Uncoupling proteins (UCPs) are transmembrane proteins members of the mitochondrial anion transporter family present in the mitochondrial inner membrane. Currently, six homologs have been identified (UCP1-6) in mammals, with ubiquitous tissue distribution and multiple physiological functions. UCPs are regulators of key events for cellular bioenergetic metabolism, such as membrane potential, metabolic efficiency, and energy dissipation also functioning as pivotal modulators of ROS production and general cellular redox state. UCPs can act as proton channels, leading to proton re-entry the mitochondrial matrix from the intermembrane space and thus collapsing the proton gradient and decreasing the membrane potential. Each homolog exhibits its specific functions, from thermogenesis to regulation of ROS production. The expression and function of UCPs are intimately linked to diabesity, with their dysregulation/dysfunction not only associated to diabesity onset, but also by exacerbating oxidative stress-related damage. Male infertility is one of the most overlooked diabesity-related comorbidities, where high oxidative stress takes a major role. In this review, we discuss in detail the expression and function of the different UCP homologs. In addition, the role of UCPs as key regulators of ROS production and redox homeostasis, as well as their influence on the pathophysiology of diabesity and potential role on diabesity-induced male infertility is debated. It is currently estimated that 9.3% of the world population suffers from diabetes mellitus, whereas 39% of adults aged 18 years and over are overweight, and 13% are obese . Obesity by 70%) . Proton  by 70%) . On the  by 70%) .Herein we discuss in detail the expression and function of the different UCP homologs in human, mouse, and rat. In addition, the role of UCPs as key regulators of ROS production and redox homeostasis and their potential role on the pathophysiology of diabesity and on diabesity-induced male infertility will be discussed.UCPs are integral transmembrane proteins of the inner mitochondrial membrane and members of the mitochondrial anionic transporter family SLC25 ,9,11,12.UCPs play a central role in regulating the potential of mitochondrial membrane, exhibiting several distinct functions from thermogenesis (dissipating energy in the form of heat) to oxidative phosphorylation or ROS levels regulation ,18,20. CUCP1 mRNA in human ductus deferens, testis, epididymis, seminal vesicle, and prostate [UCP1 is predominantly expressed in brown adipose tissue (BAT), where it constitutes up to 10% of the mitochondrial proteins in this tissue ,79. Humaprostate  has beenUcp1 [Ucp1 mRNA levels drop with starvation and recover after feeding [The main function attributed to UCP1 is the production of heat through thermogenesis . BAT maiUcp1 . As a reUcp1 . These fUcp1 . In addiUcp1 , UCP1 acUcp1 , and comUcp1 . In addi feeding . UCP1 is feeding . In contUcp2 mRNA was reported in pancreatic β-cells [UCP2 mRNA has been predicted in human ductus deferens, testis, epididymis, seminal vesicle, and prostate [UCP2, which is 59% identical to UCP1, is probably the most widely expressed UCP homolog ,93. Unli β-cells , skin [3 β-cells , female  β-cells , brain [ β-cells , WAT, BA β-cells , and liv β-cells . In rats β-cells , neurons β-cells , microgl β-cells , heart [ β-cells , testis  β-cells , and pan β-cells , and its β-cells , skeleta β-cells , WAT, BA β-cells , lungs [ β-cells , spleen  β-cells , and thy β-cells . Additioprostate .Ucp2 mRNA expression is reported to be regulated by glutamine and leptin [The main function attributed to UCP2 comprises the regulation of ROS production and protection against oxidative damage ,95. Thusd leptin ,109. On d leptin ,107.UCP3 mRNA was predicted in human ductus deferens, testis, epididymis, seminal vesicle, and prostate [UCP3 shares an 73% identity with UCP2 and 57% with UCP1 ,60. In hprostate .UCP3 mRNA is also regulated by fasting in both human and rat [Similar to UCP2, UCP3 also plays a role on the regulation of ROS production and protection against oxidative damage ,111,112. and rat ,117.UCP4 mRNA in human ductus deferens, testis, epididymis, seminal vesicle, and prostate [UCP4, along with UCP5 and UCP6, are less identical to the other three homologs . UCP4 isprostate  has been2+ signalling [Ucp4 mRNA expression in the rat brain as well [The activation of UCP4 attenuates mitochondrial ROS levels . Since tgnalling . Analogognalling . Calorie as well .UCP5 mRNA has been predicted in human ductus deferens, epididymis, seminal vesicle, and prostate [UCP5 is mainly present in human , rat 7575 and moprostate .Ucp5 mRNA levels decrease during brain hypoxia and increase under hyperoxic conditions [2S, so UCP5 is a potential regulator of the mitochondrial levels of this important signalling molecule. UCP5 also efficiently transports other inorganic anions, including sulphite, phosphate and, to a lesser extent, maleate, oxalate, malonate, malate, citramalate, aspartate, and glutamate [As UCP4, UCP5 may have a neuroprotective effect against damage related to oxidative stress . This canditions . UCP5 isnditions . Interesnditions . These clutamate .Ucp6 mRNA was found to be mainly expressed in mouse kidneys [UCP6 mRNA was predicted in human ductus deferens, testis, epididymis, seminal vesicle, and prostate [ kidneys . It is aprostate .2S concentration, which at low concentrations inhibits ROS production in mitochondria [Ucp6 was found to be upregulated during cancer [Ucp6 expression is associated with increased mitochondrial activity [2S levels [UCP6 is upregulated in response to oxidative stress . Althougchondria . In factg cancer  or the rg cancer , which aactivity . SimilarS levels . UCP6 isS levels .2 at the oxidative chain level, which results in formation of the superoxide anion. However, an unregulated increase in the production of these free oxygen radicals renders the cellular antioxidant system incapable of mitigating their harmful effects, causing a shift in the homeostasis of the redox balance towards oxidative stress, a damaging state that participates on the pathophysiology of several diseases and capable of inducing macromolecular damage towards apoptosis/necrosis [ROS are an essential part of the cell function, participating in numerous processes such as immune response, cellular proliferation and differentiation, angiogenesis, aging, and several signaling pathways. They are a natural byproduct of the mitochondria’s oxidative phosphorylation, arising when a single unpaired electron reduces Onecrosis ,126. It necrosis ,128,129.Ucp1 knockout mice exhibited greater H2O2 production and higher mitochondrial membrane potential [Ucp1 in the BAT mitochondria of mice exposed to cold stress leads to higher superoxide production, and subsequently oxidative stress, when compared to wild-type mice [Ucp1 deletion in mice does not predispose them to higher oxidative damage [Ucp1 knockout mice, they determined that UCP1 activity bears no weight in the reduction of ROS emission, except when succinate was endogenously added. Interestingly, the authors also inferred that low membrane potential may not necessarily correlate with lower oxidative stress, challenging the claim that mild uncoupling protects mitochondria against oxidative damage [All UCPs homologs are well known for regulating ROS homeostasis, although UCP1 role as an antioxidant remains debatable. Given the extensively described uncoupling activity of UCP1, it would be expected that this protein has an active role in the control of mitochondrial ROS production. However, there are some conflicting data. For instance, Dlasková et al. observed that BAT mitochondria from both wild-type C57BL/6J mice treated with guanosine diphosphate (GDP), an UCP1 inhibitor, and otential . In accoype mice ,132. Besype mice . Converse damage . In a lae damage .2O2 generation, whereas the treatment in mitochondria with low expression of UCP2 led to no changes in the oxidative state [Ucp2 in multiple mice strains induces chronic high levels of ROS in pancreatic cells [Ucp2-deficient mice also results in higher levels of oxidative stress markers in the liver and delayed hepatic regeneration [Ucp2 knockout mice infected with Toxoplasma gondii presented a stronger infection fighting capacity, when compared to wild type, due to higher ROS emission in macrophages [UCP2 and UCP3 role as modulators of ROS production is extensively described. UCP2 and UCP3 do not display a clear uncoupling activity, so it is unlikely that they regulate superoxide production in a mitochondrial uncoupling-dependent manner. The redox balance mediated by UCP2 was first demonstrated by Nègre-Salvayre et al., where it was shown that mitochondria fractions containing high levels of UCP2 treated with the inhibitor GDP presented a significant increase in Hve state . Other sve state ,97. UCP2ve state ,137,138.ic cells ,140,141.neration , as wellneration . On anotrophages ,144,145.Ucp3 knockout mice display higher rates of ROS production and are more vulnerable to oxidative damage, including increased vulnerability to ischemia-reperfusion injury [Ucp3 knockout mice and reported that UCP3 decreases the rate of ROS production [Akin to UCP2, it was hypothesized that the overproduction of ROS activates UCP3 through a negative feedback mechanism, diminishing oxidative damage . The pren injury ,151,152.oduction . UCP3 isoduction ,154. Oveoduction . On a fioduction .Ucp4 and Ucp5 mRNA and respective proteins were found to protect neurons against oxidative stress damage [2S levels. Although H2S exerts cytotoxic effects when in high concentrations, at low concentrations it exhibits beneficial and protective effects [2S concentrations is the inhibition of ROS production in mitochondria through the stimulation of cellular antioxidants [2S low.Although fewer studies focus on UCP4-6, compelling evidence highlights their role on the regulation of ROS production. Increased levels of s damage . In addis damage , whereass damage . UCP6 res damage . It was  effects . One of oxidants ,123. HenUCPs have a significant influence in mitochondrial bioenergetics, thermogenesis, and control of ROS production, which are all key processes known to progressively become impaired with the onset of metabolic disorders (including obesity and diabetes mellitus). Obesity and diabetes mellitus are closely linked, where a prolonged and persistent positive intake of energy lead to chronic hyperglycemia and subsequent insulin resistance . Even thUcp1 knockout mice were incapable of establish a clear connection between UCP1 deficiency and obesity. Enerbäck et al. were the first to report the absence of an obesogenic effect, as Ucp1-deficient mice fed a standard or high-fat diet did not become obese [Ucp1 knockout mice were housed at thermoneutral temperature , they do indeed develop obesity even when fed standard diet. In accordance with these results, more recent studies have also described the obesogenic role of Ucp1 ablation in mice kept at ~30 °C [Ucp1 knockout mice fed Western diet and kept at 25 °C developed insulin tolerance, although no significant differences in body weight, visceral adiposity, and energy expenditure in comparison with wild-type mice were observed [Ucp1 also causes whitening of BAT [Ucp1-deficient mice lack BAT glucose uptake after norepinephrine administration, suggesting that sympathetic stimulation leads to UCP1 mediated glucose disposal by BAT [Ucp1 knockout mice was able to eliminate glucose intolerance [UCP1-mediated thermogenesis can increase 12–20% the total daily energy expenditure . Howeverme obese . These fme obese ,165. Notme obese . Mice hot ~30 °C ,168, evet ~30 °C . Interest ~30 °C . In addiobserved . Loss ofg of BAT , decreasg of BAT ,173. In l by BAT . Notablyolerance . Furtherolerance ,176. In olerance . Furtherolerance ,179. Morolerance ,183,184.Ucp2 knockout mice had improved glucose-stimulated insulin secretion (GSIS), indicating that UCP2 activity negatively regulates insulin secretion [Ucp2 had enhanced β-cell responsiveness to glucose, decreased levels of glycemia and restored first phase insulin secretion. Overall, these results seem to indicate that obesity-dependent induction of UCP2 in pancreatic β-cells has a pathogenic effect towards the development of insulin resistance and in turn diabetes mellitus. In accordance with this hypothesis, overexpression of UCP2 in isolated β-cells was reported to impair GSIS [Ucp2-ablated mice . In addiewed in ,186,188.is model . ATP is is model ,187. Grois model ,192,193. MnTMPyP . A studysed GSIS . Interesotential . Taken tUcp3−/− mice failed to demonstrate an obesogenic effect for Ucp3 ablation [Ucp3 deficiency with protection from diet-induced obesity [Ucp3 ablation have conflicting evidence as Ucp3−/− animals fed a fatty diet exhibited either enhanced [UCP3 mRNA and protein in prediabetic and diabetic individuals, when compared to healthy individuals [UCP3 plays an active role in ROS production and fatty acid oxidation, which makes it a candidate for better understanding the pathophysiology of metabolic disorders such as obesity and type 2 diabetes mellitus. Experimental evidence from multiple studies have proposed UCP3 as a modulator of energy metabolism and a possible protective role in obesity. For instance, transgenic mice overexpressing UCP3 fed a fatty diet exhibit lower body weight, decreased adipose tissue, and triglyceride accumulation ,196,197.ablation ,199. Mor obesity . However obesity ,202. Stuenhanced , decreasenhanced  or unaltenhanced  insulin enhanced . Concurrividuals ,205,206.2O2 production and loss of spermatozoa mobility was observed [The link between oxidative stress and male factor infertility is well documented since it was first described by MacLeod in 1943, where a connection between Hobserved . In factobserved ,212,213.observed . Severalobserved ,218,219.Interestingly, ROS have been described as a double-edged sword for spermatozoa. ROS have a crucial influence in sperm physiology and spermatogenesis, as well as in the fertilization process. Seminal ROS participate as a secondary messenger in several pathways necessary for spermatozoa hyperactivation, capacitation, sperm–oocyte fusion, chemotaxis, acrosome reaction, and sperm head decondensation ,222,223.As previously mentioned, individuals who suffer from obesity or diabetes mellitus present augmented levels of ROS, which results in oxidative stress and ultimately subfertility or even infertility. In fact, the deleterious effects of diabetes mellitus and obesity in male reproductive function are well established. Patients suffering from diabetes and/or obesity exhibit poor sperm parameter, decreased motility, abnormal morphology, reduced density, and higher damage to both nuclear and mitochondrial sperm DNA ,237. It Ucp2 mRNA expression was increased in the mouse testis of experimental cryptorchidism. Hyperthermia in the testis arises from multiple pathological conditions such as varicocele, cryptorchidism, or obesity, and correlates with exacerbated spermatozoa apoptosis [2O2 was shown to induce UCP2 expression [Further understanding of the nefarious impact of ROS production and antioxidant mechanism in male related infertility will surely lead to more focused treatment and accurate diagnostic, as well as better outcomes for subfertile and infertile men. Hence, UCPs by virtue of their antioxidant activity may represent a clinically relevant target to improve reproductive outcomes. UCPs seem to play an important part in mediating the deleterious effects of diabesity in male infertility and subfertility, however little is known about their function in the male reproductive tract. Given the extensively described role of UCPs as key regulators of the mitochondria redox state, it should not come as a surprise that most of the experimental data comes from studies on the activity of UCPs in the testis/sperm ROS production, more precisely UCP2. Zhang et al. reported that UCP2 protects germ cells from hyperthermia-derived ROS production. Furthermore, poptosis . In anotpoptosis . Moreovepoptosis . In humapoptosis . The linpoptosis . Additiopression . AlthougUCPs are widely expressed throughout the organism and play an important role on ROS production and homeostasis. UCP1 is responsible for thermogenesis, but the other homologs display broader functions, including insulin release and regulation of oxidative phosphorylation. Due to their functions, UCPs take part in the pathophysiology of diabesity, where its altered expression and/or dysfunction is associated with the onset of the disease. Studying UCPs expression and function will lead to a better understanding of the pathophysiology of metabolic diseases and potentially to novel therapeutic targets. As mentioned above, diabesity is positively associated with high oxidative stress and, consequently, to male subfertility or even infertility, which is one of the most overlooked comorbidities. Working towards a better understanding of the relationship of UCPs activity with male reproductive function could lead to the development of novel fertility therapies and better reproductive outcomes. However, not much is known concerning the expression and function of UCPs in the male reproductive tract. Further studies are needed to disclosure the role of UCPs in male fertility and redox signaling and homeostasis during spermatogenesis and spermatozoa physiology.


Cellular miRNAs are expressed in tissue fluids with sufficient amounts and were identified as potential molecular targets for studying the physiological mechanisms and correlations with many human diseases particularly diabetes. However, molecular-based changes among older adults with diabetes mellitus (DM) are rarely fully elucidated.  This study is aimed at identifying circulating miRNAs, which hold the potential to serve as biomarkers for the immune-inflammatory changes in older T2D patients with moderate and poor glycemic control status. In addition, the association of both myokines and osteopontin (OPN) levels with circulating miRNAs was identified. α, and CRP), osteopontin, and myokines (adropin and irisin) were estimated by colorimetric and immune ELISA assays, respectively. In addition, real-time RT-PCR analysis was performed to evaluate the expression of circulating miRNAs, miR-146a and miR-144, in the serum of all diabetic subjects.  A total of 80 subjects aged 20–80 years were invited during the period of October 2017–May 2018 to participate in this descriptive cross-sectional study. All subjects were diagnosed with T2D for more than 5 years. Subjects were grouped based on glycemic control  into two groups: moderate glycemic control  and poor glycemic control , respectively. Diabetic control parameters, fasting blood sugar (FS), HbA1c, fasting insulin (IF), insulin resistance (IR), HOMA-IR, inflammatory cytokines (IL-6, IL-8, IL-18, IL-23, TNF-α, CRP, and OPN and a reduction in the levels of myokines, adropin and irisin, compared to patients with moderate glycemic control. The results obtained are significantly correlated with the severity of diabetes measured by HbA1c, FS, IF, and HOMA-IR. In addition, baseline expression of miR-146a is significantly reduced and miR-144 is significantly increased in T2D patients with poor glycemic control compared to those with moderate glycemic control. In all diabetic groups, the expression of miR-146a and miR-144 is significantly correlated with diabetic controls, inflammatory cytokines, myokines, and serum levels of OPN. Respective of gender, women with T2D showed more significant change in the expressed miRNAs, inflammatory cytokines, OPN, and serum myokine markers compared to men. ROC analysis identified AUC cutoff values of miR-146a, miR-144, adropin, irisin, and OPN expression levels with considerable specificity and sensitivity which recommends the potential use of adropin, irisin, and OPN as diagnostic biomarkers for diabetes with varying glycemic control status.  In this study, T2D patients with poor glycemic control showed a significant increase in the serum levels of IL-6, IL-8, IL-18, IL-23, TNF- In this study, molecular expression of certain microRNA species, such as miR-146a and miR-144, was identified and significantly associated with parameters of disease severity, HbA1c, inflammatory cytokines, myokines, and serum osteopontin in T2D patients with moderate and poor glycemic control. The AUC cutoff values of circulating miRNAs, miR-146a and miR-144; myokines, adropin and irisin; and serum OPN were significantly identified by ROC analysis which additionally recommends the potential use of these biomarkers, miR-146a, miR-144, adropin, irisin, and OPN, as diagnostic biomarkers with considerable specificity and sensitivity for diabetes in patients with varying glycemic control status. Diabetes mellitus is the most drastic disease that prevailed among older adults . More thType 2 diabetes mellitus (T2DM) is a progressive disease significantly associated with clinically impaired glucose regulation . The proPhysiological changes in the cellular proinflammatory cytokines were reported in patients with diabetes which significantly play a critical role in the pathogenesis of diabetic complications via multiple biochemical and cellular pathways –13.α (TNF-α), IL-6, IL-18, and IL-1β were reported among patients with type 2 diabetes mellitus displaying features of the insulin resistance syndrome [The development of diabetes and glucose disorders has epidemiologically predicted the change in inflammatory markers , 15, whesyndrome .Also, other cytokines such as osteopontin (OPN), a multifunctional protein secreted from several different cell types including bone cells and adipocytes, showed to be involved in the regulation of human glucose metabolism. Any alterations of these cytokines might be convoluted in the pathogenesis of T2DM. In patients with T2DM, OPN has been shown to be increased probably involved in subclinical inflammation  and insuHigher serum levels of OPN are significantly correlated with diabetic complications such as severe diabetic albuminuria and glomerulosclerosis various models of diabetic nephropathy , 25–27, The correlation of cellular myokine secretion and the severity of diabetes were identified in subjects with DM type 2 , 31. Preβ-cell function, and disease complications of diabetes [Recent molecular-based studies recommended cellular miRNAs as potential targets for studying the physiological mechanisms and correlations with many human diseases –42. Circdiabetes –46. The diabetes –49.Although circulating miRNA levels were assessed recently in younger patients with T1D and correlated positively with the variations in cytokine levels as measures of immune-mediated signaling pathways proceed in the pathogenesis of diabetes –52, littA total of 80 subjects aged 20–80 years were invited during the period of October 2017–May 2018 to participate in this descriptive cross-sectional study. Based on the American Diabetes Association criteria , the subA colorimetric assay was performed to estimate blood glucose for each participant using the QuantiChrom Glucose bioassay kit . In addition, HbA1c and insulin serum levels were estimated using a commercial kit  for HbA1c and immune assay ELISA kit  for insulin, respectively. Insulin resistance in the fasting state was determined from the data of fasting insulin (IF) and fasting glucose (GF) using a validated homeostasis model assessment (HOMA-IR) as previously reported –57.A quantitative immunosorbent assay (ELISA) technique was performed to estimate the concentrations of OPN in the serum of each subject by using a human osteoprotegerin (OPG) ELISA kit (RayBio® (OPG) ELISA Kit, catalogue #: ELH-OPG, Norcross, Georgia 30092). The procedures proceed according to the prescribed manufacturer's instructions, and the concentration of OPN was measured at 450 nm immediately by a complete set of ELISA reader model SLT Spectra 216687.α, and CRP by using a Quantikine Human Immunoassay ELISA kit . The procedures were run according to an accurate ELISA manufacturer's protocol. A standard curve was used to determine cytokine levels, and the concentration of each cytokine was expressed as pg/mL.An enzyme-linked immunosorbent assay was performed to estimate the levels of IL-6, IL-8, IL-18, IL-23, TNF-μg/mL for irisin with interassay coefficients of variance < 10% and <12%, respectively. In addition, the detection range of adropin was 5-1000 ng/L and for irisin was 0.05-15 μg/mL [Serum adropin and irisin levels were estimated by immune assay technique as previously reported  by using15 μg/mL .For each participant, the miRNeasy isolation kit  was used to extract total RNA from serum samples. A reverse transcription polymerase chain reaction (RT-PCR) was applied to analyze total RNA in all serum samples. Then, a complementary DNA (cDNA) was generated using reverse transcription miScriptII RT kits (Qiagen), and the levels of miRNAs were evaluated by optical density , 5.The primers of circulating miRNAs, miR-146a and miR-144 , were used to screen the expression of miRNAs in the plasma of all participants by using quantitative real-time RT-PCR . The avePower calculations of the selected sample size of 80 subjects showed to give an estimated power of 95% and a significance level of 0.05 with an expected frequency of 11.6%.χ2 test were used to analyze the frequency of the differences between the studied groups, respectively.An SPSS statistical program  was used to analyze all data produced in this study. The data of continuous variables are expressed as the mean ± SD. A nonparametric test (Mann-Whitney-Wilcoxon test) and the t-tests were used for comparison between the studied variables such as diabetes (dependent variable) and expression levels of miRNAs, cytokines, myokines, and serum osteopontin (independent variables). In addition, multiple stepwise regressions and Pearson's correlation analysis were used to estimate the associations between diabetes and the studied independent variables in patients with type 2 diabetes. The correlation coefficient was translated into descriptors like “weak,” “moderate,” or “strong” relationship. It is mostly agreed that a coefficient of <0.1 indicates a negligible and >0.9 a very strong relationship, whereas values inbetween are differential [p < 0.05.Two independent sample erential , 61. Theerential . All tesIn this study, the analysis was performed on a total of 80 serum specimens . Based on HbA1c controls, the subjects were grouped into two groups: moderate glycemic controlled  and poor glycemic controlled  type 2 diabetes (T2D) subjects, respectively . The resβ, and TNF-α were evaluated compared to moderate glycemic controlled T2D patients (p = <0.01). In addition, serum CRP levels significantly increased (p = <0.001) in poorly controlled T2D compared to moderately controlled T2D patients (HbA1c > 7-8%)  in all studied cytokines compared to males of the same group.In this study, in order to explore the potential role of the parameters of glucose control in cellular inflammation, the levels of proinflammatory cytokines and myokines were estimated and compared based on diabetes status . In indi > 7-8%) . Additiopatients . Accordipatients  and poorpatients  groups sp = 0.001) reduced in patients with poorly controlled T2D compared with moderately controlled T2D patients (p = 0.001) in the studied myokines, irisin and adropin, respectively, compared to male subjects of the same group as shown in Figures Interestingly, participants with poorly controlled T2D tended to have lower levels of irisin and adropin compared to moderately controlled T2D patients. Both serum levels of adropin and irisin were significantly  in the serum of poor glycemic controlled patients compared to moderate glycemic controlled patients. The levels of miR-146a significantly decreased, and the levels of miR-144 significantly increased in poor glycemic controlled patients compared to moderate glycemic controlled patients  in patients with poor glycemic control compared to that present in patients of moderately controlled HbA1c levels which showed moderate rates of correlation strengths , and HOMA-IR levels, respectively . Both mitrengths .In addition, ROC analysis was performed to explore the potential use of miR-146a, miR-144, OPN, adropin, and irisin expression levels as diagnostic biomarkers for both moderate and poor glycemic controlled patients as shown in Tables Similarly, in T2D patients with poor glycemic control, ROC analysis showed variable values of the studied biomarkers and their correlation with the severity of the T2D. The AUC analysis was 0.91 (0.72-0.98) for miRNA-146a, with a sensitivity of 94.5% and specificity of 91.4%, and for miRNA-144, AUC was 0.86 (0.69-0.93), with a sensitivity of 88.7% and specificity of 87.8% at best cutoff values as shown in The strategy of ROC analysis was used to diagnose diabetes in adult patients with moderate and poor glycemic control status. In T2D patients with moderate glycemic control status, the current circulating miRNAs, miR-146a and miR-144; myokines, adropin and irisin; and serum OPN yielded a range of 76.2-91.5% specificity and 71.6–89.4% sensitivity. In T2D patients with poor glycemic control, the studied biomarkers provide a good range of 79.4-94.5% specificity and 76.5–91.4% sensitivity. These best cutoff values additionally recommend the potential use of adropin, irisin, and OPN as diagnostic biomarkers for diabetes with varying glycemic control status.In this study, circulating miRNA levels in serum samples from adult patients with T2D with moderate and poor glycemic control status are significantly associated with the parameters of diabetic controls, HbA1c, FINS, and HOMA-IR, respectively. In the present study, two miRNAs, miR-146a and miR-144, were identified with expression levels that were influenced by disease progression and increased serum levels of inflammatory cytokines, myokines, and osteopontin (OPN). In recent studies, the role of miRNAs was explored and showed to be associated with the pathogenesis of diabetes mellitus in all ages , 62, 63,β, and TNF-α and serum CRP levels were associated with the progression of the T2D disease. IL-6, IL-8, IL-18, IL-1β, TNF-α, and CRP levels were significantly increased in T2D patients with poor glycemic control (>8% HbA1c) compared to patients with moderate glycemic control (>7-8% HbA1c). Respective of gender, the levels of inflammatory cytokines significantly increased in females with type 2 diabetes compared to males of the same group.In the same study population, serum levels of inflammatory cytokines, IL-6, IL-8, IL-18, IL-1α, and CRP levels significantly increased and were detrimentally respective to the risk and the development of type 2 diabetes mellitus. In addition, these inflammatory circulating markers significantly increased in patients with featured insulin resistance and those who were clinically with overt type 2 diabetes mellitus [α are significantly correlated with HbA1c and diabetic nephropathy [Previously, a state of the innate immune system was represented in type 2 diabetes mellitus which significantly leads to an increase in the response of cytokine-mediated acute phase . Thus, mmellitus –22. Alsohropathy –22.α might be associated with increased vascular risk in patients with T2D [α. Also, chronically elevated levels of inflammatory cytokines such as CRP, TNF-a, IL-6, and IL-1b could enhance insulin resistance (IR), disrupt insulin sensitivity, and consequently impair glucose homeostasis resulting in an increase in the risk of T2DM [In patients with hyperglycemic spikes, increased secretion levels of cytokines such as IL-6 and TNF-with T2D –68. In a of T2DM –75.α, IL-1b, IL-18, and CRP significantly contributed to the pathogenesis of T2DM through interfering with the insulin signal and impairing β-cell function [In our study, compared to control nondiabetic subjects, increased inflammatory cytokines were predictors of the onset of T2DM in both men and women, whereas differences in the diabetes risk between sexes depend mainly on the change in the expression levels of cytokines , 77. Likfunction  and actifunction  as well function .Osteopontin (OPN) is a multifunctional protein significantly expressed in different biological cell types including bone cells . It was β, TNF-α, and serum CRP levels, respectively, in T2D patients with poor compared to that of moderate glycemic control status. Previous studies reported an increase in the serum levels of OPN in diabetic patients compared to controls, which significantly contributed and was highly induced by elevated glucose and HbA1c levels [Thus, in this study, serum OPN levels were measured in the serum of T2D patients with both moderate and poor glycemic control status. The results showed that serum levels of OPN significantly increased in patients with poor glycemic control compared to that of moderate glycemic control, respectively. However, serum OPN is highly expressed in the serum of women with T2D compared to men of the same group. In poor glycemic control patients, although females showed higher serum OPN levels than males, the results are statistically nonsignificant. In addition, increased OPN serum levels are significantly correlated with HbA1c, FINS (mUI/mL), and HOMA-IR and cytokines IL-6, IL-8, IL-18, IL-1c levels . The incc levels .β, TNF-α, and serum CRP have been implicated in hypertension associated with diabetes via mediating the vascular effects of both angiotensin II (Ang II) and aldosterone, respectively [Additionally, OPN with other reported inflammatory cytokines particularly IL-6, IL-8, IL-18, IL-1ectively . In manyectively –93.New muscle-secreted cytokines particularly irisin and adropin referred to as myokines are able to regulate glucose, lipid levels, insulin sensitivity, and low-grade inflammation –96. The p = 0.001) reduced in T2D patients with poor compared to that of moderate glycemic control status. Also, a higher significant change (p = 0.001) in irisin and adropin was reported in women compared to men with diabetes.In our diabetic patients, the levels of both adropin and irisin are significantly  values, adiposity, inflammation, cytokines, and insulin resistance could be used as parameters to classify normoglycemic individuals who developed T2D over 10 years [In previous studies, baseline levels of several miRNAs which are significantly correlated with HOMA-IR, 10 years , 137–142In this study, ROC analysis additionally was performed to explore the potential use of miR-146a, miR-144, adropin, irisin, and OPN expression levels as diagnostic biomarkers for both moderate and poor glycemic controlled patients. The AUC values, 0.81, 0.78, 0.82, 0.78, and 0.91, respectively, with a range of specificity (76.2-91.5%) and sensitivity (71.6–89.4%) were identified. In addition, AUC values of the same biomarkers, 0.91, 0.86, 0.87, 0.82, and 0.96, respectively, with a range of specificity (79.4-94.5%) and sensitivity (76.5–91.4%) were identified in T2D patients with poor glycemic control. These best cutoff values additionally recommend the potential use of adropin, irisin, and OPN as diagnostic biomarkers for diabetes with varying glycemic control status.In this study, molecular expression of certain microRNA species, such as miR-146a and miR-144, was identified and significantly associated with the parameters of disease severity, HbA1c, inflammatory cytokines, myokines, and serum osteopontin in T2D patients with moderate and poor glycemic control. The AUC cutoff values of circulating miRNAs, miR-146a and miR-144; myokines, adropin and irisin; and serum OPN were significantly identified by ROC analysis which additionally recommends the potential use of these biomarkers: miR-146a, miR-144, adropin, irisin, and OPN as diagnostic biomarkers with considerable specificity and sensitivity for diabetes in patients with varying glycemic control status.


Elettaria cardamomum (cardamom) is a rich source of phenolic compounds, volatile oils, and fixed oils. Cardamom and its pharmacologically effective substances have shown broad-spectrum activities including antihypertensive, anti-oxidant, lipid-modifying, anti-inflammatory, anti-atherosclerotic, anti-thrombotic, hepatoprotective, hypocholesterolemic, anti-obesity, and antidiabetic effects. This review aims to highlight the therapeutic effects of cardamom on MetS and its components including diabetes, hyperlipidemia, obesity, and high blood pressure as well as the underlying mechanisms in the management of MetS. Finally, it can be stated that cardamom has beneficial effects on the treatment of MetS and its complications.Metabolic syndrome (MetS), as a health-threatening factor, consists of various symptoms including insulin resistance, high blood sugar, hypertension, dyslipidemia, inflammation, and abdominal obesity that raise the risk of diabetes mellitus and cardiovascular disease. Cardiovascular diseases are important causes of mortality among the world population. Recently, there has been a growing interest in using phytomedicine and natural compounds in the prevention and treatment of various diseases. The data was gathered by searching various standard electronic databases  for English articles with no time limitations. All  Metabolic syndrome (MetS), as a metabolic disorder, includes a set of major symptoms such as impaired glucose level, dyslipidemia, central obesity, overweight, insulin resistance, and high blood pressure that lead to cardiovascular diseases (CVD), cancer, diabetes mellitus type 2 (DMT2), short lifespan, and low-quality life. MetS has increased concerns about public health problems throughout the world population . It is m, saffron, rosemary, garlic, and rutin have long been used to treat various disorders , India (25.6%), Kuwait (24.8%), and Australia 22.1%) -10. MetS4%, Indiaisorders -15. Geneisorders . Many ofianum L. , Nigellaa sativa , 19, All sativum , Persea mericana , Solanumelongena , and Ber.1% -10. Elettaria cardamomum also known as cardamon, belongs to the Zingiberaceae family. It is native to the Indonesia and Indian subcontinent, Pakistan, Burma, Bangladesh, tropical and subtropical Asia. It is perennial, herbaceous monocots with 4-5 m in height. Cardamom flowers are the whitish lip located at the tip of the corolla tube , TG, LDL, and HDL in several investigations.Clinical trialsA clinical trial was carried out on obese or overweight pre-diabetic women taking 3 grams of cardamom for 2 months. The obtained results disclosed that mean TC (from 192.6 to 183.7 mg/dl) and LDL-C (from 118.1 to 110.5 mg/dl) significantly reduced. It also showed a protective effect on HDL-C (from 44.1 to 42.7 mg/dl) amount in pre-diabetic subjects . AnotherIn vivo studiesPre-clinical research reported that the administration of cardamom powder suspension in 2% gum acacia  decreased the levels of TC and TG, and increased HDL levels in rats with dexamethasone-induced hepatic steatosis . In an aIn vitro studiesin vitro study indicated that 1,8-cineole, a constituent of cardamom, exhibited anti-oxidant properties in lipoprotein metabolism and reduced lipid accumulation in THP-1 cells  are a well-conserved family of class III deacetylases . In mammThe anti-obesity effects of cardamom and its active constituents have been stated in numerous studies .Clinical trialsAn investigation suggested that the administration of cardamom  caused an increase in SIRT1 (from 1.2 to 1.3 ng/ml) in the non-alcoholic fatty liver patient . AnotherIn vivo studiesIt was also found that administration of cardamom powder  prevented obesity in high-fat diet-induced obese rats .Effect of E. cardamomum on hypertensionHypertension is a metabolic risk factor for CVD. It raises the possibility of numerous CVDs, such as peripheral vascular disease, heart failure, coronary artery disease, and stroke . Several lines of evidence have revealed that cardamom and its active constituents show hypotensive and cardiovascular protective properties.Clinical trialsA study showed that treatment with cardamom  caused a significant decrease in systolic blood pressure (SBP) (from 154.2 to 134.8 mmHg) and diastolic blood pressure (DBP) (from 91.8 to 79.6 mmHg) in individuals with primary hypertension of stage 1. Also, cardamom ameliorated ﬁbrinolysis and improved anti-oxidant status after 3 months . In contIn vivo studiesin vivo study showed that cardamom crude extract (3-100 mg/kg) reduced blood pressure by a combination of cholinomimetic and calcium channel blocking mechanisms in rats  was able to attenuate mean arterial pressure in hypertensive rats. The authors suggested that α-terpineol can decrease arterial pressure, Probably by decreasing vascular resistance and repairing enzymatic anti-oxidants in these animals . These fIn vitro studiesin vitro study on rabbit aortic endothelial cell line revealed that α-terpineol  increased nitric oxide (NO) levels via NO–cGMP pathway activation (The vasodilatory effect of cardamom (3-10 mg/ml) has been shown against contraction induced by phenylephrine (1 µM) and potassium (80 mM) in the isolated rat aorta . An in vtivation . As a result, different mechanisms such as calcium channel blocking, induction of NOS and endothelial NO, cholinergic effect, and anti-oxidant property are included in the cardioprotective and hypotensive effects of cardamom and its active ingredients . Based oEffect of E. cardamomum on hyperglycemiaHyperglycemia is one of the risk factors of metabolic syndrome. Hyperglycemia can induce vascular inflammation , microvaE. cardamomum and its active constituents can control insulin secretion, insulin resistance through increasing the amount of SIRT1, PPAR-γ coactivator-1 alpha (PGC-1α), and attenuating the function of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) as well as controlling glucose metabolism by inhibiting α-glucosidase and α-amylase.Cardamom can ameliorate high blood glucose, insulin resistance, and glucose metabolic disorders. Relevant studies regarding the effect of cardamom on diabetes, insulin resistance, and glucose metabolism will be discussed below.Clinical trialsA clinical trial demonstrated that administration of cardamom  to a non-alcoholic fatty liver patient caused an increase in SIRT1 (from 1.2 to 1.3 ng/ml) . SIRT1 iIn vivo studiesIt has been shown that oral administration of cardamom suspension ) reduced hepatomegaly and hyperglycemia in Albino rats . NanolipE. cardamomum extract  for 8 weeks in diabetic rats reduced insulin resistance in animals’ brains by increasing glutamate receptor expression (α-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) glutamate receptor 1 (GluR1) subunit and N-methyl-D-aspartate receptor (NMDA) receptor subunits NR1, NR2A, NR2B)  .In vitro studiesin vitro study on aqueous and methanolic cardamom extracts showed that these extracts have α-glucosidase and α-amylase inhibitory effects and can be used in managing diabetes  in the treatment of T2DM and could be tested in clinical trials . The hyddiabetes .It can be suggested that cardamom and its active ingredients disclose their therapeutic or preventive properties against diabetes via several mechanisms including decreasing glucose level, insulin resistance, increasing insulin level, glucose uptake, anti-oxidant eﬀects, and the number of β-cells in the pancreas  Figure . HoweverE. cardamomum and its active constituents in ameliorating and attenuating the components of MetS including dyslipidemia , obesity (through increasing the amount of SIRT1 and controlling some genes involved in obesity), hypertension , and high blood glucose  that have been reported in clinical trials, in vivo, and in vitro documents. Although there are growing numbers of animal studies demonstrating positive effects of cardamom in MetS, the number of studies linked to cardamom protective properties in humans is still limited. This review article also suggests that cardamom may be implicated as a preventative or therapeutic drug against MetS after being examined in different randomized clinical trials.This manuscript reviewed the chief aspects of MetS and protective mechanisms of HH and GK Study conception and design; RY Acquisition of data; RY Wrote the manuscript in consultation with MR and MGR; HH Critical revision.The authors express no conflict of interest.


Only a small fraction of proteins in plants and animals are classified as allergens. The allergenic properties are frequently attributed to certain functional characteristics of the proteins, such as a role in the plant defense against biotic and abiotic stress, to achieve the systematic acquired resistance. In line with this, eight members out of 17 functional pathogenesis-related (PR) protein families have been characterized as allergens. The present review summarizes the molecular features and allergenic significance of allergens of the PR-1 family. Not many allergens have been identified as belonging to this protein family, with most of them having a pollen origin, like mugwort or Bermuda grass. Molecular and structural features of allergenic PR-1 proteins are discussed and attributed to their IgE-reactive properties, clinical manifestation, and cross-reactivity among different foods and inhalants. Several allergens have been identified as members of the pathogenesis-related (PR) protein families, PR-2 (ß-1-3 Glucanases), PR-3 and PR-4 (Chitinase), PR-5 (Thaumatin-like), PR-8 (Chitinase), PR-10 (Bet v 1-like), PR-14 (nsLTPs)  or antigen 5 (Ag5) type proteins . These pβα sandwich folded CAP domain of 150 amino acids  and andβα sano acids , 9, 13. no acids , which ano acids . The sizCucumis melo), belonging to the Cucurbaitaceae family, has been reported in the pollen food allergy syndrome, in particular in ragweed and grass pollen allergic patients  was the first PR-1 protein characterized as an allergen (ACB45874), but never included to the World Health Organization and International Union of Immunological Societies (WHO/IUIS) Allergen Nomenclature database, and therefore in this review named as Cuc m PR-1. The cDNA sequence encodes for a 151 aa polypeptide with a theoretical pI of 9.47 and a predicted molecular mass of 16.97 kDa , Cuc m 2 (profilin), and Cuc m 3. Cuc m 3.0101  is a major source for allergic reactions with a high prevalence of IgE-sensitization in grass pollen allergic patients, ranging from 40% up to 91% in India and Australia, respectively : Cyn d 1 (beta-expansin), Cyn d 7 , Cyn d 12 (profilin), Cyn d 15 and Cyn d 23, Cyn d 22 (enolase), and the PR-1 protein Cyn d 24. Shen et al. showed the abundance of an IgE-reactive 21 kDa protein in Bermuda grass pollen, with a frequency of 29% of specific IgE-binding in Bermuda grass allergic patients (Hordeum vulgare (P35793). Glycoprotein staining indicated a glycosylation of the PR-1 protein. Finally, natural Cyn d 24 was purified (AAU15051.1). Cyn d 24 consists of 153 aa with an additional signal peptide of 97 aa and it displays an SCP-like extracellular protein domain. Remarkably, Cyn d 24 reveals higher aa identities to SCP proteins than to CRISPs, but low aa-identity to group allergen 5. It contains the conserved region GHYTQVVW, which is responsible for plant defense-related activity , wi, wiArtemN-glycosylation reported in the nArt v 2  has been recognized as an important source of allergen, and together with Artemisia argyi (Art ar 2) and Artemisia sieversiana (Art si 2), they are recommended for immunotherapy in Chinese allergic patients  allergens are present in fruit, pollen, leaves, and branches, and can induce systemic, respiratory, gastrointestinal, and cutaneous symptoms. Although classified as entomophilous, like other pollens, recent evidence indicates that in areas with peach tree orchards, pollen grains can be transported by the wind for short distances, facilitated by the presence of pollutants or the disruption of pollen in smaller submicronic particles , a PR-1 protein  , Pru p 2 protein .In a study analyzing 685 children and adolescents between 3 and 19 years and 310 adults from the population, directly or indirectly exposed to peach cultivars, the sensitization to peach tree pollen was estimated to be 20%, being the third most prevalent pollen after olive tree (33%) and grass pollen (26%) , 44. AccPru p 9 is a 138 aa polypeptide with a theoretical pI of 5.11 and a molecular mass of 18 kDa. This allergen is responsible for the induction of respiratory and ocular symptoms in allergic subjects, independently whether they are directly or indirectly exposed. Reported in another study by Blanca et al., from 62 cases sensitized to peach tree pollen, 41% were positive in SPT to Pru p 9 and presented Pru p 9-specific IgE-antibodies. Besides, the nasal provocation tests showed a decrease in the nasal cavity volume greater than 20% with the new aeroallergen described , associaVespa magnifica Vesp ma 5 (Tabanus yao) (Vespula maculifrons) (Vespula vulgaris) (Dolichovespula maculata) (Vespa crabro)  allergens from the venom of hymenoptera, such as the CAP domain forming a potential CAP cavity for ligand binding. Ag5 showed only moderate aa-ids with Pru p 9: 18% with horsefly Ag5 (Tab y 5), 24% with Savannah tsetse fly Ag5 (Glo m 5), and up to 35% with Vesp ma 5. The Ag5 protein family includes several allergens from hornets, wasps, horseflies, and fire ants: The 25 kDa wasp esp ma 5  shares 2nus yao) , Vesp m lifrons) , Ves v 5ulgaris) , Dol m 5aculata) , and Ves crabro) , respectdatabase . AccordiVespoidea species , an invasive weed from the Asteraceae family, has been reported as a respiratory allergen. Recently, a pollen-derived PR-1 protein of 20 kDa, which displays IgE-reactivity with sera from feverfew sensitized patients' from India, has been identified by LC-MS/MS and 2D-LC-MS/MS (Feverfew LC-MS/MS . HoweverHumulus japonicus), an important cause of weed pollinosis in East Asia, is recognized by IgE from 3.4% (1/29) of Japanese hop allergic patients with a positive case history and IgE-sensitization to Japanese hop , Cuc m PR-1 (aa33), and Cyn d 24 (aa25) have only one potential glycosylation site, as predicted using the NetNGlyc – 1.0 server (52), Cyn d 24 and Cuc m PR-1 are conserved between their AA sequence and therefore, could be influencing or be responsible for IgE crossreactivity reactions through crossreactive carbohydrate determinants (The effect of posttranslational modification of PR-1 proteins by carbohydrate determinants needs to be further explored. Pru p 9 contains two potential glycosylation sites at aa0 server . Interesn d 24 aa have onlrminants . SimilarThe clinical relevance of the PR-1 family is partially known. Only few allergens from this widely distributed protein family have been deeply studied, such as Cyn d 24 with 63% of recognition by patients allergic to Bermuda grass pollen and Art v 2 with 63.2% of positive response by patients sensitized to mugwort pollen, or Cuc m 3, a PR-1 food allergen identified in melon allergic patients. In a more complex way, the Ag5 group, encompassing 26 PR-1 allergens, is implicated in hymenoptera allergy, which triggers life-threatening reactions. According to WHO/IUIS Allergen Nomenclature database criteria, all of them could be considered as relevant allergens in their corresponding sources. An important feature of PR-1 proteins is their role as occupational allergens. Of particular interest is the PR-1 protein described in peach tree pollen, Pru p 9, with 43% of IgE binding in patients allergic to this source. In this way, Pru p 9 has been identified as a relevant allergen with critical implications for people working with peach tree cultivars, developing respiratory symptoms .Nevertheless, one of the major difficulties encountered by researchers in studying allergens belonging to the PR-1 protein family lies in the difficulty for identifying cases sensitized, and also for obtaining pollen grains for molecular analysis of their allergenic profile. Probably, this is one reason for the low number of PR-1 allergens identified and registered in databases. However, it is very likely that PR-1 proteins from other sources display allergenic properties. Exploring new PR-1 proteins implicated in allergenicity, and a complete understanding of their structures and IgE-binding epitopes are necessary for improving the diagnoses and treatment in these subjects, reaching precision medicine in allergy.LM-P, AW, and SS conceived and supervised the review topics. LM-P, AW, SS, MB, MS, and NB-L wrote the first draft. All authors contributed to the article and approved the submitted version.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.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Candida albicans, which can cause superficial and life-threatening systemic infections, is the most common opportunistic fungal pathogen in the human microbiome. The two-component system is one of the most important C. albicans signal transduction pathways, regulating the response to oxidative and osmotic stresses, adhesion, morphogenesis, cell wall synthesis, virulence, drug resistance, and the host–pathogen interactions. Notably, some components of this signaling pathway have not been found in the human genome, indicating that the two-component system of C. albicans can be a potential target for new antifungal agents. Here, we summarize the composition, signal transduction, and regulation of the two-component system of C. albicans to emphasize its essential roles in the pathogenesis of C. albicans and the new therapeutic target for antifungal drugs. Candida albicans is the major pathogenic agent for candidiasis and one of the most common conditionally pathogenic polymorphic fungi from the human microbiome. It colonizes multiple ecological niches, including the oral cavity, reproductive mucosa, and the respiratory and gastrointestinal tract of healthy individuals. C. albicans can also cause cutaneous and mucosal infections such as thrush, vaginal infections, and life-threatening invasive infections [Candida collected from around the world for 20 years (1997–2016) in the SENTRY Antifungal Surveillance Program, 46.9% were C. albicans [C. albicans-induced candidaemia in China is 40.1% and up to 69.8% in Norway [C. albicans in ventilator-associated pulmonary candidiasis in ICU patients is as high as 46.36% [C. albicans is even one of the most common coinfection fungi in COVID-19 patients [The infection and mortality rate of candidiasis have significantly increased in recent years due to tumor chemoradiotherapy, the widespread use of antibiotics, and the increase in the number of immunocompromised patients, such as those with HIV infection ,2. Candifections . Among tn Norway . The pro 1997–201 in the SC. albicans colonization of different host niches depends on the capability to sense multiple environmental signals and then regulate its adaptation and switch between colonization and pathogenesis. C. albicans can transform reversibly between yeast, pseudohyphae, and hyphae forms, adapting to the stresses at different host niches and infected tissues under different conditions, including nutrition, pH value, temperature, oxidation, and immune status. C. albicans possesses a powerful signal transduction network, “the two-component system,” to continuously monitor the external environment and regulate its colonization and pathogenesis [C. albicans regulates morphogenesis, responses to oxidative and osmotic stresses, quorum sensing, virulence regulation, etc. Here we summarize and discuss the structure and function of the two-component system in C. albicans, highlighting its role in pathogenesis and as a therapeutic target for new antifungal agents.The ogenesis . In the Escherichia coli. It is a typical one-step two-component system as an HPK is autophosphorylated on a histidine residue, and the signal is subsequently transferred to an RR on an aspartate residue  and NtrC (an RR protein). NtrB catalyzes the transfer of a phosphate group to the aspartic acid of NtrC under nitrogen limitation conditions. The phosphorylated NtrC activates the transcription of nitrogen metabolism genes [Both prokaryotes and eukaryotes, including fungi, myxomycetes, and plants, contain the two-component signaling system , which c residue . This nism genes . On the sm genes ,22. In tsm genes .Figure 1C. albicans contains three HPKs [Cryptococcus neoformans has seven HPKs [Neurospora crassa expresses eleven HPKs [C. albicans and other fungi [In most eukaryotes, the two-component system is a multi-step phosphate transduction system  Figure , usuallyree HPKs , Cryptocven HPKs , and Neuven HPKs . The traer fungi ,36, reguer fungi  : Sln1p, a homolog of Saccharomyces cerevisiae Sln1, was initially identified in C. albicans by Nagahashi et al. [S. cerevisiae SLN1 null strain, indicating that the function of SLN1 is similar to ScSLN1. Nik1p/Cos1p contains 1081 amino acids and is an apparent ortholog of group III histidine kinases [N. crassa), BcBos1p (Botrytis cinerea), AbNik1p (Alternaria Brassicicola), NcNik1p (Parastagonospora nodorum), and ChNik1p (Cochliobolus heterostrophus) also belong to this class [A. brassicicola [P. nodorum NIK1 deletion reduced asexual sporulation in vitro [i et al. . Sln1p ci et al. , with tw kinases ,54–57. Tis class . Nik1p his class ,61. The is class ,63. Althis class ,55. The sicicola . P. nodoin vitro . Chk1p iin vitro .C. albicans (Ypd1pIntermediate transfer protein of ): Ypd1p serves as an intermediate transfer protein to transfer phosphate groups from HPK to RR and YPD1 can complement the S. cerevisiae YPD1 mutation defected functions [YPD1 is the central molecule of the two-component system, and a decrease in YPD1 activity is expected to compromise fungal fitness, virulence, and viability [YPD1 inhibition is fatal to S. cerevisiae and C. neoformans [C. albicans can adapt to the continuous activation of Hog1-MAPK triggered by YPD1 deletion, actively reducing the level of phosphorylated Hog1 [YPD1 seems to be different among fungal species.unctions ,66. Ypd1unctions  and encounctions ,67–69, biability . YPD1 inoformans ,70–72. Hted Hog1 , indicatC. albicans : Ssk1p is a structural homolog of both S. cerevisiae Ssk1p and Schizosaccharomyces pombe Mcs4p [C. albicans [S. cerevisiae. When cells receive thermal or oxidative stimulation, the signal is transmitted along Sln1p-Ypd1p, eventually phosphorylating Skn7p to regulate gene expression [C. albicans, Skn7p plays an essential role in oxidative stress and morphogenesis, but it has less effect upon the maintenance of the cell wall and the osmotic stress response [CRR1/SRR1 is a newly discovered RR in the CUG branch of Candida [C. albicans and plays an important role in virulence, morphogenesis, apoptosis, osmotic and oxidative stress, etc. [be Mcs4p . Ssk1p ialbicans ,73,74. Spression ,68,75–77response ,78–80. C Candida ,81–83. B Candida  reported Candida  showed tss, etc. ,83, indiC. albicans cells [C. albicans [CHK1 mutants, including the truncation of mannan oligosaccharide and β-1,3-glucan (shortened by about 50%) and β-1,6-glucan (increased about four-fold) levels [Candida was lower when cell wall mannan was added, suggesting that the changes of glucan and mannan in CHK1 mutants might lead to enhanced PMN response [SSK1 and CHK1 mutants to the reconstituted human esophageal tissue (RHE) was lower than that of wild-type strains [CHK1 mutants and the down-regulation of Als1p [SSK1 mutants [SLN1 and NIK1 mutants altered the transcription levels of some N- and O-mannosyltransferases, suggesting their role in cell wall assembly and maintenance [The cell wall is the main organelle of fungi, which determines its viability, cell shape, and interactions with the environment, especially in mediating adhesion and host immune response ,85. The ns cells . The RR albicans ,64,87–89) levels ,90. Inte) levels ,92. The response ,94. The  strains ,42, whicof Als1p  of SSK1  mutants ,42,96. Bntenance .C. albicans, including the expression of adhesion molecules (adhesins and extracellular enzymes), immune escape , and the morphological transformation of yeast to pseudohyphae/hyphae [C. albicans [CPH1 and EFG1), genes encoding the mitogen-activated protein (MAP) kinase signaling pathway (MEK1 and CST20), and genes expressed only during hyphal growth: hyphal cell wall protein (HWP1) and candidalysin (ECE1). A basic correlation has been established between hyphal growth defects and virulence [C. albicans with SLN1 or NIK1 deletion is decreased, while the deletion of CHK1 resulted in loss of virulence, in line with the hyphal defect [NIK1 mutant cultured in 30°C liquid media could not be distinguished from that of the wild-type strain, while the hypha formation of NIK1 mutants was defective on a solid agar plate at 37°C [CHK1 and SSK1 mutants had a hypha-forming defect on medium 199 (pH = 7.5), Spider medium, and serum-mediated solid medium. However, they developed hyphae and flocculate extensively in liquid media, possibly due to the false expression of proteins on the cell surface [CHK1 mutants can also form hyphae similar to the wild type strain but down-regulate the expression of virulence factors in liquid media when co-cultured with oral epithelial cells, indicating its critical role in oral candidiasis [CHK1 mutant formed smooth colonies on solid media probably because the cells aggregated in the colonies and could not grow normally to form the same fuzzy colonies as the wild-type strains [Many factors are believed to be related to the virulence of e/hyphae ,89,97,98albicans . Hypha-sl defect . The yeal defect ,102–106. at 37°C ,55,56,87 surface ,89,107.  strains .SSK1 mutants were defective in hypha development on the nitrogen-rich solid media; however, they formed many hyphae and invaded the solid agar on nitrogen-limited solid media. Therefore, SSK1 might not be required to develop hyphae but might play a role in hypha regulation [TPK1-encoded protein kinase A (PKA) plays a critical role in regulating morphogenesis of C. albicans [TPK1 mutants on solid media was similar to that of SKN7 mutants [SKN7 may be related to TPK1 [SKN7 was also closely correlated with hyphal-specific genes such as CPH1, EED1, EFG1 and UME6 [SKN7 in wild type strains were formed wrinkled colonies and contained filamentous cells. However, the overexpression of SKN7 in CPH1, EED1 or EFG1 mutants did not appear wrinkled colonies and only yeast cells were found, while overexpression of SKN7 in UME6 mutants resulted in slightly wrinkled colonies with yeast cells and pseudohyphae [CPH1, EED1, EFG1 and UME6 are essential for SKN7 function in morphogenesis regulation. SKN7 was closely correlated with hyphal-specific genes such as CPH1, EED1, EFG1, and UME6 [C. albicans are exposed to hypha-inducing solid media, SKN7 is required to limit the accumulation of ROS [C. albicans in liquid media [in vivo [NIK1 mutants is significantly lower than that of the wild-type strain; however, they still form extensive pseudohyphae in tissues [SKN7 mutants is defective, but the virulence does not decrease [PBS2 and HOG1 mutants attenuated virulence in a mouse model of disease [PBS2 and HOG1 positively regulate the virulence of C. albicans. Currently, the regulation of SSK2 on virulence has not been reported.The gulation .The TPK1albicans ,108. The mutants ,80,108, and UME6 . When C.id media . However[in vivo . The vir tissues . Hypha fdecrease ,96. In t disease , indicatC. albicans seem to be tissue-specific [CHK1 and SSK1 mutants are both nontoxic in the disseminated murine model of candidiasis; however, CHK1 mutants are toxic in the rat model of vaginitis [C. albicans gene expression at these two sites. Increased production of neutrophil-dependent lactic acid induces cell wall remodeling, masking critical pathogen-associated molecular patterns (PAMPs), such as glucans, blocking immune recognition, and allowing C. albicans to colonize and invade the host [C. albicans [SSK1 and CHK1 mutants are more susceptible to growth inhibition and killing efficacy of PMNs [SSK1 mutants to human neutrophil defensin-1 (HNP-1) was higher than that of wild-type strains [The virulence and immune system evasion of specific ,112. CHKaginitis ,87,113. the host . PMNs arthe host . Althoug of PMNs ,116. The strains .YPD1 increases C. albicans virulence and its ability to kill macrophages, which might be related to the phenotype of increased hyphae [SKN7 mutants were significantly less susceptible to the killing by PMNs than the SSK1 mutants, and their virulence in the disseminated murine candidiasis model was only mild or not weakened [CHK1 and SLN1, and CHK1 and NIK1 double mutants were survivable, while the SLN1 and NIK1 double mutants could not be constructed, indicating that the pairing loss of these kinases was a fatal event [C. albicans lack of Chk1p was nontoxic in the disseminated murine candidiasis model; however, if CHK1 mutation was accompanied by SLN1 or NIK1 deletion, both hyphal development and virulence of the mutant were enhanced [SSK1 and HOG1 can negatively regulate the expression of CHK1 [SSK1 mutant also down-regulated the expression of the following hypha regulation and virulence factors: HYR1, HWP1, ECE1, MIG1, GCN4, RFG1 (ROX1), RBF1, RIM101, HAC1, HAP5, TUP1, NRG1, EFG1, and CPH1 [It is noteworthy that inhibiting the expression of d hyphae . The SKNweakened . By knocal event . Moreoveenhanced . Deletio of CHK1 ,96, sugg of CHK1   and Ssk1p (Asp). The phosphorylated Ssk1p cannot activate Ssk2p, leading to the shutdown of the Hog1-MAPK cascade. Another situation is that increased extracellular osmolarity deactivates Sln1p without the above-mentioned phosphate transport. A non-phosphorylated form of Ssk1p initiates the downstream signal system and continuously activates Ssk2p/Ssk22p (MAPKKK), Pbs2p (MAPKK), and Hog1p (MAPK). Finally, the phosphorylated Hog1p is transferred to the nucleus, activating transcription factors to induce the GPD1 expression, which increases the intracellular glycerol content to adapt to hyperosmotic stress  are necessary for C. albicans to resist oxidative stresses [SSK1 and SKN7 are more sensitive to a series of oxidants, including H2O2 and t-BOOH in vitro [SKN7 and CRR1 [SKN7 activation by oxidative stresses requires functional mitochondria in S. cerevisiae [YPD1 mutant of C. albicans was highly resistance to H2O2 and t-BOOH, but it was very sensitive to sodium arsenite [Two-component system proteins play an essential role in the oxidative stress response of albicans . Of the e strain . Three Rstresses ,53,80,82in vitro ,53,74,80and CRR1 ,80,82. Srevisiae . The YPDarsenite ,67.FigurC. albicans is similar to the quorum sensing system of some bacterial species [C. albicans exist in the form of yeast when the cell density is more than 106 cells/ml, while hyphae are formed when the cell density is less than 106 cells/ml [C. albicans, which might inhibit biofilm formation by regulating hyphal morphogenesis [C. albicans as SLN1, NIK1, and SSK1 mutants respond to farnesol similar to the wild-type strains, while CHK1 mutants can still form biofilms when farnesol is added [The relationship between cell density and hypha formation of  species . Under tcells/ml . Farnesoogenesis . Chk1p mis added ; howeveris added , as Chk1Candida on the surface of mucous membranes, dentures, central venous catheters and other medical devices can serve as physical barriers to drug or molecular penetration, making Candida inherently resistant to traditional antifungal drugs and host immune responses [SSK1 and HOG1 mutations can restore the susceptibility of clinical strains of the emerging Candida species Candida auris to amphotericin B and caspofungin. The SSK1 and CHK1 mutants of C. albicans were highly sensitive to fluconazole and voriconazole, and the sensitivity of SSK1 mutants to fluconazole was 30 times higher than the CHK1 mutants, while the sensitivity of SLN1 and SKN7 mutants were slightly higher or equal to the wild-type strains [C. albicans and caused cell wall defects by inhibiting the hypha formation and growth. They also significantly enhanced antifungal effects when combined with fluconazole [S. cerevisiae could activate Hog1p in the absence of external stimuli, similar to the effect of bactericide treatment [YPD1 is not fatal to C. albicans and even enhances its virulence [YPD1 as a broad-spectrum antifungal target.Currently, classic antifungal drugs mainly include echinocandins, polyenes, and azoles . Due to esponses . Therefoesponses ,143–146.esponses  reported strains . Both riconazole . The delreatment , suggestirulence . These fC. albicans, and significant progress has been made in the past two decades; however, there are still many aspects to be further explored. For example, how are signals distinguished and transmitted to the downstream different RRs after the intermediate transfer protein received stimulation from different HPKs? What other functions of the Crr1p/Srr1p have not been identified? If Crr1p/Srr1p is present in mitochondria, can the HPK bypass Ypd1 in the nucleus and cytoplasm and directly transmit the signal to the RR? The different regulatory functions of the same two-component system under different conditions and the interaction of the system with other transcription factors or signaling proteins are also needs to be investigated. The components of the signal pathway have not been found in the human genome. The development of new drugs against the two-component system can effectively target fungi without damaging the host cells, to greatly reduce the toxic or side effects of antifungal drugs. The two-component system of C. albicans therefore provides the ideal therapeutic target of for new antifungal drugs against C. albicans.The two-component system plays a vital role in the activities and pathogenicity of


Frailty assessment is recommended before elective transcatheter aortic valve implantation (TAVI) to determine post-interventional prognosis. Several studies have investigated frailty in TAVI-patients using numerous assessments; however, it remains unclear which is the most appropriate tool for clinical practice. Therefore, we evaluate which frailty assessment is mainly used and meaningful for ≤30-day and ≥1-year prognosis in TAVI patients. Randomized controlled or observational studies (prospective/retrospective) investigating all-cause mortality in older (≥70 years) TAVI patients were identified . In total, 79 studies investigating frailty with 49 different assessments were included. As single markers of frailty, mostly gait speed (23 studies) and serum albumin (16 studies) were used. Higher risk of 1-year mortality was predicted by slower gait speed (highest Hazard Ratios (HR): 14.71; 95% confidence interval (CI) 6.50–33.30) and lower serum albumin level . Composite indices  were associated with 30-day (highest Odds Ratio (OR): 15.30; 95% CI 2.71–86.10) and 1-year mortality . In conclusion, single markers of frailty, in particular gait speed, were widely used to predict 1-year mortality. Composite indices were appropriate, as well as a comprehensive assessment of frailty. The proportion of older people (>65 years) is predicted to constantly increase in Europe and reach around 30% of the total population by 2060 . These sFrailty is a complex clinical condition , where pOlder TAVI patients are characterized by decreased physiological functionality , multiple comorbidities and malnutrition, with the prevalence of frailty found to be up to 63% in this population ,6,10. PrTherefore, the aim of our review was to evaluate which frailty assessment is the most commonly used and meaningful for the prediction of short-term, intermediate-term and long-term all-cause mortality in older TAVI patients. The electronic database PubMed (May 2020) was systematically searched. Study inclusion criteria were the following: randomized controlled trials (RCTs) or observational studies  without time restrictions that quantitatively assessed frailty pre-interventionally in older (≥70 years) patients who underwent elective TAVI. The outcome of interest was the association of frailty with all-cause mortality. Identification, screening, eligibility and inclusion processes were performed according to PRISMA guidelines. The included articles were reviewed by two independent researchers. The overall risk of bias in each study was evaluated using the Quality in Prognosis Studies (QUIPS)  and the n = 79 studies were included for full-text review , blood parameters , impairments in activities of daily living (ADL), nutritional status/risk , cognitive impairments  and body composition (Psoas muscle area index), as shown in In total, t review  and n = In n = 4 prospective [n = 6 retrospective [n = 7 prospective [n = 9 retrospective [Gait speed  was the spective ,20,32,33spective ,31,34,35spective  showed ispective  and Alfrspective ,28,37,39spective ,30,34,38spective ,28,29,30n = 6 retrospective studies [n = 4 prospective [n = 10 retrospective [Serum albumin level was the second most commonly used single marker of frailty and was associated with short-term mortality in  studies ,34,35,40spective ,39,43,46spective ,42,44,45spective ,34. n = 3 prospective [n = 4 retrospective [n = 9 prospective [n = 6 retrospective [Activities of Daily Living (ADL) were evaluated in spective ,51,52   were ass studies ,51,54,55 studies ,51, inte studies ,54 or lo studies ,22,55 mo studies ,51 of ren = 3 prospective [n = 5 prospective [n = 1 retrospective [n = 2 studies [The Mini-Mental State Examination questionnaire  was usedspective ,33,51 stspective  showed aspective ,22,51,55spective   was investigated in only  studies ,57 with  studies  found thspective ,57,58,60spective ,45,59 (2n = 7 prospective studies [The Timed Up-and-Go test (TUG) was used in  studies ,51,54,55 studies  to 344 [ studies  patients studies  found li studies  found thn = 9 studies [n = 2 prospective [n = 1 retrospective [n = 4 prospective [n = 4 retrospective [The Clinical Frailty Scale (CFS)  was used studies ,63,64,65spective ,32   was not spective ,25,26  was used in spective ,37,74,75spective   and malnutrition . Patients with a slower walk speed (≥6 s) measured by the 5-mWT showed a higher relative risk of 1-year mortality. Pre-interventional lower levels (≤3.5 g/dL) of serum albumin were associated with a worse long-term prognosis as well. Further, higher scores of composite indices  were associated with higher mortality risk over the short and long term after intervention. n = 23) investigated gait speed with the 5-mWT and found an association with higher 1-year mortality in older TAVI patients. Slower gait speed is largely used to evaluate impaired health conditions, especially among the elderly [A great number of studies  . InitialIn this review, we provide a detailed overview of the frailty assessments most commonly utilized in clinical practice, divided by category and their ability to predict mortality risk. However, due to the remarkable heterogeneity of the included studies, meta-analyses of the study results were not indicated. Therefore, this review does not provide pooled effect sizes for the frailty assessments investigated, potentially limiting information for the clinician. Heterogeneity is notable especially in regard to the methodology of frailty measurements, study designs and statistical analyses. For example, seven different cut-off values were identified for gait speed. This variety of cut-off values might have influenced the estimation of patients considered as frail across the studies. For composite indices, different measurement approaches  were evident across studies. Additionally, the lack of a general agreement on the definition of frailty  and the First of all, there are two fundamentally different approaches to conceptualizing frailty. On the one hand, Fried et al. (2001) describe frailty based on the phenotype, focusing on physical components  . One theAlthough frailty is a multidimensional syndrome among older adults characterized by a marked vulnerability and diminished capability to recover from stressors, a considerable number of single markers of frailty were used in the included studies. From the statistical point of view, in multivariable analyses of mortality, single parameters often show equivalent or stronger predictive effects than composite measures ,88,89. TFor patients scheduled for TAVI and classified as frail, the effect of prehabilitation programs on morbidity and mortality is currently being investigated . Taking Frailty was most commonly assessed using single markers of frailty, especially based on measurements of gait speed (5-mWT) and pre-interventional levels of serum albumin. Slow gait speed (≥6 s) and a level of serum albumin ≤ 3.5 g/dL were predictive of a higher risk of 1-year mortality in older TAVI patients. Composite indices (5 items), considered as a comprehensive assessment of frailty, were associated with short- and long-term mortality. However, considerable heterogeneity was observed among the studies, and the methodology of each frailty assessment should be carefully considered.


H3K27M-mutant midline gliomas have variable clinical presentations, prognoses, and molecular backgrounds. In this study, we integrated data from published studies to investigate the differences between these two groups.Pediatric and adult H3K27M-mutant midline gliomas. For time-to-event analyses, Kaplan-Meier analysis and Cox regression models were carried out; corresponding hazard ratios (HR) and 95% confidence intervals (CI) were computed to analyze the impact of age and clinical covariates on progression-free survival (PFS) and overall survival (OS).PubMed and Web of Science were searched for potential data. Studies were included if they had available individual participant data on patients age of H3K27M mutation for analyses. In adults, there was a male predilection whereas females were slightly more common than males in the pediatric group. Spinal cord tumors were more frequent in adults. The prevalence of H3.1 K27M mutation was significantly higher in the pediatric cohort. Compared to adult patients, pediatric H3K27M-mutant midline gliomas exhibited more aggressive features including higher rates of pathologic features of high-grade tumors and Ki67 proliferation index, and had a shorter PFS and OS. Genetically, ACVR1 mutations were more common whereas MGMT methylation, FGFR1, and NF1 mutations were less prevalent in the pediatric cohort.We included 43 studies comprising 272 adults and 657 pediatric midline gliomas with H3K27M-mutant midline gliomas were demographically, clinically, and molecularly distinct from adult patients, highlighting an opportunity to refine the risk stratification for these neoplasms.Pediatric  H3K27M-mutant glial tumors arise predominantly in midline structures such as the thalamus, brainstem, and spinal cord , HIST2H3C (H3.2), or H3F3A (H3.3) genes with a lysine to methionine amino acid substitution at codon 27 (K27M) and are mutually exclusive with isocitrate dehydrogenase 1/2 (IDH1/2) mutations . The study protocol was strictly adherent to the recommendations of the Preferred Reporting Items for Systematic Review and Meta-Analysis (PRISMA) statement .H3K27M-mutant gliomas and (ii) studies with patient age available. We excluded studies if they were: (i) not relevant to inclusion criteria; (ii) case reports; (iii) reviews, theses, or books; (iv) conference or proceeding papers; or (v) studies with duplicated populations.Results from the two electronic databases were imported into EndNote  and duplicates were subsequently removed. Next, two reviewers (HGV and TNMN) independently screened the title and abstract of the articles using the following inclusion criteria: (i) studies providing individual participant data (IPD) of H3 genotypes, detection methods, demographic information, tumor size, tumor location, histology grades, Ki67 index, treatments, PFS time, PFS status, OS time, OS status, and genetic alterations of H3K27M-mutant gliomas. Subsequently, we removed cases with missing data of patient age/tumor location or cases from non-midline locations.Two reviewers (HGV and TNMN) independently reviewed the full text of potential studies and extracted data into a standardized worksheet. We also carefully reviewed the reference list of the included studies to find additional papers. The following IPD were extracted from the articles: authors, institution, country, year of publication, study period, patient identification number, H3K27M-mutant gliomas. The deviance residuals and the dfbeta values were used to examine influential observations. Hazard ratios (HR) are presented as mean and 95% confidence interval (CI). A two-sided p-value of < 0.05 was considered statistically significant. The statistical analyses were performed using the R software, version 3.6.1 .Patients were divided into pediatric (patients of 18 or less than 18 years of age) and adult groups (patients of more than 18 years of age). To avoid duplicated data between studies from the same institutions, we selected the study with the largest population. Categorical and continuous variables of pediatric and adult cohorts were compared utilizing Chi-square, Fisher’s exact test, and Wilcoxon rank-sum test, if applicable. Kaplan-Meier analysis and Cox proportional hazards model were conducted to assess the impact of various clinical parameters on survival of H3K27M-mutant midline gliomas for integrated analyses . There was a male predilection in adult H3K27M-mutant gliomas whereas females were more common in the pediatric group (p < 0.001). In the pediatric population, H3K27M-mutant tumors were mostly seen in the brainstem whereas a higher proportion of thalamic and spinal lesions were identified in adult patients (p< 0.001). Adult tumors had a lower Ki67 index compared to the pediatric group (p < 0.001).H3K27M-mutant gliomas were associated with higher rates of gross total/subtotal removal and radiotherapy administration (p < 0.001). In comparison to the adult group, pediatric patients had a worse OS   Figure 2H3K27M-mutant gliomas remained significantly shorter   , only a small number of patients were compared (et al. (H3K27M-mutant midline gliomas carry worse outcome despite more aggressive treatment attempts than in adult patients. Our findings showed that tumors in pediatric patients had more aggressive pathologic features including a significantly advanced histological grade and Ki67 proliferation index, which may explain the poor survival of the pediatric cohort. Hoffmann et al. reported that pediatric DIPG with age of less than 3 and more than 10 years were associated with longer-term survival compared to those aged 3-10 (In this study of compared . Schulte (et al.  reportedged 3-10 . In thisH3 K27M mutation in both adult and pediatric high-grade spinal gliomas (H3 K27M spinal cord tumors as compared to the pediatric population. These results were consistent with published population-level data on high-grade gliomas and glioblastomas of the spinal cord that these neoplasms are most commonly seen in adults (H3K27M-mutant diffuse midline gliomas is independent of their anatomical locations (H3K27M midline gliomas in the pediatric population, regardless of anatomical location.While earlier studies reported a high frequency of  gliomas , we founn adults , 52, as n adults . Severalocations , 53. Howocations , 54. WheH3K27M-mutant midline gliomas were also genetically different. We found that ACVR1 mutations were exclusively seen in pediatric patients, particularly in children less than six years of age. On the other hand, the incidence of FGFR1, NF1 mutations, and MGMT methylation were more significantly prevalent in adult patients. The lower frequency of MGMT hypermethylation in pediatric patients may contribute to the poor response to temozolomide and a worse PFS/OS as compared to adult cohort. FGFR1 mutations are associated with prolonged survival in H3K27M-mutant gliomas (EGFR alterations, TERT promoter mutations, and mutations in the tumor suppressor PTEN are less likely to exist in high-grade gliomas in children (H3K27M-mutated midline gliomas. Hopefully, these observations may help understand the molecular profile of midline gliomas as treatment.Compared to pediatric tumors, adult  gliomas , which mchildren , 57. HowH3K27M-mutant midline gliomas. Because of the rarity of H3K27M-mutant midline gliomas in adult patients, it is challenging to observe the significant differences between pediatric and adult cohorts in institutional studies (H3K27M-mutant midline gliomas. Additionally, there are important clinical factors that we could not incorporate into the multivariate analysis due to missing data, such as the Karnofsky Performance Scale, tumor size, and disease stage. Pediatric patients may have limited self-care ability resulting in lower KPS and advanced disease stages compared with adults. Future prospective studies are needed to confirm the results of this study. Another bias could occur due to differences in treatment protocols across included institutions. Finally, although we strictly screened the included patients using patient identification code, demographic information, and clinical data to avoid duplicating patients, there are still possibilities of overlap among the dataset which may affect the analyses.This study is the first study to demonstrate the differences in demographics, clinical manifestations, prognoses, as well as molecular backgrounds of pediatric versus adult  studies . Our resH3K27M-mutant midline gliomas are demographically, clinically, prognostically distinct from adult tumors. H3K27- mutant midline gliomas in pediatric patients were associated with a significantly shorter PFS and OS. Additionally, the underlying molecular backgrounds of these two cohorts were also different. Our results may improve our biological understanding of H3K27M-mutant midline gliomas.In conclusion, the present study demonstrated that pediatric The original contributions presented in the study are included in the article/HV, conceptualization, data curation, formal analysis, investigation, methodology, project administration, software, validation, writing original, review, and editing. TN, HL, AJ, MH, JB, and RM-K, data curation, formal analysis, investigation, methodology, review, and editing. IFD, data curation, formal analysis, investigation, methodology, project administration, validation, review, editing, and supervision. All authors contributed to the article and approved the submitted version.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.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.