The spectrum of TKI-related adverse events (AEs) is variable. Pleural effusion (PE) is a frequent AE attributable to dasatinib treatment, while it is only rarely associated with nilotinib. The pathogenetic mechanism leading to PE during nilotinib therapy is still unknown and its management has not yet been defined. To the best of our knowledge, only a limited number of similar case reports have already been reported in the literature so far. Here, we describe the case of a 41-year-old CML patient who developed PE during first-line nilotinib, successfully treated with steroids and nilotinib permanent discontinuation. We highlight the differences among our patient and the others, proposing therapeutic strategies to solve this rare but still possible AE, of which physicians should be aware. BCR::ABL1-positive chronic myeloid leukemia (CML) is a myeloproliferative neoplasm with an incidence of 1-2 cases per 100.000 adults, which represents approximately 15% of newly diagnosed cases of leukemia in adults. This disease is characterized by a single reciprocal translocation between chromosomes 9 and 22, resulting in the formation of the Philadelphia (Ph) chromosome. BCR::ABL1 fusion gene encodes a p210 protein (BCR::ABL1) with deregulated tyrosine kinase activity. Knowledge of this translocation was the basis for the development of drugs known as small molecule tyrosine-kinase inhibitors (TKIs)  expressing ‘off-target’ toxicity of TKIs such as in the case of pleural effusion (PE) Table 1 As for nilotinib, it has a peculiar cardiovascular (CV) toxicity reported in 20% of patients, contraindicating its use in case of previous CV diseases, while other common side effects may include hypercholesterolemia and hyperglycemia . In the BCR::ABL1 p210 fusion transcript and the JAK2V617F mutation. The latter was not detected; on the contrary, a diagnosis of CML was made by means of qualitative PCR, which demonstrated the presence of a typical e14a2 (b3a2) BCR::ABL1 p210 configuration. Bone marrow morphological analysis showed a CP-CML pattern, and cytogenetic studies revealed a 46,XY karyotype with the t[20/20], and no additional chromosomal abnormality. After previous debulking therapy with hydroxyurea, considering his young age, absence of any significant comorbidities and CML risk scores, first-line therapy with nilotinib 600 mg BID was started. After 3 months, the patient achieved an optimal response as defined in 2020 ELN recommendations . However, at 6 months the patient complained about weight gain of 7 Kg and dyspnea on exertion, with no significant peripheral edema. An abdomen ultrasound showed neither hepato-splenomegaly nor ascites, while extensive bilateral PE with disventilative compression phenomena was collaterally detected. Chest X-ray confirmed this finding; even though there was no evidence of pericardial effusion in echocardiography, nilotinib was prudently discontinued. In addition, to exclude a concomitant lung infection, a CT scan was performed showing Grade 2 PE according to CTCAE v.4.0, and parenchymal thickening associated with shaded areas of peribronchiolar groundglass and signs of interstitial edema. Broncholwash excluded Aspergillus, respiratory viruses including SARS-CoV2, CMV-DNA, mycobacteria, Legionella pneumophila, Mycoplasma pneumoniae, Chlamydia pneumoniae, and Pneumocystis jirovecii infections. Interestingly, flow cytometry performed on broncholwash documented lymphoid cellularity for 11% distributed as follows: 66% T CD3+, CD4/CD8 = 1.6; 32% NK CD3-/CD16+; 2% B CD19+. No reactivity was instead detected for CD117 and/or CD34. Three days later, once an infectious etiology has been ruled out, steroid treatment with prednisone 25 mg QD was started. After 10 days a new chest X-ray was performed, documenting complete resolution of lung involvement together with an initial improvement in both dyspnea and body weight. Due to the severity of nilotinib toxicity and the optimal response already achieved, the drug was definitively discontinued and imatinib was started at the dosage of 400 mg QD. Imatinib therapy was well tolerated with no significant AE, in parallel with a progressive reduction in BCR::ABL1 transcript level up to 0.7695% IS. However, after 6 months from imatinib start molecular evaluation showed a slight increase in BCR::ABL1 transcript level till to 0.7924% IS, thus defining a warning response according to 2020 ELN recommendations . Accordingly, considering the young age of the patient and the absence of any CV risk factors (score CHART 1%), third-line therapy with ponatinib at the dosage of 30 mg QD was started from September 2021. This new treatment did not lead to any significant toxicity and allowed the patient to achieve a progressive reduction in BCR::ABL1 transcript level, till to the obtainment of a major molecular response (MMR).Here, we describe the case of a 41-year-old man with a history of esophageal achalasia affected by chronic phase (CP)-CML, high risk according to Sokal and low risk according to ELTS score. In July 2020 he was admitted to our hospital because of pain in left hypochondrium. Blood exams showed extreme leukocytosis [289.850/mmc with 5% of peripheral blood (PB) blasts] and thrombocytosis (903.000/mmc), associated with mild anemia and moderate splenomegaly (bipolar diameter of 17.5 cm at ultrasound examination). Given the diagnostic suspicion of a myeloid neoplasm, we searched for the ndations , with a ndations Figure 1The most common non-hematological AEs of nilotinib are already well-known, including peripheral arterial occlusive diseases, followed by QTc interval prolongation, pancreatic enzymes, bilirubin and glucose blood levels elevation, gastrointestinal symptoms, pruritus, rash, headache, fatigue, arthralgia, nasopharyngitis, fever and night sweats; on the contrary, dasatinib has a peculiar pulmonary toxicity with a high incidence of PE estimated between 14% and 30% .These AEs may be due to “off-target” effects; in particular, dasatinib-induced PE may be secondary to potent PDGFR-β inhibition in association with other possible mechanisms such as SRC inhibition . Indeed,One possible explanation is that dasatinib-related PE may be secondary to the cytotoxic T and NK cells expansion or to the action of other kinases . Indeed,vs. imatinib for patients with newly diagnosed CP-CML, PE occurred in 5.2% of bosutinib-treated subjects, with the most promising risk factors for this AE, in addition to bosutinib treatment, represented by advanced age, smoking habit, and history of pulmonary events , individual characteristics of CML subjects, including comorbidities, lifestyle preferences, and TKI compliance, along with distinct ‘off-target’ TKI toxicities (which can lead to drug-related long-term morbidities) and molecular  therapy , 32–34.All this considered, returning to our patient, it was decided not to restart nilotinib, even at half the standard dose, but to introduce imatinib for multiple reasons: firstly, unlike dasatinib and bosutinib, for which it is already known that they can be resumed at the same dose once the first episode of PE has resolved , no specBCR::ABL1 transcript level, in order to avoid new drug suspensions due to recurrences of PE, it was decided not to restart nilotinib, not even at a lower dosage, but to change the TKI by starting imatinib in the light of its safer toxicity profile. Unfortunately, due to an inadequate response to imatinib, once any known ABL1 mutation was ruled out, given the patient’s young age and the absence of any CV risk factors, third-line therapy with ponatinib was started at the dosage of 30 mg QD, allowing the patient to quickly achieve a MMR.In this case report, nilotinib was found to be the only possible cause of PE, after excluding other etiologies. However, unlike previous experiences, due to the severity of this rare AE and also considering the optimal response that the patient had already obtained, with the aim of preventing future PE recurrences, nilotinib was permanently discontinued with no new episodes during subsequent TKIs. In addition, some differences from other cases should be noted: firstly, this AE occurred during first-line therapy without concomitant medications. Overall, considering these experiences, it is not possible to hypothesize clear risk factors for nilotinib-induced PE with the sole exception of male sex. Indeed, differently from gender, older age does not seem to be prognostically relevant. The same was true for comorbidities: the first patient reported had coronary artery disease and hypertension, among others. The remaining patients, including the one described in this case report, had no significant comorbidities. Duration of treatment also does not appear to have an impact on PE risk as patients developed this AE within 2-3 months to 5 years of starting nilotinib. An unmet clinical need may be the best management of this AE: apart from supportive care, i.e., steroids and diuretics, the real indication for switching from nilotinib to another TKI after a single episode of PE is still unclear. In our case, due to the severity of the clinical presentation of this rare AE and the progressive reduction in The original contributions presented in the study are included in the article/supplementary material. Further inquiries can be directed to the corresponding author.Written informed consent was obtained from the individual(s) for the publication of any potentially identifiable images or data included in this article.All authors listed have made a substantial, direct, and intellectual contribution to the work and approved it for publication.This study was partially funded by Italian Ministry of Health - Current research IRCCS.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.


However, pure-tone audiometry (PTA) is an investigation that can be used for the screening of these patients in order to reduce the costs. The aim of this systematic review and meta-analysis was to evaluate the diagnostic accuracy of different PTA protocols for VS in patients with ASHL, when compared with MRI; (2) Methods: Medline, Embase, and Cochrane databases were used to find relevant studies. All prospective and retrospective observational studies that evaluated the accuracy of PTA protocols for the screening of VS were assessed, according to the international guidelines; (3) Results: We analyzed seven studies (4369 patients) of poor-to-moderate quality. Their pooled sensitivity was good (0.73–0.93), but their specificity was low (0.31–0.60). All protocols were located in the right lower quadrant on the likelihood scattergram, and the post-test probabilities for positive and negative diagnosis of these protocols were extremely low; (4) Conclusions: PTA protocols cannot be used for a proper screening or diagnosis of vestibular schwannoma despite their good sensibility, and MRI remains the gold standard for this purpose. The Schwann cells of the vestibular  nerve give rise to the benign tumor known as vestibular schwannoma (VS)/acoustic neuroma (AN). Despite their benign character, these tumors have the ability to grow and can cause severe ontological symptoms, such as unilateral sensorineural hearing loss/asymmetrical hearing loss (AHL), vertigo, and tinnitus, due to impairment of the vestibulocochlear nerve function ,2. GraduA VS typically grows between 0.99 and 1.11 mm every year, but certain characteristics of the tumor, including cystic and hemorrhagic appearances, as well as erythropoietin treatment, have been proven to indicate an accelerated growth . Regarding the epidemiological profile of VS, it was demonstrated that the radiologically confirmed vestibular schwannoma rates increased in recent years in the United States of America  . At the Although a consistent association between long-term mobile phone use and the risk of developing VS has not been documented, there is heterogeneity within investigations, and higher risks have been noted in several studies for use of more than 10 years ,12,13. EMagnetic resonance imaging (MRI) is the gold standard investigation for all patients who present with asymmetrical hearing loss . The useHowever, screening methods have been devised to save and maximize resources because the number of MRI exams required for this group of patients is very high, and the number of schwannomas discovered is relatively low. An objective approach is represented by audiometric protocols based on quantifying the pure-tone audiometric (PTA) threshold by the difference in “decibel” and “frequency regions” between two ears . CurrentThe aim of this systematic review and meta-analysis was to evaluate the diagnostic accuracy of different PTA screening protocols for vestibular schwannoma in patients with asymmetrical hearing loss and to compare it with the gold standard represented by MRI. We performed a systematical search of published studies that evaluated PTA protocols for the screening of VS, in comparison with MRI examination in MEDLINE, EMBASE and Cochrane Library using synonyms of ‘magnetic resonance imaging‘, ‘asymmetrical hearing loss‘, ‘vestibular schwannoma‘, and Boolean operators AND/OR in accordance with Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines . The full-text papers were independently reviewed by two investigators (M.D.C and D.N.) to establish their eligibility for the review. Any differences between the two were remedied by a third reviewer (M.L.C.) if a consensus could not be reached. The inclusion criteria were represented by observational studies, both prospective and retrospective, with a diagnostic study design that compared at least one PTA screening protocol to MRI findings in patients with AHL and comprised sufficient data for a 2 × 2 contingency table creation. We excluded opinion papers, animal studies, and case reports from the search. Two investigators (B.M.C and M.L.C) retrieved data from the eligible studies separately using a standard process. Data concerning the first author, publication year, study design, characteristics of the population examined, number of cases and controls, cut-offs used, and the information needed to create a 2 × 2 table were obtained. Two independent reviewers (B.M.C and M.L.C) assessed the methodological quality of the included studies using the QUADAS-2 technique  . Any dis2 statistic was used to quantify the degree of heterogeneity. The statistical analyses were performed using STATA SE . We summarized data from each study in 2 × 2 tables of true-positive, false-positive, true-negative and false-negative values, and we calculated sensitivity, specificity, and positive and negative likelihood ratios, as well as diagnostic odds ratio. For hierarchical modeling, a hierarchical summary receiver operating characteristic (HSROC) model will be utilized to generate equal summary estimates for sensitivity and specificity, taking into account variability both between and within studies (heterogeneity). In order to show variation and explore heterogeneity for sensitivity and specificity, we drew Forrest plots, likelihood ratios scattergrams, bivariate boxplots, and Fagan nomograms. IOur search yielded 400 unique records, out of which only 7 were included in the meta-analysis after abstract and full text screening . We did The characteristics of the included studies are presented in Overall, the quality of included studies was low-to-moderate . Two stuThe pooled estimates and confidence intervals of sensitivity, specificity, positive and negative likelihood ratios, and diagnostic odds ratio, corresponding to the evaluated PTA protocols are presented in The highest pooled sensitivity was achieved by the following protocols: Mangham 0.93 (95% CI: 0.76–0.98) ,23,24,25The highest pooled negative likelihood ratio corresponded to the Sheppard protocol 0.45 (95% CI: 0.24–0.85) ,21,23,252: 87.4%) and specificity (I2: 98.3%) of this protocol  and specificity (I2: 98.3%) of this protocol  and specificity (I2: 97.2%) of this protocol  and specificity (I2: 96.9%) of this protocol  and specificity (I2: 98.9%) of this protocol  and specificity (I2: 94%) of this protocol  and specificity (I2: 97.2%) of this protocol  and specificity (I2: 98.8%) of this protocol  and specificity (I2: 99.2%) of this protocol  and specificity (I2: 94.4%) of this protocol  and specificity (I2: 93.4%) of this protocol , Amclass (0.93), and Nashville (0.91) protocols. On the other hand, the specificity of the evaluated protocols was heterogeneous and low, ranging from 0.31 (Nashville) to 0.60 (AAO). Our study results revealed good values for the HSROC curve, ranging from 0.66 (Mangham) to 0.92 (Amclass). Nonetheless, all protocols were located in the right lower quadrant on the likelihood scattergram, which indicated that none of them could be used for exclusion or confirmation of the disease. Moreover, the post-test probabilities for positive and negative diagnosis of these protocols were extremely low. These arguments support the hypothesis that the evaluated pure-tone audiometry protocols cannot be used for a proper screening or diagnosis of vestibular schwannoma despite of their good sensitivity. Thus, MRI investigation remains the gold standard for the evaluation of patients with unilateral hearing loss, even though its costs are high ,36,37. Even though PTA protocols could be used in low-resource medical settings due to their high sensibility, simplicity, objectivity, easiness to apply, and low costs, clinicians must take into consideration their low specificity, which may give a high number of false positives when evaluating patients with unilateral hearing loss . MoreoveIn recent years, data collected from PTA along with the patient’s clinical characteristics were incorporated into algorithms for the prediction of the need for active treatment with approximately 90% accuracy . This paOur results are comparable to those reported in a 2017 systematic review and meta-analysis that evaluated the diagnostic accuracy of different non-imaging screening protocols that can be used to select patients at high risk of VS . The autThe results from this meta-analysis should be evaluated considering some inherent limitations. First of all, we could not assess all the published PTA protocols because the limited information extracted from the included studies did not allow a coherent statistical analysis. Secondly, we could not include randomized controlled trials in this study, and the results are based on observational studies, such as cohort, cross-sectional, or case-control. Thirdly, we did find a high degree of heterogeneity regarding the reporting of sensitivity and specificity data. All these limitations could derive from the disparity of data reported in the literature about the topic. Moreover, it is expected that PTA protocols will be updated based on the data emerging from the new integrative technologies that use artificial intelligence ,43. Further studies, on larger cohorts of patients, or several randomized controlled trials could represent scientific material of higher quality for the next meta-analysis. Meanwhile, we consider that our results support the use of pure-tone audiometry protocols in low resource settings, at least for the risk stratification of patients with asymmetric hearing loss and a high degree of suspicion for vestibular schwannoma. Newer technologies, such as those based on artificial intelligence and machine learning techniques, could help in the process of risk stratification of patients who have a high risk of developing vestibular schwannoma ,45,46,47


Well-tolerated and effective drugs for treating chronic pain conditions are urgently needed. Most chronic pain patients are not effectively relieved from their pain and suffer from debilitating drug side effects. This has not only drastic negative consequences for the patients’ quality of life, but also constitute an enormous burden on society. It is therefore of great interest to explore new potent targets for effective pain treatment with fewer side effects and without addiction liability. A critical component of chronic pain conditions is central sensitization, which involves the reorganization and strengthening of synaptic transmission within nociceptive pathways. Such changes are considered as maladaptive and depend on changes in the surface expression and signaling of AMPA-type glutamate receptors (AMPARs). The PDZ-domain scaffold protein PICK1 binds the AMPARs and has been suggested to play a key role in these maladaptive changes. In the present paper, we review the regulation of AMPARs by PICK1 and its relation to pain pathology. Moreover, we highlight other pain-relevant PICK1 interactions, and we evaluate various compounds that target PICK1 and have been successfully tested in pain models. Finally, we evaluate the potential on-target side effects of interfering with the action of PICK1 action in CNS and beyond. We conclude that PICK1 constitutes a valid drug target for the treatment of inflammatory and neuropathic pain conditions without the side effects and abuse liability associated with current pain medication. Acute pain in response to trauma or inflammation serves an important protective purpose conferring clear adaptive advantages. Chronic pain, on the other hand, has no protective purpose and does not confer any adaptive advantages and, as such, is considered maladaptive. Chronic pain may have both physical and psychological causes. Physical causes can involve musculoskeletal, vascular, neurological and oncological conditions, as well as injury to organs or originate from surgery. Chronic pain can be inflammatory, neuropathic  or a combination of both. Neuropathic pain, in particular, is complex and often resistant to commonly used medication. Chronic pain manifests as spontaneous pain  as well as amplified evoked pain responses to noxious  or non-noxious  stimuli.Chronic pain is one of the ten most prevalent diseases worldwide affecting approximately 20% of the adult population, and the burden caused by chronic pain is escalating ,2,3. TheDespite various etiologies of chronic pain, including the involvement of different neuropathic and inflammatory components, central sensitization, caused by increases in membrane excitability, synaptic efficacy, or a reduced inhibition, has emerged as a critical biological mechanism giving rise to convergence and amplification of nociceptive sensory inputs as well as the spontaneous sensation of pain . At the PICK1 (protein interacting with C kinase 1) is a functional dimer with two PDZ (PSD95/Disc-Large/ZO-1) domains flanking the central, membrane-binding BAR (Bin/amphiphysin/Rvs) domain that also mediates its dimerization . As indiWhile it is well established that PICK1 can bind GluA2 containing, but not GluA2 lacking, AMPA receptors, parallel work has nonetheless suggested a role for PICK1 in regulating the surface expression of GluA2 lacking CP-AMPAR. The first evidence of such a role was obtained in hippocampal slices, demonstrating that viral overexpression of PICK1 increased CP-AMPAR expression . This waMechanistically, we suggest that PICK1 serves (at least) two molecular functions in relation to AMPARs through its PDZ domain interaction with the GluA2 C terminus . The firIt might appear counterintuitive as to how this would affect the synaptic integration of CP-AMPARs, which do not include the GluA2 subunit and, consequently, are not directly associated with PICK1. There are several ways in which this could be envisioned, but the simplest is a phosphorylation-dependent destabilization of GluA2 containing surface receptors , thereby lowering their availability for synaptic integration by a mechanism similar to LTD. This would in turn favor the synaptic integration of GluA2-lacking CP-AMPARs, depending on, e.g., the GluA1 phosphorylation status of these receptors , right. Soon after their discovery, the manner by which PDZ-domain scaffold proteins bind to the GluA2 C terminus affecting the spinal cord AMPAR transmission was investigated. The studies, which were based on the use of inhibitory peptides, pointed to a role of GRIP1 rather than of PICK1, with GRIP1 mediating the recruitment of AMPARs to silent synapses . PICK1 wAnother interesting question is the role of PICK1 in relation to the NMDAR-dependent phosphorylation of GluA2 S880 by PKC, and the subsequent internalization of GluA2 in inflammatory pain . Indeed,Collectively, these studies highlight PICK1 as an attractive target for the treatment of chronic pain conditions by substantiating the critical involvement of PICK1 in the development of neuropathic pain-related mechanical allodynia in particular. Mechanistically, it is plausible that this function of PICK1 is the result of the maladaptive regulation of AMPAR surface expression with the phosphorylation of GluA2 S880 as a reliable proxy. It remains to be shown, however, using, for example, the GluA2 K882A knock-in mice , whetherDespite the convincing evidence for the role of PICK1 in the regulation of AMPAR-dependent plasticity in chronic pain, it is worth considering that PICK1 interacts with many proteins in addition to PKCα and GluA2. Most of these interactions are mediated by the PDZ domain and, hence, all the manipulations to PICK1 mentioned above could be the result of blocking the binding of other interaction partners. To explore the overlap of the PICK1 interactome with the pain-related proteome, we took advantage of the bioinformatic tool StringApp  in CytosFrom the expression pattern of the pain-associated PICK1 interaction partners, we note that the heteromeric BAR domain interaction partner ICA69 is expressed in the DRG and spinal cord, and the relevant supraspinal regions. On the other hand, PKCα is not highly expressed in the DRGs or spinal cord, compared to the supraspinal areas in either mice or humans and, overall, the human expression is relatively low c,d. NotaPICK1 is an untraditional intracellular target, in the sense that it is a scaffold protein without an enzymatic or independent signaling function. Moreover, the PDZ domains, in general, have proven to be challenging to target with sufficient selectivity and affinity. The first efforts to block the PICK1 PDZ domain employed a myristoylated version of the GluA2 C terminus, as previously described ; howeverIn this section, we evaluate the putative risks of PICK1 inhibition based on the available preclinical data from studies on PICK1 KO mice and from human genetic studies. Evidently, this picture could significantly diverge in humans, and the actual risk of a given compound would relate to biodistribution among other things. Nevertheless, our current knowledge would constitute the vantage point of a risk assessment to guide preclinical toxicology in a drug-development process.General excitability: current treatments for chronic pain, including anti-convulsants and opioids, are limited by their relative narrow therapeutic window between efficacious pain relief and side effects, such as drowsiness, apathy, and fatigue, which relate to their on-target actions on Gαi-coupled opioid receptors and the voltage-gated calcium channel, respectively, to reduce synaptic transmissions throughout the CNS. Such side effects would not be anticipated by PICK1 inhibition from known mechanisms of action. Accordingly, PICK1 KO mice exhibit normal locomotor functions, including placing, grasping, and righting reflexes. Additionally, PICK1 KO mice do not display convulsions, hypermobility, spontaneous activity, or other significant difference in their general behaviors [ehaviors ,63,64. Hehaviors . Likewisehaviors . We haveehaviors ,67.Compulsive behavior: addictive side effects of pain-relieving drugs, and, in particular, for opioids, is another key concern. For novel drug targets for chronic pain, it is thus of outmost importance that they do not lead to reinforcing behaviors. In PICK KO mice, we showed that the acute locomotor response to a single injection of cocaine was markedly attenuated. Moreover, in support of the role of PICK1 in neuroadaptive changes induced by cocaine, we observed a diminished self-administration of cocaine in the KO mice [ KO mice , and the KO mice . In acco KO mice . This su KO mice . ConsequMemory function: given the wealth of data involving PICK1 in synaptic plasticity, perhaps the most immediate concern for PICK1 as a drug target is centered around memory performance and flexibility. Nonetheless, the only finding implicating PICK1 in memory function in vivo is a reduced latency in the step-through inhibitory avoidance test observed in adults, but not juvenile PICK1 KO mice [ KO mice . Since t KO mice . AlthougAlzheimer’s Disease: a genetic analysis addressing the association between the PICK1 gene and the risk of Alzheimer’s disease (AD) in the Chinese Han population identified 7 single nucleotide polymorphisms in the PICK1 gene, with rs149474436 being less and rs397780637 being more frequent in AD patients [patients . This stpatients . On the patients , suggestpatients .Schizophrenia and cognition: some studies proposed PICK1 as a schizophrenia susceptibility gene. This association is partly coupled to the PICK1 PDZ domain interaction with serine racemase, an enzyme that converts L-serine to D-serine in glial cells [al cells . Servingal cells . In addial cells ,91, althal cells . It has al cells , but simal cells . UltimatMale infertility: PICK1 homozygous KO males are infertile, whereas heterozygotes demonstrate fertility comparable to wild-type mice [ype mice . Male feype mice . Moreoveype mice . Mechaniype mice . Taken tGrowth retardation and glucose homeostasis: the role of PICK1 in metabolic regulation was first implied by the distinct expression in peptidergic neurons in fruit flies [it flies , and in it flies ,100. Fouit flies . These dit flies ,100 and it flies . Similarit flies , and it it flies . Recentlit flies .Cancer: several studies have observed altered PICK1 expression in human cancers, but the results have been somewhat conflicting [flicting ,108,109.flicting . Additioflicting , presentflicting . On the flicting . Other sflicting . Finallyflicting . Taken tThe research conducted over the last two decades has implicated PICK1 in the maladaptive plasticity of AMPAR, which is critical to central sensitization in chronic pain conditions; however, in the present study, we stress that several other PICK1 interaction partners are implicated in pain. The role of PICK1 in the condition of chronic pain was demonstrated by the use of membrane-permeable PDZ-binding peptides, KO mice, and antisense knock-down strategies; however, the development of small molecules to inhibit the PICK1 PDZ has proven to be a challenge. The recent advent of a high-affinity bivalent PDZ-domain peptide inhibitor may pave the way to the development of a novel efficacious treatment of chronic pain conditions. Current data and analysis of the putative on-target side-effect profile suggest little overlap with the side-effect profile of current pain medication involving addiction, drowsiness, apathy, and fatigue, but warrants a further investigation of the putative effects on memory and cognition, as well as on male fertility and neoplasm. In summary, we believe that PICK1 stands out as a highly promising novel drug target to address the significant unmet need of chronic-pain patients.® filtered using the keyword “Pain” and restricted to genes with a relevance score higher than 5, as well as the String Database (String-db) using the disease query “Pain disorder” [https://www.uniprot.org, accessed on 13 October 2021), resulting in a database containing 1122 proteins. The UniProtKB AC/IDs were used to build a functional protein-association network with the stringApp [p-values using the Benjamini–Hochberg procedure . The FDR threshold was set to 0.01 and terms were filtered for redundancy with a cutoff of 0.5 (default setting in the stringApp), resulting in 26 significant terms from the GO Biological Process, GO Molecular Function, GO Cellular Component, and KEGG Pathways. The terms were manually curated to exclude broad terms and terms with less than 3 associated genes, resulting in 16 terms.For isorder” . All dattringApp  in CytosTechnologies describing tPD5 and related work are disclosed in two patent applications presently being handled by the patent authorities in the E.U. and U.S.A.


The aim of this study is to establish the prevalence, ultrasound manifestations, and signs of thyroid nodules  in the healthy population, in order to provide a basis for the early diagnosis, treatment, and health education for clinical prevention of thyroid diseases.  The data of 2098 physical examination clients who underwent thyroid color Doppler ultrasound examination at our hospital's physical examination center from January 1, 2017, to December 31, 2018, were randomly selected, and the prevalence, size, nature, number, and Thyroid Imaging Reporting and Data System (TI-RADS) classification of the thyroid nodules were statistically analyzed. p < 0.01), and the prevalence increased with age (p < 0.05). The composition ratios of multiple nodules and single nodules in each age group were compared, and the difference was statistically significant (p < 0.05). Detected thyroid nodules were mainly solid, and they were significantly different compared to the cystic and mixed nodules (p < 0.01). The actual classification in our study is consistent with the theory of TI-RADS1∼5 classification.  Of the 2098 results analyzed, 992 cases (47.28%) had thyroid nodules; 327 (15.59%) males and 665 (31.70%) females. The prevalence among females was significantly higher than that among males ( In conclusion, to sum up, thyroid color Doppler ultrasonography is very important for screening thyroid nodules in healthy people during routine physical examination, especially in high-pressure and high-intensity working environment, post-menopausal women, and men with family members with a history of thyroid cancer. Therefore, high-risk groups are encouraged to have regular thyroid ultrasonography, while health care workers are encouraged to publicize the risk factors of thyroid nodules and the importance of routine thyroid ultrasound examination. Today, changes in people's diet structure and the fast-paced nature of work with high pressure have led to a yearly increase in the prevalence of thyroid nodules . ClinicaFeatures of color Doppler ultrasound include on-invasiveness, convenience, and high repeatability and can be used to rapidly acquire the radiographic features within the thyroid lesions. Ultrasonography has become the preferred imaging method for preoperative diagnosis, postoperative follow-up, and screening of thyroid nodules. The improved TI-RADS, when combined with CEUS, could significantly improve the diagnostic accuracy for thyroid nodules, especially for TI-RADS class-4 thyroid nodules. With the increasing popularity of color Doppler ultrasound in routine physical examination, the detection rate of thyroid nodules continues to increase. In order to understand the epidemiological features of thyroid nodules and set a basis for its early diagnosis and prevention, 2098 results of color Doppler ultrasound scans conducted between January 1, 2017, and December 31, 2018, at the physical examination center of our hospital were randomly selected and statistically analyzed. Their epidemiological characteristics and key associations have been summarized and reported as in this paper.From January 1, 2017, to December 31, 2018, a total of 2111 patients were randomly selected for high resolution B-type thyroid ultrasound examination at the physical examination center of our hospital. 13 patients who had previous history of thyroid disease and surgery were excluded, leaving a total of 2098 eligible for the study: 891 males and 1267 females. They had a mean age of 44.02 ± 6.02 years, ranging from 18 to 76 years. Philips color Doppler ultrasound machine was used with the probe frequency adjusted to between 4 and 12 MHz. The patient was positioned in a supine position, the neck assisted with a pillow, and the head tilted back as far as possible to fully expose the thyroid gland. Multiple sections of the thyroid were then scanned, and the size, shape, and number of nodules , internal echo , edge condition, whether there is calcification in the nodule , aspect ratio, etc. were recorded. The signs of thyroid nodules in different patients were analyzed [According to Horvath et al.  Figure , the criχ2) was used to compare the composition ratio of multiple nodules and single nodules in all age groups, with the statistical significance set at P < 0.05. Pathological results were taken as the gold standard, and the percentage of malignancy was calculated according to the TI-RADS classification.Statistical data analysis was conducted using SPSS software, version 21.0. The rate of detection of the thyroid nodules was determined by gender and 6 age groups. The chi-square test .Of the 2098 survey cases, color Doppler ultrasound detected 992 thyroid nodules, with a detection rate of 47.28%. Analyzed by gender, the detection rate was 15.59% in males and 31.70% in females, respectively; the difference between the two is statistically significant .Stratified by age groups, the detection rate among cases in the 18–29 age group was the lowest and gradually increased with age. Among males, the 60–69 age group had the highest rate 65.38%), while for females, the 50–59 age group had the highest rate (76.51%) , and the detection rate gradually increased with increase in age. The composition of the thyroid nodules were mainly solid nodules, which were significantly different from cystic nodules and mixed nodules   had multiple nodules, while 506 (51.01%) had single nodules. There was significant difference in the constituent ratio of multiple nodules and single nodules at different age groups (.01) see .Of the 992 cases with thyroid nodules, 425 cases had TI-RADS class 1 and class 2 ; 336 cases had TI-RADS class 3 ; 107 cases had TI-RADS class 4a ; 91 cases had TI-RADS class 4B ; 24 cases had TI-RADS class 4C ; and 9 cases had TI-RADS class 5 . The actual malignant percentages of TI-RADS class 1 ∼ 5 were 0, 0, 0.6%, 4.67%, 9.89%, 75.00%, and 88.89%, respectively. See D ≥ 0.5 mm and can find thyroid nodules very early, which is of great significance to provide treatment basis for clinicians.The thyroid is an important endocrine organ of human body. Its main function is to regulate human metabolism, psychological development, physical development, and emotional state. Thyroid nodule (TNs) is a frequently occurring disease in the endocrine system. It is a mass in the thyroid tissue caused by abnormal hyperplasia of thyroid glands for various reasons. The prevalence of thyroid nodules in China continues to increase and is attributed to factors such as population aging, great social pressure, emotional internal injury, autoimmune disease, salt iodization, and living environment ; becauseP < 0.01) and when stratified by age groups (P < 0.01). This result is consistent with a similar study by Luo Jingmei [This study discovered that the detection rate of thyroid nodules in randomly selected patients is significantly higher among women (31.70%) compared to men 15.59%), both as a whole , and the proportion of multiple nodules increased with age. In both genders, the nodules were mainly solid (75.60%), which was statistically different from cystic nodules and mixed nodules (P < 0.01). This finding is consistent with the results of two similar studies by Xing et al. [Analyzing by types and composition, our results showed that out of the 992 cases with thyroid nodules, 506 (51.01%) had multiple nodules, while 486 (48.99%) had single nodules. There was indeed a significant difference in the composition ratio of multiple and single nodules at different age groups (g et al.  and Xu eg et al. . HoweverHigh frequency color Doppler ultrasound is currently the first choice for detecting thyroid nodules . It can In conclusion, color Doppler ultrasonography of the thyroid has become the first-choice method for routine physical examination of the thyroid, aiding early diagnosis of thyroid diseases. It is advantageous because the whole process is noninvasive, dynamic, and real-time and has great reproducibility . Women a


Proteobacteria, Acidobacteria, Chloroflexi, Verrucomicrobia, and Planctomycetes were found to increase in abundance from arid to humid climates, while Actinobacteria and Gemmatimonadetes decreased along the transect. Bacterial-community structure varied with climate and aspect and was influenced by pH, bulk density, plant-available phosphorus, clay, and total organic-matter content. Higher bacterial specialization was found in arid and humid climates and on the south-facing slope and was likely promoted by stable microclimatic conditions. The presence of specialists was associated with ecosystem-functional traits, which shifted from pioneers that accumulated organic matter in arid climates to organic decomposers in humid climates. These findings provide new perspectives on how climate and slope aspects influence the composition and functional capabilities of bacteria, with most of these capabilities being involved in pedogenetic processes.Soil bacteria play a fundamental role in pedogenesis. However, knowledge about both the impact of climate and slope aspects on microbial communities and the consequences of these items in pedogenesis is lacking. Therefore, soil-bacterial communities from four sites and two different aspects along the climate gradient of the Chilean Coastal Cordillera were investigated. Using a combination of microbiological and physicochemical methods, soils that developed in arid, semi-arid, mediterranean, and humid climates were analyzed.  Soils play a relevant role both in the functioning of the environment and in life sustainability ,2. Soil Climate is one of the dominant factors that control soil formation and stabilization . ClimatiIn addition to regional climate, aspect is a crucial topographic factor that affects microclimatic conditions through the amount of solar radiation and precipitation received, thereby regulating temperature, evapotranspiration capacity, and water availability in the local soil ,26. ThesThe Chilean Coastal Cordillera provides a large latitudinal extension that covers a wide climate gradient ranging—from north to south—from extremely arid to semi-arid, mediterranean, and cold and humid climates. This gradient is characterized by tremendous diversity of ecological niches and a high level of environmental heterogeneity on the same bedrock . These cIn considering the crucial role of pedogenetic processes in soil formation and ecosystem development, the following question arose: What is the impact of regional climate changes (gradient) and local climate changes (aspect) on bacterial communities that are involved in soil formation and stabilization? With this question as a starting point, the main objective of the present study was to determine changes in the diversity, structure, and degree of specialization of bacterial communities on the north- and south-facing slopes along a climatic gradient on the Chilean Coastal Cordillera in order to better understand the role that these communities play in pedogenetic processes. For this purpose, samples from four different study areas along the Coastal Cordillera—that is, Pan de Azúcar National Park, Santa Gracia Natural Reserve, La Campana National Park, and Nahuelbuta National Park—were used for 16S rRNA high-throughput sequencing, soil physical analyses, and soil chemical analyses. The results highlight the diversity and functional capabilities of soil bacteria, which shifted from pioneers that accumulated organic matter in arid climates to organic decomposers in humid climates.The research was carried out at four primary study areas (PSAs) along the Chilean Coastal Cordillera  within tSoil samples were collected from the northern and southern mid-slope positions  along the Chilean Coastal Cordillera during spring 2016. Sixteen samples per PSA were collected, which represented two slope aspects and four different depth increments with two biological replicates each (with a distance of 50–100 m). The soil-depth increments were 0–5 cm, 5–10 cm, 10–20 cm, and 20–40 cm. The sampling method allowed the analysis of regional variations between PSAs along the climate gradient, as well as the influence of slope aspects on the microbial communities. The soil samples were taken devoid of vegetation in order to reduce the impact of the dominant vegetation on the abundance and diversity of the microbial communities. Samples were sieved at 2 mm and stored at −20 °C for further analysis.2 solution), plant-available phosphorus (plant-available P), bulk density (BD), the carbon:nitrogen ratio (C/N), total nitrogen (Nt), total sulfur (St), and pedogenic oxides in the soil (Feox/Fed)—had a significant effect on the bacterial community and were used for further analysis in the present investigation [A methodology for and results from both soil physical analyses and soil chemical analyses were detailed by Bernhard et al. . From thtigation  (Supplem2 (25 mM), 0.25 µL of BSA (20 mg/mL), and 1 µL of dNTP mix (5 mM). The amplification process was performed in a T100TM Thermal Cycler  with the following cycling program: initial denaturation at 95 °C for 5 min followed by 10 cycles of denaturation at 95 °C for 30 s, annealing for 45 s with decreasing temperatures from 65 to 55 °C (decreasing 1 °C/cycle), elongation at 72 °C for 1 min, 30 cycles of denaturation at 95 °C for 1 min, annealing at 55 °C for 30 s, elongation at 72 °C for 1 min, and finally, an elongation step of 7 min at 72 °C. PCR products were purified using carboxyl-coated magnetic beads  following the manufacturer’s recommendations. All amplification products were pooled equitably in a final concentration of 30 ng of DNA and sequenced together. Sequencing of the pooled samples was performed using HiSeq high-throughput sequencing of bacterial and archaeal 16S rRNA genes and was conducted on an Illumina HiSeq (2 × 250 bp) by Eurofins GATC Biotech .Total genomic DNA was extracted using the PowerMax DNA isolation kit (sixteen samples for AZ) and the PowerSoil DNA isolation kit   according to the manufacturer’s protocol. Each sample consists of three pooled DNA extraction technical replicates and a PCR in duplicate prior to sequencing. The V4 region of the bacterial small-subunit rRNA gene (16S) was PCR-amplified using barcode-tagged multiplex identifiers—described by Caporaso et al. —that corhttp://www.ebi.ac.uk/ena) with BioProject ID PRJEB38745.Raw data quality was checked using FastQC . Dual inStatistical analyses were performed using 16 samples per site that corresponded to two aspects, two biological replicates, and four depths. All samples were subsampled at 32,000 reads and transformed into relative abundances in order to standardize the data. Biological duplicates were analyzed separately. The physicochemical data were standardized by subtracting the mean and dividing by the standard deviation.p-level of significance: <0.05) with 999 permutations using the Adonis function. An indicator-value analysis  was performed that avoided rare taxa (>0.1%) when analyzing habitat specialization. IndVal analysis is based on an ASV’s fidelity, and the relative abundance within a particular PSA is calculated using the “indval” function in the labdsv package [p < 0.05) were considered as a good indicator of specialization for either PSA. In contrast, ASVs with high incidence (>75% of the samples) and high relative abundance over all sites (>0.1%) were considered arbitrarily as habitat generalists. The taxonomic relative abundances across samples were visualized through bubble plots using the package ggplot2 [p < 0.05) using CANOCO 5.0 software  [Diversity- and community-structure analyses were performed using R studio version 3.6.1. The defined ASVs were used to calculate alpha-diversity indices using the vegan package , which i package . Only AS ggplot2 . FinallyNY, USA) .t content decreased, while Feox/Fed, Nt, and the C/N ratios increased from north to south. The plant-available P increased from AZ to SG and then decreased to NA. Furthermore, plant-available P and Feox/Fed showed a significantly higher SFS value than did the NFS in LC.The four locations along the transect—that is, the arid climate of AZ, the semi-arid climate of SG, the mediterranean climate of LC, and the humid climate of NA—revealed trends among sampling locations of ten physicochemical parameters . In summHigh-throughput sequencing resulted in a total of 18.4 million raw reads from all 64 soil samples. After filtering, merging, and removing chimeras, a final number of 14.9 million reads—corresponding to 81.1% of the total—was obtained . Of thesp < 0.05). For the SFS, pH (R = −0.63), St (R = −0.47), and Nt (R = 0.43) were correlated with the Shannon index.Alpha-diversity values were calculated from subsampled ASVs and visualized using R . For botActinobacteria (30.2%), Proteobacteria (21%), Acidobacteria (12.9%), Chloroflexi (9.74%), Verrucomicrobia (6.5%), Planctomycetes (6.1%), and Gemmatimonadetes (4.4%) , Proteobacteria (15.3%), and Chloroflexi (8.8%). In SG and LC, the three dominant phyla were Actinobacteria , Proteobacteria , and Acidobacteria . Finally, NA was dominated by Proteobacteria (29.1%), followed by Acidobacteria (18.5%) and Chloroflexi (16.5%).Along the climate gradient, seven phyla dominated in both slope aspects: s (4.4%) . These sobserved . The mosp < 0.01), while the NFS and the SFS were significantly different only in the southernmost sites of LC and NA (p < 0.05).PERMANOVA was used to explain differences in bacterial-community structure along the climate gradient and the different slope aspects . The fout, and lower levels of weathering, while NA had a positive relation with Feox/Fed. In the SFS, the explanatory variables accounted for 30% of the total variance of the bacterial distribution, with CCA 1 and 2 explaining 35.9% and 27.1% of this total variance, respectively. The graph reveals that NA was influenced positively by clay content and was influenced negatively by pH and BD.The relation between ASVs and physicochemical parameters was identified using CCA a,b. ResuActinobacteria, Proteobacteria, Acidobacteria, Chloroflexi, and Verrucomicrobia, which together represent thirteen different taxa , the Blastococcus genus, and the JG30-KF-CM45 family displayed a predominant abundance in SG and LC. ASVs from the Solirubrobacteraceae family, the Xanthobacteriaceae order, the Rhizobacter order, and the Elsterales class were more abundant in NA. In contrast, the Ralstonia genus predominated in AZ. Regarding depth, ASVs from the Rubrobacter genus and the JG30-KF-CM45 family were more abundant on the surface and decreased with depth.Of the 39,195 ASVs, only 301 and 302 had a mean of over 0.1% for the NFS and the SFS, respectively. Of these, 16 and 7 were generalists for the NFS and the SFS, respectively, despite the variability of the ecosystems. The generalist community in the NFS was composed of five phyla: ent taxa a. On theent taxa b. The dit. In contrast, NA was positively influenced by clay and TOC. Considering the SFS , LC (the 67-14 family), and NA (the IMCC26256 order and the Conexibacter genus). The family Thermomonosporaeae was a prolific specialist in the SFS of AZ, whereas in the NFS, this family was a low-abundant unspecialized ASV. Other specialists were found in both slope aspects, which occupied different PSAs. An example is the Gaiellales order, which is a specialist on the NFS of SG and the SFS of LC. The same is true for the Acidomicrobiia class, which is a specialist on the NFS of SG and on the SFS of NA.Acidobacteria  were frequently NA specialists in both aspects. Other ASVs from the Subgroup 6 class and the RB41 genus were LC specialists. Furthermore, many specialist ASVs were abundant in one of the two aspects. Members of the Subgroup 7 order were specialists in the NFS of AZ and NA, while the Bryobacter and Granulicella genera were specialists in the SFS of NA. On the other hand, in Proteobacteria, ASVs from the Elsterales and WD260 orders, the A21b and Xanthobacteriaceae families, and the Acidibacter genus were NA specialists, while ASVs from the Sphingomonas genus were AZ specialists.AD3 and TK10 classes and the HSB OF53-F07 genus that belonged to the Chloroflexi phyla were NA specialists, while ASVs from the C0119 order were SG specialists. Firmicutes  and Gemmatimonadetes  were AZ specialists. The family Gemmataceae of the Planctomycetes phylum were NA specialists. Finally, ASVs from the Candidatus Xiphinematobacter genus that belonged to Verrucomicrobia phyla were specialists in NA.Regarding phyla with a low number of specialists, ASVs from the ox/Fed and silt, while SG and LC were influenced by plant-available P, and AZ was influenced by pH. For the SFS, 61% of the total variance was explained by physicochemical parameters , clay content, microaggregates (<20 µm), recent weathering processes (>Feox/Fed), and lower plant-available P (efficient P acquisition), which is consistent with the preference of Actinobacteria and Proteobacteria in NA [The microbial-community structure at each site is significantly different from that at the other sites, which highlights the effect of regional climate along the gradient. From arid to humid climates, the microbial-community structure changes according to soil physical properties and soil chemical properties, as already discussed for pH. In addition to the pH, BD, plant-available P, Ferocesses . For insa Desert ,34,57. Aormation ,11,58. Iia in NA ,59,60,61Actinobacteria with a potential role in nitrogen metabolism—such as Blastococcus, Rubrobacter, and Solirubrobacteraceae—are classified as generalists in different ecosystem types, which include deserts, shrubs, and tree groves [Candidatus Udaeobacter genus of Verrucomicrobia belongs to the most abundant soil bacteria worldwide and has a high capacity to produce antibiotics that allow its members to compete efficiently for space and nutrients  in soils [Proteobacteria, Bradyrhizobium and Sphingomonas have been described as some of the most abundant and ubiquitous genera on six continents due to their multiple survival strategies in soils [Rhizobiales (Bradyrhizobium and Xanthobacteriaceae) can fix nitrogen in symbiotic association with plants or under free-living conditions due to the presence of the nifH gene [Microorganisms exist as habitat generalists or specialists in response to the climate gradient and to environmental filtering . Generale groves . The Canin soils . Within in soils ,67. For ifH gene ,69,70. TifH gene . Since gifH gene ,71.Pelomonas and Ralstonia [Anoxybacillus, Staphylococcus, and Paenibacillus (of the Bacillus order) are adapted to the use of recalcitrant carbon substrates and inorganic nutrients, thereby allowing for the accumulation of organic matter [Conexibacter and 67-14  have been linked to chemosynthetic CO2 fixation and ferrous–ferric redox reactions, thereby enhancing weathering and the availability of micronutrients in soils [Sphingomonadacea family improves soil nutrition by degrading polycyclic aromatic hydrocarbons with hydrophobic properties [Gemmatimonadetes that are adapted to low soil moisture have been associated with phosphorous metabolism [In addition to the study of generalists, we also examined specialists. AZ harbors a large community of specialists, which are highly adapted bacteria that can survive the extreme environmental conditions of the desert, including the nearly complete lack of organic matter ,36. The alstonia . Anoxybac matter ,68,69. Cin soils ,72,73. Toperties . Gemmatitabolism ,14,74. Atabolism ,10. Thistabolism ,76.ox/Fed in the soil. The increased organic matter in this site satisfies the nutrient demand through efficient nutrient recycling via microorganisms [Acidobacteria phylum can hydrolyze polymers such as cellulose, hemicellulose, or starch due to the high content of glycoside hydrolases [Subgroups 2 and 6, Candidatus Solibacter, Acidobacteriales, and Bryobacter have been documented in global acidic forest soils [Proteobacteria, species of the Burkholderiales order are efficient mineral-weathering bacteria and are also involved in the decomposition of lignocellulose due to the presence of abundant laccase genes [Proteobacteria, such as WD260, A21b, and Acidibacter, are associated with the decomposition of C-labile compounds, such as glucose and naphthalene [Alphaproteobacteria and Solibacterales have abundant genes for phosphatase-enzyme synthesis and phosphorous solubilization, which can mobilize phosphorus in soils with a low P content [Burkholderia-Caballeronia, Variovorax, Xanthobacteraceae, Granulicella, HSBOF53-F07 (the genus belonging to Chloroflexi), JG30a-KF-32 (the family belonging to Acidobacteria), and Gemmataceae [Unexpectedly, NA contains the most significant number of specialists linked to acidification and increased TOC, clay, and Ferganisms ,78. Our drolases . From thst soils ,81,82. Fse genes ,82. In ththalene . Alphapr content . Finallymataceae ,85. The AZ and NA are unique hotspots of bacterial specialization along the climate gradient, which suggests that they represent climatically and topographically stable ecosystems ,87. SpecOverall, our findings indicate that regional climate is a powerful factor in shaping bacterial communities along the Chilean Coastal Cordillera. We suggest that SG and LC are topographically and climatically unstable ecosystems that promote a low number of bacterial specialists and high bacterial diversity in order to cope with environmental disturbances. On the other hand, the ecosystem stability in AZ and NA has allowed the bacterial community there to specialize in the use of specific resources with strong ecological preferences that are linked to the bacteria’s role in both soil formation and ecosystem functioning .Thermomonosporaceae family reaches 49.9% abundance in the SFS compared with 1.4% abundance in the NFS, which indicates a high degree of adaptation to specific microclimatic conditions. In addition to radiation and moisture, the occurrence of BSC cover and the thickness of this cover are important factors that control slope-scale bacterial diversity in arid sites by influencing moisture and physicochemical properties [The slope aspect causes changes in both the diversity and structure of bacterial communities that result from particular microclimatic and physicochemical differences ,28. The operties ,97. Sincoperties . Overalloperties .ox/Fed [The bacterial-community structure of the NFS only differs significantly from that of the SFS in the two southernmost sites of LC and NA, which is mediated by aspect-induced changes in soil chemical properties. This finding suggests that the effect of the slope aspect depends on climate and latitude. Latitude accounts for the increasing differences in radiation time between the SFS and the NFS when moving southward and alters aspect-related soil pH, plant-available P, clay content, BD, and Feox/Fed . Of thesox/Fed . Moreoveox/Fed . We suggox/Fed ,99.In general, along the gradient, a higher number of generalists was found in the NFS, and a higher number of specialists was found in the SFS. These findings suggest that more stable conditions exist in the SFS than in the NFS. This suggestion is supported by the findings of Pen-Mouratov et al.  in the “Our findings reveal that soil-bacterial diversity and changes to the microbial-community structure depend on local filters that are promoted by the slope aspect. However, the response is not constant along the climate gradient. We suggest that the response of bacterial communities could be considered a result of the interplay between different factors, such as microclimate conditions, soil cover (BSC and/or vegetation), and physicochemical properties, all of which depend primarily on large-scale factors, such as climate and latitude. These findings highlight the importance of the remarkable and unique climate gradient of the Chilean Coastal Cordillera in discriminating among multiple factors that shape microbial communities. Moreover, we encourage similar approaches be taken to predict the function and impact of microbial communities in ecosystem development.Our study revealed shifts in the diversity and structure of microbial communities that were induced by climate and aspects along the Chilean Coastal Cordillera, and a correlation between these shifts and prominent indicators of soil formation was found. Along the studied climate gradient, the arid and humid sites behave as stable ecosystems that promote low diversity and high bacterial specialization. Moreover, the south-facing slope represents a more stable ecosystem than the north-facing slope and hosts a greater number of specialists. The presence of bacterial specialists is associated with ecosystem-functional traits. In the arid climate, bacterial specialists are mainly dominated by pioneer microorganisms that are involved in organic-matter accumulation, which is key in the initial pedogenetic process and in ecosystem development. In contrast, the humid-climate bacteria specialists are dominated by microorganisms that are involved in recycling organic compounds as a source of plant nutrients and soil stabilization. Nonetheless, soil-manipulation experiments coupled with transcriptomic analysis would be required in future studies in order to determine whether these bacteria are actively accomplishing this role. In the case of the semi-arid and mediterranean sites, we observed a low number of specialists and a high level of bacterial diversity for coping with the climatic and topographic variability. Our findings provide a basis for future studies on stabilizing, maintaining, and improving different soil ecosystems that are exposed to climate change.


To investigate the influence of HbA1c level and GWG on pregnancy outcomes in pregnant women with GDM.A total of 2,171 pregnant women with GDM were retrospectively included and categorized as follows: (1) normal (HbA1c <6%) and elevated (HbA1c ≥6%) HbA1c groups according to the HbA1c level in the second trimester, and (2) inadequate, appropriate, and excessive GWG groups according to the IOM guidelines.P < 0.05). The risks of primary cesarean section, hypertensive disorders during pregnancy, and macrosomia in pregnant women with excessive GWG were significantly higher than those with inadequate and appropriate GWG (P < 0.05). When GWG was appropriate, the risk of hypertensive disorders during pregnancy in the elevated HbA1c group was higher than that in the normal HbA1c group. When GWG was excessive, the risks of postpartum hemorrhage, macrosomia, and neonatal asphyxia in the elevated HbA1c group were significantly higher than in the normal HbA1c group (P < 0.05).In pregnant women with GDM, advanced age and high pre-pregnancy BMI were high-risk factors for elevated HbA1c. Pregnant women with elevated HbA1c had higher OGTT levels than those with normal HbA1c, and the risks of adverse pregnancy outcomes were higher (Monitoring and controlling blood glucose levels have shown effectiveness in reducing the adverse pregnancy outcomes in women with GDM, particularly for those who had excessive GWG. Gestational diabetes mellitus (GDM) refers to diabetes that appears or is diagnosed during pregnancy in women with normal glucose metabolism or potentially impaired glucose tolerance before pregnancy . With thGlycated hemoglobin (HbA1c) reflects average blood glucose levels during the preceding 8–12 weeks. As the gold standard for assessing long-term glycemic control, it has been widely used in blood glucose monitoring and management of people with diabetes. The increase in HbA1c levels is closely related to the adverse pregnancy outcome . In 2009This study retrospectively included 2,171 pregnant women with GDM, and aimed at exploring the effects of second-trimester HbA1c levels on pregnancy outcomes of GDM and analyzing the role of GWG on the pregnancy outcomes with different HbA1c levels, thus providing the basis for precise intervention and personalized, risk-stratified weight management for gestational diabetic women with different HbA1c levels.A retrospective collection of pregnant women with GDM who had undergone regular antenatal checkups and gave birth at the Women's Hospital School of Medicine Zhejiang University from 1 July 2017 to 30 June 30 2018. Inclusion criteria were as follows: (1) diagnosis of GDM in the second trimester using the oral glucose tolerance test (OGTT); (2) singleton pregnancy; (3) gestational age at birth ≥28 weeks; and (4) complete medical records. Exclusion criteria were as follows: (1) coupled with pre-gestational diabetes; (2) coupled with chronic hypertension, or diseases of the liver, kidney, heart, lung, and other major organs, or coupled with tumors; (3) coupled with Sjogren's syndrome, anticardiolipin syndrome, or other autoimmune diseases. A total of 2,352 pregnant women met the inclusion criteria. After the application of the exclusion criteria, a total of 2,171 cases were eventually included in this study .The relevant information regarding the pregnant women was retrieved through the electronic medical record system of the Women's Hospital School of Medicine, Zhejiang University, which included pregnant women's age, height, pre-pregnancy weight (within 1 month before pregnancy), gestational weight gain, pregnancy parity, OGTT value , second-trimester glycated hemoglobin level, mode of delivery, gestational age at birth, birth weight of newborn, and pregnancy complications, such as hypertensive disorders during pregnancy , placental abruption, postpartum hemorrhage, intrahepatic cholestasis of pregnancy, neonatal asphyxia, etc.The study was approved by the Human Ethics Committee at Women's Hospital, School of Medicine, Zhejiang University.2). GWG (kg) is the difference between the weight before delivery and the weight before pregnancy. According to the 2009 IOM guidelines in the United States, the suggested GWG is 12.7–18.1 kg, 11.3–15.8 kg, 6.8–11.3 kg, and 5.0–9.1 kg for pregnant women with pre-pregnancy BMI categorized as underweight (BMI < 18.5 kg/m2), normal weight (BMI: 18.5 kg/m2−24.9 kg/m2), overweight (BMI: 25.0 kg/m2−29.9 kg/m2), and obese (BMI ≥ 30.0 kg/m2), respectively. One is considered to have inadequate GWG or excessive GWG if the GWG is less than or exceeding the appropriate value, respectively.The normal glucose values of fasting, 1-h and 2-h 75-g OGTT, according to IADPSG criteria, were <5.1 mmol/ L, 10.0 mmol/L, and 8.5 mmol/L, respectively . Any abnGestational hypertension is the first incidence of high blood pressure at ≥20 weeks of gestation, with a systolic blood pressure of 140 mmHg and/or diastolic blood pressure of 90 mmHg in the absence of proteinuria or new signs of end-organ dysfunction. Preeclampsia is defined as a combination of one or more of the following conditions based on the diagnosis of gestational hypertension: proteinuria or other target organ dysfunction or uteroplacental function obstacle. Meanwhile, eclampsia is a seizure that occurs as a result of preeclampsia and cannot be explained by other factors. Placental abruption occurs when the placenta in its usual location becomes partly or totally separated from the uterine wall before the fetus is born after 20 weeks of pregnancy or during labor. Postpartum hemorrhage is defined as bleeding more than 500 ml after vaginal birth or more than 1,000 ml after cesarean delivery within 24 h of the birth of the fetus. Intrahepatic cholestasis of pregnancy is a pregnancy problem that manifests clinically as skin pruritus and increased bile acids in the second and third trimesters. The primary cesarean section is defined as the section that is performed on women who never had a cesarean delivery before. The neonatal Apgar score is presently used to diagnose newborn asphyxia, which is based on five indicators measured at 1 min after birth: heart rate, breathing, muscular tone, laryngeal reflex, and skin color. The normal score is 8–10 points, moderate asphyxia is given 4–7 points, and severe asphyxia is given 0–3 points. Macrosomia is defined as newborns with a birth weight ≥4,000 g.t-test. Count data were expressed in frequency and rate, and the difference between groups was compared using the chi-squared (χ2) test. Multivariate logistic regression analysis was performed to evaluate the effect of glycated hemoglobin and GWG on pregnancy outcomes in pregnant women with GDM, while Fisher's exact test was used to examine the effect of GWG on the pregnancy outcomes of groups with different levels of HbA1c. A p-value of 0.05 or less was considered to be statistically significant. All statistical analyses were done using SPSS Statistics 25.0. Power analysis for total population and high HbA1c group was shown in The data of the normal distribution were expressed as x ± s, and the difference between the means of the two groups was analyzed by the P = 0.031, P = 0.000, P = 0.000, P = 0.016, P = 0.004, respectively). Meanwhile, the risk of hypertensive disorders during pregnancy, postpartum hemorrhage, placental abruption, neonatal asphyxia, and macrosomia was also significantly higher in the elevated HbA1c group   .P = 0.000)  .P < 0.05)  .P = 0.032). In addition, in the excessive GWG group, the incidence of postpartum hemorrhage, neonatal asphyxia, and macrosomia was also significantly higher in the elevated HbA1c subgroup than in the normal HbA1c subgroup   .The second-trimester HbA1c value ≥6% and gestational weight gain exceeding the IOM recommendations were risk factors for adverse pregnancy outcomes in pregnant women with gestational diabetes. In this study, we noted a significantly increased risk of adverse pregnancy outcomes among pregnant women with GDM in the elevated HbA1c group and excessive GWG group compared to those in the normal HbA1c group and appropriate and inadequate GWG groups, respectively. Moreover, GDM women with both high HbA1c levels and excessive GWG had a higher risk of postpartum hemorrhage, neonatal asphyxia, and macrosomia.Gestational diabetes is one of the most common complications of pregnancy . In addiAlthough OGTT is the gold standard for the effective diagnosis of gestational diabetes at present, it is susceptible to short-term lifestyle changes, such as diet and activity before the test. HbA1c is a class of stable compounds in which glucose is covalently bonded to the N-terminal valine residues of the β-chain of hemoglobin . It can In pace with the improvement of living standards and attaching importance to nutrition during pregnancy, recently, researchers have been paying more attention to the appropriate GWG in perinatal health care. For instance, the 2009 IOM revised guidelines for gestational weight gain provide clinicians with a practical basis. The guidelines recommend an optimal amount of GWG for pregnant women according to pre-pregnancy BMI  . AlthougIn general, weight management during pregnancy plays an important role in reducing adverse pregnancy outcomes. However, a few studies were conducted at home and abroad to determine whether weight management during pregnancy can still work for gestational diabetic women with elevated HbA1c levels. Lise L. Kurtzhals et al.  revealedHowever, our study has some limitations. First, given that it is a retrospective study, selection and recall bias was inevitable. Second, the population included in this study was pregnant women attending the Women's Hospital School of Medicine, Zhejiang University, and the sample suggests a lack of representativeness. In addition, the sample size was inadequate, as there were only 49 pregnant women with GDM in the elevated HbA1c group. Moreover, the lack of data on HbA1c levels in the third trimester of pregnancy made it impossible to study the effect of gestational weight gain on this parameter. The above-mentioned deficiencies need to be addressed in further follow-up research.In summary, the second-trimester HbA1c level is an important indicator for predicting adverse pregnancy outcomes in pregnant women with gestational diabetes, and therefore should be taken into account in the regular obstetric examination. Meanwhile, weight management during pregnancy significantly improves pregnancy outcomes, and reasonable weight gain suggestions should be given to the pregnant women in the first trimester of pregnancy based on pre-pregnancy BMI. For pregnant women with GDM who gained excessive weight during pregnancy, it is particularly necessary to conduct obstetric examination and follow-up education, monitor and control blood glucose levels, and strictly manage the weight during pregnancy, in order to further reduce the occurrence of adverse pregnancy outcomes.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by Human Ethics Committee at Women's Hospital, School of Medicine, Zhejiang University. The patients/participants provided their written informed consent to participate in this study.Conceptualization of the study is done by QZ and CL. Data curation is performed by QZ and LZ. Formal analysis and investigated the study are made by QZ, CL, LZ, and QW. QZ contributed to funding acquisition, project administration, and methodology. CL and QW contributed to resources. YC, DC, and LQ supervised the study. QZ, CL, LZ, YC, DC, and LQ validated the study and wrote the original draft. CL performed visualization. QZ, CL, and ZL contributed to writing, reviewing, and editing the manuscript. All authors contributed to the article and approved the submitted version.This work was supported by grants from the National Natural Science Foundation of China (81974234).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.


Vibrio, and promoted the abundance of beneficial bacterium Bacillus. In conclusion, dietary GML significantly reduced the oxidative damage caused by sudden changes in salinity, improved the antioxidant capacity, and alleviated the intestinal flora imbalance in juvenile grouper.Groupers with an initial body weight of 9.10 ± 0.03 g were selected to investigate whether dietary addition of 0 (G0) and 1800 mg/kg glycerol monolaurate  could alleviate the oxidative stress response and intestinal flora imbalance after 0, 6, 12, and 24 h of salinity change in grouper. Experimental results show that the dietary addition of GML significantly reduced the liver MDA content and increased the SOD activity of grouper. The gene expression of CAT and SOD increased and then decreased with time after adding 1800 mg/kg GML, and the highest values were significantly higher than those of the control group. Salinity change had a slight effect on the top four intestinal flora composition of grouper at 0, 12, and 24 h, with changes occurring only at 6 h when Cyanobacteria replaced Actinobacteria. The addition of dietary GML slowed down the intestinal flora disorder, inhibited the colonization of harmful bacterium  With the development of intensive aquaculture, aquatic animals are subjected to increasing stress factors, such as excessive density, water pollution, shock, and sudden changes in salinity and temperature. Environmental stresses may induce excessive production or accumulation of reactive oxygen radicals (ROS) in aquatic animals. They could affect the antioxidant protection capacity of aquatic animals, thus causing oxidative stress in fish and even leading to various disease outbreaks ,2,3.Previous studies have shown that a variety of essential nutrients  and additives  could modulate the immune system of aquatic animals and alleviate the stress experienced by them in the environment. This modulation is one of the effective means of enhancing stress resistance, and it could be a stress management strategy to improve the overall performance of aquatic animals ,5,6. FunDanio rerio [Larimichthys crocea [Lateolabrax maculatus [Litopenaeus vannamei [Pelodiscus sinensis [Glycerol monolaurate (GML) is a natural compound composed of glycerol and lauric acid, a derivative of fatty acids, with remarkable antibacterial, antiviral, and anti-inflammatory properties . Althougio rerio , Larimics crocea , Lateolaaculatus , Litopenvannamei , and Pelsinensis . The aboEpinephelus fuscoguttatus♀ × Epinephelus lanceolatus♂) has become one of the most important breeding species along the southeast coast of China because of its fast growth rate, high nutritional value, and disease resistance [Hybrid grouper  to the basal diet (G0) . Firstly3) for two weeks before the start of the experiment to adapt to the new environment and meet the experimental requirements. Feeding was stopped 24 h before the start of the experiment t [Juvenile groupers were sourced from the Yongsheng Fish Hatchery . All groupers were placed in many fiberglass tanks . Livers were extracted for antioxidant index and gene expression, and intestines were taken for intestinal flora determination.The content of malondialdehyde (MDA) and activity of superoxide dismutase (SOD) were determined by commercial ELISA kits .® Green Pro Taq HS Premix II kit (Accurate Biotechnology (Hunan) Co., Ltd., Hunan, China). Primer sequences were designed using Primer 5 following the cDNA sequences found in NCBI , and the RNA concentration was determined by nuclear acid protein analyzer . Complementary DNA (cDNA) was synthesized by Evo M-MLV RT kit (Accurate Biotechnology (Hunan) Co., Ltd., Hunan, China). All gene expressions of interleukin-6 (IL6), catalase (CAT) and SOD were performed on Roche Light Cycler 480II (Switzerland) using an SYBR in NCBI . The resT method .The DNA from intestinal sample was extracted by Guangzhou Gene Denovo Ltd. using the E.Z.N.A fecal DNA Kit .After extraction of genomic DNA from the samples, the V3 + V4 region of the 16S rDNA was amplified using specific primers with barcodes. The primer sequences were: 341F: CCTACGGGGNGGCWGCAG; 806R: GGACTACHVGGGTATCTAAT. PCR reaction conditions were set according to the description of the previous study.PCR products were first examined by 2% agarose gel electrophoresis then purified with AxyPrep DNA Gel Extraction Kit  following the instructions. The purified amplification products were ligated to sequencing ligations, and sequencing libraries were constructed and sequenced on Illumina.After sequencing the raw reads, the low-quality reads and tags were first filtered using the Usearch software, and the resulting data are called Clean Tag. Secondly, clustering was conducted by applying the Usearch software to access OTU abundance and OTU representative sequences according to Clean Tag. Finally, functional predictions such as species composition analysis, indicator species analysis, and alpha diversity analysis were performed on the basis of OTU sequences and abundance data.p < 0.05), Duncan’s multiple comparisons were adopted. The experimental data were expressed as mean ± standard deviation (X ± SD).Before the application of ANOVA, the homogeneity of variance was analyzed. Data from this experiment were statistically analyzed using SPSS 17.0 software. One-way ANOVA was used to analyze the data, and if there were significant differences (p < 0.05). The MDA content of the G0 and G1800 groups as a whole increased first and then decreased, and it had the highest value at 12 h. As shown in p < 0.05).As seen from p < 0.05, p < 0.05, p < 0.05) and was significantly lower than that of the G0 group at 0, 12, and 24 h  with increasing time throughout the salinity change period. At 0 h, the abundance in the G0 group was significantly higher than that in the G1800 group. At 6, 12, and 24 h, the abundance in the G1800 group was significantly higher than that in the G0 group . The Vibrio in the G1800 group was significantly lower than that in the G0 group, except for 24 h (p < 0.05). The relative abundance of Bacillus in the G0 group increased with time and exhibited the highest value at 12 h (p < 0.05). Meanwhile, the Bacillus in the G1800 group showed no significant difference at 0, 6, and 24 h, but it increased significantly at 12 h (p < 0.05).The relative abundance of Eriocheir sinensis significantly increased the SOD activity and decreased the MDA content in the hepatopancreas [Litopenaeus vannamei. In the present study, compared with the control group, supplementation of 1800 mg/kg GML significantly increased the SOD activity of grouper, improved the antioxidant capacity, and enhanced the ability to remove ROS, while significantly reducing the MDA content, decreasing lipid peroxidation, alleviating oxidative stress, and reducing damage in grouper.The antioxidant system in aquatic animals consists of an enzymatic system (SOD and CAT) and a non-enzymatic system (oxidation products such as MDA and other non-enzymatic antioxidants). SOD is the first line of defense against oxidative stress, scanning excess oxygen radicals (ROS), and increasing the antioxidant level in fish, which, in turn, protects cells from ROS damage . ROS coupancreas . Wang etpancreas  experimeLitopenaeus vannamei. In the salinity stress experiment, the expression levels of CAT and SOD in the G1800 group increased and then decreased, and they had the highest values at 6 and 12 h, respectively, both of which were significantly higher than those in the G0 group. This finding indicated that the peroxidation level was alleviated by the addition of GML, which regulated the delicate balance between oxidants and antioxidants, enhanced the antioxidant capacity of grouper, and reduced oxidative stress. IL6 is one of a family of pro-inflammatory factors that could significantly promote pathological inflammation [Danio rerio diets [CAT belongs to one of the antioxidant enzymes, which could coordinate with SOD to scavenge ROS and prevent or repair oxidative damage . Wang etammation . Studiesio diets . Zhao etio diets  showed tio diets ,26. Inflio diets ,28. The Photobacterium species are pathogenic in a wide range of marine animals, including fish, crustaceans, and mollusks [Photobacterium is a Gram-negative bacterium that prefers salinity environments of 2–4%, and it could produce large amounts of histamine in high-salinity environments [Photobacterium was significantly reduced by the addition of GML. The sudden drop in salinity caused the death of Photobacterium by water uptake rupture in the G0 and G1800 groups, but the death rate in the G1800 group was significantly lower than that in the G0 group. The reason may be that lauric acid (LA), the hydrolysis product of GML, could enter the liver directly through the portal system in free form to participate in organism metabolism. In the absence of apolipoproteins, LA could also enter the mitochondria for oxidation and rapid energy production for the organism [Photobacterium abundance. Vibrio is a salt-loving bacteria, a common pathogen of aquatic creatures, and known as the “scourge” of marine fish [Vibrio [Vibrio could be as low as 20–30 min, thus allowing a rapid response to changing environmental conditions [Vibrio of γ-Proteobacteria at 24%. The relative abundance of Vibrio vulnificus was generally negatively correlated with salinity, and a high abundance of it was observed at 5–10‰ salinity, regardless of temperature [Vibrio abundance. The abundance of Vibrio in the G0 and G1800 groups increased and then decreased over time after salinity change, but the Vibrio abundance in the G1800 group was consistently significantly lower than that in the G10 group. This finding indicated that GML significantly inhibited the multiplication of pathogenic V. vulnificus after a sudden drop in salinity and improved the stress resistance of grouper. Bacillus is widely used as a probiotic in aquaculture. Many experiments have shown that Bacillus could enhance the activity of antioxidant enzymes, increase the expression of immune-related genes and stress-resistant-related genes, and improve the ability of fish to resist pathogenic microorganisms [Bacillus. After 12 h, the abundance of Bacillus in the G1800 group was significantly higher than that in the G0 group. It slowed down the changes in intestinal flora caused by sudden changes in salinity, increased the abundance of beneficial bacteria, and improved the ability of the organism to resist stress response.The intestinal flora is a complex ecosystem with dynamic diversity. A close relationship exists between the intestinal flora and the immune system, playing an important role in maintaining normal intestinal function and the immune function of the body ,30. Dietmollusks . Photobaronments . Long-teronments ,36. The organism . Osmoregine fish . The red [Vibrio . The gennditions . Oberbecnditions  showed tperature . The addrganisms ,44,45,46The supplementation of GML in grouper diet significantly reduced the oxidative damage represented by salinity stress, improved antioxidant capacity, slowed down intestinal flora imbalance, promoted the abundance of beneficial bacteria, and enhanced resistance to oxidative stress. The findings of the present study can add value to the current existing knowledge of beneficial effects of GML in aquaculture. We will make more efforts to produce more research results of GML in aquatic animals.


Individuals with metabolic risks are at high risk of cognitive impairment. We aimed to investigate whether the Thai Cardiovascular Risk (TCVR) score can be used to predict mild cognitive impairment (MCI) in Thai adults with metabolic risks. The study was conducted using secondary data of patients with metabolic risks from Maharaj Nakorn Chiang Mai Hospital. MCI was indicated by an MoCA score of less than 25. Six different TCVR models were used with various combinations of ten different variables for predicting the risk of MCI. The area under the receiver operator characteristic curve (AuROC) and Hosmer–Lemeshow goodness of fit tests were used for determining discriminative performance and model calibration. The sensitivity of the discriminative performance was further evaluated by stratifying by age and gender. From a total of 421 participants, 348 participants had MCI. All six TCVR models showed a similar AuROC, varying between 0.58 and 0.61. The anthropometric-based model showed the best risk prediction performance in the older age group (AuROC 0.69). The laboratory-based model provided the highest discriminative performance for the younger age group (AuROC 0.60). There is potential for the development of an MCI risk model based on values from routine cardiovascular risk assessments among patients with metabolic risks. Individuals with metabolic disease are not only at high risk of cardiovascular events but also cognitive decline. The significant risk factors are also components of metabolic risk, including high blood pressure, elevated blood sugar, elevated waist circumference (WC), and abnormality of plasma cholesterol or triglyceride levels ,2. Many In addition, a significant number of individuals with metabolic diseases also experience cognitive deterioration ,7,8. TheAlthough there is a link between the risk factors for major cardiovascular events and cognitive impairment, screening for MCI in people with metabolic diseases is less common than performing cardiovascular risk assessment . StandarThis study utilized data collected from a previous study examining the possible link between fibroblast growth factor 21 and cognitive decline among patients with metabolic risk factors . AdditioMoCA is the standard tool for screening patients who have MCI. It is a 30-question test. The MoCA is recommended by the Canadian Consensus Conference for Diagnosis and Treatment of Dementia Guidelines for Alzheimer’s disease, and the National Institutes of Health and Canadian Stroke Consortium for Vascular Cognitive Impairment and is available in more than 200 countries around the world . It is u(1)Age, gender, DM, smoking status, SBP, and WC;(2)Age, gender, DM, smoking status, SBP, WC, and height;(3)Age, gender, DM, smoking status, SBP, and TC;(4)Age, gender, DM, smoking status, SBP, TC, and HDL;(5)Age, gender, DM, smoking status, SBP, HDL, and LDL;(6)Age, gender, DM, smoking status, SBP, and LDL.The TCVR score was developed to estimate the ten-year risk of a major cardiovascular event in the Thai population based on the data of the patient which included age, gender, diabetic status (DM), SBP, WC, height, TC, HDL, and LDL. The TCVR can be calculated using six models. The first two models do not require any laboratory data and the last four models require some combination of lipid profiles. Six different models were generated to encourage the use of the score based on the availability of patient data. In some models, no laboratory results were required. This is because some investigations are not covered by the Thai Universal Healthcare Coverage. Thus, it is possible that some patients do not have all lipid profile values ,30. The p-value of <0.05 was considered statistically significant.Descriptive analysis was used to describe participants’ characteristics. The association between patients’ demographic and MCI status was analyzed using Chi’s square and T-test. We created six prediction models using the variables presented in the six TCVR score models. A multivariable analysis was used. Each prediction model was evaluated using the area under the receiver operator characteristic curve (AuROC) to explore their clinical usefulness. The Hosmer–Lemeshow goodness of fit test was used for the determination of model calibration. For sensitivity analysis, every TCVR model was then applied to a subgroup of the population stratified by age (<65 or ≥65) and gender, as evidence has suggested that discriminative performance for cognitive performance may be modified by age and gender . The subThe missing data are presented in p = 0.017 and 19.54 ± 3.44 vs. 26.21 ± 1.21, p < 0.001, respectively). There were no significant differences in other characteristics and laboratory results across the categories of MCI status. There were only eight smokers in the sample, all with MCI; thus, this variable could not be used for model development.p = 0.286; Model 2 X2 = 13.55, df = 8, p = 0.094; Model 3 X2 = 5.84, df = 8, p = 0.665; Model 4 X2 = 4.54, df = 8, p = 0.805; Model 5 X2 = 2.17, df = 8, p = 0.975; and Model 6 X2 = 6.73, df = 8, p = 0.566.Six models using different TCVR variables were developed. The adjusted odds ratio of each model is presented in In the sensitivity analysis stratified by age and gender, the discriminative performance of models that required only anthropometric data is shown in In our sample, the prevalence of MCI among patients with metabolic risks was as high as 82.7 percent. The discriminative performance (AuROC) of six different TCVR models utilizing commonly measured variables for CV risk assessment varied from 0.58 to 0.61. In the subgroup analysis, the anthropometric-based models demonstrated better discriminative performance in the older age group (≥65), whereas laboratory-based models provided higher discriminative performance for the younger age group (<65). However, the discriminative ability of the six models is still relatively poor and further model development and validation studies are needed to help better assess the risk of developing MCI for people with metabolic risks.Detecting early stages of cognitive impairment such as MCI or early dementia might help physicians to provide better care for people with metabolic risks. The high prevalence of MCI in our sample is similar to the results of recent studies showing that the prevalence of MCI in individuals with metabolic risk factors was approximately between 65 and 75 percent ,34. ThisThe relatively poor discriminative property may be due to the limitation in identifying the exact causes of MCI. The underlying mechanism could be more than vascular etiology which is commonly associated with metabolic disease . Non-vas2 or over for both genders, the WC cut-off value of 90 cm or over for men, and 82 cm or over for women were indicators for screening older adults who were at risk for MCI [Interestingly, the algorithm without laboratory parameters was able to better predict the probability of MCI in older age individuals, especially in men. An independent risk factor for cognitive impairment in elderly people is anthropometric measurement. Obesity indicators, including body mass index and WC, are linked to an increased risk of cognitive decline. One study showed that the body mass index cut-off value of 26 kg/m for MCI . Further for MCI .The model that included laboratory results, TC and HDL, on the other hand, appeared to be more beneficial in younger individuals. Several cardiovascular risk factors, including smoking, diabetic status, hypertension, and serum cholesterol in middle age, have previously been linked to late-life dementia ,51. In aThis study demonstrated that there is the potential to develop an MCI risk assessment tool among patients with metabolic syndrome using the measurements obtained from routine cardiovascular risk assessments. Our results are in agreement with previous studies. The cardiovascular risk profile was linked to a decline in cognitive function and could be seen at as early as 35 years of age . AnotherPatients with metabolic risk factors are at a higher risk for both major cardiovascular events and cognitive impairment. Early diagnosis of MCI could lead to the development of a treatment for prevention and delay the progression of the disease. This study suggests that the values and results obtained from routine cardiovascular risk assessment could potentially be used to develop a screening tool for MCI. However, as the discriminative ability of the six models is still relatively poor and may only be acceptable by using different models for different subgroups, further model development, and validation studies are needed.