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.

[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.

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.

[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.

Prevention of infection transmission during stem cell transplantation.

Group of 152 patients (investigated before autologous transplantation) and 35 healthy donors for allogeneic transplantation was examined for the risk of infection transmission that can be associated with the infusion of cryopreserved peripheral blood progenitor cells to the patient and/or cross-contamination of stored grafts. No laboratory signs of active infection were found in 22 donors (63 %) and in 91 patients (60%). The most common was active infection by herpes viruses--50 cases in patients, 21 cases in donors; hepatitis B was found in only two cases. The rate of clinically unsuspected (but dangerous) infections in donors and patients thus remains relatively high in spite of the fact that the system of donor search and the whole transplantation procedure have improved in the last years. The system of safety assurance is extremely important and the whole palette of preventive tests according to EBMT (European Blood and Marrow Transplantation Group) and ISHAGE (International Society for Hemotherapy and Graft Engineering) is fully justified.

Potential risk of infection transmission during storage and transplantation of hematological progenitor cells. Safety assurance.

In a group of 71 patients and 22 donors the danger of infection transmission by infusion of cryopreserved peripheral blood progenitor cells to the patient and/or cross contamination of stored grafts was evaluated. No laboratory signs of active infection were found in 15 donors (13 related, 2 unrelated; 68%) and in 55 patients (77%). Active infection by herpesviruses was the most common (in 13 patients and 7 donors), hepatitis B being found in only one case. The cytomegalovirus IgG test was the most common marker of previous infection; it was found in 14 donors and 55 patients. The rate of clinically unsuspected infections in donors and patients including cases requiring immediate treatment among the patients group is relatively high and fully justifies the practice of prophylactic serological testing in the whole range of tests according to the European Blood and Marrow Transplantation Group and International Society for Hematotherapy and Graft Engineering in both autologous and allogeneic transplantations of hematopoietic stem cells.

[Involvement of urokinase and its receptor in the remodeling of normal and pathological tissue]. / Urokinaza i ee retseptor kak uchastniki strukturnoi perestroiki tkanei v norme i pri patologii.

Urokinase is the main component of the fibrinolytic system and comprises two functions. Namely, the catalytic domain enables to trigger the proteolytic cascade in blood and in the vessel wall, stimulating the activation of growth factors, matrix remodelling and smooth muscle cell proliferation. On the other hand, the amino terminal domain of urokinase that is homologous to epidermal growth factor promotes urokinase to bind to the specific receptor. The cell surface binding of uPA may initiate the activation of the second messengers, protein phosphorylation and cell migration. This dual function of urokinase affects vascular growth, as well as the intimal thickening of the vessel wall after intravascular injury. Also uPA and its receptor are implicated in many physiological and pathological processes, including ovulation, embryogenesis, cancer metastasis, inflammation, and wound healing.

Urokinase plasminogen activator induces smooth muscle cell migration: key role of growth factor-like domain.

We defined the role of urokinase plasminogen activator (uPA) and its growth factor-like domain (GFD) in stimulating smooth muscle cell (SMC) migration. Recombinant uPA (r-uPA) stimulated migration approximately 3-fold whilst the recombinant uPA mutant containing an altered GFD (r-uPAmut) was ineffective. Both uPA variants bound to the same high affinity receptor in a competitive manner. FGF-2- and PDGF-BB-induced migration was also dependent on uPA, their effects being antagonized by addition of a uPA-neutralizing antibody or the r-uPAmut. Thus r-uPA is chemotactic for SMC and stimulation of cell migration by PDGF-BB and FGF-2 is dependent on uPA. The GFD of uPA is essential for its chemotactic effects.

[Effect of physical load deficiency on the system of calcium transport in the sarcoplasmic reticulum of skeletal muscles amd myocardium]. / Vliianie fizicheskikh nagruzok na sistemu transporta kaltsiia v sarkoplazmaticheskom retikulume skeletnykh myshts i miokarda.

The authors have obtained data characterizing the changes in Ca-transporting function of sarcoplasmatic reticulum (SR) of the skeletal muscles varying in type of muscle fibers as well as the cardiac muscle of the rats during their suspension at an angle of inclination of the upper end of the body of 30 degrees and with the gravity load relief of the hind limbs. Making wide use of the available literature data on the role of Ca2+ as a secondary mediator of cellular homeostasis the authors discuss the possible mechanisms of changing the function of Ca-transporting system of the reticulum directed, by this means, the way of further studies of the transport function of Ca-ATP-ase of the membranes of SR while investigating the effects of weightlessness.

[Ca2+ transport by the skeletal muscle sarcoplasmic reticulum in rats after suspension of the hindlimbs]. / Transport Ca2+ sarkoplazmaticheskim retikulumom skeletnykh myshts krys pri vesovoi razgruzke zadnikh konechnostei.

The functional capacity of skeletal muscle SR was examined in the lateral gastrocnemius (LG) of rats submitted to 1-40-day of disuse produced by a hindlimb suspension (HS). The Ca uptake by SR was measured in homogenate of muscle by means fo Ca-selective electrode, and the time course of SR Ca(2+)-pump inactivation in tissue homogenates incubated at 45 degrees C was studied. There were po significant effects of hindlimb unweighting on muscular Ca pump during 1-14 days of HS. The amount of Ca uptake in SR of LG was about 41% higher in the HS group than in the control after 25 days of HS. After 40 days of HS, the activity of Ca(2+) transport increased by 58% in SR atrophied HS muscle compared with the control. The increased resistance of LG muscle SR Ca pump to thermal exposure was observed after HS. The possible reasons of muscular Ca-pump activation after HS were discussed.

[Clinical and functional results of aortocoronary shunting in patients with ischemic heart disease]. / Kliniko-funktsional'nye rezul'taty aortokoronarnogo shuntirovaniia u bol'nykh ishemicheskoi bolezn'iu serdtsa.

Survival rates were evaluated in 3 groups of patients: those with an isolated lesion of the anterior interventricular branch of the left coronary artery (group 1), multiple coronary arterial lesions (group 2) and a more than 50% narrowing of the left coronary-arterial trunk (group 3) following autovenous aortocoronary shunting of 1 to 5 coronary arteries. Drug-treated patients with similar coronary arterial lesions and clinical manifestations were taken as controls. Long-term survival rates of surgical patients were significantly higher, as compared to those of medication-treated patients in groups 2 and 3. There was no significant difference in survival rates of first-group patients. Long-term survival was dependent on the scope of revascularization and pretreatment myocardial contractility.