FOR MEDICAL COMMUNITY

KyberPlus: Mucous membrane diagnostic

if suspected

Several hundred square meters of intestinal mucous membrane run through the body. If the mucous membrane is inflamed or more permeable to allergens, pathogens and other noxious substances, this has serious effects on health. The body's own protection against infection on the intestinal mucosa can also be reduced.

The KyberPlus modular system, made up of various biochemical parameters, detects such disorders on the intestinal mucosa and thus reveals the causes of unclear abdominal complaints.

The KyberPlus diagnostics are suitable for clarifying:

    •     Maldigestion
    •     Pancreatic insufficiency
    •     Chronic pancreatitis
    •     Reduced mucosal protection
    •     Acute and chronic intestinal inflammation
    •     Relapse predictions in Crohn's disease
    •     Necrotizing enterocolitis
    •     Enteral protein loss syndrome
    •     Permeability disorders of the intestinal mucosa
    •     Gluten sensitive enteropathy
    •     Intestinal parasitosis
    •     Differentiation between food allergy and food intolerance
    •     Proof of effectiveness of an elimination diet     

      KyberPlus parameter

      Clarify digestive disorders

      If the digestive performance is impaired, nutrient deficiencies can occur. The cause can be a pancreatic insufficiency or a lack of bile acids. If intestinal bacteria break down proteins to a greater extent, hepatotoxic substances develop.

       

       

      Digestion residues

      The quantitative detection of digestive residues in the stool is useful for unclear gastrointestinal complaints. As a rule, there are only small amounts of undigested food residues in the stool; the daily fat and nitrogen excretion is relatively constant in a healthy person.

      If the excretion of fat or nitrogen increases to pathologically high concentrations, a digestive disorder such as maldigestion may be present. If it persists for a long time, maldigestion can turn into malabsorption - with the associated lack of vitamins, minerals and trace elements.

      Causes of maldigestion are:

      •     Exocrine pancreatic insufficiency or
      •     Bile acid deficiency

      In exocrine pancreatic insufficiency, there is a deficiency in the lipase, trypsin and chymotrypsin enzymes that break down fat and protein, and the corresponding food components are no longer sufficiently broken down. High molecular fats and proteins are no longer absorbed and accumulate in the stool.

      Bile acid deficiency can occur when the intestinal flora breaks down conjugated bile acids to a greater extent. This mainly happens when the small intestine is overgrowed with bacteria from the colon flora - the so-called overgrowth syndrome. In ileum dysfunction, bile acid reabsorption in the terminal ileum can be disturbed. In both cases there are too few bile acids to sufficiently emulsify the dietary fats. This increases the fat concentration in the stool.

      The water content of the stool indicates whether the patient is suffering from constipation or diarrhea, and provides an indication of how much fiber the patient is consuming.

      Stool samples for the detection of digestive residues must not exceed the maximum sample transport time of 4 days.

      Reference range: fat: increased from> 3.5%, nitrogen: increased from> 1.0%, water: increased from> 80.0%

      Iso-fatty acids

      When protein is broken down, the intestinal bacteria form iso-fatty acids: i-valeric acid and i-butyric acid. In parallel to the iso-fatty acids, hepatotoxic metabolites such as indole, skatole and phenol are formed. The iso-fatty acids therefore serve as an indirect marker for substances that are harmful to the liver.

      Raised iso-fatty acid levels can be caused by impaired digestive performance, high cell counts of proteolytic bacteria and a diet rich in animal proteins.

       

       

      Bile acids

      The determination of the bile acid concentration in the stool is carried out if a bile acid loss syndrome is suspected.

      The body produces around 700 ml of bile a day, approximately 12 percent of which consists of bile acids or bile salts. The bile acids are normally subject to the enterohepatic cycle. This means that most of the bile acids secreted into the duodenum are reabsorbed in the terminal ileum and returned to the liver. About 0.6 g of bile acids are lost in the stool every day and have to be synthesized again.
      An ileum dysfunction means that the bile acids are insufficiently absorbed. The body excretes them increasingly through the stool.

      Common causes of bile acid loss are:

      • Ileitis in Crohn's disease
      • Resection of the terminal ileum
      • Bacterial small bowel overgroth (SBOG)

      Bacterial decomposition of bile acids in an intestinal blind sac rarely leads to an indirect loss of bile acid.

      Compensated bile acid loss syndrome: If the body can replace the lost bile acids in sufficient quantities, fat digestion still works. Large amounts of bile acids can be detected in the stool, the fats are in the normal range.

      Decompensated bile acid loss syndrome: The liver is no longer able to synthesize enough new bile acids. The disease results in a functional bile acid deficiency and thus in impaired fat digestion.

      The bile acids are osmotically effective in the colon, which is why chologenen diarrhea occurs. If a functional bile acid deficiency has already occurred, the impaired fat absorption triggers steatorrhea.

      The reduced fat absorption also causes a lack of fat-soluble vitamins (hypovitaminosis) and weight loss. As the anus becomes irritated, anal eczema occurs. When the emulsifying bile acids are absent, the bile has increased lithogenicity. Gallstones are the result.

      Stool samples for the detection of bile acids must not exceed the maximum sample transport time of 3 days.

      Reference values (bile acids/100ml stool):

      •      <66 μmol                    reduced
      •     66 - 715 μmol              normal
      •     715.1 - 900 μmol        slightly increased
      •     900.1 - 1200 μmol     significantly increased
      •     > 1200 μmol                highly increased

      Pancreas Specific Elastase 1

      A reduced concentration of pancreatic-specific elastase 1 in the stool indicates chronic pancreatitis or pancreatic insufficiency. As described above, pancreatic insufficiency leads to maldigestion. For this reason, patients at risk - for example with diabetes or gallstones - are recommended to have their pancreas-specific fish elastase 1 determined as a preventive examination. Because of the reduced calcium absorption, the examination is also indicated in patients at risk of osteoporosis.

      The enzyme pancreas-specific elastase 1 is produced in the pancreas and excreted into the duodenum via the papilla vateri. Since the enzyme is not broken down, it can be detected in the stool. The pancreas-specific elastase 1 is a proteolytic glycoprotein with a molecular weight of about 28 kilodaltons (kDa).

      Stool samples for the detection of pancreas-specific elastase 1 must not exceed the maximum sample transport time of 7 days.

      Reference range for pancreas-specific elastase 1 (E1):

      •     ≥ 200 μg E1 / g stool: normal exocrine pancreatic function
      •     100 to 200 μg E1 / g stool: mild to moderate exocrine pancreatic insufficiency
      •     <100 μg E1 / g stool: severe exocrine pancreatic insufficiency

      Biomarkers of the mucosal defence

      The biomarkers sIgA, EPX and β-Defensin 2 demonstrate the ability of the intestinal mucosa to repel pathogens, antigens and pollutants.

       

       

      Secretory IgA

      The production of secretory immunoglobulin A (sIgA) is reduced in the case of recurrent infections of the mucous membranes, atopy and humoral immunodeficiencies. Adequate sIgA production is important for effective mucosal  protection. Every day  a person secretes between 5 and 15 g of sIgA on the mucous membranes: tears, breast milk, saliva, the mucus of the bronchi, the urogenital and gastrointestinal tracts contain the immunoglobulin.

      SIgA binds to bacteria or viruses that have penetrated the gastrointestinal tract or bronchi; then the peristalsis of the intestine and the ciliated epithelium of the bronchal tubesi transport the sIgA along with bound pathogens.

      The secretory immunoglobulin A consists of two IgA molecules, the J chain and a secretory component. The polymerisation to sIgA only takes place in the mucous membranes; it is necessary for the transport through the epithelial cells. The IgA molecules bind to a receptor on the side facing away from the lumen and are channeled through the cell. When released in the lumen, part of the receptor remains attached to the IgA dimer - the secretory component. It protects the immunoglobulins from being broken down by digestive enzymes and from microbial attack.

      Stool samples for the detection of secretory immunoglobulin A must not exceed the maximum sample transport time of 2 days.

      Reference range: increased from> 2040 μg / ml sIgA in the faeces

      β-defensin 2

      The human skin and the mucous membranes form substances with an antibiotic effect - the defensins. They are part of the chemical barrier that, together with the physical barrier, protects  the epithelial cells from intruders. Defensin production disorders play a role in allergies such as neurodermatitis or bronchial asthma and in inflammatory bowel diseases. The β-defensins are the most common of all defensin types. The skin and mucous membrane form β-defensin 2 when they come into contact with bacteria or when an inflammatory reaction starts. Preparations containing bacteria can stimulate defensin synthesis in the intestine and thus strengthen the mucous membrane barrier.

      Stool samples for the detection of β-defensin 2 must not exceed the maximum sample transport time of 3 days.

      Reference range: increased from> 60 ng / ml


      Permeability of the mucosal epithelium (leaky gut)

      Zonulin and α1-antitrypsin indicate the permeability of the intestinal epithelium. If the values ​​of the biomarkers in the stool are increased, the risk of inflammation of the intestinal epithelium, allergies and metabolic endotoxinemia increases.

      Zonulin

      The regulator protein Zonulin is a suitable marker for measuring the permeability of the intestinal mucosa. Zonulin regulates the exchange of fluid, macromolecules and leukocytes between the bloodstream and the intestinal lumen. It also protects the subepithelial layers.

      Various stimuli cause the intestinal epithelial cells to release zonulin into the intestinal lumen and blood vessels. Examples are direct contact with bacteria when the intestinal mucus layer is missing or interrupted and contact with gliadin. The zonulin binds to receptors on the surface of the intestinal epithelial cells through which the cell's cytoskeleton contracts. As a result, the tight junctions open. If the zonulin-mediated opening of the tight junctions takes place repeatedly and intensely, the leaky gut syndrome develops.

      Stool samples for the detection of zonulin must not exceed the maximum sample transport time of 4 days.

      Reference range: borderlinel from> 78.0 ng / ml stool or> 48.0 ng / ml serum

      α1-antitrypsin

      α1-Antitrypsin (α1-AT) serves as a marker for inflammation and permeability disorders in:

      •     Inflammatory diseases of the gastrointestinal tract
      •     Crohn's disease (prognosis of a relapse)
      •     Necrotizing enterocolitis
      •     Suspected enteral protein loss syndrome
      •     Permeability disorders of the intestinal mucosa
      •     Gluten sensitive enteropathy.

      The liver in particular produces the protein α1-antitrypsin. It accounts for up to 90 percent of all α1 globulins. α1-Antitrypsin is a protease inhibitor, i.e. it inhibits proteolytic enzymes and thus prevents the breakdown of blood protein. Α1-antitrypsin inhibits the enzyme elastase particularly strongly, but with decreasing effectiveness also the enzymes trypsin, plasmin, thrombin and plasminogen. As a protease inhibitor, α1-antitrypsin itself is only slightly broken down and is therefore well suited as a marker. When there is inflammation, the body produces more α1-antitrypsin.

      Stool samples for the detection of α1-antitrypsin must not exceed the maximum sample transport time of 3 days.

      Reference range: increased from 56 mg α1-AT / dl stool.


      Markers of inflammation

      Calprotectin, lysozyme and lactoferrin reflect the extent of the granulocyte immigration into the intestine and thus indicate inflammation of the intestinal mucosa. The combination of the markers increases the sensitivity.

      The advantage of the fecal inflammation markers: the collection of samples is simple and non-invasive and the determination is inexpensive.

      Calprotectin

      Calprotectin is suitable for diagnosing and monitoring the course of chronic inflammatory bowel diseases, Crohn's disease and ulcerative colitis. It is also used for the diagnosis of exclusion in irritable bowel syndrome.

      The neutrophil granulocytes and the monocytes form the fecal calprotectin as a calcium-binding protein complex. Calptrotectin is a sensitive marker for inflammatory bowel disease.  In one study, calprotectin showed a sensitivity of 82% and a specificity of 87% in distinguishing between irritable bowel syndrome and inflammatory bowel disease.

      Areas of application:

      • Acute inflammatory processes in the delimitation of functional complaints     
      • Activity monitoring of already known chronic inflammatory diseases such as Crohn's disease and ulcerative colitis

      Stool samples for the detection of calprotectin must not exceed the maximum sample transport time of 4 days.

      Reference range: increased from 50 μg calprotectin / g stool

      Lysozyme

      The lysozyme levels in the stool are an indicator of the extent to which leukocytes have migrated into the intestinal lumen. Ulcerative colitis and Crohn's disease patients have high levels of lysozyme in their stool.

      Lysozyme can be found in neutrophils, macrophages and Paneth cells and is found in saliva, sweat, nasal secretions and tear fluid. The enzyme breaks down sugar chains in the bacterial cell wall and thus initiates lysis of the bacterial cell. However, the enzyme only attacks gram-positive bacteria such as streptococci and staphylococci, because in gram-negative bacteria the outer membrane protects the sugar chains from degradation.

      In addition, lysozyme has an indirect bactericidal effect, as it increases the activity of immune antibodies. Dissolving the agglutination of microorganisms improves uptake and destruction by macrophages.

      Stool samples for the detection of the lysozyme must not exceed the maximum sample transport time of 2 days.

      Reference range: increased from 600 ng / ml lysozyme in the stool.

      Lactoferrin

      The activity of the disease in patients with chronic inflammatory bowel disease (IBD) can be assessed via the lactoferrin concentration in the stool. The parameter is also suitable for monitoring the success of therapy in IBD.

      Lactoferrin is an iron-binding protein in the secondary granules of the neutrophil granulocytes. It is part of the innate immune defense on the mucous membranes. When there is inflammation, the neutrophils release lactoferrin to kill pathogens. By binding iron, lactoferrin has an antimicrobial effect.

      An inflammation in the intestines causes the lactoferrin levels in the stool to rise. In non-inflammatory diseases such as irritable bowel syndrome, on the other hand, the lactoferrin values ​​are in the normal range.

      Application areas:

      •     Acute inflammatory processes in the delimitation of functional complaints
      •     Acute bacterial infections of the colon
      •     Activity monitoring of already known, chronic inflammatory diseases such as Crohn's disease
            and ulcerative colitis
      •     Tumor search in combination with the M2-PK tumor marker

      Stool samples for the detection of lactoferrin must not exceed the maximum sample transport time of 3 days.

      Reference range: increased from 7.24 μg per g stool.

      Eosinophil protein X

      The amount of eosinophil protein X (EPX) circulating is a reflection of the body's inflammatory status. EPX is suitable for

      • the detection of acute or chronic intestinal inflammation
      • to differentiate between food allergy and food intolerance
      • to test the effectiveness of an elimination diet
      • for the detection of intestinal parasitosis.

      Eosinophils belong to the group of leukocytes. They occur more frequently in foci of inflammation and in response to parasite infections. In the cytoplasm of the eosinophils there are granules with positively charged proteins. The granule proteins are basic and bind to strongly acidic dyes. The special affinity for the red-orange pigment eosin gave the eosinophils their name. During degranulation, for example, the eosinophils release EPX into the surrounding tissue. Granule proteins like EPX can kill parasites, but they can also cause tissue damage associated with inflammatory diseases.

      Activation of the eosinophils can be observed in many inflammatory processes. Examples are bronchial asthma, atopic dermatitis, rhinitis, allergic eye infections, allergic middle ear effusions, parasite and bacterial infections, autoimmune diseases and the chronic fatigue syndrome.

      Stool samples for the detection of EPX must not exceed the maximum sample transport time of 7 days.

      Reference range: increased from 1700 ng EPX / ml stool


      Mucosal supply and bacterially mediated saturation

      The microbiota forms the short-chain fatty acids acetic acid, propionic acid and butyric acid. By them it nourishes the intestinal mucous membrane and influences our feeling of satiety.

       

       

      Short chain fatty acids

      The intestinal microbiota forms acetic acid, propionic acid and butyric acid and intervenes in our metabolism via the short-chain fatty acids. The acetic acid formed by bacteria increases the feeling of hunger and stimulates gluconeogenesis and liponeogenesis. This provides the organism with additional calories. Propionic acid works in the opposite direction: it increases the feeling of satiety, lowers the cholesterol level and improves insulin sensitivity. Butyric acid is particularly important for the intestinal epithelium: It provides 80 percent of the nutrition for the epithelial cells and thus ensures the functionality of the intestinal mucosa. A reduced butyric acid content indicates an insufficient supply of the intestinal epithelium.

      Stool samples for the detection of short-chain fatty acids must not exceed the maximum sample transport time of 2 days.

       

       


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