Short-time high-temperature processing for microbial preparations with anti-inflammatory properties
FIELD: medicine, pharmaceutics.
SUBSTANCE: present invention refers to microbiology, namely to using bacteria, and describes a composition containing microorganisms, a method for preparing probiotics with the anti-inflammatory action and a method for preparing an anti-inflammatory composition containing the probiotics. The composition according to the invention is characterised by the microorganism count subject to the short-time high-temperature processing at 120-140°C for 5-15 seconds.
EFFECT: invention can be used for preparing the compositions for treating or preventing the inflammatory disorders for the probiotics for dairy products.
10 cl, 7 dwg
The present invention mainly relates to microorganisms. In particular, the present invention relates to microorganisms subjected to high temperatures, such as probiotics and starter cultures for dairy products, as well as applications of these bacteria. One of the embodiments of the present invention relates to microorganisms subjected to high temperatures, such as probiotics and starter cultures for dairy products, and their use in obtaining compositions for the treatment or prevention of inflammatory disorders.
Probiotics often give the following definition: "live microorganisms, which when introduced into the body-the owner of the right quantities of benefit to health (regulations Control quality control of food, drugs and cosmetics USA and world health organization). Accordingly, most of the publications deal with living probiotics. However, in a number of studies have investigated the beneficial effects on health, provided nerealizirane bacteria, and most of them were obtained indicate that inactivation of probiotics, for example by heat treatment, leads to the loss of desired health benefits (Rachmilewitz, D., et al., 2004, Gastroenterology 126:520-528; Castagliuolo et al., 2005, FEMS Immunol.Med.Microiol. 43:197-204; Gill, H. S. and K. J. Rutherford, 2001,Br.J.Nutr. 86:285-289; Kaila, M., et al., 1995, Arch.Dis.Child 72:51-53.).
However, some studies have shown that inactivated probiotics can to some extent to maintain a positive influence on health (Rachmilewitz, D., et al., 2004, Gastroenterology 126:520-528; Gill, H. S. and K. J. Rutherfurd, 2001, Br.J.Nutr. 86:285-289).
About the use of probiotics for the treatment and prevention of inflammatory diseases of the gastrointestinal tract was reported in the press, and recently published review on this topic (Dotan, I. and D. Rachmilewitz. 2005; Curr.Opin.Gastroenterol, 21:426-430). For example, it is shown that a highly concentrated mixture of eight live bacteria-probiotics (VSL#3) are effective for prophylaxis (Gionchetti, P., et al., 2003, Gastroenterology 124:1202-1209) and treatment (Gionchetti, P., et al., 2000, Gastroenterology 119:305-309; Mimura, T., et al., 2004, Gut 53:p.108-114) recurrent or persistent inflammation of the small bowel pockets (paucity) after surgery in humans. Notable work Rachmilewitz, D., et al., (2004, Gastroenterology 126:520-528), which was used as the animal model, namely mice with ulcerative colitis, induced doctranslate sodium (DSS); these authors reported that the introduction of live or exposed to gamma radiation VSL#3 was protecting animals from colitis, a VSL#3, inactivated by heat treatment, the protective effect did not possess. Similarly subjected to heat treatment L. crispatus not Conn is whether from DSS-induced colitis, while alive, these bacteria was clearly reduced the loss of body weight and the activity of myeloperoxidase (MPO) in the intestine (Castagliuolo et al., 2005, FEMS Immunol.Med.Environ. 43:197-204). These studies suggest that in relation to inflammatory processes in the gut probiotics live better than in saralicious.
Found that inactivated bacteria L. reuteri (killed by heat treatment or exposed to gamma radiation) is not able to reduce induced tumor necrosis factor (TNF-α) formation of interleukin-8 (IL-8) cells T, while alive, these bacteria have a marked beneficial effect (MA, D., et A1., 2004, Infect.Immun. 72:5308-5314).
It is clear that inactivated probiotics significantly easier to use in practice, for example in the food industry, and/or store. Therefore, a very promising opening that consumption of inactivated probiotics can have a positive effect on the body, although this effect is generally weaker than that of live bacteria, or nonexistent.
Reported receiving rereplacenocase strains of bacteria-probiotics through action on the corresponding live bacteria high temperature, gamma radiation, ultraviolet radiation or chemical agents (formalin, paraformaldehyde). But l will find work, which would equal the dependence of the biological functions obtained rereplacenocase bacteria from the applied method of inactivation. In addition to the above-mentioned studies, in one study it was reported that bacteria exposed to gamma radiation, and bacteria that have undergone heat treatment, induced different levels of secretion of cytokines by epithelial cells in vitro (Wong, S. and Z. Ustunol. 2006, J. Food Prot, 69:2285-2288).
However tried out in practice and described in the literature, the methods of transforming probiotic strains in rereplacenocase, as a rule, not applicable or hardly applicable in an industrial environment, especially in the food industry.
The method of manufacture of the majority of commercially available products containing probiotics, includes a heat treatment. It would be so efficiently conduct heat treatment of probiotics together with izgotovlivaem product or at least a similar manner so that probiotics have maintained or improved their positive properties or even bought new and useful for consuming organism properties.
But described in the literature, the methods of heat treatment used to inactivate bacteria, assume, as a rule, lasting from 20 minutes to 1 hour or more) temperatures from 40 to 100°C, which is not easy to implement in p is myshlennyh scale. In addition, this heat treatment resulted, as a rule, to the loss of probiotic activity, at least partially.
On an industrial scale is applied for a short heat treatment, for example ultra pasteurization (aseptic pasteurization). This heat treatment reduces bacterial load in the product and the time of its manufacture, thereby reducing the degree of damage of nutrients.
Therefore, it would be highly desirable to have products of microbial cells obtained from viable microorganisms, such as bacteria-probiotic or starter cultures for dairy products, which could be heat treated in the same way as manufactured products, which was acquired or improved healthy consumer properties, for example, contributing to the prevention or treatment of inflammatory disorders.
Thus, one of the objectives of the present invention is the provision of a composition containing microorganisms, such as bacteria-probiotics or starter cultures for dairy products, which can be subjected to heat treatment in an industrial environment to reduce the number of viable bacterial cells and that as a result of this heat treatment acquire or improve useful for ZV is cited consumer properties.
The authors of the present invention unexpectedly found that this problem can be solved by the object of the invention according to the independent claims. Further development of the present invention receives dependent claims.
The authors of the present invention for the first time demonstrated that microorganisms, such as probiotic strains of bacteria or starter cultures for dairy products, past short-time heat treatment at high temperature, anti-inflammatory immunogenic properties without regard to their original properties. In particular, as a result of such heat treatment may develop new anti-inflammatory profile or improving existing ones.
Thus, it is possible to get rereplacenocase microorganisms that have anti-inflammatory immunogenic properties, by applying certain parameters of heat treatment, the respective normally used in an industrial environment regimes of heat treatment, even if the original living microorganisms were characterized as anti-inflammatory strains.
This effect was observed, for example, a number of microorganisms, namely probiotic representatives of lactobacilli and bifidobacteria, as well as starter cultures for Moloch the th product.
Rereplacenocase probiotic microorganisms have the advantage that they are easier to manipulate than the corresponding live strains. In addition, they are much better able to withstand storage and require less stringent conditions of the package.
Rereplacenocase probiotic microorganisms could develop a variety of products, which by their nature do not allow the addition of live probiotics without additional measures for their protection. This is important, for example, in the production of cereal bars, fruit juices, UHT beverages, beverages with a long shelf life and other
In addition, if the consumer has a weakened immune system, live probiotics can be applied only in exceptional cases because of the risk of bacteremia. In this paper the authors propose a method of obtaining viable bacteria with anti-inflammatory properties regardless of their source immunogenic profile.
Also rereplacenocase probiotic microorganisms allow dissolution in hot form, for example, powdered compositions, with lasting benefits for the health of the consumer properties.
To the authors ' knowledge of the invention, not previously reported short-term high-temperature processing on the I get rereplacenocase bacteria with anti-inflammatory properties.
Accordingly, in one of the embodiments of the present invention features a composition containing microorganisms subjected to short-term exposure to high temperature is at least 71,5°C for at least 1 second.
In the present invention are preferably used food microorganisms. The microorganism is called food, if officially admitted its consumption by humans or animals.
In one of the embodiments of the present invention, the microorganisms can be probiotics.
In the context of the present invention probiotics are referred to as "the products of microbial cells or components of microbial cells that are beneficial for the health or life of the host body" (Salminen S, Ouwehand A. Benno Y. et al "Probiotics: how should they be defined" Trends Food Sci. Technol. 1999:10107-10).
In another embodiment of this invention, the microorganisms can be starter cultures for dairy products.
In this invention the microorganisms are subjected to a short heat treatment, preferably at a temperature of from about 71.5 to about 150°C for a time from about 1 to about 120 seconds.
The preferred heat treatment of microorganisms at a temperature of from about 90 to about 140°C., for example in the range of 90-120°C. for a time from about 1 to about 30 seconds.In one embodiment of the present invention, such high-temperature processing makes the microorganisms in at least partially rereplacenocase.
The term "rereplacenocase the microorganisms include microorganisms, such as bacteria-probiotics or starter cultures for dairy products, which have undergone a heat treatment. This includes inactivated, dead, non-viable and/or fragments (e.g., DNA, metabolites, cytoplasmic connections and/or material of the cell walls of microorganisms.
The term "rereplacenocase" means that viable cells and/or colony-forming units cannot be detected by commonly used methods of planting/cultivation. Such methods are described in the guide: James Monroe Jay, Martin J. Loessner, David A. Golden. 2005. Modem food microbiology. 7th edition, Springer Science, New York, N. Y. 790 p. As a rule, the absence of viable cells is demonstrated as follows: no visible eye colonies on agar or increase turbidity in the liquid culture medium after sowing preparations of bacterial cells in various concentrations (rereplacenocase samples) and incubation under suitable conditions (aerobic and/or anaerobic for at least 24 hours).
High temperature processing can be performed at normal atmospheric pressure, h is also at high pressure. Typically, the pressure is in the range from 1 to 50 bar, preferably from 1 to 10 bar, more preferably from 2 to 5 bar. Such pressure preferably, if probiotics are subjected to heat treatment in an environment that upon heating of the liquid or solid. What pressure would be the best, depends on the nature of the composition, which must contain these microorganisms, and from the accompanying temperature.
Heat treatment according to this invention can be carried out in the temperature range of about 71.5-150°C, preferably about 90-120°C., even more preferably of about 120-140°C.
Heat treatment according to this invention can be accomplished in a short period of time duration of about 1-120 seconds, preferably about 1-30 seconds, even more preferably about 5-15 seconds.
These time limits apply to the time during which the microorganisms, such as probiotics and starter cultures for dairy products, are exposed to this temperature. Note that the time of heat exposure may vary depending on the nature and quantity of the composition, which must contain probiotics, and designs used heating equipment.
Usually, however, the composition of this invention and/or the microorganisms are exposed to kratkovremennoi high-temperature treatment (HTST), instant pasteurization or ultra-pasteurization (UHT).
Processing, abbreviated UHT (ultra heat treatment), involves brief exposure to very high temperature, which is achieved at least partial sterilization; its duration is about 1-10 seconds, during which the temperature exceeds 135°C (275°F), which is required for the destruction of bacterial spores in milk. For example, in milk subjected to temperatures above 135°C, the bacterial load required for processing time (2-5 seconds) is reduced to the extent that it makes possible the operation in a continuous stream.
System for ultra pasteurization, there are two basic types - direct and indirect. In direct systems, the product is treated by injection or steam injection and indirect systems, the product is heated using a heat exchanger (plate, tubular or scraper). System combination of ultra-pasteurization can be applied at any stage or several stages of the manufacturing process of the product.
Short-term high-temperature processing (HTST) is defined as follows: this method of pasteurization, designed to reduce the number of viable microorganisms in milk 5 orders of magnitude, i.e., killing 99,9999%. This treatment is considered sufficient, the La destruction of nearly all yeast and mold cells and normal bacteria causing spoilage of dairy products, as well as ensuring the destruction of the normal pathogenic organisms resistant to heat. During HTST treatment, the milk is heated to 71,7°C for 15-20 seconds.
Instant pasteurization is a method of thermal pasteurization of perishable beverages such as vegetable and fruit juices, beer and dairy products. It is performed before filling the containers for the product to destroy causing spoilage microorganisms, improve safety and extend shelf life. For this liquid in a continuous controlled flow is subjected to temperatures from 71,5°C to 74°C for 15-30 seconds.
In the context of the present invention, the term "short-term high-temperature processing" includes, for example, short-term high-temperature pasteurization (HTST, UHT) and instant pasteurization.
Having lost the ability to replicate in the short-term high-temperature processing obtained rereplacenocase probiotics are improved or new useful properties, as described above.
Now any number rereplacenocase microorganisms discussed above. But, as a rule, it is considered desirable that nerealizirane became at least 90% of probiotic organisms, preferably m is Nisha least 95%, more preferably at least 98%, most preferably at least 99%, better yet at least about 99.9%, ideally all probiotics become nerealizirane.
In one of the embodiments of the present invention, all microorganisms are nerealizirane.
In the framework of the present invention can be used all microorganisms. Preferably used microorganisms were acceptable in food products. Typical microorganisms that are acceptable in food products, is probiotics.
Preferably in the present invention are used probiotics selected from the group consisting of the representatives of the genera Bifidobacterium, Lactobacillus, Lactococcus, Streptococcus, Candida, or mixtures thereof.
For example, probiotics according to the present invention can be selected from the group consisting of Bifidobacterium longum, Bifidobacterium lactis, Bifidobacterium animalis, Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium adolescentis, Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus salivarius, Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus johnsonii, Lactobacillus plantarum, Lactobacillus fermentum, Lactococcus lactis, and/or mixtures thereof.
Starter cultures for dairy products according to the present invention can be selected from the group consisting of Propionibacterium, Streptococcus thermophilus, Lactococcus lactis, Lactococcus diacetylactis, Lactococcus cremoris, Lactobacillus bulgaricus, Lactobacillus helveticus, Lactobacillus delbrueckii, and mixtures thereof.
It is essential that probiotics are subjected to the SJ high-temperature processing, for example to temperatures for 71.5-150°C, for short periods of time (1-120 seconds). This treatment can be a viable and/or non-viable probiotics.
You can perform repeated high-temperature processing, such as the impact of temperature for 71.5-150°C for short time (1-120 seconds).
In one embodiment of the present invention, probiotici selected from the group consisting of the genera Bifidobacterium, Lactobacillus and Escherichia or combinations thereof.
Typical examples of the Bifidobacterium is Bifidobacterium longum, Bifidobacterium breve or Bifidobacterium lactis. Typical examples of lactobacilli Lactobacillus paracasei, Lactobacillus casei, Lactobacillus acidophilus or Lactobacillus rhamnosus. A typical example of Escherichia strains of Escherichia coli with the desired useful properties.
In one embodiment of the present invention starter cultures for dairy products serve as Streptococcus thermophilus, Lactobacillus helveticus, Lactobacillus delbrueckii. Lactococcus lactis.
Specifically, probiotics can be, for example, from the group consisting of Bifidobacterium longum NCC 3001, Bifidobacterium longum NCC 2705, Bifidobacterium breve NCC 2950, Bifidobacterium lactis NCC 2818, Lactobacillus paracasei NCC 2461, Lactobacillus casei NCC 4006, Lactobacillus casei ACA-DC 6002 (NCC 1825), Lactobacillus rhamnosus NCC 4007, Lactobacillus acidophilus NCC 3009, Escherichia coli Nissle 1917 or combinations thereof.
Also starter cultures for dairy products you can choose, for example, from the group consisting of Streptococcus thermophilus NCC 2019, Streptococcus thermophilus NCC 2059, Lactobacillus delbrueckii subsp.bulgaricus NCC 15 Lactococcus lactis NCC 2287.
All strains according to the invention deposited according to the Budapest Treaty under the following registration numbers:
|Bifldobacterium longum NCC 3001:||ATCC BAA-999|
|Bifidobacterium longum NCC 2705:||CNCM I-2618|
|Bifldobacterium breve NCC 2950||CNCM I-3865|
|Bifidobacterium lactis NCC 2818:||CNCM I-3446|
|Lactobacillus paracasei NCC 2461:||CNCM I-2116|
|Lactobacillus rhamnosus NCC 4007:||CGMCC 1.3724|
|Streptococcus themophilus NCC 2019:||CNCM I-1422|
|Streptococcus themophilus NCC 2059:||CNCM I-4153|
|Lactococcus lactis NCC 2287:||CNCM I-4154|
|Lactobacillus casei NCC 4006:||CNCM I-1518|
|Lactobacillus casei NCC 1825:||ACA-DC 6002|
|Lactobacillus acidophilus NCC 3009:||ATCC 700396|
|Lactobacillus bulgaricus NCC 15:||CNCM 1-1198|
|Escherichia coli Nissle 1917:||DSM6601.|
Compositions according to the present invention include microorganisms subjected to short-term exposure to high temperature, sufficient for at least partial treatment of inflammatory disorders and/or their complications. The number of microorganisms that are necessary and sufficient for the implementation of this task, called a "therapeutically effective dose". The number of microorganisms that are effective to achieve such a goal depends on a number of factors known to experts in the art, for example from the severity of the disease, patient's body weight and General state of health, as well as the nature of the food product, which are microorganisms.
When used in preventive composition according to the invention is introduced into the body of the consumer exposed to inflammatory disorder or otherwise experiencing the risk of such diseases in a quantity sufficient to at least partially reduce the risk of inflammatory disorders. This number of microorganisms called a "prophylactically effective dose." Again, the specific number depends on several individual factors, for example, from General health of the patient, his or her body weight, but also on the nature of the food product is one which are the microorganisms.
Specialists in the art will have no difficulty to find the right therapeutically effective dose and/or prophylactically effective dose.
Typically, the composition according to the invention contains microorganisms subjected to short-term exposure to high temperature, in a therapeutically effective dose and/or prophylactically effective dose.
In a typical case, therapeutically effective dose and/or prophylactically effective dose of 0.005 mg/day to 1000 mg/day rereplacenocase microorganisms subjected to short-term exposure to high temperature.
In specific terms rereplacenocase microorganisms subjected to short-term exposure to high temperature, may be present in compositions according to the invention in an amount corresponding to 104-1012equivalents CFU per 1 g of the composition in the dry state. It is clear that rereplacenocase microorganisms do not form colonies, so that this definition should be understood as the number rereplacenocase microorganisms obtained from 104-1012CFU/g bacteria can replicate. This includes microorganisms are inactivated, non-viable or dead or present in the form of fragments (DNA, cell wall, the component of the s cytoplasm). In other words, the number of microorganisms contained in the compositions of this invention, expressed as colony-forming ability (CFU) of such number of microorganisms, as if all these microorganisms were alive without regard to whether they actually nerealizirane (inactivated or dead, fragmented, or presents a mixture of any or all of these States).
Preferably the microorganisms according to the invention are present in the composition in an amount equivalent to from 104up to 109CFU/g of the composition in the dry state, more preferably in an amount equivalent to from 105up to 108CFU/g of the composition in the dry state.
The composition according to the invention may be in any form. It can be intended for oral or other enteral administration, for parenteral administration (subcutaneous or intramuscular), intravaginal, intrarectal, local, or intraocular injection. This can be, for example, the pharmaceutical composition, nutraceuticals, dietary Supplement, cosmetic composition, animal feed, food product or beverage.
Food products according to the invention include dairy products, for example products of lactic acid fermentation (yogurt, buttermilk, and other); ice cream; the oxygen is irovannoe milk; milk; cream; flavoured milk drinks; beverages based on milk whey; apical toppings; cream substitutes for coffee; chocolate; cheese products; soups; sauces; puree; dressings; puddings; creams; baby food; nutrient mixture (for example, for a complete supply: for infants, for children over one year of age, adolescents, adults, elderly or sick people; cereals and cereal bars (granola).
Beverages according to the invention include, for example, milk or yogurt drinks, milk drinks, protein drinks, coffee, tea, energy drinks, soy drinks, fruit and/or vegetable drinks, fruit and/or vegetable juices.
Cosmetic compositions according to the invention may include, for example, lotions, shampoos, eye drops, creams.
Biologically active additives or medicines may be, for example, in the form of tablets, capsules, lozenges, sachets, gels or liquids (for example, nutrient solutions).
The compositions of this invention may also contain protective hydrocolloids (such as gums, proteins, modified starches), binders, film forming agents, encapsulating agents/materials, materials in the cell walls, the components of the intercellular matrix, coatings, emulsifiers, over OSTO-active substances, solubilizing agents (oils, fats, waxes, lecithins, and other), absorbents, carriers, fillers, co-compounds, dispersing agents, moisturizing agents, technologically auxiliary substances (solvents), flowing agents, kusamakura agents, weighting agents, gelling agents, gel forming agents, antioxidants and antimicrobial agents. They may also contain conventional pharmaceutical additives and adjuvants, excipients and diluents, including, but not limited to, water, gelatin of any origin, vegetable gums, ligninsulfonate, talc, sugars, starch, Arabic gum, vegetable oils, polyalkylene glycols, flavoring agents, preservatives, stabilizers, emulsifiers, buffering agents, lubricating agents, colouring agents, moisturizing agents, fillers, etc.
Compositions according to the invention can contain organic or inorganic carrier suitable for oral or other enteral administration, as well as vitamins, minerals and other nutrients required by the body in small quantities, in accordance with state statutes, such as the norms of consumption of nutrients generally accepted in the United States.
Compositions according to the invention may also contain one who e agents, depending on the purpose of varying composition. For example, to reduce the discomfort caused an inflammatory disorder, the composition can be added analgesic or antipyretic agents.
In order to stabilize the composition and its components can be added stabilizing agents.
To give the song a pleasant smell/taste can be added flavouring agents, and/or to give the song a distinctive or attractive color - dyes.
In the compositions of this invention can be added prebiotics. Prebiotics can support the growth of probiotics prior to when their turn in rereplacenocase. Prebiotics may also act synergistically with viable bacteria-probiotics, which are present in the composition and/or can be added in it.
The term "prebiotic" means nevereverever food components that promote the growth of healthy microorganisms and/or probiotics in the gut. They are not broken down in the stomach and/or upper intestine and is not absorbed in the gastrointestinal tract of the consumer, and subjected to fermentation under the action of intestinal microflora and/or probiotics. Prebiotics are described, for example, in the work of Glenn R. Gibson and Marcel B. Roberfroid, Dietary Modulation of the Human Colonic Microbiota: Introducing the Concept f Prebiotics, J. Nutr. 1995 125:1401-1412.
Selection of probiotics for use according to the invention nothing specific is not limited; prebiotics used according to the invention include all the nutrients that promote the growth of probiotics, or healthy bacteria in the gut. Preferably prebiotics in this invention are selected from the group consisting of oligosaccharides, including, if necessary, include fructose, galactose, mannose; dietary fiber, particularly soluble fiber, soy fiber; inulin, or a mixture thereof. The preferred prebiotics according to the invention are fructo-oligosaccharides (FOS), galactooligosaccharides (GOS), isomaltooligosaccharide (IMO), xylooligosaccharide (XOS), arabinosylcytosine (AXOS), mannanoligosaccharide (MOS), soybean oligosaccharides, glycosylceramide (GS), lactosucrose (LS), lactulose (LA), platinochloride (RAO), maltooligosaccharide, gums and/or their hydrolysates, pectins and/or their hydrolysates.
In the composition according to the invention can also add more probiotics. Extras can be added to any of probiotic microorganisms. Preferably probiotics for this purpose are selected from the group consisting of Bifidobacterium, Lactobacillus, Lactococcus, Enterococcus, Streptococcus, Kluyveromyces, Saccharoymces, Candida, Escherichia, in particular selected from the group consisting of Bifidobacterium longum, Bfidobacterium lactis, Bifidobacterium animalis, Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium adolescentis, Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus paracasei, Lactobacillus salivarius, Lactobacillus lactis, Lactobacillus reuteri, Lactobacillus rhamnosus, Lactobacillus johnsonii, Lactobacillus plantarum, Lactobacillus salivarius, Lactococcus lactis, Enterococcus faecium, Saccharomyces cerevisiae, Saccharomyces boulardii, Escherichia coli, or mixtures thereof; more preferably the probiotic is selected from the group consisting of Bifidobacterium longum NCC 3001 (ATCC BAA-999), Bifidobacterium longum NCC 2705 (CNCM 1-2618), Bifidobacterium longum NCC 490 (CNCM 1-2170), Bifidobacterium lactis NCC 2818 (CNCM 1-3446), Bifidobacterium breve NCC 2950 (CNCM 1-3865), Lactobacillus paracasei NCC 2461 (CNCM 1-2116), Lactobacillus johnsonii NCC 533 (CNCM 1-1225), Lactobacillus rhamnosus GG (ATCC53103), Lactobacillus rhamnosus NCC 4007 (CGMCC 1.3724), Enterococcus faecium SF65 (NCC 2768; NCIMB10415) and mixtures thereof.
Compositions according to the invention can be designed for any mammal, but preferably they are for people and Pets.
The composition according to the invention may also contain all necessary in the daily diet vitamins and minerals in significant power quantities. For certain vitamins and minerals have installed the minimum required amount. Examples of minerals, vitamins and other nutrients optionally included in compositions according to this invention include vitamin a, vitamin B1, vitamin b2, vitamin b6vitamin B12, vitamin E, vitamin K, vitamin C, vitamin D, Valiev the Yu acid (vitamin b 9), Inositol (vitamin B8), Niacin (nicotinic acid, or vitamin PP), Biotin (vitamin b7), Pantothenic acid (vitamin B5), choline (vitamin b4), calcium, phosphorus, iodine, iron, magnesium, copper, zinc, manganese, chloride, potassium, sodium, selenium, chromium, molybdenum, taurine and L-carnitine. Mineral matter usually included in the composition in the form of salts. The presence in the composition of those or other mineral substances and vitamins, and their number depends on for whom is the song.
The composition according to the invention may contain at least one source of protein, at least one source of carbohydrate and at least one source of lipids.
As a source of protein you can use any suitable dietary protein, such as animal proteins (such as milk proteins, meat proteins and egg whites); plant proteins (for example, soybean protein, wheat protein, proteins of rice and pea proteins); mixtures of free amino acids or combinations thereof.
Especially preferred milk proteins (such as casein and whey proteins and soy proteins. As for whey protein, the protein source can be sour or sweet whey or mixtures thereof and may include alpha-lactalbumin and beta-lactoglobulin in the desired ratio.
Preferably, especially in the case of mixtures for children who whom power to the source of protein was based on modified sweet whey. Sweet whey is a readily available by-product of the manufacture of cheese; it is often used in the production of mixtures based on cow's milk.
Proteins in the composition according to the invention can be intact or gidrolizovannykh or may be a mixture of intact and hydrolysed proteins. It is desirable to use a partially hydrolyzed protein (degree of hydrolysis 2-20%).
If you need hydrolysed proteins, the process of hydrolysis can be performed as desired, using known in the field methods. For example, you can get hydrolyzed whey protein by enzymatic hydrolysis of the corresponding fractions of whey in one or several stages.
If the composition according to the invention contains a source of protein, the amount of protein in it or protein equivalent is, as a rule, from 1.6 to 7.5 g/100 kcal of supplying energy to birds songs.
In particular, in the case of a nutritious source of protein mixtures should provide the minimum needs of the body for essential amino acids.
If the composition of this invention contains a source of carbohydrates, the nature of the used carbohydrates nothing specific is not limited. You can use any suitable carbohydrate such as sucrose, lactose, glucose, fructose, dried corn syrup, maltodextrins, starch and mixtures thereof. You can use a combination of different sources of carbohydrates. Preferably carbohydrates provide from 30 to 80% of supplying energy to birds songs. For example, the composition of this invention may contain a source of carbohydrate in the amount of 9 to 18 g/100 kcal of supplying energy to birds songs.
If the composition according to the invention contains a source of lipids, the nature of the used lipids nothing specific is not limited. If the composition of this invention contains a source of lipids, it can provide from 5 to 70% of supplying energy to birds songs. The composition can include long-chain n-3 and/or n-6 polyunsaturated fatty acids, such as DHA, ARA and/or EPA. The desired composition of the fat can be achieved by using a mixture of improved rapeseed oil, corn oil, sunflower oil with high oleic acid content, as well as triglycerides of medium chain length. The composition according to this invention may contain a source of lipids in the amount of 1.5 to 7 g/100 kcal of supplying energy to birds songs.
In the compositions of this invention can also include dietary fiber. Dietary fiber can be soluble or insoluble; as a rule, the preferred mixture of the fibers of these two groups. Acceptable IP is the full of dietary fiber include soy, peas, oats, pectin, guar gum, Arabia gum, fructo-oligosaccharides, galactooligosaccharides, sialyllactose and oligosaccharides derived from milk of animals. It is preferable to use a mixture of inulin with the short-chain fructo-oligosaccharides.
The compositions of the present invention can be obtained by any method known in the art. For example, if the composition is a nutrient mixture, as, for example, a mixture for feeding infants, it can be prepared by combining and mixing the correct ratio of protein source, a carbohydrate source and a source of fat. If required, the resulting mixture can be added emulsifiers. At this stage you can also add vitamins and minerals, but they usually include in the composition later in order to avoid their thermal destruction. Any lipophilic vitamins, emulsifiers and other compounds can be dissolved in the fat source before mixing it with other components. Then to achieve a liquid consistency can be added to water, preferably water purified by reverse osmosis.
The liquid mixture can be subjected to heat treatment to reduce bacterial load. For example, it can be quickly heated to a temperature of from about 120°to about 140°C and keep attemperator from about 5 to about 30 seconds. This procedure can be done through the injection of steam or by using a heat exchanger, for example a plate heat exchanger.
The liquid mixture is then cooled to a temperature of from about 60°to about 85°C; this can be done by the method of fast cooling. The obtained liquid mixture was then homogenized; it is, for example, in two stages (the first stage at a pressure of from about 7 MPa to about 40 MPa, in the second stage at a pressure of from about 2 MPa to about 14 MPa). Gomogenizirovannogo the mixture is then cooled to add heat-sensitive components, such as vitamins and minerals.
At this stage usually standardizes pH and dry matter content. Gomogenizirovannogo the mixture is placed in a suitable drying unit (for example, spray drying or freeze drying) and converted into a powder. The moisture content in the obtained powder should be no more than about 5 wt.%.
Probiotics for use according to the invention can be grown by any acceptable method and prepare for inclusion in the composition of the nutrient composition by, for example, by freeze drying or spray drying. Prepared probiotics can be added to the composition before it is subjected to heat treatment to reduce bacterial contamination. In the probiotics with innovatsia at least partially nerealizirane. Or probiotics separately subjected to a short heat treatment, so that they become nerealizirane, and then in liquid form or in powder form is included in the composition.
Selected probiotics can be grown by any suitable method and prepared for inclusion in the composition of the nutrient composition, for example, by freeze drying or spray drying. Or can be purchased from specialized manufacturers of bacterial preparations, already prepared in the desired form for inclusion in the composition of food products.
Exploring the immunogenic properties of living and past short-term high-temperature treatment of bacteria-probiotics, the authors present invention was determined by the ability of probiotics to induce the secretion of certain cytokines by human blood cells.
Immunogenic properties of viable probiotics compared with those of bacterial cells subjected to short-term high-temperature processing. It was found that the last heat treatment probiotics induce different levels of cytokine secretion than the corresponding live preparations.
Probiotics subjected to short-term exposure to high temperature, induced the secretion of smaller quantities of proinflammatory CIT is kinow (TNF-α, IFN-gamma, IL-12p40), at the same time maintaining or inducyruya more intense secretion of IL-10 than those living preparations. As a result, the ratio of IL-12p40/IL-10 in the case of the previous heat treatment of probiotics was lower than in the case of appropriate living cells. In contrast, bacteria are subjected to a temperature of 85°C for 20 minutes, causing higher levels of secretion of proinflammatory cytokines and lower levels of IL-10 than living cells; the result was to create a higher ratio of IL-12p40/IL-10. This indicates that in this invention a significant mode of heat treatment.
The ratio of IL-12p40/IL-10 produced by incubation of mononuclear cells in peripheral blood (RVMC) with probiotics in vitro, has predictive power in relation to anti-inflammatory actions in vivo (Foligne, B., et A1., 2007, World J. Gastroenterol. 13:236-243).
The present invention also relates to compositions according to the invention for the treatment or prevention of inflammatory disorders.
Accordingly, one of the embodiments of the present invention is the use of a composition according to the invention for the manufacture of a product intended for the treatment or prevention of inflammatory disorders.
What inflammatory disorders can be treated or prevented using compositions is s according to the invention, nothing specific is not limited.
For example, such disorders may be selected from the group consisting of acute inflammatory conditions (eg, sepsis); burns, chronic inflammatory processes, such as inflammatory bowel disease (Crohn's disease, ulcerative colitis, pucit); necrotizing enterocolitis; inflammatory skin diseases, such as inflammatory processes induced by UV radiation or chemical exposures, eczema, allergic inflammation of the skin; irritable bowel syndrome; inflammatory processes in the eyes; allergies and asthma; inflammatory processes associated with obesity; non-specific inflammatory processes associated with aging, and their combinations.
The present invention also encompasses a method of producing microorganisms, such as probiotic and/or starter cultures for dairy products, in particular viable probiotics and/or a viable starter cultures for dairy products that have anti-inflammatory action, or a way to strengthen anti-inflammatory action of microorganisms, in particular of viable probiotics and/or a viable starter cultures for dairy products, including the stage of short-term treatment of these microorganisms high t is mperature - at least 71,5°C for at least 1 second, for example, temperatures from about 71.5°C to about 150°C for a time from about 1 to about 120 seconds.
If viable probiotics and/or a viable starter cultures for dairy products are used in this way, short-term high-temperature processing causes at least part of the probiotic and/or starter cultures for dairy products is becoming rereplacenocase.
Short-term high-temperature processing can lead to transformation in rereplacenocase cells at least 90%, preferably at least 95%, more preferably at least 98%, most preferably at least 99%, preferably at least 99.9% of the, ideally, all of probiotic cells become nerealizirane.
In one of preferred embodiments of the present invention, this method includes a step of adding a viable probiotics in the composition according to the invention and short-term effects on probiotic composition of the high temperature.
The microorganisms according to this invention may be subjected to heat treatment before inclusion in the composition of the product.
The composition of this invention can be any; for example, it may be food or PI is athelny product or beverage, which includes probiotics.
This invention also features a method of obtaining a composition containing microorganisms, such as probiotic and/or starter cultures for dairy products, preferably viable probiotics and/or a viable starter cultures for dairy products that have anti-inflammatory properties, or improve existing anti-inflammatory properties, including the stage of short-term high-temperature processing of these microorganisms with at least 71,5°C for 1 second, for example at a temperature of from about 71.5°C to 150°C for a time from about 1 to about 120 seconds.
Stage short high temperature treatment can be carried out using industrial equipment, but also at home by using, for example, the steam generator. In the latter case, anti-inflammatory product can be purchased immediately prior to its consumption.
Specialists in the art it should be clear that it is possible to freely combine all the described features of the present invention without going beyond its scope disclosed herein. In particular, the features related to the use and method according to the invention, applicable to methods and compositions according to of the briteney and Vice versa.
Other advantages and features of the present invention follow from the following examples and illustrations.
Brief description of figures
Figures 1A and 1B illustrate the increased anti-inflammatory properties of probiotics, subjected to short-term exposure to high temperature.
Figure 2 illustrates the fact that not possessing anti-inflammatory properties of probiotic strains after a short high temperature treatment buy anti-inflammatory properties, and that they have a pronounced anti-inflammatory immunogenic profile in vitro,
Figures 3A and 3B illustrate the fact that the probiotic strains used in commercially available products, after short-term high-temperature processing are enhanced or new immunogenic anti-inflammatory properties in vitro,
Figures 4A and 4B illustrate the fact that the strains of starter cultures for dairy products (for example, Lc1) after heat treatment at high temperature are enhanced or new immunogenic anti-inflammatory properties in vitro.
Figure 5 illustrates the fact that not possessing anti-inflammatory properties of probiotic strains after treatment type HTST show immunogenic anti-inflammatory properties in vitro.
Figure 6 depicts achiev Italy of principal component analysis of the data, obtained in experiments with mononuclear cells of peripheral blood (RVMS), which secretively cytokines (IL-12p40, IFN-γ, TNF-α, IL-10) under the influence of strains of probiotics and starter cultures for dairy products, previous heat treatment (at a temperature of 140°C for 15 seconds) or in native form. Each symbol (circle or square) corresponds to one strain (native or past heat treatment) and is identified by a registration number in the culture collections of the Nestle company (NCC) or the name of this strain (italic).
Figure 7 shows the relationship of IL-12p40/IL-10 for native and previous heat treatment (at a temperature of 85°C for 20 minutes) strains. In General, heat treatment at a temperature of 85°C for 20 minutes leads to the increase of the ratio of IL-12p40/IL-10 in contrast to short-term heat treatment according to this invention (figures 1, 2, 3, 4 and 5).
Obtaining bacterial medications
It is generally considered that a healthy effect on the immune system of the host body, provided live probiotics, is stampartic. It was shown that probiotic strains that induce high levels of Il-10 and/or low levels of proinflammatory cytokines in vitro (definition in the system with RVMS) are powerful is passed anti-inflammatory agents in vivo (Foligne, C., et aL, 2007, World J. Gastroenterol. 13:236-243).
To study the anti-inflammatory properties of probiotics after heat treatment was used the following strains: Bifidobacterium longum NCC 3001, Bifidobacterium longum NCC 2705, Bifidobacterium breve NCC 2950, Bifidobacterium lactis NCC 2818, Lactobacillus paracasei NCC 2461, Lactobacillus rhamnosus NCC 4007, Lactobacillus casei NCC 4006, Lactobacillus acidophilus NCC 3009, Lactobacillus casei ACA-DC 6002 (NCC 1825) and Escherichia coli Nissle. Was also tested some strains of starter cultures for dairy products used for the manufacture of products of lactic acid fermentation in Nestle containing probiotic Lc1: Streptococcus thermophilus NCC 2019, Streptococcus thermophilus NCC 2059, Lactobacillus bulgaricus NCC 15 and Lactococcus lactis NCC 2287.
Bacterial cells were cultured in bioreactors of 5-15 l under conditions optimized for each of the strains. Used all normally used for culturing bacteria culture medium. Such environments are known to specialists in this field of technology. When the pH reached 5.5 continuously added a 30% solution of alkali (NaOH or Ca(OH)2). When required anaerobic conditions, the free space of the reactor was filled with CO2. Cells of E. coli were cultivated under standard aerobic conditions.
Bacterial cells were collected by centrifugation (5000 g at 4°C) and resuspendable in phosphate buffer solution (PBS) in a volume to obtain the final concentration is approximately 10 9-1010CFU/ml. part of the preparations were frozen at 80°C in 15% glycerol. The remaining cells were subjected to heat treatment as follows:
- Very high temperature (UHT): 140°C for 15 seconds; indirect injection of steam;
- High temperature short time (HTST): 74°C, 90°C and 120°C for 15 seconds; indirect injection of steam;
- Relatively low temperature for a relatively long time: 85°C for 20 minutes in a water bath.
After heat treatment the samples were frozen and kept at -80°C until use.
The definition of immunogenic properties of the bacterial preparations in vitro.
Defined immunogenic properties of living and subjected to heat treatment bacterial preparations, namely their ability to induce the secretion of certain cytokines by human blood cells in vitro. Mononuclear cells from peripheral blood (RVMS) was isolated from the cell material after filtration of the blood. After separation of cells in a gradient density mononuclear cells were collected and washed twice with balanced salt solution and Hank. The cells are then resuspendable in the environment Dulbecco modified according to the method Iscove (IMDM, Sigma), which was added fetal calf serum 10% (Bioconcept, Paris, France), 1% L-glutamine (Sigma), 1% penicillin/streptomycin (Sigma) and 0% gentamicin (Sigma). Then the samples RVMS (7×105cells per well) were incubated with live and subjected to heat treatment by bacteria (in an amount equivalent to 7×10 CFU per well) in 48-hole tablets within 36 hours. Determined the impact of living and subjected to heat treatment of bacteria on RVMS from 8 different donors in two separate experiments. After 36 hours of incubation, the culture, the tablets were frozen and kept at -20°C until determination of cytokines. The profile of cytokines was determined in parallel (i.e. in the same experiment with the same party RVMS) for live bacteria and subjected to heat treatment such as bacteria,
The levels of cytokines - interferon-gamma (INF-γ), interleukin-l (IL-12p40), alpha tumor necrosis factor (TNF-α) and interleukin-10(IL-10) was determined in the supernatant of culture medium after 36 hours of incubation by enzyme immunoassay ELISA (human IL-10 R&D DuoSet human IL-12p40 BD OptEIA human TNF-α BD OptEIA human INF-γ BD OptEIA), following the manufacturer's instructions. Gamma-interferon, interleukin-R and alpha tumor necrosis factor are proinflammatory cytokines, and interleukin-10 is a powerful anti-inflammatory agent. The results of the experiments were expressed as mean value (PG/ml)+/- standard error of the mean (SEM) for 4 different donors; performance is established with representation from two separate experiments with four donors each. For each strain was calculated the ratio of IL-12p40/IL-10, have predictive power with respect to the anti-inflammatory effect in vivo (Foligne, B., et A1., 2007, World J. Gastroenterol. 13:236-243).
Obtained for each strain numerical values of the levels of cytokines (PG/ml), as defined by the ELISA method (see above) substituted in the program BioNumerics v5.10 (Applied Maths, Smt-Martens-Latem, Belgium). This array data was performed by the principal component analysis (PCA), including subtraction of mean and division by the variance.
Anti-inflammatory properties that are created ultra-high-temperature (UHTV short-term high-temperature (HTST) processing
Studied probiotic strains were subjected to the processing of very high temperature (of ultra-pasteurization - UHT), short-term high temperature processing (HTST) and temperature 85°C for 20 minutes and compared their immunogenic properties with those of living cells in vitro. During incubation with human mononuclear cells of peripheral blood (RVMS) living microorganisms (probiotic and/or starter cultures for dairy products) induced different levels of cytokine secretion (Figures 1, 2, 3, 4 and 5). In the heat treatment of these organisms varied levels of cytokines produced RUMS, and dependent on temperature. After krakowieniezarejestrowana treatment (120°C or 140°C for 15 seconds) was obtained rereplacenocase bacteria, it has anti-inflammatory immunogenic properties (figures 1, 2, 3 and 4). The strains subjected to UHT treatment (140°C, 15 s), induced less intense secretion of proinflammatory cytokines (TNF-α, IFN-γ, IL-12p40), at the same time maintaining or inducyruya additional (compared to the live agents in the formation of IL-10. As a result, for all strains treated UHT, values, attitudes IL-12p40/IL-10 were lower than for living cells (figures 1, 2, 3 and 4). This observation is also true for bacteria treated HTST, namely a temperature of 120°C for 15 s (figures 1, 2, 3 and 4) or 74°C and 90°C for 15 s (figure 5). Various heat treatment (UHT or HTST) had a similar effect on the immunogenic properties in vitro probiotic strains (figures 1, 2, 3 and 5) and starter cultures for dairy products (figure 4). The principal component analysis results of changes in the secretion of cytokines by cells RUMS under the influence of the living and the previous heat treatment (140°C, 15 s) probiotic strains and starter cultures for dairy products showed that all living strains are distributed on a scale of "x" (figure 6); this shows that in vitro bacteria induce the secretion of proinflammatory cytokines in very different ways - from low (left) to high (right side). In this graph, strains, proseds the e heat treatment, group left that demonstrates a weak induction of proinflammatory cytokines. In contrast, bacteria are subjected to a temperature of 85°C for 20 minutes, induced the secretion of large quantities of Pro-inflammatory cytokines and less IL-10 than living cells; the result was a higher value relationships of IL-12p40/IL-10 (Figure 7).
Anti-inflammatory properties are enhanced or created as a result of processing type UHT and HTST.
Strains, processed UHT and HTST, show anti-inflammatory properties independently from the respective source (living cells) immunogenic properties. It is shown that the probiotic strains that have anti-inflammatory properties in vivo and exhibiting anti-inflammatory properties in vitro (B. longum NCC 3001, B. longum NCC 2705, B. breve NCC 2950, B. lactis NCC 2818), after a short high temperature treatment anti-inflammatory properties in vitro amplified. As figure 1 illustrates, for strains of Bifidobacterium treated UHT, the ratio of IL-12p40/IL-10 was lower than for the corresponding living cells; it exhibits improved anti-inflammatory properties of the samples subjected to UHT processing. Even more interestingly, also confirmed the emergence of the anti-inflammatory properties in the treatment UHT and HTST have not possessed anti-inflammatory with what properties of living strains. For live cells of L. rhamnosus NCC 4007 and L paracasei NCC 2461, had a high ratio of IL-12p40/IL-10 in vitro (figures 2 and 5). It was shown that in mice, these two strains do not protect from colitis induced by TNBS, L rhamnosus NCC 4007 and L. paracasei NCC 2461 ratio of IL-12p40/IL-10 was significantly decreased after short-term high-temperature processing (UHT or HTST), reaching the same low level as in the case of Bifidobacterium strains. These low IL-12p40/IL-10 due to low secretion of IL-R in combination with unchanged (L. rhamnosus NCC 4007) or significantly higher degree of induction of the secretion of IL-10 (L. paracasei NCC 2461) (figure 2).
- Anti-inflammatory properties of living organisms can be strengthened by heat treatment type UHT and HTST (e.g., B. longum NCC 2705, B. longum NCC 3001, B. breve NCC 2950, B. lactis NCC 2818);
- Anti-inflammatory properties can occur in living organisms, not possessing anti-inflammatory effect (e.g., L. rhamnosus NCC 4007, L. paracasei NCC 2461, starter culture of S. thermophilus NCC 2019) as a result of heat treatment type UHT and HTST;
- Anti-inflammatory properties are also demonstrated in strains isolated from commercially available products (figures 3A and B), including probiotic strain of E. coli.
The influence of treatment type UHT/HTST was similar for all the tested probiotics and starter cultures for dairy products, n is an example of lactobacilli, bifidobacteria and streptococci.
Processing type UHT/HTST applied to a number of lactobacilli, bifidobacteria and streptococci, showing different immunogenic properties in vitro. After processing type UHT/HTST all strains induced a lower secretion of proinflammatory cytokines than those living cells (figures 1, 2, 3, 4, 5 and 6); this demonstrates that the influence of treatment type UHT/HTST on the immunogenic properties of the resulting rereplacenocase bacteria can be attributed to all the probiotics, in particular lactobacilli and bifidobacteria, as well as certain strains of E. coli and all starter cultures for dairy products, in particular Streptococcus, the lactococcal and lactobacilli.
1. Composition for prevention or treatment of inflammatory disorders, containing microorganisms subjected to heat treatment at 120-140°C for a short period of time of 5-15 seconds, in which the microorganisms are selected from the group consisting of probiotics, starter cultures lactic cultures or combinations thereof, where the probiotic and/or starter culture dairy cultures are selected from the group consisting of Bifidobacterium longum NCC 3001, Bifidobacterium longum NCC 2705, Bifidobacterium breve NCC 2950, Bifidobacterium lactis NCC 2818, Lactobacillus paracasei NCC 2461, Lactobacillus rhamnosus NCC 4007, Streptococcus thermophilus NCC 2019, Streptococcus thermophilus NCC 2059, Lactobacillus casei NCC 4006, Lactobacillus casei ACA-DC 6002 (NCC 1825), Lactobacillus acidophilus NCC 3009, Escherichia coli Nissle, Lactobacilus bulgaricus NCC 15, Lactococcus lactis NCC 2287 or combinations thereof.
2. Composition under item 1, in which at least 90%, preferably at least 95%, more preferably at least 98%, most preferably at least 99%, preferably at least 99.9% of the, ideally, all of probiotic cells and/or cells of sourdough lactic cultures are nerealizirane.
3. The composition according to p. 1, which can be introduced into the body orally, enterline, parenterally, for example subcutaneously or intramuscularly, intrawaginalno intrarectal, locally or intraocular.
4. Composition according to any one of paragraphs. 1-3, designed for people or Pets.
5. The composition according to p. 2, which contains from about 0.005 mg to about 1000 mg rereplacenocase of organisms per daily dose.
6. Composition under item 1, in which the inflammatory disorder is selected from the group consisting of acute inflammatory conditions, such as sepsis, burns, chronic inflammatory processes, such as inflammatory bowel disease (such as Crohn's disease, ulcerative colitis, pucit); necrotizing colitis; irritable bowel syndrome; inflammatory skin lesions, such as inflammation caused by exposure to ultraviolet radiation or chemicals, eczema, allergic skin inflammation; inflammation is processov in the eyes; allergies, asthma; inflammatory processes associated with obesity; non-specific inflammatory processes associated with aging, and their combinations.
7. The method of receiving probiotics and/or dairy starter cultures cultures with anti-inflammatory effect or to enhance the anti-inflammatory effect of probiotics and/or sourdough lactic cultures, including the state of exposure of probiotic and/or starter cultures lactic cultures to high-temperature treatment at 120-140°C for a short period of time of 5-15 seconds, in which the probiotic and/or starter culture dairy cultures are selected from the group consisting of Bifidobacterium longum NCC 3001, Bifidobacterium longum NCC 2705, Bifidobacterium breve NCC 2950, Bifidobacterium lactis NCC 2818, Lactobacillus paracasei NCC 2461, Lactobacillus rhamnosus NCC 4007, Streptococcus thermophilus NCC 2019, Streptococcus thermophilus NCC 2059, Lactobacillus casei NCC 4006, Lactobacillus casei ACA-DC 6002 (NCC 1825), Lactobacillus acidophilus NCC 3009, Escherichia coli Nissle, Lactobacillus bulgaricus NCC 15, Lactococcus lactis NCC 2287 or combinations thereof.
8. The method according to p. 7, in which short-term high-temperature treatment leads to the conversion of at least 90%, preferably at least 95%, more preferably at least 98%, most preferably at least 99%, preferably at least 99.9% of the, ideally, all of probiotic cells and/or cells of sourdough lactic cultures in rereplacenocase.
9. A method of obtaining a composition containing probiotics and/or is aquasky dairy cultures, with anti-inflammatory properties, or to improve existing anti-inflammatory properties, including the state of exposure of probiotic and/or starter cultures lactic cultures to high-temperature treatment at 120-140°C for a short period of time of 5-15 seconds, in which the probiotic and/or starter culture dairy cultures are selected from the group consisting of Bifidobacterium longum NCC 3001, Bifidobacterium longum NCC 2705, Bifidobacterium breve NCC 2950, Bifidobacterium lactis NCC 2818, Lactobacillus paracasei NCC 2461, Lactobacillus rhamnosus NCC 4007, Streptococcus thermophilus NCC 2019, Streptococcus thermophilus NCC 2059, Lactobacillus casei NCC 4006, Lactobacillus casei ACA-DC 6002 (NCC 1825), Lactobacillus acidophilus NCC 3009, Escherichia coli Nissle, Lactobacillus bulgaricus NCC 15, Lactococcus lactis NCC 2287 or combinations thereof.
10. The method according to p. 9, which provides for the addition of viable probiotic and/or starter cultures lactic cultures in the composition and exposure of the composition containing the probiotic and/or starter culture dairy cultures, short-term high-temperature processing at 120-140°C for 5-15 seconds.
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to new compounds of formula I, such as below, or its pharmaceutically acceptable salts. What is described is a method for preparing them.
wherein: A independently from B means phenyl,
, or ,
and B independently from A means phenyl,
, or ,
and the values Z, Y, D, L1, L2, L3, Z1, Z2 are presented in the patent claim.
EFFECT: compounds are effective for hepatitis C virus (HCV) replication inhibition.
17 cl, 3 tbl, 8 dwg, 177 ex
SUBSTANCE: group concerns using undecapeptide - H-Tyr-Pro-D-Arg-D-Arg-D-Arg-D-Arg-D-Arg-D-Arg-D-Arg-D-Arg-Gly-OH·9HCl as an analgesic agent. The group of inventions also concerns a dosage form containing the above peptide as an active substance for treating acute and chronic pain syndromes.
EFFECT: stable and prolonged analgesic effect both in the enteral, and oral route of administration.
3 cl, 2 ex, 2 dwg, 5 tbl
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention can be used in chemical-pharmaceutical industry for preparing effective tranquilising salamide preparations and analgesics. What is presented is using salicyloyl morpholine sodium salt of formula (I) as a tranquilising, nootropic and analgesic agent possessing low gastric toxicity.
EFFECT: implementing the declared application with the therapeutic index of salicyloyl morpholine of formula (I) is 3 times higher than that of mexidole, and 9,5 times higher than that of aspirin.
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention relates to field of organic chemistry, namely to heterocyclic compound of general formula (I) or to its pharmaceutically acceptable salt, acid salt or stereoisomer, where Y: NRa and N+R1R2X-; Z: bond, -(CH2)p, -CHOH, -CH=CH-, -C≡C-, -CONH- and -CO-; Rb: C1-C8 alkyl, C2-C8 alkenyl, C6-C10 aryl, -NR5R6,: , and ; with each alkyl, alkenyl and aryl, representing Rb, possibly, contains 1-3 substituents, selected from C1-C4 alkyl, C2-C4 alkenyl, C3-C6 cycloalkyl, C1-C4 alkoxy, C6-C10 aryl, 5-, 6- and 7-membered heterocyclyl, containing 1-3 heteroatoms, selected from nitrogen, oxygen and sulphur, halogen, -OH, -NH2, -CN and -NO2; Rc: halogen, C1-C6 alkyl, C2-C6 alkenyl, C2-C6 alkinyl, C3-C10 cycloalkyl, C3-C8 cycloalkenyl, C1-C4 alkoxy, C6-C10 aryl, 5-, 6-, 7- and 8-membered monocyclic heterocyclyl, containing 1-3 heteroatoms, selected from nitrogen, oxygen and sulphur, 9- and 10-membered bicyclic heterocyclyl, containing 1-3 heteroatoms, selected from nitrogen, oxygen and sulphur; with C1-C6 alkyl, C2-C6 alkenyl C2-C6 alkinyl, C3-C10 cycloalkyl, C3-C8 cycloalkenyl, C6-C10aryl, 5-, 6-, 7-, 8-membered monocyclic heterocyclyl and 9- and 10-membered bicyclic heterocyclyl, representing Rc, possibly contain 1-5 substituents, selected from the group, consisting of C1-C4 alkyl, C1-C4 alkoxy, C1-C4 haloalkyl, C1-C4 haloalkoxy, C3-C6 cycloalkyl, C4-C8 cycloalkenyla, halogen, -OH, -NH2, C6-C10 (A)(A')(A")(A'")aryl, (A)(A')(A")(A'")heterocyclyl, containing 1-3 heteroatom, selected from nitrogen, oxygen and sulphur, NR14R15, (CH2)pNR14R15, -CN, -NO2, oxo, -COOR14, SOR14, SO2R14, SO2NR14R15, NR15SO2R16, COR14, CONR14R15 and NR15COR16; with each (A), (A'), (A") and (A'")independently absent or representing C1-C4 alkyl, and each heterocyclyl (A)(A')(A")(A'")heterocyclyl is independently selected from the group, consisting of 5-, 6-, 7- and 8-membered monocyclic heterocyclyl, containing 1-3 heteroatoms, selected from nitrogen, oxygen and sulphur, and 9- and 10-membered bicyclico heterocyclyl, containing 1-3 heteroatoms, selected from nitrogen, oxygen and sulphur; the remaining radicals have values given in i.1;and on condition that, if Rc represents heterocyclyl, said heterocyclyl is bound directly through carbon atom of heterocyclyl ring. Invention also relates to particular compounds and to pharmaceutical composition based on formula (I) compound.
EFFECT: obtained are novel imidazopyrazine and imidazodiazepine derivatives, useful for prevention or treatment of disease or condition, severity of which is reduced by receptors to cannabinoids.
21 cl, 5 dwg, 4 tbl, 71 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to compounds of formula (I) - oxytocin receptor agonists, to their pharmaceutical compositions containing them.
EFFECT: compounds can be used for preparing a drug for treating, among others, abdominal pain, irritable bowel syndrome (IBS), autism, erectile dysfunction, female sexual dysfunction, delivery stimulation and maintenance, lactation stimulation and maintenance, post-delivery syndrome, posttraumatic stress disorder (PTSD), pain, anxiety and other conditions.
19 cl, 1 tbl, 5 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: present invention refers to isoxazoline FAAH inhibitors of formula (I) or their pharmaceutically acceptable forms, wherein each of G, Ra, Rb, Rc and Rd has a value described in the present application, to pharmaceutical compositions, and methods of treating a FAAH-mediated condition.
EFFECT: developing the method of treating the FAAH-mediated condition.
32 cl, 22 tbl, 351 ex
SUBSTANCE: in claimed co-crystalline form molar ratio of 2-hydroxybenzamide and salicylic acid constitutes 1:1. Co-crystalline form is characterised by peaks at 2θ(°) 11.5, 14.8, 16.7, 18.8, 22.2, 23.5, 27.0 by the data of measurement of X-ray radiation diffraction on powder, and has endothermic peak from 116 to 124°C by the data of measurement by means of differential scanning calorimetry. Claimed co-crystalline form can be applied for manufacturing pharmaceuticals.
EFFECT: increased solubility of 2-hydroxybenzamide.
7 dwg, 2 ex
FIELD: food industry.
SUBSTANCE: biologically active food additive strengthening the organism adaptive power and body defences and having anti-inflammatory and antioxidant activity contains vegetal origin components represented by a complex extract of devil's-club root, Rhaponticum carthamoides root, Hedysarum neglectum Ledeb root, celery roots and leaves, rhodiola rosea root, Japanese angelica tree roots, boschniakia rossica roots, Hungarian sainfoin herb, magnolia-vine fruits; additionally the additive contains chitosan, trepang fermentative hydrolysate, ascorbic acid, taurine, glutathione, nicotinamide, vitamin B1, vitamin B2, vitamin B6, vitamin B12, folic acid, anhydrous calcium chloride, magnesium chloride, zinc chloride, bee honey at preset ingredients ratio.
EFFECT: biologically active food additive promotes effective strengthening of the organism adaptive power and body defences and human aging retardation.
4 tbl, 6 ex
SUBSTANCE: invention relates to medication, representing betulin bis-isonicotinoate .
EFFECT: low toxicity, expressed anti-ulcer, anti-inflammatory and hepatoprotective activity.
3 tbl, 5 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to compounds of formula I wherein R means C1-6alkyl, C1-6halogenalkyl, hydroxy-C1-6alkyl, hydroxygroup or halogen; m, n is equal to 0 or 1; Z1 means CH or NH; Z2 means CH or N; Z3 means CR1, N or NR2; R1 means H, C1-6alkyl, C3-7cycloalkyl, cyanogroup, cyano-C1-6alkyl or halogen; R2 means H or C1-6alkyl; X means CH, CR' or N; X' means CH, CR' or N; r is equal to 1; Y means CH or CR'; R' means R'a or R'b; R'a means a halogen or cyanogroup; R'b means C1-6alkyl, heterocycloalkyl specified in piperazinyl, morpholinyl, piperidinyl, thiomorpholinyl, azetidinyl, pyrrolidinyl, OR", SR", S(=O)2R" or NR"R", optionally substituted by one or more R'c; R'c means a hydroxygroup, oxogroup, cyanogroup, C1-6alkyl, pyridinyl, carboxy-C1-6alkyl, aminocarbonyl-C1-6alkylaminogroup, C1-6alkylaminogroup, C1-6dialkylaminogroup or C1-6alkoxygroup; R" means H, C1-6alkyl, hydroxy-C1-6alkyl, piperidinyl, C3-7cycloalkyl or pyridinyl; Q means S(=O)2Q1, C(=O)Q2, C(=O)OQ3 or Q4; Q1 means C1-6alkyl, C3-7cycloalkyl-C1-6alkyl, C1-6alkylaminogroup or C1-6dialkylaminogroup optionally substituted by one or more Q1'; each Q1' independently means C1-6alkyl or cyanogroup; Q2 means C1-6alkyl optionally substituted by one or more Q2'; each Q2' independently means a cyanogroup; Q3 means C1-6alkyl; Q4 means C1-6alkyl, oxetanyl optionally substituted by one or more Q4'; each Q4' independently means a halogen, cyanogroup, cyano-C1-6alkyl; p is equal to 0, 1 or 2; q is equal to 1 or 2; each means a single bond or a double bond; provided one of Z1 and Z2, and Z3 and Z3 bonds are double and single.
EFFECT: compounds of formula I as JAK inhibitors.
23 cl, 2 tbl, 121 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention relates to industrial microbiology, namely to application of strain of Lactobacillus casei, deposited under number I-1518, for obtaining composition for stimulating response of cells-natural killers in adult individual with common respiratory infectious disease, where composition also contains bacteria Lactobacillus bulgaricus and Streptococcus thermophilus.
EFFECT: application of composition, containing Lactobacillus casei I-1518, makes it possible to increase resistance of organism to common respiratory infections, in particular to rhinopharyngitis or laryngitis.
17 cl, 12 tbl, 1 dwg, 3 ex
FIELD: food industry.
SUBSTANCE: inventions group relates to biotechnology and may be used for preparing bacterial preparations applicable as probiotic biologically active additives. The bacterial concentrate production method envisages preparation of a nutritional medium, sterilisation and cooling. Inoculum introduction, cell propagation, bacterial mass separation from culture liquid, dispensing and closuring. Into the nutritional medium composition one introduces cedar or linseed oil of fish or ringed seal fat in an amount of 1-1.5% of the medium weight; the inoculum is represented by a Bifidobacterium longum DK-100 strain. The produced bacterial concentrate is used as a probiotic biologically active food additive.
EFFECT: inventions group allows to recover the digestive tract microflora.
2 cl, 3 tbl, 3 dwg, 5 ex
SUBSTANCE: invention relates to biotechnology and medical industry and can be used in production of bacterial concentrates, biologically active food additives, fermented food products. Method of obtaining bacterial concentrate of bifidobacteria in liquid form includes preparation of nutritional medium with addition of growth components based on clarified cottage cheese whey or on water base with addition of up to 1.5% of glucose, or on soybean whey with addition of lactose in amount 1%. Strain of bifidobacteria B. bifidum 83, activated with β-halactosidase, is introduced into prepared nutritional medium in amount 3-5%, biomass is grown, cooled, poured in containers.
EFFECT: invention makes it possible to increase manufacturability and intensity of the process, increase biochemical activity of bifidobacteria and consumer properties of obtained product.
7 tbl, 5 dwg, 4 ex
FIELD: food industry.
SUBSTANCE: invention relates to infant alimentation field. The infant nutritional mixture, administered to a baby as a single source of nutrition or a single additional source of nutrition as an addition to breast feeding, contains non-replicated probiotic microorganisms brought to a non-replicated condition by way of 71.5-150°C high-temperature treatment during 1-120 sec.
EFFECT: invention allows to produce a nutritional mixture with improved anti-inflammatory immunogenic properties.
15 cl, 12 dwg, 6 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: group of inventions relates to biotechnology. Claimed is strain of Lactobacillus delbrueckii subspecies lactis CNCM I-3741, reducing content of cholesterol in blood. Strain is applied for obtaining fermented dairy products.
EFFECT: inventions provide efficient reduction of cholesterol content in blood.
3 cl, 3 dwg, 2 ex
SUBSTANCE: group of inventions relates to biotechnology. Disclosed is a Bacillus subtilis VKPM V-11353 strain, capable of splitting a wide range of mono- and di-sugars and a wide range antagonistic effect on pathogenic and opportunistic pathogenic bacteria and fungi, which cause diseases in plants and farm animals. Also disclosed are versions of using the Bacillus subtilis VKPM V-11353 strain as a bacterial preservative for silos, for producing agents for normalising intestinal microflora of animal farms and for producing agents for protecting plants from diseases.
EFFECT: group of inventions improves the quality of silos, improves safety of animal farms and increases effectiveness against pathogenic and opportunistic pathogenic microorganisms which cause wheat diseases.
4 cl, 8 tbl, 4 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to microbiology. What is presented is using the strain Bifidobacterium longum NCC 2705 (CNCM-I2618) for preparing a complete nutrient composition used for relieving symptoms of allergy to food in the patients suffering allergies caused by ingestant allergens.
EFFECT: composition provides the secondary prevention of allergic reactions caused by the above ingestant allergens.
16 cl, 7 dwg, 1 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to medicine and concerns a formulation for treating skin damages containing a therapeutic agent and an ointment base. The therapeutic agent is presented by a microbial cell suspension of nonpathogenic plasmid-free strain E. coli, producing human recombinant peroxiredoxin 6 identical in an amino acid sequence to natural protein. The ointment base contains Vaseline, lanoline, normal saline and phenol as a preserving agent.
EFFECT: invention provides the active skin penetration of the ingredient of the therapeutic agent that enhances the therapeutic action and reduces the length of treatment.
3 ex, 1 tbl
FIELD: veterinary medicine.
SUBSTANCE: method comprises intravaginal administration of probiotic preparation "Giprolam" (Lactobacillus fermentum 44/1 and Lactococcus lactis subsp. Lactis 574). "Giprolam" is administered for 5-7 days prior to calving daily at a dose of 100 cm3.
EFFECT: use of the claimed invention promotes to maintain the amount of lactobacilli and bifidobacteria at the physiological level, prevents the colonisation of the genital tract with potentially pathogenic microflora and prevents the occurrence of postpartum infections.
6 tbl, 2 ex
SUBSTANCE: analgesic preparation causing medication overuse headache are withdrawn; a detoxification therapy is conducted; an analgesic alternative is provided; a preventive medication and a behaviour therapy are applied. The preventive medication represents single intramuscular administration of xeomin into temporal muscles in a dose of 25 units in each frontal and occipital muscle in a dose of 12.5 units. The preventive medication follows the detoxification therapy.
EFFECT: considerable reduction of the recurrence number by relieving the reflex pericranial muscle and expression line tension.
FIELD: biotechnology, microbiology, medicine.
SUBSTANCE: invention relates to the strain Lactobacillus paracasei CNCM I-2116 used for diarrhea prophylaxis causing by pathogenic microorganisms. Supernatant of this strain culture elicits ability to prevent colonization of intestine with pathogenic microorganisms causing diarrhea also and this strain is designated for preparing agent used for prophylaxis and/or treatment of disorders associated with diarrhea. Agent for oral administration represents therapeutically effective dose of the strain L. paracasei CNCM I-2116 or supernatant of its culture and acceptable foodstuff. Invention provides the enhanced viability of the strain in its applying and effectiveness in prophylaxis of adhesion to intestine cells and invasion to intestine cells of pathogenic microorganisms causing diarrhea.
EFFECT: valuable medicinal properties of strain.
5 cl, 8 dwg, 10 ex