Во многих тканях и клетках человека присутствуют как β1-, так и β2-адренорецепторы, информация о содержании и динамике поведения кото- рых часто является клинически значимой. В настоящем исследовании предложена методика раздельного определения обоих типов адреноре- цепторов на основе радиолигандного анализа с использованием 125I-йодоцианопиндолола, включающая проведение трех измерений: 1) без лигандов-конкурентов; 2) в присутствии селективного лиганда ICI 118,551 (0,25 мкМ); 3) в присутствии двух селективных лигандов ICI 118,551 и CGP 20712 (по 0,25 мкМ каждого). Методика протестирована на модельной системе из двух трансгенных линий клеток с экспрессией реком- бинантных β1- и β2-адренорецепторов. При соотношении количества β1-адренорецепторов к β2-адренорецепторам 1:10 погрешность измере- ния составляет около 15%. Анализ 9 клеточных линий, представляющих различные типы клеток крови, показал наличие β2-адренорецепторов в клетках Daudi, Raji, Dami, K-562, HL-60, U-937 и THP-1 и их отсутствие в Т-лимфоцитарных клетках Jurkat и MOLT-4. β1-адренорецепторы достоверно зарегистрированы лишь в клетках THP-1 моноцитарного происхождения. В остальных клетках, за исключением линии Dami, их количество оказалось ниже порога детекции, оцениваемого на уровне 250 молекул на клетку. Измерения, выполненные на мононуклеарных клетках периферической крови здоровых доноров, продемонстрировали присутствие β2-адренорецепторов в диапазоне от 1000 до 2500 моле- кул на клетку, тогда как содержание β1-адренорецепторов во всех случаях находилось на грани или за гранью порога детекции. По-видимому, изучение β1-адренорецепторов в дальнейшем следует проводить на отдельных фракциях клеток крови, в частности на фракции моноцитов β1- and β2-adrenergic receptors are presented in various human tissues and cells, while the information of their content and dynamic behavior is oftenly considered as clinically significant. In this study, a method for the separate determination of both types of adrenoceptors based on radioligand binding analysis using 125I iodocyanopindolol is proposed, comprising three measurements: 1) without competing ligands; 2) in the presence of selective ligand ICI 118,551 (0.25 μM); 3) in the presence of two selective ligands ICI 118,551 and CGP 20712 (0.25 μM each). The technique was tested on a model system of two transgenic cell lines with the expression of recombinant β1- and β2-adrenergic receptors. If the ratio of the number of β1-adrenergic receptors to β2-adrenergic receptors is 1:10, the measurement error is about 15%. Analysis of 9 cell lines representing different types of blood cells showed the presence of β2-adrenergic receptors in Daudi, Raji, Dami, K-562, HL-60, U-937 and THP-1 cells and their absence in Jurkat and MOLT-4 cells. β1-adrenergic receptors are reliably registered only in THP-1 cells of monocytic origin. In the remaining cell lines, with the exception of Dami, the number of β1-adrenergic receptors was found below the detection limit, estimated as 250 molecules per cell. Measurements performed on the peripheral blood mononuclear cells of healthy donors showed the presence of β2-adrenergic receptors in the range from 1000 to 2500 molecules per cell, while the content of β1-adrenergic receptors in all cases appeared to be on the border or beyond the detection limit. Apparently, further study of β1-adrenergic receptors should be performed on the blood cells isolated fractions, on monocytes in particular
T-cadherin is a unique member of the cadherin superfamily of adhesion molecules. In contrast to "classical" cadherins, T-cadherin lacks transmembrane and cytoplasmic domains and is anchored to the cell membrane via a glycosilphosphoinositol moiety. T-cadherin is predominantly expressed in cardiovascular system. Clinical and biochemical studies evidence that expression of T-cadherin increases in post-angioplasty restenosis and atherosclerotic lesions—conditions associated with endothelial dysfunction and pathological expression of adhesion molecules. Here, we provide data suggesting a new signaling mechanism by which T-cadherin regulates endothelial permeability. T-cadherin overexpression leads to VE-cadherin phosphorylation on Y731 (β-catenin-binding site), VE-cadherin clathrin-dependent endocytosis and its degradation in lysosomes. Moreover, T-cadherin overexpression results in activation of Rho GTPases signaling and actin stress fiber formation. Thus, T-cadherin up-regulation is involved in degradation of a key endothelial adhesion molecule, VE-cadherin, resulting in the disruption of endothelial barrier function. Our results point to the role of T-cadherin in regulation of endothelial permeability and its possible engagement in endothelial dysfunction.
The objective: to develop a new method to detect early subclinical signs of LTBI progression to the active disease. Subjects and Methods. Children under 18 years of age were enrolled in the study, of them 8 children suffered from pulmonary tuberculosis (TB Group) and 91 had LTBI (LTBI Group). Detection of subclinical signs of tuberculosis infection activity in LTBI Group included testing the concentration of antigen-specific induction of IFN-γ at the level more than 14 pg/ml, IL-6 more than 8 pg/ml and positive values of threshold cycles (Ct) of mRNA samples based on the results of real-time PCR of genes: PDCD1, PDL2, BATF2 and GBP5. Results. The developed method for detection of subclinical signs of tuberculosis infection activity in children and adolescents with LTBI consists of two components (immunological testing of concentration of IFN-γ and IL-6; and molecular genetic testing of mRNA expression of the PDL2, BATF2 and GBP5 genes). Simultaneous registration of IFN-γ and IL-6 levels and mRNA expression of the PDL2, BATF2 and GBP5 genes above reference values indicates a high risk of developing active tuberculosis. It was observed in 29/91 (31.9%) patients with LTBI, in whom chest CT revealed calcifications in the upper lymph nodes or pulmonary lesions within 1 year.
Water-soluble cationic polypyridylphenylene dendrimers (wPPPDs) of different generations and various contents of pyridyl and phenylene moieties were synthesized. Interaction of wPPPDs with DNA resulted in the formation of polyelectrolyte complexes stable at physiological pH and ionic strength. Noticeable contribution of hydrophobic interactions provided by phenylene groups of the dendrimer in the complex stability was ascertained. Data obtained by turbidimetry and a sedimentation assay, as well as ζ-potential measurements, strongly implied the inaccessibility of positively charged pyridinium groups situated in the inner part of the dendrimer molecules for interaction with the rigid double helix. Positively and negatively charged water-soluble nonstoichiometric polyelectrolyte complexes were prepared and phase separations in their water-salt solutions were investigated. According to DLS measurements supported by the sedimentation assay, the addition of wPPPD in DNA solution in a wide range of the mixture composition (before phase separation) led to two coexisting populations of nanoparticles related to the soluble DNA/wPPPDs complex and practically unbound DNA. The revealed spontaneous formation of nanoparticles of the positively charged soluble DNA/wPPPDs complexes stable at ionic strength close to the physiological one is extremely encouraging for the development of the wPPPD vehicle to deliver gene materials to the cell.
Abstract β-adrenoreceptor (ADRB) ligands are actively used in the therapy of bronchopulmonary and cardiovascular diseases. When using these drugs, it is important to assess changes in ADRB content in different tissues, ADRB1 in cardiovascular disease and ADRB2 in bronchopulmonary disease. Direct measurement of ADRB content in lung and heart cells is not possible in most cases. It was previously shown that ADRB2 content in peripheral blood lymphocytes (or mononuclear cells) is significantly correlated with that in myocardial cells. It has been suggested that blood lymphocytes can be used to monitor ADRB content in solid organs. However, estimation of ADRB1 content in myocardium from the content of this receptor in peripheral lymphocytes is not possible because of the low content of ADRB1 in lymphocytes. In the present study we performed simultaneous determination of ADRB1 and ADRB2 both in the total population of peripheral blood mononuclear cells (PBMCs) and in isolated subpopulations of monocytes, T-lymphocytes and NK-cells from 23 healthy donors using the radioligand method modified by us earlier. The highest amount of ADRB2 was detected in NK cells, followed by PBMCs, monocytes and T cells (in descending order). The content of these receptors on all blood cell subpopulations was significantly correlated with each other, suggesting the possibility of using PBMCs to monitor ADRB2 in solid organs. We first detected the presence of ADRB1 in monocytes in 43% of donors and in NK cells in 35% of donors.
'/%3e%3cpath id='b' d='M658.84 441.31c-7.618 16.446-22.845 19.271-36.164 5.995 0 0 5.354-3.783 11.086-4.826-1.075-4.574 1.13-10.902 4.235-14.324 3.258 2.227 7.804 6.689 8.96 11.874 8.03-1.04 11.883 1.282 11.883 1.282z'/%3e%3cuse xlink:href='%23b' transform='rotate(9.37 700 804)'/%3e%3cuse xlink:href='%23b' transform='rotate(18.74 700 804)'/%3e%3cpath fill='none' stroke='%23f8c300' stroke-width='16' d='M603 478a340 340 0 0 1 194 0'/%3e%3cg transform='translate(700 380)'%3e%3cg transform='translate(0 -140)'%3e%3cpath id='c' d='m0-513674 301930 929245-790463-574305h977066l-790463 574305z' transform='scale(.00005)'/%3e%3c/g%3e%3cg id='d'%3e%3cuse xlink:href='%23c' transform='translate(-70 -121.244)'/%3e%3cuse xlink:href='%23c' transform='translate(-121.244 -70)'/%3e%3cuse xlink:href='%23c' transform='translate(-140)'/%3e%3c/g%3e%3cuse xlink:href='%23d' transform='scale(-1 1)'/%3e%3c/g%3e%3c/g%3e%3c/svg%3e)
'/%3e%3c/g%3e%3cuse xlink:href='%23b' transform='matrix(1 0 0 -1 0 549)'/%3e%3c/svg%3e)