Synthesis, characterization and complex evaluation of antibacterial activity and cytotoxicity of new arylmethylidene ketones and pyrimidines with camphane skeletons

The synthesis of 20 arylidenecamphors and 15 pyrimidines with camphane skeleton is described in the current report. A modified method for preparation of sterically hindered 2- aminopyrimidines in two steps was demonstrated. The evaluation of their in vitro activity against Mycobacterium tuberculosis H37Rv showed different MIC values (up to 0.91 μM for ketone 39). Compound 35 demonstrated moderate (8.23 μM), but sustainable activity toward a collection of drug-resistant M. tuberculosis strains. Many of the compounds (especially among 2-aminopyridines 42–56) exhibited good to excellent activity against different strains of pathogenic bacteria and fungi (MIC up to 0.60 μM for compound 50), compared with reference antibiotics. Many of the newly designed compounds possess also in vitro cytotoxicity.This study was supported by: Bulgarian National Science Fund- project KP-06-H39/7 and Spanish Ministry of Science, Innovation and Universities- Grant RTI2018-094629-BI00. MEDINA’s authors disclosed the receipt of financial support from Fundación MEDINA, a public-private partnership of Merck Sharp and Dohme de EspañaS.A./Universidad de Granada/Junta de Andalucía

ИНДУКЦИЯ АПОПТОЗА КЛЕТОК МЕЛАНОМЫ В16 ПРИ ВОЗДЕЙСТВИИ ПРЕПАРАТА ФАКТОРА НЕКРОЗА ОПУХОЛЕЙ АЛЬФА В СОСТАВЕ ВИРУСОПОДОБНЫХ ЧАСТИЦ IN VITRO

Aim: to evaluate the antitumor activity of the drug containing TNF-alpha and high-polymer double-stranded RNA (dsRNA) in the composition of virus-like particles (VLP-TNF-alpha) on B16-F10 melanoma cells. Material and Methods. Analysis of the anti-proliferative effect of VLP-TNF-alpha as well as its components, TNFalpha and dsRNA, was carried out using the MTT -test. Apoptosis of melanoma cells was assessed by flow cytofluorimetry with FITC-annexin V. Results. It was shown that the cytotoxic effect of the drug containing the combination of TNF-alpha and dsRNA on melanoma cells significantly exceeded the total cytotoxic effect of TNF-alpha or dsRNA alone (LD50 for combination drug was 0.05 μg/ml, TNF-alpha – 9.5 μg/ml, dsRNA>20 μg/ml). Conclusion. The drug containing TNF-alpha and dsRNA molecules may be a promising drug for the treatment of malignant tumors, including melanoma.Цель исследования – оценка противоопухолевой активности препарата, содержащего фактор некроза опухолей альфа (ФНО-альфа) и двуспиральную рибонуклеиновую кислоту (дсРНК) в составе молекулярной конструкции («вирусоподобной частицы») (ВПЧ-ФНО-альфа), на клетках меланомы B16-F10. Материал и методы. Анализ антипролиферативного действия ВПЧ-ФНО-альфа и его компонентов ФНО-альфа и дсРНК проводили с помощью МТТ-теста, апоптоз клеток меланомы оценивали методом проточной цитофлуориметрии с ФИТЦ-аннексином V. Результаты. Показано, что токсическое воздействие препарата, содержащего ФНО-альфа и дсРНК в составе вирусоподобных частиц, на клетки меланомы значительно превышает суммарный токсический эффект ФНО-альфа и дсРНК в отдельности (ЛД50 составляет, соответственно, для комплексного препарата 0,05 мкг/мл, ФНО-альфа – 9,5 мкг/мл, дсРНК >20 мкг/мл). Заключение. Препарат, содержащий ФНО-альфа и дсРНК в молекулярной конструкции, может быть многообещающим терапевтическим средством для лечения злокачественных новообразований, включая меланому

PREVALENCE OF HIGH CARCINOGENIC RISK HUMAN PAPILLOMAVIRUS IN THE REPUBLIC OF SAKHA (YAKUTIA), ST. PETERSBURG AND THE REPUBLIC OF KARELIA

Results of the study in three regions of the Russian Federation have shown widespread of high carcinogenic human papillomavirus among patients of dermatovenerological and gynecological profile. Detection of viral DNA in the material from the cervix and urethra ranged from 25,2 (Karelia) to 42,5 (Sakha Republic (Yakutia) per 100 examined patients. In all areas in 2010-2011 first place was occupied by 16-th virus genotype - from 11,5 (Sakha Republic (Yakutia) to 15,9 (St. Petersburg) per 100 patients. Prevalence of 11 other types differs. In the Sakha Republic (Yakutia) the second rank place was occupied by types 31 and. 51 (8,0 per 100 examined patients), in St. Petersburg - by 56 and 31 types (9,7 and. 7,6 per 100 patients). Age risk group contains patients of the age of 20-29 years. Information on circulating genotypes of the virus is a necessary part of surveillance to validate vaccination against human papillomavirus infection and evaluation of its efficiency

Tuberculosis outcomes related to the Mycobacterium tuberculosis genotype

Mycobacterium tuberculosis strains of different phylogenetic lineages and genetic families differ in biological properties that determine, to some extent, epidemiological features and clinical manifestation in tuberculosis (TB) patients.The aim of the study was to assess the risk of an adverse outcome of the disease in TB patients caused by various M. tuberculosis genotypes.Materials and methods. A total of 425 patients with respiratory TB were enrolled in this study. They were registered at phthisiatric facilities in the Omsk region from March 2015 to June 2017 period and included: males — 73.1%, mean age 39.9 years, females — 26.9%, mean age 42.0 years. M. tuberculosis culture and drug susceptibility testing and DNA extraction were performed in accordance with standard methods. Strains were assigned to the M. tuberculosis Beijing genotype and its epidemiologically relevant clusters B0/W148 and 94-32 by PCR based detection of specific markers. Non-Beijing strains were subjected to spoligotyping.Results. We found that 66.5% isolates belonged to the Beijing genotype, 12.8% — to LAM, 10.1% — to T, and 4.7% — to the Ural genotype. Multi-drug resistance (MDR) to anti-TB drugs was observed in 195 M. tuberculosis strains (45.9%). Moreover, Beijing genotype was more often isolated from patients with MDR-TB infection (PR = 2.09 (95% CI 1.6–2.74) and TB infection associated with HIV infection (PR = 1.14 (95% CI 1.01–1.31). Lethal outcome was double higher in patients infected with Beijing vs. non-Beijing strains, 28.6% vs. 14.0% (PR = 2.03; 95% CI 1.3–3.17). The risk factors were identified as follows: young age 18–44 years (RR = 1.7; 95% CI 1.18–2.7), co-morbidity with HIV (RR = 5.0; 95% CI 3.39–7.45), multiple (RR = 1.7; 95% CI 1.14–2.55) and extensive drug resistance (RR = 2.57; 95% CI 1.35–4.92), and association with the Beijing genotype (RR = 2.0, 95% CI 1.3–3.17).Conclusion. M. tuberculosis spread in the Omsk region is characterised by significant prevalence of the Beijing genotype, associated with multiple and extensive drug resistance. A significant association of adverse clinical outcomes and various factors, including association with the Beijing genotype, requires development of new approaches in the fight against tuberculosis

Molecular epidemiological monitoring of the tuberculosis pathogen in the Arkhangelsk region

Introduction. Against the background of improvement of the main epidemiological indicators (morbidity and mortality) for tuberculosis in the Arkhangelsk region, the proportion of newly diagnosed tuberculosis patients with multidrug-resistant pathogen (MDR-TB) increased from 18.7% in 2002 to 33.8% in 2018. The purpose of this study was the genotypic characterization of Mycobacterium tuberculosis strains obtained from newly diagnosed tuberculosis patients in the Arkhangelsk region in 2018. Materials and methods. 89 M. tuberculosis strains isolated in 2018 from newly diagnosed tuberculosis patients were studied. Beijing genotype, its clusters B0/W148 and Central Asian/Russian were determined by PCR detection of the specific markers: IS6110 insertions in the dnaA-dnaN region, mutations in codons 48 of the mutT4 gene (CGG GGG) and 58 of the mutT2 gene (GGA CGA), IS6110 insertions in the Rv2664 region-Rv2665 and Rv1359-Rv1360, substitutions G A in the sigE gene. Non-Beijing strains were spoligotyped. Results. Drug resistance was detected in 41.6% (37/89), MDR — in 33.7% of strains. In 90% (27/30) of MDR strains, resistance to rifampicin and isoniazid was due to rpoB Ser531Leu and katG Ser315Thr mutations. Following M. tuberculosis genotypes were identified: Beijing (67.4%), T (14.6%), Ural (4.5%), Haarlem (4.5%), LAM (2.3%) and CAS1-Delhi (1.1%). Among the Beijing strains, clusters Central-Asian/Russian (60%; 36/60) and B0/W148 (30%; 18/60) prevailed. The majority of MDR strains belonged to the Beijing family (93.3%; 28/30), of which 64.3% (18/28) and 21.4% (6/28) belonged to clusters B0/W148 and Central-Asian/Russian, respectively. Conclusion. In heterogeneous population of the causative agent of tuberculosis in the Arkhangelsk region, the most common strains were those of the Beijing genotype; in 2018 its share increased to 67.4% (40.4% in 1998–1999). Among MDR strains, the proportion of Beijing reached 93.3%, of which more than half (64.3%) belonged to the epidemiologically and clinically significant in Russia cluster B0/W148

Genome-Wide Mycobacterium tuberculosis Variation (GMTV) Database: A New Tool for Integrating Sequence Variations and Epidemiology

Background Tuberculosis (TB) poses a worldwide threat due to advancing multidrug-resistant strains and deadly co-infections with Human immunodeficiency virus. Today large amounts of Mycobacterium tuberculosis whole genome sequencing data are being assessed broadly and yet there exists no comprehensive online resource that connects M. tuberculosis genome variants with geographic origin, with drug resistance or with clinical outcome. Description Here we describe a broadly inclusive unifying Genome-wide Mycobacterium tuberculosis Variation (GMTV) database, (http://mtb.dobzhanskycenter.org) that catalogues genome variations of M. tuberculosis strains collected across Russia. GMTV contains a broad spectrum of data derived from different sources and related to M. tuberculosis molecular biology, epidemiology, TB clinical outcome, year and place of isolation, drug resistance profiles and displays the variants across the genome using a dedicated genome browser. GMTV database, which includes 1084 genomes and over 69,000 SNP or Indel variants, can be queried about M. tuberculosis genome variation and putative associations with drug resistance, geographical origin, and clinical stages and outcomes. Conclusions Implementation of GMTV tracks the pattern of changes of M. tuberculosis strains in different geographical areas, facilitates disease gene discoveries associated with drug resistance or different clinical sequelae, and automates comparative genomic analyses among M. tuberculosis strains

Genotyping of Mycobacterium tuberculosis isolate, nonBeijing group, circulating in Ural region

A total of 80 Mycobacterium tuberculosis isolates, nonBeijing genotype from newly detectedTB patients, living in Ural region of Russia, were studied by using MIRU-VNTR typing and spoligotyping.Mutations associated with resistance to isoniazid and rifampicin were detected by microchip technology («TB-Biochip»). The discrimination powers of 15 locus MIRU-VNTR genotyping and spoligotyping were 0,99 and 0,97 respectively. Some MIRU-VNTR clusters may contain isolates with different spoligotypes and may differ in drug mutation. Predominant groups of isolates accordingto MIRU-VNTR data were LAM(31,25%) nUral(26,25%).Методами MIRU-VNTRn сполиготипирования проведено генотипирование 80 изолятов Mycobacterium tuberculosis гpyппы nonBeijing, выделенных от вновь выявленных больных туберкулезом легких - жителей Уральского региона. Мутации, обуславливающие лекарственную устойчивость к рифампицину и изониазиду, были определены тест-системой «ТБ-Биочип». Дискриминирующие способности 15-локусного MIRU-VNTR типирования и сполиготипирования составили 0,99 и 0,97 соответственно. Некоторые VNTR-кластеры могут содержать изоляты различающиеся сполиготипами и спектром мутаций устойчивости к рифампицину и изониазиду. Среди изолятов non-Beijing, по данным MIRU-VNTR типирования,преобладали представители генетических групп: LAM(31.25%) и Ural(26,25%)

Случай хромосомно-интегрированного вируса герпеса человека 6В типа у часто длительно болеющего ребенка

The five years old boy with recurrent respiratory tract infections was under observation of infectiologist due to high levels of human herpes virus type 6 (HHV-6), found in patient’s blood and saliva during a few years. The patient got the medicines against the HHV-6, without any effect. We investigated the patient’s and his mom’s nails and found the HHV-6 type B, so it was inherited chromosomal integrated HHV-6. Thus, to avoid the unnecessary treatment, in case of repeated high level of HHV-6, we need to exclude chromosomal integrated HHV-6.У часто и длительно болеющего пятилетнего мальчика при обследовании выявлена высокая концентрация вируса герпеса человека 6 (ВГЧ-6) в слюне и крови (более 1×106 копий ДНК в мл). По этому поводу он наблюдался инфекционистом и получал противовирусную терапию в течение нескольких лет без эффекта. В ногтевых пластинах пациента и его матери методом ПЦР обнаружена ДНК ВГЧ-6В в количестве 6,37 lg и 6,04 lg на 105 клеток соответственно, что свидетельствует о хромосомно-интегрированной форме ВГЧ-6. При высокой концентрации в крови или повторном выявлении ВГЧ-6 необходимо исключать хромосомно-интегрированную форму инфекции во избежание необоснованного лечения

Genotypic and phenotypic characteristics of <i>Mycobacterium tuberculosis</i> drug resistance in TB children

Background. Russian Federation is included in the list of 30 countries with the highest burden of tuberculosis, including MDR tuberculosis. The most important part of this problem is the primary MDR/XDR TB in children.The aim: a comparative analysis of the phenotypic and genotypic profile of drug resistance to anti-tuberculosis drugs (ATP) according to whole genome sequencing of M. tuberculosis strains from children.Materials and methods. Whole genome sequencing (WGS) results of 61 M. tuberculosis isolates from children with tuberculosis in 2006–2020 in the Russian Federation were analyzed for anti-TB drug resistance mutations, according to the WHO catalog and were compared with the results of phenotypic drug sensitivity.Results. The M. tuberculosis belonged to two genetic groups: Beijing genotype – 82 % (50/61) dominant Central Asian Russian (31/50) and B0/W148 (16/50) subtypes, and non-Beijing (Ural, S, LAM) – 18 % (11/61). Three isolates belonged to Asian Ancestral subtype (3/50). Of the 61 isolates, only 14.7 % (9/61) were sensitive to antiTB drugs, 49.2 % (30/61) were MDR and 14.7 % (9/61) were pre-XDR. Comparison of the resistance profile (MDR/pre-XDR) with genotype revealed an upward shift for Beijing isolates, in particular Beijing B0/W148 (15/16) subline compared to other Beijing (19/34) (Chi-square with Yates correction = 5.535; p &lt; 0.05) and nonBeijing (5/12) (Chi-square with Yates correction = 6.741; p &lt; 0.05) subtypes. Discrepancies between genotypic and phenotypic drug resistance profiles were found in 11.5 % (7/61) of cases.Conclusions. Based on the analysis of WGS data, the genotypic characteristics of M. tuberculosis and the most complete set of drug resistance mutations were obtained, indicating a significant prevalence in MDR and pre-XDR TB of cases caused by epidemic subtypes of Beijing (B0/W148 and Central Asian Russian). The molecular mechanisms of adaptation of M. tuberculosis to the treatment of anti-TB drugs are not unique for the child population but reflect the general processes of the spread of MDR/XDR in Russia