Anus perinei ventralis in adulthood - case report.

Anorectal malformations present a type of the most serious congenital malformations, either in terms of treatment or treatment outcomes. Anorectal atresia can be subdivided into three categories: the supralevator form, the intermediate type of atresia and the low translevator type. One of the clinical forms of low translevator type in girls is a perineal fistula opening just behind the vaginal entrance on the perineum, with a fully developed sphincter complex dorsally from the fistula (so called anus perinei ventralis). The golden standard of surgical treatment of anus perinei ventralis in children is Peñas procedure, which was used as a guideline for anorectal reconstruction in our adult patient, as well.

Antimetastatic Effect of Liposomal Recombinant Lactaptin.

Antimetastatic effect of the liposomal form of recombinant lactaptin RL2 (a proteolytic fragment of human breast milk κ-casein; 8.6 kDa) was studied in A/Sn mice after intravenous transplantation of GA-1 tumor with high rate of liver metastases. Tumor growth in the liver was found in all mice. In animals dying early, the tumors were presented by multiple nodes of about the same size; in mice dying later, the tumors in the liver were presented by just few large nodes formed by cells that survived chemotherapy. A single intravenous injection of RL2 lactaptin in liposomes prolonged lifespan of animals with liver metastases of GA-1 tumor by 1.5 times in comparison with that in untreated animals.

Salivary, gingival crevicular fluid and serum levels of ghrelin and chemerin in patients with periodontitis and overweight.

BACKGROUND AND OBJECTIVE: Nutrition and body weight are modifying factors for periodontitis. The purpose of this study was to quantify two molecules (ghrelin and chemerin), released in association with food intake and obesity, in periodontally healthy and diseased individuals with respect to different body mass categories. MATERIAL AND METHODS: The two main groups (patients with chronic periodontitis and periodontally healthy/gingivitis volunteers) were subdivided into groups of subjects with normal weight [body mass index (BMI) <25] and groups of overweight/obese subjects (BMI ≥25). Subgingival bacteria were analysed and the levels of acylated and total ghrelin, chemerin and interleukin-1ß (IL-1ß) were assessed in saliva, gingival crevicular fluid and serum. RESULTS: The amount of Treponema denticola present subgingivally was significantly higher in the groups of patients with chronic periodontitis as well as in periodontally healthy/gingivitis individuals with BMI ≥25 than in periodontally healthy/gingivitis individuals with BMI <25. The amount of total ghrelin in gingival crevicular fluid differed significantly between the groups, with the lowest levels found in the group of patients with chronic periodontitis and BMI ≥25. The levels of chemerin in gingival crevicular fluid were significantly higher in each chronic periodontitis group than in periodontally healthy/gingivitis individuals with BMI <25. However, the level of IL-1ß in the gingival crevicular fluid was most differentiating between the groups, with the highest levels found in the group of patients with chronic periodontitis and BMI <25 and the lowest levels in periodontally healthy/gingivitis individuals with BMI <25. No significant differences between any groups were seen for chemerin or for acylated ghrelin in the stimulated whole saliva, or for acylated and total ghrelin in peripheral blood serum. The BMI correlated with the serum level of chemerin. CONCLUSION: Low ghrelin and high chemerin levels in the gingival crevicular fluid might be linked to periodontal disease and overweight/obesity. However, unlike IL-1ß, the levels of chemerin and ghrelin in gingival crevicular fluid are not reliable indicators of periodontal destruction.

Modern Approaches for Identification of Modified Nucleotides in RNA.

This review considers approaches for detection of modified monomers in the RNA structure of living organisms. Recently, some data on dynamic alterations in the pool of modifications of the key RNA species that depend on external factors affecting the cells and physiological conditions of the whole organism have been accumulated. The recent studies have presented experimental data on relationship between the mechanisms of formation of modified/minor nucleotides of RNA in mammalian cells and the development of various pathologies. The development of novel methods for detection of chemical modifications of RNA nucleotides in the cells of living organisms and accumulation of knowledge on the contribution of modified monomers to metabolism and functioning of individual RNA species establish the basis for creation of novel diagnostic and therapeutic approaches. This review includes a short description of routine methods for determination of modified nucleotides in RNA and considers in detail modern approaches that enable not only detection but also quantitative assessment of the modification level of various nucleotides in individual RNA species.

[Immunogenicity of recombinant analog of antitumor protein lactaptin].

Therapeutic monoclonal antibodies and recombinant proteins including cytokines are commonly used in the treatment of cancer and inflammatory diseases. In most cases, these protein-based drugs exhibit a high therapeutic efficacy, which is unfortunately frequently associated with a variety of side effects. We have investigated the in vitro and in vivo immunogenicity of recombinant antitumor protein lactaptin (RL2). Based on the qRT-PCR analysis, we have shown that, in MDA-MB-231 human breast adenocarcinoma cells, RL2 suppresses the NF-kB signaling cascade that regulates the reactions of innate immunity. RL2 inhibits the expression of the CXCL1 protein and apoptosis inhibitor A20 and enhances expression of IkB, NF-kB repressor. The ELISA method has been used to evaluate the antibody titer in the blood of mice, which received single and triple intravenous or intraperitoneal injections of RL2. The multiplex immunoassay of 23 cytokines in the mice blood has shown that the RL2 injections lead to a slight increase in the levels of systemic pro-inflammatory cytokine interleukin-5 (IL-5) and keratinocyte chemoattractant (KC), a homologue of human macrophage inflammatory protein-1 (MIP-1). These observations indicate the low immunogenicity of the recombinant lactaptin analog, which can be considered to be a potential molecular drug candidate for further clinical development.

[The risks of retroperitoneoscopic adrenalectomy]. / Rizika retroperitoneoskopické adrenalektomie.

INTRODUCTION: Minimally invasive adrenalectomy has become the gold standard for surgery of the suprarenal gland. Retroperitoneoscopic adrenalectomy with dorsal approach is preferred. The aim of our case report is to discuss potential complications that may arise from retroperitoneoscopic adrenalectomy, specifically an intra-operative injury of the inferior vena cava. CASE REPORT: A 47-year-old male patient was admitted to undergo elective adrenalectomy on the right side. The reason for the surgery was a hormonally active adenoma with clinical signs of Conns syndrome. Biochemistry revealed the typical signs of hyperaldosteronism. A one-year history of unsuccessful treatment for hypertension was known. Ultrasound examination showed an enlarged suprarenal gland on the right side with the diameter of 5.2 cm. A CT scan confirmed the results of the ultrasound examination. Retroperitoneoscopic adrenalectomy was performed. The inferior vena cava was intraoperatively injured. The high pressure in the retroperitoneal space prevented bleeding. The injury to the vena cava was treated using a continuous stitch without the necessity of conversion to open surgery. The patient was discharged on the third postoperative day without any other complications. CONCLUSIONS: Retroperitoneoscopic approach is regarded by many authors as the new gold standard for adrenalectomy. However, very serious complications such as an injury of the inferior vena cava may occur. It is possible to treat this injury using retroperitoneoscopy. The risk of air embolization due to elevated pressure in the retroperitoneum (20 mm Hg) and open lumen of the IVC needs to be taken seriously.Key words: adrenalectomy - retroperitoneoscopy - complication.

[Complete minimally invasive Ivor-Lewis esophageal resection]. / Kompletní miniinvazivní Ivor-Lewisova resekce jícnu.

INTRODUCTION: Minimally invasive esophagectomy is becoming a standard procedure in the treatment of esophageal cancer. We would like to present our experience with Ivor Lewis esophagectomy completed by minimally invasive technique. METHODS: The primary aim of the study was to analyse potential technical difficulties and intraoperative complications of thoracolaparoscopic Ivor Lewis esophagectomy with intrathoracic anastomosis. A secondary aim of the study was to evaluate postoperative complications according to the Clavien-Dindo classification. The inclusion criterion for the study was a history of thoracolaparoscopic esophagectomy. Multidisciplinary approach was employed in the diagnosis, surgery indications and perioperative care of all patients. RESULTS: Between January 2011 and January 2016, 19 patients underwent completely minimally invasive esophagectomy with intrathoracic anastomosis. There were 13 men and 6 women. Adenocarcinoma was confirmed by histopathological examination in all the patients. The cumulative postoperative morbidity was 68.4%. According to the Clavien-Dindo classification, we recorded grade I complications in 10.5% of the patients, grade II in 15.8%, grade III in 36.8% and grade IV in 5.3% of the patients. Anastomotic leak was the most serious complication; it was initially managed by negative pressure (vacuum) therapy followed by stent implantation. Postoperative mortality was 0%. Mean hospital stay was 12 days and mean stay at intensive care unit was 3.6 days. CONCLUSIONS: The basic oncosurgical principles of radicality need to be respected during minimally invasive procedures. However, functionality, safety, and cost effectiveness have to be preserved as well. In this paper, we present thoracolaparoscopic Ivor Lewis esophagectomy as one of feasible and accessible options of intrathoracic anastomosis. It seems to be safe with respect to technical obstacles, short-term and long-term complications.Key words: esophagectomy - intrathoracic - anastomosis - laparoscopy - thoracoscopy.

The role of maternal nutrition, metabolic function and the placenta in developmental programming of renal dysfunction.

The intrauterine environment is critical for the development of the foetus. Barker and colleagues were the first to identify that adverse perturbations during foetal development are associated with an increased risk of developing diseases in adulthood, including cardiorenal disease. Specifically for the kidney, perturbations in utero can lead to nephron deficits and renal dysfunction by a number of mechanisms. Altered programming of nephron number is associated with an increased risk of developing kidney disease via glomerular hypertrophy and reduced vasodilative capacity of the renal blood vessels; both of which would contribute to hypertension in adulthood, with males being more susceptible to disease outcomes. Additionally, alterations in the renin-angiotensin system (RAS) such as an upregulation or downregulation of specific receptors, depending on the nature of the insult, have also been implicated in the development of renal dysfunction. Sex-specific differences in the expression of the RAS during late gestation and in the early postnatal environment have also been identified. Extensive research has demonstrated that both uteroplacental insufficiency and maternal malnutrition alter renal development in utero. Equally, exposure to maternal diabetes and maternal obesity during development are also associated with an increased risk of developing renal disease, however, the mechanism behind this association is poorly understood. Therefore, identifying the link between an adverse intrauterine environment and the programmed kidney disease risk in adulthood may facilitate the development of strategies to alleviate the epidemics of cardiorenal disease worldwide, in addition to understanding why males are more susceptible to adult-onset cardiovascular diseases.

[Primary malignant small bowel tumors]. / Primární zhoubné nádory tenkého streva.

INTRODUCTION: Small bowel presents 75% of the gut length and 90% of the gut surface. However, primary malignant tumors of the small bowel represent only 1-3% of all malignant gastrointestinal tumors. The aim of the present paper is to offer a current review of primary malignant small bowel tumors - their epidemiology, localization, symptoms, diagnostic and treatment options. METHODS: The authors have performed a comprehensive review of databases Medline, Scopus and Google Scholar focusing on studies regarding small bowel cancer. RESULTS: The most frequent small bowel tumors are adenocarcinoma (30-40%), neuroendocrine tumors (35-44%), lymphomas (10-20%) and gastrointestinal stromal tumors (12-18%). Symptomatology is non-specific and varies widely, which is why small bowel cancer is usually diagnosed in a locally advanced stage of the disease. Diagnosis is determined through standard methods (gastroscopy, colonoscopy, CT) and complementary special diagnostic modalities (capsule enteroscopy, enteroscopy, octreotide scan, etc.). Diagnostic process with a negative outcome frequently leads to diagnostic laparoscopy/laparotomy.The treatment of small bowel cancer in patients operated in acute settings is done according to acute abdomen management guidelines. Elective surgery of small bowel cancer differs with respect to the tumor type. Adenocarcinomas and neuroendocrine tumors should be treated with surgical R0 resection with radical lymphadenectomy (and multivisceral resection if necessary). Patients with GIST should undergo en bloc resection with 2-3cm safety resection margins (lymphadenectomy is not necessary). Palliative resection of neuroendocrine tumors can be associated with a significant clinical effect. On the other hand, palliative resection of adenocarcinomas of GIST is not advocated. CONCLUSION: Small bowel cancer is an infrequent condition. Symptoms are non-specific; patients are often diagnosed in an advanced stage of the disease. Achieving R0 surgical resection is usually difficult due to locally advanced stage of the disease. Besides the tumor type, patients prognosis is influenced by very late diagnosis of the tumor. KEY WORDS: primary tumor - small intestine - diagnostics - treatment options - surgical resection.

Light scattering microscopy with angular resolution and its possible application to apoptosis.

An inverted microscope has been modified for light scattering experiments with high angular resolution in combination with transmission, wide-field fluorescence or laser scanning microscopy. Supported by simulations of Mie scattering, this method permits detection of morphological changes of 3T3 fibroblasts on apoptosis and formation of spherically shaped cells of about 20 µm diameter, in agreement with visual observation. Smaller sub-structures (e.g. cell nuclei) as well as cell clusters may possibly contribute to the scattering behaviour. Results of 2-dimensional cell cultures are confirmed by 3-dimensional multicellular spheroids of 3T3 fibroblasts and HeLa 2E8 cervix carcinoma cells, where in most cases no morphological changes are discernable. This offers some advantage of light scattering microscopy for label-free detection of apoptosis and may represent a first step towards label-free in vivo diagnostics.

[The influence of recombinant nucleophosmin 1 on artificial RNA internalization into human adenocarcinoma MCF-7 cells].

In this study we obtained and characterized the recombinant analogue of multifunctional nucleolar phosphoprotein nucleophosmin 1 (NPM1) involved in crucial cellular processes such as transcription, reparation and mitosis. The influence ofnucleophosmin 1 on extrcellular RNAs accumulation in human adenocarcinoma cells MCF-7 was analyzed. It was found that incubation of AluY RNA (n > 300 nt), U24 snoRNA analogues (n ~ 80 nt) with Npm1-His6 resulted in RNA-protein non-covalent complexes formation, but not in case of the short oligoribonucleotide (n = 22 nt). It was shown that interaction of AluY RNA analogue with Npm1-His6 significantly increases transfection efficacy of the RNA into MCF-7 human cells. Altogether, these data allow us to conclude, that nucleophosmin 1 not only binds RNA with complex secondary structure, but also promotes uptake and internalization of such RNA by human cells.

[Impact of anastomotic leakage on oncological outcomes after rectal cancer resection]. / Vliv dehiscence anastomózy na onkologické výsledky u resekcních výkonu pro karcinom rekta.

INTRODUCTION: The aim of the study was to determine the impact of anastomotic leakage on long-term outcomes after curative surgery for rectal cancer. MATERIAL AND METHODS: The study included 174 patients who underwent elective, potentially curative open or laparoscopic resection with anastomoses for rectal cancer at the Department of Surgery of the University Hospital Ostrava from 1 January 2001 to 31 December 2009. Anastomotic leakage was defined as clinically or radiologically confirmed signs of local or diffuse peritonitis, gas, pus or stool from the drain, rectoscopy signs of anastomotic insufficiency, or rectovesical or rectovaginal fistula. The Cox proportional hazards model with forward selection was used to determine the influence of predefined baseline characteristics on overall, disease-free survival and recurrence. The results are presented as Hazard Ratio (HR) with 95% Confidence Interval (CI). RESULTS: Based on the Cox model, anastomotic leakage was not identified as a factor with a significant impact on overall or disease-free survival. Anastomotic leakage, however, has remained an independent risk factor for a higher local recurrence rate (Hazard Ratio: 6.621, 95% CI 1.289-34.020, p=0.024). On the contrary, anastomotic leakage was not identified as a statistically significant prognostic factor for the incidence of distant metastases. CONCLUSION: Anastomotic leakage represents an independent risk factor for a higher local recurrence rate after curative resection for rectal cancer.

EGFR Suppression Inhibits the Sphere Formation of MCF7 Cells Overexpressing EGFR.

The epidermal growth factor receptor (EGFR) is an oncogenic tyrosine kinase that is involved in tumor initiation and progression, making EGFR inhibitors and monoclonal antibodies to this receptor essential for anti-tumor therapy. We have previously shown that EGFR transgene expression in the human breast adenocarcinoma cell line MCF7 (MCF7-EGFR) stimulates the 3D spheroid-like growth. The primary focus of our present work was to investigate whether EGFR inhibition could affect the assembly of spheroids or lead to the destruction of pre-existing spheroids. We compared the effects of anti-EGFR siRNA, the anti-EGFR monoclonal antibody cetuximab, and the tyrosine kinase inhibitor AG1478 on dissociated and spheroid MCF7-EGFR cells. MCF7-EGFR cells were found to have a 2.5-fold higher sensitivity towards the cytotoxic effects of cetuximab and AG1478 compared with the parental MCF7 cell line. The suppression of EGFR mRNA with siRNA was found to reduce the sphere formation, whereas treating the pre-existing spheroids had no such effect. Treatment of dissociated spheroids with cetuximab and AG1478 was also found to inhibit the MCF7-EGFR sphere formation. We suggest that EGFR expression is important, at least, during the spheroid formation stage. The transition of a MCF7wt adherent cell culture to MCF7-EGFR spheroids was accompanied by a considerable increase in N-cadherin adhesion proteins. The level of N-cadherin decreased when MCF7-EGFR cells were treated with siRNA and cetuximab. Thus, we have demonstrated that N-cadherin is involved in the EGFR-dependent formation of MCF7-EGFR spheroids. Accordingly, MCF7-EGFR spheroids can be considered a suitable model for studying aggressive hormone-positive breast tumors.

Light exposure and cell viability in fluorescence microscopy.

Test systems for measuring cell viability in optical microscopy (based on colony formation ability or lysosomal integrity) were established and applied to native cells as well as to cells incubated with fluorescence markers or transfected with genes encoding for fluorescent proteins. Human glioblastoma and Chinese hamster ovary cells were irradiated by various light doses, and maximum doses where at least 90% of the cells survived were determined. These tolerable light doses were in the range between 25 J cm⁻² and about 300 J cm⁻² for native cells (corresponding to about 250-3000 s of solar irradiance and depending on the wavelength as well as on the mode of illumination, e.g. epi- or total internal reflection illumination) and decreased to values between 50 J cm⁻² and less than 1 J cm⁻² upon application of fluorescent markers, fluorescent proteins or photosensitizers. In high-resolution wide field or laser scanning microscopy of single cells, typically 10-20 individual cell layers needed for reconstruction of a 3D image could be recorded with tolerable dose values. Tolerable light doses were also maintained in fluorescence microscopy of larger 3D samples, e.g. cell spheroids exposed to structured illumination, but may be exceeded in super-resolution microscopy based on single molecule detection.

[Tuberculosis outbreak in 13 people in Saxony-Anhalt: indications of an infection chain by spoligotyping]. / Tuberkulose-Ausbruch bei 13 Personen in Sachsen-Anhalt: Hinweis auf die Infektkette durch Spoligotyping.

BACKGROUND: In Germany the number of new tuberculosis incidents continues to decline slightly. Cases of illnesses are often diagnosed late, due to the uncharacteristic course of tuberculosis. The success in environment investigations by health authorities in finding incidents within one infection chain depends on the compliance of patients among other factors. Tuberculosis occurs especially in groups with social disadvantages. In this population segment the identification of contact persons of a patient with infectious tuberculosis presents difficulties. Moreover, identifying the source of infection is frequently not successful. For these reasons, additional infections and sicknesses are caused. The cultural pathogen proof, including type differentiation and resistance testing, continues to be the "gold standard" in tuberculosis diagnostics and is required for the examination of outbreaks. Molecular methods for fine typification of isolated bacterial strains and their subsequent comparison constitute an important tool in the infection epidemiology investigation of incident clustering. OBJECTIVE AND METHOD: The objective of this work is the description of a tuberculosis infection chain in Saxony-Anhalt, which was discovered by means of a molecular biological method. The conservative method represents the environment investigations for the expedient selection of contact persons. The investigations always take 2 directions in the process. On the one hand, it is searched for the still unknown source of infection. On the other hand, persons are to be identified, who have been infected before the diagnosis became known. The diagnostics for contact persons includes besides the lung X-ray checkup exam, the Mendel Mantoux tuberculin skin test (THT), sputum tests and the Interferon Gamma (INF-γ) tests (IGRA). In described case, the molecular biological method of spoliogotyping was employed as additional instrument. Therein, fingerprint maps of the patients' tuberculosis strings were created and compared. Infection chains resulting from the environment investigations were thereby secured. RESULTS: We report on a tuberculosis outbreak in a district of the state of Saxony-Anhalt, which occurred in the time from June 2007 to May 2010. As result of the environment investigations, an infection chain of 13 ill patients (77% ♂, 23% ♀) was identified. In 11 cases the correlation of the infection chain was proven by spoligotyping (9 ♂, 2 ♀). 2 cases (1 ♂, 1 ♀) without verification of the pathogen cannot be attributed on basis of the epidemiological correlations.

[Nuss miniinvasive procedure for pectus excavatum in adolescents and adults]. / Miniinvazivní operace pectus excavatum u adolescentu a dospelých dle Nusse.

Pectus excavatum is a congenital chest wall deformity with depression of the sternum and adjacent costal cartilages. Severe forms of this deformity lead not only to psychosocial deprivation but also limit physical performance due to lung volume reduction and cardiac compression. Open surgical correction using stemochondroplasty represented the gold standard of surgical treatment of pectus excavatum. Miniinvasive technique of corrective steel bar insertion was published in 1998. Since then, so called Nuss operation has become widely accepted. Good experience with this type of the pectus excavatum correction have encouraged us to adopt this procedure. We use this technique not only in children and adolescencents but also in adults suffering from depressed anterior chest wall. We present our initial experience with the treatment of nine patients. We describe the benefits and pitfalls of the method which are known to us.

[NOSE (Natural Orifice Specimen Extraction) in laparoscopic colorectal surgery]. / NOSE (Natural Orifice Specimen Extraction) v kolorektální chirurgii.

INTRODUCTION: The aim of this study was to asses our initial first experience with NOSE techniques in laparoscopic colorectal surgery with both transanal and transvaginal extraction. MATERIAL AND METHODS: In this prospective study, the authors analyzed data from patients in whom NOSE laparoscopic sigmoid, rectosigmoid and rectal resections were performed in the Department of Surgery, University Hospital Ostrava, from May 2011 to October 2011. A group of 7 patients was analyzed based on demographic characteristics (sex,age and BMI). Tumor localization, type of extraction (transanal/transvaginal), the number of removed lymph nodes, tumor size, histology and length of the specimen were also assessed. Furthermore, the following intraoperative data were evaluated: duration of the procedure, frequency of intraoperative complications and conversion rate. During the postoperative period, duration of hospitalization and morbidity rates were evaluated. RESULTS: The patient group included 2 male (28.6%) and 5 female (71.4%) subjects, their median age was 70 years (61-80), BMI 26,76 (24.76-34.67). The pathology was located in the sigmoid colon in 4 cases (57.1%) and in the proximal rectum in 3 cases (42.9%). Transanal extraction was performed in 5 patients (71.4%) and transvaginal extraction in 2 patients (28.6%). The average number of harvested lymph nodes was 13 (10-15), the average lenght of specimen was 16 cm (13-20) and the average tumor size was 4 cm (2-6). Histologically, adenocarcinoma was confirmed in 6 cases (85.7%), and low grade adenoma in 1 case (14.3%). The median duration of surgery was 205 min (140-300) and no intraoperative complications were recorded. No surgical conversion was required. No postoperative complications occured and the median duration of of hospital stay was 7 days (5-11). CONCLUSION: In the selected group of patients, NOSE technique proved to be a safe technique for laparoscopic colorectal procedures, reducing the risk of incisional complications while maintaining the principles of oncological radicality. Therefore, it may be considered a bridge towards NOTES (Natural Orifice Transluminal Endoscopic Surgery), surgery without scars.

Immunogenic Cell Death in Cancer Therapy.

Apoptosis plays a crucial role in chemotherapy-induced cell death. The conventional theory holding that apoptosis needs to be immunologically silent has recently been revised, and the concept of immunogenic cell death (ICD) has been proposed. This review describes the main features of ICD induction. These ICD markers are important for the effectiveness of anticancer therapy, as well as for basic research into cell death regulation. The mechanism of the "vaccination effect" of dying cancer cells undergoing ICD has been fully described, including the activation of specific antitumor response after re-challenge by the same living tumor cells. This review also discusses the whole set of molecular events attributing cell death to immunogenic type: the exposure of calreticulin and the heat shock protein HSP70 to the outer surface of the cell membrane and the release of the nuclear protein HMGB1 and ATP into the extracellular space. ICD inducers of various nature (chemotherapy drugs, cytotoxic proteins, and oncolytic viruses), as well as physical methods, are classified in the current review.

Cancer-associated fibroblasts and their role in tumor progression.

The stromal elements of a malignant tumor can promote cancer progression and metastasis. The structure of the tumor stroma includes connective tissue elements, blood vessels, nerves, and extracellular matrix (ECM). Some of the cellular elements of the tumor stroma are cancer-associated f ibroblasts (CAFs). The origin and function of CAFs have been actively studied over the past thirty years. CAFs produce collagen, the main scaffold protein of the extracellular matrix. Collagen in the tumor stroma stimulates f ibrosis, enhances the rigidity of tumor tissue, and disrupts the transmission of proliferation and differentiation signaling pathways. CAFs control tumor angiogenesis, cell motility, tumor immunogenic properties, and the development of resistance to chemo- and immunotherapy. As a result of metabolic adaptation of rapidly growing tumor tissue to the nutrients and oxygen deprivation, the main type of energy production in cells changes from oxidative phosphorylation to anaerobic glycolysis. These changes lead to sequential molecular alterations, including the induction of specif ied transcriptional factors that result in the CAFs activation. The molecular phenotype of activated CAFs is similar to f ibroblasts activated during inf lammation. In activated CAFs, alpha-smooth muscle actin (α-SMA) is synthetized de novo and various proteases and f ibronectin are produced. Since CAFs are found in all types of carcinomas, these cells are potential targets for the development of new approaches for anticancer therapy. Some CAFs originate from resident f ibroblasts of the organs invaded by the tumor, while others originate from epithelial tumor cells, which are undergoing an epithelial-mesenchymal transition (EMT). To date, many molecular and metabolic inducers of the EMT have been discovered including the transforming growth factor-beta (TGF-ß), hypoxia, and inf lammation. This review classif ies modern concepts of molecular markers of CAFs, their functional features, and discusses the stages of epithelial- mesenchymal transition, and the potential of CAFs as a target for antitumor therapy.

The Signaling Pathways Controlling the Efficacy of Glioblastoma Therapy.

The resistance of glioblastoma to existing therapies puts limits on quality-of-life improvements and patient survival with a glioblastoma diagnosis. The development of new effective glioblastoma therapies is based on knowledge about the mechanisms governing tumor resistance to therapeutic agents. Virotherapy is one of the most actively developing approaches to the treatment of malignant neoplasms: glioblastoma in particular. Previously, we demonstrated that the recombinant vaccinia virus VV-GMCSF-Lact exhibits in vitro cytotoxic activity and in vivo antitumor efficacy against human glioblastoma. However, the studied glioblastoma cell cultures had different sensitivities to the oncotoxic effect of the virus. In this study, we investigated cancer stem cell (CSC) surface markers in glioblastoma cells with different sensitivities to VV-GMCSFLact using flow cytometry and we assessed the levels of proteins affecting viral entry into cells and virus infection efficiency by western blotting. We showed that cell cultures more sensitive to VV-GMCSF-Lact are characterized by a greater number of cells with CSC markers and a lower level of activated Akt kinase. Akt probably inhibits lactaptin-induced apoptosis in virus-resistant cells. Hence, we suggest that the sensitivity of glioblastoma cells to the oncotoxic effect of VV-GMCSF-Lact is determined by the nature and extent of the disturbances in cell death regulation in various cultures. Further investigation of the factors affecting glioblastoma resistance to virotherapy will test this hypothesis and identify targets for antitumor therapy, combined with VV-GMCSF-Lact.