The Nature-Based Ecological Engineering Paradigm: Symbiosis, Coupling, and Coordination

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A novel method for purifying bluetongue virus with high purity by co-immunoprecipitation with agarose protein A

<p>Abstract</p> <p>Background</p> <p>Bluetongue virus (BTV) is an icosahedral non-enveloped virus within the genus <it>Orbivirus </it>of <it>Reoviridae </it>and exists as 24 distinct serotypes. BTV can infect all ruminant species and causes severe sickness in sheep. Recently, it was reported that BTV can infect some human cancer cells selectively. Because of the important oncolysis of this virus, we developed a novel purifying method for large-scale production. The purifying logic is simple, which is picking out all the components unwanted and the left is what we want. The process can be summarized in 4 steps: centrifugation, pulling down cell debrises and soluble proteins by co-immunoprecipitation with agarose Protein A, dialysis and filtration sterilization after concentration.</p> <p>Results</p> <p>The result of transmission electron microscope (TEM) observation showed that the sample of purified virus has a very clear background and the virions still kept intact. The result of 50% tissue culture infective dose (TCID₅₀) assay showed that the bioactivity of purified virus is relatively high.</p> <p>Conclusions</p> <p>This method can purify BTV-10 with high quality and high biological activity on large-scale production. It also can be used for purifying other BTV serotypes.</p

PO-034 Effect of Voluntary Exercise on Cartilage Morphology of Knee Osteoarthritis in Obese Mice Induced by High-fat Diet

Objective To examine the effect of voluntary wheel-running exercise on cartilage morphology of knee osteoarthritis(KOA) in obese mice induced by high-fat diet,and explore the protective role of 4 weeks voluntary wheel-running exercise on KOA,finally providing effective experimental evidence for clinical treatment of knee osteoarthritis. Methods C57BL/6J mice were randomly assigned to the C-Sed group,C-Ex group,HF-Sed group and HF-Ex group.The control groups were fed a control diet(13.5% kcal from fat),and the high-fat groups were fed a high-fat diet(60% kcal from fat).After feeding 8 weeks different diets,the exercise groups were starting running.In order to examine the effect of voluntary wheel-running exercise on cartilage morphology of KOA,the joint of knee were harvested to be fixed,decalcified and embedded in paraffin,and the four-micrometer-thick sections were stained with both HE and toluidine blue . Results After feeding twelve weeks different diets,the body mass of the high-fat diet group mice has a significant increase,which demonstrates that high-fat diet could successfully induce the mice obese.From the results of HE and toluidine blue,in comparison to the C-Sed group,the surface of the knee articular cartilage in the HF-Sed group was not intact and smooth,and the thickness of articular cartilage has a significant decrease（p&lt;0.001）;contrary to the HF-Sed group,the surface of the knee articular cartilage in HF-Ex group was slightly smooth,and there was significant increase in cartilage thickness. Conclusions Four weeks voluntary wheel-running exercise can increase cartilage thickness ,decrease the Mankin’s score and delay the degeneration of knee cartilage in obese mice.To conclude,the short-term wheel-running exercise protects against obesity-induced KOA

Nitrogen availability does not affect ozone flux-effect relationships for biomass in birch (Betula pendula) saplings

To investigate whether nitrogen (N) load affects the ozone (O3) stomatal flux-effect relationship for birch biomass, three-year old birch saplings were exposed to seven different O3 profiles (24 h mean of 35–66 ppb) and four different N loads (10, 30, 50 and 70 kg ha−1 yr−1) in precision-controlled hemispherical glasshouses (solardomes) in 2012 and 2013. Stomatal conductance (gs) under optimal growth conditions was stimulated by enhanced N supply but was not significantly affected by enhanced O3 exposure. Birch root, woody (stem + branches) and total biomass (root + woody) were not affected by the Phytotoxic Ozone Dose (POD1SPEC) after two seasons of O3 exposure, and enhanced N supply stimulated biomass production independent of POD1SPEC (i.e. there were no POD1SPEC × N interactions). There was a strong linear relationship between the stem cross-sectional area and tree biomass at the end of the experiment, which was not affected by O3 exposure or N load. Enhanced N supply stimulated the stem cross-sectional area at the end of season 2, but not at the end of season 1, which suggests a time lag before tree biomass responded to enhanced N supply. There was no significant effect of POD1SPEC on stem cross-sectional area after either the first or second growing season of the experiment. Contrasting results reported in the literature on the interactive impacts of O3 and N load on tree physiology and growth are likely due to species-specific responses, different duration of the experiments and/or a limitation of the number of O3 and N levels tested

Single Cells Are Spatial Tokens: Transformers for Spatial Transcriptomic Data Imputation

Spatially resolved transcriptomics brings exciting breakthroughs to single-cell analysis by providing physical locations along with gene expression. However, as a cost of the extremely high spatial resolution, the cellular level spatial transcriptomic data suffer significantly from missing values. While a standard solution is to perform imputation on the missing values, most existing methods either overlook spatial information or only incorporate localized spatial context without the ability to capture long-range spatial information. Using multi-head self-attention mechanisms and positional encoding, transformer models can readily grasp the relationship between tokens and encode location information. In this paper, by treating single cells as spatial tokens, we study how to leverage transformers to facilitate spatial tanscriptomics imputation. In particular, investigate the following two key questions: (1) $\textit{how to encode spatial information of cells in transformers}$, and (2) $\textit{ how to train a transformer for transcriptomic imputation}$. By answering these two questions, we present a transformer-based imputation framework, SpaFormer, for cellular-level spatial transcriptomic data. Extensive experiments demonstrate that SpaFormer outperforms existing state-of-the-art imputation algorithms on three large-scale datasets while maintaining superior computational efficiency

OR-024 Changes of mitochondrial autophagy - related genes and autophagosome after skeletal muscle blunt trauma

Objective Objective: To study the changes of mitochondrial autophagy-related genes and autophagosome after skeletal muscle blunt trauma, to reveal the changes of mitochondrial adaptive repair process after skeletal muscle blunt trauma, and to elucidate the mechanism of blunt trauma repair process. Methods Methods: Sixty - four male Wistar rats were randomly divided into control group and blunt trauma group三 (divided into 12h group, 2d group, 5d group, 7d group, 10d group, 15d group and 30d group) according to the time of extraction. The expression of HIF-1α, AMPKα2, BNIP3 and NIX protein in skeletal muscle hypoxia and autophagy-related factors were measured by Western-Blot. QRT-PCR was performed to analyze the expression levels of HIF-1α, AMPKα2, BNIP3 and NIX. The ultrastructure and autophagic formation at different time points were observed by transmission electron microscopy (TEM). Results Results: The expression of HIF-1α and AMPKα2 protein reached the peak at 12h and 2d, and the expression of HIF-1α was significantly higher than that of the control group (P &lt;0.05). The expression of AMPKα2 was significantly higher at 5 days after injury (P &lt;0.05), and reached the normal level at 10 days. BNIP3 began to decline after 5 days, but still higher than normal at 10 days after treatment. NIX expression peak appeared at 12h and 2d after injury, with high-express to 7d. The expression of HIF-1α and AMPKα2 mRNA was significantly higher than that of the control group (P &lt;0.01), but decreased until 5d (P &lt;0.05), then decreased to normal level. The mRNA expression of BNIP3 and NIX was basically the same as their protein performance. A number of autophagosomes were observed at 12 h after injury, and the number of autophagosomes increased gradually at 2-7 d. After 10 days, the number of autophagosomes decreased compared with that of 12 h-7 d after blunt. And after 15 days, the number of autophagosites decreased gradually. Conclusions Conclusion: The changes of early stage metabolic regulator AMPKα2 and hypoxia-sensitive factor HIF-1αafter skeletal muscle blunt trauma indicated that&nbsp; an energy crisis occurred in the skeletal muscle after injury, and the hypoxic environment was formed. The mitochondrial autophagy, the expression of BNIP3 and NIX showed that mitochondrial autophagy was activated and hypoxia induced mitochondrial autophagy at early skeletal muscle contusion peroid. Hypoxia-induced mitochondrial autophagy could remove the damaged mitochondria, maintain mitochondrial quality and provide raw materials for new mitochondria generation, facilitate the rapid recovery of damaged skeletal muscle, which may be a compensatory mechanism of the body response to injury

Jump-seq: Genome-Wide Capture and Amplification of 5-Hydroxymethylcytosine Sites

5-Hydroxymethylcytosine (5hmC) arises from the oxidation of 5-methylcytosine (5mC) by Fe2+ and 2-oxoglutarate-dependent 10–11 translocation (TET) family proteins. Substantial levels of 5hmC accumulate in many mammalian tissues, especially in neurons and embryonic stem cells, suggesting a potential active role for 5hmC in epigenetic regulation beyond being simply an intermediate of active DNA demethylation. 5mC and 5hmC undergo dynamic changes during embryogenesis, neurogenesis, hematopoietic development, and oncogenesis. While methods have been developed to map 5hmC, more efficient approaches to detect 5hmC at base resolution are still highly desirable. Herein, we present a new method, Jump-seq, to capture and amplify 5hmC in genomic DNA. The principle of this method is to label 5hmC by the 6-N3-glucose moiety and connect a hairpin DNA oligonucleotide carrying an alkyne group to the azide-modified 5hmC via Huisgen cycloaddition (click) chemistry. Primer extension starts from the hairpin motif to the modified 5hmC site and then continues to “land” on genomic DNA. 5hmC sites are inferred from genomic DNA sequences immediately spanning the 5-prime junction. This technology was validated, and its utility in 5hmC identification was confirmed

Correlation analysis between 18F-fluorodeoxyglucose positron emission tomography and cognitive function in first diagnosed Parkinson’s disease patients

ObjectiveEvaluation of the correlation between 18F-fluorodeoxyglucose-positron emission tomography (18F-FDG PET) and cognitive function in first-diagnosed and untreated Parkinson’s disease (PD) patients.Materials and methodThis cross-sectional study included 84 first diagnosed and untreated PD patients. The individuals were diagnosed by movement disorder experts based on the 2015 MDS Parkinson’s disease diagnostic criteria. The patients also underwent 18F-FDG PET scans and clinical feature assessments including the Montreal Cognitive Assessment (MoCA) scale. Glucose metabolism rates were measured in 26 brain regions using region of interest (ROI) and pixel-wise analyses with displayed Z scores. The cognitive function was assessed by professionals using the MoCA scale, which covers five cognitive domains. Spearman’s linear correlation and linear regression models were used to compare the correlations between 18F-FDG metabolism in each brain region and cognitive domain, using SPSS 25.0 software.ResultThe results indicated a positive correlation between executive function and glucose metabolism in the lateral prefrontal cortex of the left hemisphere (p = 0.041). Additionally, a positive correlation between memory function and glucose metabolism in the right precuneus (p = 0.014), right lateral occipital cortex (p = 0.017), left lateral occipital cortex (p = 0.031), left primary visual cortex (p = 0.008), and right medial temporal cortex (p = 0.046). Further regression analysis showed that for every one-point decrease in the memory score, the glucose metabolism in the right precuneus would decrease by 0.3 (B = 0.30, p = 0.005), the glucose metabolism in the left primary visual cortex would decrease by 0.25 (B = 0.25, p = 0.040), the glucose metabolism in the right lateral occipital cortex would decrease by 0.38 (B = 0.38, p = 0.012), and the glucose metabolism in the left lateral occipital cortex would decrease by 0.32 (B = 0.32, p = 0.045).ConclusionThis study indicated that cognitive impairment in PD patients mainly manifests as changes in executive function, visual-spatial function and memory functions, while glucose metabolism mainly decreases in the frontal and posterior cortex. Further analysis shows that executive function is related to glucose metabolism in the left lateral prefrontal cortex. On the other hand, memory ability involves changes in glucose metabolism in a more extensive brain region. This suggests that cognitive function assessment can indirectly reflect the level of glucose metabolism in the relevant brain regions

Characteristics and influencing factors of 11C-CFT PET imaging in patients with early and late onset Parkinson’s disease

ObjectiveThis study aims to explore the difference between 11C-methyl-N-2β-carbomethoxy-3β-(4-fluorophenyl)-tropanel (11C-CFT) positron emission tomography (PET) imaging in the early-onset Parkinson’s disease (EOPD) and late-onset Parkinson’s disease (LOPD), and to analyze the correlation between 11C-CFT PET imaging and disease duration, Hoehn &amp; Yahr (H&amp;Y) stage, motor symptoms, and non-motor symptoms in patients with idiopathic Parkinson’s disease (PD), so as to explore its application value in assessing the severity of Parkinson’s disease.Materials and methodsA total of 113 patients with idiopathic PD were included in this study. The patients were divided into EOPD and LOPD groups according to the age of 60 years, of which 58 were early-onset and 55 were late-onset. All patients underwent 11C-CFT PET imaging and manually sketched regions of interest (ROI) to delineate the caudate nucleus, anterior putamen, and posterior putamen ROI layer-by-layer, and the corresponding values were recorded. Clinical data [age of onset, disease duration, H&amp;Y stage, total Unified Parkinson’s Disease Rating Scale (UPDRS) score, UPDRS III score, tremor score, postural instability/gait difficulty (PIGD) score, rigidity score, bradykinesia score, and Montreal Cognitive Assessment (MoCA) score] were collected from all patients. The differences in striatal 11C-CFT uptake between patients with EOPD and LOPD were compared, and the correlation between striatal 11C-CFT uptake and the clinical data of patients with idiopathic PD was evaluated.ResultsThe caudate nucleus 11C-CFT uptake was higher in EOPD than in the LOPD group (t = 3.002, p = 0.003). 11C-CFT uptake in the caudate nucleus in patients with PD was negatively correlated with the age of onset, H&amp;Y stage, disease duration, total UPDRS score, UPDRS III score, rigidity score, and bradykinesia score (p &lt; 0.05). The anterior and posterior putamen 11C-CFT uptake was negatively correlated with H&amp;Y stage, disease duration, total UPDRS score, UPDRS III score, PIGD score, rigidity score, and bradykinesia score (p &lt; 0.05).Conclusion11C-CFT PET provides an objective molecular imaging basis for the difference in disease progression rates between patients with EOPD and LOPD. Secondly, 11C-CFT PET can be used as an important objective indicator to assess disease severity and monitor disease progression

Transient low T3 syndrome in patients with COVID-19: a new window for prediction of disease severity

ObjectiveTo investigate the relationship of low T3 syndrome with disease severity in patients with COVID-19.MethodsThe clinical data of 145 patients with COVID-19 were retrospectively collected, and patients were divided into a low T3 group and a normal T3 group. Logistic regression models were used to assess predictive performance of FT3. Receiver operating characteristic (ROC) analysis was used to evaluate the use of low T3 syndrome in predicting critical disease. Kaplan-Meier analysis was used to analyze the impact of low T3 syndrome on mortality.ResultsThe prevalence of low T3 level among COVID-19 patients was 34.48%. The low T3 group was older, and had lower levels of hemoglobin, lymphocytes, prealbumin, and albumin, but higher levels of white blood cells, neutrophils, CRP, ESR, and D-dimer (all p&lt;0.05). The low T3 group had greater prevalences of critical disease and mortality (all p &lt;0.05). Multivariate logistic regression analysis showed that the Lymphocytes, free T3 (FT3), and D-dimer were independent risk factors for disease severity in patients with COVID-19. ROC analysis showed that FT3, lymphocyte count, and D-dimer, and all three parameters together provided reliable predictions of critical disease. Kaplan-Meier analysis showed the low T3 group had increased mortality (p&lt;0.001). Six patients in the low T3 group and one patient in the normal T3 group died. All 42 patients whose T3 levels were measured after recovery had normal levels after discharge.ConclusionPatients with COVID-19 may have transient low T3 syndrome at admission, and this may be useful for predicting critical illness