Method of determining compositions stimulating skin stem cell formation
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to medicine. A composition for stimulating the skin stem cell production containing interleukin-1 alpha and a dermatologically acceptable diluent or carrier.
EFFECT: invention provides improving the stem cell stimulation.
The technical field
 This invention relates to a drug designed to stimulate production of stem cells in the skin, and the method of their identification.
The level of technology
 a Stem cell is a cell that has two properties: (1) retains the ability to heal itself by dividing mitotic cells while maintaining the undifferentiated state and (2) has activity to differentiate into any type of specialized cell. Stem cells can be identified by specific markers of stem cells. Such markers are well known in the prior art (Rao R, et al., Biology of Reproduction, 71: 1772-1778, 2004). Usually markers of stem cells represent gene products that functionally linked to the maintenance of the undifferentiated state of stem cells, maintaining prepotential, multipotential or sometimes unipotential. An example of a reliable marker of stem cells is HOST, which is often called OSC/4 or Pou5f1. It is a transcription factor of the POU family is very important to ensure the undifferentiated state of stem cells and prepotential (Watson CM, et al., Cell Struct. Func 90, 26: 123-129, 2001; Pesce M et al.. Stem Cells, 19: 271-278, 2001).
 In Mature skin revealed at least three populations of endogenous stem cells. Population of stem cells the detection is provided in the convex part of the hair follicle (Potten CS et al. Experientia 39, 1125-1129, 1983; Watt FM. Philos. Trans. R. Soc. Lond. B.Biol. Sci. 353, 831-837, 1998; Lavker RM et al. Proc. Natl. Acad. Sci. USA. 97, 13473-13475, 2000). It is known that another population of stem cells located in mezhvolokonnom basal layer of the epidermis (Tumbar T et al. Science 303, 359-363, 2004; Taylor G et al. Cell 102, 451-461, 2000). Another population of stem cells is a mesenchymal cell. This population is able to form many cell types in the laboratory and even capable of generating layer of the epidermis composed of cells of different types in the 3-dimensional skin model (Crigler L, et al.. FASEB Journal. 2007 21: 2050-2063). Because of the ability of endogenous stem cells for skin regeneration there is a need for substances that can stimulate the production of stem cells in the skin to activate skin regeneration and renewal process under adverse circumstances, such as injury, illness or aging. However, reliable identification of such substances is limited by the lack of suitable methods.
 the Purpose of this invention is to provide a medicine designed to stimulate production of stem cells in the skin, and the method of their identification.
Detailed description of the invention
 This invention provides a method for identifying a drug candidate designed to stimulate production of stem cells in the skin include the s: (a) the application of the test drug on the skin of the animal; and (b) determining, improving test drug level markers of stem cells in the skin of the animal, identifying thus the test drug as intended to stimulate production of stem cells in the skin, if the test drug increases levels of markers of stem cells in the skin.
 additionally, this invention provides a drug designed to stimulate production of stem cells in the skin, with the specified drug is identified by the method comprising: (a) the application of the test drug on the skin of the animal; and (b) determining, improving test drug level markers of stem cells in the skin of the animal, identifying thus the test drug as a drug designed to stimulate production of stem cells in the skin, if the test drug increases levels of markers of stem cells in the skin.
 additionally, this invention provides a composition for stimulating the production of stem cells in the skin, containing a drug designed to stimulate production of stem cells in the skin, with the specified drug is identified by the method comprising: (a) the application of the test drug on the skin of the animal; and (b) determining, improving test drug level markers of stem cells in the skin zivotnog is, identifying thus the test drug as a drug designed to stimulate production of stem cells in the skin, if the test drug increases levels of markers of stem cells in the skin; and acceptable for cosmetic purposes diluent or carrier.
 Preferably, the animal according to the present invention has been mammals. More preferably, the mammal was a mouse.
 the Term "stem cell" refers to a cell that has two properties: (1) regeneration and (2) activity, where the term self-healing refers to the ability of cells to undergo numerous cycles of cell division while maintaining the undifferentiated state and where the term activity refers to the ability of cells to differentiate into any type of specialized cells.
 Preferably, the stem cells according to this invention were poly potent, multipotent or unipotent stem cells. More preferably, the stem cells according to this invention were poly potent stem cells. The term poly potent refers to a stem cell that can differentiate into cells derived from any of the three layers of the skin, namely, the mesoderm, endoderm and the ectoderm, from which arise all cells of the body. The term "multi-pot the percentage" refers to a stem cell, that can occur only cells of a closely-related family of cells. The term "unipotent" refers to a stem cell that can differentiate only into cells of the same type.
 the Term "stem cell marker" refers to any marker that recognizes poly potent, multipotent or unipotent cells in the skin. This invention is not limited anyhow specific marker for stem cells, but applies to all such markers, currently known or open or hereafter developed. Specialist in the art knows how to identify specific suitable marker of stem cells for use in the method according to this invention. However, the preferred marker of stem cells according to this invention is selected from the group of marker gene expression, consisting of
More preferably, the stem cell marker according to this invention was OCT. In the practical application of this invention, the stem cell marker can be detected by the method known from the prior art. Such methods include, inter alia, Northern blotting, Western blotting, immunohistochemistry, flow cytometry, ELISA analysis with the enzyme label and immune electron spectroscopy.
 the Term "germ teologichesky acceptable diluent or carrier" refers to one or more compatible solid or liquid diluents or carriers of cells fillers, suitable for introduction into any part of the human skin and is compatible with the product suitable to stimulate production of stem cells. Examples of such carriers include, inter alia, distilled or deionized water, propylene glycol, glycerin and oil.
 Preferably, the drug is suitable to stimulate the production of stem cells, which can be identified by the method according to this invention, was selected from the group consisting of interleukins and growth factors.
 More preferably, the interleukin this invention was selected from a group comprising interleukin 1 alpha, interleukin 1 family member 5 (Delta), interleukin 1 family member 6 (Epsilon), interleukin 1 family member 7 (Zeta), interleukin 1 family member 8 (ETA), interleukin 1 family member 9, interleukin 1 family member 10 (theta), interleukin 2, interleukin 3, interleukin 4, interleukin 6, interleukin 8 and interleukin 18.
 More preferably, the growth factor for this invention was selected from a group including CSF, GM-CSF, VEGF, EGF, FGF, IGF and PDGF.
 the compositions of this invention suitable for regulating the condition of skin, visible and/or tangible inhomogeneities of the skin (especially on the surface of the skin; such heterogeneity is usually undesirable). Such heterogeneity can the be induced or caused by internal and/or external factors and include signs of skin aging, described in this application. The term "regulating skin condition" includes preventive regulation and/or therapeutic regulating skin condition, including visible and/or tangible heterogeneity in the skin. In the context of this application preventive regulating skin condition includes delay, minimizing and/or preventing visible and/or tangible inhomogeneities in the skin. In the context of this application therapeutic regulation of skin condition involves reducing the intensity of symptoms, i.e. reducing, minimizing and/or remove inhomogeneities in the skin. The regulating skin condition includes improvement of skin appearance and/or tactile sensations. In addition, the compositions of this invention are particularly useful in the treatment of acne or other skin inflammations.
 the compositions of this invention are particularly useful for regulating signs of skin aging, mainly highly visible and/or tangible discontinuities in the texture of the skin associated with aging. "Regulation of signs of skin aging includes preventive regulation and/or therapeutic regulation of one or more of these signs (similarly, the regulation of this sign of aging, i.e. wrinkles, wrinkles or pores, includes preventive regulation and/or therapeu the practical regulation of this topic). In the context of this application of preventive regulation of this trait includes the delay, minimizing and/or preventing signs of aging. In the context of this application therapeutic regulation of such evidence involves reducing the intensity of symptoms, i.e. reducing, minimizing and/or removing signs of aging.
 the Signs of aging skin include, without limitation, all symptoms that are visible to the eye or detectable by touch, as well as all other macro - or microeffects caused by skin aging. Such signs may be induced or caused by internal factors or external factors, i.e. age ageing and/or the influence of environmental conditions. These signs may result from processes which include, inter alia, the development of inhomogeneities textures, such as wrinkles, including fine surface wrinkles, and large deep wrinkles, skin folds, fractures, tumors, large pores (i.e., associated with accessory structures, such as ducts of sweat glands, sebaceous glands, or hair follicles), flaking and dandruff and/or other forms of unevenness or roughness, loss of skin elasticity (loss and/or inactivation of functional elastin in the skin), sagging (including puffiness in the eye area and cheeks), the loss of platnost the skin, the loss of the tightening of the skin, loss of skin return after deformation, discoloration (including circles), spotting, discoloration, skin with hyperpigmentation, such as aging spots or freckles, corneal tumors, pathological differentiation, hyperkeratinization, elastosis, collagen breakdown, and other histological changes in the stratum corneum, dermis, epidermis, vascular system of the skin (i.e. telangiectasia or spider veins), and underlying tissues, especially those located close to the skin.
 the Percentage of the drug in this invention is from 0.0000001%to 10%, preferably from 0.000001 to 0,00001% by weight of the composition.
 the compositions of this invention typically are prepared so that the pH was about 4.5 and about to 9.5, more preferably, he was about 5 and about 7.5.
 For proofs of the invention are the following examples. Examples are given only for explanation and are in no way limit the scope of the invention.
 This example demonstrates the identification methods of preparation, suitable to stimulate production of stem cells in the skin.
Males white mouse BALB/C mice aged 5 days randomly divided into experimental group (group size n=10) and control groups is (n=10). All the experiments were carried out in accordance with the Principles of the observations of laboratory animals, formulated by the NIH. The cream containing the test drug applied to the skin of mice in the experimental group once a day for 5 weeks. Mice of the control group received the drug environment. At the end of the experiment, samples of the skin exposed to the drug were obtained from mice under anesthesia. Was made immunohistochemical analysis of skin samples for marker stem cell levels OST using goat polyclonal antibodies OST (laboratory Santa Cruz Biotechnology, Santa Cruz, California in a solution of 1:8000). Briefly, skin samples were fixed in 10% formalin and then paraffin according to the standard procedure. Paraffin sections were cut at 4 µm and dewaxed. Endogenous peroxidases were blocked with 3% hydrogen peroxide. Then the sections were heated for 5 min in 10 mm citrate buffer, to destroy nucleomegaly, were treated with 1% BSA at room temperature for 40 min and were placed in a thermostat at night at 4°C with primary antibodies OST. The next day, the sections were incubated (step 1) with biotinylated anti-goat immunoglobulin (laboratory Santa Cruz Biotechnology, Santa Cruz, CA)used at 1:200 solution and (step 2) avidin-biotinyl complex, conyuge ofanim with peroxidase from horseradish. Was performed staining with 3-amino-ethyl-carbazole and hydrogen peroxide. And in the end a contrast staining with Mayer hematoxylin. Painted by immunohistochemical image slices of the skin was examined under an optical microscope. Estimated number OST-positive cells / mm2. The results are expressed in crednerite the density increases OST-positive cells compared with the control group. The difference between groups was considered significant when p<0.05 (unidirectional ANOVA).
Identification of a drug candidate. The purpose of the testing was to identify drug candidate suitable to stimulate production of stem cells in the skin. We used five drug candidates. The cream containing the test drug or the environment, was made in the usual way. The cream was applied to the skin of mice, as described in method. The density of stem cells in the skin was evaluated by immunohistochemical method, as described above. When processing four test drugs no effect on the production of stem cells was not detected. One test drug, interleukin 1 alpha, showed a significant and 1.9-fold (p<0.05) increase in the level of stem cells in the skin compared with the control group. Thus, interleukin 1 alpha was identified as a drug candidate, brightneedle stimulate production of stem cells in the skin.
 This example shows the composition to stimulate production of stem cells in the skin
The percentage of ingredients, in weight%
Interleukin 1 alpha 0,000001, Citrate buffer as necessary to obtain a pH of 5.5, distilled water to 100
Manufacturing solution: the ingredients are mixed in the usual manner for the manufacture of the composition. Way to stimulate the production of stem cells in the skin: 1 ml of the composition is locally applied to the scalp, leave it on the skin for a period of time not less than about 15 minutes.
Composition to stimulate production of stem cells in the skin, including interleukin 1 alpha and dermatologically-acceptable diluent or carrier.
SUBSTANCE: method involves dissolving 855 mg of a crystalline hydrate of copper chloride (CuCl2·2H2O) in 100 ml of distilled water (concentration of Cu2+ ions in the prepared solution is 50 mmol/l) and adding 1 ml of the prepared solution to 100 ml of a standard reagent used in glucose oxidase test. The ascorbic acid oxidant used is copper chloride solution in end concentration in the glucose oxidase reagent of 500 mcmol/l.
EFFECT: method enables correct determination of glucose content.
1 tbl, 1 ex
SUBSTANCE: workers' blood serum is analysed for interleukin 4, protein S-100β, protein S-100 autoantibodies, voltage-dependent Ca-channel autoantibodies, glutamate receptor autoantibodies, γ-aminobutyrate receptor autoantibodies, dopamine receptor autoantibodies; diagnostic coefficients F1 and F2 are calculated; if the value F1 is less than F2, the early changes of the nervous system are diagnosed for the chronic exposure to vinyl chloride; F1 more or equal to F2 enables stating the absence of any signs of the chronic exposure to vinyl chloride. The developed method may be used in the periodic medical screenings, medical examinations of workers to diagnose some occupational diseases.
EFFECT: use of the invention improves higher accuracy of identifying the various signs of the chronic exposure to vinyl chloride through the use of a complex of the immunological structures of nerve tissue in the chronic exposure to vinyl chloride.
1 tbl, 2 ex
SUBSTANCE: invention may be used to predict a developing myocardial dysfunction in the children with acute lymphoblastic leukemia (ALL) at different stages of polychemotherapy (PCT). The method involves the blood examination for the iron metabolism parameters, namely before the beginning of polychemotherapy (1) and after the induction of remission (2), blood serum ferritin, hepcidin and iron are evaluated in the patients; the derived values are inserted into the equations to calculate varying ECG, IMS, B(E-Ea) NT-pro-BNP after the completion of the intensive PCT course (3) and the total coefficient K is calculated by formula K=ECG3* IMS3* B(E-Ea)3* NT-pro-BNP3, wherein a probability of the myocardial dysfunction is stated by the total coefficient, namely: the coefficient K> 0.24 ensures predicting the developing cardiac complications, while K <0.24 show a lower risk of the cardiac complications.
EFFECT: possibility to detect a risk of the developing myocardial dysfunction accompanying the early PCT by the biochemical parameters, namely in terms of iron metabolism.
1 tbl, 1 ex
SUBSTANCE: method consists in determining a characteristic profile of a test sample of a human biological fluid. It is concentrated off-line. Biologically active substances are separated using complexing additives, and a 'reference' is determined. Steroid hormones are taken as the analysed biologically active substances. The hormones are separated by performed by reversed-phase HELC in gradient elution using a diode array detector. The steroid profiles are used to form a matrix of the analytical signal intensities and the retention factors of each steroid. Each sample is graphically imaged by method of principal components, and the graphical images are used to form 'reference' and deviation clusters. The 'reference' and deviation clusters are corrected by soft independent modelling of class analogy taken as a reference. The pathologies are diagnosed by an ability of the patient's image to come with a 'reference' or a deviation.
EFFECT: reliable diagnosis of the pathologies associated with adrenal cortical diseases.
6 dwg, 2 ex
SUBSTANCE: what is presented is a method for prediction the efficacy of the anti-TNF therapy in the patients with rheumatoid arthritis on the basis of genetic typing the polymorphisms of TNF-alpha proinflammatory cytokine. The allelic polymorphism of the TNF-alpha gene promoter is studied in position 857. If the heterozygous state (genotype - 857ST) or the homozygous T allele carriers (genotype - 857TT) is identified, a high probability of the successful infliximab therapy is predicted. If identifying the homozygous allele C carrier in position - 857 of the TNF-alpha gene promoter (genotype - 857SS), a high probability of the failed infliximab therapy is predicted.
EFFECT: invention enables the rapid and effective prediction of the clinical outcome of the anti-TNF therapy in the patients with rheumatoid arthritis by one polymorph position.
2 tbl, 1 ex
SUBSTANCE: menopausal women with an endometrial hyperplastic type having complaints about spotting undergo biopsy of the lining of the uterus to determine endometrial progesterone and testosterone, if observing progesterone falling within the range of 2.0 to 7.0 ng/g of tissue and testosterone falling within the range of 4.0 to 8.8 ng/g of tissue, developing endometrial cancer is predicted, while progesterone within 24.0 to 29.6 ng/g of tissue and testosterone within 16.8 to 22.4 ng/g of tissue enable predicting developing uterine fibroid. The technical and economic effectiveness of the method consists in the fact that the detected levels of progesterone and testosterone in the intact endometrial tissue in the menopausal patients with a hyperplastic type are high-information laboratory indicators of the presence of either malignant, or benign uterine pathology, which can be used to form the groups of patients with the high risk of malignancy in the body of the uterus.
EFFECT: method is available, quick to implement.
1 tbl, 2 ex
SUBSTANCE: laboratory diagnostic technique for the small-dose poisoning with organophosphorous agents consists in assessing catalytic activity of blood plasma aryl esterase heated to 55°C for 5 min with indophenyl acetate used as a substrate. Catalytic activity of the enzyme and its increase after heating are assessed once after a contact with a organophosphorus agent; the effect of increasing catalytic activity of blood plasma aryl esterase after heating is expressed in %, taking catalyst activity of blood plasma aryl esterase before heating as 100%. In case activity of plasma aryl esterase after heating is increased by more than 20%, the small-dose poisoning with organophosphorus agents is diagnosed.
EFFECT: use of the declared technique enables stating effectively the fact of the small-dose poisoning with OFAs in the absence of any clinical signs of poisoning.
1 tbl, 2 ex
SUBSTANCE: method for prediction the recurrence-free survival period in cervical cancer (CC) is implemented by blood plasma catalase activity test, and the local process of cervical cancer (FIGO stage Ib-IIa) with activity 0.041 to 0.113 mmol/min/l in the patients enables predicting 50% probability of the 18-month period of the recurrence-free survival period, while blood plasma catalase activity 0.008 to 0.035 mmol/min/l shows 80% probability of the recurrence-free survival period.
EFFECT: prediction of the recurrence-free survival period in local cervical cancer.
SUBSTANCE: patient with advanced cervical cancer (FIGO stage IIb-IV) is examined for tumor tissue glutathione reductase activity. When tumour tissue glutathione reductase activity is 10.9 to 14.9 mmol/min/mg of protein, the patient is expected to have 18% probability of the 18-month recurrence-free survival period, while tumour tissue glutathione reductase activity being 15.2 to 18.5 mmol/min/mg of protein shows 60% probability of the 18-month recurrence-free survival period.
EFFECT: use of the declared method enables predicting the recurrence-free survival period in local cervical cancer.
SUBSTANCE: there are presented methods for prediction of a higher risk of hypertension related to the VEGF antagonist therapy in a patient. What is involves is screening of a sample recovered from the patient for the purpose of VEGF (-1498C/T) and VEGF (-634G/C) genome polymorphisms. If the genotype comprises the VEGF (-1498C) and/or VEGF (-634G) polymorphisms, the patient is likely to have a higher risk of hypertension related to the VEGF antagonist therapy. What is presented is a kit for prediction of a higher risk of hypertension in the patient, related to the VEGF antagonist therapy, comprising a first oligonucleotide and a second oligonucleotide specific for polymorphism in the VEGF gene selected from the group consisting of VEGF (-1498C/T) and VEGF (-634G/C).
EFFECT: effective methods and kits for prediction of a higher risk of hypertension related to the VEGF antagonist therapy in the patient.
12 cl, 5 tbl, 1 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: compounds of the present invention are novel peptide analogues of oxyntomodulin (oxm) wherein one or more amino acid residue of the sequence oxm are replaced. The replacement of amino acid residues 15-24 of the peptide oxm either by amino acid residues 968-977 of the α-latroxin peptide (and versions thereof), or by amino acid residues 15-24 of extendin-4 (and versions thereof), or combining the amino acid residues of these sources, and/or the replacement of amino acid residues 27-33 of the peptide oxm by amino acid residues 27-33 of extendin-4, and/or adding the amino acid residues to an C-terminal of the peptide, enables producing a number of the analogues oxm presenting oxm-like activity to reduce food consumption, and according to some other aspects, more evident ability to reduce food consumption.
EFFECT: reduced food consumption.
32 cl, 779 dwg, 106 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: group of inventions refers to biochemistry. What is presented is pegylated interferon-α2b (IFN-α2b) having a structure as shown in the patent claim. Pegylated interferon-α2b is prepared by combining IFN-α2b with branched Y-polyethylene glycol (YPEG), wherein YPEG is bound to IFN-α2b by an amide bond formed by ε-amino group of a side of Lys residue in IFN-α2b in the position of 134 in SEQ ID No. 1. A method for preparing and purifying pegylated IFN-α2b involves the stages as follows. In an alkaline medium at pH 9.0, branched Y-polyethylene glycol is reacted with IFN-α2b to prepare pegylated IFN-α2b. The prepared reaction products are recovered by an anion exchange resin, and the given products are eluted by an anion gradient to prepare modified products. The anion exchange resin is Q Sepharose FF, and the anion gradient is a chloride ion gradient. Further, the modified products are eluted by a cation exchange resin with a cation gradient. The cation exchange resin is SP Sepharose FF, and the cation gradient is a sodium ion gradient. Thereafter, each peak is collected separately. The product activity of each peak is determined to choose a peak corresponding to the reaction product having the highest activity. What is also presented is a formulation for treating a disease requiring IFN-α2b to be used, which consists of a pharmaceutically effective amount of said pegylated IFN-α2b and a pharmaceutically acceptable carrier, or an inactive substance. Pegylated IFN-α2b or the above formulation is also used for preparing a medicine for treating various diseases requiring IFN-α2b to be used.
EFFECT: presented pegylated IFN-α2b has a higher specific activity of 2,65±0,185×106 IU/mg and a prolonged serum half-life.
26 cl, 11 dwg, 5 tbl, 3 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: group of inventions refers to medicine. The invention aims at creating biodegraded implants of the regenerated silk fibroin Bombyx mori. A method for preparing the biodegraded composite matrix of the regenerated silk fibroin Bombyx mori involves the stages: - dissolving the regenerated fibroin in a buffer containing CaCl2, C2H5OH, H2O in molar ratio 1:2:8. That is followed by dialysis of the prepared solution to the fibroin concentration min. 20 mg/ml. The fibroin solution of the concentration min. 20 mg/ml is mixed with dimethyl sulphoxide (DMSO). Gelatin or polylysine is added to the prepared mixture. The prepared mixture is frozen and thawed with an organic solvent added; what is also presented is the use of the specified biodegraded composite matrix as a base for the biodegraded implant.
EFFECT: group of inventions enables improving the biocompatibility, increasing the adhesive properties of matrix and cell proliferation in a mammalian body.
2 cl, 3 dwg, 3 ex, 1 tbl
FIELD: medicine, pharmaceutics.
SUBSTANCE: group of inventions refers to medicine, namely biopharmaceutics, and may be used for making vaccines. For this purpose, the vaccine containing the influenza VLPs comprises the Ml, HA and NA proteins of the influenza virus wherein said vaccine induces a long-lived immunity to influenza virus infection in an individual susceptible to influenza, and wherein the MI protein of influenza is prepared of the influenza strain A/Indonesia/5/05. The group of inventions also refers to the use of the vaccine of the composition mention above for the induction of the long-lived immunity to influenza virus infection in an individual.
EFFECT: use of the given composition of the vaccine enables the most effective method to prepare an amount of VLPs necessary for the vaccine production.
21 cl, 41 dwg, 31 ex
SUBSTANCE: in order to reduce content of dimers in a composition which contains a factor VII polypeptide or a version of factor VII polypeptide, pH of said composition is brought to a value ranging from 5 to 10.0; the composition is heated to temperature ranging from 20°C to 50°C for 10 minutes to 72 hours, followed by cooling to temperature not higher than 5°C.
EFFECT: low dimer content in a factor VII polypeptide composition by heat treatment.
8 cl, 4 dwg, 3 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: invention refers to biotechnology and virology. Any two E2 encoding nucleic acid molecules have the identity percentage less than 75% in any part of 40 or more unbroken nucleotides. There are also described an infectious viral particle and a composition containing the mentioned expression vector.
EFFECT: what is disclosed is the expression vector containing the nucleic acid molecules coding at least two papillomavirus E2 polypeptides.
34 cl, 3 dwg, 1 tbl, 11 ex
SUBSTANCE: what is presented is a method for differentiation of osteoarthritis, rheumatoid arthritis, and non-pathological states in a sample, including the measurement of the concentration of the peptide of a C1 protein 2 chain of the cartilage intermediate layer (CILP-2). The high level of the C1 protein 2 chain CILP-2 compared with rheumatoid arthritis and the normal state indicates that the patient has osteoarthritis. What is also presented is a peptide for differentiation of osteoarthritis and rheumatoid arthritis and non-pathological states, comprising SEQ ID NO:1. There are presented antibodies immunoreactive to the mentioned peptide, and a kit containing the antibodies and the analysis regulation.
EFFECT: effective agents and methods for differentiation of osteoarthritis, rheumatoid arthritis and non-pathological states by the increase of the concentration of the C1 protein 2 chain of the cartilage intermediate layer in the sample.
7 cl, 2 dwg, 4 ex
SUBSTANCE: method includes the following stages: conservation of cells in presence of buffered 80-90% glycerine, collapse of cell shells by 3% triton X-100, extraction by increasing concentrations of salts: 0.14 M, 0.35 M; 2 M NaCl, 6 M guanidine hydrochloride with 0.1% β-mercaptoethanol, extraction of positively charged proteins from above fractions with the help of ion-exchange chromatography with amberlite IRC-50 in an interrupted gradient of guanidine hydrochloride: 6%, 8.9%, 10.6%, 13% on 0.1 M potassium-phosphate buffer pH 6.8 and detection of sites of sensitivity in them to Arg-X proteolysis.
EFFECT: invention may be used in analysis of molecular-genetic mechanisms of procaryote cell structure formation and role of protein components in their organisation, and also when studying features of genome remodelling, which is necessary for opening of paths of regulation mechanisms of macro- and microorganisms.
9 dwg, 1 ex
SUBSTANCE: methods are described to prevent a virus flu infection and detection of efficiency of a flu virus vaccine using a molecule of hemagglutinin (HA) of a flu virus A of subtype H5, immunogenicity of which is increased by replacement of amino acids in the HA sequence.
EFFECT: replacement of specific remains in HA, such as introduction of asparagin into the position 223 in HA H5, makes it possible to increase sensitivity of hemagglutination moderation reaction as a result of variation of receptor specificity or ability to binding of antibody-antigen.
24 cl, 7 tbl, 7 ex
SUBSTANCE: invention discloses artificial peptide mini-antigens (PMA) which can be used to induce controlled protective humoral IgG-mediated immune response against allergen. The mini-antigen includes: hydrophobic peptides from different proteins and any pathogens which are linked with pockets of the most common MHC (major histocompatibility complex) molecules class II and to which, in the human population, there are memory T cells (memory T cell T epitopes (MTC)), and hydrophilic peptides from different proteins which are on the surface of wild type allergen, which are conjugated onto the support of the cladding of the polymer particle and are accessible for antibodies. (The structure of the mini-antigen is shown on fig 14 in the description). The mini-antigens are characterised by the capacity to replace B epitopes with peptides of another allergen while preserving the nucleus of the MTC T epitopes, which enables to use the peptide mini-antigens to make anti-allergy vaccines against different types of allergies. The presence of memory T cells helps to considerably speed up immune response triggering. The introduction of peptides of only basic allergen proteins into the structure ensures anti-allergy protection with minimal load on the immune system.
EFFECT: absence of binding of IgE with peptides makes vaccination safe and fast.
14 dwg, 2 tbl, 3 ex
FIELD: medicine, pharmaceutics.
SUBSTANCE: group of inventions relates to water composition, which includes system with high content of surface-active substances and to method of claimed system obtaining. Claimed composition is transparent and includes system, which contains microfibrous cellulose in concentration from 0.05% to 0.15% wt/wt, surface-active substance in concentration from 51% to 99% wt/wt, and suspension particles of inclusions. Method of obtaining system with high content of surface-active substances includes combination of microfibrous cellulose with water and mixing with high shear strain, addition of surface-active substance and following mixing and addition of inclusion particles while mixing.
EFFECT: group of inventions ensures obtaining stable and transparent compositions with high content of surface-active substances.
20 cl, 4 ex