Bulletin of Russian State Medical University

In spite of successful measures taken to reduce mortality associated with tuberculosis, this disease is still widely spread. In some Russian regions the number of patients with tuberculosis is no short of the epidemic level. The long-term use of antibiotics, changes in the composition of the human microbiota and a few other factors have contributed to the emergence of drug-resistant and hypervirulent sublineages of Mycobacterium tuberculosis . Insufficient fundamental knowledge of mechanisms underlying the emergence and evolution of M. tuberculosis clones simultaneously resistant to a wide spectrum of antibiotics and exhibiting increased virulence complicates the situation and necessitates a new strategy to combat the disease. The key concepts of this strategy are «superorganism», «microbiota» and «resistome». The emergence of multidrug-resistant (MDR) and extensively resistant (XDR) strains should be addressed in the context of the «superorganism»; among its components are the human body, its microbiota (specifically, the bacteria that affect the immune status), and M. tuberculosis itself. Clinically studied phenotypes and genotypes of MDR/XDR strains are a result of clonal variability that M. tuberculosis develops as part of this «superorganism». Therefore, it is important to focus on the development of vaccines, adjuvants and probiotics with selective immunomodulating and antioxidant properties.

Ключевые слова

Development of new tuberculosis (TB) vaccines and host-oriented therapy requires understanding mechanisms mediating protective antituberculous immunity. Antigen-specific Th1 lymphocytes have long been considered as the main correlate of TB protection. However, recent data do not confirm this concept. This article discusses debatable issues concerning the role for Th1 lymphocytes in antituberculous immunity, as well as their use as correlates of protection in preclinical and clinical studies assessing the effectiveness of new candidate TB vaccines.

Ключевые слова

The spread of multidrug and extensively drug-resistant Mycobacterium tuberculosis urges the development of novel antituberculosis drugs. Previously, we studied the compounds representing the class of substituted imidazo[1,2- b ][1,2,4,5] tetrazines capable of inhibiting serine/threonine protein kinases (STPK) in the original M. smegmatis aphVIII + test-system. To unveil the mechanism of action of drug candidates, it is necessary to search for mutations in the mycobacterial genome that confer resistance to these compounds. The aim of our work was to find and describe such mutations in M. smegmatis strains. We carried out the whole-genome sequencing of 9 mutants resistant to 3 imidazo[1,2- b ][1,2,4,5]tetrazines. Seven of 9 mutant strains were found to have the Y52H mutation in the highly conserved mycobacterial gene MSMEG_1601 encoding a protein with an unknown function. Additionally, three of those 7 strains were shown to have two mutations in the MSMEG_1380 encoding a transcriptional regulator. The remaining 2 mutant strains had mutations in MSMEG_0641 and MSMEG_2087 genes encoding transporter-proteins. No mutations were found in STPK genes, meaning that they might be not the primary targets of the studied compounds. Further investigation of MSMEG_1601 function may be of interest as this protein might be the biological target or a part of a new mechanism underlying resistance to antituberculosis drug candidates.

Ключевые слова

Evolution of Mycobacterium tuberculosis have lead to the development of a number of lineages that have unique phenotypes and genotypes and are associated with certain geographical regions. Thus, compared to the reference strains of M. tuberculosis , Beijing and LAM genotypic lineages, which are the most common in the world, are highly virulent and transmissible. However, the extensive use of antibiotics over the past 50 years has caused the next evolutionary leap, which yielded new, epidemiologically dangerous sublineages: Beijing-B0 in Russia, Beijing-modern-4 in China and KZN in South Africa. This study aimed at investigating the effect the immune dysfunction predictors registered in patients have on the severity of tuberculosis (TB) developing after contracting M. tuberculosis Beijing-B0. We compiled a sample of patients with newly diagnosed TB caused by M. tuberculosis Beijing-B0, searched for the immune-suppressing diseases/conditions in their medical history and developed their immunograms. No connection was found between the state of the immune system and the characteristics of the disease we considered.

Ключевые слова

Introduced into clinical practice in 2011, non-invasive prenatal testing (NIPT) allows detection of chromosomal aneuploidies in the fetus using maternal blood samples. Multiple studies have shown that one of the key factors affecting the result of this test is the fetal DNA fraction. The aim of this work was to develop a method capable of measuring the fetal DNA fraction based on targeted SNP sequencing. We selected polymorphisms with high frequency of heterozygous genotype from the international HapMap database. To estimate the frequency of these polymorphisms in the Russian population, we used 827 DNA donor samples. Fetal DNA fraction was measured in 87 plasma samples of pregnant women. Sequencing was performed on Ion Proton and Ion S5. We determined the frequencies of the studied polymorphisms in the pooled samples and compared the data on 53 SNPs in the pooled and 87 individual samples. The median difference was 3.4%. The correlation between the results obtained by targeted SNP sequencing and Y chromosome read count was 0.7. Thus, the proposed method can be used to estimate the fetal DNA fraction using SNP genotyping regardless of the fetus’s sex.

Ключевые слова

Cystic fibrosis (CF) is one of the most common monogenic disorders of humans. The knowledge of population frequency of a mutant genotype causing a monogenic disease helps to optimize DNA testing and to reduce its costs and time required for the procedure. This article presents the results of a retrospective study of the CFTR gene in 191 children with mixed manifestations of CF. To screen for 24 most common mutations, we used the diagnostic PCR panel; minor mutations were detected by next generation sequencing. The diagnostic panel allowed us to identify 18 typical CFTR mutations, including F508del (allelic frequency of 54.7%), dele 2,3 (21kb) (7.3%), 2143delT (3.4%), 2184insA (3.4%), 1677delTA (2.4%), N1303K (2.1%), 3849+10kbC>T (2.1%), E92K (2.1%), G542X (1.6%), W1282X (1.6%), S1196X (1.3%), R334W (1.0%), 394delTT(0.8%), 3944delGT (0.8%), 3821delT (0.5%), 2789+5G>A (0.5%), 621+1G>T(0.3%), and 2183AA>G (0.3%). Sequencing revealed the presence of 24 potentially pathogenic CFTR variants in the sample. We also discovered 8 minor CFTR variants previously unseen in Russian patients with CF, including 4 new CFTR mutations: p.Glu819Ter, p.Gln378Ter, p.Val1360Phefs, and p.Lys1365Argfs.

Ключевые слова

Natural non-pathogenic and vaccine strains of human enteroviruses are currently considered as promising agents capable of treating various kinds of cancer, including glioblastoma multiforme, the most aggressive brain tumor with so far no effective therapy. Enteroviruses can selectively replicate in cancer cells and cause tumor lysis. However, the ability of enteroviruses to persist in tumor tissue for a long period of time and to replicate in several successive cycles while spreading from cell to cell remains largely unclear. This study aimed to determine the possibility of completely destroying subcutaneous mouse xenografts of human glioblastomas through a single intravenous administration of virus-carrying peripheral blood leukocytes, as well as to find out the duration of persistence of the virus in the body of experimental animals in the context of viral therapy. Neurospheres were formed in vitro by incubating fragments of patients-derived glioblastomas and used to initiate subcutaneous tumors in immunodeficient mice. It was established that human peripheral blood leukocytes infected in vitro can effectively deliver Coxsackie A7 virus to the tumor cells. A single injection of 2 × 104 virus-infected leukocytes led to a gradual regression of tumors, while the virus presence was constantly detectable in the blood of mice, up to the complete regression of the tumors. The study allows to make the conclusion that blood leukocytes can effectively deliver Coxsackie A7 virus to the tumor. In the absence of a full-fledged immune response in mice, the viruses persist in tumors leading to their complete destruction.

Ключевые слова

Pathogenesis of colorectal cancer (CRC) is accompanied by significant changes in the immune system. However, the role of the adenosine-A2AR-mediated immunosuppressive pathway in oncogenesis and more specifically, the expression of ectonucleoside triphosphate diphosphohydrolase-1 (ENTPD1, also known as CD39) remains unclear. The aim of this work was to study the role of СD4 Т cells, most importantly CD39-expressing regulatory T cells (Tregs) in the formation of immune suppression in CRC and in patients with acute pancreatitis (AP). Expression of CD39 by peripheral blood lymphocytes and tumor-infiltrating lymphocytes (TILs) was measured by flow cytometry. The levels of CD39 messenger RNA (mRNA) in the peripheral blood leukocytes were determined by real-time PCR. Our study reveals that patients with CRC accumulate peripheral CD4CD39 cells in the advanced stages of the disease. The proportion of CD39-expressing CD4 Т cells in the total pool of TILs was higher than in the peripheral blood of the same patients. Tregs of both peripheral blood and tumor specimens of CRC patients showed increased CD39 expression. We have found reliable correlations between the levels of CD4CD39 T cells and the parameters of cell-mediated immunity in CRC patients. Also, CD39 mRNA levels gradually increased during CRC progression. In contrast, patients with AP have the same levels of CD39 mRNA and peripheral blood CD4CD39Т cells as the controls. Finally, we conclude that activation of CD4CD39 Т cells has an important role in oncogenesis and needs to be studied further.

Ключевые слова

Nitric oxide has a significant role in the pathogenesis of bronchial asthma and hypertension. Its synthesis is catalyzed by NO synthases. The nucleotide composition of genes coding for these enzymes can affect their activity; therefore, it is important to understand the effect of the NOS3 786C/T polymorphism (rs2070744) on the blood levels of nitric oxide in patients with bronchial asthma and hypertension. Our study recruited 71 individuals. The main group consisted of 24 asthmatic hypertensive patients. Two comparison groups included patients with isolated asthma and isolated hypertension. All patients were genotyped for the NOS3 786C/T polymorphism. We measured total nitric oxide metabolites in their blood using a photocolorimetric technique and the Griess reagent. The levels of nitric oxide in the exhaled air were determined electrochemically using a portable NObreath monitor. The blood levels of nitric oxide metabolites amounted to 69.7 (60.0; 70.4) μmol/l in the CC genotype carriers, 68.9 (57.7; 77.4) μmol/l in the CT genotype carriers and 67.7 (59.7; 79.3) μmol/l in the patients with the TT genotype ( p = 0.843). Individually, the groups demonstrated a clear association between the NOS3 786C/T polymorphism and the blood levels of nitric oxide metabolites. The patients with bronchial asthma and hypertension demonstrated a tendency to increasing nitric oxide levels following the pattern CC < CT < TT ( p = 0.033 and p = 0.024, respectively). Thus, the C allele of the NOS3 786C/T polymorphism is associated with lower blood levels of nitric oxide metabolites in patients with bronchial asthma and hypertension.

Ключевые слова

Patients with chronic obstructive pulmonary disease (COPD) are unable to do physical exercises included into standard pulmonary rehabilitation programs. Neuromuscular electrical stimulation (NMES) is a good alternative for such patients as it does not aggravate shortness of breath. The aim of this work was to assess the effect of short-term NMES of the quadriceps femoris muscle on the physical activity of patients with COPD. Our prospective open randomized study was carried out in 36 patients distributed into two groups. The main group was administered NMES for 10 days. On day 10 clinical and functional parameters, as well as adverse events, were evaluated. On admission to hospital, the groups did not differ in terms of the studied parameters. Following the treatment course, the main group significantly improved their step count and electromyography results (418.5 (86.0; 815.0) vs. 226.7 (48.0; 660.0), p = 0.02, and 463.0 (122; 804) vs. 210.5 (64; 481), p = 0.0001, respectively). The patients scored much less on the Mmrc and Borg scales and the CAT-test: 22.8 (18.0; 34.0) vs. 28.4 (26.0; 34.0), p = 0.00007; 2.7 (2.0; 4.0) vs. 3.1 (3.0; 4.0), p = 0.03; and 6.3 (5.0; 7.0) vs. 7.2 (6.0; 9.0), p = 0.0002, respectively. No adverse events were registered in the main group. Based on the obtained results, we conclude that shortterm NMES of the quadriceps femoris muscle improves physical activity, the quality of life and ability to do physical exercise in patients with COPD providing them with a good alternative to standard rehabilitation programs.

Ключевые слова

Hypothyroidism remains a pressing concern. Laser irradiation is a widely used treatment option for patients with thyroid pathologies. Its efficacy depends on the applied dose. Changes in the form and volume of the structural components of the glands, such as thyrocytes and follicles, are dose-dependent and signal their functional state, which affects production, accumulation and secretion of thyroid hormones. The aim of our study was to explore the effect of infrared medium-power laser with total energy densities of 112 J/cm2 and 450 J/cm2 on the morphology and function of the thyroid and its follicles in health and hypothyroidism. The experiment was conducted in male rats. It was demonstrated that laser radiation affects the morphological state of thyrocytes and follicles of both intact animals and animals with experimentally induced hypothyroidism. Comparison of two laser regimens revealed that 112 J/cm2 energies stimulated tissue regeneration and thyroid activity in general, whereas 450 J/cm2 energies suppressed those processes. Our findings can be used to study hypothyroidism treatment options in the experimental setting.

Ключевые слова

Iatrogenic arteriovenous fistulas make up only 0.22% of all fistulas. This article reports a postoperative arteriovenous fistula in a female patient who initially presented with a vestibular schwannoma and was operated using the retrosigmoid approach. Undesired clinical symptoms developed after the patient had been discharged home, and included pulsatile tinnitus, which intensified when the patent tilted or turned her head. The diagnosis was established based on cerebral angiography findings during the second hospital stay. This case report describes complications of retrosigmoid craniotomy, clinical manifestations of the arteriovenous fistula and successful fistula embolization.

Ключевые слова

Nevus sebaceous of Jadassohn (NSJ) is a benign skin lesion, a hamartoma typically localized to the face or scalp and equally common in men and women. Pluripotent epithelial cells that give rise to NSJ provide a favorable environment for benign and malignant tumors to form in the nevus. Because of the possibility of malignant transformation, NSJ should be removed after puberty. If surgery is impossible, long-term observation is indicated. In this work we present two cases of successful NSJ treatment with the CO laser in young patients.

Ключевые слова

Ключевые слова