Human Developmental Chondrogenesis as a Basis for Engineering Chondrocytes from Pluripotent Stem Cells

Joint injury and osteoarthritis affect millions of people worldwide, but attempts to generate articular cartilage using adult stem/progenitor cells have been unsuccessful. We hypothesized that recapitulation of the human developmental chondrogenic program using pluripotent stem cells (PSCs) may represent a superior approach for cartilage restoration. Using laser-capture microdissection followed by microarray analysis, we first defined a surface phenotype (CD166(low/neg)CD146(low/neg)CD73(+)CD44(low)BMPR1B(+)) distinguishing the earliest cartilage committed cells (prechondrocytes) at 5-6 weeks of development. Functional studies confirmed these cells are chondrocyte progenitors. From 12 weeks, only the superficial layers of articular cartilage were enriched in cells with this progenitor phenotype. Isolation of cells with a similar immunophenotype from differentiating human PSCs revealed a population of CD166(low/neg)BMPR1B(+) putative cartilage-committed progenitors. Taken as a whole, these data define a developmental approach for the generation of highly purified functional human chondrocytes from PSCs that could enable substantial progress in cartilage tissue engineering.Fil: Wu, Ling. University of California at Los Angeles; Estados UnidosFil: Bluguermann, Carolina. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia. Laboratorio de Biología del Desarrollo Celular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. University of California at Los Angeles; Estados UnidosFil: Kyupelyan, Levon. University of California at Los Angeles; Estados UnidosFil: Latour, Brooke. University of California at Los Angeles; Estados UnidosFil: Gonzalez, Stephanie. University of California at Los Angeles; Estados UnidosFil: Shah, Saumya. University of California at Los Angeles; Estados UnidosFil: Galic, Zoran. University of California at Los Angeles; Estados UnidosFil: Ge, Sundi. University of California at Los Angeles; Estados UnidosFil: Zhu, Yuhua. University of California at Los Angeles; Estados UnidosFil: Petrigliano, Frank A.. University of California at Los Angeles; Estados UnidosFil: Nsair, Ali. University of California at Los Angeles; Estados UnidosFil: Miriuka, Santiago Gabriel. Fundación para la Lucha contra las Enfermedades Neurológicas de la Infancia. Laboratorio de Biología del Desarrollo Celular; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Li, Xinmin. University of California at Los Angeles; Estados UnidosFil: Lyons, Karen M.. University of California at Los Angeles; Estados UnidosFil: Crooks, Gay M.. University of California at Los Angeles; Estados UnidosFil: McAllister, David R.. University of California at Los Angeles; Estados UnidosFil: Van Handel, Ben. Novogenix Laboratories; Estados UnidosFil: Adams, John S.. University of California at Los Angeles; Estados UnidosFil: Evseenko, Denis. University of California at Los Angeles; Estados Unido

Evaluation of pharmacological efficiency of Omacor in patients with coronary heart disease with hyperlipidemia in combination with rhythm disorders

The article discusses the treatment of patients with coronary heart disease with hyperlipidemia and extrasystole omega-3 polyunsaturated fatty acids (omacor), which have both antiarrhythmic effects and normalize lipid metabolism, preventing the development of atherosclerosis. As a result of the study, positive changes were detected in the lipoprotein spectrum of blood plasma during pharmacotherapy with omacor, in particular the hypotriglyceridemic effect in combination with a significant increase in the level of high-density lipoproteins. There was also a decrease in the incidence of episodes of both ventricular and supraventricular extrasystole, which made it possible to use omacor in patients with coronary heart disease with post-infarction cardiosclerosis in combination with clinically significant extrasystole. The significant hypolidemic effect of omacor, as well as its antiarrhythmic effect on the severity of ventricular and supraventricular extrasystoles make its use for the correction of type IIB and type IV hyperlipidemia in combination with arrhythmias the most justified.В статье рассмотрены вопросы лечения больных ИБС с гиперлипидемией и экстрасистолией омега-3 полиненасыщенными жирными кислотами (омакор), обладающими как антиаритмическим действием, так и нормализующими липидный обмен, осуществляя профилактику развития атеросклероза. В результате проведённого исследования были выявлены положительные изменения в липопротеиновом спектре плазмы крови при фармакотерапии омакором, в частности гипотриглицеридемический эффект в сочетании с достоверным повышением уровня липопротеидов высокой плотности. Также было отмечено снижение частоты возникновения эпизодов как желудочковой, так и наджелудочковой экстрасистолии, что обусловило возможность применения омакора у больных ишемической болезнью сердца с постинфарктным кардиосклерозом в сочетании с клинически значимой экстрасистолией. Значимые гиполидемический и антиаритмический эффекты омакора делают его применение для коррекции IIБ и IV типа гиперлипидемии в сочетании с аритмиями наиболее оправданным

The efficacy of complex solid dispersions of anthelmintics against experimental trichinellosis

The purpose of the research is to study the influence of various technological factors on obtaining of complex solid dispersions of anthelmintics with polyvinylpyrrolidone and licorice extract on anthelmintic efficacy in experimental trichinellosis of white mice.Materials and methods. The study of the nematodocidal activity of complex solid dispersions samples based on fenbendazole (FBZ), fenasal (FNS) and praziquantel (PZQ) with polyvinylpyrrolidone (PVP) and licorice extract (LE) obtained by mechanochemical technology at different ratios of components and different exposure times was carried out on 130 white mice experimentally infected with Trichinella spiralis in two experiments. On the 3rd day after infection, the animals were divided into experimental groups of 10 animals each. Samples of various complex solid dispersions of anthelmintics were administered intragastrically to the mice of the experimental groups at a dose of 2 mg/kg according to the active substance. FBZ substance was used as the basic drug at a dose of 2 mg/kg according to the active substance. Animals of the control groups did not receive the drugs. The animals were killed by decapitation on the 4th day after experimental drug samples administration, and the activity of the drugs was counted according to the results of helminthological necropsy of the intestine, the efficacy was calculated by the type of control test.Results and discussion. The efficacy of complex solid dispersions of FBZ and FNS with PVP polymer was higher in comparison with the activity of complexes with LE at the same duration of mechanochemical treatment in a roller mill. The FBZ activity decreased from 67.05 to 37.77% with a decrease in the duration of mechanochemical treatment from 24 h to 5 h and the efficacy of the FBZ : FNS complex with LE turned out to be almost at the level of the basic drug when treated for 1 h. The use of mechanochemical technology for obtaining of a solid dispersion of FBZ : FNS with PVP for targeted delivery makes it possible to increase the anthelmintic efficacy by 2.7 times compared with the activity of the FBZ substance, and with LE by 2.2 times. It was noted that complex solid dispersions of PBZ with PZQ have lower biological activity in comparison with compositions of FBZ with FNS

Development and laboratory production of virus-like immune-stimulating complexes based on saponins and evaluation of their adjuvant potential using mice immunisation with influenza antigens

The COVID-19 pandemic has exacerbated the public’s need for effective vaccines. Consequently, significant financial support has been provided to developers of a number of innovative vaccines, including the vaccines with saponin-based adjuvants. In 2021, the World Health Organisation recommended Mosquirix, the first malaria vaccine, which contains a saponin adjuvant. An anti-covid vaccine by Novavax is in the approval phase. A promising approach to vaccine development is presented by the use of virus-like immune-stimulating complexes (ISCOMs) containing saponins and by the creation of combinations of ISCOMs with antigens. The aim of the study was to develop, produce and characterise virus-like immune-stimulating complexes based on saponins of Quillaja saponaria, as well as similar saponins of Russian-sourced Polemonium caeruleum. Materials and methods: The ISCOM adjuvants, Matrix-BQ and Matrix-BP, were produced using liquid chromatography and examined using electron microscopy. Balb/c mice were immunised intraperitoneally and intramuscularly with ISCOM-antigen preparations. Afterwards, the immunised animals were challenged with the influenza virus strain, A/California/4/2009(H1N1)pdm09, adapted and lethal to mice. The serum samples were examined using haemagglutination inhibition (HI) tests. Results: The authors produced the ISCOMs containing saponins of Quillaja saponaria and Polemonium caeruleum. After one intramuscular injection of either of the ISCOM-antigen preparations with 1 µg of each of A/Brisbane/02/2018 (H1N1) pdm09, A/Kansas/14/2017 (H3N2), and B/Phuket/3073/2013 haemagglutinin antigens (HAs), HI tests detected serum antibody titres to the corresponding antigens of ≥1:40. Two intramuscular injections of the ISCOM-antigen preparation containing 50 ng of each of the HAs and Matrix-BQ resulted in a protective response. In some animals, two intraperitoneal injections of ISCOM-antigen preparations resulted in the maximum antibody titre to the A/Kansas/14/2017 (H3N2) vaccine strain of 1:20,480. Two intramuscular injections of a test preparation containing 5 µg, 1 µg, 200 ng, or 50 ng of each of the HAs and Matrix-BQ or a control preparation containing 5 µg, 1 µg, or 200 ng of each of the HAs (commercially available vaccines) to the mice that were afterwards infected with the lethal influenza strain protected the experimental animals from death. Conclusions: The ISCOM-based preparations had high immunostimulatory activity in the mouse-model study. The presented results indicate the potential of further studies of ISCOM-based preparations in terms of both vaccine and immunotherapeutic development

Development of the Structure of it Support for the Process of Designing a Fuel Consumption Control System in Liquid Rocket Engines

Development of an IT support structure for the process of designing a fuel consumption management system in liquid rocket engines, which is a mathematical model of the SURT and a service where the operation of the system, the process of processing and storing data, displaying test results on graphs

Reduction of hepatotoxicity of nimesulide in mechanochemically obtained composition with disodium salt of glycyrrhizic acid

Nimesulide (NIM) is a nonsteroid anti-inflammatory drug which acts as a selective cyclooxygenase 2 inhibitor and is widely used for acute pain treatment. In medical practice, a large amount of data has been collected describing the effect of NIM on the body, while a hepatotoxic side effect of the drug has been found. The exact mechanisms of such NIM-induced hepatotoxicity largely remain unknown but likely involve the intermediate reaction of its metabolism. Reduction of the hepatotoxic side effect of NIM is an actual problem for pharmacology. The aim of the present research was to evaluate the hepatotoxicity of the mechanochemically obtained composition of NIM with glycyrrhizic acid disodium salt (Na2GA) compared to pure NIM and a physical mixture of NIM with Na2GA. Material and methods. CD-1 mice were orally administered for 14 days: 1 group – mechanochemical composition NIM/Na2GA (1:10, m/m) at a dose of 1650 mg/kg; 2 group – physical mixture of NIM with Na2GA (1:10, m/m) at a dose of 1650 mg/kg; 3 group – pure NIM at a dose of 600 mg/kg (which pharmacokinetically corresponds to 1650 mg/kg of NIM/Na2GA); 4 group – vehicle (distilled water). The liver damage was assessed using histological studies and enzymatic activity of the alanine aminotransferase and aspartate aminotransferase in blood serum. Results. Histological analysis did not detect any changes in the liver of NIM/Na2GA-treated animals in comparison with a water-treated group. On the opposite, NIM given alone or as a physical mixture with Na2GA induced severe hepatotoxicity in experimental mice. Biochemical analysis of the blood serum revealed that mechanochemical NIM/Na2GA composition significantly reduced activity of the alanine aminotransferase (about 1.5 times) and aspartate aminotransferase (1.3 times) as compared with the pure NIM. Conclusions. The results obtained indicate a high potential for the practical application of the NIM/Na2GA mechanochemical composition

Эффективность комплексных твердых дисперсий антигельминтиков при экспериментальном трихинеллезе

The purpose of the research is to study the influence of various technological factors on obtaining of complex solid dispersions of anthelmintics with polyvinylpyrrolidone and licorice extract on anthelmintic efficacy in experimental trichinellosis of white mice.Materials and methods. The study of the nematodocidal activity of complex solid dispersions samples based on fenbendazole (FBZ), fenasal (FNS) and praziquantel (PZQ) with polyvinylpyrrolidone (PVP) and licorice extract (LE) obtained by mechanochemical technology at different ratios of components and different exposure times was carried out on 130 white mice experimentally infected with Trichinella spiralis in two experiments. On the 3rd day after infection, the animals were divided into experimental groups of 10 animals each. Samples of various complex solid dispersions of anthelmintics were administered intragastrically to the mice of the experimental groups at a dose of 2 mg/kg according to the active substance. FBZ substance was used as the basic drug at a dose of 2 mg/kg according to the active substance. Animals of the control groups did not receive the drugs. The animals were killed by decapitation on the 4th day after experimental drug samples administration, and the activity of the drugs was counted according to the results of helminthological necropsy of the intestine, the efficacy was calculated by the type of control test.Results and discussion. The efficacy of complex solid dispersions of FBZ and FNS with PVP polymer was higher in comparison with the activity of complexes with LE at the same duration of mechanochemical treatment in a roller mill. The FBZ activity decreased from 67.05 to 37.77% with a decrease in the duration of mechanochemical treatment from 24 h to 5 h and the efficacy of the FBZ : FNS complex with LE turned out to be almost at the level of the basic drug when treated for 1 h. The use of mechanochemical technology for obtaining of a solid dispersion of FBZ : FNS with PVP for targeted delivery makes it possible to increase the anthelmintic efficacy by 2.7 times compared with the activity of the FBZ substance, and with LE by 2.2 times. It was noted that complex solid dispersions of PBZ with PZQ have lower biological activity in comparison with compositions of FBZ with FNS.Цель исследований – изучить влияние различных факторов технологии получения комплексных твердых дисперсий антигельминтиков с поливинилпирролидоном и экстрактом солодки на эффективность при экспериментальном трихинеллезе белых мышей.Материалы и методы. Изучение нематодоцидной активности образцов комплексных твердых дисперсий на основе фенбендазола (ФБЗ), фенасала (ФНС) и празиквантела (ПЗК) с поливинилпирролидоном (ПВП) и экстрактом солодки (ЭС), полученных по механохимической технологии при разном соотношении компонентов и различной продолжительности механообработки проводили на 130 белых мышах, экспериментально зараженных Trichinella spiralis в двух опытах. На третьи сутки после заражения животных разделили на группы по 10 голов в каждой. Мышам опытных групп вводили в желудок образцы различных комплексных твердых дисперсий антигельминтиков в дозе 2 мг/кг по ДВ. В качестве базового препарата использовали субстанцию ФБЗ в дозе 2 мг/кг по ДВ. Животные контрольных групп препараты не получали. На четвертые сутки после введения опытных образцов животных убивали декапитацией и активность препаратов учитывали по результатам гельминтологического вскрытия кишечника; эффективность рассчитывали по типу «контрольный тест».Результаты и обсуждение. Эффективность комплексных твердых дисперсий ФБЗ и ФНС с полимером ПВП была выше по сравнению с активностью комплексов с ЭС при одинаковой продолжительности механохимической обработки в валковой мельнице. С уменьшением продолжительности механохимической обработки с 24 ч до 5 ч активность ФБЗ снижалась с 67,05 до 37,77%, а при обработке в течение 1 ч эффективность комплекса ФБЗ : ФНС с ЭС оказалась практически на уровне базового препарата. Использование механохимической технологии получения твердой дисперсии ФБЗ : ФНС с ПВП позволяет повысить антигельминтную эффективность в 2,7 раза по сравнению с активностью субстанции ФБЗ, а с ЭС – в 2,2 раза. Отмечено, что комплексные твердые дисперсии ФБЗ с ПЗК обладают меньшей биологической активностью в сравнении с композициями ФБЗ с ФНС

Разработка и лабораторное получение вирусоподобных иммуностимулирующих комплексов на основе сапонинов, оценка их адъювантных свойств при иммунизации мышей гриппозными антигенами

The COVID-19 pandemic has exacerbated the public’s need for effective vaccines. Consequently, significant financial support has been provided to developers of a number of innovative vaccines, including the vaccines with saponin-based adjuvants. In 2021, the World Health Organisation recommended Mosquirix, the first malaria vaccine, which contains a saponin adjuvant. An anti-covid vaccine by Novavax is in the approval phase. A promising approach to vaccine development is presented by the use of virus-like immune-stimulating complexes (ISCOMs) containing saponins and by the creation of combinations of ISCOMs with antigens. The aim of the study was to develop, produce and characterise virus-like immune-stimulating complexes based on saponins of Quillaja saponaria, as well as similar saponins of Russian-sourced Polemonium caeruleum. Materials and methods: The ISCOM adjuvants, Matrix-BQ and Matrix-BP, were produced using liquid chromatography and examined using electron microscopy. Balb/c mice were immunised intraperitoneally and intramuscularly with ISCOM-antigen preparations. Afterwards, the immunised animals were challenged with the influenza virus strain, A/California/4/2009(H1N1)pdm09, adapted and lethal to mice. The serum samples were examined using haemagglutination inhibition (HI) tests. Results: The authors produced the ISCOMs containing saponins of Quillaja saponaria and Polemonium caeruleum. After one intramuscular injection of either of the ISCOM-antigen preparations with 1 µg of each of A/Brisbane/02/2018 (H1N1) pdm09, A/Kansas/14/2017 (H3N2), and B/Phuket/3073/2013 haemagglutinin antigens (HAs), HI tests detected serum antibody titres to the corresponding antigens of ≥1:40. Two intramuscular injections of the ISCOM-antigen preparation containing 50 ng of each of the HAs and Matrix-BQ resulted in a protective response. In some animals, two intraperitoneal injections of ISCOM-antigen preparations resulted in the maximum antibody titre to the A/Kansas/14/2017 (H3N2) vaccine strain of 1:20,480. Two intramuscular injections of a test preparation containing 5 µg, 1 µg, 200 ng, or 50 ng of each of the HAs and Matrix-BQ or a control preparation containing 5 µg, 1 µg, or 200 ng of each of the HAs (commercially available vaccines) to the mice that were afterwards infected with the lethal influenza strain protected the experimental animals from death. Conclusions: The ISCOM-based preparations had high immunostimulatory activity in the mouse-model study. The presented results indicate the potential of further studies of ISCOM-based preparations in terms of both vaccine and immunotherapeutic development.Пандемия COVID-19 обострила потребность общества в эффективных вакцинных препаратах. В этих условиях существенную финансовую поддержку получили разработчики ряда инновационных вакцин, в том числе вакцин, в состав которых входят адъюванты на основе сапонинов. В 2021 г. ВОЗ была одобрена первая противомалярийная вакцина Mosquirix, содержащая сапонины. На стадии одобрения находится вакцина Novavax против COVID-19. Перспективным подходом к созданию вакцин является использование вирусоподобных иммуностимулирующих комплексов (ИСКОМ) на основе сапонинов и создание на их основе комплексов с антигеном (ИСКОМ-антиген). Цель работы: получение и изучение вирусоподобных иммуностимулирующих комплексов на основе сапонинов Квиллайи мыльной (Quillaja saponaria), а также аналогов на основе сапонинов Синюхи голубой (Polemonium caeruleum), полученных из отечественного сырья. Материалы и методы: с применением метода жидкостной хроматографии получали препараты ИСКОМ адъювантов — Матрикс-BQ и Матрикс-BP. Проведено электронно-микроскопическое исследование препаратов. Иммунизацию мышей Balb/c препаратами ИСКОМ-антиген проводили интраперитонеально и внутримышечно. Иммунизированных животных заражали адаптированным летальным для мышей штаммом вируса гриппа A/California/4/2009 (H1N1) pdm09. Образцы сыворотки крови иммунизированных животных исследовали в реакции торможения гемагглютинации (РТГА). Результаты: получены ИСКОМ, содержащие сапонины Синюхи голубой и Квиллайи мыльной. В образцах сыворотки крови животных, однократно внутримышечно иммунизированных препаратом ИСКОМ-антиген, содержащим по 1 мкг гемагглютинина каждого из штаммов вирусов гриппа A/Brisbane/02/2018 (H1N1) pdm09, A/Kansas/14/2017 (H3N2), B/ Phuket/3073/2013, значения титров антител в РТГА составили более 1:40 к соответствующим антигенам. При двукратном внутримышечном введении препарата ИСКОМ-антиген, содержащего 50 нг каждого антигена, был выявлен протективный ответ. Максимальные значения титров антител в РТГА выявлены при двукратном интраперитонеальном введении препарата ИСКОМ-антиген и составили 1:20480 к гемагглютинину вакцинного штамма A/Kansas/14/2017 (H3N2). Показано, что двукратное внутримышечное введение 5 мкг, 1 мкг, 200 нг, 50 нг препарата ИСКОМ-антиген и 5 мкг, 1 мкг, 200 нг контрольного антигена коммерчески доступной вакцины мышам, впоследствии зараженным летальным штаммом вируса гриппа A/California/4/2009 (H1N1)pdm09, защищает экспериментальных животных от гибели. Выводы: полученные препараты на основе ИСКОМ обладали высокой иммуностимулирующей активностью в исследовании на мышиной модели. Представленные результаты свидетельствуют о перспективности дальнейшего изучения препаратов на основе ИСКОМ при разработке как противовирусных, так и иммунокорректирующих препаратов

Influenza (H5N1) Viruses in Poultry, Russian Federation, 2005–2006

Migrating waterfowl may be the primary source of influenza (H5N1) in western Siberia and the European part of the Russian Federation

CHARACTERIZATION OF AVIAN INFLUENZA H5N8 VIRUS STRAINS THAT CAUSED THE OUTBREAKS IN THE RUSSIAN FEDERATION IN 2016–2017

Objective of the study is to investigate biological properties of avian influenza virus strains that caused the outbreaks in Russia in 2016–2017.Materials and methods. The study was performed using advanced virological and molecular-biological methods in state-of-the-art equipment.Results and conclusion. In 2016, the outbreaks among wild birds and poultry caused by highly pathogenic avian influenza H5N8 virus have occurred in the territory of the Russian Federation. In May, 2016 an outbreak of H5N8 among wild birds was registered in the territory of the Republic of Tyva. In October-November, 2016 influenza virus H5N8 was isolated in the territory of the Republics of Tatarstan and Kalmykia, Krasnodar and Astrakhan Regions of Russia. In 2017 avian influenza H5N8 has become widespread in European part of Russia and caused multiple outbreaks among wild birds and poultry. Results of the investigations of the isolated strains show that all of them are highly pathogenic and belong to the clade 2.3.4.4. Molecular-genetic and virological analysis has revealed the differences between the viruses isolated in 2016–2017 and the virus of the same clade 2.3.4.4 that was isolated in 2014