METTL3, MTEEL14, and WTAP), “erasers” (FTO and ALKBH5), and “readers” (YTHDF2) in peripheral blood were determined by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The results demonstrated that the mRNA levels of METTL3, WTAP, FTO, ALKBH5, and YTHDF2 in peripheral blood from SLE patients were significantly decreased. The levels of ALKBH5 mRNA in SLE patients were associated with anti-dsDNA, antinucleosome, rash, and ulceration. Multivariate logistic regression analysis showed that the level of ALKBH5 mRNA in peripheral blood is a risk factor of SLE (P < 0.001). Moreover, our results suggested that there was a positive correlation between m6A“writers” (METTL3 and WTAP), “erasers” (FTO and ALKBH5), and “readers” (YTHDF2) in SLE patients. This study suggests that the mRNA level of ALKBH5 in peripheral blood may be involved in the pathogenesis of SLE.Although it has been proved that the epigenetic modification of DNA and histones is involved in the pathogenesis of systemic lupus erythematosus (SLE), there is no study to explore whether the modification of N6-methyladenosine (m6A) in RNA is involved. In this study, the mRNA levels of m6A “writers” ( The systemic lupus erythematosus (SLE) is a chronic and incurable autoimmune disease characterized by intermittent episodes of increased disease activity that require treatment with immunosuppressive agents , 2. AlthIn recent years, epigenetic modifications have been demonstrated to play an important role in the genesis and development of SLE , 9. N6-mMETTL3, MTEEL14, WTAP, FTO, ALKBH5, and YTHDF2 in peripheral blood were detected in SLE patients and analyzed for their correlation with clinical variables.Recent studies have demonstrated that m6A modification is associated with various human diseases , 16. HowA total of 51 patients that fulfilled the revised American College of Rheumatology criteria for SLE  were recThe peripheral blood collection and total RNA isolation were performed as described previously .METTL3, METTL14, WTAP, ALKBH5, FTO, and YTHDF2 transcripts were quantified with an ABI 7500 Real-time PCR System  by using SYBR® Premix Ex Taq™ II . The sequences of amplification primers for METTL3, METTL14, WTAP, ALKBH5, FTO, YTHDF2, and GAPDH are listed in ΔCt- method [Complementary DNA (cDNA) was obtained from purified total RNA samples by reverse transcription using a PrimeScript™ RT reagent kit . t method , 21.The items in blood routine and urinary routine were detected using Sysmex-x2100  and SysmexUF-1000i . Erythrocyte sedimentation rate (ESR) was determined by the BlessLBY-XC40B automatic ESR analyzer  according to the manufacturer's instructions. The levels of C-reactive protein (CRP), immunoglobulin G (IgG), complement 3 (C3), and complement 4 (C4) in serum were determined by nephelometry methods using IMMAGE800 system . Anti-dsDNA of IgG in serum was measured by commercially available enzyme-linked immunosorbent assay (ELISA) kits . Antiextractable nuclear antigen (Anti-ENA) was determined using line immunoassay kits  according to the manufacturer's instructions.t-test or the Mann-Whitney U-test was used to compare the data according to the normality. The Spearman method was used for correlation analysis. Logistic regression analysis was used for evaluating the risk factor. All the data were analyzed by GraphPad Prism version 5.0  and SPSS version 17.0 . P values < 0.05 were considered statistically significant.Student's METTL3, METTL14, WTAP, ALKBH5, FTO, and YTHDF2 in patients with SLE, the mRNA levels of these genes in peripheral blood were determined by qRT-PCR in SLE patients and HCs. Data showed that the mRNA levels of METTL3, WTAP, ALKBH5, and FTO in peripheral blood of SLE patients significantly decreased compared to HCs   (METTL14 was unchanged (P = 0.673) (To identify the expression levels of < 0.050) . The mRN= 0.097) . The mRN= 0.673) .METTL3, WTAP, ALKBH5, FTO, and YTHDF2 in peripheral blood in the screening stage, an independent cohort including 23 SLE patients and 12 HCs were recruited and detected for the mRNA levels of these genes in peripheral blood. The data from all subjects, including all 51 SLE patients and 38 HCs in screening stage and validation stage, demonstrated that the mRNA levels of METTL3, WTAP, ALKBH5, and FTO in peripheral blood were significantly lower in SLE patients than those in HCs    . Inconsi< 0.001) .METTL3, WTAP, ALKBH5, FTO, and YTHDF2 and autoantibodies were investigated in these SLE patients. As shown in ALKBH5 was significantly decreased in patients with positive anti-dsDNA compared to patients with negative anti-dsDNA. Furthermore, the level of peripheral blood ALKBH5 in SLE patients negatively correlated with the level of anti-dsDNA  (ALKBH5 was significantly decreased in patients with positive antinucleosome compared to patients with negative antinucleosome (P = 0.003) (ALKBH5 did not correlate with other anti-ENAs (data not shown). No obvious correlation was observed between the levels of peripheral blood METTL3, WTAP, FTO, and YTHDF2 and autoantibodies in SLE patients (data not shown).Production of multiple autoantibodies such as anti-double-stranded DNA (anti-dsDNA) and antiextractable nuclear antigens (Anti-ENAs) is one of the important characteristics of SLE. In 51 SLE patients, 44 SLE patients were tested for anti-dsDNA and 37 SLE patients were tested for anti-ENAs. The correlations between the mRNA levels of peripheral blood = 0.043) . And, th= 0.003) . The levMETTL3, WTAP, ALKBH5, FTO, and YTHDF2 in SLE patients were decreased and the level of ALKBH5 in peripheral blood was associated with autoantibody production. Considering that the production of autoantibodies plays an important role in the pathogenesis of SLE, the “enter method” of multivariate logistic regression was used to explore whether the expression of peripheral blood METTL3, WTAP, ALKBH5, FTO, and YTHDF2 was risk factors for SLE. As shown in METTL3, WTAP, ALKBH5, FTO, and YTHDF2 were obtained, Y = −326.128 X1(METTL3) − 74.188 (WTAP) − 414.645 X3(ALKBH5) − 15.042 X4(FTO) − 0.106 X2(YTHDF2) + 6.197. The results demonstrated that the decreased expression of ALKBH5 in peripheral blood was a risk factor for SLE (P < 0.001), while other molecules are not (P > 0.050). Furthermore, the mRNA level of peripheral blood ALKBH5 in other inflammatory disorders including RA, TB, and HBV-infected patients was detected and compared with that in SLE patients. Results showed that the mRNA level of peripheral blood ALKBH5 in SLE patients was significantly lower compared to RA patients (P = 0.003) (P < 0.0001) (P < 0.0001)  , HBV-inf 0.0001) , and TB  0.0001) .ALKBH5 in SLE patients was negatively correlated with the level of anti-dsDNA. Anti-dsDNA was proved to be involved in the genesis of SLE and can be served as markers for evaluating ongoing disease activity in SLE and early relapse of SLE. And, the level of anti-dsDNA in SLE patients was found to negatively correlate with the level of C3     did not correlate with SLEDAI, CRP, ESR, IgG, C3, C4, WBC, RBC, HGB, HCT, PLT, L, M, or N (data not shown). However, further analysis in the new-onset SLE patients showed that the mRNA level of peripheral blood METTL3 positively correlated with PLT     , the mRN= 0.028) , and the= 0.039) .Next, the relationship between the levels of these m6A modification-related molecules and drug therapy of SLE was analyzed. However, we found no significant difference between new-onset SLE patients and revisiting SLE patients (data not shown). Subsequently, a 15-day follow-up evaluation in 7 new-onset SLE patients who received regular treatment was performed, but there was still no difference (data not shown).METTL3 and WTAP in SLE patients with alopecia was significantly decreased than that in SLE patients without alopecia ; the mRNA level of peripheral blood ALKBH5 in SLE patients with rash and ulceration was significantly increased than that in SLE patients without rash and ulceration, respectively . No relationship was found between the levels of peripheral blood FTO, YTHDF2, and SLE symptoms         , ALKBH5 = 0.015) , FTO (rs< 0.001) , and YTH= 0.011) ; the mRN= 0.020) ; and the< 0.001)  and FTO = 0.015) .Epigenetic alterations, including DNA methylation and histone modifications, have been reported to contribute to SLE progression , 22, 23.METTL3, MTEEL14, WTAP, and ALKBH5 were significantly decreased in testicular germ cell tumors and the m6A modification in RNA of testicular germ cell tumors was controlled by METTL3, ALKBH5, and YTHDF2 [METTL3 and FTO during the osteogenic induction of BMSCs were increased [FTO, METTL3, METTL14, and WTAP were increased in patients with type-2 diabetes [METTL3 and WTAP), erasers (ALKBH5 and FTO), and reader (YTHDF2) were all decreased in peripheral blood from SLE patients. In accordance with the results of Yang et al. [METTL3, WTAP, ALKBH5, FTO, and YTHDF2 may play an important role in the pathogenesis of SLE.As the regulators of m6A methylation, METTL3, MTEEL14, WTAP, FTO, ALKBH5, and YTHDF2 play pivotal roles in the dynamic regulation of m6A modification. Nettersheim et al. showed that the mRNA levels of d YTHDF2 . Tian etncreased . In addidiabetes . These sg et al. , our resg et al. , our stuALKBH5 was significantly decreased in SLE patients compared to HCs, RA patients, TB patients, and HBV-infected patients. Furthermore, the level of peripheral blood ALKBH5 significantly decreased in SLE patients with positive anti-dsDNA or antinucleosome compared to SLE patients with negative anti-dsDNA or antinucleosome. And, the level of peripheral blood ALKBH5 in SLE patients was found to be negatively correlated with the level of anti-dsDNA. In addition, the level of peripheral blood ALKBH5 in SLE patients was associated with clinical symptoms of SLE such as rash, ulceration, and leukopenia. Logistic regression analysis revealed that decreased mRNA level of ALKBH5 in peripheral blood was a risk factor for SLE.SLE is a systemic autoimmune disease characterized by elevated autoimmune antibodies, such as anti-dsDNA and antinucleosome. It is well known that anti-dsDNA and antinucleosome are pathogenic autoantibodies involved in organ damage of SLE –34. In tALKBH5 may be involved in the pathogenesis of SLE and may also serve as a potential biomarker for SLE diagnosis and severity evaluation.Considering that anti-dsDNA and antinucleosome are involved in the pathogenesis of SLE and are specific diagnostic biomarkers of SLE, our results suggested that the expression of peripheral blood METTL3, METTL14, WTAP, ALKBH5, FTO, and YTHDF2 in SLE patients. In this study, we found that the mRNA levels of METTL3, WTAP, ALKBH5, FTO, and YTHDF2 in peripheral blood from SLE patients were significantly decreased. Additionally, this study established the correlations between the mRNA level of ALKBH5 and the production of autoantibodies, clinical features, which might improve our understanding of the role of ALKBH5 in SLE.In conclusion, it is the first study to our knowledge that explored the mRNA levels of peripheral blood


Background: Chronic kidney disease (CKD) constitutes a public health issue that is estimated to affect more than 10% of global population. Over 100 million people have CKD in China. For the first time, this cross-sectional study was designed to determine whether multiple measures of mineral metabolism had a significant association with renal function in Chinese centenarians.Methods: China Hainan Centenarian Cohort Study was conducted in 18 cities and counties of Hainan Province, China. It utilized the standard methodology for home interview and blood analyses in 750 centenarians including 608 females and 142 males.Results: All centenarians have a median (range) age of 102 (100–115) years, and median (range) level of estimated glomerular filtration rate (eGFR) was 63 (16–138) ml/min/1.73 m2. There were 318 centenarians (42.4%) with eGFR levels <60 ml/min/1.73 m2. In simple correlation and multivariate linear regression analyses, serum phosphorus, osteocalcin, β-crosslaps, total procollagen type 1 N-terminal propeptide (TP1NP), and parathyroid hormone (PTH) levels were negatively associated with eGFR levels .Conclusion: The current study supported that serum phosphorus, osteocalcin, β-crosslaps, TP1NP, and PTH levels were negatively associated with eGFR levels, and demonstrated a significant association between multiple measures of mineral metabolism and renal function in Chinese centenarians. Chronic kidney disease (CKD) constitutes a public health issue that is estimated to affect more than 10% of global population, and over 100 million people have CKD in China , 3. MeanCentenarians have been suggested to have a delayed or escaped onset and interaction of age-related illnesses, such as metabolic bone disease (MBD) and CKD , 10. SomMeanwhile, considering a lack of the data regarding centenarians, there is a need of studies evaluating the association between mineral metabolism and renal function in Chinese centenarians. Hainan is a longevity area with the highest population density of centenarians in China. The China Hainan Centenarian Cohort Study (CHCCS) provides a considerably population-based sample of Chinese centenarians . MoreoveCHCCS was a population-based study conducted in 18 cities and counties of Hainan Province, China, between July 2014 and December 2016. Its cohort profile has been described previously . Accordi−1.234 × age (year)−0.179 × 0.79   . DiabeteU-test. Categorical variables were reported as numbers and percentages, and compared with Chi-square test. Spearman's correlation  was used to investigate the simple correlation between mineral metabolism and renal function. Linear regression analysis was conducted following these models: model 1 adjusted for age and sex; and model 2 adjusted for age, sex, waist circumference, hypertension, diabetes mellitus, SBP, DBP, serum albumin, triglyceride, HDL-C, LDL-C, and FBG levels, so as to investigate the association between mineral metabolism and renal function. Besides, linear regression analysis was conducted following these models: model 1 adjusted for age, sex, and eGFR levels; and model 2 adjusted for age, sex, waist circumference, hypertension, diabetes mellitus, SBP, DBP, serum albumin, triglyceride, HDL-C, LDL-C, FBG, and eGFR levels, so as to investigate the association between serum PTH and 25OHD levels additionally adjusted for eGFR levels. Unstandardized β were provided and shown per 1-SD increment. As the common statistics in linear regression analyses, they were automatically and simultaneously output by Statistic Package for Social Science (SPSS) software. Statistical analyses were conducted with SPSS software version 17 , with P < 0.05 considered to be statistically significant.Continuous variables with skewed distribution were reported as medians and interquartile ranges, and compared with Mann–Whitney 2. There were 318 centenarians (42.4%) with eGFR levels <60 ml/min/1.73 m2. P < 0.05 for all) rather than serum calcium and 25OHD levels  had significant difference between centenarians with eGFR levels <60 ml/min/1.73 m2 and those with eGFR levels ≥60 ml/min/1.73 m2. In the simple correlation analyses, serum phosphorus, osteocalcin, β-crosslaps, TP1NP, and PTH levels were negatively related to eGFR levels , whereas serum calcium and 25OHD levels were not significantly related to eGFR levels . In the first and second models of linear regression analyses , whereas serum calcium and 25OHD levels were not significantly associated with eGFR levels . Serum 25OHD levels were significantly associated with serum PTH levels in the first and second models of linear regression analyses after adjusted for eGFR levels .All centenarians have a median (range) age of 102 (100–115) years, and 18.9% of these centenarians were males. Median (range) level of eGFR was 63 (16–138) ml/min/1.73 manalyses , serum pPrevious studies exploring the association between mineral metabolism and renal function have been conducted in the CKD population, among general adults and within the elderly, and as the first time, all findings from the current study provided strong support for the association in Chinese centenarians. This cross-sectional study showed that the simple correlations and multivariate associations of serum phosphorus, osteocalcin, β-crosslaps, TP1NP, and PTH levels with eGFR levels were significant, but those of serum calcium and 25OHD levels with eGFR levels were not significant in Chinese centenarians.The current study confirmed significant association between serum phosphate and renal function which was consistent with the results from previous studies , 20. CKD2), while eGFR levels were relatively maintained among these Chinese centenarians. Disordered phosphate and PTH metabolism generally precedes the change in serum calcium and 25OHD levels in the CKD population  and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. Informed consent was obtained from all participants included in the study.SF, HY, YL, FLv, JD, FZ, FLu, YZ, and YY contributed to the study design, performed the data collection and analyses, and drafted the paper.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


Heart failure threatens the lives of patients and reduces their quality of life. Heart failure, especially heart failure with preserved ejection fraction, is closely related to systemic and local cardiac persistent chronic low-grade aseptic inflammation, microvascular damage characterized by endothelial dysfunction, oxidative stress, myocardial remodeling, and fibrosis. However, the initiation and development of persistent chronic low-grade aseptic inflammation is unexplored. Oxidative stress-mediated neutrophil extracellular traps (NETs) are the main immune defense mechanism against external bacterial infections. Furthermore, NETs play important roles in noninfectious diseases. After the onset of myocardial infarction, atrial fibrillation, or myocarditis, neutrophils infiltrate the damaged tissue and aggravate inflammation. In tissue injury, damage-related molecular patterns (DAMPs) may induce pattern recognition receptors (PRRs) to cause NETs, but whether NETs are directly involved in the pathogenesis and development of heart failure and the mechanism is still unclear. In this review, we analyzed the markers of heart failure and heart failure-related diseases and comorbidities, such as mitochondrial DNA, high mobility box group box 1, fibronectin extra domain A, and galectin-3, to explore their role in inducing NETs and to investigate the mechanism of PRRs, such as Toll-like receptors, receptor for advanced glycation end products, cGAS-STING, and C-X-C motif chemokine receptor 2, in activating NETosis. Furthermore, we discussed oxidative stress, especially the possibility that imbalance of thiol redox and MPO-derived HOCl promotes the production of 2-chlorofatty acid and induces NETosis, and analyzed the possibility of NETs triggering coronary microvascular thrombosis. In some heart diseases, the deletion or blocking of neutrophil-specific myeloperoxidase and peptidylarginine deiminase 4 has shown effectiveness. According to the results of current pharmacological studies, MPO and PAD4 inhibitors are effective at least for myocardial infarction, atherosclerosis, and certain autoimmune diseases, whose deterioration can lead to heart failure. This is essential for understanding NETosis as a therapeutic factor of heart failure and the related new pathophysiology and therapeutics of heart failure. Heart failure (HF) is a complex syndrome. Its typical symptoms are breathlessness, paroxysmal nocturnal dyspnea, reduced exercise tolerance, fatigue, tiredness, increased time to recover after exercise, and ankle swelling, resulting in decreased cardiac output and/or increased intracardiac pressure . CurrentNeutrophils are powerful inducers of oxidative stress and inflammation in the immune system, but we know very little about their role and mechanism in HF. Recently, accumulating evidence shows that neutrophil extracellular traps (NETs) are an important way to be involved in the immune response. NETs are the last resort to control microbial infections released by neutrophils, and this unique cell death program of neutrophils is called “NETosis”. In this cell death process, citrullinated chromatin and bactericidal proteins from granules and cytoplasm are released and produce a network structure, which promotes the immobilization and killing of invading microorganisms in the extracellular environment. NETosis plays a vital role in host defense, autoimmunity, and blood coagulation , 15. NETTo our knowledge, the existence and function of NETs in HF, especially in HFpEF, have not been studied in detail. According to available data, in HF, especially in HFpEF, an aseptic inflammatory response with NETs as the core should be the necessary condition to enhance myocardial tissue damage, fibrosis, and ventricular remodeling.More than a decade ago, several studies have reported that an increase in circulating MPO, as an independent risk factor, may play a key role in the occurrence and maintenance of chronic HF –39. Furtα), intercellular adhesion molecule-1, collagen 1, alpha-smooth muscle actin, and transforming growth factor beta family, were significantly increased in the heart of seipin/Bslc2 knockout mice [V617F allele conditional knock-in mouse model, clonal hematopoiesis accelerated pathological HF remodeling. After exposing these mice to stressors of coronary artery ligation-induced myocardial infarction and aortic constriction-induced pressure overload, researchers demonstrated that myeloid JAK2V617F mice showed accelerated cardiac inflammation and remodeling, larger infarct size, and HF. In addition, on stimulation with LPS, THP-1 cells transduced with JAK2V617F exhibited enhanced inflammatory responses, including transcripts of IL-6, IL-1β, TNF-α, and monocyte chemoattractant protein-1 (MCP-1), as well as AIM2 inflammasome component compared with JAK2WT cells [V617F mutant neutrophils were prone to form NETs in vivo and lead to thrombosis events associated with myeloproliferative diseases. This study revealed that neutrophils expressing JAK2V617F increased the protein expression of peptidylarginine deiminase 4 (PAD4) required for NETosis and that NETosis and thrombosis in vivo driven by JAK2V617F require PAD4 [−/− mice [A recent study reported that seipin/Bslc2 knockout mice, a model of Asian lean diabetes, developed HFpEF. The authors found that increased cardiac titin phosphorylation and reactive interstitial fibrosis associated with NETs lead to left ventricular stiffness. At the age of 23 weeks, cardiac hypertrophy and diastolic dysfunction were observed. The hearts of seipin/Bslc2 knockout mice displayed an increased end-diastolic pressure/volume relationship, increased end-diastolic pressure, and left atrial enlargement as well as exercise intolerance. NET was characterized by intact neutrophils with condensed nuclei (15 weeks old) and persisted as large amorphous extracellular structures, released DNA fibers decorated with MPO, and citrullinated histone (32 weeks old). By the age of 24 weeks, the mRNA and protein levels of the markers of cardiac inflammation and fibrosis, such as interleukin-6 (IL-6), tumor necrosis factor-alpha , which lead to myocardial “no-reflow” and myocardial infarction . Multiplβ, IFN-γ, IFN-inducible protein-10, and MCP-1, patients with COVID-19 had high IL-6 levels. Echocardiography revealed that the heart was enlarged and the LVEF was significantly decreased. Bedside chest radiographs showed typical ground-glass changes indicative of viral pneumonia [β, TNF-α, and IL-8/C-X-C motif chemokine ligand 8, and infected neutrophils released MPO and triggered NETosis in the presence of TNF-α [κB activation [Recently, many case reports have described the clinical observation and pathology of “fulminant myocarditis” of coronavirus disease 2019 (COVID-19). The patients had increased levels of troponin I, creatine kinase-MB, and brain natriuretic peptide. In addition to the high concentrations of IL-1neumonia –83. Althneumonia –86. Furtneumonia , 85. COVneumonia . Coxsackneumonia , 90. In of TNF-α . Neutroptivation . Anothertivation , suggestα [Patients with diabetes have an increased risk of HF, a high prevalence of HF and hospitalization, and poor prognosis . Likewisα , 96. Patα . Anotherα . The autα . By contα , 100. Acα . Preventα . This inAccumulated evidence has confirmed that NETs are involved in the pathogenesis of autoimmune diseases such as RA, systemic lupus erythematosus (SLE), and vasculitis . In clinα, IL-6, RANTES, and IL-1ra levels [β and TNF-α in mouse cardiomyocytes [Mitochondrial DNA (mtDNA) leads to cardiac dysfunction in an aging heart. In elderly people aged >90 years, the plasma mtDNA level gradually increased with age, and the increase in the plasma mtDNA level was positively correlated with plasma TNF-a levels . Moreovea levels . Myocarda levels . Furthermyocytes . Circulamyocytes .SERCA2a gene knockout C57Bl/6J mice, 4 weeks after conditional gene knockout, the mice were randomly given the TLR-9 agonist CpG-B. After 4 weeks, administering CpG-B shortened the life expectancy of SERCA2a gene knockout mice, reduced ventricular diastolic function, and increased heart and systemic inflammation, showing exacerbation of diastolic HF [−/− and TLR9−/− mice in the area of tissue damage caused by I/R, suggesting that TLR-9 and STING pathways help mtDNA induce NET. Bone marrow neutrophils from STING−/− and TLR-9−/− mice showed a lower percentage of NET in mtDNA stimulation [NETs represent extracellular structures that bind to and kill microorganisms. However, mtDNA released from damaged tissues can also induce NETosis, which may lead to tissue inflammation and thromboembolism. A decade-old study revealed that NETosis was induced by mtDNA released from cells . mtDNA rtolic HF . Stimulatolic HF –128, promulation . These fβ secretion. The released IL-1β stimulates the autonomous granulocyte production of hematopoietic stem cells in the bone marrow, which requires NETosis [In addition to mtDNA, some other damage-associated molecular patterns (DAMPs) are used as important biomarkers and proinflammatory factors of HF, such as S100A8/A9 , high mobility box group box 1 (HMGB1), fibronectin extra domain A (FN-EDA), and GAL3 , 130. Mu NETosis , 137.−/− mice, FN-EDA promoted myocardial I/R injury, increased infarct size, elevated plasma cardiac troponin I levels, induced neutrophil infiltration and extracellular traps, and caused cardiomyocyte apoptosis [In addition to the Kubota group, which used HMGB1 transgenic mice to show that cardiac nuclear HMGB1 exerts protective effects on myocardial infarction, cardiac hypertrophy, and HF –140, manpoptosis . GAL3 ispoptosis , 153, aspoptosis , 155. Thpoptosis , 157. GApoptosis , 157. Inpoptosis –160. Othpoptosis –164. HMGpoptosis , 166. Mopoptosis . In myocpoptosis .β2 integrins during acute inflammation [γRI (CD64) and IgG FcγRII (CD32) high-affinity receptors [β, NETosis, and NET-associated tissue factor plasma levels [Dilated cardiomyopathy caused by myocarditis can develop into HF. A recent study reported that midkine induced cardiac inflammation by promoting neutrophil trafficking and NETosis in myocarditis. In EAM in mice, midkine as a target can not only inhibit NETosis and neutrophil infiltration in vivo but also reduce myocardial fibrosis and maintain cardiac function . Midkineammation –178. Morammation . The neuammation . The plaammation . Compareammation . Recent ammation . Thirty ammation . Then, ieceptors , 185. Rea levels . The sera levels . Earliera levels –190. Besa levels , 192. Rea levels . The aboγ [As we all know, oxidative stress is closely related to inflammation. A large amount of evidence has shown that inflammation plays an important role in the onset and development of cardiovascular diseases . Similarγ  , a 16-carbon chain chlorinated fatty aldehyde, accumulated in the hearts of rats with left anterior descending artery occlusion. Compared with sham-operated rats, levels of 2-ClHDA and neutrophil infiltration in myocardium of surgical infarction rats were increased [Recently, a study reported that the redox imbalance of neutrophils during the oxidative burst promotes the generation of NETosis. The authors believed that the generation of NETosis is partly regulated by the changes in the cytosolic thiol redox homeostasis in neutrophils and depends on the circumstances under which NETosis is generated . Meanwhid carbon . Becaused carbon . In the ncreased . In recencreased   . Low serth HFpEF . In patith HFpEF . Recentlth HFpEF . The freth HFpEF . These s+/+ wild-type control group, the thrombotic occlusion time in Sirt3−/− mice was reduced by half [ex vivo LPS-induced NETosis was increased in Sirt3−/− bone marrow-derived neutrophils, indicating that SIRT3 deletion in the inflammatory environment affected NETosis and arterial thrombosis in mice [SIRT3 is a major mitochondrial deacetylase and an important factor in maintaining cardiac mitochondrial bioenergy. Multiple studies have revealed that SIRT3 deficiency or decreased expression can induce the excessive acetylation of pyruvate dehydrogenase, ATP synthase, cyclophilin D, and mitochondrial dynamic like GTPase OPA1; increase the opening of mitochondrial permeability transition pore; and change the process of oxidative phosphorylation, resulting in mitochondrial dysfunction –228. Fur by half . Moreove in mice . Another in mice .In recent years, several studies have shown that NET promotes thrombus formation , 232. Ne−/− mice showed prolonged time for thrombosis and complete occlusion, and the thrombus growth rate was significantly reduced compared with FN-EDA+/+ mice. TLR-4 gene deletion reversed accelerated thrombosis in FN-EDA+/+ mice [+/+ mice, galectin-3-binding protein (GAL3BP) and GAL3 were localized in the vein wall, red blood cells, and platelets, whereas only GAL3 was expressed in neutrophils. GAL3 increased significantly during early DVT, and GAL3BP/GAL3 was co-located at the interface between neutrophils and endothelial cells. Some neutrophils attached to the vein wall, whereas the number of activated neutrophils in the vein wall of GAL3−/− mice significantly decreased. Thrombus size was associated with increased levels of GAL3 and IL-6 in the venous wall [Recent studies have revealed that DMAPs that induce NETs also promote thrombosis, such as HMGB1, FN-EDA, GAL3, and C-X-C motif chemokine receptor 2 (CXCR2). In addition to activating neutrophils, HMGB1 also regulates platelet activation, granule secretion, adhesion, and diffusion through the TLR-4 pathway on platelets . Similar+/+ mice . Althougous wall . IL-8, nous wall . Neutropous wall . The posous wall , and theous wall . In clinous wall , and manous wall .− ions. However, MPO is also associated with oxidative stress and tissue damage resulting from the potent oxidant HOCl, which plays a key pathogenic event in a variety of inflammatory states, including cardiovascular disease. MPO inhibitors, such as KYC, seem to prevent PMA-stimulated neutrophil HOCl without affecting superoxide production [Neutrophil specific enzymes, MPO, PAD4, and NE, play a key role in the formation of NETs. Up to now, PAD4 inhibitors such as BMS-P5, GSK484, GSK199, and cl-amidine and MPO inhibitors such as PF-1355, PF-06282999, INV-315, and AZD5904 have been developed. MPO is a peroxidase containing heme, mainly expressed in neutrophils and a small amount in monocytes, where it catalyzes the production of reactive oxygen intermediates such as hypochlorous acid (HOCl) as antibacterial armory from hydrogen peroxide and Cloduction . Anotheroduction . PAD4 anoduction .−/− atherosclerosis mouse model. Treatment with Cl-amidine for 11 weeks decreases the atherosclerotic lesion area, reduces the recruitment of neutrophils and macrophages to the artery and prevents the formation of NET, and downregulates interferon-α expression in the arteries. In addition, Cl-amidine treatment delays carotid artery thrombosis in a photochemical injury model [−/− mouse model of atherosclerosis [Recently, several studies have reported the effect of PAD4 inhibitors on cardiovascular disease . In a mory model . Howeverclerosis . Likewisclerosis . Furtherclerosis . Moreoveclerosis , 258. Siclerosis , while Sclerosis –263. Theclerosis .Statins and metformin may effectively improve heart failure clinically (see review article References –268). InIn short, DAMPs, such as mtDNA, HMGB1, FN-EDA, and GAL3, released by damaged organs may activate neutrophils and form NETs through TLR-4/9 and RAGE, which requires the participation of oxidative stress, thereby causing or aggravating cardiac inflammation and fibrosis. NETs promote thrombosis of coronary microvessels and affect cardiac function. In some studies, neutrophil-specific MPO and PAD4 ablation or inhibition could improve the disease progression of HF, myocardial infarction, and atrial fibrillation, which all illustrate the fact that NETosis is a pathogenic factor of HF. Recently, Abrams et al.  have dev


Although their roles in cancer treatment and prevention have been extensively reported, interestingly, they may also have distinctive properties in energy metabolism and metabolic disorders. We review inositols and inositol phosphate metabolism in mammalian cells to establish their biological activities and highlight their potential roles in energy metabolism. These molecules are known to decrease insulin resistance, increase insulin sensitivity, and have diverse properties with importance from cell signaling to metabolism. Evidence showed that inositol phosphates might enhance the browning of white adipocytes and directly improve insulin sensitivity through adipocytes. In addition, inositol pyrophosphates containing high-energy phosphate bonds are considered in increasing cellular energetics. Despite all recent advances, many aspects of the bioactivity of inositol phosphates are still not clear, especially their effects on insulin resistance and alteration of metabolism, so more research is needed.Recently, inositols, especially  Myo-inositol (myoIns) and inositol hexakisphosphate (IP6 or InsP6 or phytic acid) are common in biology; these naturally occurring carbohydrates are widely distributed among plants and mammalian cells, with multiple roles -inositol incorporation into inositol phosphates elicited different response than the phosphatidylinositol phosphates during the development of Dictyostelium discoideum [Although much of what we know about the inositol phosphates metabolism was studied in  pigeons . This fi pigeons , suggestscoideum . The incscoideum .myo-inositol 1,4,5-trisphosphate, and phosphoinositides, is well documented in biological processes, myoIns and the other inositol phosphates also possess biological activities [Although signaling via inositol phosphates, e.g., the second messenger tivities . It is wtivities ,24. Hightivities . Likewistivities . These htivities  that britivities , RNA exptivities ,36, mRNAtivities , DNA doutivities ,38, genetivities ,40,41, ptivities ,43, and tivities , some ofRNA editing. As mentioned above, the enzymes IMPK and IP2K, which are necessary to synthesize IP6 from its precursor, are concentrated in the nucleus, suggesting a role for IP6 in nuclear function in human cells. Indeed, crystal structures have revealed interactions of IP6 with various regulatory elements through electrostatic interaction with proteins containing highly basic residues. Macbeth et al., reported in 2005 that IP6 binds to the core catalytic domain at the core of adenosine deaminase (ADAR2), an RNA editing enzyme that converts adenosine (A) to inosine (I) [sine (I) .RNA export and gene expression. In eukaryotes, messenger RNA needs to be processed to messenger ribonucleoprotein (mRNP) in order to be exported from the nucleus to the cytoplasm through nuclear pore complex (NPC), which requires the action of the DEAD-box ATP-dependent RNA helicase DDX19. However, the recruitment and interaction of nucleoporins Nup42 and Gle1 is necessary for activity [IPPK gene revealed selectivity for a subset of RNA involved, including those that affect cell cycle G1/S checkpoint regulation and inflammatory response [activity . It has activity ,46. Furtresponse . Anotherresponse . Howeverresponse . In addiresponse , suggestresponse ,48. In cresponse ,40,41.Phosphate homeostasis. Using CRISPR to completely disrupt IP6K1 and IP6K2 genes in a human colon carcinoma HCT116 cell line, it was shown that although there was an increase in the amount of ATP and intracellular free phosphate, these cells had limited ability to uptake radioactive phosphates [osphates . This suosphates . Since Xosphates ,49,50.The prevalence of diabetes mellitus has been growing worldwide with an estimated to affect 9.3% or 463 million people in 2019 which will be increasing to 10.2% or 578 million by 2030 and 10.9% or 700 million by 2045 . Diabeted-chiro-inositol decreased serum glucose in diabetic rats [d-chiro-inositol have been shown to counteract metabolic disorders [In 1986, the role of inositols in insulin signaling pathway was demonstrated by a research group showing that insulin and phosphatidylinositol-specific phospholipase C enzyme (PLC) synergistically increased cyclic nucleotide (cAMP) phosphodiesterase (PDE) activity in fat cells of rats . PDE is tic rats . This evisorders  and imprisorders ,64,65.d-chiro-inositol, or myoIns alone improved lipid profiles including decreased plasma low density lipoprotein (LDL) and triglycerides and increased plasma high density lipoprotein (HDL) [d-chiro-inositol decreased insulin levels after oral glucose tolerance test (OGTT) in obese human subjects [MyoIns is the most common isoform shown to have therapeutic effects in human health including metabolism, reproduction, cell growth and survival, and development of nervous system . Moreovein (HDL) , decreasin (HDL) , reducedin (HDL) ,63,66,67in (HDL) . Furthersubjects , suggestsubjects ,69. Its subjects . Targetisubjects , metabolsubjects , and PCOsubjects ,70. A mesubjects . Interessubjects .glucose transporter type 4 (GLUT4) mRNA, insulin receptor substrate 1 (IRS-1) and phosphorylated insulin receptor substrate 1 (p-IRS-1) mRNA, decreased basal lipolysis, and increased adipocyte differentiation in 3T3L-1 mouse adipocyte [phosphoenolpyruvate carboxykinase (PEPCK) gene, which produces an essential enzyme in gluconeogenesis, after the activation of 8-bromo-cAMP in hepatocyte of rats [IP6 has been shown to reduce blood glucose and delay carbohydrate digestion and absorption after supplementation in humans . A studydipocyte . This ev of rats . In addi of rats . An expe of rats . IP6 als of rats .It is well known that obesity is now considered as a major health risk globally. People worldwide are facing obesity and/or overweight status, especially in the United States. The prevalence of obesity in adults was over 30% in 2015, and is rising over time . ObesityIP6K knockout displayed reduced body weight, fat accumulation, and percentage of fat mass as well as increased percentage of lean body mass [IP6K knockout also improved metabolic parameters, including blood glucose [IP6K1 did not increase phosphorylated Akt in muscle and glycogen synthase kinase 3β (GSK3β), a rate-limiting enzyme promoting deposition of glycogen [IP6K1 ablation might be associated with reduced Akt pathway.The effects of inositol phosphates and IP6Ks have long been considered for a target of obesity and metabolic diseases . Many exody mass ,81, sugg glucose ,21, totaglycogen , in whitIP6K1 knockout in adipocytes increased oxygen consumption rate and adipocyte browning genes [uncoupling protein 1 (UCP1), peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1α), PR domain containing 16 (PRDM16), and peroxisome proliferator-activated receptor alpha (PPARα), indicating that IP6K1 deletion might enhance the browning of adipocytes [IP6K1 deletion was also shown to increase energy expenditure and lipid oxidation via the AMP-activated protein kinase (AMPK) in mice fat cells and decreased fatty acid synthesis in 3T3-L1 adipocytes [IP6K1 knockout mice, energy intake was not reduced, whereas body weight decreased, while oxygen consumption and energy expenditure increased, when compared with wild-type control, suggesting that IP6K1 is a dominant player in energy output [IP6K1 knockout (HFD-AdKO) mice fed with high fat diet showed improved fatty liver than their counterpart [IP6K1 might have the advantages in decreasing obesity, improving metabolic parameters, and increasing thermogenic energy metabolism. In other words, type 2 diabetes, obesity, and non-alcoholic fatty liver might be ameliorated by IP6K1 deletion.It has been known that the browning of white adipocytes leads to increased cellular thermogenesis resulting in an increase in energy expenditure ; this prng genes  includinipocytes . Indeed,ipocytes . In IP6Ky output . In addinterpart . Taken td-chiro-inositol have been shown to reduce risks of metabolic diseases, including diabetic mellitus, dyslipidemia [d-chiro-inositol could be transformed from myoIns by the inversion of C3 hydroxyl via insulin-dependent epimerization [chiro-inositol include soybeans, legumes, oranges, arrowroot, and ginseng [d-chiro-inositol and IP6K also brought attention to the role of inositol phosphates in metabolism.The association between inositols and metabolism has been linked by many research groups. Among inositol isomers, the myoIns and ipidemia ,63,64, aipidemia . d-chirorization . Natural ginseng . Moreovedb/db mice (type 2 diabetes mice) revealed that d-chiro-inositol improved glucose levels and significantly increased hepatic glycogen when compared with the control group [d-chiro-inositol also increased the protein expressions of insulin receptor substrate 2 (IRS2), PI3K, Akt, GLUT4, and phospho-Akt but decreased GSK3β protein in hepatic cells [d-chiro-inositol improved glucose metabolism and hepatic glycogen synthesis through the upregulation of insulin receptor, GLUT4, GSK3β, and PI3K-Akt cascades. A previous study showed that, for mice in fasting state, glucagon stimulated PKA leading to phosphorylation of inositol 1, 4, 5-trisphosphate receptors (InsP3Rs) with an increase in Ca2+ in the cytosol and activation of CREB-regulated transcription coactivator 2 (CRTC2) which is associated with hepatic glucose production [IP6K1 deletion might turn off the program of insulin action which activates the Akt pathway in stimulating glucose uptake, glycogen synthesis, and protein synthesis [As mentioned above, 5PP-IP5 or IP7 acts as an energetic molecule and IP6K1 is a key enzyme to produce inositol pyrophosphate IP7 from IP6 ,15. A prol group . Additioic cells . This fioduction . Whereasoduction . These dynthesis .Furthermore, a previous study showed that inositol polyphosphate multikinase (IPMK) and IP6K1 played a critical role in the cascade of inositol phosphates synthesis and have an impact on energy metabolism in mammals . To illuIP6K1 appeared to increase basal lipolysis by modulating protein perilipin1 (PLIN1) in 3T3L1 adipocytes suggestive that IP6K1 and inositol pyrophosphate biosynthetic process have an impact on the regulation of lipid metabolism [IP6K3 mRNA in mice and humans has the highest expression in skeletal muscle when compared with other inositol kinase family members, e.g., ITPK1, IMPK, IP6K1, and IP6K2, and its expression in muscle was high in diabetic and fasting conditions of mice [IP6K3 knockout mice showed decreased body weight, fat mass, blood glucose, blood insulin, plasma lactate, and increased glucose tolerance from age induced obesity [IP6K1 gene ablation on alterations of metabolism in metabolic target tissues including liver and adipose tissues. However, its effects on skeletal muscle, e.g., glucose uptake and/or energy expenditure as well as molecular signaling pathways, need to be elucidated. Here, we summarize the effects of myoIns, d-chiro-inositol, and IP6 on metabolic alterations in For the role of inositol phosphates on lipid metabolism, deletion of tabolism . PLIN1 itabolism . Under ttabolism . In additabolism . A previ of mice , suggest obesity . So, IP6scyllo-inositol administered orally for 1 month improved spatial memory, and decreased plaque and amyloid-β peptides (Aβ) aggregation [d-chiro-inositol was able to enhance the ability of insulin to protect central nervous system (CNS) synapse damage caused by the accumulation of toxic Aβ oligomers in association with Alzheimer’s disease [Deregulation of the inositol phosphates metabolism has been recognized in several illnesses—mostly in animal models, including neurological disorders , PCOS 7, metabolregation , suggestregation . Moreove disease . IP6 has disease . In Alzh disease . IP6 sig disease .MyoIns has been used for years against depression and anxiety disorders in patients with depressive disorder ,105,106.Interestingly, myoIns either alone or in combination with selenium can have beneficial effects in mice exposed to cadmium ,110,111.Although advanced health-beneficial effects of inositol phosphate have now been investigated, cancer preventive and therapeutic properties of IP6 have received most attention and its broad-spectrum of anticancer activities has been shown in multiple preclinical experimental studies and in humans, alone or in combination with myoIns ,27,112, IP6K1 knockout BMMSCs reduced adipocyte differentiation and enhanced osteocytes [Another interesting line of studies indicated that the consumption of IP6 can prevent development of osteoporosis and had a protective effect against osteoporosis . IP6 wasteocytes . This evtumor necrosis factor alpha(TNF-α), interleukin 6(IL-6), and interleukin-I beta(IL-1β) in liver tissues of iron overloaded induced liver injury mice [d-myo-inositol supplement both orally (400 mg/kg) and intraperitoneally (200 and 800 mg/kg) suppressed inflammation in rats with adjuvant disease [myo-inositol-1-phosphate synthase (Ino-1) deletion in bacteria (Actinobacteria Corynebacterium glutamicum) increased production of ROS and decreased cell viability suggested that Ino-1 might have an effect on oxidative stress resistance [Furthermore, anti-inflammatory effects of IP6 and myoIns was shown ; IP6 redury mice . d-myo-i disease . Moreovesistance . Becausesistance .d-chiro-inositol, both insulin-sensitizing agents, in assisted reproductive treatment (ART) has been suggested, because of the known beneficial effects of these inositols in both female and male reproduction [Moreover, the prevention of kidney stones and other pathological calcifications, such as sialolithiasis, a common disease of salivary glands, and cardiovascular calcification, that frequently occurs in the heart vessels, has been known for IP6 . Moreoveoduction .d-chiro-inositol, and IP6 have been shown to consistently decrease insulin resistance and to improve insulin sensitivity. However, the effects of inositol phosphates on carbohydrate and glucose metabolism are still poorly understood.Indeed, inositols and the inositol phosphates have been shown to have diverse health benefits such as anticancer, anti-diabetic, anti-oxidant, and anti-inflammation ,27,69. MThe complex interaction among inositol molecules, insulin signaling, and carbohydrate and/or glucose metabolism pathways would be intriguing for further studies. We would hypothesize that inositol phosphates might alter the actions of second messenger molecules in the energy metabolism pathways of insulin-sensitive tissues including liver, muscle, and adipose tissue. The better understanding on the interplay of these molecules in insulin-sensitive tissues might explore novel knowledge and provide treatment options to treat patients with metabolic syndrome. Interestingly, discussing biological regulation systems for metabolic networks classification, when a new metric system is proposed for comparisons of different metabolic systems, inositol and its derivatives involved in membrane signaling are featured as a simple molecular system offering new perspectives .Lastly, because inositols and the inositol phosphates, are abundant in food sources, especially in the Mediterranean diet , their r


Adrenal insufficiency (AI) is a potentially life-threatening condition that can be difficult to diagnose, especially if it is not considered as a potential cause of a child's clinical presentation or unexpected deterioration. Children who present with AI in early life can have signs of glucocorticoid deficiency , mineralocorticoid deficiency , adrenal androgen excess , or associated features linked to a specific underlying condition. Here, we provide an overview of causes of childhood AI, with a focus on genetic conditions that present in the first few months of life. Reaching a specific diagnosis can have lifelong implications for focusing management in an individual, and for counseling the family about inheritance and the risk of recurrence. Adrenal insufficiency (AI) is a potentially life-threatening condition that needs urgent diagnosis and treatment –4. AI issecondary causes, due to disruption of hypothalamic or pituitary (corticotrope) ACTH release, and primary causes, which affect the adrenal gland itself. Although some conditions have fairly typical presentation patterns and ages of onset, there is often a spectrum of features, and milder variants may produce partial or delayed onset forms of classic conditions  should not be overlooked.Secondary AI is caused by impaired ACTH synthesis and release from pituitary corticotrope cells . ACTH deGLI1, HESX1, LHX3, LHX4, SOX3, SOX2 and others) . Disruption of PROP1 or GH1 can cause ACTH insufficiency in later life  . Pituita others) . Other ater life .TBX19), or with associated features due to defects in pro-opiomelanocortin (POMC) or pro-hormone convertase-1 (PC-1/PCSK1).Isolated ACTH deficiency can occur due to disruption of TPIT  . POMC isDefects in POMC itself also result in ACTH insufficiency and adrenal dysfunction in early infancy . ChildrePCSK1) also presents with ACTH insufficiency, together with hypoglycemia, malabsorptive diarrhea, obesity, and hypogonadism  in childhood is shown in DHCR7) (Smith–Lemli–Opitz syndrome is a defect in cholesterol biosynthesis due to disruption of the enzyme 7-dehydrocholesterol reductase (DHCR7) . Common DHCR7) . AI or iDHCR7) , 22. EleSTAR) plays a key role in cholesterol transport into the mitochondria, whereas the P450 cholesterol side-change cleave enzyme  catalyzes the conversion of cholesterol to pregnenolone , 24STAR)nenolone . In contnenolone , 27. Parnenolone .CYP21A2) is most prevalent, with an incidence of 1:10,000–1:20,000  (CYP11B1) , 3 beta-hydroxysteroid dehydrogenase deficiency (HSD3B2), 17 alpha-hydroxylase deficiency (CYP17A1) and P450 oxidoreductase deficiency (POR), all of which have specific presentations and biochemical profiles  . In virts occur) , 33. Othprofiles  23)..CYP21A2)Approximately 80% of 46,XX girls with confirmed 21-OHD have atypical genitalia at birth, so any baby with genital differences and non-palpable gonads should be considered as having 21-OHD until proven otherwise , 34. ProAdrenal hypoplasia is an underdevelopment of the adrenal glands, which often presents with PAI in early life . UsuallyNR0B1)  primarily affects boys and is associated with disruption of the nuclear receptor, DAX-1 (encoded by NR0B1) , 36. ThiNR0B1) . Late-onNR0B1) –40.macrophallia at birth, and initial presentation with either isolated mineralocorticoid insufficiency or isolated glucocorticoid insufficiency is reported  in adolescence, and impaired spermatogenesis . Some boreported , 43. Groreported , 44–46.NR0B1, around one-sixth have a localized deletion of this gene on the X-chromosome (Xp21), and one-sixth have a larger Xp contiguous gene deletion syndrome that can involve genes causing glycerol kinase deficiency (GKD), ornithine transcarbamylase deficiency (OTC) and Duchenne Muscular Dystrophy (DMD)  . Very rahy (DMD) . Establihy (DMD) . Familiehy (DMD) .NR5A1) is a related nuclear receptor considered as a “master-regulator” of adrenal and reproductive development   , 52–54. intrauterine growth restriction, metaphyseal dysplasia, adrenal hypoplasia, genitourinary anomalies)  , 56. Chide novo. IMAGe-associated pathogenic variants are localized within a very specific motif in the PCNA-binding motif of CDKN1C, causing impaired cell cycle S-phase progression , often involving a heterozygous intronic variant (c.1686 + 32C>G). POLE1 is a DNA polymerase that interacts with PCNA in S-phase DNA replication.Recently, an “IMAGe-like” syndrome with AI and immunodeficiency  has been reported . These cmyelodysplasia, infections, restriction of growth, adrenal hypoplasia, genital phenotypes, enteropathy)  , 62. InfSAMD9)  , 62. Thecis  “removes” the deleterious gain-of-function mutation in SAMD9 allowing a clonal growth advantage of these affected cells, especially in the hematopoietic system, and reversal of the postnatal anemia and thrombocytopenia (loss-of-function (nonsense or frameshift) changes or uniparental disomy in cis can also “remove” the mutant allele and ameliorate the phenotype  has been reported in a single family with homozygous disruptive mutations in WNT4, a signaling molecule implicated in adrenal development , pseudotrisomy 13, Galloway–Mowat syndrome (WDR73), Pallister–Hall syndrome  and Meckel–Gruber syndrome (MKS1)  .Another important group of conditions causing PAI in childhood are ACTH-resistance conditions  and related disorders . Some ofMC2R)  is a recessive condition that results from pathogenic variants in the ACTH receptor  , 69. ChiMC2R) , 71. FGDA similar form of ACTH-resistance results from disruption of the melanocortin 2 receptor accessory protein, MRAP . MRAP trNNT) are a well-established cause of isolated PAI in children, and occasionally additional features such as early puberty have been reported (TNXRD2) are reported in a single family (sometimes with cardiac defects), and present in mid- or later childhood , a potential nucleoporin component that may also influence cellular stress (Triple A syndrome is a well-established condition linking PAI (“Addison disease”), with alacrima and achalasia of the esophagus , 79. Thir stress –82. Alacr stress , 84. ProMCM4) causes mild PAI together with short stature and immunodeficiency (STAR and CYP11A1) can present in childhood with PAI. Non-classic STAR defects are sometimes termed FGD3  and has many features including hypotonia, seizures, hepatic dysfunction and renal cysts. PAI has been reported, usually in childhood or with an impaired stress response (Adrenoleukodystrophy (ALD) is a very important cause of PAI because of associated progressive neurological features . The X-lstigated –94. In cstigated . This spresponse , so screresponse .MK-TK, MRPS7, QRSL1, NDUFAF5, GFER) , but also pathogenic variants in other related genes  , 97.LIPA), and is associated with AI , failure to thrive, hepatosplenomegaly and anemia in the first few months of life. It is a lysosomal storage disorder that is usually fatal, although improvements with enzyme replacement treatment  are reported (Wolman disease (primary xanthomatosis) results from disruption of lysosomal acid lipase (reported –100.AIRE). Early features can include mucocutaneous candidiasis and rarely hypoparathyroidism . PAI and other associations usually occur in childhood or later life.Although autoimmune “Addison disease” is the most common cause of AI in adolescents and adults, autoimmune PAI is rare in children . The besPhysical causes of AI such as hemorrhage or infiltration  should not be overlooked. Unilateral adrenal hemorrhages detected by imaging are common (1:200–500 newborn), but usually asymptomatic . SymptomTransient, relative AI has been described in some very preterm babies, or in sick newborn children under stress. The physiological basis of this is unclear, but steroid supplements have been used in some situations .Making a specific genetic diagnosis has several benefits. It allows tailored treatment of the specific underlying hormonal defect  and permits the surveillance, early recognition, and prompt treatment of associated extra-adrenal features , 87, 98.de novo; X-linked, imprinted). This information guides genetic counseling during future pregnancies, and potentially allows pre-symptomatic diagnosis and treatment in relatives with subclinical disease , where there is a specific biochemical profile, well-established pathogenic variants, and a pseudogene that can complicate analysis, or in X-linked AHC (DAX-1/NR0B1) when well-established associations  or inheritance patterns  are present.Traditionally, genetic testing has relied on Sanger sequencing of However, associated features or pathognomonic biochemical patterns are often not present when an infant presents with PAI, so “next generation” sequencing (NGS) approaches are increasingly time- and cost-effective.targeted “panels” to analyze all key PAI genes at once are increasingly available as a clinical service. In addition, several studies have shown how “trio” whole exome sequencing (WES) can help diagnose sick infants and children, especially when there are complex, multisystem features, and exome analysis has been reported to help in the diagnosis of children with PAI (Whole genome sequencing (WGS) will become increasingly available and has potential advantages and disadvantages at present compared to panels/WES.Access to services varies from country to country, but with PAI . Whole gIn general, genetic testing for PAI has a high diagnosis rate, certainly when compared to other pediatric endocrine conditions such as congenital hypothyroidism and hypothalamo-pituitary hormone deficiencies. For example, in a national cohort study of rare, undiagnosed PAI in Turkey (with CAH and obvious metabolic causes excluded), a specific genetic diagnosis was reached in 80–90% of children , althougMRAP splice variant cIVS3ds + 1delG is found in the West, whereas P450scc/CYP11A1 p.R451W occurs in Eastern regions (Finally, founder effects and genetic “hotspots” can be very important in identifying a specific genetic cause of PAI and taking a history of family ancestry is key. For example, in Turkey the  regions . As famiAI is an important diagnosis to consider in any sick newborn infant and prompt investigation and treatment is essential. Genetic testing is increasingly useful for finding a specific cause, predicting associated features, counseling families and, in some situations, for modifying treatments.All authors were involved in the writing, editing, and final approval of the manuscript.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


The specific perception of ABA by PYR1 represented a regulatory node, modulating different outcomes in disease resistance.ABA is involved in plant responses to a broad range of pathogens and exhibits complex antagonistic and synergistic relationships with salicylic acid (SA) and ethylene (ET) signaling pathways, respectively. However, the specific receptor of ABA that triggers the positive and negative responses of ABA during immune responses remains unknown. Through a reverse genetic analysis, we identified that PYR1, a member of the family of PYR/PYL/RCAR ABA receptors, is transcriptionally upregulated and specifically perceives ABA during biotic stress, initiating downstream signaling mediated by ABA-activated SnRK2 protein kinases. This exerts a damping effect on SA-mediated signaling, required for resistance to biotrophic pathogens, and simultaneously a positive control over the resistance to necrotrophic pathogens controlled by ET. We demonstrated that PYR1-mediated signaling exerted control on a priori established hormonal cross-talk between SA and ET, thereby redirecting defense outputs. Defects in ABA/PYR1 signaling activated SA biosynthesis and sensitized plants for immune priming by poising SA-responsive genes for enhanced expression. As a trade-off effect,  Pathogen recognition triggers the altered accumulation of three major defense hormones: salicylic acid (SA), jasmonic acid (JA), and ethylene (ET). SA is essential for establishing resistance to many virulent biotrophic pathogens, especially as a component of systemic acquired resistance (SAR) ,2, while2 [ABA is another major phytohormone involved in the regulation of a great variety of abiotic stress responses in plants. In addition, ABA assists in controlling many developmental and growth characteristics of plants, including seed germination and dormancy, leaf abscission, closure of stomata, or inhibition of fruit ripening . ABA als2 .Three major protein families form the core ABA signaling pathway; (i) the soluble ABA receptors, which are 14 members of pyrabactin resistance 1 (PYR1) and PYR1-like (PYL) proteins, also known as regulatory component of ABA receptors (RCAR) family and collectively referred to as PYR/PYL/RCAR, (ii) group A of type 2C protein phosphatases (PP2Cs), and (iii) SNF1-related protein kinases (SnRKs) subfamily 2 (SnRK2s), namely SnRK2.2, 2.3 and 2.6  ,36,37,38pyr/pyl mutants shows quantitative regulation of both stomatal aperture and transcriptional response to ABA [PYR1, PYL1, PYL2, PYL4, PYL5, and PYL8, generates a mutant that is practically blind to ABA in the classical assays that measure ABA sensitivity [pyl8 mutant has been reported to show a non-redundant role in root sensitivity to ABA [pyr1 shows wild-type sensitivity to ABA and only shows a conditional phenotype -pyrabactin resistance in germination assays in medium supplemented with the ABA agonist pyrabactin [The PYR/PYL/RCAR ABA receptor family is unusually large, comprising 14 members in Arabidopsis and even more in crops, such as tomato, maize, or soybean  ,46,47. He to ABA . Inactivsitivity . Howeversitivity ,48,49, oy to ABA . In contrabactin . In eukaPlectosphaerella cucumerina  is central to ABA signaling [P. cucumerina or mock-treated, and leaf samples were collected at 0, 24, and 48 h post-inoculation (h.p.i.). Immunoprecipitation of SnRK2.2-HA and SnRK2.6-HA and the subsequent kinase assay of the immunoprecipitate were performed by determining the incorporation of 32P to purified ABF2 protein fragment substrate (amino acids Gly-73 to Gln-119) [Liquid chromatography-mass spectrometry (LC-MS) showed marked accumulation of ABA in full expanded leaves of Arabidopsis plants at 72 h after drop inoculation with a spore suspension of the fungal necrotroph cumerina A. ABA enn factor    in gel-kP. cucumerina. Symptoms of the fungal disease appear in the form of necrotic lesions, which are measured to quantify the degree of plant susceptibility [P. cucumerina when compared to Col-0 plants  was similarly affected in disease resistance to this pathogen  is required for other aspects of plant immunity [ABA signaling through SnRK2s is negatively regulated by clade A protein phosphatase type 2C (PP2C), particularly by ABI1, ABI2, PP2CA/AHG3, AHG1, HAB1, and HAB2  exhibited noticeably enhanced disease susceptibility  enhanced disease susceptibility to P. cucumerina. The results showed that the PYR1 receptor was pivotal for eliciting ABA-mediated defense responses towards P. cucumerina. We next investigated which one of 14 soluble PYR/PYL/RCAR receptors perceived the ABA produced during y to ABA ,46. Addiy to ABA . We chart growth , were intibility ), to ocpsistance ), and Cotibility A, which pyr1-1 mutant. The individual pyr1-1 mutant had a compromised disease resistance phenotype  for which resistance to the fungus remained intact. Moreover, the overexpression of the PYR1 receptor (PYR1-OE line) conferred significant enhancement of resistance to the fungus , consishenotype C. These PYR1 expression in response to the pathogen. Therefore, we next investigated whether transcriptional reprogramming occurred to enhance PYR1 expression upon pathogen inoculation. Transgenic plants expressing the promoter of the PYR1 gene fused to the β-glucuronidase GUS reporter gene (pPYR1::GUS) [P. cucumerina-mediated activation of PYR1. Transgenic lines carrying the pPYL1::GUS and pPYL4::GUS gene constructs were also assayed to determine specificity. Local infection, i.e., by drop inoculation on the upper leaf surface with a P. cucumerina spore suspension, of transgenic pPYR1::GUS plants revealed early transcriptional activation of PYR1 triggered by the pathogen   were usepathogen A. PYR1 imstances A. The lo callose B and lat callose C. These nfection D, thus sP. syringae DC3000 has been previously documented [P. syringae DC3000 could similarly be funneled through PYR1. If so, we would expect resistance enhancement in pyr1 plants. pyr1 plants were inoculated by leaf infiltration with P. syringae DC3000, and the rate of bacterial growth in the inoculated leaves was determined at 3 days post-inoculation in comparison to aba2, pyl1, pyl4, and Col-0 plants. aba2 and pyr1 mutants compared to Col-0, pyl1, and pyl4. This result confirmed the negative role of ABA in resistance towards P. syringae DC3000 and demonstrated the specific requirement of PYR1 for the negative role of ABA during this plant-pathogen interaction. Moreover, pre-treatment of Col-0 with 150 μM ABA, applied by drenching, predictably provoked disease susceptibility enhancement to P. syringae DC3000  biotrophic pathogens cumented ,19,20,21e DC3000 B, denoti hormone B.Therefore, our results indicated that the dual antagonistic role of ABA in plant immunity was mediated through the PYR1 receptor, which reciprocally activates and represses immune responses towards necrotrophic and biotrophic pathogens, respectively.pyr1 and aba2 plants carried constitutive elevated expression of SA-responsive genes, which might explain the observed enhanced resistance to P. syringae DC3000 . Western blot analysis of protein extracts derived from healthy Col-0 and pyr1 plants showed positive immunoreactive signals in two polypeptides corresponding to MPK6 and MPK3   , and the4 h.p.i. . At the ime zero . This re RT-qPCR A,B. Inte0 plants .aba and pyr1 plants exhibit enhanced disease resistance to P. syringae DC3000. These observations support that ABA and PYR1 function as a repressor module of SA-mediated onset of resistance.Thus, we hypothesize that the lack of ABA perception through the PYR1 receptor de-represses a pathway that allows cell sensitization through MPKs activation and downstream defense gene reprogramming, even in the absence of pathogen infection. Sensitized cells may be ready for the enhanced induction of this defense pathway following pathogen infection, which, in turn, may explain why pyr1 plants. The expression of the extracellular subtilase SBT3.3 gene has been recently described to be a switch for poising SA-related gene expression and immune priming [SBT3.3 expression, MPK activation, and readied SA-related genes convey in plants defective in the RNA-directed DNA methylation (RdDM) pathway, which negatively regulates immune priming [SBT3.3 and concurrent downregulation of NRPD2 in pyr1 plants compared to Col-0, congruent with the activation of immune priming in the mutant. The downregulation was specific for NRPD2, encoding the second large subunit of Pol V, because the expression of the gene encoding the large NRPE1 subunit exhibited a minimal variation in pyr1 plants  to analyze H3K4me3 and H3K9ac activation marks on the SBT3.3 and PR-1 gene promoter regions in pyr1 and Col-0 plants. We also examined the genes encoding WRKY6 and WRKY53, transcriptional regulators of SA-defense genes. SBT3.3 promoter region notably increased in pyr1 plants compared to Col-0 plants, while H3K9ac marks remained invariant , for the induction of the ET-mediated triple response. The triple response in Arabidopsis consists of shortening and thickening of hypocotyls and roots and exaggeration of the curvature of apical hooks. Compared to Col-0, the assay revealed that pyr1 seedlings showed no curvature of the apical hook  conferred significantly enhanced resistance. Conversely, in PYR1 loss-of-function mutants, the resistance was compromised. Therefore, the PYR1 receptor had functional specificity in perceiving ABA produced in response to fungal infection to activate plant immunity. This study provided novel information about a specific ABA receptor-mediating specific plant immune responses and pinpointed ABA-activated SnRK2s as cardinal components for plant resistance. This information helps construct a functional classification scheme of the different members of the PYR/PYL receptor family with respect to their downstream signaling pathways in a true biological context. Thus, specific non-redundant roles for PYR1 and PYL8 have been reported in plant immunity (this work) and root ABA sensitivity [PYR1 in vascular bundles , with resistance appearing negatively regulated by ABA, whereas resistance is enhanced in ABA-deficient mutants  [P. syringae DC3000 was severely restricted in pyr1 plants, as documented for aba2 aao3 plants or the ABA-insensitive abi1-1 and abi2-1 mutants [pyr1 mutants, and that pyr1 was insensitive to the damping effect of ABA on SA signaling. This finding supported previous work demonstrating the negative role of ABA on disease resistance to biotrophs, and that P. syringae-induced ABA levels in Col-0 suppress SA biosynthesis and action, enhancing susceptibility to this pathogen [pyr1 plant but ET perception became compromised added a degree of specificity for the understanding of the disease resistance phenotype of the mutant. The observation that in pyr1;NahG plants, the pyr1-mediated ET-insensitivity was reversed, and that SA per se could block ET perception in Col-0 plants (P. syringae DC3000) have developed strategies to alter the host ABA physiology as part of the infection strategy [This study showed that resistance to cumerina ,25,71. Mcumerina , as repocumerina , then AB., 2010) ,21,26,27 mutants . These r mutants ,73,74,75pathogen ,73,75,76strategy ,76. pyr1 plants bear constitutive activation of ICS expression and moderate enhanced level of SA. Besides, pyr1 plants carry the hallmarks of immune priming, including (1) basal activation of MPKs; (2) repression of NRPD2 and, therefore, the RdDM mechanisms that negatively control the onset of defense; (3) activation of the SBT3.3 subtilase; and (4) readying of SA-related genes for enhanced expression by pertinent chromatin modifications. Therefore, pyr1 plants mirror the phenotypes of RdDM defective mutants, which exhibit simultaneous enhanced susceptibility and resistance to necrotrophs and biotrophs, respectively [pyr1 plants. The fact that immune priming and SA-mediated resistance are negatively regulated by the RdDM, and that in pyr1 plants, the ABA repression of SA pathway is relieved, both observations unveil the importance of ABA/PYR1 as new element participating in an epigenetic mechanism of control of gene expression in plant immunity.How does then ABA/PYR1-mediated signaling control SA-mediated defenses? ectively , support−2 s−1 fluorescent illumination) on a 10-h-light and 14-hr-dark cycle. All mutants and transgenic plants are in Col-0 background; SnRK2.6-HA/OE and SnRK2.2-HA/OE were previously described [Arabidopsis thaliana plants were grown in a growth chamber  and purified by lithium chloride precipitation. Reverse transcription was done using the RevertAid H Minus First Strand cDNA Synthesis Kit . Quantitative PCR (qPCR) was performed using an ABI PRISM 7000 sequence detection system and SYBR-Green . HA-tagged SnRK2.2 and SnRK2.6 were immunoprecipitated and used for in vitro kinase assay, as described previously .SBT3.3, PR-1, WRKY6, WRKY53, and Actin2, as reported [Chromatin isolation and immunoprecipitation were performed, as described ,62. Chipreported ,62. All w/v) and transferred to nitrocellulose filters. The filter was stained with Ponceau-S after transfer and used as a loading control.Protein crude extracts were prepared by homogenizing ground frozen leaf material with Tris-buffered saline (TBS) supplemented with 5 mM DTT, protease inhibitor cocktail (Sigma-Aldrich), and protein phosphatase inhibitors . Protein concentration was measured using Bradford reagent; 25 μg of total protein was separated by SDS-PAGE  at 3, 12, and 8 days after inoculation with P. cucumerina and A. brassicicola, respectively. For pathogen-induced callose deposition analyses, infected leaves were stained at 24, 48, and 72 h.p.i. with aniline blue, and callose deposition quantifications were performed, as described by [ollowing ,62. Twelescribed ,24, withribed by .ABA, JA, SA levels were determined, as described previously ,71.


Oleoylethanolamide and palmitoylethanolamide are members of the fatty acid ethanolamide family, also known as acylethanolamides. Their physiological effects, including glucose homeostasis, anti-inflammation, anti-anaphylactic, analgesia, and hypophagia, have been reported. They have affinity for different receptor proteins, including nuclear receptors such as PPARα, channels such as TRPV1, and membrane receptors such as GPR119 and GPR55. In the present review, the pathophysiological functions of fatty acid ethanolamides have been discussed from the perspective of receptor pharmacology and drug discovery. Arachidonylethanolamide , oleoylethanolamide (OEA), and palmitoylethanolamide (PEA) are members of the acylethanolamide or fatty acid ethanolamide family, and are each composed of a fatty acid and an ethanolamine linked together by an amide bond . These a1 and CB2 receptors, along with another ligand, 2-arachidonyl glycerol WIN 55212-2b, the full agonist of CB1/CB2 receptors, by PEA in mast cells expressing CB2 only [2 receptor was also observed in cerebellar granule cells in glutamate-induced neurotoxicity [Aloe et al. reported the pharmacological effects of PEA on mast cells in 1993 . In the CB2 only ,73. PEA toxicity . In additoxicity . Later, toxicity . In additoxicity .2 antagonist, and by OMDM188, a diacylglycerol lipase enzyme inhibitor (DAGL) [2 and GPR55 receptors was observed [Therefore, the acronym ALIA  was proposed for the mechanism of action of PEA in the early 1990s . The actr (DAGL) . PEA sigr (DAGL) . Additioobserved .2, GPR55, and 2-arachidonyl glycerol  prevented the anti-edema and anti-hyperalgesic effects of PEA, suggesting its interaction with a yet uncharacterized CB2-like cannabinoid receptor [The anti-inflammatory action of PEA in a rat model of carrageenan-induced acute hind paw inflammation was compared with that of the non-steroidal anti-inflammatory drug, indomethacin . PEA andreceptor .2 agonists and antagonists suggest involvement of CB2 receptor in pharmacological actions of PEA [1 and CB2 [2 involvement is controversial, multiple targets have been suggested for PEA anti-inflammatory actions, GPR55, TRPV1, and PPARα [N-acyl phosphatidylethanolamine phospholipase D expression and PEA bioavailability, and PEA exerted potent anti-inflammatory actions [1, CB2, and PPARα [2  [Ryberg et al. reported GPR55 as a novel cannabinoid receptor because cannabinoid ligand CP55940 exhibits specific binding to GPR55 . PEA is ectively . Howeverectively ,96,97. Tectively ,99. Phyl2, GPR55, TRPV1, and PPARα [2 antagonists [2 receptor as a target of PEA. However, another putative CB-like receptor was present in the RBL-2H3 cells [2 expression [2 through PPARα activation [1 and CB2) via heteromerization is intriguing. In the co-expression system, CB1 signaling was enhanced by GPR55 expression, and cross-antagonism between GPR55 and CB2 was observed [2 complex is needed.The mechanism of action of PEA has been studied, and many molecular targets have been proposed, including CBnd PPARα . The antagonists ,78,81, tH3 cells . Howeverpression . PEA wastivation . In addiobserved ,102. FurAlthough GPR55 is reported to be a receptor for lipid mediators such as lysophosphatidylinositol and PEA, there have not been many studies on GPR55 being a PEA receptor. Instead, many studies have been conducted using GPR55 knockout mice and the most potent natural ligand, 2-arachidonyl lysophosphatidylinositol, or a synthetic agonist, O-1602 ,104,105.2 or CB2-like receptors has been implicated. However, the likelihood of GPR55 as CB2-like receptor and heterodimeric functions of CB2 and GPR55 in the mast cells have not been studied. Despite being an endogenous ligand for GPR55, the pathophysiological significance of lysophosphatidylinositol in association with GPR55 has poorly been studied in detail. Instead, substantial research work focused on synthetic agonist O-1602 and antagonist CID16020046 in GPR55 knockout mice, which resulted in complex and contradictory reports. Thus, many selective and specific agonists/antagonists need to be developed with respect to GPR55 receptors.Treatment with acylethanolamides OEA and PEA elicit multiple responses through multiple targets. The anti-inflammatory action of PEA may be mediated partially through PPARα. The anti-anaphylactic action of PEA was studied in the early 1990s, and involvement of CBThe anorectic action of OEA seemed to be mediated mainly through PPARα and peripheral vagal sensory nerves. Since the discovery of GPR119 as a receptor for OEA, development of GPR119 agonists as potential pharmacotherapies for diabetes and obesity has been intensively conducted. OEA-induced glucose homeostasis can be mediated by GPR119 receptor, which influences GLP-1 release from L-cells and insulin secretion from β-cells. Drug development targeting GPR119 has been boosted by novel scaffolds such as DS-8500a. However, several synthetic GPR119 agonists induce body weight loss by a GPR119-independent process. In addition, little is known about GPR119 functions in pancreatic α-cells and PP-cells and in PYY secretion from L-cells. Further research is needed in a variety of cell types and organs to elucidate the pathophysiological role of GPR119 in type 2 diabetes and obesity.


Clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated protein 9 (Cas9), a newly developed genome-editing tool, has revolutionized animal and plant genetics by facilitating modification of target genes. This simple, convenient base-editing technology was developed to improve the precision of genome editing. Base editors generate precise point mutations by permanent base conversion at a specific point, with very low levels of insertions and deletions. Different plant base editors have been established by fusing various nucleobase deaminases with Cas9, Cas13, or Cas12a (Cpf1), proteins. Adenine base editors can efficiently convert adenine (A) to guanine (G), whereas cytosine base editors can convert cytosine (C) to thymine (T) in the target region. RNA base editors can induce a base substitution of A to inosine (I) or C to uracil (U). In this review, we describe the precision of base editing systems and their revolutionary applications in plant science; we also discuss the limitations and future perspectives of this approach. Agrobacterium VirD2 relaxase with Cas9 endonuclease and generate potential Cas9-VirD2 for allele modification in rice.The development of genome-editing technology has revolutionized agricultural studies, enabling the improvement of different plant traits ,2,3,4,5.To overcome these limitations, researchers have aimed to develop editing technologies with an improved efficiency, reliability, and precision. Base editing is a newly developed technique for precise genome editing that enables irreversible base conversion at a specific site. The base editor (BE) is a complex of a catalytically impaired Cas protein, guide RNA (gRNA), and a nucleobase deaminase domain which is able to convert specific base pairs ,19,20,21In this review, we discuss the development of various plant BEs (PBEs), focusing on comparing the precision of base-editing tools with that of other CRISPR tools and examining their increasing applications in crop improvement within a short period. Furthermore, future perspectives and current limitations of base-editing tools are discussed.Streptococcus pyogenes (SpCas9), which required a protospacer adjacent motif (PAM) sequence, NGG, was most commonly used. To overcome the G-rich PAM preference, Cpf1  was developed in 2015 for targeting T-rich PAMs [CRISPR/Cas9 was first described in 2002, although the CRISPR mechanism was discovered in 1987 ,32. Cas9ich PAMs ,34. In 2ich PAMs   remain difficult to achieve. Base editing or a specific base conversion enables the replacement of a target base pair with a different base pair in a desirable manner, without inducing DNA DSBs . In contBacillus subtilis bacteriophage PBS1 which blocks the human uracil DNA glycosylase (UDG) activity. The Cas9 nickase (nCas9) and catalytically inactive Cas9 (dCas9) are derived from a wild-type Cas9, with one (nCas9) and two (dCas9) amino acid mutations, respectively. Komor et al. [BEs restrict the generation of indels at both target and off-target sites, as there is no requirement for DNA modification via DSBs ,25,45,46r et al. .Staphylococcus aureus Cas9 (SaCas9) and SaKKH, Qin et al. [Petromyzon marinus cytidine deaminase 1 (PmCDA1) is also efficient as rAPOBEC1 based CBEs [Lachnospiraceae bacterium Cpf1 (dLbCpf1) or Acidaminococcus sp. Cpf1 (dAsCpf1) with rAPOBEC1 and UGI, thus creating dCpf1-BEs, dLbCpf1-BE0 and dAsCpf1-BE0. With further modifications, a series of Cpf1-BEs were developed to improve the editing efficiency.Different types of Cas proteins and engineered Cas9 proteins have been shown to effectively recognize different PAMs and can be utilized as BEs . Severaln et al.  developesed CBEs . Wu et ased CBEs  expandedsed CBEs  reportedsed CBEs ,56. Thissed CBEs . Using tEscherichia coli was mutated to obtain a deoxyadenosine deaminase (TadA*), and TadA-TadA* heterodimer was generated via fusion [ABEs were developed later for converting A to G. Unlike in CBEs, there is no need for a DNA glycosylase inhibitor in ABEs, and natural adenine deaminase cannot accept DNA. Liu’s group developed several generations of ABEs. Transfer RNA adenosine deaminase (TadA) from a fusion . ABEs wea fusion . Hu et aa fusion  replaceda fusion ,60. In pa fusion ,62,63,64Prevotella sp. P5-125 (PspCas13b) was found to be highly specific, and generation of REPAIRv2 via dCas13b–ADAR2DD fusion resulted in a highly specific RNA-editing tool (The CRISPR/Cas effector Cas13a (previously known as C2c2) was engineered for RNA knockdown and binding. An RBE system was developed through the RNA editing for programmable A-to-I replacement (REPAIR) system using a combination of a catalytically inactive Cas13 (dCas13) and an adenosine deaminase acting on RNA (ADAR) . A Cas13ing tool c [15,29,Arabidopsis genomes [In plants, many genes have been edited using various BEs. As a result, precise mutations  have been produced in target regions in the rice, wheat, maize, potato, watermelon, cotton, tomato, and  genomes ,70 (TablSLR1 and NRT1.1B), known to control the plant architecture and effective utilization of nutrients in rice, respectively, were targeted and edited via base editing, which resulted in increases in the height and nitrogen use efficiency in the mutant plants [OsAOS1, OsJAR1, OsJAR2, and OsCOI2), Ren et al. [UGI gene in rBE9 , which can be used for mutagenesis-oriented editing . Successt plants . Moreoven et al. ,73 found in rBE9 .TaALS-P174 gene, with an approximately 75% mutation efficiency, and the mutants showed high tolerance to imidazolinone, sulfonylurea, and aryloxyphenoxy propionate-type herbicides [ALS genes have been shown to confer herbicide resistance to several plant species [TaLOX2 and ALS genes using pnCas9-PBE and A3A-PBE. To date, base-editing systems have been successfully used not only for the modification of genes of herbicide resistance in rice and wheat but also for editing ZmCENH3 and ALS genes in maize and watermelon [ALS gene [StALS and StGBSS genes in potato [ALS gene using a CBE [ALS gene [DELLA and ETR1 genes [GhCLA (responsible for the chlorophyll content) and GhPEBP (controls branching of plants) genes, with mutation efficiencies of 26.67–57.78%. Taken together, these findings demonstrated that base-editing tools might be effectively used for editing of target genes and might have broad applications for the improvement of plant traits.One of the most effective and promising applications of CBEs in plants has been development of herbicide resistance. Recently, an elite herbicide-resistant transgene-free wheat variety was developed by targeting the rbicides . Acetola species ,83. Zong species ,71 also termelon ,70. Resing a CBE . MoreoveALS gene . Marker-R1 genes . In anotR1 genes . Base edACC, ALS, CDC48, DEP1, NRT1.1B, and OsEV) were edited using a successfully developed ABE [IPA1 (OsSPL14), OsSPL17, OsSPL18, and SLR1 genes. Two ABEs, ABE7.8 and ABE7.10, were used to edit MPK6, MPK13, SERK2, WRKY45, and Tms9-1 genes [OsACC, OsALS, OsDEP1, OsNRT1, OsCDC48, and OsWx genes in rice [DEP1, TaEPSPS, and GW2 genes [Arabidopsis genes  and two rapeseed genes (BnALS and BnPDS) were edited with the same strategy [AtFT-knockout transgenic plants were found to exhibit delayed flowering, whereas AtPDS3-knockout transgenic plants were found to exhibit dwarfism and a mosaic albino phenotype using ABE7.10 in Arabidopsis [Notable findings were reported in studies developing specific traits in plants using evolved BEs (from useful BEs to effective PBEs), and the efficiencies of these BEs were also evaluated. Li et al.  reportedoped ABE . Furtheroped ABE  evaluate-1 genes ,76, sugg in rice . Similar in rice ,47,85. F in rice . Liang e in rice  develope in rice . Using AW2 genes . Four Arstrategy . As a rebidopsis . Thus, bAlthough base-editing systems, with their simplicity and precision, have rapidly advanced genome editing, there are still many aspects of this technology that should be explored  and manyThe second challenge is off-target mutations, which are considered a major limitation of genome editing systems. Off-target activity of BEs or Cas9-linked deaminases is still not clear. Kim et al.  reportedArabidopsis agronomic traits, such as flowering time, plant height, pathogen resistance and albino phenotype by editing AtFT, AtPDS3, and AteIF4E genes [Furthermore, ABEs can generate efficient mutations and alter 4E genes ,81. BEs 4E genes . Both AB4E genes ,53,69,814E genes . However4E genes  recently4E genes , and theFinally, BEs can target either C or A within the editing window; however, it remains unknown which base will be converted within an editing window. Generally, BEs with larger editing windows, such as A3A-PBE, are desirable because of a large number of editable bases . HoweverBase editing is a recently developed and advanced technology. However, there is still a need for developing easier, more precise, and more convenient base-editing tools. Base-editing tools have been developed within a shorter period of time than were other CRISPR-based tools. Thus, more applications of base-editing systems in crop improvement are expected in the near future.


In recent years, impurity-doped nanocrystal light-emitting diodes (LEDs) have aroused both academic and industrial interest since they are highly promising to satisfy the increasing demand of display, lighting, and signaling technologies. Compared with undoped counterparts, impurity-doped nanocrystal LEDs have been demonstrated to possess many extraordinary characteristics including enhanced efficiency, increased luminance, reduced voltage, and prolonged stability. In this review, recent state-of-the-art concepts to achieve high-performance impurity-doped nanocrystal LEDs are summarized. Firstly, the fundamental concepts of impurity-doped nanocrystal LEDs are presented. Then, the strategies to enhance the performance of impurity-doped nanocrystal LEDs via both material design and device engineering are introduced. In particular, the emergence of three types of impurity-doped nanocrystal LEDs is comprehensively highlighted, namely impurity-doped colloidal quantum dot LEDs, impurity-doped perovskite LEDs, and impurity-doped colloidal quantum well LEDs. At last, the challenges and the opportunities to further improve the performance of impurity-doped nanocrystal LEDs are described.  As a result, the optimized FA(1−x)CsxPbBr3 PeLED exhibited the maximum luminance and CE of 55,005 cd m−2 and 10.09 cd A−1, respectively, suggesting 6.4- and 3.7-fold higher than FAPbBr3 PeLEDs. In particular, the luminance of 55,005 cd m−2 was the highest for nanocrystal PeLEDs at that time, which resulted from the proper energy level, homogeneous film morphology, and the improved stability of perovskites. Sun et al. took the first step to develop green PeLEDs by utilizing mixed-cation perovskite emitters, where cations in FA doping) . Two maj+ doping has a great influence on the crystal growth, structure, photoelectric, and optical characteristics of FAPbBr3, which importantly improved the PLQY of FAPbBr3 film (~10-fold) on account of the substantially suppressed trap density [3 were improved by ∼10-fold and ∼5-fold to 66,353 cd m−2 and 24.22 cd A−1 compared to the controlled devices, respectively, which were the highest for FAPbBr3-based PeLEDs at that time. On the other hand, the realization of Cs+ doping in MA-based perovskites [+ doping in FA-based perovskites [+ doping in Cs-based perovskites [Later, Wu et al. found that Rb density . Hence, ovskites , MA+ dopovskites , and FA+ovskites  have als2+ with isovalent or heterovalent cations via well-designed synthetic methods is also effective in enhancing the device performance, which is similar to the situation in impurity-doped red PeLEDs. For example, the maximum EQE was increased from 0.81% for pure CsPbBr3 PeLEDs to 1.49% for Mn2+-doped CsPbBr3 PeLEDs [3 PeLEDs to 4.4% for Ce3+-doped devices [−2 for pure CsPbBr3 PeLEDs to 12,500 cd m−2 for Sn4+-doped PeLEDs [2+-doped CsPbBr3 PeLEDs was up to 13.13 cd A−1, which was a ~100-fold improvement compared to undoped counterparts [2+ doping in CsPbBr3 is easier relative to many other divalent ions  since CsSnBr3 and CsPbBr3 possess similar ABX3-type perovskite crystalline structures [2+-based CsSnX3 is unstable since Sn2+ easily oxidizes to Sn4+, resulting in low PLQY [4+ doping in CsPbBr3 is more approximate than Sn2+ doping for PeLEDs [In the case of B-site doping for green PeLEDs, the partial replacement of Pb3 PeLEDs , while t devices . Additiod PeLEDs . Remarkaterparts . In partructures ,302,303.low PLQY . Hence, r PeLEDs .2+ doping is widely adopted to enhance the performance of blue all-inorganic perovskites [3 and CsMnX3, (ii) higher formation energies of CsMnX3 than those of CsPbX3 to avoid the thermal instability issue that is associated with the intrinsically low formation energies of perovskite lattices, iii) smaller ion radius of Mn2+ (∼0.97 Å) [Compared with impurity-doped red and green PeLEDs, relatively little attention has been paid to impurity-doped blue PeLEDs. This may be because it is more difficult to synthesize high-performance blue perovskites together with the fact that it becomes harder to manipulate the device engineering due to the wide bandgap of blue emitters ,307,308.ovskites ,311. The∼0.97 Å) ,313,314.2+ increased the PLQY over three-fold for CsPbCl3 films (28%) without an Mn-emission peak [2+ for Mn emissions. By employing a two-step synthetic scheme to adjust Mn2+ doping, the PLQY and lifetime were increased while trap states were reduced. In addition, perovskites became more monodisperse, narrowing the emission bandwidth. PeLEDs were constructed with the device architecture of ITO/PEDOT:PSS/TFB/PFI/perovskites/TPBi/LiF/Al, where Mn-doped CsPbCl3 was blue perovskite emitters, as shown in 2+ saturated the emission in the thin EML layer and TFB may further lower Mn emission, enabling the mild doping for enhanced performance without sacrificing color purity. Compared with undoped PeLEDs, the maximum of EQE of PeLEDs with the 0.19% Mn doping showed a four-fold improvement, reaching 2.12%, which was the highest for blue PeLEDs at that time [2+ doping could enhance the luminance, efficiency, and stability of bulk sky-blue CsPbBr1.9Cl1.1 PeLEDs [−2 was yielded, which was among the highest for blue PeLEDs. More recently, Song et al. reported that the maximum EQE of Ni2+-doped blue CsPbBr0.99Cl2.01 PeLEDs was up to 2.4%, which was the best for blue CsPbX3 PeLEDs [The first successful Mn-doped blue PeLEDs was reported by Congreve et al., where a small amount of Mnion peak . A signihat time . Later, 1 PeLEDs . In this3 PeLEDs . −1 [−1 [Since the first demonstration of CQW-LEDs in 2014 , the inv−1 , which i−1 [−1 ,324,325.−1 [−1 ,327,328.−1 [−1 . Further3/Al, where CQWs with different doping concentrations served as the emitters, CBP and TAPC HTLs were used to understand the device engineering. Firstly, CQW-LEDs with 0% Cu-doped concentration exhibited the narrow EL FWHM of 12 nm and the Commission Internationale de L’Eclairage (CIE) 1931 coordinates of , rendering than the color gamut covered 104% of the International Telecommunication Union Recommendation BT 2020 (Rec. 2020) standard in the CIE 1931 color space. Secondly, CQW-LEDs with 0.5% Cu-doped concentration possessed dual emission with an EQE of 0.146% (+ emission or holes at CdSe VBM for CdSe emission). In the case of CBP-based CQW-LEDs, the Cu+ emission was lowered with increasing voltage. In greater detail, the charge trapping issue existed in the doped LEDs, since holes transported from CBP were more easily trapped by Cu under a low electrical field while saturated at high voltages due to the high Cu level (5.05 eV) compared with CdSe VBM (5.69 eV), resulting in relatively more holes transported from CBP being injected into the VBM of CdSe after saturation at the dopant site. Hence, a lower current density in doped LEDs was obtained as compared to undoped counterparts. Furthermore, the ideality factor for the doped LEDs was nearly twice that of undoped counterparts, suggesting that Cu-doping was an impurity defect site.In terms of impurity-doped CQW-LEDs, only one work has been reported to date . Specifif 0.146% , demonst+, since holes were readily injected into Cu due to the barrier-free characteristic between the HOMO of TAPC (5.4 eV) and Cu level while the existing hole barrier between TAPC and CdSe. Finally, a white LED based on CQWs was explored, in which a high Cu-doped concentration of 2.4% was used. Such findings could be further extended to other impurity -doped CQWs to realize LEDs, considering the well-developed impurity-doped CQD-LEDs and PeLEDs. Therefore, the factor of impurity-doped materials played a significant role in improving the efficiency and stability, while the factor of device architecture affected the emission mechanism in impurity-doped CQW-LEDs.For TAPC-based CQW-LEDs, the EL emission peak of CdSe was lower than that of CuBy virtue of impurity doping, the electronic, optical, catalytic, transporting and magnetic properties of nanocrystals can be controlled to satisfy the requirement of optoelectronic and microelectronic applications. With the gradual comprehending of the effect of impurity doping , the development of impurity-doped nanocrystal LEDs is flourishing ,330,331.After extensive efforts made by researchers worldwide, the performance of impurity-doped nanocrystal LEDs has been gradually improved. Given the facile solution-processed fabrication procedures, it is believed that impurity-doped nanocrystal LEDs can be well applied to low-cost flexible electronics and transparent products ,333,334.Currently, some effects of impurity-doped nanocrystal LEDs are still unknown. For example, (i) only a few impurity-doped blue nanocrystal LEDs have been studied, limiting the general full-color applications; (ii) in spite of the remarkable evolution of impurity-doped nanocrystal emitters, the deep insight of device engineering is urgently required to be explored; (iii) although impurity-doped visible-color nanocrystal LEDs have been widely investigated, scarce attention is paid to infrared and ultraviolet devices; (iv) despite rare earth impurity-doped nanocrystals having been intensively probed, almost no LEDs based on this type of emitter have been reported; (v) pursuing the real commercialization of impurity-doped nanocrystal LEDs still faces a number of challenging tasks, including efficiency, efficiency droop, toxicity and lifetime.For the traditional III-Nitride-based LEDs, the maximum EQE exceeds 84% , while h−2 for displays and ≥10,000 h at ≥1000 cd m−2 for the solid-state lighting are necessary) [2O3 and SiO2 atomic layer deposition [−6 g−1 m−2 day−1) [To overcome the toxicity problem, more endeavors should be taken in the development of heavy-metal-free impurity-doped nanocrystal LEDs ,354,355,cessary) ,358,359.cessary) ,362,363 position  and orgaposition  to reduc2 day−1) ). Upon l2 day−1) ,371,372.