(by Johann Wolfgang von Goethe)
А new technique of mammalian bone marrow cells culturing with "duplex" decontamination system has been developed. This technique is intended for generating of progenitor cells with high plasticity. The subject matter of the declared technique of progenitor cells culturing is creation of favourable aseptic and growth conditions in presence of cytokinesis regulators. The partial culture medium regeneration by depletable components has been performed while growing of cell monolayer. The present technique refers to the first animal cells and tissues culture system (monolayer systems where cell proliferation is taking place on the surface of immobilized supports submerged into culture medium). The novelty of the proposed technique is the duplex decontamination practice, where agar-agar carrier was diffusely saturated with Argentum ions (Ag2+) in combination with natural polyphenol compounds, and thiazol-containing compounds of fluconazole type have being added during culture solution formation (ЕР 5.0; 5.6, р. 4581). The 2MEM standard culture medium (Adams R.L.P., 1990) was used as culture solution base. Addition of hormonal growth compounds (interleukins, insulin, gonadotropin and others) together with cytokinesis regulators during cell culture growth process enabled the performing of long-lasting cultivation without any passage, and generating of the relatively homogeneous culture of progenitor cells with rather high vital potential. The experimental studies of generated progenitor cells culture were tested in pubertal BALB/c mouse males with weight of 18-21 g, being fed with cubed feed in standard vivarium conditions. The experimental animals were injected intraperitoneally by 0.18-0.22 ml of the cell suspension in aseptic conditions. The microscopy of blood smears, performed in 48 hours after the beginning of the experiment, demonstrated the cluster
generation likely from young forms of stem cells surrounded by the formed elements. In this particular case, the endogenic microcarriers can be represented as different spherical organic blood aggregates carrying the required adhesive surface-bound charge. The results obtained from blood smear microscopy enable assuming the optimum size of endogenic microcarriers – from 30 up to 70 μm. Considering the cytometric sizes of progenitor cells [10-12 μm] (Van Bekkum D.W. и соавторы, 1971; Van Bekkum D.W., 1990), it can be suggested that the part of cells settles down on the endogenic microcarriers during their migration through the rich vascularised peritoneum, and forms the active growth colonies of adherent monolayer. The presence of optimum conditions for cell growth and differentiation in blood supposes that the processes of active cell transformation take place directly in animal blood. The studying of this phenomenon is still going on.
Patient man Mr. Ivanov S.S. (born in 1938) with osteomyelofibrosis, was performed the control blood examination (March, 18, 2003) at the Pulmonological Department of Lubny City Hospital No7 before starting of anthocyanins administration (fractions SAM-7bc and SAM-7bb); According to the examination results, the total amount of leukocytes was 67.0×109/l, which was 6.7 times higher than normal. The Patient was diagnosed leukocytosis, granulocytosis with nuclear shift to the left, moderate anisocytosis and macrocytosis. From March 18, 2008, the Patient was suggested to start receiving of anthocyanin concentrate made from black currant and dewberry berries (fractions SAM-7bc и SAM-7bb) per os , at dose of 14.0 grams daily diluted in 1000.0 ml of slightly acidulouss mineral water along with keeping the fat-free diet. The control blood examination (March 28, 2008) showed the actual decreasing of leukocyte count up to 43.4×109/l, that was 4.34 times higher than upper normal limit with decreasing of multiplicity in 1.54 times regarding the baseline leukocyte count. In general, the patient’s condition was sufficient, and there was steady tendency for renewal of primary biological motivations, such as food motivation, appetite increasing and sleep improvement. The control blood examination (April 24, 2008) taken after 40 days since starting of receiving of anthocyanin concentrate, showed the actual decreasing of blood leukocytes amount up to 11.9×109/l, which yet was 1.19 times higher than the upper limit, accompanied with 5.6 times decreasing of multiplicity vs the initial leukocyte amount. Herewith, there were
also positive changes of relevant blood values, e.g. ESR decreased from 50 mm/h (clinical blood assay taken on March 18, 2008) to 15 mm/h. Later on the fluctuation of the blood leukocytes rate within the limits of 11×109/l - 21×109/l has been observed until the middle of May, 2008. Such history of blood leukocytes rate fluctuation was associated with the episodes of discontinuation and renewing of polyphenols intake. The Patient renewed the anthocyanins intake at dose from 12 to 14 grams per day with monthly interval of 2-5 days since May 15, 2008. The patient was keeping the dietary restrictions and have being on the fat-free diet. The control blood examination (December, 2008) showed the leukocytes rate 19.9 ×109/l, which was 1.19 times higher than normal limit along with Patients’ sufficient condition and restoration of working capacity . The Patient was in a cheerful mood and had active attitude to the milieu. Clinical blood assay dated January 14, 2009 confirmed the tendency for lasting stabilization of blood leukocytes rate that was 11.7×109/l, and was 1.17 times higher that the upper normal limit. The Patients’ condition is satisfactory, physiological status is stable. During the whole period of receiving of the drug, the Patient was not observed any digestive and hypersensitivity reactions to anthocyanin concentrate. The cancerous disease dynamics, given above, shows the beneficial potential of natural polyphenols, and requires the necessity of further scientific research of black currant and dewberry berries SAM-7bc and SAM-7bb fractions as effective alternative of traditional chemotherapy.
Five-year-old boy Dima with a diagnosis: bronchial asthma, mixed type, severity of middle, exacerbation phase. In laboratory conditions were detected the following changes, combining with enzyme systems activation: impairment of pseudoreactions of T-lymphocytes, B-cells and phagocytes dysfunction [analysis after date 20/08/2008: EA-rosette-forming cell:19%(N 32-41%); EA-rosette-forming cell abs: 0,38×109/l (N 1.07-2.0×109/l); CD3: 40% (46-63%); CD3 abs: 0.8×109/l (N 1.5-3.15×109/l); CD22: 11% (N 13-22%); CD22 abs: 0.22×109/l (N 0.4-1.1×109/l); CD4: 22% (N 35-43%); CD8: 16% (N 22-26%); immunoregulatory balance: 1.37 (N 1.1-2.2); IgA: 2.4g/l (N 0.52-2.2g/l); IgM: 1.5g/l (N 0.4-2.0g/l); IgG: 11.0g/l (5.4-14.2g/l); IgE: 580.0 IU/ml (N up to 60.0 IU/ml); Leukocytes: 5.4×109/l (N 4.0-9.0×109/l); Lymphocytes: 37% (N not less 45%)].
On the grounds of factors changes of immune status we considered these alterations as immunodeficiency and, on these grounds, the child was recommended to
take immunostimulating drugs on the basis of polyphenol compounds extracts, the effect of which is concentrated on the correction of detected disturbances in immune system.
As a result of the therapy carried out by new drugs on the basis of polyphenol compounds extracts, considerable progress in immune status factor was noted as well as acute phase was stopped and disease severity was reduced [analysis after date 23/12/2008: EA-rosette-forming cell:40%(N 32-41%); EA-rosette-forming cell abs: 1,94×109/l (N 1.07-2.0×109/l); CD3: 44% (46-63%); CD3 abs: 2.1×109/l (N 1.5-3.15×109/l); CD22: 24% (N 13-22%); CD22 abs: 1.16×109/l (N 0.4-1.1×109/l); CD4: 22% (N 35-43%); CD8: 21% (N 22-26%); immunoregulatory balance: 1.04 (N 1.1-2.2); IgA: 1.45g/l (N 0.52-2.2g/l); IgM: 1.1g/l (N 0.4-2.0g/l); IgG: 11.9g/l (5.4-14.2g/l); IgE: 537.0 IU/ml (N up to 60.0 IU/ml); Leukocytes: 9.0×109/l (N 4.0-9.0×109/l); Lymphocytes: 54% (N not less 45%)]. Therapeutic measures and supervisions of patient are in progress.
Sick man Djafarov V.S., born in 1952, had an inpatient treatment from January 2008 with a diagnosis: agnogenic exudative pleurisy. In past history were complains about: evident dyspnea; weakness; usually bad short cough, productive cough with a discharge of sputum often in the mornings; weight and appetite reduction; chest pains; temperature rise up to subfebrile figures (no higher than 38 C); weight loss. A standard antibacterial and antifungal therapy with a limited improvement was carried out in steady-state conditions.
In July, 2008 after discharge from the clinic the patient began an intake of polyphenols extract, 30-50 drops
daily without changing a habitual food ration. In two months general condition of the patient has considerably improved: dyspnea, weakness and chest pains have vanished, productive cough with easily discharging sputum has noted occasionally, an appetite has improved, the appearance of an optimistic mood and a cheerfulness with a lasting tendency to positive changes in the intellectual-amnestic sphere has noted. By December, 2008 the patient's cough was completely stopped and concomitant symptomatology was cut short. Supervision of the patient is in progress.
In December 3, 2008 Nikolay V. Ryzhkov, born in 1947, the resident of Zmievskiy district (Kharkov region), was hospitalized with complains about weakness; hyperhidrosis; cough with an elimination of a bulk of dense purulent sputum; dyspnea; temperature rise up to 39ºC; backaches. A rounded shadow about 3mm diameter with indistinct contour on the left in the third intercostal space was objectively noticed on the X-ray film of December 5, 2008, as well as calcified paratracheal lymph node on the left. Thus, in December 24, 2008 the patient was diagnosed the lungs lesion: Focal lesion of the right lung. Hamartoma or progonoma (nodular tumourous mass). A diagnostic bronchoscopy has shown the absence of atypical cells. Leucocytes in the whole field of view; erythrocytes, invariable in some areas; some epithelial and alveolar cells were noticed; atypical cells and Koch bacillus (BK) weren’t found in the sputum analysis of December 5, 2008. Leucocytes – 9,0×109/l, eosinophils – 1%, stab neutrophils – 10%, segmented neutrophils – 53%, lymphocytes – 33%, monocytes – 2%, ESR – 20 mm/h were noticed in the clinical blood analysis of December 5, 2008. Cholesterin – 6,2 millimole/l; bilirubin – 18,7 millimole/l; urea – 6,7 millimole/l; creatinine – 88,0 micromole/l were noticed in the biochemical blood analysis. During a week antibacterial therapy was carried out. Upon its completion laboratory reanalyses – sputum analysis and clinical blood analysis – were carried out. Leucocytes ¼ of the field of view; invariable solitary erythrocytes, some epithelial and alveolar cells were noticed, atypical cells and Koch bacillus (BK) weren’t found in the sputum analysis of December 12, 2008. Leucocytes – 6,1×109/l, eosinophils – 1%, stab neutro-
phils – 4%, segmented neutrophils – 58%, lymphocytes – 34%, monocytes – 3%, ESR – 13 mm/h were noticed in the clinical blood analysis of December 15, 2008. The rounded indistinct shadow remained in a repeated film. Tumour marker analyses of January 9, 2009 have shown the following: CEA (carcinoembryonic antigen) – 49,9 (the norm is up to 5,0); Alfa-Fetoprotein analyses – 2,3 (the norm is up to 10,0); prostate-specific antigen – 7,0 (the norm is up to 10,0). From January 8, 2009 a conventional management of the patient, using celandine tincture, breast tea and Dorogov’s ASD F-2 solution, has begun. Simultaneously with the therapy carried out from January 8, 2009, the conservative therapy was amplified with the intake of polyphenol compounds per os during 10 days. The reanalysis of sputum of March 25, 2009 has shown the following: leucocytes ½ of the field of view; erythrocytes are absent; solitary epithelial and alveolar cells; Koch bacillus (BK) weren’t found. Leucocytes – 5,2×109/l, eosinophils – 1%, stab neutrophils – 4%, segmented neutrophils – 61%, lymphocytes – 32%, monocytes – 2%, ESR – 9 mm/h were noticed in the clinical blood reanalysis of March 25, 2009. Cholesterin – 5,9 millimole/l; bilirubin – 17,2 millimole/l; urea – 6,1 millimole/l; creatinine – 77,0 micromole/l were noticed in the biochemical blood analysis of March 26, 2009. Tumour marker analyses of March 18, 2009 have shown the following: CEA (carcinoembryonic antigen) – 16,3 (the norm is up to 5,0); Alfa-Fetoprotein analyses – 2,0 (the norm is up to 10,0); prostate-specific antigen – 0,71 (the norm is up to 10,0). Currently the patient’s general condition is stable and satisfactory. Cough, dyspnea and sputum aren’t noticed.
...the part of cells settles down on the endogenic microcarriers and forms the active growth colonies of adherent monolayer... more...
...in some weeks after beginning of the reception of polyphenols the number of leukocytes in the blood was decreased in five times under the stabilization of other blood parameters... more...
![[Valid RSS]](images/valid-rss.gif "Validate my RSS feed")