Similarity analysis of silage, rumen and milk microbiota in dairy cows

Diseases that occur in such a multifactorial system as animal husbandry are determined not only by internal factors of the body, for example, the composition of the microflora of the digestive system, but also by external factors, such as feed. The aim of the study was to analyze the similarity of the microbiota of silage fed to cows with the composition of the microflora of their rumen and milk using the NGS sequencing method. The experiment was carried out on one of the commercial farms of the Leningrad region. The bacterial community of the contents of rumen, milk and silage from perennial cereals and legumes was evaluated by NGS sequencing on the MiSeq platform (Illumina, Inc., USA) with primers for the V3-V4 region of the 16S rRNA gene. As a result of the study, 22 phylum of attributed microorganisms were found in the microflora of silage, 24 to 30 phylum of rumen, 18 phylum of milk. Similar taxa of microorganisms were identified in silage, rumen and milk, the main difference was noted in quantitative ratios (P≤0.05). For example, the amount of Firmicutes in silage was 52.9 ± 3.45%, in milk - 11.8 ± 0.78%. This suggests that there may be some relationship between the studied biotopes. Pathogenic microorganisms, including the causative agents of mastitis, were present in many samples. The genera Staphylococcus, Acinetobacter, Streptococcus and Fusobacterium were identified by us as the most represented (P≤0.05) in the composition of the milk microflora. Their content was 0.24±0.023, 1.8±1.23, 1.0±0.06 and 0.35±0.031%, respectively. Probably, the microflora of the rumen can influence the formation of the milk microbiota

Myeloid-derived suppressor cells as biomarkers of the effectiveness of therapy with new biological agents in axial spondyloarthritis

Innate immune cells, including myeloid cells — myeloid derived suppressor cells (MDSCs) — are supposed to play an important role in the pathogenesis of axial spondyloarthritis (AxSp). Myeloid derived suppressor cells represent a heterogeneous population of immature cells capable of suppressing innate and adaptive immune responses with the most pronounced suppressor activity against T cells. Biological disease-modifying antirheumatic drugs (bDMARDs) can reduce the clinical and laboratory disease activity, but their effectiveness varies widely in different patients with AxSp. The present study is aimed at studying MDSCs subpopulations and their suppressive function depending on the response to bDMARD therapy in AxSp. The study included AxSp patients with a disease duration of 16.5 years (median); HLA-B27 (+) status was detected in 79% of cases. All patients received bDMARDs at least the past 12 weeks, including TNF inhibitors (etanercept, certolizumab pegol, adalimumab, or golimumab) or IL-17 inhibitors (secukinumab, ixekizumab, or netakimab). Percentage of granulocytic MDSCs (G-MDSCs, Lin-HLA-DR-CD33+CD66b+), monocytic MDSCs (M-MDSCs, HLA-DRlow/-CD14+), MDSCs of early stage differentiation (E-MDSCs, Lin-HLA-DR- CD33+CD66b-), as well as intracellular expression of arginase-1 was assessed by flow cytometry. Frequency of circulating MDSC subpopulations of patients with a stable response to bDMARDs (responders) did not differ significantly compared to healthy donors. Patients not responding to bDMARDs therapy showed increased relative and absolute number of E-MDSCs compared to healthy donors (pU = 0.01 and pU = 0.02, respectively) and the responders (pU = 0.03 and pU = 0.07, respectively). Increased percentage of E-MDSCs was positively correlated to disease activity — ESR (Rs = 0.821; p = 0.023), CRP (Rs = 0.714; p = 0.07) and ASDASCRP (Rs = 0.829; p = 0.042) in the non-responder group. Responder patients exhibited no correlation between disease activity and circulating MDSCs. The suppressor potential of MDSCs was analyzed by the intracellular expression of arginase-1 molecule which is involved in the inhibition of T cell response. Patients with the stable response were characterized by increased expression of arginase-1 in E-MDSCs compared to donors (pU = 0.02). Non-responders did not demonstrate significant changes in Arg-1 expression, however, the percentage of arginase-1-expressing G-MDSCs was positively correlated to indexes ASDASESR (Rs = 0.857; p = 0.014) and BASDAI (Rs = 0.785; p = 0.036). Thus, E-MDSCs as well as arginase-1 expression in MDSCs may serve as biomarkers of effectiveness bDMARD therapy, and act as potential candidate predictors of response to therapy in AxSp

Friedreich's ataxia-associated childhood hypertrophic cardiomyopathy: a national cohort study

OBJECTIVE: Hypertrophic cardiomyopathy (HCM) is an important predictor of long-term outcomes in Friedreich's ataxia (FA), but the clinical spectrum and survival in childhood is poorly described. This study aimed to describe the clinical characteristics of children with FA-HCM. DESIGN AND SETTING: Retrospective, longitudinal cohort study of children with FA-HCM from the UK. PATIENTS: 78 children (<18 years) with FA-HCM diagnosed over four decades. INTERVENTION: Anonymised retrospective demographic and clinical data were collected from baseline evaluation and follow-up. MAIN OUTCOME MEASURES: The primary study end-point was all-cause mortality (sudden cardiac death, atrial arrhythmia-related death, heart failure-related death, non-cardiac death) or cardiac transplantation. RESULTS: The mean age at diagnosis of FA-HCM was 10.9 (±3.1) years. Diagnosis was within 1 year of cardiac referral in 34 (65.0%) patients, but preceded the diagnosis of FA in 4 (5.3%). At baseline, 65 (90.3%) had concentric left ventricular hypertrophy and 6 (12.5%) had systolic impairment. Over a median follow-up of 5.1 years (IQR 2.4-7.3), 8 (10.5%) had documented supraventricular arrhythmias and 8 (10.5%) died (atrial arrhythmia-related n=2; heart failure-related n=1; non-cardiac n=2; or unknown cause n=3), but there were no sudden cardiac deaths. Freedom from death or transplantation at 10 years was 80.8% (95% CI 62.5 to 90.8). CONCLUSIONS: This is the largest cohort of childhood FA-HCM reported to date and describes a high prevalence of atrial arrhythmias and impaired systolic function in childhood, suggesting early progression to end-stage disease. Overall mortality is similar to that reported in non-syndromic childhood HCM, but no patients died suddenly

USAGE EFFICIENCY OF THE AIR AND THE DISPERSE ENVIRONMENT AS HEAT-CARRIERS

The estimation of the air and the disperse environment (fluidized bed) in the heat exchange intensity with identical energy expenses on a heat-carriers transportation is carried out in the work.В работе проведена оценка эффективности воздуха и дисперсной среды (псевдоожиженного слоя) по интенсивности теплообмена при одинаковых затратах энергии на перемещение теплоносителей

Studies of effector molecules exerting autonomous and nonautonomous influence of T lymphocyte apoptosis under the conditions of in vitro “cell neighborhood” in healthy people and patients with rheumatoid arthritis

Cellular homeostasis in the body is known to be maintained by the processes of cell proliferation and death, whereas apoptosis is the most frequent and physiological, “silent” mechanism of cell elimination. It has been currently shown that the process of apoptosis traditionally considered an autonomous event, has a pronounced non-autonomous effect on migration, proliferation, and death of the neighboring cells. This work was based on the data on impaired programmed death of mononuclear cells from the patients with rheumatoid arthritis (RA) leading to the evolving autoimmune inflammation. The aim of this study was to evaluate effector molecules exerting autonomous and non-autonomous influence of T cell apoptosis under the conditions of “cell neighborhood” in cell cultures of healthy people and RA patients. The studies were performed with blood samples of RA patients and healthy women of comparable age. These experiments were performed in order to assess the levels of main molecules mediating the in vitro receptor and mitochondrial apoptosis of T lymphocytes. In previous studies, using the original “cell neighborhood” model, no differences were found in parameters of early and late activation apoptosis between the groups of donors and RA patients. At the same time, 1-week incubation in apoptotic cultures of the patients was followed by significantly increased number of viable cells carrying the proliferation marker Ki-67. Different results of in vitro apoptosis induction in cultures under similar conditions of “cell neighborhood” in healthy people and patients with RA have revealed the importance of main effector molecules of apoptosis in the studied groups. In this study, we have revealed low potential of the receptor pathway for apoptosis activation in healthy people, due to suppression of TNFα production during cell incubation under the conditions of “cell neighborhood”, and in RA patients due to initially low TNFα in supernatants which did not change over time and in various incubation variants, along with low content of initiating caspase 8 in both groups. Significant suppression of effector molecules of mitochondrial pathway of apoptosis activation, i.e., Bcl-2 anti-apoptotic factor and p53 transcription factor was detected in cultures of apoptotic cells, as well as mixtures of proliferating and apoptotic cells under the conditions of “cell neighborhood” in RA patients. The amounts of these molecules did not change in healthy persons. At the same time, no differences in these molecules were found between individual variants of cell cultures from the patients with RA and healthy people. The both studied groups were characterized by a significant activation of IL-4 and IL-6 production, i.e., the cytokines with autonomous and non-autonomous protective and reparative properties, Hence, one may conclude that high levels of these cytokines had different effects in cell cultures under the conditions of “cell neighborhood”. Incubation of cells from healthy people under suboptimal conditions was associated with maintaining the balance of proliferation and apoptosis, whereas, in cell cultures of RA patients, this balance caused activation of proliferation processes, being accompanied by an increase in the number of living cells in apoptotic cultures

USE OF PIPES WITH KNURL WHEEL IN INSTALLATIONS OF FLASH EVAPORATOR

The paper considers the possibility of using developed surfaces in the form pipes with knurl in installations of flash evaporator. The effect of knurling on the heat exchange processes inherent in this installation is considered.В работе рассмотрена возможность использования развитых поверхностей в виде накатаных труб в установках мгновенного вскипания. Рассмотрено влияние накатки на процессы теплообмена, присущие этой установке

Comparison of the composition and metabolic potential of the reindeer’s rumen microbiome in the Yamal-Nenets and Nenets autonomous district of the Russian Arctic

The adaptive ability of reindeer to the harsh conditions of the Russian Arctic is not determined solely by the genome of the macroorganism and, of course, includes an extensive genetic and metabolic repertoire of the microbiome.The aim. To compare the taxonomic and predicted metabolic profiles of the rumen microbiome of adult reindeer living in the natural pastures of the Yamalo-Nenets and Nenets Autonomous districts of the Russian Federation.Materials and methods. Expeditions to the Yamal-Nenets and Nenets Autonomous districts of the Russian Arctic in 2017 were carried out to take samples of the rumen. The contents of the rumen were taken from clinically healthy reindeer individuals (at least 3 times repetition). To analyze the animal scar microbiota and determine metabolic profiles, 16S rRNA NGS sequencing was performed on a MiSeq device (Illumina, USA). Bioinformatic data analysis was performed using QIIME2 software ver. 2020.8. The noise sequences were filtered by DADA2. Silva 138 reference database was used for taxonomy analysis. Reconstruction and prediction of the functional content of the metagenome was carried out using the software complex PICRUSt2 (v. 2.3.0).Results. During NGS sequencing, a total of 223 768 sequences of the 16S rRNA gene of the reindeer scarring microbiome were studied. Significant (p ≤ 0.05) differences between the groups in 10 bacterial phyla and superphyla were revealed: Actinobacteriota, Spirochaetes, Chloroflexi, Verrucomicrobia, Bdellovibrionota, Synergistetes, Fusobacteriota, Myxococcota, Cyanobacteria, Campilobacterota. The results of the reconstruction and prediction of the functional content of the metagenome using the PICRUSt2 bioinformatic analysis made it possible to identify 328 potential metabolic pathways. Differences between the groups were revealed in 16 predicted metabolic pathways, among which the pathways of chlorophyllide and amino acid biosynthesis dominated

Data on gut metagenomes of the patients with Helicobacter pylori infection before and after the antibiotic therapy

© 2017Antibiotic therapy can lead to the disruption of gut microbiota community with possible negative outcomes for human health. One of the diseases for which the treatment scheme commonly included antibiotic intake is Helicobacter pylori infection. The changes in taxonomic and functional composition of microbiota in patients can be assessed using “shotgun” metagenomic sequencing. Ten stool samples were collected from 4 patients with Helicobacter pylori infection before and directly after the H. pylori eradication course. Additionally, for two of the subjects, the samples were collected 1 month after the end of the treatment. The samples were subject to “shotgun” (whole-genome) metagenomic sequencing using Illumina HiSeq platform. The reads are deposited in the ENA (project ID: PRJEB18265)

Sudden cardiac death in childhood RASopathy-associated hypertrophic cardiomyopathy: Validation of the HCM risk-kids model and predictors of events

Background: RASopathies account for nearly 20% of cases of childhood hypertrophic cardiomyopathy (HCM). Sudden cardiac death (SCD) occurs in patients with RASopathy-associated HCM, but the risk factors for SCD have not been systematically evaluated. Aim: To validate the HCM Risk-Kids SCD risk prediction model in children with RASopathy-associated HCM and investigate potential specific SCD predictors in this population. Methods: Validation of HCM Risk-Kids was performed in a retrospective cohort of 169 patients with a RASopathy-associated HCM from 15 international paediatric cardiology centres. Multiple imputation by chained equations was used for missing values related to the HCM Risk-Kids parameters. Results: Eleven patients (6.5%) experienced a SCD or equivalent event at a median age of 12.5 months (IQR 7.7–28.64). The calculated SCD/equivalent event incidence was 0.78 (95% CI 0.43–1.41) per 100 patient years. Six patients (54.54%) with an event were in the low-risk category according to the HCM Risk-Kids model. Harrell's C index was 0.60, with a sensitivity of 9.09%, specificity of 63.92%, positive predictive value of 1.72%, and negative predictive value of 91%; with a poor distinction between the different risk groups. Unexplained syncope (HR 42.17, 95% CI 10.49–169.56, p < 0.001) and non-sustained ventricular tachycardia (HR 5.48, 95% CI 1.58–19.03, p < 0.007) were predictors of SCD on univariate analysis. Conclusion: Unexplained syncope and the presence of NSVT emerge as predictors for SCD in children with RASopathy-associated HCM. The HCM Risk-Kids model may not be appropriate to use in this population, but larger multicentre collaborative studies are required to investigate this further

RNA-Seq gene expression profiling of HepG2 cells: The influence of experimental factors and comparison with liver tissue

© Tyakht et al.; licensee BioMed Central. Background: Human hepatoma HepG2 cells are used as an in vitro model of the human liver. High-throughput transcriptomic sequencing is an advanced approach for assessing the functional state of a tissue or cell type. However, the influence of experimental factors, such as the sample preparation method and inter-laboratory variation, on the transcriptomic profile has not been evaluated. Results: The whole-transcriptome sequencing of HepG2 cells was performed using the SOLiD platform and validated using droplet digital PCR. The gene expression profile was compared to the results obtained with the same sequencing method in another laboratory and using another sample preparation method. We also compared the transcriptomic profile HepG2 cells with that of liver tissue. Comparison of the gene expression profiles between the HepG2 cell line and liver tissue revealed the highest variation, followed by HepG2 cells submitted to two different sample preparation protocols. The lowest variation was observed between HepG2 cells prepared by two different laboratories using the same protocol. The enrichment analysis of the genes that were differentially expressed between HepG2 cells and liver tissue mainly revealed the cancer-associated gene signature of HepG2 cells and the activation of the response to chemical stimuli in the liver tissue. The HepG2 transcriptome obtained with the SOLiD platform was highly correlated with the published transcriptome obtained with the Illumina and Helicos platforms, with moderate correspondence to microarrays. Conclusions: In the present study, we assessed the influence of experimental factors on the HepG2 transcriptome and identified differences in gene expression between the HepG2 cell line and liver cells. These findings will facilitate robust experimental design in the fields of pharmacology and toxicology. Our results were supported by a comparative analysis with previous HepG2 gene expression studies