Identification of African Swine Fever Virus Transcription within Peripheral Blood Mononuclear Cells of Acutely Infected Pigs

African swine fever virus (ASFV) has become widespread in Europe, Asia and elsewhere, thereby causing extensive economic losses. The viral genome includes nearly 200 genes, but their expression within infected pigs has not been well characterized previously. In this study, four pigs were infected with a genotype II strain (ASFV POL/2015/Podlaskie); blood samples were collected before inoculation and at both 3 and 6 days later. During this period, a range of clinical signs of infection became apparent in the pigs. From the blood, peripheral blood mononuclear cells (PBMCs) were isolated. The transcription of the ASFV genes was determined using RNAseq on poly(A)+ mRNAs isolated from these cells. Only very low levels of virus transcription were detected in the PBMCs at 3 days post-inoculation (dpi) but, at 6 dpi, extensive transcription was apparent. This was co-incident with a large increase in the level of ASFV DNA within these cells. The pattern of the virus gene expression was very reproducible between the individual pigs. Many highly expressed genes have undefined roles. Surprisingly, some genes with key roles in virus replication were expressed at only low levels. As the functions of individual genes are identified, information about their expression becomes important for understanding their contribution to virus biology.info:eu-repo/semantics/publishedVersio

Identification of African Swine Fever Virus Transcription within Peripheral Blood Mononuclear Cells of Acutely Infected Pigs

African swine fever virus (ASFV) has become widespread in Europe, Asia and elsewhere, thereby causing extensive economic losses. The viral genome includes nearly 200 genes, but their expression within infected pigs has not been well characterized previously. In this study, four pigs were infected with a genotype II strain (ASFV POL/2015/Podlaskie); blood samples were collected before inoculation and at both 3 and 6 days later. During this period, a range of clinical signs of infection became apparent in the pigs. From the blood, peripheral blood mononuclear cells (PBMCs) were isolated. The transcription of the ASFV genes was determined using RNAseq on poly(A)+ mRNAs isolated from these cells. Only very low levels of virus transcription were detected in the PBMCs at 3 days post-inoculation (dpi) but, at 6 dpi, extensive transcription was apparent. This was co-incident with a large increase in the level of ASFV DNA within these cells. The pattern of the virus gene expression was very reproducible between the individual pigs. Many highly expressed genes have undefined roles. Surprisingly, some genes with key roles in virus replication were expressed at only low levels. As the functions of individual genes are identified, information about their expression becomes important for understanding their contribution to virus biology

Анализ наночастиц и наноматериалов методом рентгеновской фотоэлектронной спектроскопии

Objectives. The main aim of this review is to summarize the existing knowledge on the use of X-ray photoelectron spectroscopy (XPS) for the characterization of nanoparticles and nanomaterials.Results. XPS or electron spectroscopy for chemical analysis can provide information on the qualitative and quantitative composition, valence states of the elements of the samples under study, the chemical composition of the surface and interfaces that determine the properties of nanoparticles and nanostructured materials. The review describes the role of several different methods for the characterization of nanomaterials, highlights their advantages and limitations, and the possibilities of an effective combination. The main characteristics of XPS are described. Various examples of its use for the analysis of nanoparticles and nanomaterials are given in conjunction with additional methods to obtain complementary information about the object under study.Conclusions. XPS provides depth information comparable to the size of nanoparticles (up to 10 nm depth from the surface) and does not cause significant damage to the samples. Two disadvantages of XPS analysis are sample preparation requiring a dry solid form without contaminations and data interpretation. XPS provides information not only on the chemical identity, but also on the dielectric properties of nanomaterials, recording their charging/discharging behavior. Chemical information from the surface of nanoparticles analyzed by XPS can be used to estimate the thickness of nanoparticle coatings. XPS has a high selectivity, since the resolution of the method makes it possible to distinguish a characteristic set of lines in the photoelectron spectrum at kinetic energies determined by the photon energy and the corresponding binding energies in elements. The intensity of the lines depends on the concentration of the respective element. Obtaining a sufficiently complete picture of the properties of nanomaterials requires the use of a group of complementary instrumental methods of analysis.Цели. Основная цель данного обзора - обобщить существующие знания об использовании метода рентгеновской фотоэлектронной спектроскопии (РФЭС) для характеризации наночастиц и наноматериалов.Результаты. Метод РФЭС или электронной спектроскопии для химического анализа может предоставить информацию о качественном и количественном составе, валентных состояниях элементов исследуемых образцов, химическом составе поверхности и границ раздела, которые определяют свойства наночастиц и наноструктурных материалов. В обзоре описана роль нескольких различных методов для характеристики наноразмерных материалов, подчеркнуты их преимущества, ограничения и возможности эффективной комбинации. Описаны основные характеристики РФЭС. Приведены различные примеры ее использования для анализа наночастиц и наноматериалов в совокупности с дополнительными методами для получения комплементарной информации об изучаемом объекте.Выводы. РФЭС предоставляет информацию о глубине, сравнимой с размером наночастиц (до 10 нм глубины от поверхности), и не вызывает значительного повреждения образцов. Двумя недостатками анализа РФЭС являются подготовка образцов (требуется сухая твердая форма без загрязнения) и интерпретация данных. РФЭС предоставляет информацию не только о химической идентичности, но и о диэлектрических свойствах наноматериалов, регистрируя их поведение при зарядке/разрядке. Химическая информация с поверхности наночастиц, проанализированная с помощью РФЭС, может использоваться для оценки толщины покрытий наночастиц. РФЭС обладает высокой селективностью, поскольку разрешающая способность метода позволяет различить характерный набор линий в фотоэлектронном спектре при кинетических энергиях, определяемых энергией фотонов и соответствующими энергиями связи в элементах. Интенсивность линий зависит от концентрации соответствующего элемента. Получение достаточно полной картины свойств наноматериалов требует использования группы взаимодополняющих инструментальных методов анализа

Характеризация наночастиц кристаллического кремния, легированного железом, и их модификация цитрат-анионами для использования in vivo

Objectives. This paper presents data on the development and study of the structural properties of iron-doped crystalline silicon (nc-Si/SiOx/Fe) nanoparticles obtained using the plasma-chemical method for application in magnetic resonance imaging diagnostics and treatment of oncological diseases. This work aimed to use a variety of analytical methods to study the structural properties of nc-Si/SiOx/Fe and their colloidal stabilization with citrate anions for in vivo applications.Methods. Silicon nanoparticles obtained via the plasma-chemical synthesis method were characterized by laser spark emission spectroscopy, atomic emission spectroscopy, Fouriertransform infrared spectroscopy, and X-ray photoelectron spectroscopy. The hydrodynamic diameter of the nanoparticles was estimated using dynamic light scattering. The toxicity of the nanoparticles was investigated using a colorimetric MTT test for the cell metabolic activity. Elemental iron with different Fe/Si atomic ratios was added to the feedstock during loading.Results. The particles were shown to have a large silicon core covered by a relatively thin layer of intermediate oxides (interface) and an amorphous oxide shell, which is silicon oxide with different oxidation states SiOx (0 ≤ x ≤ 2). The samples had an iron content of 0.8–1.8 at %. Colloidal solutions of the nanoparticles stabilized by citrate anions were obtained and characterized. According to the analysis of the cytotoxicity of the modified nanosilicon particles using monoclonal K562 human erythroleukemia cells, no toxicity was found for cells in culture at particle concentrations of up to 5 µg/mL.Conclusions. Since the obtained modified particles are nontoxic, they can be used in in vivo theranostic applications.Цели. В работе приводятся данные по разработке и изучению структурных свойств полученных плазмохимическим методом наночастиц кремния nc-Si/SiOx/Fe, легированных железом. Цель работы – исследование свойств наночастиц кремния, легированных железом, комплексом аналитических методов и их стабилизация цитрат-анионами для применения в диагностике методом магнитно-резонансной томографии и лечении онкологических заболеваний.Методы. Наночастицы кремния, полученные плазмохимическим методом синтеза, были охарактеризованы лазерно-искровым эмиссионным методом, методом атомной эмиссионной спектроскопии, Фурье-ИК-спектроскопией, рентгеновской фотоэлектронной спектроскопией. Гидродинамический диаметр наночастиц оценивали методом динамического светорассеяния. Исследование токсичности наночастиц проводили с помощью колориметрического МТТ теста на метаболическую активность клеток. В исходное сырье при загрузке добавляли элементарное железо с разным атомным соотношением Fe/Si.Результаты. Было показано, что частица имеет кремниевое ядро с аморфной оксидной оболочкой, представляющей собой оксиды кремния с разной степенью окисления SiO x (0 ≤ x ≤ 2). Содержание железа в образцах составило от 0.8 до 1.8 ат. %. Были получены и охарактеризованы коллоидные растворы наночастиц, стабилизированные цитрат-анионами. Анализ цитотоксичности модифицированных частиц нанокремния с использованием моноклонизированных клеток эритролейкоза человека К562 показал отсутствие токсичности для клеток в культуре при концентрации частиц до 5 мкг/мл.Выводы. Полученные модифицированные частицы не обладают токсичностью, поэтому их можно рекомендовать для использования в in vivo приложениях для тераностик

Antiinflammatory Therapy with Canakinumab for Atherosclerotic Disease

Background: Experimental and clinical data suggest that reducing inflammation without affecting lipid levels may reduce the risk of cardiovascular disease. Yet, the inflammatory hypothesis of atherothrombosis has remained unproved. Methods: We conducted a randomized, double-blind trial of canakinumab, a therapeutic monoclonal antibody targeting interleukin-1β, involving 10,061 patients with previous myocardial infarction and a high-sensitivity C-reactive protein level of 2 mg or more per liter. The trial compared three doses of canakinumab (50 mg, 150 mg, and 300 mg, administered subcutaneously every 3 months) with placebo. The primary efficacy end point was nonfatal myocardial infarction, nonfatal stroke, or cardiovascular death. RESULTS: At 48 months, the median reduction from baseline in the high-sensitivity C-reactive protein level was 26 percentage points greater in the group that received the 50-mg dose of canakinumab, 37 percentage points greater in the 150-mg group, and 41 percentage points greater in the 300-mg group than in the placebo group. Canakinumab did not reduce lipid levels from baseline. At a median follow-up of 3.7 years, the incidence rate for the primary end point was 4.50 events per 100 person-years in the placebo group, 4.11 events per 100 person-years in the 50-mg group, 3.86 events per 100 person-years in the 150-mg group, and 3.90 events per 100 person-years in the 300-mg group. The hazard ratios as compared with placebo were as follows: in the 50-mg group, 0.93 (95% confidence interval [CI], 0.80 to 1.07; P = 0.30); in the 150-mg group, 0.85 (95% CI, 0.74 to 0.98; P = 0.021); and in the 300-mg group, 0.86 (95% CI, 0.75 to 0.99; P = 0.031). The 150-mg dose, but not the other doses, met the prespecified multiplicity-adjusted threshold for statistical significance for the primary end point and the secondary end point that additionally included hospitalization for unstable angina that led to urgent revascularization (hazard ratio vs. placebo, 0.83; 95% CI, 0.73 to 0.95; P = 0.005). Canakinumab was associated with a higher incidence of fatal infection than was placebo. There was no significant difference in all-cause mortality (hazard ratio for all canakinumab doses vs. placebo, 0.94; 95% CI, 0.83 to 1.06; P = 0.31). Conclusions: Antiinflammatory therapy targeting the interleukin-1β innate immunity pathway with canakinumab at a dose of 150 mg every 3 months led to a significantly lower rate of recurrent cardiovascular events than placebo, independent of lipid-level lowering. (Funded by Novartis; CANTOS ClinicalTrials.gov number, NCT01327846.

Effects of alirocumab on types of myocardial infarction: insights from the ODYSSEY OUTCOMES trial

Aims  The third Universal Definition of Myocardial Infarction (MI) Task Force classified MIs into five types: Type 1, spontaneous; Type 2, related to oxygen supply/demand imbalance; Type 3, fatal without ascertainment of cardiac biomarkers; Type 4, related to percutaneous coronary intervention; and Type 5, related to coronary artery bypass surgery. Low-density lipoprotein cholesterol (LDL-C) reduction with statins and proprotein convertase subtilisin–kexin Type 9 (PCSK9) inhibitors reduces risk of MI, but less is known about effects on types of MI. ODYSSEY OUTCOMES compared the PCSK9 inhibitor alirocumab with placebo in 18 924 patients with recent acute coronary syndrome (ACS) and elevated LDL-C (≥1.8 mmol/L) despite intensive statin therapy. In a pre-specified analysis, we assessed the effects of alirocumab on types of MI. Methods and results  Median follow-up was 2.8 years. Myocardial infarction types were prospectively adjudicated and classified. Of 1860 total MIs, 1223 (65.8%) were adjudicated as Type 1, 386 (20.8%) as Type 2, and 244 (13.1%) as Type 4. Few events were Type 3 (n = 2) or Type 5 (n = 5). Alirocumab reduced first MIs [hazard ratio (HR) 0.85, 95% confidence interval (CI) 0.77–0.95; P = 0.003], with reductions in both Type 1 (HR 0.87, 95% CI 0.77–0.99; P = 0.032) and Type 2 (0.77, 0.61–0.97; P = 0.025), but not Type 4 MI. Conclusion  After ACS, alirocumab added to intensive statin therapy favourably impacted on Type 1 and 2 MIs. The data indicate for the first time that a lipid-lowering therapy can attenuate the risk of Type 2 MI. Low-density lipoprotein cholesterol reduction below levels achievable with statins is an effective preventive strategy for both MI types.For complete list of authors see http://dx.doi.org/10.1093/eurheartj/ehz299</p

Evacetrapib and Cardiovascular Outcomes in High-Risk Vascular Disease

BACKGROUND: The cholesteryl ester transfer protein inhibitor evacetrapib substantially raises the high-density lipoprotein (HDL) cholesterol level, reduces the low-density lipoprotein (LDL) cholesterol level, and enhances cellular cholesterol efflux capacity. We sought to determine the effect of evacetrapib on major adverse cardiovascular outcomes in patients with high-risk vascular disease. METHODS: In a multicenter, randomized, double-blind, placebo-controlled phase 3 trial, we enrolled 12,092 patients who had at least one of the following conditions: an acute coronary syndrome within the previous 30 to 365 days, cerebrovascular atherosclerotic disease, peripheral vascular arterial disease, or diabetes mellitus with coronary artery disease. Patients were randomly assigned to receive either evacetrapib at a dose of 130 mg or matching placebo, administered daily, in addition to standard medical therapy. The primary efficacy end point was the first occurrence of any component of the composite of death from cardiovascular causes, myocardial infarction, stroke, coronary revascularization, or hospitalization for unstable angina. RESULTS: At 3 months, a 31.1% decrease in the mean LDL cholesterol level was observed with evacetrapib versus a 6.0% increase with placebo, and a 133.2% increase in the mean HDL cholesterol level was seen with evacetrapib versus a 1.6% increase with placebo. After 1363 of the planned 1670 primary end-point events had occurred, the data and safety monitoring board recommended that the trial be terminated early because of a lack of efficacy. After a median of 26 months of evacetrapib or placebo, a primary end-point event occurred in 12.9% of the patients in the evacetrapib group and in 12.8% of those in the placebo group (hazard ratio, 1.01; 95% confidence interval, 0.91 to 1.11; P=0.91). CONCLUSIONS: Although the cholesteryl ester transfer protein inhibitor evacetrapib had favorable effects on established lipid biomarkers, treatment with evacetrapib did not result in a lower rate of cardiovascular events than placebo among patients with high-risk vascular disease. (Funded by Eli Lilly; ACCELERATE ClinicalTrials.gov number, NCT01687998 .)

Effect of alirocumab on mortality after acute coronary syndromes. An analysis of the ODYSSEY OUTCOMES randomized clinical trial

Background: Previous trials of PCSK9 (proprotein convertase subtilisin-kexin type 9) inhibitors demonstrated reductions in major adverse cardiovascular events, but not death. We assessed the effects of alirocumab on death after index acute coronary syndrome. Methods: ODYSSEY OUTCOMES (Evaluation of Cardiovascular Outcomes After an Acute Coronary Syndrome During Treatment With Alirocumab) was a double-blind, randomized comparison of alirocumab or placebo in 18 924 patients who had an ACS 1 to 12 months previously and elevated atherogenic lipoproteins despite intensive statin therapy. Alirocumab dose was blindly titrated to target achieved low-density lipoprotein cholesterol (LDL-C) between 25 and 50 mg/dL. We examined the effects of treatment on all-cause death and its components, cardiovascular and noncardiovascular death, with log-rank testing. Joint semiparametric models tested associations between nonfatal cardiovascular events and cardiovascular or noncardiovascular death. Results: Median follow-up was 2.8 years. Death occurred in 334 (3.5%) and 392 (4.1%) patients, respectively, in the alirocumab and placebo groups (hazard ratio [HR], 0.85; 95% CI, 0.73 to 0.98; P=0.03, nominal P value). This resulted from nonsignificantly fewer cardiovascular (240 [2.5%] vs 271 [2.9%]; HR, 0.88; 95% CI, 0.74 to 1.05; P=0.15) and noncardiovascular (94 [1.0%] vs 121 [1.3%]; HR, 0.77; 95% CI, 0.59 to 1.01; P=0.06) deaths with alirocumab. In a prespecified analysis of 8242 patients eligible for ≥3 years follow-up, alirocumab reduced death (HR, 0.78; 95% CI, 0.65 to 0.94; P=0.01). Patients with nonfatal cardiovascular events were at increased risk for cardiovascular and noncardiovascular deaths (P<0.0001 for the associations). Alirocumab reduced total nonfatal cardiovascular events (P<0.001) and thereby may have attenuated the number of cardiovascular and noncardiovascular deaths. A post hoc analysis found that, compared to patients with lower LDL-C, patients with baseline LDL-C ≥100 mg/dL (2.59 mmol/L) had a greater absolute risk of death and a larger mortality benefit from alirocumab (HR, 0.71; 95% CI, 0.56 to 0.90; Pinteraction=0.007). In the alirocumab group, all-cause death declined wit h achieved LDL-C at 4 months of treatment, to a level of approximately 30 mg/dL (adjusted P=0.017 for linear trend). Conclusions: Alirocumab added to intensive statin therapy has the potential to reduce death after acute coronary syndrome, particularly if treatment is maintained for ≥3 years, if baseline LDL-C is ≥100 mg/dL, or if achieved LDL-C is low. Clinical Trial Registration: URL: https://www.clinicaltrials.gov. Unique identifier: NCT01663402