombotique. Actuellement, il n'existe aucune intervention thérapeutique efficace pour la prévention et le traitement de ce syndrome. L'inflammation systémique chronique et l'hypercholestérolémie sont deux cibles thérapeutiques importantes du SMet. Le but de cette recherche est de développer une nouvelle formulation biothérapeutique de microcapsules probiotiques produisant de l'acide férulique pour combattre le SMet. En particulier, des souches probiotiques ont été évaluées pour leur activité d'estérase d'acide férulique pour choisir le probiotique produisant le plus d'acide férulique. L'acide férulique est une molécule possédant des propriétés thérapeutiques importantes relatives au développement d'une biothérapeutique SMet. La souche de L. fermentum sélectionnée a été microencapsulée à l'intérieur de microcapsules d'alginate-polylysine-alginate. La viabilité des cellules probiotiques et leur activité dans les microcapsules était significativement supérieure à celle des cellules probiotiques non-microencapsulées une fois soumises à des conditions gastro-intestinales dans un modèle in vitro, ce qui démontre leur aptitude à l'administration orale. La formulation probiotique L. fermentum a démontré des propriétés anti-inflammatoires, in vitro. En particulier, le probiotique L. fermentum réduit la sécrétion de cytokines pro-inflammatoires par les macrophages et assure la protection de l'intégrité épithéliale intestinale dans un modèle in vitro de l'épithélium. La formulation probiotique a aussi démontré une activité anti-cholestérol importante en assimilant le cholestérol et par l'inhibition de l'absorption du cholestérol par l'épithélium du côlon, déterminée in vitro. Les résultats démontrent également que L. fermentum, administré dans des modèles animaux de SMet, réduit de manière significative plusieurs facteurs associés au SMet, y compris le cholestérol, l'adiposité, l'insulinorésistance et l'hyperglycémie. Cette nouvelle recherche démontre le potentiel d'une formulation probiotique microencapsulée, produisant de l'acide férulique, comme un biothérapeutique pour le traitement et la prévention du SMet. D'autres études, y compris des études cliniques, sont nécessaires pour démontrer la pertinence de ce produit