Effects of reprogrammed splenic CD8+ T-cells in vitro and in mice with spontaneous metastatic Lewis lung carcinoma.

BACKGROUND: Metastatic disease is a major and difficult-to-treat complication of lung cancer. Considering insufficient effectiveness of existing therapies and taking into account the current problem of lung cancer chemoresistance, it is necessary to continue the development of new treatments. METHODS: Previously, we have demonstrated the antitumor effects of reprogrammed CD8+ T-cells (rCD8+ T-cells) from the spleen in mice with orthotopic lung carcinoma. Reprogramming was conducted by inhibiting the MAPK/ERK signalling pathway through MEKi and the immune checkpoint PD-1/PD-L1. Concurrently, CD8+ T-cells were trained in Lewis lung carcinoma (LLC) cells. We suggested that rCD8+ T-cells isolated from the spleen might impede the development of metastatic disease. RESULTS: The present study has indicated that the reprogramming procedure enhances the survival and cytotoxicity of splenic CD8+ T-cells in LLC culture. In an LLC model of spontaneous metastasis, splenic rCD8 + T-cell therapy augmented the numbers of CD8+ T-cells and CD4+ T-cells in the lungs of mice. These changes can account for the partial reduction of tumors in the lungs and the mitigation of metastatic activity. CONCLUSIONS: Our proposed reprogramming method enhances the antitumor activity of CD8+ T-cells isolated from the spleen and could be valuable in formulating an approach to treating metastatic disease in patients with lung cancer.

Age-Related Features of the Response of Cancer Stem Cells and T Cells in Experimental Lung Cancer.

The responses of tumor stem cells and various populations of CD4 and CD8 T cells of young and aged C57BL/6 mice were studied in a lung cancer model. Using Lewis lung carcinoma cell line, an orthotopic model of lung cancer was modeled. Cancer stem cells, circulating tumor cells, and various populations of CD4 and CD8 T cells in the blood and lung tissue were studied by cytometry. We revealed age-related differences in the content of various populations of CD4 and CD8 T cells in the blood and lungs of intact young and aged mice. Age-related features of the reaction of various populations of cancer stem cells and CD4 and CD8 T cells in the blood and lungs of animals in the Lewis lung carcinoma were shown.

The Role of Cancer and Somatic Stem Cells in the Anti-Inflammatory and Antitumor Effects of Aconitum baicalense Extract on Experimental Breast Cancer.

We studied the effects of the extract of the terrestrial part of Aconitum baicalense in BALB/c female mice at the early stages after the injection of N-methyl-N-nitrosourea (MNU). The extract reduced inflammatory activity and tumor growth in the mammary gland. The antitumor and anti-inflammatory effects of the extract are based on the inhibition of cancer stem cells, hematopoietic stem cells, and hematopoietic progenitor cells that promote inflammation. The extract of A. baicalense disrupted the recruitment of epithelial progenitor cells and angiogenesis precursors to the mammary gland preventing neovascularization and transformation of epithelial cells into tumor cells.

Age-Related Features of the Early Period of Liver Regeneration after Partial Hepatectomy in Rats.

Regenerative processes in the liver were studied in 3-month-old (young) and 9-month-old (aged) male Wistar rats on day 1 after 30 and 70% hepatectomy. Regardless of the resected liver volume, shifts in the biochemical parameters of the serum in aged rats were more pronounced than in young animals. After 30% hepatectomy, no age differences in the rate of hepatic regeneration were found, while after 70% liver resection this parameter was higher in young rats. Hepatectomy in young rats led to recruitment of MSC, hepatocyte precursors, endothelial and epithelial progenitor cells into the liver parenchyma and increased fluidity of the plasma and mitochondrial membranes of hepatocytes. In aged rats, the recruitment of MSC, hepatocyte precursors, and endothelial progenitor cells into the injured liver was impaired and the rigidity of the mitochondrial membranes of hepatocytes increased.

Early Diagnostic Markers and Therapeutic Targets for Experimental Breast Cancer.

Various stem cells were studied in female BALB/c mice at the early terms after administration of N-methyl-N-nitrosourea to search early diagnostic markers and therapeutic targets. At these terms, damage to the epithelium and endothelium, inflammation, and fibrosis were observed in the mammary gland, but the tumor was not detected. Cancer stem cells, hematopoietic stem cells (HSC), hematopoietic progenitor cells, angiogenic precursors, and epithelial progenitor cells were found in the blood and mammary gland. Cancer stem cells (CD44+CD24-) are proposed as the early diagnostic marker of breast cancer, and short-living HSC, hematopoietic progenitor cells, and angiogenic precursors (CD45-CD117+FLK-1+) as predictors of the formation of tumor microenvironment.

Age-Related Features of the Viscosity of Plasma and Mitochondrial Membranes of Hepatocytes in Liver Cirrhosis.

The viscosity of plasma and mitochondrial membranes of hepatocytes was studied in young (3-month-old) and old (9-month-old) male Wistar rats. It was shown that viscosity of hepatocyte plasma and mitochondrial membranes in young rats under optimal vital functions in the area of protein-lipid membrane contacts was significantly lower than in old rats. No age-related differences in the viscosity of lipid-lipid membrane contacts and in the polarity of protein-lipid contacts and lipid layers were found. Liver cirrhosis induced by carbon tetrachloride and ethanol administration was associated with increased fluidity of the plasma and mitochondrial membranes of hepatocytes in rats of both age groups. The decrease in membrane viscosity in young rats occurred due to a decrease of the viscosity in the area of protein-lipid and lipid-lipid contacts, while in old rats in the area of protein-lipid contacts. Carbon tetrachloride and ethanol did not affect the polarity of lipid contacts and lipid layers.

Age-Related Features of the Response of the Liver and Stem Cells during Modeling of Liver Cirrhosis.

We studied the age-related characteristics of the response of stem cells and liver in male Wistar rats to administration of carbon tetrachloride (CCl4) and ethanol. It was shown that modeling of liver cirrhosis caused inflammation, fibrosis, damage to sinusoidal capillaries, necrosis, and disturbances in the functional activity of hepatocytes in young rats. These processes were accompanied by mobilization of profibrotic mesenchymal stem cells (MSC), proinflammatory hematopoietic stem cells (HSC) and lymphocytes (CD45hiCD133+) from the bone marrow into the blood and migration to the liver. On the other hand, the number of hepatocyte precursors expressing Sox9 (cells of Hering's canal), immature cholangiocytes, Ito cells, oval cells, and endothelial cells of the liver sinusoids) sharply increased in the liver. In young rats, mobilization and migration of MSC, HSC, and hepatocyte precursors against the background of liver cirrhosis were more intensive than in old animals. The higher resistance of old rats to exposure is associated with age-related changes in the niches as well as in mobilization and migration of cells.

Genetic Factors as the Basis of Sex Differences in Damage to Lung Endothelium and Regulation of Angiogenesis Cells in Modeling Pulmonary Emphysema in C57BL/6 Mice with Dyslipidemia and Hyperglycemia.

We studied the formation of injuries in lung endothelium and the response of angiogenesis cells during modeling of pulmonary emphysema in male and female C57BL/6 mice with metabolic disorders. Hemodynamic disturbances and reduction in the area of the microvasculature caused by combined pathology in male mice were more pronounced than in females. Mobilization and migration of angiogenic precursors were impaired in both male and female mice. In males, activity of recruiting endothelial progenitor cells, vascular smooth muscle cells, luminal cells of nascent vessels and pericytes into the lungs was additionally reduced. In females, accumulation of endothelial progenitor cells (CD45-CD31+CD34+), vascular smooth muscle cells, and pericytes in the lungs was observed, which indicated activation of endothelial regeneration. Sex differences in the reaction of the lung endothelium and angiogenesis cells can be explained by genetic factors of lipid and glucose metabolism.

Blockade of Dopamine D2 Receptors as a Novel Approach to Stimulation of Notch1+ Endothelial Progenitor Cells and Angiogenesis in C57BL/6 Mice with Pulmonary Emphysema Induced by Proteases and Deficiency of α1-Antitrypsin.

We studied the effects of spiperone, a selective blocker of dopamine D2 receptors, on the model of pulmonary emphysema provoked by administration of elastase and D-galactosamine hydrochloride to female C57BL/6 mice and characterized by activation of proteases in the lungs and systemic deficiency of its inhibitor α1-antitrypsin. In this model, spiperone prevented the development of inflammatory reaction and reduced the area of emphysematous expanded alveolar tissue. The expression of angiogenic marker CD31 in the lungs increased under these conditions. Regeneration of the damaged microvascular bed under the action of spiperone resulted from recruiting of Notch1+ endothelial progenitor cells (CD45-CD31+CD34+) into the lungs and blockade of the inhibitory effect of dopamine on phosphorylation of VEGF-2 receptors in endothelial cells of different maturity. In addition, spiperone produced a protective effect on hepatocytes and restored the production and secretion of α1-antitrypsin by these cells.

Endothelial Progenitor Cells and Notch-1 Signaling as Markers of Alveolar Endothelium Regeneration in Pulmonary Emphysema.

We studied the effects of elastase, cigarette smoke extract, D-galactosamine hydrochloride, and tyrosine kinase inhibitor SU5416 on endothelial progenitor cells and angiogenesis precursors, as well as on Notch-1 expression by immature endothelial cells. Simultaneously with pulmonary emphysema, different damaging factors with diverse mechanisms of action caused pathological changes in the microvascular network of the lungs and destroyed the alveolar endothelium in female C57Bl/6 mice. D-galactosamine hydrochloride disturbed mobilization of endothelial progenitor cells expressing VEGFR (CD45-CD309+) and angiogenesis progenitors (CD45-CD309+CD117+) and their migration into emphysema expanded lungs. Elastase inhibited VEGFR-expressing endothelial progenitor cells, while cigarette smoke extract inhibited cells with CD45-CD31+CD34+ phenotype. In pulmonary emphysema provoked by elastase or D-galactosamine hydrochloride, angiogenesis was provided by endothelial cells with CD45-CD31+CD34+ phenotype, whereas in emphysema modeled with SU5416 or cigarette smoke extract, it was provided by the endothelial VEGFR-expressing cells and mature CD31+ endothelial cells, respectively. Replenishment of immature endothelial cells damaged by elastase and SU5416 involved Notch-1+ angiogenesis precursors and Notch-1+ endothelial progenitor cells with VEGFR.

[Cryoglobulinemia in a case of multiple myeloma associated with viral hepatitis B].

Cryoglobulinemia is a biochemical disorder accompanying many infectious, systemic, and lymphoproliferative conditions. Cryoglobulins are proteins capable of reversible precipitation or gel formation at temperatures below 37 degrees C. There are 3 types of cryoglobulins depending on the type of immunoglobulins in their composition; hence, three types of cryoglobulinemia occurring in different diseases. Clinical manifestations of cryoglobulinemia are sometimes inapparent, various combinations of the symptoms of the disease frequently occur. An example of atypical acute cryoglobulinemic vasculitis in a patient with multiple myeloma and viral hepatitis B is presented. Methods of diagnostics of this disease and its treatment are described.

[The application of combined physiotherapeutic methods for the complex treatment of patients with dyscirculatory encephalopathy].

This article reports the results of applying basic pharmacotherapy (enalapril, cytoflavin) and its combination with physical factors (transcranial electrostimulation, combined application oftranscranial electrostimulation and low-frequency magnetic therapy) in the complex treatment of patients with stage I-II dyscirculatory encephalopathy. The study has demonstrated that the combined treatment with cytoflavin, enalapril, transcranial electrostimulation and low-frequency magnetic therapy produced the most pronounced therapeutic effect (82.5%), as confirmed by positive dynamics of clinical and functional parameters.

[The use of topiramate at patients with combined craniovertebral anomaly].

Twenty-eight patients with concomitant cranio-vertebral anomaly: Kimmerle anomaly and Chiari type I malformation were examined. A main reason for visiting a doctor was headache, dizziness and sleep disturbance. Topamax (topiramate) was used for stopping the symptoms. It was prescribed in dosage 25 mg in the evening to 16 patients and in dosage 25 mg in the first two weeks and then in dosage 50 mg in the evening for 60 days - to 12 patients. The use of Topamax led to the reduction and /or disappearance of headache, sleep normalization and improvement of cerebral bioelectric activity. The drug was well tolerated.

[High temperature initiates changes of Wolbachia ultrastructure in the ovaries and early embryos of Drosophila melanogaster].

Electron microscopic analysis of Drosophila melanogaster (w1118) ovarian cells has demonstrated that stressful heat treatment of flies results in the appearance of electron dense granules and large lysosomes in the cytoplasm of ovarian cells, which is not related with the presence of Wolbachia, as these changes are observed in both the infected and uninfected flies. High temperature initiates essential envelope defects and other structural changes of symbiotic bacteria in the cytoplasm of ovarian cells. Some embryos developing from eggs of heat shocked flies die, however, bacteria in the survival embryos retain their typical morphology. Endosymbionts do not change their localization and their contacts with the mitochondria and endoplasmic reticulum in the ovarian cells and early embryos after heat shock treatment of the flies. The results obtained show that high temperature influences on both the host and the endosymbiont, but does not change their structural mutual interactions.

[SMC-electrophoresis of peat mud in the treatment of patients with duodenal ulcer in outpatient care unit]. / Opyt lecheniia bol'nykh iazvennoi bolezn'iu dvenadtsatiperstnoi kishki CMT-élektroforezom torfianoi lechebnoi griazi v usloviiakh dnevnogo statsionara.

To study collagen metabolism in reparative regeneration of duodenal ulcer in response to peat mud electrophoresis of Uva sanatorium using sinusoidal modulated currents (SMC) as monotherapy in a day hospital, 82 duodenal ulcer patients in exacerbation or attenuation were examined. They received SMC-electrophoresis of peat mud diluted by sulfate sodium-calcium mineral water Uvinskaya of moderate mineralization. The dynamics of clinico-endoscopic and morphological picture of the disease was controlled with fibrogastroduodenoscopy with target biopsy of the periulcer zone and antral stomach, investigation of secretory and acid-producing function of the stomach, hormone profile and collagen metabolism. Positive changes were found attributed to favourable shifts in the hormone status and normalization of collagen metabolism in duodenal mucosa.

Dynamics of formation of lysoforms on enzymatic hydrolysis of phosphatidylalkanols.

Hydrolysis of 1,2-dioleoyl-sn-glycero-3-phosphomethanol (DOPM), 1,2-dioleoyl-sn-glycero-3-phosphoethanol (DOPEt), 1,2-dioleoyl-sn-glycero-3-phosphoethyleneglycol (DOPEG), and 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) catalyzed by phospholipase A2 (PLA2) from porcine pancreas was studied in single-component and binary model bilayer membranes (liposomes). The presence of anionic phosphatidylalkanols increases the rate of hydrolysis of zwitterionic DOPC in liposomes by the action of PLA2. The rate of formation of lysoforms of anionic ("acidic") lipids at the initial reaction stage in single-component liposomes increased in the following sequence: DOPEG < DOPM < DOPEt (compared with that for the zwitterionic DOPC). In binary liposomes formation of lyso-DOPC increased in the presence of acidic lipids in the following sequence: DOPM < DOPEt < DOPEG. This indicates that the size of polar fragment of the second substrate and the presence of free hydroxy groups in the "head" of DOPEG may possibly activate DOPC hydrolysis by the action of PLA2 in the presence of anionic phospholipids including cardiolipin; the studied phospholipids model fragments of the latter.

[Main sources of the formation of contingents of tuberculosis patients with drug-resistant or sensitive Mycobacterium tuberculosis isolates]. / Osnovnye istochniki formirovaniia kontingentov bol'nykh tuberkulezom, vydeliaiushchikh lekarstvenno-ustoichivye i chuvstvitel'nye mikobakterii tuberkuleza.

To form contingents of pulmonary tuberculosis patients with isolated drug-resistant Mycobacteria tuberculosis (MBT) strains, chronic patients having the disease for more than 3 years (50.3%) head the list, new cases (36.9%) detected in the past 3 years (1994-1996) rank second, next are those with relapsing tuberculosis (12.8%) identified in 1994-1996. The ratio of the above group of patients with isolated drug-sensitive MBT species is 23.3, 68.8, and 7.9%, respectively.

[Spread of drug-resistant strains of mycobacterium tuberculosis among patients with isolated bacteria]. / Rasprostranenie lekarstvenno-ustoichivykh shtammov mikobakterii tuberkuleza sredi bol'nykh tuberkulezom baketiovydelitelei.

The paper provides the data on the spread of drug-resistant strains of Mycobacterium tuberculosis (MBT) among contingents of patients with pulmonary tuberculosis who have been registered at a dispensary and new cases. The spread was 42.5 and 20.0%, respectively. In the past 3 decades, the spread of primary (baseline) drug resistance of MBT has increased from 6.5 to 22.0%.

[Results of 30 year follow-ups of pulmonary tuberculosis patients contingent with isolated drug resistant or sensitive strains]. / Rezul'taty 30-letnikh nabliudehií za kontingentom bol'nykh tuberkulezom legkikh s vydeleniem lekarstvenno-ustoíchivykh i chuvstvitel'nykh mikobakterií tuberkuleza.

The paper provides the results of follow-ups of 269 patients with pulmonary tuberculosis, 170 and 39 isolated drug-resistant or sensitive Mycobacteria tuberculosis (MT), respectively. Drug resistance to MT was found to lower the feasibility of curing the patients, to prolong their cure, and to slow down the rate of decreasing the cohort of tuberculosis patients registered at the dispensary. In the first 5-years of the follow-up, there were lower death rates due to tuberculosis in a group of patients with drug-resistant MT than in those with drug-sensitive one.

[Present-day problems of detection and patient treatment with tuberculosis]. / Sovremennye problemy vyiavleniia i lecheniia bol'nykh tuberkulezom.

The problems of detection and treatment of patients with tuberculosis, which are of paramount importance for today, are presented through the analysis of the tuberculosis epidemic situation in the country and of the sociomedical characteristics of new cases (49% of socially disadapted persons). Of the most significance is the need to promptly identify patients with strains on their referral to the general somatic hospitals for complaints by using 3-multiple sputum bacterioscopy for Mycobacterium tuberculosis by the Ziehl-Neelsen method. The vital problem is also to change chemotherapeutical regimens as more severe progressive types of the disease require more active treatment in the first months after detection especially in the cohort of socially disadapted persons.