Protein Tyrosine Phosphatase CD45 As an Immunity Regulator and a Potential Effector of CAR-T therapy.

The leukocyte common antigen CD45 is a receptor tyrosine phosphatase and one of the most prevalent antigens found on the surface of blood cells. CD45 plays a crucial role in the initial stages of signal transmission from receptors of various immune cell types. Immunodeficiency, autoimmune disorders, and oncological diseases are frequently caused by gene expression disorders and imbalances in CD45 isoforms. Despite extensive research into the structure and functions of CD45, the molecular mechanisms behind its role in transmitting signals from T-cell receptors and chimeric antigen receptors remain not fully understood. It is of utmost importance to comprehend the structural features of CD45 and its function in regulating immune system cell activation to study oncological diseases and the impact of CD45 on lymphocytes and T cells modified by chimeric antigen receptors.

[Vocal fold polyps: modern possibilities of laser surgery in comparison with the use of cold microinstruments]. / Polipy golosovykh skladok: sovremennye vozmozhnosti lazernoi khirurgii v sravnenii s ispol'zovaniem kholodnykh mikroinstrumentov.

OBJECTIVE: To perform a comparative analysis of phonosurgical interventions, which were made using a 445 nm semiconductor laser or cold microinstrumentation in patients with «vocal fold polyp¼ diagnosis. MATERIAL AND METHODS: The number of surgical interventions equal 30 regarding vocal fold polyp was done in otorhinolaryngology clinic of Pavlov First State Medical University of St. Petersburg in the period from September 2021 yr. to September 2022 yr. The phonosurgical intervention was performed under the conditions of direct suspension microlaryngoscopy by Kleinsasser under general anesthesia using high frequency ventilation through an endotracheal catheter. Surgical interventions were carried out using a 445-nm semiconductor laser in pulsed mode for patients of the 1st group (n=15) and using microsurgical instruments for patients of the 2nd group (n=15). The comparative analysis was conducted based on the results of an objective vocal function assessment by acoustic analysis, as well as on the data of a subjective assessment of the voice using the questionnaire VHI-10rus. The severity of reactive inflammatory phenomena in the postoperative period has been compared and the oscillating movements of vocal folds have been evaluated according to the videolaryngoscopy data. The severity of pain syndrome in the postoperative period was assessed by visual analogue scale. RESULTS: There were no statistically significant differences in the results between two groups. Similarly, there were no statistically significant differences in parameter recovery degree, evaluated by videolaryngoscopy data. Moreover, a full recovery of mucosal wave was found in 93% of patients operated with laser on 7th day, while 47% of patients had the same result after use of cold microinstrumentation. CONCLUSION: The use of 445-nm semiconductor laser for phonosurgical interventions in vocal folds' polyps can be recommended for predicted successful recovery of vocal function, minimization of reactive inflammatory phenomena in postoperative period and early patients' rehabilitation.

BAL fluid analysis in the identification of infectious agents in patients with hematological malignancies and pulmonary infiltrates.

The present study aims to evaluate the diagnostic yield of bronchoalveolar lavage (BAL) fluid in patients with hematological malignancies and describe the most common pathogens detected in BAL fluid (BALF.) An analysis of 480 BALF samples was performed in patients with hematological malignancies over a period of 7 years. The results of culture methods, PCR, and immunoenzymatic sandwich microplate assays for Aspergillus galactomannan (GM) in BALF were analyzed. Further, the diagnostic thresholds for Aspergillus GM and Pneumocystis jiroveci were also calculated. Microbiological findings were present in 87% of BALF samples. Possible infectious pathogens were detected in 55% of cases; 32% were classified as colonizing. No significant difference in diagnostic yield or pathogen spectrum was found between non-neutropenic and neutropenic patients. There was one significant difference in BALF findings among intensive care units (ICU) versus non-ICU patients for Aspergillus spp. (22% versus 9%, p = 0.03). The most common pathogens were Aspergillus spp. (n = 86, 33% of BAL with causative pathogens) and Streptococcus pneumoniae (n = 46, 18%); polymicrobial etiology was documented in 20% of cases. A quantitative PCR value of > 1860 cp/mL for Pneumocystis jirovecii was set as a diagnostic threshold for pneumocystis pneumonia. The absorbance index of GM in BALF of 0.5 was set as a diagnostic threshold for aspergillosis. The examination of BAL fluid revealed the presence of pathogen in more than 50% of cases and is, therefore, highly useful in this regard when concerning pulmonary infiltrates.

Reduction of Thymoglobuline from 7.5 mg/kg to 6 mg/kg in conditioning regimen extended time to the first cytomegalovirus detection after allogenic haematopoietic stem cell transplantation.

INTRODUCTION: The optimal dosage of anti-thymocyte globulin (ATG) may influence the outcome of patients after allogenic haematopoietic stem cell transplantation (HSCT). The aim of our study was to analyse human cytomegalovirus (CMV) infection data, incidence of graft-versus-host disease and other clinical endpoints comparing two patients cohorts that were administered two different Thymoglobuline Genzyme doses as part of the HSCT conditioning regimen. MATERIALS AND METHODS: Total of 65 adult patients received ATG (7.5 mg/kg or 6 mg/kg) as a part of the fludarabine/busulfan/ATG conditioning regimen. CMV DNAemia was monitored after HSCT using quantitative real-time PCR and preemptive treatment was started for viral loads above 1000 cp/ml. RESULTS: The mild ATG dose reduction extended the time to the first CMV detection after transplantation (28 days for 7.5 mg/kg dose vs. 40 days for 6 mg/kg dose, p = 0.04). But it did not reduce the incidence or influence first anti-CMV treatment onset, the initial viral load, peak viral load in whole blood or the antiviral therapy parameters (all p 0.18). No impact of ATG dose reduction on incidence of graft-versus-host-disease, relapse of underlying disease or mortality within first year after transplantation (all p 0.32) were observed. CONCLUSIONS: The reduced ATG dosages can allow lower toxicity of conditioning regimen while keeping the performance.

[Fibrinolytics: from the thrombolysis to the processes of blood vessels growth and remodeling, neurogenesis, carcinogenesis and fibrosis].

One of the most outstanding scientific achievements in the thrombolysis is the development and administration of fibrinolysin - the first Soviet drug that lyses blood clots. Intracoronary administration of fibrinolysin reduced the mortality of patients with myocardial infarction by almost 20%. For his work in this field Yevgeny Chazov was awarded the Lenin Prize in 1982. Over the next decades, under his leadership, the Cardiology Center established scientific and clinical laboratories that created new generations of drugs based on fibrinolytics for treating patients with myocardial infarction, restoration of blood flow in ischemic tissue, and also studying the mechanisms of remodeling of blood vessels involving the fibrinolysis system. It have been found new mechanisms of regulation of the navigation of blood vessels and nerves growth, tumor growth and its metastasis with the participation of the fibrinolysis system proteins. The review reports the role of the fibrinolysis system in the thrombolysis, blood vessels growth and remodeling, neurogenesis, carcinogenesis and fibrosis. The article is dedicated to the 90th anniversary of academician E.I. Chazov.

Multifaced Roles of the Urokinase System in the Regulation of Stem Cell Niches.

Proliferation, subsequent migration to the damaged area, differentiation into appropriate cell types, and/or secretion of biologically active molecules and extracellular vesicles are important processes that underlie the involvement of stem/progenitor cells in the repair and regeneration of tissues and organs. All these functions are regulated through the interaction between stem cells and the microenvironment in the tissue cell niches that control these processes through direct cell-cell interactions, production of the extracellular matrix, release of extracellular vesicles, and secretion of growth factors, cytokines, chemokines, and proteases. One of the most important proteolytic systems involved in the regulation of cell migration and proliferation is the urokinase system represented by the urokinase plasminogen activator (uPA, urokinase), its receptor (uPAR), and inhibitors. This review addresses the issues of urokinase system involvement in the regulation of stem cell niches in various tissues and analyzes the possible effects of this system on the signaling pathways responsible for the proliferation, programmed cell death, phenotype modulation, and migration properties of stem cells.

[Variability and Identification Power 60 X-Cromosome in Two Native Siberian Populations].

Genetic diversity of 60 X-chromosome single nucleotide polymorphisms (XSNPid panel) in populations of Siberian Tatars and Tuvinians is described. A close spectrum of allele frequencies and a low level of their genetic differentiation (Gst = 0.021) is revealed. High discriminating power of the XSNPid panel in populations under study is demonstrated. The random matching probability (MP) of multilocus genotypes in males is 1.12 x 10⁻¹8 in Siberian Tatars and 7.77 x 10⁻¹6 in Tuvans. In females, MP is several orders of magnitude lower: 1.51 x 10⁻²5 in Siberian Tatars and 1.83 x 10⁻²³ in Tuvinians.

[PARTICIPATION OF UROKINASE RECEPTOR AND ITS ENDOGENOUS LIGANDS IN BRAIN DEVELOPMENT AND FORMATION OF COGNITIVE FUNCTIONS].

Recently it has been found that the urokinase receptor (uPAR) and its ligands - urokinase (uPA) and SRPX2 protein play an important role in the development and functioning of the brain. There is a strong association between uPAR gene polymorphism and autism disorders in humans. Patients with autism, intractable lobe epilepsy, verbal dyspraxia and perisylvian polymicrogyria display significant changes in uPAR expression. Mice, lacking the uPAR gene develop epilepsy and demonstrate abnormal social behavior. uPA and SRPX2 protein, have been shown to be involved in pathological brain conditions such as autism, cognitive deficits and language disorders. Urokinase system that stimulates blood vessel growth as demonstrated before, also plays an important role in the regulation of the nerve growth via matrix remodeling and activation of neurotrophic and angiogenic factors. Moreover, the urokinase system also functions as a guidance system which determines the growth trajectory of the vessels' and nerves' in tissue regeneration. This review summarizes and integrates the results and recent progress in the field of uPAR and its endogenous ligands in brain development and cognitive functions.

[Ganciclovir treatment failure in adult allogeneic hematopoietic stem cell transplant recipients with cytomegalovirus infection--a single centre experience]. / Klinická rezistence lidského cytomegaloviru pri lécbe gancyklovirem u pacientu po alogenní transplantaci hematopoetických bunek--zkusenosti jednoho centra.

OBJECTIVE: To determine the incidence of infection with ganciclovir-resistant cytomegalovirus (CMV) in adult allogeneic hematopoietic stem cell transplant (HSCT) recipients. Clinical resistance or treatment failure was defined as persistent DNAemia or increasing viral load in peripheral blood after 2 weeks of virostatic treatment. The association between the treatment failure and viral resistance was analysed. The presence of ganciclovir-resistant CMV strains was confirmed by genotypic testing able to detect mutations conferring resistance. METHODS: In 2012 and 2014, 40 patients who underwent allogeneic HSCT for hematologic malignancies and were treated for human CMV reactivation/disease were followed up prospectively. In patients with treatment failure, CMV DNA was isolated and analysed by nucleotide sequence analysis of the UL 97 and UL 54 genes conferring resistance to the virostatic agent. RESULTS: The treatment failure occurred in seven patients, but ganciclovir resistance conferring mutations were only detected in two of them (mutations L595F and M460I in the UL 97 gene). Another mutation in the UL 97 gene (N510S) was found in a patient with recurrent CMV replication who needed to be retreated but did not meet the criteria for treatment failure. CONCLUSION: The low incidence of genetically confirmed ganciclovir-resistant CMV isolates in HSCT recipients with relatively common clinical treatment failure suggests that the mechanism underlying slower viral clearance is often other than mutations conferring ganciclovir resistance to the virus.

[Opposite trends in the incidence of gonorrhea and syphilis in the East Bohemian Region - 30 years of surveillance]. / Protikladný vývoj trendu incidence kapavky a syfilis ve východoceském regionu - 30 let surveillance.

The epidemiology of selected sexually transmitted diseases in the Czech Republic has been carefully evaluated for many years. Data from 1981-2011 for eastern Bohemia shows a sharp decrease in the incidence of gonorrhea in 1993-1994 and a very low incidence thereafter with a slightly higher prevalence in males. However, syphilis and genitourinary infections with Chlamydia trachomatis show entirely opposite trends. Also, for the similar number of diagnostic tests performed, Chlamydia had a 10 fold higher rate of positive results. This underscores the changing epidemiology of sexually transmitted infections (STI) and necessity for adapting the reporting algorithms accordingly.

Viral gastroenteritis in Eastern Bohemia Region of the Czech Republic.

PURPOSE: Acute gastroenteritis is one of the most common diseases in humans worldwide and represents a significant cause of mortality and morbidity. The majority of cases are of viral aetiology, evidence for which has been increasing in the past decade. Several studies on the prevalence in European countries of viral aetiology of gastroenteritis have been published in the last decade, but none from the Czech Republic. MATERIAL AND METHODS: In total 107 faeces samples obtained from patients hospitalised in the University Hospital in Hradec Králové were examined by immunochromatographic tests using ROTA-ADENO Card Rapid-Viditest (VIDIA, Czech Republic) and RidaQuick Norovirus (R-Biopharm, Germany), and by an in-house Real-time PCR panel. RESULTS: Overall findings of viruses detected by PCR in the tested faeces samples were: rotaviruses in 29.9%, noroviruses in 14.0% and adenoviruses in 5.0%. Immunochromatographic antigen detection performed at lower sensitivity compared with PCR: rotaviruses in 28.0%, noroviruses in 4.7% and adenoviruses in 2.0%. Our findings demonstrate even lower sensitivity of the used immunochromatographic tests compared with manufacturers data. CONCLUSION: Our study has revealed limitations in immunochromatographic tests, especially in their sensitivity and the necessity for using another confirmatory method. We have set up real-time PCR in routine diagnosis of viral gastroenteritis in our hospital.

Influenza in seasons 2009-2013 in the Faculty Hospital Hradec Kralove, East Bohemia.

PURPOSE: The evaluation of four post-pandemic influenza seasons 2009-2013 in the Faculty hospital Hradec Králové and comparison of used rapid antigen tests (RATs) with real time RT-PCR. MATERIAL AND METHODS: Between November 2009 and June 2013 were examined 3845 samples from patients with respiratory tract infections by RATs (Influenza A/B 2 Panel Test (GECKO® Pharma, Germany), Rapid VIDITEST Influenza A+B Card (VIDIA®, Czech Republic) and BinaxNOW Influenza A&B Test (ALERE®, USA) or real time RT-PCR (RTR InfA/H1N1 Detection Set (Roche®), RealStar® Influenza S and T RT-PCR Kit 3.0 (Altona ®, Germany). RESULTS: A totally 1059 samples were examined simultaneously by RAT and real time RT-PCR. The overall sensitivity and specificity of RATs compared with real time RT-PCR were 32,2 % and 98,1 % for influenza A and 17,6 % and 99,4 % for influenza B. Higher sensitivity of RATs were in children (66,6 %) compared with adults (14,3 - 40,0 %). In the first three post-pandemic seasons were continuously decrease of positive samples from 23,5 % in season 2009-2010 to 3,3% in season 2011-2012, but in season 2012-2013 were rapidly in-crease of positive results, to 31,5%, with high share of influenza -A/H1N1/2009 (79,6%). CONCLUSION: Our results shown insufficient sensitivity of all used RATs and necessity of having other confirmatory test, like RT-PCR. It was also shown unexplained increase of case and influenza severity in season 2012-2013.

[RNA editing. The classical cases and the outlook of new technologies].

This article provides an overview of studied RNA editing cases and the examples of RNA editing investigations that apply to different experimental data. Typical RNA editing site prediction errors and the methods to minimize them are shown. The outlook of up-to-date technologies and further RNA editing studies are discussed.

[Genetic diversity and relationships of northern eurasia populations for polymorphic Alu-insertions].

We for the first time have examined the autosomal gene pool of the Siberia, Central Asian and the Far East populations (27 populations of 12 ethnic groups) using a set of polymorphic Alu insertions in the human genome. The results of the analysis testify (i) to a significant level of genetic diversity in the Northern Eurasian populations and (ii) to a considerable differentiation of gene pool in the population of this region. It has been shown that at the CD4 locus, the frequency of Alu (-) is inversely related to the Mongoloid component of the population, the lowest and highest frequencies of the Alu deletion at locus CD4 were recorded respectively in Eskimo (0.012) and Russian and Ukrainian (0.35). The analysis of gene flow proved Caucasoid populations (Russian, Tajik and Uzbek), as well as those of Turkic ethnic groups from the Southern Siberia (Altaians and Tuvinians) and Khanty and Mansy populations to be the recipients of a considerable gene flow from the outside of the concerned population system, as compared with the East Siberian and the Far East ethnic groups. The results of the correlation analysis received with use polymorphic Alu insertion testify to the greatest correlation of genetic distances with anthropological characteristics of populations.

[Cytomegalovirus and its relationship to chronic inflammatory bowel diseases and tumors]. / Cytomegalovirus a jeho vztah k chronickým zanetum strev a nádorovým onemocnením.

Human cytomegalovirus (hCMV), a member of the Betaherpesvirinae subfamily, was long considered an important pathogen especially in immunocompromised hosts, mainly in post-transplant patients receiving immunosuppressive therapy. Results of many studies from the last decade suggest that cytomegalovirus is a very important pathogen having a greater association with many other diseases. The relationship of hCMV to chronic inflammatory bowel diseases, ulcerative colitis and Crohn's disease, has been long studied but results of the studies have been rather inconclusive. Interestingly, hCMV was found to be more prevalent in lesions of severe ulcerative colitis, in particular ulcerative colitis refractory to immunosuppressive therapy (40-57% in results from clinical studies) than in mild or moderate colitis (up to 5% in most studies). Another group of diseases possibly related to hCMV is tumors. Numerous studies discuss the oncomodulatory role of hCMV in malignant glioblastoma, colorectal adenocarcinomas, prostate cancer and lymphomas. Despite extensive evidence suggesting a possible relationship of hCMV to some tumors, such as successful induction of malignant carcinoma by hCMV latent infection in animal models, hCMV has not been classified as an oncovirus. However, its potential in this respect is apparent.

[The use of IgG antibody avidity assays in the diagnosis of cytomegalovirus infection]. / Vyuzití testu avidity IgG protilátek v diagnostice cytomegalovirové infekce.

Human cytomegalovirus (CMV) is the most common cause of congenital infection. Primary CMV infection can lead to severe disease and complications in patients immunocompromised as a result of disease or therapy. IgG antibody avidity assays make it possible to differentiate between primary infection and reactivation of latent infection or reinfection. The study objective was to determine CMV IgG avidity by enzyme-linked immunosorbent assay (ELISA) with denaturation of IgG antibody binding to the antigen and by chemiluminiscent microparticle immunoassay (CMIA) on an Abbott Architect analyzer. Both methods yielded comparable CMV IgG avidity results. In some cases, the Abbott test was superior in reflecting IgG antibody maturation during primary infection to microplate ELISA using antigen-antibody complex dissociation by a denaturing agent.

[Tick-borne encephalitis in the East Bohemia Region and its microbiological diagnostic pitfalls]. / Klíst'ová encefalitida ve vychodoceském regionu a úskalí její mikrobiologické diagnostiky.

AIM OF THE STUDY: The analysis of cases of tick-borne encephalitis (TBE) laboratory diagnosed at the Department of Virology of the Institute of Clinical Microbiology of the University Hospital in Hradec Králové in 2003-2006 and comparison of the data from 2003-2006, 1999-2002 and 1995-1998. Another objective was to compare two routinely used ELISA (enzyme-linked immunosorbent assay) diagnostic kits for TBE. MATERIAL AND METHODS: The samples from 3284 patients tested by the Department of Virology of the University Hospital in Hradec Králové in 2003-2006 were analyzed. The detection of the specific IgM and IgG antibodies in serum and cerebrospinal fluid specimens was performed using two ELISA diagnostic kits (Euroimmun, Germany, and Test-Line Ltd, Clinical Diagnostics, Czech Republic). Non-specific reactivity of IgM antibodies was only analyzed in 2004-2006. RESULTS: In 2003-2006, TBE was diagnosed in 163 patients of the University Hospital in Hradec Králové aged from 3.5 months to 80 years. The number was by 48 cases (29%) higher than that in 1995-1998 and by 36 cases (22%) higher than that in 1999-2002. TBE was confirmed in 94 males (57.7%) and 69 females (42.3%). More than half of the patients were aged above 41 years (94/163, i.e., 57.6 %) and 60 patients (37 %) were older than 51 years. A biphasic course of infection was observed in 93/136 cases (68.4%). As many as 101/136 patients (74 %) had a history of a tick bite, one patient reported the consumption of non-pasteurized milk. Thirty-eight (1.4 %) specimens showed non-specific reactivity of IgM antibodies. None of the cases showed cerebrospinal fluid reactivity in the absence of serum reactivity. CONCLUSIONS: Increase in the diagnosed TBE cases was reported in 2003-2006 in the East Bohemia Region. It was due mainly to more severe clinical forms of TBE that were more frequent than the milder ones. The distribution of the confirmed TBE cases remained unchanged in terms of age and sex, but the seasonal peak moved from summer to autumn. ELISA proved suitable for use in the routine diagnosis of TBE. Both of the tested commercial kits are of good quality. Nevertheless, in view of possible non-specific reactivity of IgM antibodies, IgG antibodies need to be tested in both the first acute serum specimen and paired sera. The detection of antibodies in cerebrospinal fluid is unlikely to be of diagnostic benefit for TBE. Close cooperation between the attending physician and clinical microbiologist is crucial.

[A new yeast strain for brewery: properties and advantages].

Beer is a natural product and is a multicomponent system that has both positive and negative consumer properties. Organoleptical off-flavors of beer are difficult to eliminate. Yeasts are the main active component of the system. The relationship between beer quality and yeast usage is well known. New industrial strains for brewery are continuously developed. An industrial yeast Saccharomyces cerevisiae strain was obtained and showed high technological properties, including efficient fermentation, a reduced production of sulfur hydrate, and a high diacetyl reduction rate. The advantages made it possible to develop new brands of beer and nonalcoholic products. The commercial use of the strain was patented. The strain was deposited in the Russian Collection of Industrial Microorganisms.

[Regulation of growth and remodeling of blood vessels: the unique role of urokinase].

Urokinase-plasminogen activator (uPA) is a multifunctional fibrinolytic protein activating growth factors, inducing proteolytic cascades, modulating cytokines, regulating receptor shedding, cellular phenotypic modulation and protein expression. These mechanisms underlie the ability of uPA to stimulate the key processes of vascular remodelling, atherosclerosis progression, restenosis and angiogenesis, -- cell migration and proliferation. We summarized data received by us and others concerning the role of uPA in blood vessel remodelling and growth. At the present stage, the uPA may be considered as a perspective target for influences directed on both the prevention of negative arterial remodelling and restenosis as well as the stimulation of vessel growth at ischemic diseases.