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Novel CRISPR-Cas-based genome editing tools made it feasible to introduce a variety of precise genomic modifications in the pig genome, including introducing multiple edits simultaneously, inserting long DNA sequences into specifically targeted loci, and performing nucleotide transitions and transversions. Pigs serve as a vital agricultural resource and animal model in biomedical studies, given their advantages over the other models. Pigs share high similarities to humans regarding body/organ size, anatomy, physiology, and a metabolic profile. The pig genome can be modified to carry the same genetic mutations found in humans to replicate inherited diseases to provide preclinical trials of drugs. Moreover, CRISPR-based modification of pigs antigen profile makes it possible to offer porcine organs for xenotransplantation with minimal transplant rejection responses. This review summarizes recent advances in endonuclease-mediated genome editing tools and research progress of genome-edited pigs as personalized test-systems for preclinical trials and as donors of organs with human-fit antigen profile. Graphical abstract

Atherosclerosis is a systemic autoimmune disease of the arterial wall characterized by chronic inflammation, high blood pressure, oxidative stress, and progressive loss of cell and organ function with aging. An imbalance of mac­rophage polarization is associated with many aging diseases, including atherosclerosis. The polarization toward the pro-inflammatory M1 macrophage is a major promoter of the atheroma formation. It is known that efferocytosis, or ingestion of apoptotic cells, is stimulated by M2 macrophage polarization. A failure of efferocytosis leads to the prolon­gation of chronic pathology in tissue. In addition, fat-laden macrophages contribute to the plague progression by trans­forming into foam cells in response to excess lipid deposition in arteries. In spite of the generally accepted theory that macrophages capture oxidized low-density lipoprotein by phagocytosis and become foam cells, we postulate that the main source of lipid accumulation in foam cells are senescent erythrocytes. Senescent erythrocytes lose their plasticity, which affects the rheological blood properties. It is known that their membrane contains high levels of cholesterol. There is evidence that senescent erythrocytes play a pathogenic role in the atheroma formation after breaking down during flowing through an artery bifurcation. Here we review the current knowledge on the impact of age-associated immune cells and red blood cells modifications on atherogenesis. Graphical abstract

The neurovascular unit (NVU), comprised of endothelial cells, pericytes, smooth muscle cells, astrocytic endfeet and microglia together with neurons, is paramount for the proper function of the central nervous system. The NVU gatekeeps blood-brain barrier (BBB) properties which, as a system, experiences impairment in several neurological and psychiatric diseases, and contributes to pathogenesis. To better understand function and dysfunction at the NVU, isolation and characterization of the NVU is needed. Here, we describe a singular, standardized protocol to enrich and isolate microvessels from archived snap-frozen human and frozen mouse cerebral cortex using mechanical homogenization and centrifugation-separation that preserves the structural integrity and multicellular composition of microvessel fragments. For the first time, microvessels are isolated from postmortem vmPFC tissue and are comprehensively investigated using both RNA sequencing and Liquid Chromatography with tandem mass spectrometry (LC-MS-MS). Both the transcriptome and proteome are elucidated and compared, demonstrating that the isolated brain microvessel is a robust model for the NVU and can be used to generate highly informative datasets in both physiological and disease contexts.

The new radiocarbon dates were obtained from samples of the collections from the excavations by S.N. Zamyatnin in 1955–1957 of the Sosruko Rockshelter in the Elbrus region. The Sosruko Rockshelter is a multi-layered site containing cultural horizons of the Iron Age, Mesolithic and Late Upper Palaeolithic. Clear stratigraphy of the Stone Age layers and representative collections were used to create periodization schemes of the development and change of the lithic industries of the late Pleistocene — early Holocene of the Caucasus. But the lack of radiocarbon dating did not allow determining their absolute age. Three samples of faunal remains of layers M1, M2 and M3 were analyzed. Obtained four radiocarbon AMS dates are in agreement not only with the sequence of deposits in the Rockshelter, but also with the data obtained for similar typological collections of the North-West Caucasus synchronous sites.

Background: It is known that right heart failure contributes significantly to morbidity and mortality among patients with chronic lung diseases. However, there is very little literature describing factors associated with right ventricle (RV) dysfunction among patients with chronic hypersensitivity pneumonitis (HP). Our aim was to investigate factors associated with the RV systolic dysfunction among patients with chronic HP. Material and Methods: We identified 86 patients with chronic HP, who underwent echocardiography, spirometry, plethysmography, diffusing capacity of carbon monoxide (DLCO), bronchoscopy and lung biopsy. Pulmonary high resolution computed tomography (HRCT) was assessed by Kazerooni scale (ground glass and lung fibrosis). Pulse wave velocity (PWV), and BMI were evaluated. RV systolic function was assessed using different methods (tricuspid annular plane systolic excursion (TAPSE), RV myocardial performance index (MPI) and RV systolic excursion velocity by tissue Doppler (S′)). Results: RV systolic dysfunction was found in 40.6% of subjects by TAPSE and 38.4% by RV MPI and S′. All parameters of RV systolic function correlated with total lung capacity (p Conclusions: RV systolic dysfunction is associated with lungs fibrosis, diffusion disturbance and arterial stiffness among patients with chronic HP.

Digital transformation of education poses new challenges in the preparation of future teachers, including teachers of physics. The basis of digital competence of teachers is digital skills, which are manifested in particular in the ability to apply electronic educational resources: training programs, simulators, computer models, etc., as well as a set of such tools - virtual learning environments. The preparation of a future physics teacher for the management of students cognitive activity combines the work of a teacher at the same time in two directions. The first direction is the development by students of the features of the management process of the cognitive activity of schoolchildren. The second direction is the teaching students how to present the elements of content and how to master them in digital form by using ready-made or independently developed electronic educational resources. The implementation of productive technology by the teacher helps to improve the efficiency of preparing the future physics teacher for the management of the cognitive activity of students using digital educational resources. As a result, students embody their knowledge and skills in an educational product. This is a set of teaching and learning materials that is objectively new and significant for their future professional activity. The author of the article presents the experience of developing the content of training future physics teachers to manage learning and cognitive activity in a virtual learning environment which is based on modern ideas about the development of education.

Background: Cardiovascular diseases are the most common comorbidities among patients with chronic lung diseases. Arterial stiffness (AS) is one of the most potent prognostic factors of cardiovascular morbidity and mortality. However, there is very little literature describing the factors associated with the increase of AS among patients with chronic hypersensitivity pneumonitis (HP). Aims and Objectives: were to investigate arterial stiffness and determine factors associated with its increase among patients with HP. Methods: We identified 106 patients with chronic HP, who underwent spirometry, plethysmography, diffusing capacity of carbon monoxide (DLCO), pulmonary high resolution computed tomography (HRCT), bronchoscopy and lung biopsy. AS was measured by pulse wave velocity (PWV). BMI, the duration of HP and the exacerbations frequency were evaluated. Results: The increase of arterial stiffness was found in 71 (67%) of patients. Correlation analysis adjusted for age showed a significant correlation between PWV and DLCO, the extent of lung fibrosis (LF) on HRCT (p Conclusions: The increase of arterial stiffness is associated with the extent of lung fibrosis on HRCT, diffusion disturbance, and frequency of exacerbations among patients with chronic hypersensitivity pneumonitis.

Objectives: To identify risk factors for atrial fibrillation (AF) in patients with chronic obstructive pulmonary disease (COPD). Methods: We examined 94 patients with a 2-3 degree stable COPD (The Global Initiative for Chronic Obstructive Lung Disease 2013) without cardiovascular disease and thyroid dysfunction. Spirometry, daily pulse oximetry, 24-h electrocardiography monitoring and blood pressure monitoring with vascular stiffness (pulse wave velocity in aorta) estimation, and echocardiography were performed. We evaluated the levels of high-sensitivity C-reactive protein (hs-CRP). We found AF paroxysms 2 [1; 4] in 46 COPD patients (COPD and AF group). The other COPD patients (48 in total) comprised a control group. The groups were alike as to age, sex, COPD duration, COPD exacerbation, arterial hypertension (AH) duration and degree. Statistical analysis used multiple correlation analysis, logistic regression. Results: Patients with COPD and AF had more intensive systemic inflammation, lower forced expiratory volume in 1 sec (FEV1) and minimum oxygen saturation in the blood (min%SpO2), more COPD symptoms, higher vascular stiffness. The frequency of paroxysms of AF was correlated with FEV1 (r=-0.348; p=0.013), min%SpO2 (r=-0.356; p=0.011), hs-CRP level (r=0.442; p=0.001), pulmonary artery systolic pressure (PASP) (r=0.530; р Conclusions: According to the results of logistic regression PASP, vascular stiffness, hs-CRP, FEV1 and hypoxemia are risk factors for AF in COPD patients.

Summary Extractive and chromatographic separations were performed on V-1326, a chloroform extract from the bark of Prunus africana (also referred to as Pygeum africanum), which is used to treat the symptoms associated with benign prostate hyperplasia (BPH). The relative amounts of eleven identified constituents in crude V-1326 and in separated fractions were determined using gas chromatographic analysis. The ability of V-1326 and its separated fractions to inhibit ferrous ion-induced stimulation of lipid peroxidation in microsomal preparations from rabbit livers was evaluated. The extract, V-1326, and fractions containing high levels of myristic acid potently inhibited lipid peroxidation.

Partial bladder outlet obstruction results in progressive loss in contractile and specific cellular and subcellular membrane functions. There is evidence that ischemic activation of proteolytic and lipolytic enzymes play a major role in the etiology of bladder dysfunction secondary to partial outlet obstruction. The specific aims of the current study were to determine the fatty acid profiles in normal rabbit bladder smooth muscle and mucosa and to determine the effect of partial outlet obstruction on the distribution and content of free and total fatty acids. Fatty acids were isolated by extraction from obstructed and normal bladder smooth muscle and mucosal homogenates, and samples were analyzed by gas chromatography. All samples contained palmitic, stearic, oleic, linoleic, and arachidonic acids. A 100% increase in total fatty acid concentration was observed in the obstructed bladder muscle tissue relative to normal bladders, although the concentration of total arachidonic acid remained constant in the two groups. Significantly higher levels of free arachidonic acid were observed in the obstructed bladder muscle group compared to the normal group. No changes were observed in fatty acid concentrations or distributions in bladder mucosa. These data show that fatty acid composition is altered as a result of bladder obstruction and support the idea that obstruction increases the activity of lipase activity and/or decreases acyl transferase activity. Neurourol. Urodynam. 18:697-711, 1999.

This study was undertaken to examine in-situ heart function and metabolism during insulin treatment of verapamil-induced cardiogenic shock in awake canines.Twenty mongrel canines were instrumented to monitor myocardial substrate uptakes (glucose, lactate, free fatty acids, oxygen [MVO2]), as well as ventricular (LV) end-systolic elastance (Emax), LV efficiency (LV minute work/MVO2), and Tau. Shock was induced by graded intraportal verapamil infusion followed by randomized assignment to one of 4 treatment groups: saline control (3.0 ml/kg/min, n = 5), epinephrine (5 micrograms/kg/min, n = 5), glucagon (10 micrograms/kg/min, n = 5) or insulin (1000 mU/min, n = 5) with dextrose to clamp arterial [glucose] +/- 10% of basal concentrations.Insulin treatment significantly increased Emax (34 +/- 3 vs. 17 +/- 3 mmHg/mm, saline control), and shortened Tau (9 +/- 3 ms) compared to saline control (42 +/- 5 ms), epinephrine (20 +/- 4 ms) and glucagon (35 +/- 8 ms). With insulin treatment, mechanical efficiency increased to 20,097 +/- 2070 vs. 12,424 +/- 1615 mmHg.mm/ml/O2/100 g in controls. Simultaneously, insulin increased myocardial lactate uptake (35 +/- 2 vs. 17 +/- 4 mumol/min/100 g. saline control), but did not increase glucose uptake. Epinephrine and glucagon decreased mechanical efficiency compared to saline controls, coincident with increased myocardial fatty acid consumption, but without increasing lactate uptake. One dog died early with glucagon treatment before the first death in the saline-treated group.Insulin improves systolic and diastolic heart function during aerobic shock and accelerates in-vivo myocardial lactate oxidation.

Verapamil produces comparatively greater in vivo left ventricular (LV) depression than other calcium channel antagonists produce, possibly because of myocardial metabolic derangements in addition to L-channel antagonism. Therefore, we studied myocardial lipid and carbohydrate usage and the effect of insulin treatment during progressive verapamil toxicity. Verapamil was infused through the portal vein to simulate oral overdose. Eighteen mongrel dogs were instrumented to measure multiple hemodynamic and metabolic parameters. After 1-week recovery, dogs underwent control euglycemic insulin dose-response studies (n = 6) in the conscious state: at 1,000 mU/mm insulin infusion rate, myocardial glucose and lactate extraction increased seven- and threefold, respectively with no change in coronary artery blood flow or ventricular elasticity and end-systole (Ees). In 12 separate dogs, intraportal graded verapamil toxicity was induced in 3 h by increasing the infusion rate hourly: 0.04 -- 0.08 -- 0.1 mg/kg/mm. At the end of hour 3, myocardial extraction of free fatty acids decreased from 33 +/- 4 to 9 +/- 3% (mean +/- SEM, p < 0.05), without significant change in myocardial blood flow or arterial free fatty acid concentration. Verapamil toxicity increased arterial glucose from 3.5 +/- 0.16 to 6.1 +/- 1.1 mM; simultaneously, myocardial glucose extraction doubled, although endogenous insulin concentrations did not increase. Arterial lactate concentrations and net myocardial lactate uptake both increased (p < 0.05 vs basal blue). Ees decreased from 28 +/- 1 mm Hg/mm (basal) to 20 +/- 2 mm Hg/mm (end of hour 3, p

Sprague-Dawley rat hearts were perfused under constant flow conditions, and a balloon was inserted into the left ventricle to measure heart rate (HR) and left ventricular pressures. Left ventricular generated pressure (LVGP) was calculated as peak systolic minus end diastolic pressure. Three substrate groups, pyruvate (5 mm), glucose (15 m m ) and octanoate (0.5 mm), were employed. Oxidative stress was induced by perfusion with tertiary-butyl hydroperoxide (tBHP. 0.35 mm. 12 min) followed by 25 min of perfusion with control buffer. Hearts perfused with pyruvate showed no significant decrease in contractile function following tBHP treatment (HR × LVGP= 17666 ± 585mmHg/min, initial; 16414±2083 post-tBHP treatment). Glucose-perfused hearts had an intermediate decrease in function (19174±828 mmHg/min, initial; 4379±2083 post-tBHP), while octanoateperfused hearts recovered no contractile function. Peak release of LDH was lowest in hearts perfused with pyruvate (115 ± 17 mU/g wet wt/min), intermediate in glucose-perfused hearts (1575+380) and highest in octanoateperfused hearts (3074±499). Thiobarbituric acid reactive substances (TBARS) were unchanged in hearts perfused with pyruvate (16.2 ± 5 nmoles/g wet wt), but increased significantly in glucose-perfused hearts (36.1 ± 1) and in octanoate-perfused hearts (45.5 ±9). Total glutathione levels were unchanged in hearts perfused with pyruvate (753±68 nmoles/g wet wt), but significantly decreased in glucose-perfused hearts (594±68) and in octanoateperfused hearts (445 ± 38) following tBHP-treatment. Pyruvate significantly reduced oxidative injury. In contrast, glucose provided a small reduction in injury while octanoate-perfused hearts had the most severe injury. Measurements of tBHP indicated that pyruvate directly scavenged tBHP from the buffer, while glucose and octanoate were ineffective at removing tBHP.

Myocardial depression from verapamil toxicity may result from alterations in carbohydrate metabolism as well as calcium-channel antagonism. We hypothesized that pharmacologic doses of insulin may be effective in reversing both of these deficits.Randomized, controlled, prospective study.Laboratory of an urban hospital.Thirty mongrel dogs.Thirty mongrel canines were anesthetized with alpha-chloralose. Toxicity was induced by the administration of 0.1 mg/kg/min iv of verapamil, until there was a 50% reduction in mean arterial pressure, for 30 mins (titration), followed by a continuous verapamil infusion of 1 mg/kg/hr. Animals (n = 6 per group) were randomized to the control group (saline only) or to one of four treatment protocols: a) calcium chloride (20 mg/kg), then 0.6 mg/kg/hr; b) hyperinsulinemia-euglycemia (4.0 U/min of recombinant insulin, with arterial glucose concentration clamped to +/- 10 mg/dL [+/- 0.5 mmol/L] of the basal value); c) epinephrine, with a starting rate of 1.0 microgram/kg/min, titrated to maintain left ventricular pressure at basal values; or d) glucagon, a 0.2-mg/kg bolus, followed by a 150-microgram/kg/hr infusion. Animals were monitored until death or 240 mins; infusate volumes were held constant for all groups.During verapamil titration, the myocardial respiratory quotient increased from 0.84 +/- 0.05 to 1.07 +/- 0.11 (p.05, paired t-test) and myocardial glucose uptake doubled, despite a reduction in cardiac work (p.05, paired t-test). Net myocardial lactate uptake also increased significantly, excluding myocardial ischemia. In controls, this trend continued, indicating preferential carbohydrate metabolism during untreated verapamil toxicity. Despite hyperglycemia, the plasma insulin concentration was not significantly different in controls (basal value 11 +/- 2 vs. 39 +/- 21 microU/mL at 30 mins). Hyperinsulinemia-euglycemia increased both myocardial glucose and lactate uptake five-fold, and significantly increased the ratio of myocardial oxygen delivery/work, along with superior improvements in maximal left ventricular elastance at end systole compared with other treatments (p.05 vs. other treatments, contrast analysis).Verapamil toxicity renders the heart dependent on carbohydrate metabolism. Inasmuch as the positive inotropic effects of all treatments were coincident with increased indices of myocardial carbohydrate uptake, adequate treatment of verapamil toxicity appeared to require maximal myocardial carbohydrate utilization. Hyperinsulinemia-euglycemia allows larger increases in myocardial carbohydrate metabolism and myocardial contractility than calcium chloride, epinephrine, or glucagon, resulting in improved survival rates during severe verapamil toxicity.