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Gut flora consists of microorganisms that live in the digestive tracts and is the largest reservoir of human flora. The human body, consists of about 100 trillion cells, carries about ten times as many microorganisms in the intestines. Bacteria make up most of the flora in the colon and up to 60% of the dry mass of feces, between 300 and 1000 different species live in the gut. The microorganisms perform a host of useful functions, such as fermenting unused energy substrates, training the immune system, preventing growth of harmful, pathogenic bacteria, regulating the development of the gut, producing vitamins for the host (such as biotin and vitamin K), and producing hormones to direct the host to store fats. Multiple Sclerosis (MS) is an inflammatory disease in which the fatty myelin sheaths around the axons of the brain and spinal cord are damaged, leading to demyelination and scarring as well as a broad spectrum of signs and symptoms. Disease onset usually occurs in young adults, and it is more common in women. Researchers at the Max Planck Institute of Neurobiology in Munich, Germany have found an astonishing evidence that suggests MS is triggered by natural intestinal flora, the so-called friendly bacteria that reside in the gut. They discovered this by allowing some of the genetically modified mice to continue with their normal gut bacteria intact, while removing the intestinal flora in the others and keeping them under sterile conditions.The mice that kept their gut bacteria developed MS-like symptoms.
But the mice that had their gut bacteria removed remained healthy, despite their genetic predisposition to MS. However, when they then inoculated these mice with normal gut flora, their T-cells and B-cells increased, as did their cytokine and antibody production, and they eventually developed symptoms and fell ill. The team now wants to investigate the complete microbial genomes of people with MS and compare them to people without MS.
Science, Technology and Medicine open access publisher.Publish, read and share novel research. Lactic Acid Bacteria as Probiotics:Characteristics, Selection Criteria and Role in Immunomodulation of Human GI Muccosal BarrierDaoud Harzallah and Hani Belhadj[1] Laboratory of Applied Microbiology, Faculty of Natural and Life Sciences, University Ferhat Abbas, Setif, Algeria1.
Miso soup is made from fermented soybeans, sea salt, koji and some blends are made with barley, brown rice, buckwheat and white rice.
Eating miso soup can help to reduce your risk for prostate, breast, colon and lung cancers. This 500lb Guy Thought He Was Content With His Life… Until He Started Practicing Yoga. The metabolic activities performed by these bacteria resemble those of an organ. It is estimated that these gut flora have around 100 times as many genes as there are in the human genome. However, in certain conditions, some species are thought to be capable of causing disease by producing infection or increasing cancer risk for the host. MS affects the ability of nerve cells in the brain and spinal cord to communicate with each other effectively. They say the bacteria first activated the immune T-cells, then the B-cells, which resulted in an attack on the myelin layer in the brain, developing brain inflammation similar to MS.
They also had fewer T-cells in their gut, their spleens produced fewer inflammatory substances like cytokines, and their B-cells produced few if any antibodies against myelin. Together with the surface proteins of the myelin layer, these then stimulate the B cells to form pathogenic antibodies.
IntroductionAs it was reported by Chow (2002), the notion that food could serve as medicine was first conceived thousands of years ago by the Greek philosopher and father of medicine, Hippocrates, who once wrote: 'Let food be thy medicine, and let medicine be thy food'. Epithelial cells lining the gastrointestinal tract are able to respond to infection by initiating either nonspecific or specific host-defence response (Kagnoff and Eckmann 1997, Strober 1998).
The harmful species under this group when occurring at the wrong place can make you fall ill at random. It’s full of beneficial bacteria, enzymes, protein, vitamin B12, B2, vitamin K, vitamin E, tryptophan, choline, lecithin and linoleic acid. In 1945, when the atomic bomb was dropped in Nagasaki, Japan, patients and staff at the St. Most likely MS occurs as a result of some combination of genetic, environmental and infectious factors, and possibly other factors like vascular problems. Potential and established health benefits associated with the usage of probiotics (Leroy et al., 2008).
However, during recent times, the concept of food having medicinal value has been reborn as 'functional foods'.
Bacterial adhesion to the host cell or recognition by the host cell is often an essential first stage in the disease process. Key and desirable criteria for the selection of probiotics in commercial applications (Vasiljevic and Shah, 2008). The list of health benefits accredited to functional food continues to increase, and the gut is an obvious target for the development of functional foods, because it acts as an interface between the diet and all other body functions.
A wide range of gastrointestinal cell surface constituents, such as several glygoconjucates, can serve as receptors for bacterial adherence (Servin and Coconnier 2003, Pretzer et al., 2005). Some of them are even discerned in the gut flora and they can cause much discomfort in case of human beings. They all managed to survive and stay healthy, while others who were the same distance or even farther away suffered from leukemia and radiation burns. One of the most promising areas for the development of functional food components lies in the use of probiotics and prebiotics which scientific researches have demonstrated therapeutic evidence.
Furthermore, epithelial cells express constitutively host pattern recognition receptors (PRRS), such as Toll-like receptors (TLR). Under the supervision of Shinichiro Akizuki, M.D, the patients and staff ate a daily diet of miso soup, brown rice, vegetables and seaweed. Nowadays, consumers are aware of the link among lifestyle, diet and good health, which explains the emerging demand for products that are able to enhance health beyond providing basic nutrition. TLRs are also found on innate immune cells, such as dendritic cells and macrophages (Vinderola et al., 2005). They can be found in fermented products as meat, milk products, vegetables, beverages and bakery products. They are part of the microbiota on mucous membranes, such as the intestines, mouth, skin, urinary and genital organs of both humans and animals, and may have a beneficial influence on these ecosystems. Other known recognition receptors are nucleotide-binding oligomerization domain proteins, which recognize both gram-positive and gram-negative bacteria. LAB that grow as the adventitious microflora of foods or that are added to foods as cultures are generally considered to be harmless or even an advantage for human health. Since their discovery, LAB has been gained mush interest in various applications, as starter cultures in food and feed fermentations, pharmaceuticals, probiotics and as biological control agents. Increased epithelial barrier permeability is frequently associated with gastrointestinal disorders contributing to both disease onset and persistence (Lu and Walker 2001, Berkes 2003). In food industry, LAB are widely used as starters to achieve favorable changes in texture, aroma, flavor and acidity (Leory and De Vuyst, 2004). The gatekeeper of the paracellular pathway is the tight junction, which is an apically located cell-cell junction between epithelial cells. The tight junction permits the passage of small molecules, such as ions, while restricting the movement of large molecules, such as antigens and microorganisms, which can cause inflammation. Du to their antimicrobial and antioxidant activities some LAB strains are used in food biopreservation. Origine and safety of probiotics An old dogma of probiotic selection has been that the probiotic strains should be of “human origin”. Many of the indications for probiotic activity have been obtained from effects observed in various clinical situations.
One may argue that from evolutionary point of view, describing bacteria to be of human origin does not make much sense at all. The requirement for probiotics to be of human origin relates actually to the isolation of the strain rather than the “origin” itself.
Usually, the strains claimed to be “of human origin” have been isolated from faecal samples of healthy human subjects, and have therefore been considered to be “part of normal healthy human gut microbiota”. Overview of probioticsThe most tried and tested manner in which the gut microbiota composition may be influenced is through the use of live microbial dietary additions, as probiotics. In reality the recovery of a strain from a faecal sample does not necessarily mean that this strain is part of the normal microbiota of this individual, since microbes passing the GI tract transiently can also be recovered from the faecal samples (Forssten et al., 2011).
In practice it is impossible to know the actual origin of the probiotic strains, regardless of whether they have been isolated from faecal samples, fermented dairy products or any other source for that matter.
Isolation of a strain from faeces of a healthy individual is also not a guarantee of the safety of the strain—such a sample will also always contain commensal microbes which can act as opportunistic pathogens, or even low levels of true pathogens, which are present in the individual at sub-clinical levels. However, at the beginning of this century probiotics were first put onto a scientific basis by the work of Metchnikoff (1908).


He hypothesised that the normal gut microflora could exert adverse effects on the host and that consumption of ‘soured milks’ reversed this effect. However, many species of the genera Lactobacillus, Leuconostoc, Pediococcus, Enterococcus, and Bifidobacterium were isolated frequently from various types of infective lesions.
The origin of the first use can be traced back to Kollath (1953), who used it to describe the restoration of the health of malnourished patients by different organic and inorganic supplements. Later, Vergin (1954) proposed that the microbial imbalance in the body caused by antibiotic treatment could have been restored by a probiotic rich diet; a suggestion cited by many as the first reference to probiotics as they are defined nowadays.
Similarly, Kolb recognized detrimental effects of antibiotic therapy and proposed the prevention by probiotics (Vasiljevic and Shah, 2008) Later on, Lilly and Stillwell (1965) defined probiotics as “…microorganisms promoting the growth of other microorganisms”. The idea of health-promoting effects of LAB is by no means new, as Metchnikoff proposed that lactobacilli may fight against intestinal putrefaction and contribute to long life.
Although minor side effects of the use of probiotics have been reported, infections with probiotic bacteria occur and invariably only in immunocompromised patients or those with intestinal bleeding (Leroy et al., 2008).
Other definitions advanced through the years have been restrictive by specification of mechanisms, site of action, delivery format, method, or host.
An issue of concern regarding the use of probiotics is the presence of chromosomal, transposon, or plasmid-located antibiotic resistance genes amongst the probiotic microorganisms. At this moment, insufficient information is available on situations in which these genetic elements could be mobilised, and it is not known if situations could arise where this would become a clinical problem (Leroy et al., 2008). The mechanism of action of probiotics (e.g, having an impact on the intestinal microbiota or enhancing immune function) was dropped from the definition to encompass health effects due to novel mechanisms and to allow application of the term before the mechanism is confirmed. Furthermore, certain mechanisms of action (such as delivery of certain enzymes to the intestine) may not require live cells.
In vitro safety screenings of probiotics may include, among others, antibiotic resistance assays, screenings for virulence factors, resistance to host defence mechanisms and induction of haemolysis. In relation to food, probiotics are considered as “viable preparations in foods or dietary supplements to improve the health of humans and animals”. According to these definitions, an impressive number of microbial species are considered as probiotics. ConclusionThe individual diversity of the intestinal microflora underscores the difficulty of identifying the entire human microbiota and poses barriers to this ?eld of research.
Selection of probioticsMany in vitro tests are performed when screening for potential probiotic strains. The first step in the selection of a probiotic LAB strain is the determination of its taxonomic classification, which may give an indication of the origin, habitat and physiology of the strain. It is also apparent that even a single strain of probiotic may exert its actions via multiple, concomitant pathways.
All these characteristics have important consequences on the selection of the novel strains (Morelli, 2007). Probiotics have long been used as an alternative to traditional medicine with the goal of maintaining enteric homeostasis and preventing disease. This conclusion was brought forward due to uncertainty of the origin of the human intestinal microflora since the infants are borne with virtually sterile intestine.
Clinical trials have shown that probiotic treatment can reduce the risk of some diseases, especially antibiotic-associated diarrhea, but conclusive evidence is impeded owing to the wide range of doses and strains of bacteria used. However, the panel also underlined a need for improvement of in vitro tests to predict the performance of probiotics in humans.
While many probiotics meet criteria such as acid and bile resistance and survival during gastrointestinal transit, an ideal probiotic strain remains to be identified for any given indication. Many studies, as discussed above, have shown that probiotics increase barrier function in terms of increased mucus, antimicrobial peptides, and sIgA production, competitive adherence for pathogens, and increased TJ integrity of epithelial cells.
Furthermore, it seems unlikely that a single probiotic will be equally suited to all indications; selection of strains for disease-specific indications will be required (Shanahan, 2003). Current investigation into the mechanism of action of speci?c probiotics has focused on probiotic-induced changes in the innate immune functions involvingTLRs and its downstream systems Like NF-?B, and other pathways (Yoon and Sun, 2011).
Although the immunomodulatory effects of probiotics have been demonstrated in experimental animal models of allergy, autoimmunity, and IBD, information from clinical trials in humans is scarce. The ability to adhere to the intestinal mucosa is one of the more important selection criteria for probiotics because adhesion to the intestinal mucosa is considered to be a prerequisite for colonization (Tuomola et al., 2001).
The table below (Table 2) indicates key creteria for sellecting probiotic candidat for commercial application, and figure 1 presents major and cardinal steps for sellecting probiotic candidats.It is of high importance that the probiotic strain can survive the location where it is presumed to be active.
Therefore, more research, especially in the form of well-designed clinical trials, is needed to evaluate the ef?cacy and safety of probiotics (Ezendam and Van Loveren, 2008). For a longer and perhaps higher activity, it is necessary that the strain can proliferate and colonise at this specific location. Probably only host-specific microbial strains are able to compete with the indigenous microflora and to colonise the niches. Besides, the probiotic strain must be tolerated by the immune system and not provoke the formation of antibodies against the probiotic strain. On the other hand, the probiotic strain can act as an adjuvant and stimulate the immune system against pathogenic microorganisms. Basic initial characterization of strain identity and taxonomy should be conducted, followed by evaluation with validated assays both in studies of animal models and in controlled studies in the target host. In vitro assays are frequently conducted that have not been proved to be predictive of in vivo function. Technological robustness must also be determined, such as the strain’s ability to be grown to high numbers, concentrated, stabilized, and incorporated into a ?nal product with good sensory properties, if applicable, and to be stable, both physiologically and genetically, through the end of the shelf life of the product and at the active site in the host.
Assessment of stability can also be a challenge, since factors such as chain length and injury may challenge the typical assessment of colony-forming units, as well as in vivo function (Sanders, 2008).
Dose levels of probiotics should be based on levels found to be ef?cacious in human studies. Furthermore, the impact of product format on Figure 1.Scheme of the Guidelines for the Evaluation of Probiotics for Food Use.
The common quality-control parameter of colony-forming units per gram may not be the only parameter indicative of the ef?cacy of the ?nal product.
Other factors, such as probiotic growth during product manufacture, coating, preservation technology, metabolic state of the probiotic, and the presence of other functional ingredients in the ?nal product, may play a role in the effectiveness of a product. Potential mechanisms of action of probioticsA wide variety of potential beneficial health effects have been attributed to probiotics (Table 3). Claimed effects range from the alleviation of constipation to the prevention of major life-threatening diseases such as inflammatory bowel disease, cancer, and cardiovascular incidents. Some of these claims, such as the effects of probiotics on the shortening of intestinal transit time or the relief from lactose maldigestion, are considered well-established, while others, such as cancer prevention or the effect on blood cholesterol levels, need further scientific backup (Leroy et al., 2008). The mechanisms of action may vary from one probiotic strain to another and are, in most cases, probably a combination of activities, thus making the investigation of the responsible mechanisms a very difficult and complex task.
In general, three levels of action can be distinguished: probiotics can influence human Probiotic organisms can provide a beneficial effect on intestinal epithelial cells in numerous ways.
Gut microbiotaThe human gastrointestinal tract is inhabited by a complex and dynamic population of around 500-1000 of different microbial species which remain in a complex equilibrium. It has been estimated that bacteria account for 35–50% of the volume content of the human colon.
These include Bacteroides, Lactobacillus, Clostridium, Fusobacterium, Bifidobacterium, Eubacterium, Peptococcus, Peptostreptococcus, Escherichia and Veillonella. The bacterial strains with identified beneficial properties include mainly Bifidobacterium and Lactobacillus species. The dominant microbial composition of the intestine have been shown to be stable over time during adulthood, and the microbial patterns are unique for each individual.
However, there are numerous external factors that have potential to influence the microbial composition in the gut as host genetics, birth delivery mode, diet, age, antibiotic treatments and also, other microorganisms as probiotics. The intestine is one of the main surfaces of contact with exogenous agents (viruses, bacteria, allergens) in the human body. It has a primary role in the host defense against external aggressions by means of the intestinal mucosa, the local immune system, and the interactions with the intestinal microbiota (resident and in transitbacteria).
Gut microbiota influences human health through an impact on the gut defense barrier, immune function, nutrient utilization and potentially by direct signaling with the gastrointestinal epithelium (Collado et al., 2009).


In healthy adults, 80% of phylotypes belong to four major phylogenetic groups, which are the Clostiridium leptum, Clostridium coccoides, Bacteroides and Bifidobacteria groups. Also, studies have found that mucosal microbiota is stable along the distal gastrointestinal tract from ileum to rectum, but mucosa-associated microbiota is different from fecal microbiota.
The number of bacterial cells present in the mammalian gut shows a continuum that goes from 101 to 103 bacteria per gram of contents in the stomach and duodenum, progressing to 104 to 107 bacteria per gram in the jejunum and ileum and culminating in 1011 to 1012 cells per gram in the colon (Figure 3a). In addition to the longitudinal heterogeneity displayed by the intestinal microbiota, there is also a great deal of latitudinal variation in the microbiota composition (Figure 3b). The intestinal epithelium is separated from the lumen by a thick and physicochemically complex mucus layer. The microbiota present in the intestinal lumen differs significantly from the microbiota attached and embedded in this mucus layer as well as the microbiota present in the immediate a: variations in microbial numbers and composition across the length of the gastrointestinal tract.
For instance, Bacteroides, Bifidobacterium, Streptococcus, members of Enterobacteriacea, Enterococcus, Clostridium, Lactobacillus, and Ruminococcus were all found in feces, whereas only Clostridium, Lactobacillus, and Enterococcus were detected in the mucus layer and epithelial crypts of the small intestine (Sekirov et al., 2010).
Upon passage through the birth canal, infants are exposed to a complex microbial population.
After the initial establishment of the intestinal microbiota and during the first year of life, the microbial composition of the mammalian intestine is relatively simple and varies widely between different individuals and also with time. Survival and antagonism effects of probiotics in the gutThe intestinal epithelium is the largest mucosal surface in the human body, provides an interface between the external environment and the host. The gut epithelium is constantly exposed to foreign microbes and antigens derived from digested foods. Thus, the gut epithelium acts as a physical barrier against microbial invaders and is equipped with various elements of the innate defense system. In the gut, two key elements govern the interplay between environmental triggers and the host: intestinal permeability and intestinal mucosal defense. Resident bacteria can interact with pathogenic microorganisms and external antigens to protect the gut using various strategies.According to the generally accepted de?nition of a probiotic, the probiotic microorganism should be viable at the time of ingestion to confer a health bene?t. Although not explicitly stated, this de?nition implies that a probiotic should survive GI tract passage and, colonize the host epithelium.
A variety of traits are believed to be relevant for surviving GI tract passage, the most important of which is tolerance both to the highly acidic conditions present in the stomach and to concentrations of bile salts found in the small intestine.
These properties have consequently become important selection criteria for new probiotic functionality. One of the mechanisms by which the gut ?ora resists colonization by pathogenic bacteria is by the production of a physiologically restrictive environment, with respect to pH, redox potential, and hydrogen sul?de production. Probiotic bacteria decrease the luminal pH, as has been demonstrated in patients with ulcerative colitis (UC) following ingestion of the probiotic preparation VSL#3.
Several bacteriocins produced by different species from the genus Lactobacillus have been described. The inhibitory activity of these bacteriocins varies; some inhibit taxonomically related Gram-positive bacteria, and some are active against a much wider range of Gram-positive and Gram-negative bacteria as well as yeasts and molds. Lacticin 3147, a broad-spectrum bacteriocin produced by Lactococcus lactis, inhibits a range of genetically distinct Clostridium dif?cile isolates from healthy subjects and patients with IBD. A further example is the antimicrobial effect of Lactobacillus species on Helicobacter pylori infection of gastric mucosa, achieved by the release of bacteriocins and the ability to decrease adherence of this pathogen to epithelial cells (Gotteland et al., 2006). The pretreatment of intestinal (T84) cells with lactic acid-producing bacteria reduced the ability of pathogenic E. Adhesion and invasion of an intestinal epithelial cell line (Intestine 407) by adherent invasive E.
Probiotics and the mucous layerMost mucosal surfaces are covered by a hydrated gel formed by mucins. Mucins are secreted by specialized epithelial cells, such as gastric foveolar mucous cells and intestinal goblet cells, Goblet cells are found along the entire length of the intestinal tract, as well as other mucosal surfaces.
Of the 18 mucin-type glycoproteins expressed by humans, MUC2 is the predominant glycoprotein found in the small and large bowel mucus. The NH2- and COOH-termini are not glycosylated to the same extent, but are rich in cysteine residues that form intra- and inter-molecular disul?de bonds. These glycan groups confer proteolytic resistance and hydrophilicity to the mucins, whereas the disul?de linkages form a matrix of glycoproteins that is the backbone of the mucous layer (Ohland and MacNaughton, 2010). Although small molecules pass through the heavily glycosylated mucus layer with relative ease, bulk fluid flow is limited and thereby contributes to the development of an unstirred layer of fluid at the epithelial cell surface.
As the unstirred layer is protected from convective mixing forces, the diffusion of ions and small solutes is slowed (Turner, 2009). This gel layer provides protection by shielding the epithelium from potentially harmful antigens and molecules including bacteria from directly contacting the epithelial cell layer, while acting as a lubricant for intestinal motility. Mucins can also bind the epithelial cell surface carbohydrates and form the bottom layer, which is ?rmly attached to the mucosa, whereas the upper layer is loosely adherent. The mucus is the ?rst barrier that intestinal bacteria meet, and pathogens must penetrate it to reach the epithelial cells during infection (Ohland and MacNaughton, 2010).Probiotics may promote mucus secretion as one mechanism to improve barrier function and exclusion of pathogens.
In support of this concept, probiotics have been shown to increase mucin expression in vitro, contributing to barrier function and exclusion of pathogens. Several studies showed that increased mucin expression in the human intestinal cell lines Caco-2 (MUC2) and HT29 (MUC2 and 3), thus blocking pathogenic E.
However, healthy rats did not display increased colonic TFF3 expression after stimulation by VSL#3 probiotics (Caballero-Franco et al., 2007).
Furthermore, mice treated with 1% dextran sodium sulfate (DSS) to induce chronic colitis did not exhibit increased TFF3 expression or wound healing when subsequently treated with VSL#3. This observation indicates that probiotics do not enhance barrier function by up-regulation of TFF3, nor are they effective at healing established in?ammation. Therefore, use of current probiotics is likely to be effective only in preventing in?ammation as shown by studies in animal models (Ohland and MacNaughton, 2010).5.
Interaction of probiotic bacteria with gut epitheliumThe composition of the commensal gut microbiota is probably influenced by the combination of food practices and other factors like the geographical localization, various levels of hygiene or various climates.
The establishment of a normal microbiota provides the most substantial antigenic challenge to the immune system, thus helping the gut associated lymphoid tissus (GALT) maturation. The intestinal microbiota contributes to the anti-inflammatory character of the intestinal immune system. Several immunoregulatory mechanisms, including regulatory cells, cytokines, apoptosis among others, participate in the control of immune responses by preventing the pathological processes associated with excessive reactivity. An interesting premise for probiotic physiological action is their capacity to modulate the immune system. Consequently, many studies have focused on the effects of probiotics on diverse aspects of the immune response. Following consumption of probiotic products, the interaction of these bacteria with intestinal enterocytes initiates a host response, since intestinal cells produce various immunomodulatory molecules when stimulated by bacteria (Delcenseri et al., 2009). Furthermore, The indigenous microbiota is a natural resistance factor against potential pathogenic microorganisms and provides colonization resistance, also known as gut barrier, by controlling the growth of opportunistic microorganisms. It has been suggested that commensal bacteria protect their host against microbial pathogens by interfering with their adhesion and toxic effects (Myllyluoma, 2007).A fraction of ingested probiotics are able to interact with intestinal epithelial cells (IECs) and dendritic cells (DCs), depending on the presence of a dynamic mucus layer.
Probiotics can occasionally encounter DCs through two routes: DCs residing in the lamina propria sample luminal bacterial antigens by passing their dendrites between IECs into the gut lumen, and DCs can also interact directly with bacteria that have gained access to the dome region of the gut-associated lymphoid tissue (GALT) through specialized epithelial cells, termed microfold or M cells. The interaction of the host cells with microorganism-associated molecular patterns (MAMPs) that are present on the surface macromolecules of probiotic bacteria will induce a certain molecular response. The host pattern recognition receptors (PRRs) that can perceive probiotic signals include Toll-like receptors (TLRs) and the C type lectin DC-specific intercellular adhesion molecule 3-grabbing non-integrin (DC-SIGN). Some molecular responses of IECs depend on the subtype of cell, for example, Paneth cells produce defensins and goblet cells produce mucus. Important responses of DCs against probiotics include the production of cytokines, major histocompatibility complex molecules for antigen presentation, and co-stimulatory molecules that polarize T cells into T helper or CD4+CD25+ regulatory T cells in the mesenteric lymph nodes (MLNs) or subepithelial dome of the GALT.
Host interactions of probiotic bacterial surface molecules: comparison with commensals and pathogens.



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