An HNF4α-miRNA inflammatory feedback circuit regulates hepatocellular oncogenesis.

Hepatocyte nuclear factor 4α (HNF4α) is essential for liver development and hepatocyte function. Here, we show that transient inhibition of HNF4α initiates hepatocellular transformation through a microRNA-inflammatory feedback loop circuit consisting of miR-124, IL6R, STAT3, miR-24, and miR-629. Moreover, we show that, once this circuit is activated, it maintains suppression of HNF4α and sustains oncogenesis. Systemic administration of miR-124, which modulates inflammatory signaling, prevents and suppresses hepatocellular carcinogenesis by inducing tumor-specific apoptosis without toxic side effects. As we also show that this HNF4α circuit is perturbed in human hepatocellular carcinomas, our data raise the possibility that manipulation of this microRNA feedback-inflammatory loop has therapeutic potential for treating liver cancer.

Variation in homeodomain DNA binding revealed by high-resolution analysis of sequence preferences.

Most homeodomains are unique within a genome, yet many are highly conserved across vast evolutionary distances, implying strong selection on their precise DNA-binding specificities. We determined the binding preferences of the majority (168) of mouse homeodomains to all possible 8-base sequences, revealing rich and complex patterns of sequence specificity and showing that there are at least 65 distinct homeodomain DNA-binding activities. We developed a computational system that successfully predicts binding sites for homeodomain proteins as distant from mouse as Drosophila and C. elegans, and we infer full 8-mer binding profiles for the majority of known animal homeodomains. Our results provide an unprecedented level of resolution in the analysis of this simple domain structure and suggest that variation in sequence recognition may be a factor in its functional diversity and evolutionary success.

[performance evaluation of the programs aimed at myocardial infraction prevention by relevant mortality rates].

BACKGROUND: Despite implemented measures and general favourable trend, number of patients with myocardial infarction remains high, younger people are increasingly becoming ill and dying. The study purpose: to estimate age-sex mortality dynamics from acute and recurrent myocardial infarction in adults in Moscow compared to Russia's average in 2007-2021 to reveal patterns of mortality change within the implementation period of state prevention programs. MATERIAL AND METHODS: Standardized adult mortality rates for Moscow and Russia, mean expected age at death within interval 20-85 years and gain in life expectancy when eliminating this cause calculated and analyzed. RESULTS: Moscow morbidity rates for acute and recurrent myocardial infarction are twice lower than Russia's, there are higher reduction rates for both diseases - by 16% and 58% respectively. During study period, mortality from myocardial infarction in men was by 45% higher than in women. In 2007 Moscow male-female difference estimated 3% and in 15 years it became 32% due to faster female mortality reduction. Whereas Russia's average age of death from myocardial infarction in 2021 returned to 2010-2011 levels, then in Moscow during 2020-2021 female rates returned to 2008's and male rates fall out the study period. CONCLUSIONS: Since implementation of the first programs on reducing mortality from chronic non-infectious diseases during the 15 years period morbidity and mortality rates from myocardial infarction reduced in Russia and Moscow in all ages. Acute and recurrent myocardial infarction have rejuvenated both in men and women as a negative effect of the pandemic.

[MORTALITY OF THE ADULT POPULATION OF MOSCOW AND THE RUSSIAN FEDERATION FROM OBESITY-ASSOCIATED CAUSES].

BACKGROUND: Currently, obesity is considered one of the most significant health problems, representing a common chronic disease leading to the development of severe comorbidities, accompanied by the loss of disability-adjusted life years and high mortality. Due to the fact that obesity is one of the leading risk factors for a number of non-communicable diseases, such as diseases of the circulatory system, diseases of the endocrine system and malignant neoplasms. AIMS: assessment of adult mortality dynamics from obesity-associated causes in Moscow compared to the Russian Federation in 2011-2020. MATERIALS AND METHODS: data from the analytic package FAISS (internal use program): standardized mortality rates for population of Moscow and the Russian Federation as a whole. RESULTS: Over the 10-year period under study, mortality in the class of diseases of the circulatory system and malignant neoplasms was decreasing, while mortality from diseases of the endocrine system was increasing. Adult mortality from the diseases of the circulatory system in Moscow reduced by 12%, in the Russian Federation - by 25%. It should be noted, that mortality rate in Moscow (302.5 per 100,000) is significantly (by 34%) lower than in Russia (460.3), at the same time, the rate of mortality reduction over a 10-year period is equal and amounts to 23-25%. Myocardial infarction is the most serious obesity-associated disease characterized by high mortality in the class of the diseases of the circulatory system, it has a declining trend in Moscow and in the Russian Federation as a whole. Adult mortality from the diseases of the endocrine system in Moscow increased by 3 times, and in the Russian Federation - by almost 5 times, while the annual increase during the first year of the pandemic was 88% in Moscow and 24% in Russia. In Moscow, more than a half (66%) of deaths from endocrine causes belongs to non-insulin-dependent diabetes mellitus, in the Russian Federation - about 80%. Compared to the Russia's average, in Moscow mortality rates from neoplasms are lower by 8%, but at the same time, the decline happens at a similar rate (11-12%). CONCLUSIONS: Despite the impact of the COVID-19 pandemic, Moscow demonstrates slowdown of growth of mortality rates from obesity-associated diseases compared to the Russian Federation, which could have been positively affected by prevention programs of noncommunicable diseases and promotion of healthy lifestyles.

[The trends in primary morbidity of adult population of the Russian population during implementation of state programs].

The article presents the results of analysis of indices of primary morbidity of adult population in the Russian Federation during implementation of state programs. The purpose of the study was to study the dynamics of indices of primary morbidity of adult population of Federal districts and subjects of the Russian Federation in 2006-2011 and 2012-2018. The analysis established the subjects of the Russian Federation where the mentioned indices factually remained unchanged or had negative dynamics. Also, the periods of health care reform with positive dynamics of indices of primary morbidity were determined. Besides, reforming process was not always systematic and not all planned tasks were completed that , resulted in significant discrepancies in values of indices of primary morbidity among particular subjects of the Russian Federation. This kind of analysis permits to identify specific directions of improving regional policy in health care.

[The main trends in primary morbidity of population in the subjects of the Russian Federation in 2008-2017].

The analysis of primary morbidity is the most important indicator of detection of population health in every single subject of the Russian Federation in whole Russia. This approach also permits to monitor impact of economic and social processes on the national level. The article presents analysis of primary morbidity in the subjects of the Russian Federation in 2008-2017. The trends are established, the rates of increase and decrease of primary morbidity in every particular subject of the Russian Federation are calculated. The subjects with significant increase of particular classes of diseases according ICD-10 were identified. During last decade, the indices of primary morbidity increased on the whole up to 0.9%. The decrease was registered only in the Central and Privolzhskiy Federal Okrugs (4.3% and -1.6% correspondingly. In the rest of Federal Okrugs increase was registered. The highest level of primary morbidity was registered in the Nenets Autonomous Okrug and the lowest level in the Kabarda-Balkar Republic. Ib 2017, the Altai Kray became a leader in registration of primary morbidity of neoplasms and diseases of endocrine system and the Pensa oblast took the lead in registration of primary morbidity of diseases of cardiovascular system. In 2017, the highest level of morbidity of diseases of respiratory system was registered in the Chukchi Autonomous Okrug and of diseases of ear and mastoid bone in the Republic of North Ossetia - Alania. The increase of morbidity in the mentioned classes of diseases in the particular subjects of the Russian Federation requires consideration of leading specialists and corresponding development of measures concerning morbidity stabilizing.

[The trends of population general morbidity in the subjects of the Central Federal Okrug of the Russian Federation].

The article presents the results of analysis of indices of total morbidity of population of the Central Federal Okrug (CFD) of the Russian Federation in 2010-2017. The significant differences in indices of total morbidity between the CFD subjects in certain ICD-10 classes were established. The indices of total morbidity of population during study period (8 years) in the Central Federal District factually didn't altered, while dynamics of indices in other subjects was characterized by multidirectionality. The gap in levels of total morbidity of population in the subjects was significant: from 115,123.6 per 100,000 of population in the Kursk Oblast to 194,404.1 per 100,000 of population in the Orel Oblast. The significant difference in rates of increase/ decrease of indices is noted. Thus, in Moscow decrease rate made up to 10%, while in the Orel Oblast morbidity increased up to 13.1%. Besides, in 2017, the Orel Region took a leadership in rate of increasing of total morbidity in such classes of diseases as infectious and parasitic diseases (39.3%), diseases of blood and blood-forming organs (49.1%), diseases of endocrine system (59,1%), diseases of nervous system (26.8%), diseases of respiratory system (28.2%), diseases of musculoskeletal system (16%), malformations (56%). It is very likely that this trend developed under influence of demographic situation in the subject due to significant increasing of percentage of people older than able-bodied age. The city of Moscow occupies leading position in decreasing of rate of prevalence of diseases and last but one place in level of total morbidity being inferior only to the Kursk Oblast. Thus, in Moscow was noted the most significant decreasing of morbidity in classes of infectious and parasitic diseases (26%) and diseases of digestive system (20.6%). The diseases of blood and blood-forming organs (235.2 per 100,000 population) and mental disorders (2353.5 per 100,000 population) were registered the less. The main contribution into trends of increasing or decreasing of indices is made by persons aged 18 years and older (74.1%).

CHK1 inhibition in soft-tissue sarcomas: biological and clinical implications.

Background: Inhibition of ChK1 appears as a promising strategy for selectively potentiate the efficacy of chemotherapeutic agents in G1 checkpoint-defective tumor cells such as those that lack functional p53 protein. The p53 pathway is commonly dysregulated in soft-tissue sarcomas (STS) through mutations affecting TP53 or MDM2 amplification. GDC-0575 is a selective ATP-competitive inhibitor of CHK1. Methods: We have performed a systematic screening of a panel of 10 STS cell lines by combining the treatment of GDC-0575 with chemotherapy. Cell proliferation, cell death and cell cycle analysis were evaluated with high throughput assay. In vivo experiments were carried out by using TP53-mutated and TP53 wild-type patient-derived xenograft models of STS. Clinical activity of GDC-0575 combined with chemotherapy in patients with TP53-mutated and TP53 wild-type STS was also assessed. Results: We found that GDC-0575 abrogated DNA damage-induced S and G2-M checkpoints, exacerbated DNA double-strand breaks and induced apoptosis in STS cells. Moreover, we observed a synergistic or additive effect of GDC-0575 together with gemcitabine in vitro and in vivo in TP53-proficient but not TP53-deficient sarcoma models. In a phase I study of GDC-0575 in combination with gemcitabine, two patients with metastatic TP53-mutated STS had an exceptional, long-lasting response despite administration of a very low dose of gemcitabine whereas one patient with wild-type TP53 STS had no clinical benefit. Genetic profiling of samples from a patient displaying secondary resistance after 1 year showed loss of one preexisting loss-of-function mutation in the helical domain of DNA2. Conclusion: We provide the first preclinical and clinical evidence that potentiation of chemotherapy activity with a CHK1 inhibitor is a promising strategy in TP53-deficient STS and deserves further investigation in the phase II setting.

STAT3 activation of miR-21 and miR-181b-1 via PTEN and CYLD are part of the epigenetic switch linking inflammation to cancer.

A transient inflammatory signal can initiate an epigenetic switch from nontransformed to cancer cells via a positive feedback loop involving NF-kappaB, Lin28, let-7, and IL-6. We identify differentially regulated microRNAs important for this switch and putative transcription factor binding sites in their promoters. STAT3, a transcription factor activated by IL-6, directly activates miR-21 and miR-181b-1. Remarkably, transient expression of either microRNA induces the epigenetic switch. MiR-21 and miR-181b-1, respectively, inhibit PTEN and CYLD tumor suppressors, leading to increased NF-kappaB activity required to maintain the transformed state. These STAT3-mediated regulatory circuits are required for the transformed state in diverse cell lines and tumor growth in xenografts, and their transcriptional signatures are observed in colon adenocarcinomas. Thus, STAT3 is not only a downstream target of IL-6 but, with miR-21, miR-181b-1, PTEN, and CYLD, is part of the positive feedback loop that underlies the epigenetic switch that links inflammation to cancer.

Molecular mechanism underlying the regulatory specificity of a Drosophila homeodomain protein that specifies myoblast identity.

A subfamily of Drosophila homeodomain (HD) transcription factors (TFs) controls the identities of individual muscle founder cells (FCs). However, the molecular mechanisms by which these TFs generate unique FC genetic programs remain unknown. To investigate this problem, we first applied genome-wide mRNA expression profiling to identify genes that are activated or repressed by the muscle HD TFs Slouch (Slou) and Muscle segment homeobox (Msh). Next, we used protein-binding microarrays to define the sequences that are bound by Slou, Msh and other HD TFs that have mesodermal expression. These studies revealed that a large class of HDs, including Slou and Msh, predominantly recognize TAAT core sequences but that each HD also binds to unique sites that deviate from this canonical motif. To understand better the regulatory specificity of an individual FC identity HD, we evaluated the functions of atypical binding sites that are preferentially bound by Slou relative to other HDs within muscle enhancers that are either activated or repressed by this TF. These studies showed that Slou regulates the activities of particular myoblast enhancers through Slou-preferred sequences, whereas swapping these sequences for sites that are capable of binding to multiple HD family members does not support the normal regulatory functions of Slou. Moreover, atypical Slou-binding sites are overrepresented in putative enhancers associated with additional Slou-responsive FC genes. Collectively, these studies provide new insights into the roles of individual HD TFs in determining cellular identity, and suggest that the diversity of HD binding preferences can confer regulatory specificity.

Autologous dendritic cells prolong allograft survival through Tmem176b-dependent antigen cross-presentation.

The administration of autologous (recipient-derived) tolerogenic dendritic cells (ATDCs) is under clinical evaluation. However, the molecular mechanisms by which these cells prolong graft survival in a donor-specific manner is unknown. Here, we tested mouse ATDCs for their therapeutic potential in a skin transplantation model. ATDC injection in combination with anti-CD3 treatment induced the accumulation of CD8(+) CD11c(+) T cells and significantly prolonged allograft survival. TMEM176B is an intracellular protein expressed in ATDCs and initially identified in allograft tolerance. We show that Tmem176b(-/-) ATDCs completely failed to trigger both phenomena but recovered their effect when loaded with donor peptides before injection. These results strongly suggested that ATDCs require TMEM176B to cross-present antigens in a tolerogenic fashion. In agreement with this, Tmem176b(-/-) ATDCs specifically failed to cross-present male antigens or ovalbumin to CD8(+) T cells. Finally, we observed that a Tmem176b-dependent cation current controls phagosomal pH, a critical parameter in cross-presentation. Thus, ATDCs require TMEM176B to cross-present donor antigens to induce donor-specific CD8(+) CD11c(+) T cells with regulatory properties and prolong graft survival.

[Luminescence of stable stacking aggregates of adenine and uracil in water].

Luminescence and excitation spectra of the highly luminescent stacking dimers of adenine and uracil in water solutions are studied. By the luminescence excitation spectra method it is shown that the stacking aggregates of adenine and uracil are formed with participation of rare forms of monomer molecules: N7H tautomers of adenine and the uracil molecules in rare forms of hydratation, for example molecules without H-bonds with water. The study of temperature dependence of luminescence intensity of monomers and stacking dimers of uracil has shown that stacking dimers do not dissociate even at 85 degrees C similarly as described earlier for adenine and adenosine. Stable stacking aggregates of nucleic bases are most likely to be the precursors of RNA molecules in chemical evolution. This hypothesis is supported by new data on their stability.

Systematic identification of mammalian regulatory motifs' target genes and functions.

We developed an algorithm, Lever, that systematically maps metazoan DNA regulatory motifs or motif combinations to sets of genes. Lever assesses whether the motifs are enriched in cis-regulatory modules (CRMs), predicted by our PhylCRM algorithm, in the noncoding sequences surrounding the genes. Lever analysis allows unbiased inference of functional annotations to regulatory motifs and candidate CRMs. We used human myogenic differentiation as a model system to statistically assess greater than 25,000 pairings of gene sets and motifs or motif combinations. We assigned functional annotations to candidate regulatory motifs predicted previously and identified gene sets that are likely to be co-regulated via shared regulatory motifs. Lever allows moving beyond the identification of putative regulatory motifs in mammalian genomes, toward understanding their biological roles. This approach is general and can be applied readily to any cell type, gene expression pattern or organism of interest.

Conservation and regulatory associations of a wide affinity range of mouse transcription factor binding sites.

Sequence-specific binding by transcription factors (TFs) interprets regulatory information encoded in the genome. Using recently published universal protein binding microarray (PBM) data on the in vitro DNA binding preferences of these proteins for all possible 8-base-pair sequences, we examined the evolutionary conservation and enrichment within putative regulatory regions of the binding sequences of a diverse library of 104 nonredundant mouse TFs spanning 22 different DNA-binding domain structural classes. We found that not only high affinity binding sites, but also numerous moderate and low affinity binding sites, are under negative selection in the mouse genome. These 8-mers occur preferentially in putative regulatory regions of the mouse genome, including CpG islands and non-exonic ultraconserved elements (UCEs). Of TFs whose PBM "bound" 8-mers are enriched within sets of tissue-specific UCEs, many are expressed in the same tissue(s) as the UCE-driven gene expression. Phylogenetically conserved motif occurrences of various TFs were also enriched in the noncoding sequence surrounding numerous gene sets corresponding to Gene Ontology categories and tissue-specific gene expression clusters, suggesting involvement in transcriptional regulation of those genes. Altogether, our results indicate that many of the sequences bound by these proteins in vitro, including lower affinity DNA sequences, are likely to be functionally important in vivo. This study not only provides an initial analysis of the potential regulatory associations of 104 mouse TFs, but also presents an approach for the functional analysis of TFs from any other metazoan genome as their DNA binding preferences are determined by PBMs or other technologies.

Mistreatment, discrimination and burn-out in Neurosurgery.

OBJECTIVE: Issues concerning harassment, bullying and discrimination are not unknown to medical specialties and are likely to be present in neurosurgery as well. The aim of this study was to estimate the extent to which neurosurgeons are faced with issues pertaining to this mistreatment. METHODS: A survey consisting of fourteen questions was distributed among members of the Congress of Neurological Surgeons (CNS). The survey consisted of three parts: 1) demographics; 2) exposure to mistreatment; 3) experienced burnout symptoms. RESULTS: In total 503 out of the 5665 approached CNS members filled in a survey (response rate 8.9 %). Respondents consisted for 85.9 % out of neurosurgeons and for 13.9 % out of residents. Overall, 61.4 % of the respondents was a victim of form of abusive behavior, while 47.9 % was a victim of at least one form of discrimination. Most reported sources of these mistreatments were other neurosurgeons or (family of) patients. Overall, 49.9 % of the respondents experienced burnout symptoms. Multivariable logistic regression analysis showed that female respondents had higher odds of being a victim of abuse (OR 2.5, 95 % CI 1.4-4.6). Female respondents (OR 19.8, 95 % CI 8.9-43.9) and ethnic minorities (OR 3.8, 95 % CI 2.3-6.2) had higher odds of being a victim of discrimination. Furthermore, victims of abuse were at higher odds (OR 1.7, 95 % CI 1.1-2.6) of having burnout symptoms. CONCLUSIONS: Mistreatment and experiencing burnout symptoms frequently occurs among neurosurgeons and residents.

Prototyping Trastuzumab Docetaxel Immunoliposomes with a New FCM-Based Method to Quantify Optimal Antibody Density on Nanoparticles.

Developing targeted nanoparticles is a rising strategy to improve drug delivery in oncology. Antibodies are the most commonly used targeting agents. However, determination of their optimal number at the surface remains a challenging issue, mainly due to the difficulties in measuring precisely surface coating levels when prototyping nanoparticles. We developed an original quantitative assay to measure the exact number of coated antibodies per nanoparticle. Using flow cytometry optimized for submicron particle analysis and beads covered with known amounts of human IgG-kappa mimicking various amounts of antibodies, this new method was tested as part of the prototyping of docetaxel liposomes coated with trastuzumab against Her2+ breast cancer. This quantification method allowed to discriminate various batches of immunoliposomes depending on their trastuzumab density on nanoparticle surface (i.e., 330 (Immunoliposome-1), 480 (Immunoliposome-2) and 690 (Immunoliposome-3), p = 0.004, One-way ANOVA). Here we showed that optimal number of grafted antibodies on nanoparticles should be finely tuned and highest density of targeting agent is not necessarily associated with highest efficacy. Overall, this new method should help to better prototype third generation nanoparticles.

[Potentialitis of phosphodiesterase inhibitors in management of patients with chronic heart failure complicating cardiac ischemia].

In spite of the advances in understanding the pathogenesis and approaches to the treatment of chronic heart failure, it remains one of the main causes of the patients invalidism and death, that makes the cardiologists all over the world develop new drugs for the treatment of the disease. The aim of the study was to show possible long-term use of enoximone, a new agent from the group of phosphodiesterase inhibitors, in doses of 75-150 mg a day in traditional combined pharmacotherapy of the patients with chronic heart failure. Thirty eight patients with chronic heart failure III-IV FC (NYHA) and 30% left ventrical efflux function were observed. The dynamics of the disease clinical signs, the heart hemodynamic and morphofunctional parameters, the heart bioelectric activity and the hospitalization frequency were investigated to estimate the therapy efficacy and safety. The use of enoximone in the traditional combined pharmacotherapy, including inhibitors of angiotensin-transforming enzymes, diuretics, beta-adrenoblockers and aldosterone antagonists, resulted in a reliable improvement as for the disease clinical signs, 17.8% increase of the left ventrical efflux function vs. the initial indices, less frequent hospitalization. The enoximone tolerability was good, no side effects requiring discontinuation of the drug use were recorded.

[Dosage of urinary iodures with a crystalline membrane selective electrode]. / Dosage des iodures urinaires à l'aide d'une électrode sélective á membrane cristalline

The fixation of iodine 131 on thyroid is often disturbed by iodine overdose. In order to detect such conditions, the authors propose a method of estimation of urinary iodide based on the use of a selective crystalline membrane electrode.