A20 (TNFAIP3) alleviates viral myocarditis through ADAR1/miR-1a-3p-dependent regulation

Objective To investigate the effect of A20 and how A20 is regulated in viral myocarditis (VMC). Methods BABL/C mice, primary neonatal rat cardiomyocytes and H9c2 cells were infected with Coxsackie virus B3 (CVB3) to establish animal and cellular models of VMC. H&E staining revealed the pathologic condition of myocardium. ELISA measured the serum levels of creatine kinase, creatine kinase isoenzyme and cardiac troponin I. The effects of A20, miR-1a-3p and ADAR1 were investigated using gain and loss of function approaches. ELISA measured the levels of IL-6, IL-18 and TNF-α in serum or cell culture supernatant. TUNEL staining and flow cytometry assessed the apoptosis of myocardium and cardiomyocytes, respectively. RNA-binding protein immunoprecipitation and dual-luciferase reporter assays verified the binding between A20 and miR-1a-3p. Co-immunoprecipitation assay verified the binding between ADAR1 and Dicer. Results A20 was underexpressed and miR-1a-3p was overexpressed in the myocardium of VMC mice as well as in CVB3-infected cardiomyocytes. Overexpression of A20 suppressed cardiomyocyte inflammation and apoptosis in vivo and in vitro. miR-1a-3p promoted CVB3-induced inflammation and apoptosis in cardiomyocytes by binding to A20. The expression of miR-1a-3p was regulated by ADAR1. ADAR1 promoted the slicing of miR-1a-3p precursor by binding to Dicer. Conclusion A20, regulated by ADAR1/miR-1a-3p, suppresses inflammation and cardiomyocyte apoptosis in VMC.

Biochemical profile in mixed martial arts athletes

The study aimed to evaluate changes in selected biochemical indicators among mixed martial arts competitors in subsequent periods of the training cycle. The research involved 12 mixed martial arts athletes aged 25.8 ± 4.2 years competing in the intermediate category. Selected somatic indicators were measured twice. Biochemical indicators were assessed five times during the 14-week study period. Serum concentrations of testosterone, cortisol, uric acid, myoglobin, total protein, interleukin 6, and tumor necrosis factor, as well as creatine kinase activity were determined. One hour after sparring completion, there were significant increases in cortisol (by 54.9%), uric acid (22.0%), myoglobin (565.0%), and interleukin 6 (280.3%) as compared with the values before the simulated fight. The highest creatine kinase activity (893.83 ± 139.31 U/l), as well as tumor necrosis factor (3.93 ± 0.71 pg/ml) and testosterone (5.83 ± 0.81 ng/ml) concentrations (p = 0.00) were recorded 24 hours after the simulation. Systematic observation of selected blood biochemical indicators in the training process periodization in mixed martial arts helps understand adaptive, compensatory, and regenerative mechanisms occurring in training athletes.

Individual-based Creatine Kinase Reference Values in Response to Soccer Match-play

The aim of the present study was to determine the creatine kinase reference limits for professional soccer players based on their own normal post-match response. The creatine kinase concentration was analyzed in response to official matches in 25 players throughout a 3-year period. Samples were obtained between 36–43 hours following 70 professional soccer matches and corresponded to 19.1±12.1 [range: 6–49] samples per player. Absolute reference limits were calculated as 2.5th and 97.5th percentile of the samples collected. Creatine kinase values were also represented as a percentage change from the individual’s season mean and represented by 90th, 95th and 97.5th percentiles. The absolute reference limits for creatine kinase concentration calculated as 97.5th and 2.5th percentiles were 1480 U.L−1 and 115.8 U.L−1, respectively. The percentage change from the individual’s season mean was 97.45±35.92% and players were in the 90th, 95th and 97.5th percentiles when the percentages of these differences were 50.01, 66.7, and 71.34% higher than player’s season mean response, respectively. The data allowed us to determine whether the creatine kinase response is typical or if it is indicative of a higher than normal creatine kinase elevation and could be used as a practical guide for detection of muscle overload, following professional soccer match-play.

Prevotellaceae produces butyrate to alleviate PD-1/PD-L1 inhibitor-related cardiotoxicity via PPARα-CYP4X1 axis in colonic macrophages

Background Immune checkpoint inhibitor-related cardiotoxicity is one of the most lethal adverse effects, and thus, the identification of underlying mechanisms for developing strategies to overcome it has clinical importance. This study aimed to investigate whether microbiota-host interactions contribute to PD-1/PD-L1 inhibitor-related cardiotoxicity. Methods A mouse model of immune checkpoint inhibitor-related cardiotoxicity was constructed by PD-1/PD-L1 inhibitor BMS-1 (5 and 10 mg/kg), and cardiomyocyte apoptosis and cardiotoxicity were determined by hematoxylin and eosin, Masson’s trichome and TUNEL assays. 16S rRNA sequencing was used to define the gut microbiota composition. Gut microbiota metabolites short-chain fatty acids (SCFAs) were determined by HPLC. The serum levels of myocardial enzymes (creatine kinase, aspartate transaminase, creatine kinase-MB and lactate dehydrogenase) and the production of M1 factors (TNF-α and IL-1β) were measured by ELISA. The colonic macrophage phenotype was measured by mmunofluorescence and qPCR. The expression of Claudin-1, Occludin, ZO-1 and p-p65 was measured by western blot. The gene expression of peroxisome proliferator-activated receptor α (PPARα) and cytochrome P450 (CYP) 4X1 was determined using qPCR. Statistical analyses were performed using Student’s t-test for two-group comparisons, and one-way ANOVA followed by Student–Newman–Keul test for multiple-group comparisons. Results We observed intestinal barrier injury and gut microbiota dysbiosis characterized by Prevotellaceae and Rikenellaceae genus depletion and Escherichia-Shigella and Ruminococcaceae genus enrichment, accompanied by low butyrate production and M1-like polarization of colonic macrophages in BMS-1 (5 and 10 mg/kg)-induced cardiotoxicity. Fecal microbiota transplantation mirrored the effect of BMS-1 on cardiomyocyte apoptosis and cardiotoxicity, while macrophage depletion and neutralization of TNF-α and IL-1β greatly attenuated BMS-1-induced cardiotoxicity. Importantly, Prevotella loescheii recolonization and butyrate supplementation alleviated PD-1/PD-L1 inhibitor-related cardiotoxicity. Mechanistically, gut microbiota dysbiosis promoted M1-like polarization of colonic macrophages and the production of proinflammatory factors TNF-α and IL-1β through downregulation of PPARα-CYP4X1 axis. Conclusions Intestinal barrier dysfunction amplifies PD-1/PD-L1 inhibitor-related cardiotoxicity by upregulating proinflammatory factors TNF-α and IL-1β in colonic macrophages via downregulation of butyrate-PPARα-CYP4X1 axis. Thus, targeting gut microbiota to polarize colonic macrophages away from the M1-like phenotype could provide a potential therapeutic strategy for PD-1/PD-L1 inhibitor-related cardiotoxicity. Graphical abstract

Plasma and tissue enzyme activities of banded water snakes (Nerodia fasciata) and diamondback water snakes (Nerodia rhombifer)

OBJECTIVE To measure plasma and tissue activities of alkaline phosphatase, alanine aminotransferase, aspartate aminotransferase (AST), creatine kinase, and γ-glutamyltransferase in 2 snake species. ANIMALS 6 banded water snakes (Nerodia fasciata) and 6 diamondback water snakes (Nerodia rhombifer). PROCEDURES Blood was collected via the ventral tail vein to measure plasma enzyme activities. Animals were then euthanized, and samples of 9 tissues were collected from each snake: skeletal muscle, cardiac muscle, liver, spleen, lung, kidney, testicle, pancreas, and gallbladder. Tissues were frozen for 30 days, then homogenized and processed. Supernatants were collected and analyzed within 24 hours of processing. A linear mixed model was used to determine differences in enzyme activity between tissues and species and assess interactions between tissues and species. RESULTS Activities of all enzymes were found to differ significantly among tissues. There were also significant differences between species for all enzyme activities, except AST activity. The kidney had the highest alanine aminotransferase and γ-glutamyltransferase activities. Alkaline phosphatase activity was significantly highest in liver and kidney tissues than in other tissue. Creatine kinase activity was highest in skeletal muscle, followed by cardiac muscle and kidney. AST activity was present in all tissues evaluated, but was highest in liver, kidney, and cardiac muscle in both species. CLINICAL RELEVANCE Results reinforced the importance of characterizing the origin of tissue enzymes in reptiles to improve our understanding of biochemistry results and highlighted the differences that can exist in tissue enzyme activities between closely related species.

Perbandingan Oedema Paru Akut Yang Disebabkan Oleh Tingkat Kardiogenik dan Non Kardiogenik Dengan Mortalitas Dan Morbiditas Pasien

Latar Belakang: Prognosis jangka panjang edema paru akut (APE) tetap tidak jelas. Metode dan Hasil: Kami mengevaluasi data demografis, ekokardiografi, dan angiographic dari 806 pasien berturut-turut dengan APE dengan (CAD) dan tanpa penyakit arteri koroner (non-CAD) yang diterima dari tahun 2000 hingga 2010. Perbedaan antara rumah sakit dan kematian jangka panjang dan prediktornya juga dinilai. Pasien CAD (n = 638) lebih tua dan memiliki insiden diabetes dan penyakit vaskular perifer yang lebih tinggi daripada non-CAD (n = 168), dan fraksi ejeksi yang lebih rendah. Kematian di rumah sakit serupa pada kedua kelompok (26,5% vs 31,5%; P = 0,169) tetapi kekambuhan KERA lebih tinggi pada pasien CAD (17,3% vs 6,5%; P<0.001). Usia, masuk tekanan darah sistolik, kekambuhan APE, dan kebutuhan inotropics atau intubasi endotrakcheal adalah prediktor independen utama kematian di rumah sakit. Sebaliknya, kematian secara keseluruhan (70,0% vs 57,1%; P = 0,002) dan penerimaan kembali untuk gagal jantung nonfatal setelah tindak lanjut 45 bulan (10-140; 17,3% vs 7,6%; P = 0,009) lebih tinggi pada CAD daripada pasien non-CAD. Usia, penyakit vaskular perifer, dan puncak creatine kinase MB selama rawat inap indeks, tetapi bukan fraksi ejeksi, adalah prediktor independen utama dari kematian secara keseluruhan, sedangkan revaskularisasi koroner atau operasi valvular bersifat protektif. Intervensi ini sebagian besar dilakukan selama indeks rawat inap (294 dari 307; 96%) dan tidak pasien yang diintervensi menunjukkan profil risiko yang lebih tinggi. Kesimpulan: Kematian jangka panjang di APE tinggi dan lebih tinggi pada CAD daripada pada pasien non-CAD. Mengingat prediktor kematian di rumah sakit dan jangka panjang yang berbeda di sini dijelaskan, yang tidak selalu melibatkan fungsi sistolik, dapat dibayangkan bahwa program intervensi yang lebih agresif dapat meningkatkan kelangsungan hidup pada pasien berisiko tinggi.

CREATININE AND CREATINE KINASE RATIO IN BLOOD OF DIFFERENT BODY TYPES - A NEW APPROACH.

Purpose: The inconsistencies and variations of creatine kinase level due to modifiable and non-modifiable factors were the basis of this study. The aim was to find out the relationships between creatinine and creatine kinase in the blood of somatotypes. Methods: The 122 males, aged 10 to 20 years, were classified according to their somatotypes. Somatotypes were measured by the ISAK method. By standard laboratory methods, creatinine and creatine kinase estimate. The IBM SPSS version 24 is used for calculation. One way ANOVA followed by post hoc tests was performed to compare the variables among the three groups (p<0.05). Results: Creatinine level in the blood insignificantly deferred among the three somatotypes. The significant differences (p<0.05) were found in creatine kinase level in the blood and creatinine/creatine kinase ratio among the three dominant Somatotypes. Creatine Kinase was significantly higher in Ectomorphs (212 U/L) than Endomorphs. Ectomorphs and mesomorphs have crossed normal creatine kinase levels (35 -175 U/L). The creatinine/creatine kinase ratio was found highest in endomorphs and lowest in the ectomorphs and significantly differed in three Somatotypes. Conclusion: Creatinine production remains the same, indicating production of Creatinine is independent of specific body types. A significant higher Creatine Kinase level in Ectomorphs over Endomorphs showed fat content was not associated with it. Significant differences in Creatinine / Creatine Kinase ratio among Somatotypes suggested its relevance between cellular and morphological relationships and might uses as biomarkers.

Ascorbic Acid Significantly Decreases Creatine Kinase Plasma Levels in an Animal Model of Statin/Fibrate-Induced Myopathy

Background. Myopathy is one of the side effects of lipid-lowering drugs, especially statins and particularly when combined with a fibrate. To diagnose myopathy and determine its severity, the plasma levels of three enzymes, creatine kinase (CK), aldolase, and lactate dehydrogenase (LDH), are routinely measured. Physical exercise can aggravate the statin-associated muscular disease. The question is whether antioxidants like ascorbic acid (Vit. C) can prevent such myopathy. Methods. In this experiment, a combination of atorvastatin (ATV, 80 mg/kg/day) and gemfibrozil (GMF, 1000 mg/kg/day) orally for 10 days as well as exercise as forced swimming on days 8, 9, and 10 were used to induce myopathy. Ascorbic acid (50 mg/kg/day, orally) was added to ATV/GMF plus exercise regimen throughout the 10 days in the treatment group. Mean blood levels of CK, aldolase, and LDH were measured in addition to swimming tolerance times. Results. There was a significantly higher swimming tolerance time P < 0.05 and lower CK levels P < 0.01 in rats receiving ATV/GMF/Vit. C plus exercise compared with rats not taking Vit. C. LDH and aldolase did not decrease significantly. Conclusion. The results of this study showed that Vit. C can be effective in preventing myopathy caused by fat-lowering drugs.

Serum Creatinine and Creatine Kinase as an Indicator for Gingival Wound Healing in Rabbits by Supplementation of Pomegranate Seed Extract

Background: Pomegranate (Punica granatum) is an edible fruit that has been described as a medical and therapeutic functional food in the Middle East and the Mediterranean. Objective: To investigate the use of serum creatinine (Cr) level and creatine kinase (CK) activity as indicators for gingival wound healing process rate in rabbits, supplemented on pomegranate seed extract (PSE). Patients and Methods: A total of 45 rabbit males were used. They were divided into 3 groups; 5 rabbits as a baseline group that left without a buccal gingival wound. 20 rabbits (5 rabbits per each time interval) as a study group with buccal gingival wound with PSE supplementation, and another 20 rabbits (5 rabbits per each time interval) as a control group with gingival wound without PSE supplementation. A buccal gingival wound was created on the lower right central incisor, and the suture was removed after (7) days. Blood samples were collected for the baseline group and at time intervals; 3 hour, 1, 3, 7 days after creating the wound for both control and study groups to determine serum Cr and serum CK. Results: Serum Cr and CK significantly increased in all time intervals after gingival wounds, in the control group compared with baseline values. Whereas the levels significantly increased in rabbits receiving PSE at intervals of 3 hours, 1 and 3 days after gingival wound, they returned to the baseline values, seven days after gingival wound incision. Conclusion: Serum Cr and CK increase in the buccal gingival wound, while oral supplementation of PSE can decline them to the baseline value after a period of time, therefore these parameters can be used as indicators for gingival wound healing rate. Keywords: Punicagranatum seed extract, gingival wound healing, creatinine, creatine kinase.