OR-016 Inflammatory responses associated with cortisol and CK after intensive endurance exercise

Objective The aim of this study was  to investigate the effect of acute intensive endurance exercise on circulating leucocyte and cytokine levels in trained kayakers, and to explore inflammatory responses associated with stress hormone and muscle damage. Methods Nine male trained kayakers (average age 21.00±3.94 years, average training history 5.56±1.88 years, average height 187.70±4.69 cm, average weight 89.25±8.66 kg, average body fat% 10.96±4.15%, and average VO2max 4.33±0.62 l/min ) participated in this study. All participants were well informed with the procedures and gave written informed consent. All participats completed a 3000m runing test 2 h after a normal breakfast. Participants were rested for 24 h prior to the exercise test, and kept fasting in 90 min after running test, but drunk water ad arbitrium. Mean heart rate was recorded by a Polar heart rate meter, the earlobe capillary blood lactate was tested with a portable lactate analyzer 3 min after running. Venous blood samples were taken from antecubital vein before (T0) , 15 min (T1) and 90 min (T2) after exercise. Whole blood samples were used for leucocyte counting and its subpopulation counting immediately by Coulter Counter， CD3+, CD4+, CD8+, CD4+ /CD8+, natural killer cells (NK) was evaluated by flow cytometry (Beckman Coulter), T cells were defined as CD3+ lymphocytes and NK was defined as CD3-CD16+CD56+ lymphocyte. Plasma and serum were isolated at 4oC. Plasma ACTH and serum testosterone, cortisol were assayed by CLIA (Immulite 2000), Serum IL-4, IL-6 were detected by ELISA (ST-360). Serum creatine kinase (CK) was assayed by autoanalyzer (Beckman CoulterAU680). Post-exercise concentrition of parameters in plasma and serum was corrected according to the formula by Dill. The data was analysed by IBM SPSS Statistics for Windows, Version 21.0. Continuous variables with normal distribution were presented as mean±standard deviation. Differences between the 3 time points were tested using repeated measures analysis of variance. To classicfy immune−endocrine relationships and  immune−muscle damage relationships, changes of inflammatory parameters were analysed in relation to testosterone, cortisol, ACTH and CK using Pearson's correlation coefficient respectively. Correlation between changes of testosterone, cortisol, ACTH and inflammatory parameters was tested to verify stress hormone related inflammation reponse, correlation between changes of CK and inflammatory parameters was tested to evaluate muscle damage related inflammation response. Only changes which from T1 or T2 differ significantly from T0, and from T1 differ significantly from T2 were included in the correlation analysis. The level of significance was set at p≤0.05. Results Paticipants finished 3000m running with an average time of 715 ± 33.0 Sec, and the average heart rate was 167.00±12.88 b/min, Blood lactate concentration was 12.31±1.91 mmol/l. The result showed significant increases for serum cortisol, serum CK, NK, neutrophile granulocyte% ( GR%), and significant deccreases for testosterone, CD4+, CD4+/CD8+, lymphocyte% (LY%) immedialtly after intensive running. Change of leucocyte count was not significant as a result of inceased NK, GR and decreased CD4+, LY. Compared with result of T1, significant increases of CD4+/CD8+, CD4+, leucocyte count, GR%, and significant decreases of ACTH, cortisol, CD8+, CD3+, LY% were observed at T2, leucocyte count increased with a trend for GR. Furthermore, compared the results of T2 with that of T0, participants presented increased CD4+/CD8+, CK, leucocyte, GR% and decreased testoterone, cortisol, ACTH, CD8+, CD3+, LY%, monocyte% (MO%) at T2. Compared all the levels of IL-4 and IL-6 at T0,T1 and T2 with each other, differences between any two were not significant. The time course of the changes revealed that significantly increased cortisol, NK and decreased CD4+ from T0 to T1, which reovered at T2 compared with T0 were fast response and fast recovery (FR-FR) parameters; Significantly increased CK, CD4+/CD8+, GR%, MO% and decreased testosterone,  LY% from T0 to T1, which didn’t reovered at T2 compared with T0 were fast response and slow recovery (FR-SR) parameters; Significantly increased leucocyte from T0 to T2 and decreased ACTH, CD3+, CD8+ from T1 to T2 were slow response and slow recovery (SR-SR) parameters; IL-4 and IL-6 were nonsensitive response (NR) parameters. Signifcant correlation was observed between ΔcortisolT1-T0 and ΔNKT1-T0 (r=0.78, p=0.04), ΔcortisolT2-T1 and ΔleucocyteT2-T1  (r=-0.70, p=0.04),  ΔACTHT2-T1 and ΔleucocyteT2-T1 (r=-0.76, p=0.02), ΔACTHT2-T0 and ΔCD8+T2-T0 (r=-0.79, p=0.03). No changes (ΔT1-T0, ΔT2-T1, ΔT2-T0) of inflammatory parameters correlated significantly to changes of testosterone and CK. But the correlation analysis revealed a signifcant correlation between ΔCKpost-exercise and Δcortisolpost-exercise (ΔT2-T1, r=0.90, p=0.001; ΔT2-T0, r=0.78, p=0.01). Conclusions Findings demonstrate different time-course responses and recovery of inflammatory parameters to intensive endurance exercise,athletes and coaches should consider these　different recovery time-courses in the subsenquent training session after intensive endurance training. Correlation analysis between cortisol, CK and inflammatory parameters indicates that inflammatory response is a stress hormone other than a muscle damage tuned process, and on the contrary, the correlation between post-exercise changes of CK and cortisol suggests a hint that inflammatory response caused by cortisol may contribute to the post-exercise muscle damage but not that during exercising. Correlation analysis and time-course analysis reveal that cortisol exerting an acute effect on NK, but maybe a false time-lag effect on leucocyte. This research investigated the inflammatory reponses to intensive endurance exercise by correlation analysis, but inflammary reponse is a multifactor process, and the conclusion is still challenging, further research is necessary to understand the underlying mechanism

Subduction Dynamics at the Northwestern Pacific Slab Edge: Constraints of Tomography in Kamchatka

Kamchatka is located at the northwestern edge of the Pacific plate. The Pacific slab is subducting into the mantle along the Kamchatka trench. Are there any differences in subduction features near the edge? To answer this question, seismic tomography was applied to 77,141 P-wave arrival times of 2239 local earthquakes and 75 teleseismic events recorded at 76 permanent stations to study the three-dimensional velocity structure to a depth of 700 km below Kamchatka. A clear high-velocity anomaly was evident beneath the study region, which is consistent with the distribution of intermediate-depth and deep-focus earthquakes. This high-velocity anomaly was interpreted as the subducting Pacific slab with its subduction angle and depth gradually increasing from the north to south along the Kamchatka trench. Another high-velocity anomaly appeared in the mantle transition zone and the uppermost lower mantle, which may reflect a piece of detached oceanic lithosphere due to melting of the subducting slab near the slab edge. Two slab windows were also found, through which hot mantle materials flowed from the subslab to the upper-mantle wedge. Large-scale low-velocity anomalies exist under the volcanic front, which reflect hot and wet upwelling flow in the mantle wedge due to the slab dehydration

Using Successive Self-Nucleation and Annealing to Detect the Solid−Solid Transitions in Poly(hexamethylene carbonate) and Poly(octamethylene carbonate)

Unformatted post-print version of the accepted articleSolid-solid transitions in poly (hexamethylene carbonate) (PC6) and poly (octamethylene carbonate) (PC8), denoted δ to α transition, have been investigated, using self-nucleation and Successive Self-nucleation and Annealing (SSA) technique. The SSA protocol was performed in-situ for thermal (differential scanning calorimetry (DSC)), structural (Wide-angle X-ray Scattering (WAXS)), and conformational (Fourier-transformed Infrared Spectroscopy (FT-IR)) characterization. The final heating after SSA fractionation displayed an enhanced (compared to a standard second DSC heating scan) endothermic and unfractionated peak signal at low temperatures corresponding to the δ to α transition. The improved (i.e., higher enthalpy and temperature than in other crystallization conditions) δ to α transition signal is produced by annealing the thickest lamellae made up by α and β phase crystals after SSA treatment. As thicker lamellae are annealed, more significant changes are produced in the δ to α transition, demonstrating the transition dependence on crystal stability, thus, on the crystallization conditions. The ability of SSA to significantly enhance the observed solid-solid transitions makes it an ideal tool to detect and study this type of transitions. In-situ WAXS reveals that the δ to α transition corresponds to a change in the unit cell dimensions, evidenced by an increase in the d-spacing. This implies a more efficient chain packing in the crystal, for both samples, in the δ phase (lower d-spacing at low temperatures) than in the α phase (higher d-spacing at high temperatures). The chain packing differences are explained through in-situ FT-IR measurements that show the transition from ordered (δ phase) to disordered (α phase) methylene chain conformations.We would like to acknowledge financial support provided by the National Key R&D Program of China (2017YFE0117800) and the National Natural Science Foundation of China (51820105005, 21922308, and 52050410327). We also acknowledge financial support from the BIODEST project; this project has received funding from the European Union’s Horizon 2020 research and innovation program under the Marie Skłodowska-Curie Grant Agreement No. 778092. This work has also received funding from MINECO through project MAT2017-83014-C2-1-P and from the Basque Government through grant IT1309-19. R.A.P.-C is supported by the China Postdoctoral Science Foundation (2020M670462). G.L. is grateful to the Youth Innovation Promotion Association of the Chinese Academy of Sciences (Y201908). We also thank the BSRF (beamline 1W2A) for providing beamtime

Solid–Solid Crystal Transitions (δ to α) in Poly(hexamethylene carbonate) and Poly(octamethylene carbonate)

Unformatted post-print version of the accepted articlePoly (hexamethylene carbonate) (PC6) and poly (octamethylene carbonate) (PC8) were studied under different crystallization conditions. Using differential scanning calorimetry (DSC), a new solid-solid transition, denoted α to δ transition, was detected at low temperatures (<RT) in both PC6 and PC8 samples. The α to δ transition was represented by exothermic (i.e., α to δ) (−6 °C (PC6) and −20 °C (PC8)) and endothermic peaks (i.e., δ to α) (15 °C (PC6) and 28 °C (PC8)), during cooling and heating DSC scans, respectively. Isothermal tests revealed that this solid-solid transition depends on the specific thermal history, since it is not observed at isothermal temperatures higher than room temperature. Still, it is detected in the subsequent cooling and heating scans. Wide-angle X-ray scattering (WAXS) and Fourier-transform infrared spectroscopy (FT-IR) experiments were performed at identical conditions to those by DSC. WAXS experiments showed lower d-spacings in the δ phase than in the α one, corresponding to a unit cell shrinkage, explained by a more efficient packing of the methylene groups in the δ phase. The δ phase is also characterized, according to FT-IR experiments, by more ordered conformation of the methylene groups (i.e., reflected in the appearance of a new absorption band) compared to the less ordered conformation in the α phase.This work is supported by the National Key R&D Program of China (2017YFE0117800) and the National Natural Science Foundation of China (51820105005, 21922308, and 52050410327). We would like to thank the financial support provided by the BIODEST project; this project has received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement no. 778092. This work has also received funding from MINECO through project MAT2017-83014-C2-1-P and from Basque Government through grant IT1309-19. R.A.P.-C is supported by the China Postdoctoral Science Foundation (2020M670462). G.L. is grateful to the Youth Innovation Promotion Association of the Chinese Academy of Sciences (Y201908). We also thanks to the BSRF (beamline 1W2A)

Study on stability and homology analysis of arginine kinase from oyster

Objective To identify the natural protein arginine kinase (AK) extracted from oysters, and to understand its basic properties and homology. Methods AK was isolated and purified from oysters by ammonium sulfate salting out and anion exchange, and the relative molecular mass and secondary structure were determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and circular dichroism (CD) spectroscopy. Its stability was also studied. The amino acid sequences of oyster AK and 11 other crustacean and mollusk allergens were compared by bioinformatics software, and their homology was analyzed. Results The natural protein with a relative molecular weight of 40 kDa was oyster AK. AK was neither heat-resistant nor acid-resistant. Oyster AK has high homology of mollusk allergen AK amino acid sequence, and homology of crustacean amino acid sequence is 55%-60%. Conclusion Natural AK is extracted from oysters, and the stability and homology are basically understood. It will lay the foundation for comprehensive research on sensitization and sensitization mechanism of oysters

Genome Editing of Pik3cd Impedes Abnormal Retinal Angiogenesis

Abnormal angiogenesis is associated with myriad human diseases including proliferative diabetic retinopathy. Signaling transduction via phosphoinositide 3-kinases (PI3Ks) plays a critical role in angiogenesis. Herein, we showed that p110δ, the catalytic subunit of PI3Kδ, was highly expressed in pathological retinal vascular endothelial cells (ECs) in a mouse model of oxygen-induced retinopathy (OIR) and in fibrovascular membranes from patients with proliferative diabetic retinopathy. To explore novel intervention with PI3Kδ expression, we developed a recombinant dual adeno-associated viral (rAAV) system for delivering CRISPR/Cas9 in which Streptococcus pyogenes (Sp) Cas9 expression was driven by an endothelial specific promoter of intercellular adhesion molecule 2 (pICAM2) to edit genomic Pik3cd, the gene encoding p110δ. We then demonstrated that infection of cultured mouse vascular endothelial cells with the dual rAAV1s of rAAV1-pICAM2-SpCas9 and rAAV1-SpGuide targeting genomic Pik3cd resulted in 80% DNA insertion/deletion in the locus of genomic Pik3cd and 70% depletion of p110δ expression. Furthermore, we showed that in the mouse model of OIR editing retinal Pik3cd with the dual rAAV1s resulted in not only a significant decrease in p110δ expression, and Akt activation, but also a dramatic reduction in pathological retinal angiogenesis. These findings reveal that Pik3cd editing is a novel approach to treating abnormal retinal angiogenesis

Correction to: Genome Editing of Pik3cd Impedes Abnormal Retinal Angiogenesis, by Wu et al. Hum Gene Ther 2023;34(1-2):30-41; doi: 10.1089/hum.2022.079

In the January 2023 issue of Human Gene Therapy (vol. 34, no. 1-2; 30–41), the article titled Genome Editing of Pik3cd Impedes Abnormal Retinal Angiogenesis, by Wu et al. requires correction. The author byline originally appeared with the 13th author's name incorrectly published as GuomingZhao Wenyi Wu,1,2,3 Gaoen Ma,4 Hui Qi,5 Lijun Dong,5 Fang Chen,6 Yun Wang,5 Xingxing Mao,5 Xiaoqing Guo,2,3 Jing Cui,7 Joanne Aiko Matsubara,7 Bart Vanhaesebroeck,8 Xiaohe Yan,5Guoming Zhao,5 Shaochong Zhang,5,* and Hetian Lei 5,* The correct spelling of the author's name is GuomingZhang The online version of the article has been corrected to reflect this. The authors apologize for the error

Non-invasive Amide Proton Transfer Imaging and ZOOM Diffusion-Weighted Imaging in Differentiating Benign and Malignant Thyroid Micronodules

Background: Pre-operative non-invasive differentiation of benign and malignant thyroid nodules is difficult for doctors. This study aims to determine whether amide proton transfer (APT) imaging and zonally oblique multi-slice (ZOOM) diffusion-weighted imaging (DWI) can provide increased accuracy in differentiating benign and malignant thyroid nodules.Methods: This retrospective study was approved by the institutional review board and included 60 thyroid nodules in 50 patients. All of the nodules were classified as malignant (n = 21) or benign (n = 39) based on pathology. It was meaningful to analyze the APT and apparent diffusion coefficient (ADC) values of the two groups by independent t-test to identify the benign and malignant thyroid nodules. The relationship between APT and ZOOM DWI was explored through Pearson correlation analysis. The diagnostic efficacy of APT and ZOOM DWI in determining if thyroid nodules were benign or malignant was compared using receiver operating characteristic (ROC) curve analysis.Results: The mean APTw value of the benign nodules was 2.99 ± 0.79, while that of the malignant nodules was 2.14 ± 0.73. Additionally, there was a significant difference in the APTw values of the two groups (P &lt; 0.05). The mean ADC value of the benign nodules was 1.84 ± 0.41, and was significantly different from that of the malignant nodules, which was 1.21 ± 0.19 (P &lt; 0.05). Scatter point and Pearson test showed a moderate positive correlation between the APT and ADC values (P &lt; 0.05). The ROC curve showed that the area under the curve (AUC) value of ZOOM DWI (AUC = 0.937) was greater than that of APT (AUC = 0.783) (P = 0.028).Conclusion: APT and ZOOM DWI imaging improved the accuracy of distinguishing between benign and malignant thyroid nodules. ZOOM DWI is superior to APTw imaging (Z = 2.198, P &lt; 0.05)

Real-time Monitoring for the Next Core-Collapse Supernova in JUNO

Core-collapse supernova (CCSN) is one of the most energetic astrophysical events in the Universe. The early and prompt detection of neutrinos before (pre-SN) and during the SN burst is a unique opportunity to realize the multi-messenger observation of the CCSN events. In this work, we describe the monitoring concept and present the sensitivity of the system to the pre-SN and SN neutrinos at the Jiangmen Underground Neutrino Observatory (JUNO), which is a 20 kton liquid scintillator detector under construction in South China. The real-time monitoring system is designed with both the prompt monitors on the electronic board and online monitors at the data acquisition stage, in order to ensure both the alert speed and alert coverage of progenitor stars. By assuming a false alert rate of 1 per year, this monitoring system can be sensitive to the pre-SN neutrinos up to the distance of about 1.6 (0.9) kpc and SN neutrinos up to about 370 (360) kpc for a progenitor mass of 30$M_{\odot}$ for the case of normal (inverted) mass ordering. The pointing ability of the CCSN is evaluated by using the accumulated event anisotropy of the inverse beta decay interactions from pre-SN or SN neutrinos, which, along with the early alert, can play important roles for the followup multi-messenger observations of the next Galactic or nearby extragalactic CCSN.Comment: 24 pages, 9 figure