Selective bypass of a lagging strand roadblock by the eukaryotic replicative DNA helicase.

The eukaryotic replicative DNA helicase, CMG, unwinds DNA by an unknown mechanism. In some models, CMG encircles and translocates along one strand of DNA while excluding the other strand. In others, CMG encircles and translocates along duplex DNA. To distinguish between these models, replisomes were confronted with strand-specific DNA roadblocks in Xenopus egg extracts. An ssDNA translocase should stall at an obstruction on the translocation strand but not the excluded strand, whereas a dsDNA translocase should stall at obstructions on either strand. We found that replisomes bypass large roadblocks on the lagging strand template much more readily than on the leading strand template. Our results indicate that CMG is a 3' to 5' ssDNA translocase, consistent with unwinding via "steric exclusion." Given that MCM2-7 encircles dsDNA in G1, the data imply that formation of CMG in S phase involves remodeling of MCM2-7 from a dsDNA to a ssDNA binding mode.

5†µm CW Ce3+-doped chalcogenide glass fiber laser with 17% slope efficiency.

Efficient room temperature mid-infrared laser action in a Ce3+-doped chalcogenide fiber was demonstrated. The fiber had a doped selenide glass core in an undoped sulfide glass cladding. The pump source was a CW Fe2+:ZnSe laser emitting at 4.14 µm. The optimized fiber length allowed obtaining up to 7 mW of 5.06 µm output with 17% slope efficiency at room temperature.

Charge carrier transport in PbS films doped with iodine.

The results of the measurements of electrical and Hall resistivities on polycrystalline PbS films doped with iodine obtained through hydrochemical deposition are presented. The analysis of the temperature dependence of resistivity points out the crossover from the hopping mechanism due to thermal delocalization in the impurity band to the variable range hopping mechanism. The increase in the iodine content in the films leads to an increase in the impurity ionization energy. It has been established that the temperature dependence of resistivity over a wide temperature range obeys the inverse Arrhenius law, which is characteristic of disordered polycrystalline films with different sizes and orientations of crystallites relative to the substrate, as confirmed by AFM topography, Raman spectra and X-ray diffraction measurements. We found that the type of charge carrier changes from electrons to holes with an increase in the iodine content. Additionally, for a wide range of iodine doping, the concentration of charge carriers is low, indicating the possible occurrence of a self-compensation mechanism due to the formation of impurity defects.

"A National Review of Crowdfunding for Plagiocephaly Helmets".

OBJECTIVE: Custom prescription helmets for plagiocephaly may be a significant financial burden for families, especially when not covered by insurance. This study aims to identify factors that influence the success of crowdsourcing campaigns for this therapy. DESIGN: GoFundMe campaigns were collected by searching terms such as "plagiocephaly" and "baby helmet." Two reviewers analyzed each campaign for variables, including demographic data, story elements, and photo characteristics. Univariate logistic regression was used to determine each variable's impact on success, defined as attaining ≥75% of a campaign goal and significance of p ≤ 0.05. RESULTS: Campaign data from 2011 to 2022 were analyzed. Initial search yielded 1464 campaigns; among these 413 met final inclusion criteria. On average, campaigns raised $2005 (range: $0-$7799) and requested $3151 (range: $160-$30,000). In total, 228 (54%) achieved success, 167 (40%) met their goal, and 35 (8%) raised no funds. A total of $828,256 was raised from the requested $1,301,317. The average reported age was six months (range: 2-17 m). Significant factors associated with success were military affiliation, providing multiple images, including a quoted cost, providing campaign updates, indicating a sense of urgency, diagnosis of torticollis, and mentioning possible complications without treatment. Raising additional funds for therapy, multiple helmets, and unrelated medical costs negatively impacted success. Racial disparities were observed between campaigns. Additionally, regional differences were noted between campaigns. CONCLUSIONS: Crowdsourcing can be a successful endeavor for some families experiencing financial hardships from helmet therapy. This study highlights current gaps within healthcare coverage for helmet treatment and identifies various factors influencing crowdfunding campaigns.

The Study of the Effectiveness of Ethylmethylhydroxypyridine Succinate in Acute Alcohol Intoxication.

The effectiveness of ethylmethylhydroxypyridine succinate (EMHPS) in acute alcohol intoxication was tested in a study on SPF male outbred ICR mice. Ethanol (concentration 40%) was administered to animals once intraperitoneally at a dose of 4 g/kg. Control animals were injected with saline in an equivalent volume. In 15 min after the administration of alcohol, the animals were injected intravenously or intramuscularly with EMHPS at a dose of 50 or 100 mg/kg or with saline via the same route in an equivalent volume. Animal behavior was tested 3 and 24 h later after administration of the substances. After 3 and 24 h, mice in the pathological control groups developed semiptosis, the gait and the turning over reflex were impaired, the strength of the hind limbs decreased and the distance between the hind limbs increased when landing; in the open-field test, the latency of the first movement increased, and the number of rearing postures decreased. Intravenous and intramuscular administration of EMHPS in doses of 50 and 100 mg/kg had a pronounced antitoxic and neuroprotective effect in acute alcohol intoxication: all studied parameters did not differ significantly from the control.

Analysis of Immunobiological Properties of Recombinant Streptococcus pneumoniae Pneumolysin.

We compared the immunogenicity of recombinant S. pneumoniae pneumolysin (rPly) when administered with and without Al(OH)3 adjuvant, and evaluated the protective properties of recombinant protein in the active defense experiment. It was shown that double immunization with rPly+Al(OH)3 increases the levels of IgG antibodies in comparison with the control (p<0.01), while triple immunization results in a more significant increase in IgG antibody levels (p<0.001). Double immunization with rPly without Al(OH)3 does not induce a significant increase in antibody levels in comparison with the control, while triple immunization results in a slight but significant increase in antibody levels (p<0.05). The active defense test proved the protective activity of rPly against S. pneumoniae serotype 3 at intranasal infection.

Gastroprotective Effect of GABA in Metabolic Stress.

We studied the effect of enteral administration of GABA on the gastric mucosa in male Wistar rats (n=47) with modeled metabolic stress (food deprivation for 9 days with free access to water). The relative weights of the adrenal glands and thymus were determined, and histological examination of the stomach was performed. In control rats, modeling the metabolic stress was accompanied by the development of erosive damage to the gastric mucosa related to blood supply disturbances. Administration of GABA prevented erosions and exhibited a pronounced gastroprotective effect. Thus, administration of GABA can be a promising method for the prevention and treatment of erosive gastric lesions associated with metabolic stress.

Suprastin (Chloropyramine) Causes Proarrhythmic Deterioration of Excitation Conduction, Depolarization and Potentiates Adrenergic Automaticity in the Pulmonary Veins Myocardium.

A number of pharmacological drugs have side effects that contribute to the occurrence of atrial fibrillation, the most common type of cardiac rhythm disorders. The clinical use of antihistamines is widespread; however, information regarding their anti- and/or proarrhythmic effects is contradictory. In this work, we studied the effects and mechanisms of the potential proarrhythmic action of the first-generation antihistamine chloropyramine (Suprastin) in the atrial myocardium and pulmonary vein (PV) myocardial tissue. In PV, chloropyramine caused depolarization of the resting potential and led to reduction of excitation wave conduction. These effects are likely due to suppression of the inward rectifier potassium current (IK1). In presence of epinephrine, chloropyramine induced spontaneous automaticity in the PV and could not be suppressed by atrial pacing. Chloropyramine change functional characteristics of PV and contribute to occurrence of atrial fibrillation. It should be noted that chloropyramine does not provoke atrial tachyarrhythmias, but create conditions for their occurrence during physical exercise and sympathetic stimulation.

Redox-Active Compounds in the Therapy of Drug-Resistant Murine Leukemia P388 Strains.

The efficiency of combinations of cytostatics cisplatin and adriamycin with antioxidant sodium 3-(3'-tert-butyl-4-hydroxyphenyl)propyl thiosulfate (TS-13), and nitric oxide (NO) donor NaNO2 was evaluated on two drug-resistant strains of leukemia P388 with changed redox-status of cells. Simultaneous use of both NO donor and TS-13 in combinations with the cytostatics did not increase the efficiency of therapy. In addition, antioxidant activity of TS-13, NaNO2, and their combinations was studied by the method of luminol-dependent chemiluminescence on the model systems with the use of the homogenized cells of sensitive strain and two drug-resistant strains of leukemia P388. It was shown that TS-13 and NO donor produced opposite effects: TS-13 decreased, while NO donor increased the content of free radicals in the model system. Combinations of antioxidant TS-13 and NO donor should be used with consideration for the redox-status of tumor treated.

Automated Determination of Morphometric Parameters of the Human Radius Based on the Results of Its Computed Tomography.

A fundamentally new software method for automated measurement of morphometric parameters of computed tomographic image of the human radius has been designed. The parameters obtained by manual morphometry of computed tomograms using the RadiAnt DICOM Viewer software were compared with the results of the software product developed by us. The relative error in the measurement of morphometric parameters was about 10%. In the future, the presented software method of automated determination of morphometric parameters of the bone will enable modeling of individual prosthesis of the head of the radius based on the results of computed tomograms with consideration of the structural features of the human radius.

Mutagenic Effect during Combined Exposure to Ionizing and Non-Ionizing Electromagnetic Radiation.

Using the method of dominant lethal mutations, we assessed the frequency of the death of Drosophila melanogaster embryos under combined exposure to ionizing γ-radiation and non-ionizing pulsed magnetic field at various doses and modes of exposure. Mutagenic effect of combined exposure is antagonistic in nature. The antagonism is more pronounced when the following mode of exposure was used: exposure to non-ionizing pulsed magnetic field for 5 h followed by exposure to γ-radiation at doses of 3, 10, and 60 Gy. In case of reverse sequence of exposures, the antagonistic effect was statistically significant after exposure to γ-radiation at doses of 3 and 10 Gy, whereas at a dose of 20 Gy, a synergistic interaction was noted.

Assessment of the Sensitivity of Foodborne Cronobacter spp. Strains and Other Enterobacteria to Temperature Stresses and Chlorine-Containing Biocides.

We examined the effects of elevated temperatures and biocides on survivability of food isolates of Cronobacter spp. (C. sakazakii) and concomitant enterobacteriaceae obtained in microbiological control of infant nutrition products. Increased resistance of certain strains of Cronobacter, Enterobacter cloacae, and Pantoea spp. to thermal processing was revealed. Salmonella, Pantoea, and Cronobacter bacteria were least sensitive to antimicrobial action of chlorine-containing agents. The above properties varied in the strains of the same species. Specifically, only two of three examined isolates of Cronobacter spp. demonstrated lower sensitivity to heat in comparison with the enterobacterial test-cultures of other species.

Proinflammatory Activation of Monocytes in Patients with Immunoinflammatory Rheumatic Diseases.

The pathogenesis of immunoinflammatory rheumatic diseases (IRDs) is based on chronic inflammation, one of the key mechanisms of which may be abnormal activation of macrophages, leading to further disruption of the immune system. OBJECTIVE: . The objective of this study was to evaluate the proinflammatory activation of circulating monocytes in patients with IRDs. MATERIALS AND METHODS: . The study involved 149 participants (53 patients with rheumatoid arthritis (RA), 45 patients with systemic lupus erythematosus (SLE), 34 patients with systemic scleroderma (SSc), and 17 participants without IRDs) 30 to 65 years old. Basal and lipopolysaccharide (LPS)-stimulated secretion of monocytes was studied in a primary culture of monocytes obtained from blood by immunomagnetic separation. Quantitative assessment of the cytokines tumor necrosis factor α (TNF-α), interleukin 1ß (IL-1ß), as well as the chemokine monocyte chemoattractant protein-1 (MCP-1) was carried out in the culture fluid by ELISA. Proinflammatory activation of monocytes was calculated as the ratio of LPS-stimulated and basal secretions. RESULTS: . It was shown that the basal secretion of all studied cytokines was significantly increased in all groups of patients with IRDs, except for the secretion of IL-1ß in the SLE group, compared to the control. LPS-stimulated secretion of TNF-α was increased and MCP-1 was decreased in patients with IRDs compared to the control group; LPS-stimulated IL-1ß secretion only in the SSc group significantly differed from the control group. In the RA group, monocyte activation was reduced for all cytokines compared to the control; in the SLE group, for TNF-α and MCP-1; in the SSc group, for MCP-1. CONCLUSIONS: . The decrease in proinflammatory activation of monocytes in patients with IRDs is due to a high level of basal secretion of cytokines, which can lead to disruption of the adequate immune response in these diseases and is an important link in the pathogenesis of chronic inflammation.

Complete Mitochondrial Genome of Clupeonella abrau (Maliatsky, 1930) (Clupeiformes), an Endemic Freshwater Fish from the Lake Abrau (Russia).

The Abrau sprat (tyulka or sardelka) Clupeonella abrau (Maliatsky, 1930) is an endemic fish of the Lake Abrau (Krasnodar Krai, Russia). The full C. abrau mitochondrial genome (16 650 bp) showed a gene arrangement conserved in Clupeidae and 98.8% similarity with the mitochondrial genome of the related species Black and Caspian Sea sprat C. cultriventris from the Black Sea. The COX1 gene sequence was additionally studied in a museum specimen collected in the Lake Abrau in 1938. Variability in modern Abrau sprat COX1 gene locus was estimated at approximately 0.15%, the difference between C. abrau and C. cultriventris was 1.2%, and the difference between the museum and modern C. abrau specimens from the Lake Abrau was 0.92%. The study confirmed that the Abrau sprat is present in the fish community and is capable of reproducing in the lake. Various scenarios were proposed to explain colonization of the Lake Abrau by C. abrau.

[Amplifiation of the c-MYC gene in acinar prostate adenocarcinoma. Morphogenic comparisons]. / Amplifikatsiya gena c-MYC pri atsinarnoi adenokartsinome prostaty. Morfogeneticheskie sopostavleniya.

OBJECTIVE: The purpose of this work was to evaluate c-MYC gene amplification in the substrate of prostate acinar adenocarcinoma at various Gleason scores and various stages of the disease, taking into account the morphological characteristics of the tumor. MATERIAL AND METHODS: The number of cases in the study was 82, including the control group - 12 cases. Morphological assessment included: determination of the total Gleason score, grading group, assessment of lymphovascular/perineural invasion, and architectural characteristics of the tumor. Gene amplification was assessed by FISH using the c-MYC (8q24)/SE8 probe. RESULTS: In all cases of the study group, amplification of the c-MYC gene was detected in the tumor, with a significant difference from the control group (p<0.05). When assessing cases with 4-6 fold copies of the gene, significant differences were established between patients with stages II and III of the disease and stage IV (10.0 and 13.5 versus 30.0) (p<0.05). Cluster amplification of the c-MYC gene was detected with equal frequency in groups of patients with stages III and IV of the disease, while in stage II of the disease, the event almost did not occur (p<0.05). A significant increase in the level of c-MYC gene amplification was found in groups with advanced stages of the disease (p<0.02). Non-cluster amplification significantly distinguishes T4M0 and T4M1 stage patients from the rest with a significant increase in the score (p<0.05). In the metastatic stage of the disease, there was an increase c-MYC gene amplification compared to the non-metastatic stage (p<0.02). The copy number of the c-MYC gene was significantly higher in cases with perineural and lymphovascular invasion, as well as in cases of cribriform tumor organization (p<0.05). CONCLUSION: Amplification of the c-MYC gene in prostate tumor cells is associated with advanced stages of the disease (T4M0 and T4M1) with an increase in the copy number of the gene during the metastatic stage of the process. It was found that increased amplification of the c-MYC gene distinguishes groups of patients whose tumors exhibit perineural and lymphovascular invasion, as well as a cribriform pattern of tumor organization.

Chronic Heart Failure in Patients Hospitalized in 2002 and 2021: Comparative Analysis of Prevalence, Clinical Course and Drug Therapy.

AIM: Comparative analysis of the prevalence of chronic heart failure (CHF), clinical and medical history data, and drug therapy of patients admitted to a cardiology hospital in 2002 and 2021. MATERIAL AND METHODS: The study analyzed the medical records of patients with a confirmed diagnosis of CHF who were admitted in 2002 (n=210) and 2021 (n=381) to a specialized cardiology hospital. RESULTS: According to medical records of 2021, the proportion of patients with a confirmed diagnosis of CHF (87.6%) in the cohort of patients admitted to a cardiology hospital was twice as high as in 2002 (46.4%; p<0.001). The majority of patients with CHF in the study sample were patients with preserved left ventricular ejection fraction (HFpEF). The proportion of such patients significantly increased to reach 75.9% in 2021 compared to 58.6% in 2002 (p<0.001). At the same time, the number of severe forms of CHF (NYHA functional class (FC) IV) decreased by 10% and was 13.2% in 2002 and 1.3% in 2021 (p<0.001). In the majority of patients, ischemic heart disease (98.1 and 91.1% in 2002 and 2021, respectively, p<0.001) and hypertension (80.5 and 98.2%, respectively, p<0.001) were diagnosed as the cause for CHF. Furthermore, the incidence of comorbidity increased significantly: atrial fibrillation was detected in 12.3% of patients in 2002 and 26.4% in 2021 (p < 0.001); type 2 diabetes mellitus, in 14.3 and 32% of patients (p <0.001); and obesity, in 33.3 and 43.7% of patients, respectively (p=0.018). The frequency of using the major groups of drugs increased during the analyzed period: renin-angiotensin-aldosterone system blockers were administered to 71.9% of patients in 2002 and to 87.7% in 2021 (p<0.001); beta-blockers were administered to 53.3 and 82.4% of patients (p<0.001); and mineralocorticoid receptor antagonists, to 1.9 and 18.6% of patients, respectively (p=0.004). CONCLUSION: In 2021, the proportion of patients with a confirmed diagnosis of CHF in the patient cohort admitted to a cardiology hospital was twice as high as in 2002; the phenotype with preserved left ventricular ejection fraction predominated in the CHF structure. During the analyzed twenty-year period, the prevalence of comorbidities increased among CHF patients. The prescription frequency of pathogenetic evidence-based therapy has significantly increased by 2021, however, it remains insufficient even in patients with CHF with reduced left ventricular ejection fraction.

[Features of the Reperfusion Therapy for ST-Segment Elevation Myocardial Infarction According to the Russian Registry of Acute Myocardial Infarction - REGION-IM].

AIM: Based on data from the Russian REGION-IM registry, to study the features of reperfusion therapy in patients with ST-segment elevation myocardial infarction (STEMI) in real-life clinical practice. MATERIAL AND METHODS: REGION-IM is a multicenter prospective observational study. The observational period is divided into 3 stages: during the stay in the hospital and at 6 and 12 months after inclusion in the registry. The patient's records contain demographic and history data; information about the present case of MI, including the time of the first symptom onset, first contact with medical personnel, and admission to the hospital; coronary angiography (CAG) data, percutaneous coronary intervention (PCI) data, and information about the thrombolytic therapy (TLT). RESULTS: Reperfusion therapy was performed in 88.9 % of patients with STEMI. Primary PCI (pPCI) was performed in 60.6 % of patients. The median time from the onset of symptoms to pPCI was 315 minutes [195; 720]. The median time from ECG to pPCI was 110 minutes [84;150]. Isolated TLT was performed in 7.4 %, pharmaco-invasive treatment tactics were used only in 20.9 % of cases. The median time from ECG to TLT (prehospital and in-hospital) was 30 minutes [10; 59], whereas the median time from ECG to prehospital TLT was 18 minutes [10; 39], and in 63 % of patients, TLT was performed more than 10 minutes after diagnosis. PCI followed TLT in 73 % of patients. CONCLUSION: The frequency of reperfusion therapy for STEMI in the Russian Federation has increased considerably in recent years. The high frequency of pPCI is noteworthy, but the timing of pPCI does not always comply with clinical guidelines. The results of this registry confirm the high demand for pharmaco-invasive strategies in real-life clinical practice. Taking into account geographical and logistical features, implementing timely myocardial reperfusion requires prehospital TLT. However, the TLT frequency in the Russian Federation is still insufficient despite its proven maximum effectiveness in the shortest possible time from the detection of acute MI.

[Fluorescence lymphoscopy with sentinel lymph node biopsy in research of lymphatic flow in early gastric cancer]. / Vozmozhnosti flyuorestsentnoi limfoskopii s biopsiei signal'nykh limfaticheskikh uzlov v izuchenii kharaktera limfoottoka pri rannem rake zheludka.

OBJECTIVE: To analyze the incidence and risk factors of regional lymph node metastatic lesions, as well as patterns of lymphatic drainage in early gastric cancer using indocyanine green (ICG). MATERIAL AND METHODS: A prospective single-center non-randomized study included 67 patients with early gastric cancer between 2016 and 2019. All patients underwent endoscopic dissection or surgical resection with sentinel lymph node biopsy. RESULTS: Regional lymph node metastatic lesions were detected in 12 (20.2%) out of 59 patients with examined lymph nodes. Of these, 11 out of 12 patients had sentinel lymph node lesion, and only 1 patient had metastasis outside this lymph node. The proposed technique for sentinel lymph node biopsy demonstrated high sensitivity, specificity and accuracy (93.2%, 100% and 98.3%, respectively). Risk factors for metastasis of early gastric cancer to regional lymph nodes were high grade adenocarcinoma (p=0.038) and tumor of anterior gastric wall (p=0.013). All patients with tumors of the upper third of the stomach had sentinel lymph node in the left gastric artery basin. Lymph flow was multidirectional in patients with tumors of the middle third of the stomach. In tumors of the lower third of the stomach, sentinel lymph node was located in the right gastroepiploic artery (50%) and left gastric artery basins (36.84%) (p=0.002). Sentinel lymph node was located in the left gastric artery basin in 84% of patients with tumors of the lesser curvature (p<0.001). Lymphatic drainage was multidirectional if tumors were localized on anterior and posterior walls, as well as on the greater curvature. CONCLUSION: The proposed technique for sentinel lymph node biopsy demonstrated high sensitivity, specificity and accuracy (93.2%, 100% and 98.3%, respectively). ICG is valuable to identify the first lymph node in lymphatic drainage pathway in 83.6% of cases that makes it possible to assess the regional lymphatic collector. In addition, sentinel lymph node biopsy with ICG contributes to analysis of lymphatic drainage in early gastric cancer. The last one has both scientific and practical significance.

[Diagnostic criteria for prolonged and chronic disturbances of consciousness after aneurysmal subarachnoid hemorrhages]. / Diagnosticheskie kriterii prodlennykh i khronicheskikh narushenii soznaniya posle anevrizmaticheskikh subarakhnoidal'nykh krovoizliyanii.

BACKGROUND: In recent years, prolonged states of impaired consciousness became widespread among patients with aneurysmal subarachnoid hemorrhage. Treatment and maintenance of vital functions in such patients represent a complex medical, economic and social problem. In this regard, searching for the causes of prolonged states of impaired consciousness and predicting the outcomes are important. OBJECTIVE: To analyze available literature data on prevention and treatment of prolonged states of impaired consciousness after aneurysmal subarachnoid hemorrhage. RESULTS: We reviewed the PubMed database using the keywords «unresponsive wakefulness syndrome¼, «persistent vegetative state2, «minimal consciousness state¼ and «outcome of subarachnoid hemorrhage¼. Only 4 reports devoted to the causes and treatment outcomes in patients with prolonged impairment of consciousness after aneurysmal subarachnoid hemorrhage were found. At the same time, patients with aneurysmal subarachnoid hemorrhage comprise up to 11% among all cases of prolonged states of impaired consciousness. Examination, management and treatment of patients with prolonged impairment of consciousness after aneurysmal subarachnoid hemorrhage are carried out according to general principles without taking into account specific etiological and pathogenetic factors. CONCLUSION: Increased number of patients with prolonged impairment of consciousness after aneurysmal subarachnoid hemorrhage necessitates analysis of etiopathogenesis and outcomes of these disorders based on modern clinical, instrumental and laboratory assessment of the brain.

[Long-term result of aortic root remodeling using the Florida sleeve technique in a patient with connective tissue dysplasia and aortic root aneurysm without aortic insufficiency]. / Otdalennyi rezul'tat ekzoprotezirovaniya kornya aorty po metodike Florida Sleeve u bol'nogo s soedinitel'notkannoi displaziei i anevrizmoi kornya aorty bez aortal'noi nedostatochnosti.

Aortic root remodeling using the Florida sleeve technique was proposed as a simpler alternative to classical reconstructive interventions in patients with aortic root aneurysm without aortic insufficiency. We present a 10-year result of valve-sparing surgical treatment using the Florida sleeve technique in a patient with connective tissue dysplasia and aortic root aneurysm.