Определение концентрации холестерина и триглицеридов в экстраклеточных везикулах сыворотки крови с помощью коммерческих наборов

Exosomes and microvesicles, collectively referred to as small extracellular vesicles (sEV) are vesicles with an average size of about 100-150 nm. Currently, the role of sEV in various aspects of signaling in the body is being actively investigated; in addition, sEV can often serve as markers of various pathologies. The active study of the sEV composition is continuing. In this study we have demonstrated that in sEV it is possible to determine cholesterol and triglycerides concentration by using commercial kits designed for serum. The technique was tested on sEV from the blood of patients diagnosed with depression and on healthy volunteers. No differences were found in the concentration of cholesterol and triglycerides in mEV from the blood serum of depressed patients and the control group. The concentration of cholesterol and triglycerides in the samples is several times higher than the sensitivity threshold of the methods set by the manufacturer of the kits.Экзосомы и микровезикулы, совместно называемые малые экстраклеточные везикулы (мЭВ), представляют собой везикулы со средним размером около 100-150 нм. В настоящее время активно исследуется роль мЭВ в самых разных аспектах сигналинга в организме, кроме того, часто мЭВ могут служить маркерами разных патологий. Продолжается активное изучение состава мЭВ. В данной работе мы показали, что в мЭВ можно определить концентрацию холестерина и триглицеридов с помощью коммерческих наборов, предназначенных для сыворотки крови. Методика была опробована на мЭВ из крови пациентов с диагнозом депрессия и на здоровых добровольцах. Различий в концентрации холестерина и триглицеридов в мЭВ из сыворотки крови пациентов с депрессией и контрольной группы найдено не было. Концентрация холестерина и триглицеридов в образцах в несколько раз превосходит порог чувствительности методов, установленный производителем наборов

Neurotrophic factors in patients with primary open-angle glaucoma and age-related cataract. Part 2. Brain-derived neurotrophic factor

Purpose. To study the content of the brain-derived neurotrophic factor (BDNF) in the aqueous humor (AH), lacrimal fluid (LF), and blood serum (BS) in patients with primary open-angle glaucoma (POAG) and age-related cataract.Material and methods. We examined 141 people (141 eyes), of them 55 patients with POAG combined with age-related cataract, 57 patients with age-related cataract, and 29 persons without ophthalmic pathology. The content of BDNF in the AH (except healthy individuals), LF and BS was studied.Results. The levels of BDNF in LF and BS did not differ in cataract patients and healthy controls. Compared to patients with cataract the concentration of BDNF in patients with cataract and POAG was significantly reduced in the AH and LF (P<0.001), and in BS (P<0.05). In the early stage of POAG there was a significant decrease in the level of BDNF in all studied biological fluids (P<0.001), particularly pronounced in the AH – by more than 2.5 times. In subsequent stages, the BDNF levels consequentially increased in comparison with the early POAG, remaining, however, lower relative to the cataract patients (the difference was significant for the AH and LF, the correlation of the content of BDNF in the AH with the perimetric index VFI was negative: correlation coefficient r=-0.404, P=0.002). The level of BDNF in the AH showed a strong correlation with its concentration in the LF (r=0.66, P<0.000). A formula is proposed for an approximate calculation of the level of BDNF in the AH by its content in the LF.Conclusion. The content of BDNF in the AH, LF and BS is significantly reduced in patients with POAG, especially in its early stage. In subsequent stages, reduction of the level of BDNF in the AH and LF is consistently less pronounced, but remains decreased. A strong correlation of the content of BDNF in the AH with its concentration in the LF is established, which opens up new opportunities for indirect assessment of the level of BDNF in the AH of patients with POAG

THE SIGNIFICANCE OF OSTEOPONTINE AND MATRIX METALLOPROTEASE-9 IN THORACAL AORTA ANEURYSM DEVELOPMENT

Aim. To evaluate the significance of osteopontine and MMP-9 in the development of thoracal aorta aneurysm in patients with tricuspid (TAV) and bicuspid (BAV) aorta valve. Material and methods. Totally 94 patients included with the dilation of thoracal aorta for more than 40 mm, and 50 patients without aorta pathology, that were the comparison group. All patients underwent echocardiographic study by Vivid 7 (GE, USA) device by standard protocol. The osteopontine concentration and MMP-9 were measured in blood serum with manual plate immune-enzyme assay. Results. The concentration of MMP-9 in blood serum of the patients with aorta pathology and TAV did not differ significantly with the value in comparison group. Otherwise, in IHD the concentration of MMP-9 was significantly higher than in patients without aorta pathology (164,9±76,6 ng/ml and 106,8±82,7 ng/ml, respectively, p<0,01) and closely correlated with the Valsalva sinuses diameters (r=0,302, р=0,007). Comparative analysis of the osteopontine serum concentration did not show differences in the subgroups studied.Conclusion. The positive correlation revealed of the aorta diameter and concentration of MMP-9 in blood serum of the patients with BAV confirms not only the differences in the pathogenesis of thoracal aorta aneurysm in TAV and BAV, but makes it possible to think on the usage of MMP-9 as biomarker of thoracal aorta dilatation

Mapping cisplatin-induced viscosity alterations in cancer cells using molecular rotor and fluorescence lifetime imaging microscopy

Significance: Despite the importance of the cell membrane in regulation of drug activity, the influence of drug treatments on its physical properties is still poorly understood. The combination of fluorescence lifetime imaging microscopy (FLIM) with specific viscosity-sensitive fluorescent molecular rotors allows the quantification of membrane viscosity with high spatiotemporal resolution, down to the individual cell organelles. Aim: The aim of our work was to analyze microviscosity of the plasma membrane of living cancer cells during chemotherapy with cisplatin using FLIM and correlate the observed changes with lipid composition and cell’s response to treatment. Approach: FLIM together with viscosity-sensitive boron dipyrromethene-based fluorescent molecular rotor was used to map the fluidity of the cell’s membrane. Chemical analysis of membrane lipid composition was performed with time-of-flight secondary ion mass spectrometry (ToF-SIMS). Results: We detected a significant steady increase in membrane viscosity in viable cancer cells, both in cell monolayers and tumor spheroids, upon prolonged treatment with cisplatin, as well as in cisplatin-adapted cell line. ToF-SIMS revealed correlative changes in lipid profile of cisplatin-treated cells. Conclusions: These results suggest an involvement of membrane viscosity in the cell adaptation to the drug and in the acquisition of drug resistance

The contribution of rat studies to contemporary knowledge of Major Depressive Disorder: Results from citation analysis

Funding: This study was financed by Animalfree Research—Switzerland, a grant from the Johns Hopkins Center for Alternatives to Animal Testing (CAAT) and by Portuguese National Funds through FCT—Fundação para a Ciência e a Tecnologia, within the CFCUL Unit funding UIDB/00678/2020. TM thanks partial support by CEAUL (funded by FCT—Fundação para a Ciência e a Tecnologia, Portugal, through the project UID/MAT/00006/2019). FP thanks FCT/MCTES for the financial support to CESAM (UIDP/50017/2020 and UIDB/50017/2020), through national funds. Open access publication costs were covered by Animalfree Research-Switzerland and by funding provided by the Ketty and Leif Hjordt Foundation.Major depressive disorder (MDD) is the most severe depression type and one of the leading causes of morbidity worldwide. Animal models are widely used to understand MDD etiology, pathogenesis, and treatment, but the efficacy of this research for patients has barely been systematically evaluated. Such evaluation is important given the resource consumption and ethical concerns incurred by animal use. We used the citation tracking facilities within Web of Science and Scopus to locate citations of original research papers on rats related to MDD published prior to 2013—to allow adequate time for citations—identified in PubMed and Scopus by relevant search terms. Resulting citations were thematically coded in eight categories, and descriptive statistics were calculated. 178 publications describing relevant rat studies were identified. They were cited 8,712 times. More than half (4,633) of their citations were by other animal studies. 794 (less than 10%) were by human medical papers. Citation analysis indicates that rat model research has contributed very little to the contemporary clinical understanding of MDD. This suggests a misuse of limited funding hence supporting a change in allocation of research and development funds targeting this disorder to maximise benefits for patients.Publisher PDFPeer reviewe

Low-pass single-chromosome sequencing of human small supernumerary marker chromosomes (sSMCs) and Apodemus B chromosomes.

Supernumerary chromosomes sporadically arise in many eukaryotic species as a result of genomic rearrangements. If present in a substantial part of species population, those are called B chromosomes, or Bs. This is the case for 70 mammalian species, most of which are rodents. In humans, the most common types of extra chromosomes, sSMCs (small supernumerary marker chromosomes), are diagnosed in approximately 1 of 2000 postnatal cases. Due to low frequency in population, human sSMCs are not considered B chromosomes. Genetic content of both B-chromosomes and sSMCs in most cases remains understudied. Here, we apply microdissection of single chromosomes with subsequent low-pass sequencing on Ion Torrent PGM and Illumina MiSeq to identify unique and repetitive DNA sequences present in a single human sSMC and several B chromosomes in mice Apodemus flavicollis and Apodemus peninsulae. The pipeline for sequencing data analysis was made available in Galaxy interface as an addition to previously published command-line version. Human sSMC was attributed to the proximal part of chromosome 15 long arm, and breakpoints leading to its formation were located into satellite DNA arrays. Genetic content of Apodemus B chromosomes was species-specific, and minor alterations were observed in both species. Common features of Bs in these Apodemus species were satellite DNA and ERV enrichment, as well as the presence of the vaccinia-related kinase gene Vrk1. Understanding of the non-essential genome elements content provides important insights into genome evolution in general.This is a post-peer-review, pre-copyedit version of an article published in Chromosoma. The final authenticated version is available online at: [http://dx.doi.org/10.1007/s00412-018-0662-0

Genotyping of Capreolus pygargus Fossil DNA from Denisova Cave Reveals Phylogenetic Relationships between Ancient and Modern Populations

BACKGROUND: The extant roe deer (Capreolus Gray, 1821) includes two species: the European roe deer (C. capreolus) and the Siberian roe deer (C. pygargus) that are distinguished by morphological and karyotypical differences. The Siberian roe deer occupies a vast area of Asia and is considerably less studied than the European roe deer. Modern systematics of the Siberian roe deer remain controversial with 4 morphological subspecies. Roe deer fossilized bones are quite abundant in Denisova cave (Altai Mountains, South Siberia), where dozens of both extant and extinct mammalian species from modern Holocene to Middle Pleistocene have been retrieved. METHODOLOGY/PRINCIPAL FINDINGS: We analyzed a 629 bp fragment of the mitochondrial control region from ancient bones of 10 Holocene and four Pleistocene Siberian roe deer from Denisova cave as well as 37 modern specimen belonging to populations from Altai, Tian Shan (Kyrgyzstan), Yakutia, Novosibirsk region and the Russian Far East. Genealogical reconstructions indicated that most Holocene haplotypes were probably ancestral for modern roe deer populations of Western Siberia and Tian Shan. One of the Pleistocene haplotypes was possibly ancestral for modern Yakutian populations, and two extinct Pleistocene haplotypes were close to modern roe deer from Tian Shan and Yakutia. Most modern geographical populations (except for West Siberian Plains) are heterogeneous and there is some tentative evidence for structure. However, we did not find any distinct phylogenetic signal characterizing particular subspecies in either modern or ancient samples. CONCLUSION/SIGNIFICANCE: Analysis of mitochondrial DNA from both ancient and modern samples of Siberian roe deer shed new light on understanding the evolutionary history of roe deer. Our data indicate that during the last 50,000 years multiple replacements of populations of the Siberian roe deer took place in the Altai Mountains correlating with climatic changes. The Siberian roe deer represent a complex and heterogeneous species with high migration rates and without evident subspecies structure. Low genetic diversity of the West Siberian Plain population indicates a recent bottleneck or founder effect