Search and identification of thyroid hormones receptors in ocular tissues

Aim. To discover and identify thyroid receptors in ocular tissues by means of immunohistochemistry (IHC).Materials and methods.The objects of morphological studies were eyes enucleated for terminal painful glaucoma (n = 30). Thyroid receptors searching was performed in the retina and optic nerve, choroidal and optic nerve vessels. IHC reaction was considered as follows: negative — specific cellular staining is absent or less than 10 % cells are stained; mild positive — 10‑30 % cells are stained (+); moderate positive — 30‑75 % cells are stained (++); highly positive — more than 75 % are stained (+++). Results: Intensive (4+) IHC staining was revealed in the nuclei of inner and outer nuclear and ganglion cell layers. Mild positive (+) staining was detected in the inner segments of photoreceptors. Granular (3+) IHC staining was revealed in the nuclei of optic nerve glia. In choroidea, endotheliocytes nuclei and 20 % stromal cells nuclei were stained. IHCreaction was detected in the cytoplasm of retinal pigment epithelium.Conclusion. The data obtained account for the mechanism of neurohumoral associations at the cellular level. According to these data, eye can be considered as a target for thyroid hormones. The causes of morphological and functional abnormalities of visual analyzer peripheral part in thyroid gland disorders are revealed as well

Concerning the selection of areas with a dominant type of dependence when analyzing production control data

The formation of representative databases determines the interest in forecasting and managing the quality of metal based on data mining using special software products often based on regression analysis and not always taking into account the statistical nature of an object of study itself. This can lead to misinterpretation of the results or incomplete extracted information reducing the efficiency of statistical processing. Based on the analysis of the production database of the technology for producing 13G1S-U sheet steel, the authors evaluated the possibilities of multiple linear regression for predicting the quality of a steel sheet. The study shows that the type of distribution of the values of control parameters, the distribution nature of which was estimated based on the determination of the skewness and kurtosis coefficients, limits the regression forecast depth. Due to the great deviation of the predicted models from the experimental values in the right tail area of the distribution of the impact strength values, in this work, the authors developed the methods for separating data arrays and proposed criteria to compare the obtained results. To assess the accuracy of the results obtained, arrays with a deliberately asymmetric distribution were selected from the initial sample, against which the statistical characteristics were also compared. Based on the proposed techniques, the authors identified the dominant chemical elements that contribute to the difference in the distribution of the values of acceptance properties existing within the same standard technology. The study shows that the proposed separation method can be used as a variation of cognitive graphics techniques to identify areas with a dependence dominant type based on the correlation of skewness and kurtosis coefficients

Структурные особенности синтетических гликоконъюгатов и эффективность их взаимодействия с гликопротеиновыми рецепторами на поверхности гепатоцитов

Objectives. Over the last few years, medicinal chemistry research has been focusing on the creation of molecules that can target particular body systems, organs and tissues, thus abating systemic toxicity and side effects, and, most of all, boosting therapeutic potential. This goal can be achieved through the specific interaction of such drugs with active sites of cellular receptors. For example, glycoprotein receptors that can be found on cellular surfaces in neural tissues and liver parenchyma, selectively bind various glycoproteins and glycosides, facilitating their penetration into cells. This review describes how certain parameters of ligand structure (the nature and length of the spacer between carbohydrate and non-carbohydrate fragments of the molecule, number of carbohydrate residues per molecule, etc.) influence the penetration efficiency of synthetic glycoconjugates into liver cells.Methods. This review article summarizes 75 research papers and discusses data from in vitro and in vivo experiments showing which structures of synthetic carbohydrate derivatives are optimal for targeted drug delivery into liver cells.Results. The surface of liver cells (hepatocytes) contains a significant number of asialoglycoprotein receptors (ASGP-R) that are almost never found elsewhere. This makes ASGP-R an ideal target for the directed treatment of liver diseases, including such difficult, socially important conditions as hepatocellular carcinoma and Hepatitis C. A number of various ligands and targeted (to ASGP-R) delivery systems have been designed. Such molecules always contain derivatives of mono- and disaccharides, most commonly D-glucose, D-galactose, D-lactose and N-acetylglucosamines. This review contains the chemical structures of carbohydrate-based ligands.Conclusions. Glycolipids based on D-carbohydrates, when in liposomes, facilitate penetration into liver cells by a receptor-mediated, clathrin-dependent endocytosis mechanism that is activated upon contact of the carbohydrate-containing ligand fragment with the active site of ASGP-R. It can be addressed by the use of monovalent derivatives of carbohydrates as well as polyvalent glycoconjugates. Alterations in the ligand structure and the number of liposomal modifications can boost the therapeutic effect. The distance between the liposomal surface and the carbohydrate residue (spacer length), as well as the hydrophilic-lipophilic balance of the ligand molecule, have a great effect on the affinity and cellular response.Цели. Последние несколько лет исследования в области медицинской химии уделяют большое внимание созданию молекул, направленно воздействующих на конкретные системы организма человека, органы и ткани, что помогает снизить общее токсическое воздействие препаратов на их основе, уменьшить степень проявления побочных эффектов, а самое главное – многократно усилить их терапевтический эффект. Это может быть достигнуто при помощи специфического взаимодействия подобных веществ с активными центрами клеточных рецепторов. Например, класс гликопротеиновых рецепторов, располагающихся на поверхности клеток нервной ткани и паренхимы печени, селективно связывает различные гликопротеины и гликозиды, способствуя их проникновению внутрь клеток. В обзоре рассмотрено влияние таких особенностей структуры лигандов, как природа и длина связующего звена (спейсера) между углеводной и неуглеводной частями молекулы, количество углеводных остатков в составе одной молекулы, а также ряда других, на эффективность проникновения синтетических гликоконъюгатов в клетки печени.Методы. В обзоре проанализировано 75 публикаций и обобщены результаты исследований, в которых с помощью in vitro и in vivo экспериментов устанавливается, какая структура искусственно синтезированных производных углеводов окажется наиболее оптимальной для направленной доставки лекарственных средств в клетки печени.Результаты. На поверхности гепатоцитов (клеток печени) в большом количестве представлен асиалогликопротеиновый рецептор (ASGP-R), который почти не встречается на других типах клеток, что делает его идеальным рецептором-мишенью для направленного лечения заболеваний печени, в том числе таких трудно излечимых социально значимых заболеваний, как гепатоцеллюлярная карцинома и гепатит С. Разработан ряд разнообразных лигандов и систем направленной доставки к ASGP-R. Такие молекулы обязательно имеют в составе производные монои дисахаридов, чаще всего применяются D-глюкоза, D-галактоза, D-лактоза и N-ацетилглюкозамины. В обзоре приводятся примеры химических структур углеводсодержащих лигандов.Заключение. Гликолипиды на основе D-углеводов в составе липосом обеспечивают их проникновение в клетки печени по механизму рецептор-опосредованного клатрин-зависимого эндоцитоза, который активируется при контакте углеводсодержащей части лиганда с активным центром ASGP-R. Показано, что для этого можно использовать как моновалентные производные углеводов, так и поливалентные гликоконъюгаты. Варьируя структуру лиганда и количество добавляемых к липосоме модификаций, можно достичь наибольшего терапевтического эффекта. Большое влияние на аффинность и клеточный ответ оказывают расстояние от поверхности липосомы до углеводного остатка (длина спейсера) и гидрофильно-липофильный баланс молекулы лиганда

Синтез производных аминокислот на основе диэтаноламина с симметричными и асимметричными радикалами в гидрофобном домене с потенциальной антимикробной активностью

Objectives. Resistance to antibiotics and other antimicrobial drugs is an acute problem in the world today. Therefore, the chemical and pharmaceutical industries are still in search of new antibacterial agents that can overcome the resistance of pathogenic bacterial strains. To date, it has been established that molecules with antimicrobial activity must have an amphiphilic nature, a small size, one or more positive charges, and the required degree of hydrophobicity, that is, a significant hydrophilic–lipophilic balance (HLB) value. Some examples of such structures are antimicrobial peptides or peptidomimetics. This study aimed to develop a universal scheme for synthesizing several amino acid derivatives based on diethanolamine diesters with symmetric and asymmetric radicals in a hydrophobic block and potential antibacterial activity.Methods. The progression of chemical reactions was analyzed using thin-layer chromatography (TLC) on Sorbfil plates. The obtained compounds were isolated and purified using preparative TLC on Kieselgel (Merck) 60 F254 plates and column chromatography on Merck silica gel 0.040–0.063 mm. The TLC method was used to detect substances using a 3% ninhydrin solution, followed by heating to 70 °C. The structures of the obtained compounds were confirmed by hydrogen-1 nuclear magnetic resonance (1H NMR) spectroscopy on a Bruker WM-300 pulse NMR spectrometer, with hexamethyldisiloxane serving as the internal standard.Results. The HLB values of the diethanolamine derivatives were calculated, and samples were selected for subsequent synthesis. A scheme was developed for preparing amino acid derivatives based on diethanolamine diesters with symmetric and asymmetric radicals in the hydrophobic domain, and five new compounds were synthesized. The hydrophilic blocks of these compounds included residues of amino acids such as glycine, β-alanine, L-ornithine, and L-lysine.Conclusions. The potential antimicrobial activity of the synthesized peptidomimetics was assessed by their HLB values using the ACD/Labs Log P program. New amphiphiles were synthesized using amino acids and diethanolamine, and their structures were confirmed by 1H NMR spectroscopy data. The synthesized compounds were prepared for antibacterial activity analysis.Цели. Резистентность к антибиотикам и другим антимикробным препаратам является острой проблемой современности. Поиск новых антибактериальных агентов, преодолевающих резистентность патогенных штаммов бактерий, является востребованным в химико-фармацевтической промышленности. На сегодняшний день известно, что молекулы, обладающие антимикробной активностью, должны быть амфифильной природы, иметь небольшой размер, нести один или несколько положительных зарядов, а также обладать необходимой степенью гидрофобности, выраженной величиной гидрофильно-липофильного баланса. Такими структурами могут выступать антимикробные пептиды или пептидомиметики. Целью данного исследования была разработка универсальной схемы и проведение синтеза ряда производных аминокислот на основе диэфиров диэтаноламина с симметричными и асимметричными радикалами в гидрофобном блоке с потенциальной антибактериальной активностью.Методы. Анализ химических реакций, выделение и очистку полученных соединений проводили с помощью тонкослойной и колоночной хроматографии. Обнаружение веществ осуществляли методом тонкослойной хроматографии с использованием нингидриновой реакции для их визуализации на пластинах. Структуры полученных соединений подтверждали методом 1Н-ЯМР спектроскопии.Результаты. Проведен расчет величин гидрофильно-липофильного баланса производных диэтаноламина и выбраны образцы для последующего синтеза. Разработаны схемы получения производных аминокислот на основе диэфиров диэтаноламина с симметричными и асимметричными радикалами в гидрофобном домене и проведен синтез пяти новых соединений, в состав гидрофильных блоков которых входят остатки таких аминокислот, как глицин, β-аланин, L-орнитин и L-лизин.Выводы. Произведена оценка потенциальной антимикробной активности синтезированных пептидомиметиков по величине их гидрофильно-липофильного баланса с помощью программы ACD/Labs Log P. Синтезированы новые амфифилы на основе аминокислот и диэтаноламина, структуры которых подтверждены данными 1H-ЯМР спектроскопии

Literature review and clinical observation of acquired idiopathic hemophilia with a new missense mutation in the factor VIII gene (His2026Arg)

The article provides review of possible mechanisms of inhibitor coagulopathies, in particular of acquired hemophilia A. This pathology is an extremely rare disease occurring in 1-2 cases in 1 million per year. In the present study we provide data for two clinical cases of hemophilia A in women. These cases had different development mechanisms, although both women have a newly discovered missense mutation His2026Arg in the VIII factor gene. The matter of main interest is the description of the disease development in the patient with an acquired idiopathic hemophilia A with a possible disease occurrence due to an asymmetric X-chromosome inactivation (lyonization). In this particular case lyonization led to the late manifestation of the hemophilia A carrier’s state and development of severe form of the inhibitor-associated acquired hemophilia A. We also discuss therapeutic approaches to these forms of the disease, considering there are no concise protocols for case management due to an extreme rarity of the pathology. Acquainting the clinical personnel working it the different areas of medicine with suchlike inhibitor coagulopathies has a major practical importance

The association between <i>Candida albicans</i> sensitivity to antimycotic drugs and the architecture of their microbial community in the oropharynx of HIV infected patients

Relevance. Candida infection remains relevant due to the wide spread of antimycotic-resistant strains of Candida fungi, especially among immunocompromised individuals. It was previously discovered that the Candida spp. microbial community in the biotope of the oropharynx of HIV infected patients is characterized by a certain architecture: they can be present in this biotope in the form of a monoculture or as association of co-isolates. It has been suggested that the architecture of the Candida microbial community may influence their resistance to antimycotic drugs. Purpose a comparative study of the association between the architecture of the C. albicans microbial community in the oropharynx of HIV infected patients with oropharyngeal candidiasis and their sensitivity to antimycotic drugs. Materials and methods. A microbiological study of 52 isolates of Candida fungi (C. albicans, C. glabrata, C. tropicalis and C. krusei) from the oropharynx of 31 HIV infected patients with clinical manifestations of oropharyngeal candidiasis aged 20 to 69 years with almost equal gender distribution was carried out. In the form of monocultures, 18 isolates were isolated, while 34 were identified as co-isolates, which formed 16 homogeneous communities that included strains of the same species, and 18 heterogeneous ones that consisted of fungi of various species. Results. It was found that heterogeneous communities of C. albicans were markedly distinguished by sensitivity to antimycotic drugs, in particular, by low sensitivity to imidazoles. Homogeneous communities practically did not differ from monocultural ones. The general properties of the C. non-albicans population were largely similar to those of C. albicans, but were characterized by lower heterogeneity in response to antimycotic drugs. Conclusion. The architecture of the community of C. albicans isolated from the oropharynx of HIV infected patients with clinical manifestations of oropharyngeal candidiasis affects their sensitivity to antimycotic drugs. When selecting effective antimycotic therapy for such patients, it is necessary to take into account the structure of the Candida spp. community in the oropharynx

Морфологические основы глаз-тиреоидной ассоциации

PURPOSE: Search and identification of thyroid hormone receptors in the structures of the eye by means of immuno- histochemistry analysis (IHC). METHODS: Enucleated eyes of the patients with terminal painful glaucoma (n=30) were the object of morphological research. The search of thyroid hormone receptors was conducted in the retina and optic nerve, ciliary body and its appendixes, trabecular tissue, vessels of choroidea and optic nerve. Antibodies to thyroid hormone receptor THRalfa monoclonal («Diagnostic BioSystem» company, dilution 1:50) and rabbit polyclonal antibodies Anti-Thyroglobulin («Abcam» company, dilution 1:100) were used as primary antibodies (AT). IHC reaction was estimated as follows: - the absence of specific cell color or the presence of less than 10% of stained cells out of all examined area was considered a negative reaction; - a mild positive reaction was considered in case of 10 to 30% staining (+ cells); - moderate positive reaction was in case of the marker expression in 30-75% (++ cells); - marker expression that exceeded 75% was considered a pronounced reaction (+++); - intensive reaction - 100% stained cells (++++). RESULTS: Intensive (4+) positive IHC was discovered in the nuclei of external and internal granular layers of the retina, in the nuclei of the ganglion cells; mild positive staining (1+) was observed in the internal segments of the photoreceptor cells processes. Granule IHC staining (3+) was revealed in the nuclei of the optic nerve glial elements. In the choroidea endotheliocyte nuclei and nuclei of 20% of stroma cells of vascular membrane were stained. There was a marked IHC reaction in the cytoplasm of pigment epithelium cells. Positive staining was observed in the cytoplasm and the nuclei of trabecular endothelial cells, as well as in the nuclei of fibrocytes forming trabecular stroma. Nuclei staining of 80% smooth muscle cells with intensive 3+ and mild cytoplasmic staining 1+ was noted in the ciliary muscle. Pigment and amelanotic epithelium nuclei as well as the nuclei of vessel endothelium cells were stained intensively (3+) in the ciliary body proccesses. CONCLUSION: Conducted investigations help give a morphological base to eye-thyroid association, as well as explain the possible mechanisms of disthyroid optic neuropathy and ocular hypertension, and open-angle glaucoma prevalence in the population of patients with thyroid dysfunction.Цель. Поиск и идентификация рецепторов тиреоидных гормонов в структурах глаза методом иммуногистохимического анализа (ИГХ). методы. Объектом морфологических исследований стали энуклеированные глаза больных терминальной болящей глаукомой (n=30). Поиск рецепторов тиреоидных гормонов осуществляли в сетчатке и зрительном нерве, цилиарном теле и его отростках, в трабекулярной ткани, сосудах хориоидеи и зрительного нерва. Первичным антителом (АТ) служили АТ к рецепторам тиреоидных гормонов THRalfa monoclonal (фирма «Diagnostic BioSystem», разведение 1:50) и поликлональные кроличьи антитела Anti-Thyroglobulin (фирма «Abcam», разведение 1:100). ИГХ реакцию оценивали следующим образом: - отрицательной считали реакцию в отсутствие специфического окрашивания клеток или при наличии менее 10% окрашенных клеток от всей площади в зоне просмотра; - слабо положительной реакцию считали в случае окрашивания от 10 до 30% (+) клеток; - умеренно положительной считали реакцию в случае экспрессии маркера в 30-75% (++) клеток; - экспрессию маркера более 75% клеток расценивали как выраженную реакцию (+++); - интенсивная реакция - 100% окраска клеток (++++). РЕЗУЛЬТАТЫ. Интенсивная (4+) положительная ИГХ- окраска зафиксирована в ядрах наружного и внутреннего зернистых слоев сетчатки, в ядрах ганглиозных клеток; слабоположительная окраска (1+) - во внутренних сегментах отростков фоторецепторных клеток. Гранулярная ИГХ-окраска (3+) обнаруживалась в ядрах глиальных элементов зрительного нерва. В хориоидее окрашивались ядра эндотелиоцитов и ядра 20% клеток стромы сосудистой оболочки. ИГХ-реакция отмечалась в цитоплазме пигментного эпителия сетчатки. Положительная окраска зафиксирована в цитоплазме и ядрах эндотелиальных клеток трабекулярного аппарата, а также в ядрах фиброцитов, образующих строму трабекулы. В цилиарной мышце визуализировалась ядерная окраска 80% гладкомышечных клеток с интенсивностью 3+, а также слабая цитоплазматическая окраска 1+. В отростках цилиарного тела интенсивно окрашивались (3+) ядра пигментного и беспигментного эпителия, а также ядра эндотелиальных клеток сосудов. ЗАКЛЮЧЕНИЕ. Проведенные исследования позволяют подвести морфологические основы под глаз-тиреоидные ассоциации, объяснить возможные механизмы развития дистиреоидной оптической нейропатии и офтальмогипертензии, а также распространенность открытоугольной глаукомы в популяции больных с тиреоидной патологией

The structure of the oropharyngeal genus Candida fungi community in HIVinfected patients

At the present time virtually no data are available about the structure of the genus Candida fungus able to target HIV-infected patients and serve as an etiological factor of candidiasis. The aforementioned shaped the aim of the study: to examine structure of the Candida genus community colonizing the oropharynx in HIV-infected patients with clinical manifestations of oropharyngeal candidiasis. There was conducted a microbiological study of the oropharynx in 31 HIV-infected patients (51.6% males and 48.4% females) with clinical manifestations of oropharyngeal candidiasis treated at Moscow Infectious Clinic No. 2 inpatient department in the years 2015–2017. We confirmed the diversity of the oropharyngeal Candida spp. community found in HIV-infected patients. Total 52 isolates of the genus Candida were isolated. C. albicans dominated in 57.7% cases, whereas C. glabrata prevailed (21.1%) among non-albicans species. Minor components were represented by C. tropicalis (11.5%) and C. krusei (9.6%). C. albicans and C. glabrata were sensitive to polyenes, whereas minor community components — to itroconazole and clotrimazole. The vast majority of fungal strains were resistant to fluconazole. The genus Candida community reveals a unique architecture so that any member may exist in the oropharyngeal biotope of HIV-infected patients as a monoculture or in association: homogeneous, consisting of a single species strains, or heterogeneous, formed by several species. Candida fungi in 18 patients (58.1%) were isolated as a monoculture, whereas in 13 (41.9%) subjects — in association consisting of 34 isolates (65.4% of total number), of which 16 (30.8%) and 18 (34.6%) were isolated from homogeneous and heterogeneous associations, respectively. There were identified 9 two-component associations (69.2%), and 4 (30.8%) consisting of three or more components. It turned out that pattern of the examined community was mainly determined by species composition that agrees with previous data. Most common associations were presented by C. krusei (100%) and C. albicans (73.3%). Upon that, most often C. albicans (72.7%) formed a homogeneous type of associations. Sensitivity of Candida fungi to antimycotic drugs also depended on the architecture of related community. C. albicans isolates in heterogeneous associations revealed a wide range of resistance acquired by contact with non-albicans species

ЛЕГОЧНОЕ КРОВОТЕЧЕНИЕ: ЭНДОСКОПИЧЕСКАЯ ДИАГНОСТИКА И ЛЕЧЕНИЕ

In the article, we report the incidence and etiology of pulmonary hemorrhage, and modern classifications according to the literature data. Methods of endoscopic diagnosis and treatment of pulmonary hemorrhage are analyzed.В представленной статье на основе данных литературы приведены сведения о распространенности и этиологии легочных кровотечений (ЛК), изложены современные классификации. Проанализированы методы эндоскопической диагностики и лечения ЛК

Тетравалентный неогликоконъюгат на основе d-галактозы для целей медицины

Previously, the scheme of the preparation of a tetravalent neoglycoconjugate with the terminal residues of D-galactose and with a branching component based on D-galactose was carried out in our laboratory. It includes the synthesis of hydrophilic and hydrophobic components, the synthesis of the branching scaffold and its conjugation with a hydrophilic component. This article describes the optimization of the synthesis of 1-O-azidoethyl-β-D-galactopyronaside using the change of different parameters such as temperature, solvent and reaction time. The structure of obtained glycoconjugates has potential to be capable to anti-adhesion and target delivery to certain cell group.Проведена оптимизация ранее разработанной схемы синтеза тетравалентного неогликоконъюгата с терминальными остатками D-галактозы и разветвляющей компонентой на основе D-галактозы. Структура полученных гликоконъюгатов обеспечивает им потенциальную возможность проявлять свойства антиадгезии и специфически связываться с рецепторами на определенных группах клеток