Electrophilic and Drug-Induced Stimulation of NOTCH3 N-terminal Fragment Oligomerization in Cerebrovascular Pathology.

Small vessel disease is a prevalent age-related condition linked to increased risk of dementia and stroke. We investigate the most commonly inherited form, CADASIL, caused by cysteine-involving mutations in NOTCH3. Recent studies highlight accumulation of NOTCH3 N-terminal fragmentation product (NTF) in disease. In vitro, NTF is capable of both spontaneous and catecholamine-enhanced cysteine-mediated oligomerization. Despite well-characterized genetic influence on CADASIL, environmental effects, including medication usage, on disease remain unclear. We studied effects of assorted electrophilic compounds and drugs on NTF oligomerization by SDS-PAGE and dynamic light scattering. We then examined direct proton pump inhibitor-NTF binding with antibodies designed against proton pump inhibitor-labeled proteins and mass spectrometry. Finally, we used monoclonal NTF antibodies with Proximity Ligation Assay to identify NTF oligomers in 3 CADASIL and 2 age-matched control brains. We identified enhancement of NTF oligomerization by two electrophilic cysteine-modifying compounds, N-ethylmaleimide and iodoacetamide, and an electrophilic compound capable of oxidizing cysteines, ferric chloride. Electrophilic clinical drugs (fenoldopam, omeprazole, tenatoprazole, lansoprazole, and rabeprazole) also promoted oligomerization, and we identified direct omeprazole-NTF and tenatoprazole-NTF complexes. Additionally, we provide novel evidence of NTF multimers in human CADASIL brains. A broad array of electrophilic chemicals, including clinically relevant drugs, influences oligomerization of a pathological CADASIL protein, providing mechanistic insight into disease protein oligomerization. We posit that environmental influences, which may include usage of electrophilic drugs, may affect CADASIL presentations.

Investigation of Taurine Derivative Magnesium-Bis-(2-Aminoethanesulfonic)-Butadioateon Alleviation of Neurological Defects in Simulated Hemorrhagic Stroke in Rats.

Stroke or ischemia is caused by a blockage in a specific blood vessel that partially or completely reduces the blood flow to the brain. Nutritional factors such as antioxidants and healthy eating patterns are important variables in preventing stroke. Molecular composition properties such as molecular binding and screening can be used to evaluate the specific activity and morphological changes. The present study aimed to evaluate the effectiveness of pharmacological correction of the consequences of a hemorrhagic stroke in rats with a new derivative of taurine magnesium-bis-(2-aminoethanesulfonic)-butadioate. The animals (n=170) were divided into four groups as follows: 1) control group (n=20), 2) group 2 suffered a hemorrhagic stroke without pharmacological correction (n=50), 3) group 3 (n=50) underwent simulation of hemorrhagic stroke received Taurine at the dose of 50 mg/kg, 4) Group 4 underwent simulation of hemorrhagic stroke with correction of hemorrhagic stroke with magnesium-bis-(2-aminoethanesulfonic)-butadioate at the dose of 150 mg/kg (LKHT 3-17) (n=50). Hemorrhagic stroke was induced by transfusing autologous blood into the parietal lobe of the right hemisphere of the brain. Lethality, neurological status, locomotor, and exploratory behavior, as well as the morphological pattern of the brain damage, were assessed on the 1st, 3rd, and 7th days after the pathology simulation. Neurological deficit was determined in animals by the McGrow stroke index scale. The locomotor and exploratory behavior was evaluated using the Acti-track software and hardware complex. Two criteria were considered when assessing morphological changes in the brain: the average thickness of the cerebral cortex (in micrometers) and the number of neurons without degenerative changes. LKHT 3-17 (150 mg/kg) and taurine (50 mg/kg) reduced lethality by 1.7 and 1.36 times, respectively, on the 3rd day after stroke compared to that of the control (p<0.05). In parallel, a neurological deficit was effectively corrected LKHT 3-17 and taurine to 5.3±0.8 and 6.5±0.9, respectively, on the 1st day in contrast to the control of 8.1±0.7 points. The locomotor and exploratory behavior was significantly different on the 7th day and was accompanied by a significant increase in total activity under the influence of LKHT 3-17 to 491 conventional units (CU) compared to the control of 110 conventional units. On the 1st day, the thickness of the cortex was 1943.7±44.08 µm, and 1491.0±38.61 µm in the control and LKHT 3-17 groups, respectively. The number of neurons without neurodegenerative changes prevailed in LKHT 3-17 group (18.7±4.32), and the lowest number was observed in the group without pharmacological correction of the pathology (14.3±3.78). The taurine derivative magnesium-bis-(2-aminoethanesulfonic)-butadioate, which is a combination of the amino acid, magnesium ion, and succinic acid, decreases the neurological deficits, lethality, and enhances the locomotor and exploratory behavior in experimental hemorrhagic stroke in rats. The effect of the studied medication on the dynamics of molecular pathophysiological mechanisms occurring in the cell requires additional research.

Infectious Plant Diseases: Etiology, Current Status, Problems and Prospects in Plant Protection.

In recent years, there has been an increase in the number of diseases caused by bacterial, fungal, and viral infections. Infections affect plants at different stages of agricultural production. Depending on weather conditions and the phytosanitary condition of crops, the prevalence of diseases can reach 70-80% of the total plant population, and the yield can decrease in some cases down to 80-98%. Plants have innate cellular immunity, but specific phytopathogens have an ability to evade that immunity. This article examined phytopathogens of viral, fungal, and bacterial nature and explored the concepts of modern plant protection, methods of chemical, biological, and agrotechnical control, as well as modern methods used for identifying phytopathogens.

[Endovascular surgery in treatment of patients with acute impairment of mesenteric circulation in the stage of intestinal ischaemia]. / Éndovaskuliarnaia khirurgiia pri lechenii bol'nykh s ostrym narusheniem mezenterial'nogo krovoobrashcheniia v stadii ishemii kishki.

Analysed herein are the results of endovascular revascularization of the superior mesenteric artery (SMA). The study included a total of 18 patients with acute impairment of mesenteric circulation in the stage of intestinal ischaemia. The patients underwent multispiral computed tomography (MSCT) and once impairments of blood flow in the SMA were revealed, we performed endovascular revascularization of the artery. Patency of the SMA was assessed by repeat contrast-enhanced MSCT. Thirteen (72.2%) patients were found to have occlusion in the system of the SMA and five (27.8%) were diagnosed as having significant stenoses of the SMA. All 13 patients with occlusion of the SMA underwent vacuum thrombextraction followed by transluminal balloon angioplasty (TBA). Of these, thrombotic masses were obtained in 11 (84.6%) patients. Stents were implanted in 3 cases wherein TBA turned out inefficient. The patients with haemodynamically significant stenoses of the SMA were subjected to TBA followed by stent implantation. Good roentgenoendovascular results of restoration of blood flow through the SMA and its branches were obtained in 83.4% of cases. Assessing blood parameters and patients' condition severity revealed positive dynamics or a tendency towards improvement. The mortality rate amounted to 16.6%. A conclusion drawn is that this technique is efficient and appropriate for SMA lesions of any localization.

Mitochondrial DNA variants in Bulgarian patients affected by multiple sclerosis.

The occurrence of multiple sclerosis (MS) in subjects clustering to a particular mitochondrial DNA (mtDNA) haplogroup/haplotype or carrying mtDNA mutations associated with Leber's hereditary optic neuropathy (LHON) has suggested that mitochondrial genome may contribute to susceptibility to MS. In the present study, 58 unrelated Bulgarian patients with relapsing remitting form of MS and 104 randomly selected healthy individuals were analysed for the presence of 14 mtDNA polymorphisms determining major European haplogroups as well as three (4216, 14 798, 13 708) secondary LHON mutations. Restriction enzyme analysis used to screen patients and controls for the common haplogroup-associated polymorphisms showed that each of these changes was present in MS patients at a similar frequency to control subjects. However, 21 of the 58 patients (36.2%) were positive for T4 216C mutation, while only 11.3% of the controls carried this secondary LHON base change (P < 0.01; OR = 4.38). Our finding indicated that 4216C base substitution could be considered as a predisposing marker for MS and supported the hypothesis that particular mtDNA variants could contribute to genetic susceptibility of MS, and merits further investigation.

Association of actin with DNA and nuclear matrix from Guerin ascites tumour cells.

The protein composition of nuclear matrices containing different amount of DNA was examined. It was found that, in matrices containing 2% to 80% of total DNA, the quantity of DNA-bound proteins remains relatively constant varying from 10% to 15% of total nuclear proteins. Electrophoretic patterns do not differ substantially, but autoradiograms with in vitro 125I labelled proteins show quantitative variations in the actin content. Application of radioimmunoassay (RIA) enabled to determine the exact content of actin in GAT nuclei and nuclear matrices - 5 micrograms/ml in nuclei, of which 50% are bound to DNA and 30% being a component of the protein part of the nuclear matrix. These results are supported by electron microscopic data, where immunogold technique was performed on thin sections and spread material. The applied methods suggest that part of the nuclear actin is tightly bound (resistant to 2 M NaCl) to DNA and represents a component of the internal nuclear matrix.