VEGFA) and CXC chemokines have been shown to play vital roles in angiogenesis. Exploring the expression level, gene regulatory network, prognostic value, and target prediction of the CXC chemokine-VEGFA network in colon adenocarcinoma (COAD) is crucial from the perspective of tumor angiogenesis.  Tumor angiogenesis plays a vital role in tumorigenesis, proliferation, and metastasis. Recently, vascular endothelial growth factor A , TRRUST (version 2), LinkedOmics, and Metascape). CXCL1/2/3/5/6/8/11/16/17 and VEGFA were markedly overexpressed, while CXCL12/13/14 were underexpressed in patients with COAD. Moreover, genetic alterations in the CXC chemokine-VEGFA network found at varying rates in patients with COAD were as follows: CXCL1/2/17 (2.1%), CXCL3/16 (2.6%), CXCL5/14 (2.4%), CXCL6 (3%), CXCL8 (0.8%), CXCL11/13 (1.9%), CXCL12 (0.6%), and VEGFA (1.3%). Promoter methylation of CXCL1/2/3/11/13/17 was considerably lower in patients with COAD, whereas methylation of CXCL5/6/12/14 and VEGFA was considerably higher. Furthermore, CXCL9/10/11 and VEGFA expression was notably correlated with the pathological stages of COAD. In addition, patients with COAD with high CXCL8/11/14 or low VEGFA expression levels survived longer than patients with dissimilar expression levels. CXC chemokines and VEGFA form a complex regulatory network through coexpression, colocalization, and genetic interactions. Moreover, many transcription factor targets of the CXC chemokine-VEGFA network in patients with COAD were identified: RELA, NFKB1, ZFP36, XBP1, HDAC2, SP1, ATF4, EP300, BRCA1, ESR1, HIF1A, EGR1, STAT3, and JUN. We further identified the top three miRNAs involved in regulating each CXC chemokine within the network: miR-518C, miR-369-3P, and miR-448 regulated CXCL1; miR-518C, miR-218, and miR-493 regulated CXCL2; miR-448, miR-369-3P, and miR-221 regulated CXCL3; miR-423 regulated CXCL13; miR-378, miR-381, and miR-210 regulated CXCL14; miR-369-3P, miR-382, and miR-208 regulated CXCL17; miR-486 and miR-199A regulated VEGFA. Furthermore, the CXC chemokine-VEGFA network in patients with COAD was notably associated with immune infiltration.  Our results showed that  This study revealed that the CXC chemokine-VEGFA network might act as a prognostic biomarker for patients with COAD. Moreover, our study provides new therapeutic targets for COAD, serving as a reference for further research in the future. Colon cancer is a common malignant tumor of the digestive tract. The incidence and mortality of colon adenocarcinoma (COAD) are the third highest of all cancer types . Since tVEGFA) is a vital factor that plays an essential role in tumor angiogenesis and development [VEGFA inhibitor, has been used to treat advanced renal cell carcinoma. However, the side effects of sunitinib can be quite severe and include kidney and cardiovascular damage [VEGFA are heavily regulated during tumor angiogenesis. CXCL12 can promote a malignant phenotype by promoting the clonal growth of colorectal cancer cells and regulating the expression of VEGF and ICAM-1 [Chemokines are a family of small heparin-binding proteins 8–10 kDa in size. Four subgroups exist within the chemokine family . The CXC subgroup has been shown to play a crucial role in angiogenesis in physiological and pathological settings . Recentlr damage . CXC cheVEGFA expression and the development and prognosis of COAD, as well as to provide new insights into targeted therapies for patients with COAD.Multiple online databases were used to explore the expression level, gene regulation network, prognostic value, and regulation targets of the CXC chemokine-VEGFA network in patients with COAD from an angiogenic perspective in this study. In addition, we aimed to identify the relationship between CXC chemokine and http://ualcan.path.uab.edu/analysis.html) is a free online database that provides analysis based on The Cancer Genome Atlas (TCGA) and MET500 cohort data [VEGFA, (2) dataset: COAD, and (3) threshold setting conditions: P value cutoff = 0.05. A Student's t-test was used for the comparative analysis [UALCAN , an open-access resource, provides analyses of specific human genes and proteins [VEGFA, (2) section: tissue and pathology, (3) tissue: colon and COAD, and (4) picture of tissue types: normal colon tissue and COAD. Data were obtained on February 14, 2022.The Human Protein Atlas (proteins . Screenihttp://gepia.cancer-pku.cn/index.html) is an analysis tool that delivers RNA sequencing expression data from 9,736 cancerous and 8,587 noncancerous samples [VEGFA), dataset (COAD), and threshold conditions  were set as screening criteria. The expression of CXC chemokines and VEGFA, as well as the pathological stage of COAD, was analyzed using a Student's t-test. The prognosis of patients with COAD was analyzed using the Kaplan–Meier curve [GEPIA  is a free online database for visualizing, studying, and analyzing cancer genomic data [z-score threshold of ±2.0 was used to calculate mRNA expression z-scores for all samples (log RNA Seq V2 RSEM). CXC chemokines and VEGFA were the chosen genes [cBioPortal (mic data . The anaen genes –16. Datahttps://string-db.org/cgi/input.pl) is a free online database that helps researchers analyze all publicly available sources of protein–protein interaction (PPI) data [Homo sapiens [STRING (PI) data . We crea sapiens , 15. Dathttp://www.genemania.org) is a free online database that creates PPI networks and analyzes gene function [VEGFA [GeneMANIA (function . The intn [VEGFA –16. Datahttps://metascape.org) is a free online gene function analysis tool that assists users in using current common bioinformatics analysis approaches to batch gene and protein analysis to predict function [Metascape (function . We condfunction –16. Datahttps://www.grnpedia.org/trrust/) is a free online database for human transcriptional regulatory networks [VEGFA) were chosen in this study [TRRUST (networks . We sougis study –16. Datahttp://www.linkedomics.org/) is a free database that provides methods for analyzing and comparing cancer multiomics data [VEGFA), and target dataset (RNA-seq) were chosen in this study [LinkedOmics (ics data . The “Liis study –16. Datahttps://cistrome.shinyapps.io/timer/) is a free online platform for systematically analyzing tumor-infiltrating immune cells [TIMER (ne cells . The “Gene cells –16. DataCXCL1/2/3/5/6/8/11/16/17 and VEGFA were remarkably upregulated in (1) sex , (2) pathological stage (stage 1–4), and (3) sample type (COAD) (P < 0.05)  sex , (2) pathological stage (stage 1–4), and (3) sample type (COAD) (P < 0.05)  sex , (2) pathological stage (stage 2), and (3) sample type (COAD) (P < 0.05)  (CXCL8/11/14 expression were higher (P ≤ 0.05)   . Subsequ Figures  or when  < 0.05) .VEGFA was altered by 1.3% in COAD patients , CXCL3/16 (2.6%), CXCL5/14 (2.4%), CXCL6 (3%), CXCL8 (0.8%), CXCL11/13 (1.9%), and CXCL12 (0.6%), were found  signaling pathway . In addition, CXCL8 and VEGFA were found to be regulated by ZFP36 ring finger protein (ZFP36), X-box-binding protein 1 (XBP1), histone deacetylase 2 (HDAC2), activating transcription factor 4 (ATF4), E1A-binding protein p300 (EP300), early growth response 1 (EGR1), signal transducer and activator of transcription 3 (STAT3), and Jun proto-oncogene (JUN) (P < 0.01). Furthermore, CXCL1/5/14 and VEGFA were found to be regulated by Sp1 transcription factor (SP1) (P < 0.001). Breast cancer 1 (BRCA1) was the key transcription factor involved with CXCL1 and VEGFA in COAD patients (P < 0.01). Finally, estrogen receptor 1 (ESR1) and hypoxia-inducible factor 1 alpha subunit (HIF1A) regulated the functions of CXCL12 and VEGFA (P < 0.01).Potential transcription factors involved with the CXC chemokine-VEGFA network in COAD patients were identified . v-rel rCXCL1 were miR-518C, miR-369-3P, and miR-44. In addition, miR-518C, miR-218, and miR-493 were identified as potential miRNA targets that regulate CXCL2. Furthermore, we observed that CXCL3 was regulated by miR-448, miR-369-3P, and miR-221. miRNA target of CXCL13 is miR-423. Moreover, miR-378, miR-381, and miR-210 were identified as potential miRNA targets that regulate CXCL14. CXCL17 is regulated by miR-369-3P, miR-382, and miR-208. Furthermore, our results showed that miR-486 and miR-199A are potential miRNA targets that regulate VEGFA.The top three miRNA targets of the CXC chemokine-VEGFA network were obtained . The miRCXCL1/2/3/5/6/8/11/12/13/14/16/17 and VEGFA   = 0.8921, P = 4.226e–132)    , CXCL2 (304e–90) , and ZC3882e–47) .CXCL2 expression, respectively                                      , CXCL1 (304e–90) , and ZC3735e–46) . Furtherectively . Among t Figures . Express26e–132) , CXCL2 (01e–119) , and ZC3446e–51) . Our resectively . Among t884e–68) , IL8 (PC632e–63) , and MMP269e–62) . Our resectively . Among t Figures . CXCL6 e689e–59) , MMP3 (P921e–55) , and IL8935e–53) . In addiectively . Among t Figures . CXCL8 e939e–76) , IL1B (P.25e–73) , and OSM368e–72) . Furtherectively . Among t Figures . CXCL11 99e–101) , UBD (PC935e–62) , and IDO137e–60) . Moreoveectively . Among t Figures . CXCL12 835e–87) , SLIT3 (915e–86) , and SHE928e–84) . Our resectively . Among t Figures . CXCL13 598e–89) , SH2D1A 229e–80) , and SIR508e–80) . In addiectively . Among t Figures . CXCL14 .24e–58) , TNFSF11643e–41) , and COL338e–41) . Furtherectively . Among t Figures . CXCL16 175e–83) , FLII (P248e–40) , and NDE486e–39) . We alsoectively . Among t Figures . CXCL17 148e–21) , GPR110 333e–19) , and SEM753e–16) . Finallyectively . Among t Figures . VEGFA e639e–35) , CCNL1 (411e–30) , and CRE606e–27) .CXCL1 expression in COAD patients was positively associated with CD8+ T cell infiltration, neutrophils, and dendritic cells (P < 0.05) (CXCL1 expression (P < 0.01) (CXCL2 and CXCL3 (P < 0.001)     (CXCL17 expression (P < 0.001) (VEGFA expression (P < 0.01) ( < 0.05) . However < 0.01) . In addi Figures . However Figures . Further Figures . B cells Figures . The exp < 0.05) . B cells< 0.001) . CD4+ T  < 0.01) .VEGFA and CXC chemokines as important participants in angiogenesis, particularly tumor angiogenesis [VEGFA have been studied in a range of tumor types; however, findings are contradictory with regard to colonic adenocarcinomas [Tumor angiogenesis plays a vital role in tumorigenesis, proliferation, and metastasis. In recent years, studies have identified ogenesis , 26–28. rcinomas , 30. ThiCXCL1/2/3/5/6/8/11/16/17 and VEGFA was upregulated in patients with COAD compared with that in individuals without COAD. Patients with COAD also showed downregulated CXCL12/13/14 expression. The results were similar to those reported in a previous study in patients with COAD [CXCL5/6/12/14 and VEGFA were higher in patients with COAD than those in healthy individuals. Conversely, the promoter methylation levels of CXCL1/2/3/11/13/17 were lower in patients with COAD. Thus, we hypothesized that genetic methylation and alteration within the CXC chemokine-VEGFA network might be the leading cause of abnormal gene expression levels in patients with COAD.In this study, we also examined the potential correlation between pathological stage and differential expression of COAD. The expression of ith COAD  and contith COAD . This maCXCL9/10/11 and VEGFA expression and the pathological stage of COAD. Furthermore, the survival of patients with COAD was higher with low VEGFA or high CXCL8/11/14 expression levels. Therefore, the expression levels of CXCL8/11/14 and VEGFA may be potential prognostic indicators for COAD. CXCL8/11/14 and VEGFA promote tumor angiogenesis in different ways [We also observed a notable correlation between the ent ways –33. ThusVEGFA may promote cancer progression, and this could be through a potential interaction network. Genes in the network were mainly involved in cytokine receptor binding, chemokine and cytokine activity, leukocyte chemotaxis, and migration, all of which are closely related to angiogenesis. For instance, IL-8 (CXCL8) promotes tumor angiogenesis by binding to CXCR1 and CXCR2 receptors [The potential functions and interactions of the CXC chemokine-VEGFA network were further explored in this study. They were found to be complex and tightly connected. CXC chemokines and eceptors . In addieceptors . CollectκB signaling pathway were highly involved in the CXC chemokine-VEGFA network in COAD patients, all of which are highly related to tumor angiogenesis [Furthermore, the functions of the CXC chemokine-VEGFA network in patients with COAD were mainly related to chemokine activity, cytokine activity, and growth factor activity, as demonstrated by GO enrichment analysis, all of which are closely related to tumor angiogenesis. More studies are needed to confirm the mechanism by which this happens. In this study, we further found through KEGG pathway analysis that the cytokine–cytokine receptor interaction signaling pathway, IL-17 signaling pathway, and NF-ogenesis , 37. TheVEGFA. Studies have shown that RELA, NFKB1, HDAC2, SP1, ATF4, EP300, BRCA1, ESR1, HIF1A, EGR1, STAT3, and JUN regulate tumor angiogenesis, thus affecting tumor growth and prognosis [Mutated or altered transcription factors represent a unique class of drug targets that mediate aberrant gene expression, and the development of corresponding targeting drugs may impact future cancer treatments. Thus, the targets and regulators of the CXC chemokine-VEGFA network in COAD patients were further analyzed. The transcription factor targets of the CXC chemokine-VEGFA network in patients with COAD were identified. RELA, NFKB1, ZFP36, XBP1, HDAC2, SP1, ATF4, EP300, BRCA1, ESR1, HIF1A, EGR1, STAT3, and JUN were deemed crucial regulatory factors. Our results showed that these factors have potential functions in regulating tumor angiogenesis by targeting rognosis , 38–48. rognosis –52. In sVEGFA. Some of the genes with the highest correlation  were positively associated with tumor angiogenesis [VEGFA-related regulatory targets may serve as a viable therapeutic oncology approach.The correlation between CXC chemokine-VEGFA network expression and differentially expressed genes in COAD patients was explored in this study. We found that in patients with COAD, approximately 20,000 genes were negatively or positively correlated with CXC chemokine-VEGFA network expression. From these, we screened for genes with the highest correlation with CXC chemokines and ogenesis , 54. RegIn this study, we determined the expression levels and gene regulatory network of the CXC chemokine-VEGFA network, which plays a vital role in angiogenesis in COAD. We also identified new prognostic biomarkers and therapeutic targets. These findings provide insight into the study and treatment of COAD.


Ceftazidime-avibactam (caz-avi), a novel β-lactam/β-lactamase inhibitor, is commonly utilized for carbapenem-resistant gram-negative infections (CR-GNI). However, the benefits vs risks of combining caz-avi with other agents are unclear.In this retrospective cohort study, inpatients with CR-GNI treated with caz-avi were identified at 9 U.S. hospitals. The impact of caz-avi monotherapy (MT) or combination therapy  was studied using logistic regression, controlling for baseline patient and hospital factors. The primary outcome was in-hospital mortality or discharge to hospice (death), and secondary outcomes were length of stay (LOS), resolution of infectious signs and symptoms , 90-day recurrent infection and future caz-avi–resistant organism. An adjusted odds ratio (aOR) with 95% confidence interval (CI) was used to assess the primary and secondary outcomes.Klebsiella spp. (44.6%) followed by Pseudomonas aeruginosa (27.7%) (table 2). Concomitant gram-negative agents are shown in table 3. Overall, 92 (28.1%) patients died and CT (vs MT) displayed similar adjusted mortality risk  and LOS . CT (vs MT) was associated with greater odds of clinical response (aOR: 2.25 [95%CI:1.15-4.41]). Among survivors, similar rates of 90-day recurrent infection (50/154 (32.5%) were observed in CT vs 18/82 (22.0%) in MT group (p=0.09) and 5 (2.19%) patients had future infection with a caz-avi–resistant pathogen (3 in CT and 2 in MT group).328/499 (65.7%) patients received caz-avi as targeted therapy for a CR-GNI. Overall patients treated with MT and CT were similar at baseline and had comparable baseline demographics although patients treated with CT were more likely to be in the ICU and receive a concomitant empiric in vitro-concordant antibiotic (table 1). The most common organism was Compared to patients with CR-GNI treated with caz-avi alone, those who received CT including caz-avy had similar survival and LOS but higher clinical response. The role of CT in the era of novel antibiotics warrants additional study.Helen W. Boucher, MD, American Society of Microbiology: Honoraria|Elsevier: Honoraria|Sanford Guide: Honoraria.


Aspergillus fumigatus (AFM) is mainly associated with mutations in cyp51A and its promoter region or its homologue cyp51B. We evaluated the in vitro activity of isavuconazole (ISC), itraconazole (ITC), posaconazole (PSC), and voriconazole (VRC) against 660 AFM collected during 2017–2020.Azole resistance in cyp51 genes using whole genome sequencing.Isolates from Europe (EU), North America (NA), Latin America (LA), and Asia-Pacific (APAC) were identified by MALDI-TOF and/or sequencing and tested by CLSI broth microdilution. CLSI epidemiological cut-off values (ECV) were applied. A PSC ECV of 0.5 mg/L was used. Non-wildtype (NWT) isolates to azoles were screened for alterations in the cyp51 genes. Of these AFM, 29/32 (90.1%) were NWT to ITC, 25/32 (78.1%) NWT to ISC, 17/32 (53.1%) NWT to VRC, and 11/32 (34.4%) NWT to PSC. The most frequent alteration was CYP51A TR34/L98H, carried by 14 EU isolates, all NWT to ISC and ITC. Four isolates from NA carried the alteration I242V in CYP51A . One NA isolate carried the CYP51A alteration G448S  and one carried A9T (NWT to ISC). A single isolate from APAC carried a CYP51A G138C alteration and was NWT to all 4 triazoles. Multiple alterations in CYP51A were detected in 5 isolates: 4/5 NWT to ISC or ITC, 1/5 NWT to VRC, all WT to PSC. Alterations in CYP51B were noted in 7 isolates; 6/7 carried Q42L, 3 from NA , 2 from APAC , and 1 from EU . Among 34 NWT isolates without cyp51 alterations, 32.4% were WT to ISC, 47.1% WT to ITC, 85.3% WT to VRC, and 82.4% WT to PSC.Azoles had similar activities against 660 AFM isolates. Overall, AFM displayed WT MIC values to ISC (92.7%), ITC (92.9%), PSC (97.3%), and VRC (96.7%). Only 66 isolates (10.0%) were NWT to 1 or more of the azoles, and 32 harbored one or more alterations in the cyp51 alterations were detected in 32/66 NWT isolates. Only EU isolates harboured the environmental alteration TR34/L98H that was associated with a NWT phenotype to ISC and ITC. Alterations in AFM cyp51 can have variable effects on the in vitro activity of the azoles that are best delineated by testing all triazoles.The majority of AFM were WT to azoles. Ten different Michael A. Pfaller, MD, Pfizer: Grant/Research Support Cecilia G. Carvalhaes, MD, PhD, AbbVie: Grant/Research Support|Cidara: Grant/Research Support|Melinta: Grant/Research Support|Pfizer: Grant/Research Support Lalitagauri M. Deshpande, PhD, Melinta: Grant/Research Support|Pfizer: Grant/Research Support Paul Rhomberg, BS, MT(ASCP), Cidara: Grant/Research Support|Pfizer: Grant/Research Support Paul Rhomberg, BS, MT(ASCP), Cidara: Grant/Research Support|Pfizer: Grant/Research Support.


Lenvatinib has shown promising efficacy in targeted therapies that have been tested to treat anaplastic thyroid carcinoma (ATC) in both preclinical and clinical studies. The aim of this study was to evaluate the efficacy and safety of lenvatinib in the treatment of patients with ATC.PubMed, the Cochrane Library, Embase, and ClinicalTrials.gov were searched for potential eligible studies from inception to February 1, 2022. The outcomes included partial response (PR), stable disease (SD), disease control rate (DCR), median progression-free survival (mPFS), and median overall survival (mOS). Effect sizes for all pooled results were presented with 95% CIs with upper and lower limit.Ten studies met the inclusion criteria. The aggregated results showed that the pooled PR, SD, and DCR were 15.0%, 42.0%, and 63.0%, respectively. The pooled mPFS and mOS were 3.16 (2.18–5.60) months and 3.16 (2.17–5.64) months, respectively. Furthermore, PFS rate at 3 months (PFSR-3m), PFSR-6m, PFSR-9m, PFSR-12m, and PFSR-15m were 52.0%, 22.5%, 13.9%, 8.4%, and 2.5%, respectively. Meanwhile, the 3-month OS rate (OSR-3m), OSR-6m, OSR-9m, OSR-12m, and OSR-15m were 64.0%, 39.3%, 29.7%, 18.9%, and 14.2%, respectively. The most common adverse events (AEs) of lenvatinib were hypertension (56.6%), proteinuria (32.6%), and fatigue (32%).This meta-analysis showed that lenvatinib has meaningful antitumor activity, but limited clinical efficacy in ATC.https://www.crd.york.ac.uk/PROSPERO/], identifier [CRD42022308624].PROSPERO [ Anaplastic thyroid carcinoma (ATC), a malignancy derived from undifferentiated thyroid follicular cells , accountLenvatinib is a multi-target antiangiogenetic broad-spectrum tyrosine kinase inhibitor (TKI) that can inhibit various signal receptors  –12. In aWe have registered our protocol on PROSPERO (registration number: CRD42022308624). This meta-analysis followed the Preferred Reporting Items for Systematic Review and Meta-Analysis (PRISMA) statement . The PRIPubMed, the Cochrane Library, Embase, and ClinicalTrials.gov were searched for potential eligible studies. The search was performed from inception to February 1, 2022. The search keywords were “thyroid carcinoma, anaplastic” and “lenvatinib” and the search strategy in PubMed was as follows: Thyroid Carcinoma, Anaplastic [Mesh] OR Anaplastic Thyroid Carcinoma [Title/Abstract] OR Anaplastic Thyroid Carcinomas [Title/Abstract] OR Carcinoma, Anaplastic Thyroid [Title/Abstract] OR Carcinomas, Anaplastic Thyroid [Title/Abstract] OR Thyroid Carcinomas, Anaplastic [Title/Abstract] OR Thyroid Cancer, Anaplastic [Title/Abstract] OR Anaplastic Thyroid Cancer [Title/Abstract] OR Anaplastic Thyroid Cancers [Title/Abstract] OR Cancer, Anaplastic Thyroid [Title/Abstract] OR Cancers, Anaplastic Thyroid [Title/Abstract] OR Thyroid Cancers, Anaplastic [Title/Abstract] AND Lenvatinib [Mesh] OR 4-(3-chloro-4-((cyclopropylaminocarbonyl)amino)phenoxy)-7-methoxy-6-quinolinecarboxamide [Title/Abstract] OR N-(4-((6-carbamoyl-7-methoxyquinolin-4-yl)oxy)-2-chlorophenyl)-N’-cyclopropylurea [Title/Abstract] OR 4-(3-chloro-4-(N’-cyclopropylureido)phenoxy)-7-methoxyquinoline-6-carboxamide [Title/Abstract] OR lenvatinib mesylate [Title/Abstract])) OR (E7080 mesylate [Title/Abstract] OR monomethanesulfonate [Title/Abstract] OR lenvatinib mesylate [Title/Abstract] OR lenvatinib methanesulfonate [Title/Abstract] OR Lenvima [Title/Abstract] OR E-7080 mesylate [Title/Abstract] OR E 7080 [Title/Abstract] OR 4-(3-chloro-4-(((cyclopropylamino)carbonyl)amino)phenoxy)-7-hydroxy-6-quinolinecarboxamide [Title/Abstract] OR E-7080 [Title/Abstract] OR ER-203492-00 [Title/Abstract] OR E7080 [Title/Abstract] OR lenvatinib metabolite M2 [Title/Abstract]. No language, region, ethnicity, age, or payment restrictions were imposed during the search process.Inclusion criteria were as follows : studiesMethodological index for non-randomized studies (MINORS) evaluates single-arm studies . JBI CriTwo investigators independently made study selection. If there were any differences between them, the third author would discuss with them together. Information on the following characteristics of included studies was recorded: authors, study type, sample size, age, criteria for tumor response , adverse events (AEs), and reported endpoints.I2 statistic. When I2 ≤ 50%, use the fixed-effects model; otherwise, use the random-effects model. For pooled results with high heterogeneity, the sensitivity analysis was performed by excluding each study individually. Begg’s test, Egger’s test, and the trim-and-fill method were used to assess publication bias. p < 0.05 was considered statistically significant.Analysis of pooled PR, SD, and DCR, and of the pooled K-M curves of ATC patients treated with lenvatinib was performed using R version 3.6.3. Effect sizes for all pooled results were presented with 95% CIs with upper and lower limit. Heterogeneity between studies was examined using the Cochrane Q chi-square test and We initially identified 349 studies. Finally, our study included 10 studies, namely, 2 prospective studies , 18 and Two single-arm studies , 18 scorI2 = 59.0%, p < 0.01), and the pooled PR in subgroups was different , while the other subgroups showed a pooled PR of 11% .We extracted efficacy measures from each study which included in this meta-analysis , while the subgroup of the retrospective study showed a pooled SD of 36% , and the subgroup of the prospective study resulted in a pooled SD of 59%   and 59% , respectively. The total pooled DCR was 63%   on the estimated pooled PR and DCR , Tianjin Municipal Science and Technology Project  and Beijing-Tianjin-Hebei Basic Research Cooperation Project (20JCZXJC00120), The Science & Technology Development Fund of Tianjin Education Commission for Higher Education (2021ZD033), Tianjin Medical Key Discipline  Construction Project.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.


Correction:J ExpClin Cancer Res41, 299 (2022)https://doi.org/10.1186/s13046-022-02501-3Following publication of the original article , author This correction does not change the result, interpretation, and conclusions of the study. The original article has been corrected.Additional file 1: Supplementary Figure S5. Blank and plasma membrane impregnated MB morphology by SEM (25.13 K magnification). Scale bar = 1 μm.


Nucleic acid reagents, including plasmid-encoded genes and small interfering RNA (siRNA), are promising tools for validating gene function and for the development of therapeutic agents. Native β-cyclodextrins (BCDs) have limited efficiency in gene delivery due to their instable complexes with nucleic acid. We hypothesized that cationic BCD nanoparticles could be an efficient carrier for both DNA and siRNA. Tetraethylenepentamine-coated β-cyclodextrin (TEPA-BCD) nanoparticles were synthesized, characterized, and evaluated for targeted cell delivery of plasmid DNA and siRNA. The cationic TEPA coating provided ideal zeta potential and effective nucleic acid binding ability. When transfecting plasmid encoding green fluorescent protein (GFP) by TEPA-BCD, excellent GFP expression could be achieved in multiple cell lines. In addition, siRNA transfected by TEPA-BCD suppressed target GFP gene expression. We showed that TEPA-BCD internalization was mediated by energy-dependent endocytosis via both clathrin-dependent and caveolin-dependent endocytic pathways. TEPA-BCD nanoparticles provide an effective means of nucleic acid delivery and can act as potential carriers in future pharmaceutical application. Gene expression could be modulated through the use of exogenous nucleic acids. The power of nucleic-acid-based drugs, including DNA molecules and small interfering RNAs (siRNAs), lies in their abilities to specifically enhance or silence genes of interest. The siRNA-based drugs were recently approved by US Federal Drug Administration , markingNonviral gene delivery vectors have been shown to have several advantages, such as ease of synthesis, mass production, versatile modification, and low immunogenicity . There aCationic polysaccharides can bind with siRNAs to form complexes, thereby protecting siRNA from degradation and neutralizing their negative charges . NanoparIn this study, we developed an easy and efficient strategy to generate TEPA-BCD. The 6-hydroxyl groups of BCD were activated with tosyl chloride. After nucleophilic displacement by ethylenediamine (EDA), TEPA-BCD nanoparticles were synthesized by the crosslinking reaction of EDA-BCD and TEPA via glutaraldehyde (GA). The synthesis of TEPA-BCD nanoparticles is demonstrated in −1 , 1036 cm−1 (C–O vibration). Another two intense peaks around 1036 and 2900 cm−1 from C-O and C–H stretching were also found in all BCD derivatives indicating the main BCD structure was maintained after the amine introduction .,45.44,45CPZ blocked clathrin-coated pit formation , and conDevelopment of siRNA delivery systems depends not only on their transfection efficiencies, but also on their gene-silencing effect. Although both Gen and MCD treatments showed no reduction in plasmid DNA transfection efficiency C, the geIn the intravenous delivery, the nanoparticles have to escape the various barriers such as kidney filtration, phagocytosis, and hydrolysis degradation. Sequentially, they should transport across vascular vessels, diffuse through the extracellular matrix, and reach the target cells . The enhIn summary, we developed a cationic BCD-based nanoparticle delivery system and applied it in the fibroblast 3T3 cell line and the epithelial ARPE cell line. Our results demonstrated the multifunctionality critical for internalization and activity of both gene-coded plasmid DNA and siRNA. When the TEPA-BCD/plasmid ratio was 1.71:1, the nanoparticles had a transfection efficiency of 97%, and cells maintained viability around 83%. When the TEPA-BCD/siRNA ratio was 96:1, it knocked down the fluorescent expression by 57%, yet the cell viability remained around 82%. TEPA-modified BCD demonstrated a nanometer size, positive surface charge, minimal cytotoxicity, great electrostatic interaction with nucleic acid, and efficient delivery. In both ARPE cells and 3T3 cells, TEPA-BCD entered cells through clathrin- and caveolae-mediated endocytosis. TEPA-BCD/plasmid DNA mediated high gene expression and TEPA-BCD/siRNA displayed efficient gene silencing. The TEPA-BCD provides a promising platform for delivery of therapeutic nucleic acids in vitro and warrants further development of its potential for in vivo delivery.BCD was a generous gift from Feng-Yuan Biotech . Acetone, methanol, and ethanol were purchased from Echo Chemicals . Green fluorescent protein (GFP)-expressing plasmid was obtained from Takara Bio . Fetal bovine serum (FBS) was purchased from Biological Industries . The siRNA against GFP gene  was purchased from MDBio . DNA and siRNA transfection controls (PolyJet and GenMute) are purchased from SignaGen . TEPA, p-tolylsulfonyl chloride, sodium chloride, and other chemicals were purchased from Sigma-Aldrich . All reagents were used without further purification.1H NMR  of BCD: δ = 5.69 , 4.84 , 4.44 , 3.62 , 3.33  ppm. 13C NMR  of BCD: δ = 102.04 (C-1), 82.01 (C-4), 73.51–72.51 , 69.63 , 60.37 (C-6) ppm. 1H NMR  of Tosyl-BCD: δ = 7.12 , 7.45 , 5.70 , 4.80 , 4.50 , 4.32 , 4.21 , 3.48 , 2.43  ppm. 13C NMR  of Tosyl-BCD: δ = 128.48 , 102.44 (C-1), 82.02 (C-4), 73.55–72.50 , 60.41 (C-6), 31.16 (Tosyl-CH3) ppm. 1H NMR  of EDA-BCD: δ = 5.70 , 4.84 , 4.44 , 3.48 , 2.1  ppm. 13C NMR  of EDA-BCD: δ = 101.82 (C-1), 81.09 (C-4), 73.04–71.78 , 60.25 (C-6), 42.04 (BCD-C-C-N) ppm. 1H NMR  of TEPA-BCD: 5.06 , 4.77 , 4.47 , 3.74 , 3.19 , 2.84 , 2.22  ppm. 13C NMR  of TEPA-BCD: δ = 102.10 (C-1), 81.24 (C-4), 73.20–72.08 , 60.15 (C-6), 45.22 (C-NH), 30.24 (cyclic ether C-C-O) ppm.Mono-6-deoxy-6-p-tolylsulfonyl (tosyl)-BCD and EDA-BCD were synthesized as described previously ,50. BrieE. coli cells. The host E. coli was grown in LB broth with kanamycin (50 µg/mL) and the plasmid DNA was isolated using Mini Purification Kit . The concentration and quality of nucleic acids were determined using a NanoDrop spectrophotometer  and the Beer–Lambert law with an extinction coefficient of 50 μg/mL−1 cm−1 at 260 nm. Isolated plasmid DNA had OD 260/280 ratios of 1.80–1.95, indicating the quality was suitable for our application. Polyplexes were formulated by mixing 0.21–1.71 μg (10–80 μL) TEPA-BCD with plasmid DNA (1 μg) or siRNA  in 100 μL DMEM medium.We transformed plasmid GFP-C3 into DH-5α To visualize morphology of BCD derivatives, diluted samples were adsorbed onto a 100 mesh copper grid and dried overnight, followed by imaging on a transmission electron microscope . The magnification power of TEM is 50,000. The chemical groups of modified BCDs were investigated using Fourier transform infrared spectroscopy  and an NMR spectrometer (Bruker AV III HD). The size and zeta potentials of the TEPA-BCD complexes were measured using ZetaSizer with 633 nm He-Ne Laser  and the folded capillary cell .ARPE cells  and 3T3 cells (a mouse embryonic fibroblast cell line) from the cell bank of BCRC  were used for GFP plasmid delivery experiments. DNA transfection was performed as previously reported . APRE-GF4 cells/well. When reaching 90% confluence, the cells were treated with polyplex and endocytic inhibitor for 3 h. Polyplexes were formulated by mixing 1.28 μg TEPA-BCD with plasmid DNA (1 μg) or siRNA (13.3 ng) in 100 μL DMEM medium. Subsequently, the transfection medium was replaced with fresh DMEM supplemented with 10% FBS and cells were incubated for another 24 h. Imaging was conducted on an automated image analyzer for analysis of fluorescence intensity and cell viability.The cellular uptake mechanisms of polyplex were examined using different endocytic inhibitors: chlorpromazine (30 µM), genistein (200 µM), sodium azide (10 mg/mL), methyl-BCD (10 mM), and monensin (3 µM). The cells were seeded in 48-well plates at 5 × 10t-test by Analysis Toolpak in Microsoft Excel. Statistical significance is represented as * p < 0.05.All cellular data were performed by two individual experiments. The Zetasizer results were the average from three individual experiments. The mean and standard deviation were calculated using Microsoft Excel. Statistical comparisons are performed using PROC ANOVA followed by the Tukey post hoc test using SAS 9.4 software  and unpaired two-tailed Student


In the United Kingdom(1) and internationally(2), help-seeking for domestic violence (DV) and domestic homicides have increased(3) during COVID-19 lockdown periods. Suspension and remote delivery of face-to-face clinical services, continuing healthcare and other support services limits opportunities for DV detection and disclosure.This presentation will summarise changes in DV incidence and help-seeking during COVID-19, their impacts on health and wellbeing, and present guidance for clinicians assessing and supporting survivors.World Health Organisation recommendations to Listen, Inquire, Validate, Enhance safety and Support (’LIVES’) survivors of DV remain the cornerstone of first-line support (4). Urgently-issued guidelines on safeguarding(5) and responding to DV during COVID-19(6) make a range of recommendations for clinicians supporting people experiencing DV.DV is an important social determinant of physical and mental health, with a range of potential fatal and non-fatal consequences. Despite the constraints of healthcare during a pandemic, attention to patients’ risk of DV and its consequences is a crucial part of bio-psycho-social assessment and management planning. References: (1) Kelly, Morgan. Coronavirus: Domestic abuse calls up 25% since lockdown, charity says. 2020. https://www.bbc.co.uk/news/uk-52157620 (2) Graham-Harrison, et al. Lockdowns around the world bring rise in domestic violence. 2020. https://www.theguardian.com/society/2020/mar/28/lockdowns-world-rise-domestic-violence (3) Roesch, et al. Violence against women during covid-19 pandemic restrictions. BMJ 2020;369. (4) WHO. Responding to intimate partner violence and sexual violence against women: WHO clinical and policy guidelines. 2013. https://apps.who.int/iris/bitstream/handle/10665/85240/9789241548595_eng.pdf;jsessionid=E19DCC3CDAB9BE390EE6F8360C6F1D7E?sequence=1 (5) RCGP. COVID-19 and Safeguarding. 2020. https://elearning.rcgp.org.uk/pluginfile.php/149180/mod_resource/content/2/COVID-19%20and%20Safeguarding%20%286%29.pdf (6) IRISi. Guidance for General Practice teams responding to domestic abuse during telephone and video consultations. 2020. https://irisi.org/wp-content/uploads/2020/04/Guidance-for-General-Practice-Covid-19-FINAL.pdfNo significant relationships.


Acute myeloid leukemia (AML) is a heterogeneous hematologic malignancy with high mortality and poor outcome, especially for elderly/unfit (age ≥ 60 years or unfit patients who are ineligible to receive intensive chemotherapy) with adverse genetic and molecular abnormalities in the newly diagnosed (ND) and refractory/relapsed (R/R) patients.Azacitidine (Aza), a hypomethylating agent, targets epigenetic gene silencing by inhibiting gene expression against malignant phenotypes. The complete remission (CR) rates of Aza combined regimens varied with different drugs, such as, Aza+Midostaurin (20.8%), Aza+Durvalumab 31.3%), Aza+Pracinostat (46%), Aza+Venetoclax (66.4%) in ND AML and Aza+Midostaurin (21.4%), Aza+Nivolumab (22%), Aza+Venetoclax 37.1%) in R/R AML  were enrolled between Jan 2020 and Dec 2021, including 72 ND and 40 R/R Fig. . The 72 2/d on days 1–7 subcutaneous) was given in combination with the HAG regimen . Patients who did not achieve CR/CRi (CR with incomplete blood count recovery) following the first cycle could receive a second cycle at the same doses and schedule. The patients who did not reach CR/CRi after the second cycle were withdrawn from the study. Post-remission therapy for enrolled patients, the Aza+HAG regimen was further given 4–6 cycles or until the disease progresses, two of whom underwent allo-SCT .Induction therapy consisted of Aza . A higher CR/CRi rate of 79.2% in ND AML compared with 40.0% in R/R AML; and sAML (75.0%) obtained a similar high CR rate as de novo AML (80.4%)  achieved CR/CRi within the median time of 33.5d. Notably, 90.4% (66/73) of CR/CRi was obtained after the 1st cycle of Aza+HAG induction therapy. No statistical difference in CR/CRi rate between Aza+HAG (7-day)  vs Aza+HAG (14-day)  (4%) Fig. . In pati4%) Fig. . These dP = 0.0056; median RFS 5.21 m, P = 0.0051)  . In patients with favorable, intermediate, or poor risk, median OS was not reached, 22.8 m, and 17.3 m  and relapse-free survival (RFS) in ND AML were 22.8 m and not reached, respectively, which were longer than R/R AML  Fig. . De novo48) Fig. . In pati3 m Fig. ; median 3 m Fig. , respectP = 0.016) , respectively . The baseline demographic and characteristics were generally balanced between the two cohorts Table . The CR/16) Fig. . The medely Fig. . We alsoely Fig.  and S2. The most common non-hematological adverse event (AE) was infection (58.0%). Common non-hematological AEs of grade 3 or higher includes: infection (33.9%), hemorrhage (9.82%), fatigue (5.36%), hypokalemia (3.57%), cardiac arrhythmia (1.79%), and fever (1.79%). For the hematological AE, the median duration of neutropenia and thrombocytopenia were 11 d  and 16d , 10 d , and 16 d , 12d  and 17d  for total, 7-day, and 14-day schedule, respectively. No differences were observed between the 7-day and 14-day schedule. The early deaths within 4 weeks of the induction treatment occurred in 1.79%. No patients discontinued the induction therapy due to hematological or non-hematological toxicities Table . These dDNMT3A, TET2, and NPM1 , KIT , IDH1 , NPM1 , ASXL1 , DNMT3A , RUNX1 , FLT3  and TET2   and 87.4% (90/103) of patients had more than one gene mutation. The most frequently mutated genes were PM1 Fig. . We obse14) Fig.  (Table S14) Fig. . We alsoPM1 Fig. . OS and ion Fig. ; whereasype Fig. . These dIDH1 inhibitor Ivosidenib (60.9%) [IDH1-mutant ND AML or HMA + Venetoclax (71%) in IDH1/2-mutant AML [IDH1-mutant AML and the IDH1 mutant was dramatically reduced after the first cycle of Aza+HAG, suggesting that the patients with the IDH1 mutation could achieve a remarkable deep and durable remission upon Aza+HAG treatment.Recent studies reported the CR rates of patients treated with the Aza plus  (60.9%)  in IDH1-tant AML . This trIn summary, this trial demonstrated that the Aza+HAG regimen is a cost-effective first-line therapy with high efficacy and well tolerance for elderly/unfit AML. This trial provides a rationale for further expanding the patients for randomized clinical controlled studies and guiding suggestions for the clinical use of this novel combination therapy.Supplemental methods_tables_figures


Background and Objectives: To estimate the association between admission functional outcomes and exposure to physiotherapy interventions with mortality rate in intensive care unit (ICU) inpatients with cardiovascular diseases and new coronavirus disease (COVID-19). Materials and Methods: Retrospective cohort including 100 ICU inpatients (mean (standard deviation), age 75 (16) years) split into COVID-19+ or COVID-19−. The association of in-ICU death with admission functional outcomes and physiotherapy interventions was investigated using univariable and multivariable regression models. Results: In total, 42 (42%) patients tested positive for COVID-19. In-ICU mortality rate was 37%, being higher for the COVID-19+ group : 3.15 (1.37–7.47), p = 0.008). In-ICU death was associated with lower admission ICU Mobility Scale score (0.81 (0.71–0.91), p = 0.001). Restricted mobility (24.90 (6.77–161.94), p < 0.001) and passive kinesiotherapy (30.67 (9.49–139.52), p < 0.001) were associated with in-ICU death, whereas active kinesiotherapy (0.13 (0.05–0.32), p < 0.001), standing (0.12 (0.05–0.30), p < 0.001), or walking (0.10 (0.03–0.27), p < 0.001) were associated with in-ICU discharge. Conclusions: In-ICU mortality was higher for inpatients with cardiovascular diseases who had COVID-19+, were exposed to invasive mechanical ventilation, or presented with low admission mobility scores. Restricted mobility or passive kinesiotherapy were associated with in-ICU death, whereas active mobilizations  were associated with in-ICU discharge in this population. Cardiovascular diseases (CVDs) are the leading cause of death globally, with estimated 31% mortality, representing about 17.9 million deaths every year . The BraThe hazards of hospitalization, particularly for older adults in the intensive care units (ICU), are a longstanding issue that ultimately favors a decline in musculoskeletal function and functional capacity . Higher Retrospective, single-center study. Data were obtained by the principal investigator through information previously contained in electronic medical records, examination reports, and notes of the health professional staff involved in the care of the patients. This study is reported following the REporting of studies Conducted using Observational Routinely-collected health Data (RECORD) statement . MinimumThis study retrospectively analyzed all data from February to November 2020 collected from patients consecutively hospitalized at the ICU of a primary-to-tertiary private hospital located in Curitiba, Paraná, Brazil.ICU admission criteria comprised at least of the following conditions: hemodynamic ; neurological ; or respiratory . Other causes for ICU admission included postoperative period of high-risk patients, postoperative period of large surgeries, postoperative cardiac surgery, postoperative neurological surgery, postoperative endovascular surgery, need for clinical monitoring, sepsis, septic shock, cardiac arrhythmia, vascular diseases of the heart, or acute renal dysfunction.Patients who had a primary diagnosis of CVD after a complete clinical exam and laboratory testing including laboratory blood tests, electrocardiogram, blood pressure, and/or echocardiography as prescribed, admission assessment by a physiotherapist, and tested for SARS-CoV-2 infection at admission were included. ICU admission was defined as an admission to the hospital’s ICU for >12 h. Re-admissions of patients to the ICU within the study period were excluded from the analysis.All admission data were collected within <24 h of ICU hospitalization at the discretion of the medical staff and covered the required time for swab analysis. Data were collected retrospectively from electronic medical recordings regarding demographics, vital signs, laboratory, gasometry, presence of CVD and comorbidities, and drugs in continuous use. Date of hospital admission and discharge from the ICU or death were collected for computing the total length of ICU stay. The sample was divided into groups COVID-19+ or COVID-19− based on the ICU admission test, after a nasal and/or nasopharyngeal swab for SARS-CoV-2 by polymerase chain reaction method.Overall muscle strength was assessed by the Medical Research Council (MRC) scale, which uses a 6-point scale of 6 muscle groups bilaterally. Representative scores comprised the sum of points observed for each muscle group bilaterally, ranging from 0 (no muscle activity) to 60  .Mobility was assessed by the ICU Mobility Scale (IMS). The score varies between 0 expressing low mobility (patient who only performs passive exercises in bed) and 10 expressing high mobility  .All patients were exposed to physiotherapy interventions based on the hospital standards of usual care developed according to international and national recommendations ,27,28,292O and subsequent titration of ideal PEEP, provided they were clinically stable. When they needed oxygen therapy, it was performed using a low-flow system . Spontaneous prone was also used for at least 1 h. In the supine position [Ventilatory support was characterized using non-invasive mechanical ventilation, through an orofacial or facial interface connected to the mechanical ventilator in one or two pressure levels ventilation modes, or invasive ventilatory support . Patients diagnosed with acute respiratory distress syndrome used protective strategy ventilatory parameters, which may require alveolar recruitment through the prone position or recruitment through the gradual increase of positive end-expiratory pressure (PEEP) up to 35 cmHposition , the heaMobility activities were categorized as complete bed restriction; passive kinesiotherapy ; active kinesiotherapy ; assisted or active sitting out of bed; standing; and walking.The primary outcome was in-ICU mortality as well as admission functional assessments of MRC and IMS scores. In-ICU mortality was calculated from the admission date and confirmed using electronic medical records.No missing data occurred for exposures, in-ICU mortality, or admission IMS scores. Data were missing for the admission assessment of MRC scores in 18/100 participants due to sedation.p < 0.05 (two-tailed).Statistical analysis was performed in jamovi v. 1.8.1.0 and R project version 4.0.4 with packages after importing the electronic spreadsheet. Missing data in admission measurements were reported and assumed to be missing completely at random and univariate mean imputation was performed. Evidence of statistical significance was considered at Descriptive summaries were reported as mean (standard deviation (SD)) for continuous variables or absolute and relative frequencies (%) for categorical ones. Admission demographic data were compared between COVID-19+ versus COVID-19− groups using the linear model analysis of variance or Pearson’s Chi-squared test for continuous and dichotomous variables, respectively.Univariable logistic regression analysis was performed to examine the association (odds ratio (OR) with 95% confidence interval (95% CI)) of exposure to physiotherapy intervention (ventilatory support and mobility) with group (COVID-19+ vs. COVID-19−) and in-ICU mortality. Multivariable logistic regression model was fitted to determine independent factors associated with in-ICU mortality; all factors related to exposure to physical therapy were force-entered as a full model. Model fit was evaluated by Akaike information criterion (AIC) and C-statistic.2). The most common underlying CVD was hypertension (91/100), followed by a history of cerebrovascular disease (22/100), coronary artery disease (21/100), heart failure (16/100), and atrial fibrillation. Overall length of ICU stay was 9.7 (15.5) days, with patients with COVID-19+ showing longer ICU stay (14.5 (21.7) vs. 6.2 (6.8), p = 0.007). Between-groups comparisons show that patients with COVID-19+ showed higher admission MRC scores (48.3 (7.9) vs. 43.8 (10.4), p = 0.021) and IMS scores (5.5 (4.0) vs. 3.9 (3.8), p = 0.039). They were also younger (68 (16) vs. 80 (13) years, p < 0.001) and presented at admission with lower leukocytes count (9565 (5473) vs. 13,456 (6444) count/mcL, p = 0.002), lower partial pressure of carbon dioxide (PCO2) (31.7 (6.5) vs. 37.7 (8.5), p < 0.001), and lower bicarbonate (21.9 (4.5) vs. 23.8 (4.9) mEq/L, p = 0.044). Sedation at admission was more frequent in the COVID-19+ group . No statistical evidence of difference was observed between groups for severity of disease based on acute physiology and chronic health evaluation (APACHE II: 30.3 (4.8) vs. 30.2 (4.9), p = 0.917).Overall, the sample was composed of older adults (75 (16) years), balanced between sexes , and with most participants (40/100) classified as overweight (body mass index = 27.1 (5.1) kg/mp = 0.011) or prone position (22.80 (4.19–425.35), p = 0.003), to alveolar recruitment (19.04 (4.97–125.99), p < 0.001), awake prone (5.60 (1.27–39.05), p = 0.038), or length of stay (1.07 (1.02–1.13), p = 0.008), but not to non-invasive mechanical ventilation or oxygen therapy. Multivariable logistic regression analysis showed a good linear fit (C-statistic = 0.783), with COVID-19 remaining independently associated with exposure to alveolar recruitment (22.34 (3.56–224.91), p = 0.002) or awake prone (13.41 (1.62–228.22), p = 0.032).As related to ventilatory support , patientp = 0.029).As related to mobility during ICU stay , no statp = 0.008). As related to ventilatory support, in-ICU mortality was more likely in patients exposed to invasive mechanical ventilation in either supine (22.71 (8.28–70.76), p < 0.001) or prone position (4.74 (1.42–18.74), p = 0.016), oxygen therapy (8.75 (2.34–57.11), p = 0.005) or alveolar recruitment , p < 0.001). Also, in-ICU death was associated with longer length of stay (1.04 (1.01–1.09), p = 0.048)), lower admission IMS score (0.81 (0.71–0.91), p = 0.001) but not MRC score (p = 0.055). Multivariable logistic regression analysis showed an excellent linear fit (C-statistic = 0.920), with in-ICU death still independently associated with COVID-19+ , exposure to invasive mechanical ventilation (14.81 (2.97–97.70), p = 0.002) and admission IMS score (0.79 (0.63–0.96), p = 0.023).p < 0.001), passive kinesiotherapy (30.67 (9.49–139.52), p < 0.001) and longer LOS (1.04 (1.01–1.09), p = 0.048) were associated with in-ICU death, whereas active kinesiotherapy (0.13 (0.05–0.32), p < 0.001), standing (0.12 (0.05–0.30), p < 0.001), or walking (0.10 (0.03–0.27), p < 0.001) were associated with in-ICU discharge. Multivariable logistic regression analysis showed an excellent linear fit (C-statistic = 0.947), with in-ICU death still independently associated with COVID-19+ (15.44 (2.80–140.82), p = 0.005), restricted mobility (10.49 (1.74–95.85), p = 0.017), and passive kinesiotherapy (17.66 (2.32–326.56), p = 0.017).As related to mobility during ICU stay, two patterns emerged: restricted mobility (24.90 (6.77–161.94), The main findings suggest that in-ICU mortality is higher for inpatients with CVD who had a COVID-19+ test result, were exposed to invasive mechanical ventilation, or presented with low admission IMS scores, whereas restricted mobility or passive kinesiotherapy were associated with in-ICU death, active mobilizations  were associated with in-ICU discharge. Our findings contribute to the global discussion on the acute management of patients with COVID-19 by providing insights about the planning physiotherapy interventions aimed at reducing in-ICU fatality among patients with CVD.Demographic and clinical data from our sample in Brazil corroborate previous studies in other countries on the risk factors for hospitalization and mortality in patients with CVD and COVID-19: mainly older age, overweight, low lymphocyte count, and pre-existing comorbidities, among others ,9,31,32.Clinical algorithms  and consIn-ICU mortality was higher for inpatients with CVD who had a COVID-19+ test result, were exposed to invasive mechanical ventilation, or presented with a lower admission IMS score. Altogether, these characteristics can be understood as a proxy for disease severity. Interestingly, exposure to physiotherapy intervention showed two distinct effects on in-ICU mortality rate, whereas restricted mobility or passive kinesiotherapy were associated with in-ICU death, active mobilizations  were associated with in-ICU discharge. This finding corroborates previous studies showing improved mobility at hospital discharge and a higher probability of discharging home with increased frequency and longer mean duration of physical therapy visits in patients with COVID-19 admitted to acute care hospitals . ConsideThis study has some limitations. Due to the retrospective design, there were missing data for participants regarding admission assessment of functional outcomes. Clinical data at admission were collected within <24 h of ICU hospitalization and hence may differ from pre-admission status that required hospitalization. Moreover, physiotherapy interventions were delivered as per the rehabilitation team’s clinical decision-making process. Whereas such lack of control in experimental factors may have influenced the delivered interventions in each group, such pragmatic approach most likely represents ICU routines in Brazil since they are based on national guidelines. Some CIs returned wide ranges, which suggests a large uncertainty about the effect  and that further information is needed. Nonetheless, assessment of goodness-of-fit of the models (Akaike information criterion and C-statistics) suggest acceptable model validity. The current sample is from a single center during the first ‘wave’ (February to November 2020) of cases in Brazil  when varIn-ICU mortality is higher for inpatients with CVD who had COVID-19+, were exposed to invasive mechanical ventilation, or presented with low admission mobility scores. The protective effects of routine physiotherapy interventions are highest when patients can perform active rather than passive kinesiotherapy. Patients with COVID-19 who further perform standing and walking activities appear to experience a higher survival effect.


Tuberculosis (TB) remains a leading infectious cause of death and morbidity globally. Rifampin-resistance or intolerance requires prolonged treatment using less effective, more toxic regimens. Recent trials demonstrated that the all-oral six-month “BPaL” regimen, Bedaquiline, Pretomanid and Linezolid, is 90% effective. In these trials, linezolid-induced hematologic and neurologic toxicity was high using 1200mg daily, whereas lower exposure (dose or duration) reduced toxicity. Therapeutic drug monitoring (TDM) is used by U.S. TB experts to maintain a serum linezolid trough < 2ug/ml, which correlates with reduced toxicity. Since U.S. FDA approval in 2019, BPaL has been widely implemented for the treatment of rifampin-intolerant or resistant TB disease.We evaluated a cohort of patients with TB treated from October, 2019 through April, 2022, describing patient demographics, BPaL treatment dosing, and adverse events. TDM was performed for clinical purposes using liquid chromatography–mass spectrometry to measure serum levels at trough and 2 and 6h post-linezolid dose. Clinical providers adjusted linezolid dose and dosing interval targeting a trough < 2ug/ml and peak of 12–26ug/ml.Among 64 BPaL patients, ages were 15–83 years, 22 (34.3%) were female, 6 (9.3%) U.S.-born, 4 (6.3%) HIV-infected. 50 (78.1%) had only pulmonary disease, 6 extrapulmonary, and 8 had both; 61 (91.0%) were culture-confirmed. Most (n=62) started linezolid 600mg daily. Linezolid was adjusted for 39 (66.1%) of the 59 patients with TDM; 18 had a trough >2ug/ml, 30 had dosing interval increased to thrice-weekly, and 17 had a dose increase. 52 (81.3%) patients completed BPaL and 12 remain on therapy. One 81-year-old female with diabetes, hypothyroidism, and B12 deficiency discontinued linezolid at 12 weeks for worsened neuropathy (linezolid trough=1.13ug/ml). She completed 26 weeks of bedaquiline/pretomanid and her symptoms returned to baseline.Use of BPaL with clinical and TDM monitoring has transformed treatment of rifampin-resistant or intolerant TB in the U.S. Patients previously sentenced to 18–24 months of treatment with 5–7 hard-to-tolerate medications and modest efficacy can now complete treatment in 6–9 months with little toxicity and exceptional cure rates.All Authors: No reported disclosures.


Nocardia farcinica is an opportunistic pathogen that causes nocardiosis primarily in patients with compromised immune systems. In this study, we used the genetically tractable organism Caenorhabditis elegans as a model to study the innate immune responses to N. farcinica infection. We found that unlike other pathogenic bacteria such as Pseudomonas aeruginosa and Staphylococcus aureus, N. farcinica failed to kill adult worms. In another words, adult worms exposed to N. farcinica exhibited a normal lifespan, compared with those fed the standard laboratory food bacterium Escherichia coli OP50. Interestingly, deletion of three core genes  in the p38 MAPK/PMK-1 pathway reduced the survival of worm exposure to N. farcinica, highlighting a crucial role of this pathway for C. elegans in resistance to N. farcinica. Furthermore, our results revealed that N. farcinica exposure up-regulated the level of PMK-1 phosphorylation. The activation of PMK-1 promoted nuclear translocation of a transcription factor SKN-1/Nrf2, which in turn mediated N. farcinica infection resistance in C. elegans. Our results provide an excellent example that the integrity of immune system is key aspect for counteract with pathogenesis of N. farcinica. Nocardia farcinica, is an opportunistic pathogen under the genus of Actinomycetes widely distributed in the soil and rotten substances, causing lung infection through inhalation of bacterial particles [N. farcinica, suppress immune responses by inhibiting Notch signal pathway in RAW264.7 macrophages [N. farcinica in mammal. The mechanism of pathogenesis in N. farcinica infection is not fully understood.articles . Since tarticles . Nocardiarticles . Nocardiarticles . A recenCaenorhabditis elegans is a free-living, bacterivorous nematode that lives in the soil, well known as invertebrate without adaptive immune system as well as specific immune cells [C. elegans often encounters a variety of pathogens, thus it exerts a strong selective pressure to evolve and develop an effective innate immune system. C. elegans therefore represents a powerful invertebrate model to study the innate immune responses to nosocomial bacterial pathogens such as Pseudomonas aeruginosa, Staphylococcus aureus, Salmonella enterica, Salmonella typhimurium, and Serratia marcescens [Cryptococcus neoformans [Candida albicans [ne cells . In wildne cells . In the rcescens ,8,9,10, oformans  and Candalbicans .C. elegans have revealed a variety of the evolutionarily conserved signaling pathways in the innate immunity, such as the DAF-2 insulin-like signaling pathway [C. elegans to regulate innate immunity [C. elegans upon P. aeruginosa infection [P. aeruginosa infection [During the last two decade, studies using  pathway , the p38 pathway , and the pathway , the JNK pathway , the pro pathway , the G p pathway , the G p pathway , and the pathway . Of thesimmunity ,21. PMK-immunity . Meanwhinfection . Interesnfection . Under nnfection .N. farcinica in C. elegans. We discovered that the PMK-1/p38 MAPK signaling was required for the survival of worms upon N. farcinica exposure. Further studies indicated that PMK-1/p38 MAPK activated the transcription factor SKN-1, which in turn conferred resistance to N. farcinica infection in C. elegans.In this study, we screened the major signaling pathways that are involved in defense against N. farcinica in nematodes. However, we found that unlike worms exposed to P. aeruginosa, S. aureus, and E. faecium [N. farcinica exhibited a similar lifespan as those fed E. coli OP50, the standard laboratory food . In contrast, pmk-1(km25) mutants exhibited marked decrease in lifespan when exposed to N. farcinica, compared to worms fed on E. coli OP50 and WT (nsy-1(ag3) and sek-1(ag1) mutants exposed to N. farcinica also showed a significant reduction in lifespan, compared to E. coli OP50 diet and WT , sek-1(ag1), and pmk-1(km25) suppressed their mRNA levels , catalase (katG), and superoxide dismutase (sodF) are predicted with high expressivity in N. farcinica, which could defense against reactive oxygen species (ROS) [N. farcinica infection. Thus, ROS are unlikely to be involved in the activation of the p38 MAPK pathway in worms.We then asked if oli OP50 a,b. The osa PA14  and Cutium acnes . A previby PMK-1 . We thuspression c,d. As Fed genes , we thenA levels e. Taken es (ROS) . We thusC. elegans, SKN-1 confers resistance to Gram-negative bacteria P. aeruginosa PA14 and Gram-positive bacteria E. faecalis [N. farcinica infection, we used transgenic worms expressing SKN-1b/c::GFP+rol-6 to detect the nuclear translocation of SKN-1, which is an indicator of its activation [N. farcinica exposure. However, knockdown of nsy-1, sek-1 or pmk-1 by RNAi significantly reduced the nuclear translocation of SKN-1 in worms exposed to N. farcinica (gst-4 (encoding Glutathione S-transferase-4) is a target gene of SKN-1 [Pgst-4::gfp reporter gene to further demonstrate the activation of SKN-1 by N. farcinica infection. We found that the expression level of Pgst-4::gfp was dramatically increased in worms exposed to N. farcinica for 24 hours, The increase is abolished when worms subjected to skn-1, nsy-1, sek-1 or pmk-1 RNAi (skn-1(tm3411), nsy-1(ag3), sek-1(ag1), and pmk-1(km25) suppressed up-regulation of gst-4 expression induced by N. farcinica infection  mutants was comparable to that of the pmk-1(km25) mutant subjected to skn-1 RNAi upon N. farcinica infection  of N. farcinica and E. coli OP50 in the body of worms  worms, skn-1 (RNAi) worms, and control group worms. As black arrows pointing out, genetic inactivation of pmk-1 and skn-1 caused necrosis in the head of worms exposed to N. farcinica, but not to E. coli OP50 , known as key immune factor, mediates resistance to Mycobacterium. spp [N. farcinica in organisms ranging from C. elegans to mammals.In ruginosa ,38 S. au. aureus , Salmonelmonella , Yersinia pestis , and Mycbacteria . The p38bacteria  secreted O157:H7 . In mamm O157:H7 ,44, suggium. spp ,46. ThesP. aeruginosa, E. faecalis [Aeromonas dhakensis [N. farcinica infection. It has been reported that PMK-1 can phosphorylate SKN-1 at Ser-74 and Ser-340 [P. aeruginosa, E. faecalis and N. farcinica infection activates SKN-1 in a p38 MAPK-dependent manner. These results suggest that SKN-1 is one of downstream effectors of p38 MAPK that mediates C. elegans innate immune responses to both Gram-negative and Gram-positive pathogens. The molecular mechanism underlying SKN-1-mediated innate immunity remains unclear. Previously, we and others have demonstrated that autophagy plays an important role in C. elegans defense against a variety of pathogenic bacteria by repairing organismal insults [glp-1(e2141ts) mutants [hyl-1; lagr-1 mutants [N. farcinica infection via promoting autophagy. The opportunistic infections followed immunologic deficiency have mainly been seen in HIV infected patients, revealing that the environmental “harmless” microorganisms could be life threaten, such as Penicillium marneffei, known as the third common opportunistic pathogen that causes penicilliosis in HIV patients. Likewise, N. farcinica is an opportunistic pathogen, with the majority of infections occurring in immunocompromised patients. In the current study, WT worms do not have a reduced lifespan when exposed to N. farcinica. These data suggested that like humans, WT worms with an intact innate immune system can resist to N. farcinica infection, but lacking key genes in the PMK-1/p38 MAPK pathway become susceptible to N. farcinica infection. In our studies, we also noticed that lack of p38 MAPK pathway and SKN-1 leads to significant increases accumulation of N. farcinica and obvious enlarged vacuoles in the head of worms. Thus, our finding emphasizes that the integrity of immune system is crucial for defense against N. farcinica infection in C. elegans.As a transcription factor, SKN-1 is involved in resistance to oxidative stress by upregulating a set of the Phase II detoxification genes . SKN-1 afaecalis , Mycobacfaecalis , and Aerhakensis . In the  insults ,20,48. S mutants . Meanwhi mutants . Thus, SN. farcinica to activate the p38 MAPK/PMK-1 pathway?However, our data demonstrate that ROS are not the signal component produced by N. farcinica. Several virulence factors of N. farcinica have been reported when the pathogen infected mammals. For instance, Nfa34810 protein of N. farcinica can stimulate macrophages to produce tumor necrosis factor alpha (TNF-α) and other immune related factors through the TLR4 pathway [N. farcinica IFM 11523, which isolated from Japanese patient, secretes two pathogenesis factors nocobactin NA-a (compound 1) and nocobactin NA-b (compound 2), which act as on the notch signaling inhibitors [What are the critical signal molecules from  pathway . BesidesC. elegans strains were maintained on NGM medium containing E. coli OP50 under standard condition. Strain used in this study include C. elegans Bristol N2, pmk-1(km25), mpk-1(n2521), let-60(ga89), jnk-1(gk7), hlh-30(tm1978), skn-1(tm3411), nsy-1(ag3), sek-1(ag1), skn-1b/c::gfp+rol-6(su1006), nlp-29p::gfp+col-12p::DsRed, gst-4p::gfp::NLS(pAF15) were kindly provided by the Caenorhabditis Genetics Center (CGC), which is funded by NIH Office of Research Infrastructure Programs (P40 OD010440). Bristol N2 strain of C. elegans is used as non-genetic modified control in experiments, and labeled as wild-type (WT) in our study [The , mpk-1n21, let-60ur study .E. coli HT115 (DE3) strain carrying vector L4440 express dsRNA corresponding to targeted genes to perform double-stranded (ds)RNA-mediated RNA interference (RNAi). As experimental control, E. coli HT115 carrying empty vector (no dsRNA expression) were used, labelled as EV. All strains were grown overnight in LB medium with 100 μg/mL ampicillin at 37 °C. Cultured E. coli strains spread onto NGM plates contained 100 μg/mL ampicillin and 1 mM isopropyl 1-thio-β-D-galactopyranoside (IPTG). RNAi assay were performed according to pervious study [2 strain of C. elegans) fed on RNAi bacteria strains at 20 °C until reach to young adult. Then young adult worms were transferred on the BHI plates for further studies.Bacterial strains contain RNAi targeting genes were obtained from Ahringer library . E. colius study , in brieC. elegans strains were cultured on NGM plate with E. coli OP50 at 20 °C following the standard maintenance procedures [E. coli OP50 for further assays. To examine the effect of N. farcinica on worms’ lifespan, synchronized L1 larvae were cultivated fed E. coli OP50 at 20 °C until the young adult stage. Then 50–60 worms were fed with either N. farcinica or laboratory standard diet E. coli OP50 on BHI medium containing 5′-fluoro- 2′-deoxyuridine (FUdR) (75 μg/mL) at 25 °C. The survival curves were plotted by scoring the dead worm at 24 h interval. Immobile worms unresponsive to touch were scored as dead. Three plates were analyzed per assay and all experiments were performed three times. The ocedures . Worms wocedures , eggs weskn-1b/c::gfp were fed with either N. farcinica or laboratory standard diet E. coli OP50 on BHI plates for 24 hours. Briefly, worms were mounted on slides from plates and imaged by using Nikon e800 fluorescence microscope. At least 30 worms were examined under each condition in three independent experiments.After young adult worms expressing After washed with M9 buffer, worms were homogenized in liquid nitrogen. Then the homogenate was lysed on ice for 30 min in lysis buffer RIPA . After centrifuged at 12,000 rpm for 15 min at 4 °C, the supernatant was obtained and used for Western blot analysis. The total protein extraction was loaded on 10% SDS-PAGE for electrophoresis. Proteins were then transferred to immobilon-PSQ transfer PVDF membrane . Primary antibodies were anti-phospho-p38 antibodies , and anti-α-tubulin antibodies . The secondary antibodies were peroxidase-coupled anti-rabbit IgG . Blots were developed using Super Signal chemiluminescence substrate . An imaging system (Amersham Imager 600) was used for documentation of the Western blot results. Band intensities were measured using ImageJ software (NIH).® Premix-Ex TagTM  on a Roche LightCycler 480® System . act-1 gene was used for an internal control. The primers used for PCR are listed in Total RNA was isolated from worms with TRIzol Reagent . Random-primed cDNAs were generated by reverse transcription of the total RNA samples with SuperScript II (Invitrogen). A real time-PCR analysis was conducted using SYBRnlp-29p::gfp or gst-4p::gfp were exposed to N. farcinica or E. coli OP50 for 24 h. Then the worms were mounted in M9 onto microscope slides. The slides were imaged using a Nikon e800 fluorescence microscope. Fluorescence intensity was quantified by using the ImageJ software (NIH). Mean value and standard errors were calculated based on more than 30 worms under each condition in three independent experiments.For detecting fluorescence in worms, analysis, synchronized young adult worms expressing either ROS was detected by transferring worms in to M9 buffer with DHE (3 μM) and stained for 3 h before mounting in M9 buffer onto microscope slide, examined by fluorescence microscope (Zeiss Axioskop 2 Plus) .N. farcinica colonized in worms, we fed young adult worms with either E. coli OP50 or N. farcinica for 96 h on BHI plates, then worms were transferred in to M9 buffer containing 25 mM levamisole hydrochloride , 50 μg/mL kanamycin and 100 μg/mL carbenicillin , and soaking for 30 min before washing with M9 buffer for three times. Worms were collected and grinded in PBS with 0.1% Triton, and serial diluted before placing on BHI ager for incubation, then bacterial colonies were counted to measure CFU.To ask if t-test. Differences in survival rates were analyzed using the log-rank test. Difference of CFU counting was assessed by two-tailed unpaired t-test. Data were analyzed using SPSSsoftware, version 26.0  and Graphpad Prism 8.The statistical significance of differences in gene expression and fluorescence intensity was assessed by performing a one-way ANOVA followed by a Student-Newman-Keuls test and two-tailed unpaired


Endoscopic Endonasal Surgery (EES) is an innovative surgical technique to remove brain tumors and lesions. Post-operative central nervous system (CNS) infections following EES are poorly described. The objective of this study was to define the epidemiology and characteristics of post-EES CNS infections.Adult patients who underwent EES between 1/2010 and 7/2021 were evaluated and included if microbiologically confirmed CNS infection occurred within 30 days of EES. Suspected contaminants, ventricular drain colonization, and pre-EES CNS infections were excluded.S. aureus, Enterobacterales, and P. aeruginosa. Among 20 patients with prior EES, pathogens included S. aureus (5/20), Enterobacterales (3/20), Enterococcus spp. (3/20) and polymicrobic infections (3/20). Overall, 35.1% (13/37) of patients developed CNS infection due to a pathogen susceptible to pre-EES prophylaxis. Among those colonized with MRSA at time of EES, 75% (3/4) developed MRSA CNS infection compared to 6.1% (2/33) of non-colonized MRSA patients (P=0.005). The overall 30-day mortality rate was 2.7% (1/37).Overall, 2005 patients underwent EES; 1.8% (37/2005) developed CNS infection. The median [IQR] age was 51 [42-60] years, 32.4% (12/37) were female, and 54% (20/37) had a prior EES. The most common indications for EES were tumor resection [67.6% (25/37)] and cerebrospinal fluid (CSF) leak repair [24.3% (9/37)]. Post-operative CSF leaks were documented in 70.3% (26/37) of patients and 24.3% (9/37) had an extra-ventricular drain or shunt in place for >48 hours at the time of infection. Ceftriaxone prophylaxis was prescribed in 64.9% (24/37) of cases and other regimens varied. The median [IQR] time from EES to diagnosis of CNS infection was 12 [6-19] days. The most common pathogens were A polymicrobic case was defined as >1 pathogen isolated from CSF (n=1) or from rhinocerebral tissue if CSF cultures were negative (n=11). Among polymicrobic cases (n=12), P. aeruginosa (n=5), Enterococcus spp. (n=4). and S. aureus (n=3) were predominant. Cases labeled as other consisted of Trichoderma spp, A. xylosoxidans, P. acnes, S. epidermidis, Peptostreptococcus spp.S. aureus, antimicrobial prophylaxis should ensure adequate coverage of this pathogen in addition to sinus flora, and programs may benefit from screening patients for MRSA colonization pre-EES. Our data also suggest that prophylaxis should target Gram-negative and other colonizing bacteria among patients with prior EES.CNS infection post-EES is rare and causative pathogens vary. Given the predominance of Ryan K. Shields, PharmD, MS, Infectious Disease Connect: Advisor/Consultant|Merck: Advisor/Consultant|Merck: Grant/Research Support|Roche: Grant/Research Support.


Attention-Deficit/Hyperactivity Disorder (ADHD) is associated with alterations in both reinforcement sensitivity and affective processing but the nature of the associations of these characteristics is yet to be examined. We hypothesized that individual differences in the sensitivity of the Behavioral Approach System (BAS) would exhibit differential relations with affective network connectivity – involved in emotional regulation and salience monitoring – in youth at-risk for, relative to youth not at-risk for, ADHD.n = 125; Mage=16.24 years, SD = 1.09 years; 61.6% boys) were recruited as part of The Budapest Longitudinal Study of ADHD and Externalizing Disorders. Forty-nine were classified as at-risk for ADHD , defined as exhibiting ≥4 parent-rated symptoms of either domain on the ADHD Rating Scale-5. Participants completed a 10-minute resting-state functional Magnetic Resonance Imaging session, during which they were asked to focus their attention on a fixation cross, as well as various self-report assessments, including the Reinforcement Sensitivity Theory of Personality Questionnaire (RST-PQ).Adolescents  within a cluster based on functional similarity . Follow-up OLS linear regressions showed higher impulsivity scores predicted stronger functional connectivity between the (1) left amygdala-right insula  = 3.298, p = .005, adjusted R2=.101), (2) left amygdala-left insula  = 2.2, p = .048, adjusted R2=.055), (3) right amygdala-right insula  = 3.833, p = .002, adjusted R2=.121), and (4) right amygdala-left insula  = 3.064, p = .008, adjusted R2=.092) in at-risk youth, whereas an inverse relationship was apparent in not at-risk youth.Functional Network Connectivity analyses indicated an interaction effect between the RST-PQ BAS impulsivity subscale and at-risk status on functional connectivity between four affective network region-pairs (t(122)=−1.167, p = .246) or on functional connectivity ((1) t(122) = .383, p = .702; (2) t(122) = .195, p = .846; (3) t(122)=−.107, p = .915; (4) t(122)=−.206, p = .837).There was no main effect of group status on BAS impulsivity scores  as the role of the amygdala-insula connection in reward sensitivity appears especially relevant for a developmental phase and a diagnostic group linked to increased risk taking.


Relative transformation efficiencies ranged from − 44 to + 45% compared to Silwet® L-77. Surfactants S200, S240, and S279 demonstrated the greatest enhancement in transformation.Floral dip transformation of Arabidopsis has been used consistently for 20 years with little change in the protocol. Here we directly compare seven novel surfactants (BREAK-THRUThe online version contains supplementary material available at 10.1186/s13104-022-06251-5. Plant biotechnology offers tremendous promise for improving crops by increasing the sustainability and reducing the environmental impact of production. These technologies have led to crop plants with enhanced nutritional properties, abiotic stress tolerance, disease and pest resistance, and other agronomic traits, as well as crop tools for accelerated precision breeding. Although genetic engineering and genomic editing approaches have been applied successfully to various plant species, relatively few crops have an easy and efficient means of transformation, which restricts progress in research as well as agricultural deployment. Thus, additional techniques for broadly and efficiently applying transformation technologies to a wide range of crops and their constituent elite varieties are required.One of the earliest enhancements to transformation was the use of surfactants, which modify the surface properties of liquids to enhance their spreading, wetting, emulsifying, dispersing, or other characteristics. Surfactants have been used extensively in herbicide and pesticide formulations to increase their penetration. However, they must be used judicially, as surfactants may have inhibitory phytotoxic effects on plant tissue at high concentrations and stimulatory effects at low concentrations , 2.Agrobacterium-mediated transformation of Arabidopsis thaliana  in water: 25 (0.1%).BREAK THRU®-S200: Trisiloxane super-spreader (more water-soluble), stable within pH range 6–8, Static surface tension [mN/m] in water: 22 (0.1%).BREAK THRU®-S233: Trisiloxane super-penetrant  systemic products, stable within pH range 6–8, Static surface tension [mN/m] in water: 23 (0.1%).BREAK THRU®-S240: Trisiloxane super-spreader (liquid soluble), stable within pH range 6–8, Static surface tension [mN/m] in water: 22 (0.1%).BREAK THRU®-S279: Trisiloxane super-spreader (more oil-soluble), stable within pH range 6–8, Static surface tension [mN/m] in water: 21 (0.1%).BREAK THRUS301: Trisiloxane super-spreader, stable within pH range 6–8, Static surface tension [mN/m] in water: 22 (0.1%).®-SP133: Polyglycerol ester-based adjuvant , stable within pH range 4–9, Static surface tension [mN/m] in water: 29 (0.1%).BREAK THRU® compound see (www.evonik.com/break-thru) [For more details of the BREAK THRUak-thru) .We show an increase in transformation efficiency for Arabidopsis floral dip transformation using a published technique . HoweverAdditional file 1. Statistics 1: Figure S1.Additional file 2. PCR: Figure S2.Additional file 3. Statistics 2: Table S1. Surfactant transformation efficiency. Table S2. Surfactant transformation statistics.


Nature Communications 10.1038/s41467-021-25159-5, published online 09 August 2021Correction to: Nat. Commun. 12, 1–14 (2021). The correct reference for Ref. 11 is: Zhang, Y. et al. mTORC1 couples cyst(e)ine availability with GPX4 protein synthesis and ferroptosis regulation. Nat. Commun. 12, 1589 (2021). This has been corrected in the PDF and HTML versions of the Article.The original version of this Article contained an error in Ref. 11, which incorrectly gave the reference as: Zhang, Y. et al. mTORC1 couples cyst(e)ine availability with GPX4 protein synthesis and ferroptosis regulation.


E.coli (STEC) infection which presents a characteristic triad of microangiopathic hemolytic anemia, thrombocytopenia, and acute renal failure. Preliminary in vitro and experimental animal studies demonstrated that Shigatoxins (STXs) induce the secretion of proinflammatory cytokines. Human neutrophil gelatinase-associated protein  has been reported as a marker of acute kidney injury. Dosing serum levels of IL-8, TNF-α, IL-6, IL-1β and N-gal in children with STEC-associated infection and HUS would allow to establish the role of these cytokines as biomarkers of renal injury and severityHemolytic Uremic Syndrome (HUS) is a complication of Shigatoxin-producing Prospective study during 2017 – 2020 was performed; three groups of patients < 18 years were included: bloody diarrhea (BD), HUS requiring dialysis (HUSD) and HUS and no dialysis requirement (HUSND), all of them with presence of STX in stool. Blood samples were collected at diagnosis (t1) and at 7 days (t2). An immunoassay was used for detection of TNF-α, IL-1β, IL-6, and IL-8 . An immunoassay  was used for N-gal detectionForty-nine children were admitted; 22 (49%) were male. Median age was 24 (IQR 13-36) months. Fourteen patients with BD, 24 patients with HUS: 12 HUSD and 12 HUSND. Eleven healthy children (HC) were included. At diagnosis, higher IL-8 values were found in HUSD vs. BD (p=0.0004), HUSND (0.003) and HC (p= 0.0043). Higher TNF-α values were detected in HUSD vs. BD (P= 0.0001), HUSND (p= 0.0011) and HC (p= 0.0002). Higher TNF-α values were found in patients with BD vs HC (p= 0.0055). HUSD exhibited higher IL-6 values as compared with BD (p= 0.0145) and HC (0.0106). N-gal concentrations were higher in HUSD vs. BD (P= 0.0120), HUSND (P= 0.0166) and HC (p= 0.0049). A decrease in IL-8 was observed in HUSD between t1 and t2 (p=0.0357). Interleukin 8, IL-6, and N-gal may be considered as potential serological markers of renal damage and dialysis requirement. TNF-α concentrations are already increased in BD without HUS complicationsAll Authors: No reported disclosures.


In the published article, the reference for Immunotherapy is an emerging tool used in cancer treatment (1) was incorrectly written as Ruhlmann CH, Iversen TZ, Okera M, Muhic A, Kristensen G, Feyer P, et al. Multinational study exploring patients’ perceptions of side-effects induced by chemo-radiotherapy. Radiother Oncol (2015) 117(2):333–7. doi: 10.1016/j.radonc.2015.09.014. It should be Mohanty R, Chowdhury CR, Arega S, Sen P, Ganguly P, Ganguly N. CAR T cell therapy: A new era for cancer treatment (Review). Oncol Rep (2019) 42(6):2183-95. Epub 2019/10/04. doi: 10.3892/or.2019.7335.In the published article, the reference for The fourth-generation CARs added genes encoding cytokines [interleukin (IL)-12 and IL-15] to be released by the CARs to improve CAR T-cell survival in a TME (32) was incorrectly written as Haso W, Lee DW, Shah NN, Stetler-Stevenson M, Yuan CM, Pastan IH, et al. Anti-Cd22-chimeric antigen receptors targeting B-cell precursor acute lymphoblastic leukemia. Blood (2013) 121(7):1165–74. doi: 10.1182/blood-2012-06-438002. It should be Chmielewski M, Kopecky C, Hombach AA, Abken H. IL-12 release by engineered T cells expressing chimeric antigen receptors can effectively muster an antigen-independent macrophage response on tumor cells that have shut down tumor antigen expression. Cancer Res (2011) 71(17):5697-706. Epub 2011/07/12. doi: 10.1158/0008-5472.Can-11-0103.In the published article, the reference for The fifth-generation CARs build on the second-generation CARs by adding cytoplasmic structural domains from the IL-2 receptor beta chain and signal transducers and activators of transcription (STAT)3/5 binding pattern, triggering three signals including T-cell receptors , costimulatory factors , and cytokines  to improve the proliferation, survival, and antitumor activity of CAR T cells markedly (33) was incorrectly written as Chmielewski M, Kopecky C, Hombach AA, Abken H. IL-12 release by engineered T cells expressing chimeric antigen receptors can effectively muster an antigen-independent macrophage response on tumor cells that have shut down tumor antigen expression. Cancer Res (2011) 71(17):5697–706. doi: 10.1158/0008-5472.Can-11-0103. It should be Kagoya Y, Tanaka S, Guo T, Anczurowski M, Wang CH, Saso K, et al. A novel chimeric antigen receptor containing a JAK-STAT signaling domain mediates superior antitumor effects. Nat Med (2018) 24(3):352-9. Epub 2018/02/06. doi: 10.1038/nm.4478.In the published article, the reference for As conventional surgical treatment and antitumor drugs have limited effects, CAR T-cell therapy can provide targeted immunotherapy to patients with gastric cancer without developing drug resistance and effectively control the progression and metastasis of gastric cancer (62) was incorrectly written as Durães C, Almeida GM, Seruca R, Oliveira C, Carneiro F. Biomarkers for gastric cancer: Prognostic, predictive or targets of therapy? Virchows Arch (2014) 464(3):367–78. doi: 10.1007/s00428-013-1533-y. It should be Jiang H, Shi Z, Wang P, Wang C, Yang L, Du G, et al. Claudin18.2-specific chimeric antigen receptor engineered T cells for the treatment of gastric cancer. J Natl Cancer Inst (2019) 111(4):409-18. Epub 2018/09/12. doi: 10.1093/jnci/djy134.In the published article, the reference for Animal models are constructed by gene editing to mimic specific biological characteristics of human diseases to introduce target genes or delete and modify endogenous genes (83) was incorrectly written as Hu W, Lazar MA. Modelling metabolic diseases and drug response using stem cells and organoids. Nat Rev Endocrinol (2022) 1–16. doi: 10.1038/s41574-022-00733-z. It should be Platt RJ, Chen S, Zhou Y, Yim MJ, Swiech L, Kempton HR, et al. CRISPR-Cas9 knockin mice for genome editing and cancer modeling. Cell (2014) 159(2):440-55. Epub 2014/09/30. doi: 10.1016/j.cell.2014.09.014.The authors apologize for these errors and state that these do not change the scientific conclusions of the article in any way. The original article has been updated.


Frequent users of Emergency Departments (EDs) are a diverse group accounting for disproportionate EDs visits. Psychiatric patients are more likely to visit EDs . EDs utilisation by psychiatric patients increased by 4.4% during COVID-19 pandemic.to determine frequent users characteristics within an Ottawa University Hospital, and assess Covid19 impact on overutilization of EDs compared to other hospitals.Retrospective study of repeat visits characteristics, data extracted from EMR database. Repeat visits defined as no less than 30 days first visit to any EDs. Period of observation: March 1st, 2018 - February 28th, 2021 Results.64% EDS visits for MH, 35% for addictions. More men (57%), age groups: 16-34 y.o. (41%), 34-64 y.o. (51%), 65 +y.o. (8%).Top presenting reasons: suicidality, self-harm, depression (40.5%). Anxiety, situational crisis (16%), bizarre behavior (12%).Most prevalent diagnoses: schizophrenia (28.7%), stress and anxiety (25.2%), personality disorders (13.5%) and depressive episode (10.6%). Only 35.1% admitted after repeat ED visits, 35.1% came by ambulance. Increase during peak pandemic exceeding 20%. Clearly pandemic created more pressures for MH services needs.Schizophrenia and personality disorders made most prevalent diagnostic groups. Even when patients are in acute needs, they do not always require hospitalization. Investigating what MH conditions that got more stressed by the Covid19 pandemic will be of interest.No significant relationships.


Gossypium raimondii Ulbrich (short fibers) with cultivated diploid G. arboreum L and tetraploid G. hirsutum L. (long fibers); 2) G. hirsutum short fiber mutants, Ligon-lintless 1 (Li1) and 2 (Li2) with their near isogenic line (NIL), DP-5690 (long fibers). Chemical analyses showed that the short fibers commonly consisted of greater non-cellulosic components, including lignin and suberin, than the long fibers. Transcriptomic analyses also identified up-regulation of the genes related to suberin and lignin biosynthesis in the short fibers. Our results may provide insight on how high levels of suberin and lignin in cell walls can affect cotton fiber length. The approaches combining phenomic and transcriptomic analyses of multiple sets of cotton fibers sharing a common phenotype would facilitate identifying genes and common pathways that significantly influence cotton fiber properties.Fiber length is one of the major properties determining the quality and commercial value of cotton. To understand the mechanisms regulating fiber length, genetic variations of cotton species and mutants producing short fibers have been compared with cultivated cottons generating long and normal fibers. However, their phenomic variation other than fiber length has not been well characterized. Therefore, we compared physical and chemical properties of the short fibers with the long fibers. Fiber characteristics were compared in two sets: 1) wild diploid  Gossypium sp.) is the most economically important natural fiber in the world . Pe. PeArabiCW stage . Three Gi fibers .Li1 and Li2) was performed with total RNAs extracted from developing fibers at PCW stage (8–12 DPA) grown in greenhouse or cotton fields [Li1 and Li2 fibers were compared to their NIL, G. hirsutum DP-5690, using a 2-fold difference as a threshold. In the Li1 mutant fibers, 4,043 genes were up-regulated whereas 2,536 genes were down-regulated  was performed with the spectral region (1200–3000 cm-1) composed of IR peak bands of suberin, lignin, and cellulose (G. hirsutum Li2 < G. hirsutum Li1 < G. raimondii D5-31 < G. hirsutum TM-1 ≈ G. hirsutum DP-5690 ≈ G. arboreum A2-100. The three cultivated cottons of G. arboreum A2-100 (0.369), G. hirsutum DP-5690 (0.365), and G. hirsutum TM-1 (0.326) demonstrated similar positive PC1 scores with insignificant (p = 0.587) variations. In contrast, the other three short fiber cottons of G. hirsutum Li2 (-0.738), G. hirsutum Li1 (-0.279), and G. raimondii D5-31 (-0.043) had all negative scores that were significantly (p<0.0001) variable from the cultivated cotton species. Among the three short fiber cottons, the PC1 scores also showed significant (p<0.0001) variations. These results suggested that the chemical compositions of G. raimondii D5-31, G. hirsutum Li1, and G. hirsutum Li2 were different despite a sharing of suberin and lignin fractions in addition to cellulose.For an examination of the quantitative and statistical significances of the spectral features showing different chemical components among the fiber samples used in the 1ellulose . The anaellulose . The PC1G. hirsutum Li1  and Li2  fibers (G. hirsutum Li1 (880) and Li2 (376) mutants. To identify the commonly up-regulated orthologous genes between the diploid D5 genome and polyploid AD1 genome composed of AT and DT subgenomes, we compared the diploid G. raimondii 1,385 SEGs with non-redundant UGs in polyploid G. hirsutum Li1  and Li2  (T) and a peroxidase (Gorai.013G005800_AT) involved in lignin polymerization by oxidizing lignin monomers (monolignols) [T) required for suberin biosynthesis in Arabidopsis [Arabidopsis color mutant was also up-regulated [Of the UGs in ) fibers , there w  . Among t  . Consistlignols) , 53, 54 bidopsis  were foubidopsis  was alsoegulated . The othegulated , leucineegulated , FAD-binegulated , glutathegulated , PLAT/LHm Li1 3,1 and Li2 egulated , cytokinegulated , PSBP-doegulated , cyclic egulated , and NACegulated  were allG. raimondii and G. hirsutum Li1 and Li2 mutants were too short to be measured by HVI. Thus, we manually measured the maximum fiber lengths of the wet and relaxed cotton fibers from the chalezel end of cottonseeds [G. raimondii D5-6 (11.7 mm) and D5-31 (10.1 mm) as well as G. hirsutum Li1 (2.6 mm) and Li2 (8.1 mm) mutants were significantly shorter than those of cultivated diploid cotton, G. arboreum SXY1 (28.0 mm) and A2-100 (30.1 mm) as well as cultivated polyploid cotton, G. hirsutum TM-1 (41.7 mm), SG-747 (40.1 mm) and DP-5690 (38.7 mm) as shown in Figs G. hirsutum fibers. Lint fibers differentiate from ovule epidermis on the day of anthesis and they grow approximately 25~35 mm based on HVI measurements. In contrast, linter or fuzz differentiate from the ovule epidermis around 5 to 10 DPA and they do not grow longer than 15 mm [G. raimondii was often described as a lintless, non-fibered, or fiberless species [Li1 and Li2 mutants also contain “lintless” [G. raimondii [G. hirsutum Li1 mutant [G. hirsutum Li2 mutant [G. raimondii and G. hirsutum Li1 and Li2 mutants can be classified as lint fibers according to the definition described by Lang [Physical properties of cultivated cotton fibers are generally assessed by a High Volume Instrument (HVI) which is defined by the International Cotton Advisory Committee as a standardized instrument for cotton fiber quality measurements . Cotton tonseeds . The maxan 15 mm . Wild di species , 69, 70.intless” , 18. How1 mutant , and G. 2 mutant  all diff by Lang .G. raimondii and two G. hirsutum mutants produced fibers containing color pigments composed of lignin and suberin  and G. hirsutum Li1 and Li2 mutants (85.1~86.9%) fibers were lower than cultivated G. arboreum (95.6~100%) and G. hirsutum (95.8~98.0%) fibers  and G. hirsutum Li1 and Li2 (13.1~14.9%) fibers were substantially greater than the cultivated fibers (0~4.4%). These results were consistent with the previous reports showing different cellulose contents between green and white upland cotton [G. raimondii and cultivated G. arboreum [Chemical analyses using cellulose assay, ATR FT-IR spectroscopy, and mass spectrometry consistently showed suberin and lignin components in the three short fibers. Average cellulose contents of ers Figs  and 3B. d cotton  and funcarboreum .G. raimondii and G. hirsutum Li1 and Li2 mutants demonstrated the signature IR spectral peaks of suberin and lignin  than those of the other cultivated cotton fibers (0~ 0.6%). Recent studies suggest that lignin may play an important role in cotton fiber quality [The three short cottons of nin Figs  and 5B.  suberin . In natun fibers , 74, andn fibers . Suberinn fibers . In contn fibers . Generaln fibers . Our mas quality , 79.G. hirsutum Li1 (1623 cm-1), G. hirsutum Li2 (1610 cm-1), and G. raimondii D5 (1635 cm-1)  Figs  and 5B. -1) Figs .G. raimondii fibers, we used the RNA-seq data performed with the RNAs extracted from developing fibers of G. raimondii, G. arboreum, and G. hirsutum (Li1 and Li2) and their NIL DP-5690 were only performed with total RNAs extracted from developing fibers at PCW stage [G. raimondii fibers [G. raimondii D5 reference genome for analyzing transcriptomic profiles of the two sets because the G. raimondii genome sequence shows high homology (>96%) with the coding sequences of G. hirsutum AT and DT subgenomes. Thus, the D5 reference genome sequence has been successfully used for characterizing the Li1 and Li2 genomes by several groups [st set identified that genes involved in suberin and lignin biosynthesis were specifically expressed in G. raimondii fibers Click here for additional data file.S1 Table(XLSX)Click here for additional data file.S2 Table(DOCX)Click here for additional data file.S3 Table(XLSX)Click here for additional data file.S4 Table(XLSX)Click here for additional data file.S5 Table(XLSX)Click here for additional data file.


Dear Editor,3. The intrinsic nature of MLKL and how it induces plasma membrane permeabilization, forming a huge pore or a channel, remain an interesting conundrum for a long time5. Our previous study demonstrated that MLKL forms cation channels and its channel activity is a primary effector of necroptosis6. In the field of MLKL, a major unsettled issue is, if MLKL does serve as channels, how it ignites the necrotic or non-necrotic pathogenic progresses5. Phosphatidylinositol 4,5-bisphosphate P2) is a necessary cofactor of various ion channels7. The physical interaction between MLKL and phosphatidylinositol phosphates (PIPs), including PIP2, facilitates MLKL-mediated liposome leakage and the necrotic membrane disruption10. These studies have been directed toward understanding whether PIP2 is a direct modulator for the MLKL channel activity. The potential effects of PIP2 on MLKL channels were thus evaluated electrophysiologically and the subsequent pathogenic influences were explored.Mixed lineage kinase domain-like protein (MLKL) emerged as executioner of necroptosisNT) is sufficient to induce oligomerization and trigger cell death9. Consistent with our previous study, MLKLNT protein exhibited channel activity, translocated onto plasma membrane, and caused cell death, similarly to full-length protein (MLKLFL) P2  was enhanced to 27.4%  and phosphatidylinositol 3,5-bisphosphate P2), phosphatidylinositol 4-phosphate (PI(4)P), 1,4,5-trisphosphate inositol (IP3) and diacylglycerol (DAG), phosphatidylinositol 3,4,5-trisphosphate (PIP3)11. Among these lipids, PI(4)P is the only one that increases MLKL channel open probability P2. An increased concentration of PIP2 induced more frequent and larger currents of MLKL channel P2 levels in live cells. Stimulation of the M1 muscarinic receptor can activate PLC-β which hydrolyzes PIP2. PLC-β-PH-GFP (PLC-PH) construct, a PIP2 reporter, is used to monitor the PIP2 distribution on plasma membrane12. Here, MLKL, M1 receptor, and PLC-PH were co-transfected into HEK293 cells. With 5 μM oxotremorine M (Oxo-M) treatment, the PLC-PH probe dissociated from the plasma membrane to cytoplasm, showing PIP2 hydrolyzation  in the N-terminal region of MLKL (MLKL)P2 Fig. . The ove.4% Fig. . These s.4% Fig. . Influenity Fig. . Collectnel Fig. . To furtion Fig. . Meanwhiion Fig. . These d2-induced augmentation of MLKL channel activity is correlated with cell death was examined. To elevate PIP2 concentrations in the plasma membrane, PIP5K, one of the key PIP2 biosynthetic enzymes, was transfected into L929 cells P. 1 Fig. . In addi. 1 Fig. . Similar2-sensitive ion channels, PIP2 binds with positive amino acid via “electrostatic interaction”13. We mutated the positive amino acids of MLKLNT to alanine and tested the channel activity  P2 sensitivity or abrogate the capability of killing cells after overexpression of PIP5K, suggesting that K22 and R34 may interact with PIP2 independently P. 2 Fig. . Notably4%) Fig. . Whetherls9 Fig. . Consisttly Fig. .14. Thus, we further explored the potential function of PIP2-enhanced MLKL channel activity in an inflammation model12  through String database (https://string-db.org), and further revealed the close relationship between PIP2, MLKL and LPS-induced inflammation -like receptor protein 3 (NLRP3) in a cell-intrinsic manner before cell lysis, but its working model has not yet been clarifiedins Fig. . To inveion Fig. .+ efflux in triggering inflammation has been proposed15. We therefore hypothesized that K+ efflux during inflammation is mediated by MLKL channels. A real-time, sensitive FluxOR™ assay was performed to monitor the K+ efflux during the LPS-induced inflammation. We found that the K+ efflux gradually increased in a time-dependent manner  treated with LPS, Smac-mimetic compound (C) and a caspase inhibitor Q-VD-OPh (Q)14. Both the MLKL-mediated cytokine secretion and the LPS-induced K+ efflux were abrogated in MLKL-knockout (MLKL–/–) cells  of LPS significantly stimulated the secretion of inflammatory cytokines, but did not induce cell death, while the expression level of MLKL and PIP5K increased Fig. . Accordiner Fig. . The conlls Fig. . Whetherels Fig. . Since e2 in a dose-dependent manner for the first time Pime Fig. . PIuli Fig. . WhetherSupplementary informationSupplementary Tabel S1