The content of human sweat is studied by high-resolution NMR, and the majority of organic components most often found in sweat of conditionally healthy people are identified. Original and simple ...tools are designed for sweat sampling from different areas of human body. The minimal surface area needed for sampling is in the range of 50-100 cm(2). On all the surface parts of the human body examined in this work, the main constituents forming a sweat metabolic profile are lactate, glycerol, pyruvate, and serine. The only exception is the sole of the foot (planta pedis), where trace amounts of glycerol are found. An attempt is made to explain the presence of specified metabolites and their possible origin.
A giant multidomain protein of striated and smooth vertebrate muscles, titin, consists of tandems of immunoglobulin (Ig)- and fibronectin type III (FnIII)-like domains representing β-sandwiches, as ...well as of disordered segments. Chicken smooth muscles express several titin isoforms of ~500-1500 kDa. Using various structural-analysis methods, we investigated in vitro nonspecific amyloid aggregation of the high-molecular-weight isoform of chicken smooth-muscle titin (SMT
, ~1500 kDa). As confirmed by X-ray diffraction analysis, under near-physiological conditions, the protein formed amorphous amyloid aggregates with a quaternary cross-β structure within a relatively short time (~60 min). As shown by circular dichroism and Fourier-transform infrared spectroscopy, the quaternary cross-β structure-unlike other amyloidogenic proteins-formed without changes in the SMT
secondary structure. SMT
aggregates partially disaggregated upon increasing the ionic strength above the physiological level. Based on the data obtained, it is not the complete protein but its particular domains/segments that are likely involved in the formation of intermolecular interactions during SMT
amyloid aggregation. The discovered properties of titin position this protein as an object of interest for studying amyloid aggregation in vitro and expanding our views of the fundamentals of amyloidogenesis.
genus has been extensively studied due to its diverse biological activities. We evaluated the methanolic extract of leaves of
(Green type) (Thunb.) Jacques, the species that is less studied compared ...to
. The aim was to identify phytoconstituents of the methanolic extract of leaves of
and biological properties of its different fractions. Water fraction was analyzed with matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry. Nineteen compounds belonging to different chemical classes were identified in the methanolic extract of leaves of
(Green type) (Thunb.) Jacques. In addition to several fatty acids, isoprenoid and steroid compounds were found among the most abundant constituents. One of the identified compounds, 4'-methylphenyl-1C-sulfonyl-β-d-galactoside, was not detected earlier in
extracts. The water fraction was toxic to HeLa cells but not to Vero cells. Our data demonstrate that methanolic extract of leaves of
can be a valuable source of bioactive constituents. The water fraction of the extract exhibited promising antitumor potential based on a high ratio of HeLa vs. Vero cytotoxicity.
This work investigated in vitro aggregation and amyloid properties of skeletal myosin binding protein-C (sMyBP-C) interacting in vivo with proteins of thick and thin filaments in the sarcomeric ...A-disc. Dynamic light scattering (DLS) and transmission electron microscopy (TEM) found a rapid (5-10 min) formation of large (>2 μm) aggregates. sMyBP-C oligomers formed both at the initial 5-10 min and after 16 h of aggregation. Small angle X-ray scattering (SAXS) and DLS revealed sMyBP-C oligomers to consist of 7-10 monomers. TEM and atomic force microscopy (AFM) showed sMyBP-C to form amorphous aggregates (and, to a lesser degree, fibrillar structures) exhibiting no toxicity on cell culture. X-ray diffraction of sMyBP-C aggregates registered reflections attributed to a cross-β quaternary structure. Circular dichroism (CD) showed the formation of the amyloid-like structure to occur without changes in the sMyBP-C secondary structure. The obtained results indicating a high in vitro aggregability of sMyBP-C are, apparently, a consequence of structural features of the domain organization of proteins of this family. Formation of pathological amyloid or amyloid-like sMyBP-C aggregates in vivo is little probable due to amino-acid sequence low identity (<26%), alternating ordered/disordered regions in the protein molecule, and S-S bonds providing for general stability.
Cerebral ischemia, and, as a result, insult, attacks up to 15 million people yearly in the world. In this connection, the development of effective preventive programs and methods of therapy has ...become one of the most urgent problems in modern angiology and pharmacology. The cytoprotective action of taxifolin (TAX) in ischemia is well known, but its limitations are also known due to its poor solubility and low capacity to pass through the hematoencephalic barrier. Molecular mechanisms underlying the protective effect of TAX in complex systems such as the brain remain poorly understood. It is known that the main cell types of the brain are neurons, astrocytes, and microglia, which regulate the activity of each other through neuroglial interactions. In this work, a comparative study of cytoprotective mechanisms of the effect of TAX and its new water-soluble form aqua taxifolin (aqTAX) was performed on cultured brain cells under ischemia-like conditions (oxygen-glucose deprivation (OGD)) followed by the reoxygenation of the culture medium. The concentration dependences of the protective effects of both taxifolin forms were determined using fluorescence microscopy, PCR analysis, and vitality tests. It was found that TAX began to effectively inhibit necrosis and the late stages of apoptosis in the concentration range of 30-100 µg/mL, with aqTAX in the range of 10-30 µg/mL. At the level of gene expression, aqTAX affected a larger number of genes than TAX; enhanced the basic and OGD/R-induced expression of genes encoding ROS-scavenging proteins with a higher efficiency, as well as anti-inflammatory and antiapoptotic proteins; and lowered the level of excitatory glutamate receptors. As a result, aqTAX significantly inhibited the OGD-induced increase in the Ca
levels in the cytosol (Ca
) in neurons and astrocytes under ischemic conditions. After a 40 min preincubation of cells with aqTAX under hypoxic conditions, these Ca
signals were completely inhibited, resulting in an almost complete suppression of necrotic death of cerebral cortical cells, which was not observed with the use of classical TAX.
Among the vascular prostheses used for aortic replacement, 95% are woven or knitted grafts from polyester fibers. Such grafts require sealing, for which gelatin (Gel) is most often used. Sometimes ...antibiotics are added to the sealant. We used gelatin type A (GelA) or type B (GelB), containing one of the three antibiotics (Rifampicin, Ceftriaxone, or Vancomycin) in the sealant films. Our goal was to study the effect of these combinations on the mechanical and antibacterial properties and the cytocompatibility of the grafts. The mechanical characteristics were evaluated using water permeability and kinking radius. Antibacterial properties were studied using the disk diffusion method. Cytocompatibility with EA.hy926 endothelial cells was assessed via indirect cytotoxicity, cell adhesion, and viability upon direct contact with the samples (3, 7, and 14 days). Scanning electron microscopy (SEM) and energy dispersive spectrometry (EDS) were used to visualize the cells in the deep layers of the graft wall. "GelA + Vancomycin" and "GelB + vancomycin" grafts showed similar good mechanical characteristics (permeability~10 mL/min/cm
, kinking radius 21 mm) and antibacterial properties (inhibition zones for
~15 mm, for
~12 mm). The other samples did not exhibit any antibacterial properties. The cytocompatibility was good in all the tested groups, but endothelial cells exhibited the ability to self-organize capillary-like structures only when interacting with the "GelB + antibiotics" coatings. Based on the results obtained, we consider "GelB + vancomycin" sealant to be the most promising.
A comparative study of amyloid properties of the aggregates of smooth muscle titin (SMT) from chicken gizzard was carried out. These aggregates were formed in two solutions: 0.15 M glycine-KOH, pH ...7.2-7.4 (SMT(Gly)) and 0.2 M KCl, 10 mM imidazole, pH 7.0 (SMT(KCl)). Electron microscopy data showed that SMT aggregates has an amorphous structure in both cases. The results of atomic-force microscopy demonstrated slight differences in morphology in two types of aggregates. The SMT(Gly) aggregates were represented as branching chains, composed of spherical aggregates approximately 300-500 nm in diameter and up to 35 nm in height. The SMT(KCl) aggregates formed sponge-like structures with strands of 8-10 nm in height. Structural analysis of SMT aggregates by X-ray diffraction revealed the presence of cross-β-sheet structure in the samples under study. In the presence of SMT(Gly) aggregates, thioflavine T fluorescence intensity was higher (~3-fold times) compared with that in the presence of SMT(KCl) aggregates. Congo red-stained SMT(Gly) aggregates had yellow to apple-green birefringence under polarized light, which was not observed for SMT(KCl) aggregates. Dynamic light scattering data showed the similar rate of aggregation for both types of aggregates, though SMT(KCl) aggregates were able to partially disaggregate under increased ionic strength of the solution. The ability of SMT to aggregation followed by disaggregation may be functionally significant in the cell.
Two chimeric proteins, SHН and SHA of the “SH3-Bergerac” family (where the β-turn N47D48 in spectrin SH3 domain was substituted for
KITVNGKTYE or
KATANGKTYE sequences, respectively), were analyzed by ...high-resolution NMR to resolve their spatial structures and to analyze their dynamics. Although the presence of a stable β-hairpin in the region of the insertion was confirmed, the introduced extension of the polypeptide chain in SHН (∼
17%) practically did not affect the total molecule topology. Interestingly, the introduced β-hairpin had higher mobility in comparison with other protein regions. Finally, we performed a disorder prediction with the PONDR
® VSL2 algorithm and discovered that the inserted β-hairpin in both SHH and SHA proteins exhibited significant propensity for intrinsic disorder and therefore for high mobility. In agreement with the experimental data, the predisposition for the increased intramolecular mobility was noticeably higher in SHA.
As a result of research it was succeeded to receive the model of non-alcoholic steatohepatitis which is characterized by obesity, violation of the structural organization, a fabric necrosis, ...formation of fibrosis of a liver. The developed model of non-alcoholic steatohepatitis allows to study also staging of development of a fatty desease of the liver which is characterized by formation of a steatogepatitis and fibrosis. In quality the inducing factors the high-fat diet and antibiotics were used. The offered NASH model can be used in the field of chronic experiments for an assessment of extent of violation of a functional condition of hepatobiliary system and for search of the effective pharmacotheurapeutichepatoprotective agents.
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