Oct4 differentially regulates chromatin opening and enhancer transcription in pluripotent stem cells

The transcription factor Oct4 is essential for the maintenance and induction of stem cell pluripotency, but its functional roles are not fully understood. Here, we investigate the functions of Oct4 by depleting and subsequently recovering it in mouse embryonic stem cells (ESCs) and conducting a time-resolved multiomics analysis. Oct4 depletion leads to an immediate loss of its binding to enhancers, accompanied by a decrease in mRNA synthesis from its target genes that are part of the transcriptional network that maintains pluripotency. Gradual decrease of Oct4 binding to enhancers does not immediately change the chromatin accessibility but reduces transcription of enhancers. Conversely, partial recovery of Oct4 expression results in a rapid increase in chromatin accessibility, whereas enhancer transcription does not fully recover. These results indicate different concentration-dependent activities of Oct4. Whereas normal ESC levels of Oct4 are required for transcription of pluripotency enhancers, low levels of Oct4 are sufficient to retain chromatin accessibility, likely together with other factors such as Sox2

Современные тенденции производства беловых товаров

Систематизовано види ділових щоденників і технологи їх виробництва. Показано перспективність цього виду білових товарів.It is systematised kinds of business diaries and technologists of their manufacture. Perspectivity of this kind of the clean goods is shown.Систематизировано виды деловых дневников и технологи их производства. Показана перспективность этого вида беловых товаров

Prevention, diagnosis and management of influenza in CKD patients: Adapted clinical guidelines of the Ukrainian Association of Nephrologists and Kidney Transplant Specialists

Flu is the cause of up to 650,000 deaths worldwide each year. According to CDC USA in 2019-2020 the flu caused 86,000 deaths. Patients with chronic kidney disease (CKD) are at high risk for serious complications and adverse effects of influenza infection. A feature of this year is the possibility of combining influenza with COVID-19, which can significantly complicate treatment and worsen the prognosis of CKD patients. Therefore, prevention and adequate management of influenza in this patient cohort are obvious. In this regard, the Expert Group of the Ukrainian Association of Nephrologists and Kidney Transplant Specialists has created adapted clinical guidelines for the prevention, diagnosis and management of influenza in CKD patients

Morphological peculiarites and functional activity of adipose-derived mesenchimal stem cells during in vitro cultivation conditions

The studies were conducted on 2-3-months-old males of C57BL/6 mice weighing 20–24 g. Obtaining and cultivating of adipose-derived mesenchimal stem cells (AD MSCs) were carried out in a sterile laminar box with compliance of conditions of asepsis and antiseptics. AD MSCs of the 2, 4, 7 and 12 passages were analyzed. Morphometric analysis was performed using a light microscopy. Morphometric parameters such as cell and nucleus area or nuclear-cytoplasmic ratio (NCR) were calculated using the Axiovision light microscope (Carl Zeiss, Germany) and ImageJ 1.45 software. Trypan blue dye used for investigation of the viability of MSC. The morphological characteristics of mesenchymal stem cells from adipose tissue during the process of cultivation changes: at the first passages of cultivation, the cells are spindle-shaped with two, at least three, long long cytoplasmic processes, located bipolar. Near the nucleus the Golgi complex is clearly visible – a sign of active cells. At later passages cells have a small cytoplasmic processes and the bipolar arrangement of processes changes by stellar arrangement. Golgi complex is also clearly visualized. The indicator of the nuclear-cytoplasmic ratio in MSC from adipose tissue is significantly reduced at 7 passage to 0.2189 ± 0.0122 (P < 0.01), and at 12 passage to 0.1111 ± 0.0086 (P < 0.001) compared to the 2 passage. The coefficient of proliferation of MSC from adipose tissue is significantly reduced at 12th passage. The viability of mesenchymal stem cells from adipose tissue with an increasing of a number of passages significantly reduces and at the 12th passage of cultivation reaches 84,67 ± 1,36* (P < 0.05). The content of apoptotic cells that exhibited sensitivity to serum-free significantly increased at 7 and 12 passages and was respectively 21.33 ± 1.36 (P < 0.05) and 23.67 ± 0.97% (P < 0.05)

Three types of gall-bladder motility at men with chronic cholecystitis and pyelonephritis and accompanying their parameters of kidney excretory function as well as metabolism

Background. Previously it has been shown that balneotherapy in the spa Truskavets’ ( Ukra ї na) in men with chronic cholecystitis combined with pyelonephritis causes changes in gall-bladder motility. This is combined with changes in diurese and urinary excretion of metabolites. Individual analysis revealed not only quantitative but also qualitative differences in the postprandial motility of the gall - bladder. The purpose of this study is to identify the parameters of kidney excretory function as well as metabolism under which three types of the gall - bladder motility skills differ significantly from each other. Material and methods. The object of observation were the same ones 22 men aged 24-70 years old, who came to the spa Truskavets’ for the treatment of chronic cholecystitis combined with pyelonephritis in remission. On the tone and motility of gall-bladder judged by its fasting and postprandial volume ( echocamera “Radmir”). In the daily  determined content of oxalate and nitrogen metabolites: creatinine, urea and uric acid, electrolytes: phosphates, chloride, calcium, magnesium, potassium and sodium. Nitrogenous metabolites in plasma of venous blood were determined also. The survey was conducted twice, before and after balneotherapy. Results. The method of cluster analysis has formed three homogeneous cholekinetics groups, namely: normokinesia a s well as hypertonic-hyperkinetic and hyperkinetic-hypertonic dyskinesia. The method of discriminatory analysis revealed 13 parameters of daily urine and three parameters of blood, as well as body weight and height, electrokinetic index and Kerdoe’s index, in the totality of which three types of cholekinetics are clearly delineated. Conclusion. The type of postprandial cholekinetics is naturally associated with a number of parameters of the excretory renal function and metabolism

Percentage expression of neutrophil activation marker in the peripheral blood of patients with dry eye disease plus type 2 diabetes

Background: Dry eye disease (DED) is a disorder of the anterior segment of the eye which is common in patients with type 2 diabetes mellitus (T2DM). It has been reported that symptoms of DED were observed in 60-70% of patients with T2DM. Given that inflammation has a key role in the progression of both T2DM and DED, we believe it is reasonable to study the role of neutrophils in this process. Purpose: To assess the percentage expression of CD15, a neutrophil activation marker, in the peripheral blood of patients with both DED and T2DM. Material and Methods: Forty-six patients (92 eyes; mean age, 54.0 ± 8.0 years) with both DED and T2DM were included in this study. There were 19 (40%) women and 27 (60 %) men. Mean diabetes duration was 8.0 ± 6.6 years. All patients had well-compensated diabetes. In addition to a routine examination of the eye, Schirmer I test and tear film break-up time (TBUT) test, they had their corneal fluorescein staining (CFS) scored using the Oxford schema, severities of dry eye graded according to the DEWS II classification and Ocular Surface Disease Index (OSDI) obtained. An immunohistocytochemical study was employed to assess the expression of CD15, a neutrophil activation marker, in peripheral blood cells of patients. Results: The mean percentage expression of CD15, a neutrophil activation marker, in the peripheral blood, was 46.7% for 14 patients with T2DM plus DED and punctuate keratopathy, and 28.5% for 32 patients with T2DM plus DED and intact cornea (р=0.0001). There was a mild negative correlation between the expression of CD15 and the Schirmer score (r = -0.32; р = 0.032), and between the former and the TBUT score (r = -0.34; р = 0.019)

Особливості показників імунітету у хворих на непроліферативні форми хронічного гломерулонефриту

Мета роботи: визначити особливості HLA-фенотипів та показників цитокінової ланки імунітету з факторами ендотеліальної дисфункції у хворих на непроліферативні (НП) форми хронічного гломерулонефриту (ХГН) з нефротичним синдромом (НС). Матеріал та методи. Вивчали розподіл HLA-антигенів за допомогою мікролімфоцитотоксичного тесту Терасакі у 264 хворих на ХГН, НС (серед яких аналізували дані 96 з НП ГН) та 350 здорових донорів. Морфологічний діагноз був підтверджений методом тонкоігольної нефробіопсії. Для вивчення рівнів цитокінів та факторів ендотеліальної дисфункції в крові 110 пацієнтів використовували імуноферментний аналіз. Результати. Для ХГН, НС відносний ризик високий (RR> 2)за наявності HLA-А23, 24, 28, В8, 38, 41, 44 (у хворих на НП ГН +А30),причинна роль (σ> 0.1)показана для А24, 28, В8 (НП +A19+31+32); протекторами захворювання є В12 та В16 (НП +А9). Виявлені антигени-провокатори різних морфологічних форм ХГН. У пацієнтів з НП ГН виявлено в крові високі рівні як про- (трансформуючий фактор росту альфа (TNF-α), інтерлейкіну 17 (IL-17), моноцитарний хемоатрактантний протеїн 1 (МСР-1)), так і протизапальних (IL-4, трансформуючий фактор росту бета (TGF-β)) цитокінів, факторів ендотеліальної дисфункції (фактор росту ендотелію судин (VEGF), розчинна молекула адгезії судинних клітин (sVCAM-1)) та зниження Е-селектину. РівеньMCP-1 у хворих на проліферативні ГН був вищим, ніж у разі непроліферативних форм. Не виявлено достовірних змін цих показників імунітету в групі хворих на НП ГН після проведеного лікування, за винятком зниження TNF-α (р = 0,001), але позитивний ефект лікування асоціював з достовірним зниженням концентрацій МСР-1 (р = 0,005) і VEGF (р = 0,028). Висновки. Виявлені достовірні асоціаціїHLA з ХГН, НС та його непроліферативними морфологічними формами дозволяють вважати такі антигени в фенотипі пацієнтів та інші показники імунітету ((TNF-α,MCP-1 і VEGF) додатковими прогностичними маркерами

ОЦІНКА ЯКОСТІ ЖИТТЯ ТА ПСИХОЕМОЦІЙНОГО СТАТУСУ У ХВОРИХ НА ХРОНІЧНИЙ ПАНКРЕАТИТ

Researches of a mental and emotional condition of patients with chronic pancreatitis are important. Their quantity is poor. Quality of life and degree of uneasiness at patients with chronic pancreatitis was to estimate the purpose of work. At 68 patients with chronic pancreatitis are estimated on a visual analog scale – intensity of a pain syndrome, according to the questionnaire of GSRS – quality of life, according to Ch.D.Spilbergera-Yu.L.Khanina's questionnaire – existence of alarming frustration. It is established that at patients with chronic pancreatitis are noted a high level of jet and personal uneasiness, than in group of monitoring, deterioration of life, essential changes in the psychoemotional status. Deterioration of their life has a feed forward with expressiveness of painful, diarrheal syndromes.В связи с важностью и недостаточным количеством исследований, отображающих уровень психоэмоционального состояния больных хроническим панкреатитом, целью работы было оценить качество жизни и степень тревожности у больных хроническим панкреатитом. У 68 больных хроническим панкреатитом оценены по визуальной аналоговой шкале – интенсивность болевого синдрома, по вопроснику GSRS – качество жизни, по вопроснику Ч.Д. Спілбергера-Ю.Л. Ханіна – наличие тревожных расстройств. Установлено, что у больных хроническим панкреатитом отмечаются высокий уровень реактивной и личностной тревожности, чем в группе контроля, ухудшение качества жизни, существенные изменения в психоэмоциональном статусе, ухудшение качества жизни имеет прямую  связь с выраженностью болевого, диарейного синдромов.В зв’язку із важливістю і недостатньою кількістю досліджень, що відображають рівень психоемоційного стану хворих на хронічний панкреатит, метою роботи було оцінити якість життя та ступінь тривожності у хворих на ХП. У 68 хворих на ХП оцінені за візуальною аналоговою шкалою – інтенсивність больового синдрому, за опитувальником GSRS – якість життя, за опитувальником Ч.Д.Спілбергера-Ю.Л.Ханіна – наявність тривожних розладів. Встановлено, що у хворих на ХП наявні вищий рівень реактивної та особистісної тривожності, ніж в групі контролю, відмічаються погіршення якості життя, суттєві зміни в психоемоційному статусі, погіршення якості життя має прямий зв'язок з виразністю больового, діарейного синдромів

CDK12 globally stimulates RNA polymerase II transcription elongation and carboxyl-terminal domain phosphorylation

Cyclin-dependent kinase 12 (CDK12) phosphorylates the carboxyl-terminal domain (CTD) of RNA polymerase II (pol II) but its roles in transcription beyond the expression of DNA damage response genes remain unclear. Here, we have used TT-seq and mNET-seq to monitor the direct effects of rapid CDK12 inhibition on transcription activity and CTD phosphorylation in human cells. CDK12 inhibition causes a genome-wide defect in transcription elongation and a global reduction of CTD Ser2 and Ser5 phosphorylation. The elongation defect is explained by the loss of the elongation factors LEO1 and CDC73, part of PAF1 complex, and SPT6 from the newly-elongating pol II. Our results indicate that CDK12 is a general activator of pol II transcription elongation and indicate that it targets both Ser2 and Ser5 residues of the pol II CTD

HLA-PHENOTYPE IN PATIENTS WITH GLOMERULONEPHRITIS WITH VARIOUS MORPHOLOGIC FORMS AND NEPHROTIC SYNDROM

In the work was determined the HLA-phenotype specificities in patients with different morphologic forms of chronic glomerulonephritis and nephrotic syndrome (CGN, NS) to define the additional predictors of a disease course. Materials and methods. There was studied the HLA-antigens distribution in the 264 CGN, NS patients and 350 healthy donors by typing the lymphocytes with the aid ofstandard microlymphocytotoxic test (Terasaki’s test). The diagnosis was confirmed morphologically using the thin needle nephrobiopsy. Results. It is advisable to associate CGN, NS (RR > 2) with antigens HLA- A23, 24, 28; B8, 38, 41, 44 in patients; the causal role (a > 0.1) was determined for A24, 28; B8. In proliferative GN was additionally revealed the etiologic role of B27 known as antigen associated with risk ofautoimmune diseases. In patients with various morphologic forms is advisable the association of some antigens with development of chronic renal failure (CRF) – A30, B41 in FSGS, A10 – MGN; and also hormone resistance (HR) – A19+31+32 in FSGS, B8 – MGN and MC. Conclusion. The revealed reliable associations ofHLA types both with CGN, NS and its separate morphologic forms with the risk of CRF and/or HR allow take into consideration the availability ofsuch antigens in phenotype ofpatients with confirmed by biopsy diagnosis as the additional diagnostic and prognostic markers