One of the most damaging aspects, both for people’s well-being and for close relationships, is conflict. Beyond different stressors, the emotions evoked, their regulation and an appropriate conflict resolution strategy will reduce negative consequences. Emotional Intelligence facilitates social relationships, but little applied research has been done on the relationship with couple conflict and emotional flooding, particularly from the perspective of women. Therefore, the present study analyzes the role of Perceived Emotional Intelligence (PEI) and the mediating effect of Positive Conflict Resolution strategies (PCR) in couples’ conflicts from women’s perspective, examining its effect on Emotional Flooding (EF) and Satisfaction.Through a cross-sectional design, the relationships between variables were analyzed using group comparisons and means of a structural equation model (SEM) in a sample of 692 women.Significant differences were found between the groups by age, length of relationship, and motherhood. The SEM revealed a good fit. PEI predicted 71.8% of the variance in EF and 35% in Satisfaction through PCR and Conflict. As a re1.1.Emotions play a crucial role in relationship dynamics, with their elicitation and management being essential factors that can significantly influence conflict outcomes . EmotionAs shown, EI has a significant impact on individuals’ lives and reactions , and an 1.2.High emotional arousal in interpersonal conflict situations can lead to “flooding,” a state of overstimulation, overwhelm and cognitive disorganization . This teAs 1.3.Couple relationships differ from other relationships given the presence of certain unique processes and expectations such as fidelity and emotional and romantic exclusivity . One of On the basis of the evidence described above, we conducted the present research in an all-female sample with a 2-fold aim. On the one hand, we sought to extend the literature on women’s relationships by exploring group differences by age and relationship length on the endogenous variables of PCR, Conflict, EF, and Satisfaction. We also wanted to see if having children had any impact on Conflict and Satisfaction. On the other hand, we want to analyze the relationship between PEI, PCR, Conflict, EF, and Satisfaction by building an explanatory model using structural equations in this sample of women. To this end, the following hypotheses were formulated:H1: PEI is direct and positively related to PCR.H2: PEI is direct and negatively related to Conflict.H3: PCR is direct and negatively related to Conflict.H4: Conflict is direct and positively related to EF.H5: Conflict is direct and negatively related to Satisfaction.H6: EF is direct and negatively related to Satisfaction.2.2.1.This study followed a cross-sectional design, with participants being recruited through an online questionnaire and relying on non-probabilistic, snowball sampling that was distributed across distinct social media directed at Spanish residents. The hypothetical model based on the theory, together with the proposed hypotheses, is shown in 2.2.Participation in this study was voluntary and no compensation or payment was given to the participants. After an initial literature review and selection of instruments, an online questionnaire was created in Google Docs and the link was distributed through several social networks such a Twitter, Facebook. and Instagram. Participants were asked to share the link with friends and acquaintances. The study objectives were explained along with the potential benefits of participation. Informed consent was requested from the participants to process their data. A total of 825 responses were received. As the purpose of the study was to analyze the functioning of the variables from a gender perspective, only responses from female participants who were currently or previously in a relationship with a heterosexual partner were selected. The sample size exceeded the minimum required considering the number of latent constructs and items  This stuSD = 7.99). Of these, 64.6% were involved in a couple relationship, 21.7% were married, 11.0% single, 2.3% divorced, and 0.4% widowed. Of the selected sample, 28.0% had children. To make comparisons between groups, the sample was divided into three age groups and four groups according to the length of the relationship. Age groups were created following the recommendations of The final sample consisted of 692 women with a mean age of 31.76 years  by ad hoc questionnaire measuring conflict in couples were used.A sociodemographic data questionnaire was used in this study. It was an 2.3.1.strongly disagree) to 5 (strongly agree). Studies that analyze this factorial structure have revealed the existence of three dimensions or factors: (1) Emotional Attention: Consisting of items that comprise the capacity to pay attention to feelings and emotions ; (2) Emotional Clarity: Consisting of items indicating the ability to understand emotional states and to discriminate between different emotions and feelings ; and (3) Emotional Repair: Consists of items determining the ability to appropriately regulate one’s emotional states . The results are classified into three groups, according to the authors’ recommendations: low, good, and excellent skills in factors 2 and 3, and low, good, and excessive skills in factor 1. Prior studies have analyzed the scale’s internal consistency and validity .Perceived emotional intelligence was measured using the validated Spanish version by validity . Its rel2.3.2.almost never) to 5  measuring the degree to which the individual used positive problem-solving strategies  in response to conflicts, with questions about the frequency of situations such as “sitting down and discussing our differences constructively.” Prior studies have analyzed the internal consistency and validity of the inventory (The PCR used by the couple was measured using the positive problem-solving dimension of Conflict Resolution Style Inventory (CRSI) according to the participants’ perspective in an adapted version that was validated in Spanish . It consnventory . Its rel2.3.3.completely disagree) to 5 (completely agree). It is based on four factors, according to the original proposal made by my partner’s negative attitudes overwhelm me”; the Unjustified Anger factor was measured using items such as “I get overly upset when I argue with my partner,” the Motivation to Run Away factor was measured using items such as “when we argue I want to escape and have nothing more to do with the relationship” and the Emotional Self-Regulation factor was measured using items such as “I find it difficult to control my anger when arguing with my partner.” The reliability of the scale in the sample was high (α = 0.92) as was that of the factors .Emotional Flooding was measured using the Scale of Emotional Flooding in Couple Relationships (EDEP) by 2.3.4.completely dissatisfied) to 7 (completely satisfied). Satisfaction is measured using items such as “How satisfied are you with your current relationship or marriage?.” The reliability of the scale in the sample was high (α = 0.89).Satisfaction was measured using the validated Spanish version by 2.3.5.ad hoc questionnaire based on the multidimensional model of emotion and How often do you and your partner argue? (b) When my partner and I disagree, we discuss it calmly, and (c) Arguing with my partner is very intense. Responses were measured on a five-point Likert-type scale ranging from 1 (completely disagree) to 5 (completely agree). The reliability of the scale in the sample was acceptable (α = 0.70).An 2.4.post hoc Dunn’s adjustment test was performed to examine significant differences between groups. Effect sizes were calculated using r and Cohen’s d.The SPSS 18.0, AMOS 23.0, and JASP 0.17.1 statistical packages were used for descriptive analysis, correlations, factorial analyses, and between-groups comparisons. Data were preliminarily screened for outliers and errors (not detected). For correlations, Spearman’s rho coefficient was used. The normality of the variables was verified with the Kolmogorov–Smirnov test and homoscedasticity was examined using Levene’s test for equality of variance. Due to the lack of assumptions required for parametric tests such as normality and symmetry, Kruskal-Wallis H and Mann–Whitney U tests were used to compare groups. For all of these tests, an alpha level of 0.05 was used. To correct the possible error resulting from the 5% threshold, the Bonferroni correction was used. The Confirmatory factorial analyses (CFA) were used to assess the psychometric properties of the instruments, as well as their factorial structure. Varimax rotation was used for the extraction of the main components. Estimation, identification, assessment, and re-specification of the structural equation model (SEM) were performed in AMOS. A variance–covariance matrix was used as well as maximum likelihood estimation. When estimating the parameters that define the factors, as well as the relations between variables, recommendations of For path analysis, SEM was used. Direct and indirect effects were calculated using a bootstrap technique to determine significance. A bootstrap sample of 5,000 was used. Significance was assessed using the bias-corrected percentile method with a 95% confidence interval. A multicollinearity test was performed with negative results. The goodness of fit parameters were estimated using a maximum likelihood estimation model. Given the lack of consensus regarding the concept of fit, various indices were considered simultaneously  to analy3.3.1.3.2.M = 6.740, SD = 1.819) and Middle-Aged adults  groups H (2) = 11.914, p < 0.05, with higher conflict scores in the MA group, with a moderate effect size d = 0.350, 95% CI = 0.071–0.630. By relationship length, significant differences were found between <3  and + 11  groups as well as between 3–7  and + 11 groups H = 15.32 (3), p < 0.001, with moderate effects sizes d = 0.415, 95% CI = 0.125–0.704 and d = 0.356, 95% CI = 0.063–0.649, respectively.Significant differences in Conflict by age were found  between M = 15.740, SD = 2.656) and Older Adults  groups H (2) = 7.264, p < 0.05, with moderate effect size d = 0.443, 95% CI = 0.030–0.916. By relationship length, significant differences were found H (3) = 20.080, p < 0.001 between <3  and + 11  groups, 3–7  and + 11 groups, as well as 7–11  and + 11 groups, with moderate effects sizes d = 0.420, 95% CI = 0.131–0.710, d = 0.484, 95% CI = 0.190–0.778 and d = 0.300, 95% CI = 0.027–0.627, respectively.In terms of PCR, significant age differences were found  between p < 0.001 between YA  and MA  groups as well as YA and OA  groups, with a moderate effect size d = 0.293, 95% CI = 0.013–0.572 and medium effect size d = 0.522, 95% CI = 0.048–0.995, respectively. By relationship length, significant differences were found H (3) = 16.407, p < 0.001 between <3  and + 11  groups, 3–7  and + 11 groups as well as 7–11  and + 11 groups, with moderate effect sizes d = 0.382, 95% CI = 0.093–0.672, d = 0.221, 95% CI = 0.071–0.514 and d = 0.250, 95% CI = 0.077–0.577, respectively.Regarding EF  significM = 12.967, SD = 1.902) and MA  groups H (2) = 18.163, p < 0.001 with moderate effect size d = 0.357, 95% CI = 0.077–0.636 as well as between YA and OA  with a medium effect size d = 0.697, 95% CI = 0.222–1.171. By relationship length, significant differences were found H (3) = 20.230, p < 0.001 between <3  and + 11  groups with moderate effect size d = 0.392, 95% CI = 0.103–0.682, 3–7  and + 11 groups with moderate effect size d = 0.408, 95% CI = 0.115–0.702, as well as 7–11  and + 11 groups, with moderate effect size d = 0.462, 95% CI = 0.133–0.790.Satisfaction also was significant when comparing groups . By age,Mdn = 7, n = 194) U = 37137.000, z = 4.795, p < 0.001, r = 0.18 and higher scores on Satisfaction in the group without children  U = 38926.500, z = 4.134, p < 0.001, r = 0.16.Finally, a significant difference was found when comparing the groups according to whether they had children or not, with higher levels of conflict found in the group of couples with children  b = 0.000, p = 0.992 were found to be insignificant and therefore dropped from the model, so H2 and H6 were not supported. The definitive, simplified SEM along with the standardized structural coefficients is shown in 2 = 3.373, p < 0.001; RMSEA = 0.059, 95% CI: 0.052, 0.065; NFI = 0.923; TLI = 0.931; CFI = 0.944; GFI = 0.941 and AGFI = 0.918. The direct effect of PEI on PCR was found to be positive and significant b = 0.032, p < 0.001, supporting H1. The direct effect of PCR on Conflict was found to be negative and significant b = −0.711, p < 0.001, supporting H3. The direct effect of Conflict on EF was found to be positive and significant b = 4.465, p < 0.001, supporting H4. Finally, the direct effect of Conflict on Satisfaction was found to be negative and significant b = −0.791, p < 0.001, supporting H5.An exploratory factor analysis was conducted to determine the behavior of the conflict questionnaire items and their construct validity. A confirmatory factor analysis was conducted with all other variables included in the model and all the scales retained the original item and factor structure. After the first analysis, the relationship between PEI and Conflict (H2) In summary, all hypotheses except H2 and H6 were supported and a significant indirect effect was found between PEI and Conflict, EF and Satisfaction through the mediators , explain4.The aim of the present study was to better understand the relationship between PEI and Conflict in a sample of women, analyzing the role of PCR, EF and Satisfaction. To this end, the relationships between these domains were examined using a full structural equation model. Differences between groups by having children, age, and relationship length were also examined, with the aim of extending the evidence in the women’s literature on couples, and significant results were found. Our results showed an escalation in levels of conflict as a function of age and relationship length, with women aged 40–49 (middle-aged) having higher levels of conflict than younger women. Similarly, women who had been in a relationship for more than 11 years had higher levels of conflict with their partner than women who had been in a relationship for less than 3 years or between 3 and 7 years. This finding is consistent with previous research showing that older married women tend to report more anger than middle-aged adults in an experiment involving disagreements with their husbands . AlthougThe results of the SEM confirmed the role of PCR and Conflict as mediators explaining the relationship between PEI, EF, and Satisfaction in our sample. The need to apply appropriate conflict resolution strategies to positively resolve couple relationship problems has been clearly demonstrated in numerous studies , which i5.Couple relationships are a complex environment in which different episodes, emotions and situations occur that affect both the couple’s well-being and several individual variables. To our knowledge, this is the first study to examine the importance of Perceived Emotional Intelligence and Positive Conflict Resolution strategies in situations of partner conflict, explaining Emotional Flooding and satisfaction as outcomes from a female perspective. The findings highlight the need for adequate Perceived Emotional Intelligence and Positive Conflict Resolution strategies to prevent partner conflict, reduce the occurrence of Emotional Flooding, and increase relationship satisfaction. These results would justify the need to work on emotional competences together with Positive Conflict Resolution strategies in any couple context in order to improve interpersonal relationships, whether at the clinical, intervention, or prevention program level. In addition, we found relevant evidence to add to the women’s literature when analyzing groups. Conflict increased as a function of age and relationship length, as did Emotional Flooding, while Positive Conflict Resolution strategies and Satisfaction decreased.5.1.The study’s primary participants are individuals who identify as heterosexual, are in monogamous, committed relationships—including marriage—and have been in relationships for a significant length of time. Although this selection criterion was motivated by the fact that most previous research on intimate relationships has focused on the experiences of heterosexual couples, caution should be exercised in generalizing the results. Additionally, the vast majority of the sample had higher education levels and the age sample was unbalanced, which may limit the generalisability of the findings to other social groups. To provide a more comprehensive understanding of the implications of the relationships found in the model, a longitudinal study could be conducted to analyze the functioning of couples over time and observe changes in the variables of interest. Future research could also explore the role as potential risk or protective factors in intimate partner violence.https://osf.io/ng5ku/?view_only=f450eb6faad24e7ebe4db2b7a9e0ac4c.The datasets presented in this study can be found in online repositories. The names of the repository/repositories and accession number(s) can be found at: The studies involving human participants were reviewed and approved by Research Ethics and Integrity Committee of Miguel Hernández University, Spain (DPS.AGL.01.21). The patients/participants provided their written informed consent to participate in this study.AG and MB-S designed the study and oversaw all aspects of study implementation. AG and MB-S collected the data and DP managed the database. AG and DP performed the statistical analyses and edited the final draft. MB-S wrote the first draft of the manuscript. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Anchistea, Claytosmunda, Dryopteris, Osmunda, Osmundastrum, Equisetum) and bryophytes . Our findings also suggest that honey bees may occasionally act as vectors for the dispersal of aquatic phototrophs, specifically Coccomyxa and Protosiphon, species of green algae. Our work has shed light on the broad resource‐access patterns that guide plant‐pollinator interactions and suggests that bees could act as vectors of gene flow, and potentially even agents of selection, across Plantae.Honey bees are efficient pollinators of flowering plants, aiding in the plant reproductive cycle and acting as vehicles for evolutionary processes. Their role as agents of selection and drivers of gene flow is instrumental to the structure of plant populations, but historically, our understanding of their influence has been limited to predominantly insect‐dispersed flowering species. Recent metagenetic work has provided evidence that honey bees also forage on pollen from anemophilous species, suggesting that their role as vectors for transmission of plant genetic material is not confined to groups designated as entomophilous, and leading us to ask: could honey bees act as dispersal agents for non‐flowering plant taxa? Using an extensive pollen metabarcoding dataset from Canada, we discovered that honey bees may serve as dispersal agents for an array of sporophytes  and bryophytes . Our findings also suggest that honey bees may occasionally act as vectors for dispersal of aquatic phototrophs, specifically Coccomyxa and Protosiphon, species of green algae. These findings, though preliminary, open up a new field of research.We used pollen metabarcoding to investigate if honey bees interact with and disperse cells from non‐flowering plants. We discovered that honey bees may serve as dispersal agents for an array of sporophytes ( In partnership with local beekeepers and the Ontario Beekeepers Association, we placed 50 honey bee colonies at 24 sites in Southern Ontario Figure . We harv2.2We utilized our previously validated pollen metabarcoding protocol . We pipetted 11 μL of sterile H2O, 12.5 μL of 2× Taq Pol Mix (New England Biolabs), 0.5 μL of each relevant forward and reverse primer (1 μM), and 0.5 μL of sample DNA (~36.2 ng) into each well. We then ran a PCR cycle  using the following program specifications: initial denaturation , denaturation , primer annealing , extension , final extension . PCR1 product was used as the template for PCR2; the same master mix described above was combined with 0.5 μL of PCR1 product. Program specifications were maintained between PCR1 and PCR2, with one change to the primer annealing temperature (56°C). Following PCR2, we prepared samples for Illumina Sequencing by performing a third PCR program that tagged each sample with a unique combination of forward and reverse indexes; PCR3 program specifications follow that described above, with a primer annealing temperature of 60°C. We then normalized the resulting PCR3 product using a SequalPrep Normalization Kit (Invitrogen) and shipped the normalized samples on dry ice for Illumina Sequencing at Genome Quebec.We performed two PCR programs, the first of which amplified the region of interest PCR1), and the second of which extended the target sequence with a read priming sequence (PCR2), for each of the two primers  employed as part of our metabarcoding program , and R (v. 4.2.1), using the 3incertae sedis), Protosiphonales, Osmundales, Polypodiales, Equisetales, Sphagnales, Orthotrichales, and Funariales. This indicated the presence of three novel dietary constituents – phytoplankton, sporophytes, and bryophytes (Table rbcL).After filtering out all sequence hits associated with angiosperm and gymnosperm species and blasting all remaining sequences through the NCBI database to confirm sequence identification, we found 37 ASVs (amplicon sequence variants) associated with plant taxa of the orders Coccomyxaceae . ASVs associated with fern species were much more common, encompassing five different genera across 17 ASVs. The most abundant fern genus, Osmundastrum, had an average sequence match of 99.5% (±0.5). The second most abundant fern genus, Osmunda, had an average sequence match of 96% (±4.0); the remaining fern genera  were all associated with singular ASVs and an average sequence match of 99.6% (±0.1). The three remaining ASVs were linked to Coccomyxa and Protosiphon, genera of green algae, with an average sequence match of 92.7% (±2.6).Four genera of bryophytes were identified via our bioinformatics pipeline, with an average sequence match of 98.9% (±1.3); Osmundastrum, Dryopteris, Osmunda, Anchistea, Claytosmunda) and horsetails (Equisetum), but not any species of moss nor algae , but with lower sporophyte diversity than spring samples , including multiple species of ferns and mosses Table . Time poae Table . Time poes Table . Spring 4rbcL). Gous et al. ; data curation (lead); formal analysis (lead); investigation (lead); methodology ; writing – original draft (lead); writing – review and editing . Laura R. Newburn: Methodology ; writing – review and editing . Rodney T. Richardson: Methodology ; resources ; software ; writing – review and editing . Mateus Pepinelli: Investigation ; writing – review and editing . Ida M. Conflitti: Funding acquisition ; project administration (lead); resources ; writing – review and editing . Mashaba Moubony: Investigation ; writing – review and editing . Daniel Borges: Investigation ; writing – review and editing . M. Marta Guarna: Funding acquisition ; investigation ; resources ; writing – review and editing . Ernesto Guzman‐Novoa: Funding acquisition ; investigation ; resources ; writing – review and editing . Leonard J. Foster: Funding acquisition ; investigation ; resources ; writing – review and editing . Amro Zayed: Conceptualization (lead); funding acquisition (lead); investigation (lead); resources (lead); supervision (lead); writing – review and editing .The authors declare no competing interests.Appendix S1:Click here for additional data file.Video S1:Click here for additional data file.


EGFR) mutations have not been fully studied.The efficacy and safety of osimertinib combined with bevacizumab in non‐small cell lung cancer (NSCLC) patients with brain metastasis harboring epidermal growth factor receptor , and the secondary endpoints were the median overall survival (mOS), response rates, and toxicities. This study has been registered in ClinicalTrials.gov (NCT05104281) and is ongoing.Treatment‐naïve NSCLC patients with brain metastasis harboring EGFR 19 del and 35 harbored EGFR L858R mutation. The objective response rate (ORR) was 75.0% and the disease control rate (DCR) was 96.2%; the mPFS was 17.0 months (95% CI: 11.46–22.54), while the mOS was not reached. The mPFS was 20.0 months (95% CI: 14.56–25.44) and was 17.0 months (95% CI: 13.28–20.72) for patients harboring EGFR 19 del and EGFR L858R mutation (p = 0.844), respectively. The intracranial ORR was 82.7%, and the intracranial mPFS was 22.0 months (95% CI: 2.92–41.08).The main adverse events were mild‐to‐moderate hand‐foot syndrome, diarrhea, hypertension, and proteinuria. Three patients developed grade III proteinuria, while five patients developed grade III hypertension; they permanently discontinued bevacizumab treatment.A total of 52 Chinese patients were enrolled, of whom 17 harbored EGFR‐mutated NSCLC patients with brain metastasis, and the side effects are tolerable.Osimertinib combined with bevacizumab shows promising results in  EGFR‐activating mutations were treated with osimertinib and bevacizumab. A total of 52 Chinese patients were enrolled, of whom 17 harbored EGFR 19 del and 35 harbored EGFR L858R mutation. The objective response rate (ORR) was 75.0% and the disease control rate (DCR) was 96.2%; the mPFS was 17.0 months (95% CI: 11.46–22.54), while the mOS was not reached. The mPFS was 20.0 months (95% CI: 14.56–25.44) and was 17.0 months (95% CI: 13.28–20.72) for patients harboring EGFR 19 del and EGFR L858R mutation (p = 0.844), respectively. The intracranial ORR was 82.7%, and the intracranial mPFS was 22.0 months (95% CI: 2.92–41.08). Osimertinib combined with bevacizumab shows promising results in EGFR‐mutated NSCLC patients with brain metastasis, and the side effects are tolerable.Treatment‐naïve NSCLC patients with brain metastasis harboring  EGFR mutations, resistance would eventually occur with progression‐free survival (PFS) durations of 9.5–13.1 months for those treated with first‐ or second‐generation EGFR‐TKIs and 18.9 months for those treated with osimertinib.EGFR somatic mutations, laborious effects have been made continuously in combined models in recent years.EGFR‐activating mutations, but the OS was not significantly different.Lung cancer is the most common cancer type and the leading cause of cancer‐related deaths globally; non‐small cell lung cancer (NSCLC) accounts for 85%–90% of lung cancer cases.EGFR‐mutant lung cancers.EGFR or EGFR T790M mutations.EGFR gene mutation‐positive NSCLC are prone to developing brain metastases, with the frequency ranging from 44% to 63%.EGFR mutation patients with brain metastasis remains controversial;EGFR‐activating mutations.Osimertinib, a third‐generation EGFR‐TKI, demonstrated superior PFS and OS compared with erlotinib or gefitinib as the initial treatment for patients with EGFR exon 19 del or L858R mutations confirmed via next‐generation sequencing (NGS) of biopsied tissues and/or blood. Computed tomography (CT)‐guided core needle biopsy was performed, DNA was extracted from 15 × 5 μm sliced sections of formalin‐fixed, paraffin‐embedded tumor tissue, and the tumor area was evaluated by a pathologist. For adequate sequences and mutation detection, at least 20% of the tumor area on each slice was set as the minimum. A total of 10 mL of blood were drawn and centrifuged for sequencing control and germline gene mutation tests. NGS was performed using HiSeq3000/HiSeq4000 Illumina techniques. Approximately 4278 exons of 288 common genes; intron, promoter, and fusion of 38 genes; coding area of 728 genes were tested for somatic mutations. A total of 11 germline mutations were identified. The ultra‐deep coverages of genes of interest were 1000× for tumor tissue and 10 000× for serum.This is an ongoing, two‐center study. Patients enrolled in the study were histologically confirmed as having metastatic nonsquamous NSCLC with brain metastasis and were treatment naïve, with Participants were required to have at least one lesion in the lung and brain, which could accurately be measured in at least one dimension (longest diameter to be recorded) of ≥10 mm with spiral CT or magnetic resonance imaging (MRI). Patients with oligometastases in the brain and regional lung disease, which were suitable for surgical resection, were excluded. Meanwhile, participants aged ≥18 years, who had adequate organ and bone marrow function and had an Eastern Cooperative Oncology Group (ECOG) performance status score ≤2, were included in the study.Bevacizumab  15 mg/kg was administered intravenously on day 1 and repeated every 21 days; osimertinib  80 mg was administered orally once a day. Treatment was continued until disease progression, intolerable toxicity, or death occurred. Reduction of bevacizumab dose to 7.5 mg/kg or osimertinib dose to 80 mg every other day was permitted.Brain irradiation with whole‐brain radiotherapy (WBRT) or stereotactic radiosurgery (SRS) was indicated when patients had neurological symptoms or when it was required by a physician. The primary endpoint was PFS, and the secondary endpoints were OS, response rate, and toxicities.The primary endpoint of the study was PFS, while the secondary endpoints were OS, response rate, and toxicities. Responders were defined as those who had complete or partial response. PFS was measured from the first day of treatment until the first objective sign of disease progression or patient death, while OS was measured from the day of treatment to the day of patient death. Tumor responses, which were evaluated based on the Response Evaluation Criteria in Solid Tumor version 1.1 (RECIST version 1.1),Tumor measurements using chest CT or MRI were performed at baseline and every 8–12 weeks thereafter. Patient compliance, treatment safety, and side effects were assessed every 2–3 weeks, every check‐up.ClinicalTrials.gov (NCT05104281) and is ongoing.Based on the Cox proportional hazards model and taking into account the influence of sex , Eastern Cooperative Oncology Group (ECOG) performance status score, hazard ratio, and 95% confidence interval (CI) were calculated in the full analysis population. The PFS and OS curves were analyzed using SigmaPlot 14.0 (Systat Software Inc.). The Kaplan–Meier log‐rank test was used in the intention‐to‐treat (ITT) population. The Kaplan–Meier method was used for survival analyses, and the log‐rank test was used to test for significance. The response rate and treatment‐related adverse events were assessed using the Fisher's exact test. All enrolled patients who received at least one dose of the study treatment were included in the safety analysis. Periodic safety monitoring and interim efficacy assessments were performed by an independent data monitoring committee. This trial has been registered in The protocol and all modifications were approved by the Ethics Committee of the Affiliated Qingdao Central Hospital of Qingdao University on October 18, 2018  and were performed in compliance with the provisions of Good Clinical Practice guidelines, the Declaration of Helsinki, and local laws. Informed consent was obtained from all patients prior to enrolment.EGFR 19 del and L858R mutations were observed in 32.7% and 67.3% of the patients, respectively. All patients were newly diagnosed and treatment naïve. The demographic characteristics of patients are presented in Table A total of 71 patients were screened and 52 patients were subsequently enrolled in the study, of whom 38.5% (20/52) were male, and 61.5% (32/52) were female. The median age was 64.5 years, and 51.9% (27/52) of the patients were aged <65 years old. Approximately 90.4% (47/52) of the patients had adenocarcinomas, and 65.4% had an ECOG performance status score of 2. The objective response rate (ORR)  was 76.9% 40/52), which was assessed by the study investigators according to RECIST version 1.1, with a stable disease rate of 19.2% (10/52) and a disease control rate (DCR) of 96.2% (50/52). The median depth‐of‐response rate of the tumor (the nadir of tumor response) was 50.0%  Figure , the medEGFR 19 del and 17.0 months (95% CI: 13.28–20.72) in patients harboring EGFR L858R mutation (p = 0.844)  in patients harboring .56–25.44The side effects were mild‐to‐moderate hand‐foot syndrome, diarrhea, hypertension, and proteinuria. Three patients developed grade 3 proteinuria, while five patients developed grade 3 hypertension, for which bevacizumab treatment was permanently discontinued. Hand‐foot syndrome occurred in 80.8% (42/52) of the patients, which was the most common adverse effect, but most patients had grade I or II adverse effects. Five patients acquired paronychia, all of whom showed improvement after dose reduction and lotion embrocation (Table EGFR mutations are considered the most robust predictive biomarkers for clinical and radiographic responses to EGFR‐TKIs in clinical practice.EGFR‐activating mutations compared with gefitinib or erlotinib alone; the OS was not significantly changed.EGFR‐activating mutations, or in patients resistant to gefitinib or erlotinib who harbored the EGFR‐T790M mutation.EGFR mutations treated with EGFR‐TKIs showed disease progression in the central nervous system, such as brain metastases or leptomeningeal metastases.EGFR mutations. It had a high response rate and durable extracranial PFS and IC‐PFS. However, the present study had some limitations. This was a one‐arm study with fewer participants, which can potentially cause selection bias. Compared with other studies, the present study had a relatively short follow‐up period, which can affect the statistical power. The predictive value of osimertinib plus bevacizumab in the treatment of patients with NSCLC with brain metastasis harboring EGFR mutations should be further explored in a randomized setting.To the best of our knowledge, this study is the first to report the efficacy of osimertinib in combination with bevacizumab as the first‐line treatment in NSCLC patients with brain metastasis harboring EGFR‐activating mutations. However, randomized studies with more patients are required to confirm this finding.In conclusion, osimertinib in combination with bevacizumab is effective and safe in patients with NSCLC and brain metastasis harboring Keke Nie and Youxin Ji designed the study. Ling Zhang and Yunhong You collected the clinical data. Xueli Liu and Fengjun Liu performed the statistical study and preparation of figures. Keke Nie and Youxin Ji drafted this manuscript. All authors read and approved the final manuscript.The authors declare no conflicts of interest.This study was approved by the Ethics Committee of the Affiliated Qingdao Central Hospital of Qingdao University and was performed in compliance with the provisions of the Good Clinical Practice guidelines, the Declaration of Helsinki, and local laws.


To evaluate the effect of parenteral amino acid application in hospitalized hypoalbuminemic dogs.p ≤ 0.05.Medical records of client-owned hypoalbuminemic dogs  were analyzed. Dogs receiving amino acids for only 1–2 days, receiving transfusions or surgery, or <6 months of age were excluded. Dogs were grouped as those receiving intravenous amino acids  over 3 days and longer, and those without additional amino acid treatment . Duration of hospitalization, albumin, and total protein concentrations were compared between groups by Mann–Whitney U test. Course of albumin and total protein concentration was evaluated by Friedman test and Dunn’s multiple comparison test. Significance was set to p < 0.001). Initial albumin concentration was lower in group AA compared to CON (p < 0.001). This difference was no longer present on day 2 (p = 0.134).Dogs in group AA received 10% amino acid solution intravenously over median 4 days (3–11 days). No significant differences regarding survival and adverse effects were observed between groups. Dogs of group AA had significantly longer duration of hospitalization  compared to group CON dogs (median 6 days, 3–24 days; Intravenous application of 10% amino acid solution in hypoalbuminemic dogs can improve albumin concentration after 2 days, but does not influence outcome. It is distributed in the plasma (40%) and the interstitium 60%) . Albumin0% . AlbuHypoalbuminemia is a common disorder in critically ill patients . A serumHypoalbuminemia contributes to life-threatening complications, such as systemic organ failure, pulmonary edema, reduced wound healing, and hypercoagulability .As hypoalbuminemia is often a result of an underlying disease, the treatment should primarily address the underlying process. Measures to increase albumin concentration as well as replacement of albumin function have to be initiated in patients with severe hypoalbuminemia . NaturalIn mechanically ventilated humans, an adequate protein intake adapted on calculations using repeated indirect calorimetry measurement, was found to reduce mortality compared to patietns with daily equal protein intake . The AmeAim of this study was to evaluate the effect of parenteral amino acid application in hospitalized dogs suffering from various diseases causing hypoalbuminemia. This study specifically aimed to evaluate the effect of amino acid application on survival, duration of hospitalization as well as on serum albumin and total protein concentration and time to albumin concentration increase. Correlation of the amino acid dose with increase of serum albumin concentration and adverse effects of amino acid application were investigated in comparison to dogs without amino acid supplementation.2.2.1.Medical records of dogs presented to a university clinic from 2013 to 2019 were reviewed for albumin concentrations ≤25 g/L. Dogs were excluded if they were younger than 6 months, hospitalized for less than 3 days. if albumin was only measured once during hospitalization, if they received plasma, whole blood or human albumin transfusion or if they underwent a surgical procedure (other than placement of feeding tubes) during the examination period. Dogs receiving amino acid infusion for only 1–2 days were also excluded .2.2.® 10%, B. Braun, Melsungen, Germany; group AA) for ≥3 days, and 78 dogs were treated without intravenous amino acids (group CON). Signalment, diagnosis, initial and repeated albumin values, total protein values, systemic inflammatory response syndrome (SIRS) parameters, amount of amino acid application, enteral nutrition by feeding tubes, duration of hospitalization, and survival to discharge were recorded in all dogs. SIRS was diagnosed when ≥2 of the 5 SIRS criteria  were fulfilled as described elsewere . Data were investigated for normality with the D’Agostino & Pearson normality test. Data were presented as median and range. Duration of hospitalization, albumin and total protein concentrations were compared between groups by Mann–Whithney U test. Course of albumin and total protein concentration during hospitalization was evaluated by Friedman test and Dunn’s multiple comparison test. A value of 3.3.1.p = 0.256). The median weight was significantly lower in group AA  compared to the group CON .Median age of dogs in group AA  was not different from those of group CON . Median weight was 14.9 kg (1.0–56.7 kg). Most frequently represented breeds were mixed breed (53), Labrador Retriever (12), Pug (7), Chihuahua (6), Yorkshire Terrier (6), Golden Retriever (5), Bernese Mountain dog (4), Bichon Frisèe (4), German Sheepdog (4), Jack Russel Terrier (4), Dachshund (3), Maltese (3), Cavalier King Spaniel (3), the 44 remaining dogs belonged to various other breeds with less than 3 individuals per breed.p = 0.013; Causes of hypoalbuminemia in the dogs included intestinal loss of albumin (80), renal loss (34), loss through increased vascular permeability (15), reduced synthesis (9), reduced intake (4), and other causes (16) and were different between the study groups (p = 0.831).Dogs in group AA received a median dose of 9 g/100 kcal resting energy requirement (RER)/day  with a total amount of median 31 g/100 kcal RER  of a 10% amino acid solution intravenously over a period of 4 days (3–11 days). Tube feeding was performed in group AA (14/80 dogs) as well as in group CON . A difference in the median positive SIRS criteria at day 0 was neither observed between group AA  and CON , nor between surviving dogs of group AA  and group CON , or between non-surviving dogs of group AA , and group CON .Of the 158 dogs, discharge was documented in 118 dogs. Forty dogs (33.9%) died or were euthanized during hospitalization. No difference in survival rate was found between group AA , and group CON . In surviving dogs, time of hospitalization was also longer in group AA  compared to group CON . However, hospitalization time was not different between non-surviving dogs in group AA  compared to non-surviving dogs in group CON .Median duration of hospitalization in surviving and non-surviving dogs was longer in group AA  compared to group CON . This difference stayed significant at all days with the exceptional day 2. On day 2, serum albumin concentration was not different between both groups . On post hoc Dunn’s multiple comparison test, no daily differences could be identified. In dogs of group CON, serum albumin concentrations were significantly higher on day 5 compared to day 0, to day 1 and to day 2 (p < 0.001).Serum albumin concentrations in group AA were not different on each day of hospitalization compared to day 0 (p < 0.001). This difference was also found on days 1 and 4. On day 2, 3, and 5, the serum total protein concentration was not different between AA and CON (p = 0.002). In group CON serum total protein concentrations were different between the days of hospitalization (p = 0.031), but no difference between specific days could be found during post hoc analysis . Serum albumin increase was higher in CON at day 5 compared to AA (p = 0.042). No difference in change of albumin concentration compared to day 0 was observed between groups at all other study points  compared to group CON, where albumin decreased . On day 2, significantly more dogs in group AA had an albumin increase of p = 0.041).Albumin increased in 54 dogs in group AA and 52 dogs in group CON during hospitalization. On day 2 and day 5, significantly more dogs in group AA had an albumin increase of p = 0.029; p = 0.923).Average daily amino acid dose of 6–9 g/100 kcal RER, but not <6 g/100 kcal RER, and not >9 g/100 kcal RER lead to an increase in albumin difference between day 0 and day 2  solutions without lipids with an osmolality of 920 mosmol/L had a 44% probability to develop phlebitis compared to humans receiving isotonic fluids (26%). The addition of a 10% lipid solution to the TPN decreased osmolarity (712 mosmol/L) and reduced the risk of phlebitis to 22% . Human c4.1.The retrospective nature of the study and lack of uniform criteria for administration of amino acids are the major limitation of this study. The heterogenous study population and different initial albumin levels as day 0 between groups caused a bias in the comparability of the groups. Also, daily albumin concentrations were not available in all dogs. Due to the retrospective nature of the study, calculation of a severity score, such as the APPLE score to prove illness severity of the study groups was not possible.5.This retrospective study showed mild evidence of improvement of albumin concentration in dogs with albumin ≤25 g/L treated with a 10% amino acid infusion compared to dogs not treated with amino acids. Intravenous amino acid application at doses of 6–9 g/100 kcal resting energy requirement/day over 3 days or longer might improve albumin synthesis. Further prospective studies with a standardized study and infusion protocol as well as standardized assessmet of the illness severty and cause of hypoalbuminemia are required to assess the effect of the amino acid infusion on albumin and total protein concentration.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.Ethical review and approval was not required for the animal study because data were only retrospectively analyzed.SS, KH, and RD: conceptualization, investigation, and methodology. SS: data collection and writing original draft. SS and RD: formal analysis. KH and RD: supervision and writing review and editing. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


White matter hyperintensities (WMH), a marker of cerebral small vessel disease and predictor of cognitive decline, are observed at higher rates in persons with HIV (PWH). The use of cocaine, a potent central nervous system stimulant, is disproportionately common in PWH and may contribute to WMH.The sample included of 110 PWH on antiretroviral therapy. Fluid‐attenuated inversion recovery (FLAIR) and T1‐weighted anatomical MRI scans were collected, along with neuropsychological testing. FLAIR images were processed using the Lesion Segmentation Toolbox. A hierarchical regression model was run to investigate predictors of WMH burden .+ T‐cell count was 750. Nearly a third (29%) currently used cocaine regularly, with an average of 23.75 (SD = 20.95) days in the past 90. In the hierarchical linear regression model, cocaine use was a significant predictor of WMH burden (β = .28). WMH burden was significantly correlated with poorer cognitive function (r = −0.27). Finally, higher WMH burden was significantly associated with increased serum concentrations of interferon‐γ‐inducible protein 10 (IP‐10) but lower concentrations of myeloperoxidase (MPO); however, these markers did not differ by COC status.The sample was 20% female and 79% African American with a mean age of 45.37. All participants had persistent HIV viral suppression, and the median CD4WMH burden is associated with poorer cognitive performance in PWH. Cocaine use and CVD risk independently contribute to WMH, and addressing these conditions as part of HIV care may mitigate brain injury underlying neurocognitive impairment. Neurocognitive impairment (NCI) remains a prevalent complication of HIV disease, even in patients with sustained HIV viral suppression, with estimated prevalence rates ranging from 15 to 55% across populations.White matter hyperintensities (WMH), which have been observed at higher rates in PWH,+ T‐cell count below 500 cells/μL (a marker of immunosuppression) was predictive of total WMH load,Aging and cardiovascular disease (CVD) risk factors such as hypertension, diabetes, and smoking are the primary predictors of WMH in the general populationCocaine use is disproportionately common in PWH and may contribute to neuronal injury. The most recent surveillance by the CDC of adults with HIV reported past year prevalence rates of 5% for non‐injection cocaine and 3% for crack cocaine.WMH can be measured quantitatively using non‐invasive magnetic resonance imaging (MRI). Specifically, T2‐weighted fluid‐attenuated inversion recovery (FLAIR) imaging is the industry‐standard sequence for delineating WMHs. The total WMH volume is an important determinant of their clinical relevance.The data in this report were collected as part of a larger project on neuroimaging and immunological features of NCI in PWH.3 or HIV infection of ≥5 years; and sustained viral suppression (plasma HIV RNA < 200 copies/mL) for >1 year. Exclusion criteria included perinatal HIV infection; <9 years of education; BMI > 40; English non‐fluency or illiteracy; severe learning disability; severe head trauma with loss of consciousness >30 min and persistent functional decline; serious neurological disorders not caused by HIV, including stroke; acute CNS infection or chronic CNS infection with residual symptoms; severe mental illness or acute psychiatric symptoms; systemic autoimmune disease; inpatient hospitalization within 30 days; use of an immunomodulatory medication, steroid, or antibiotic within 30 days; contraindications to MRI; and other underlying or uncontrolled medical illness with altered cognition.PWH aged 18–59 years were recruited from infectious diseases clinics in the Raleigh/Durham area of North Carolina, USA. Participants met the following criteria: stable ART for >1 year, nadir CD4 cell count of ≤350 cells/mmThe sample was stratified by cocaine history. A positive history (COC+) was defined by lifetime history of regular use and ≥12 days of use in the past 90 days or a positive urine screen, while a negative history (COC−) was defined as no regular use in the past 10 years, no use in the past 90 days, and a negative urine drug screen. Alcohol, cannabis, and nicotine use were allowed, and controlled substances such as benzodiazepines were permitted if prescribed. For other illicit drugs of abuse, participants were excluded for: a positive urine drug screen; >2 days of use within 30 days; >2 years of regular use within the past 20 years; and any regular use in the past 5 years.The Addiction Severity Index‐Lite (ASI‐L), a semi‐structured interview, was administered to characterize lifetime substance use and associated problems.+ T‐cell counts, current antiretroviral regimen, and most recent plasma HIV viral load. The following clinical diagnoses were coded as present or absent: diabetes, hypertension, hyperlipidemia, and any prior CVD such as coronary artery disease and cardiomyopathy. Body mass index (BMI) was calculated from weight and height measured at last clinical visit. Participants reported frequency of smoking in the past 3 months. As in prior studies,Participants provided a release for the study team to obtain copies of their healthcare records. This information was used to confirm no exclusionary conditions, and to abstract relevant clinical data. HIV disease characteristics included date of diagnosis, nadir and current CD4Attention: Paced Auditory Serial Addition Task‐50—total number correctInformation Processing: Trail Making Test Part A—number of seconds to completionLearning : Hopkins Verbal Learning Test—Revised (HVLT‐R)—total number of words recalled on trials 1–3Memory : HVLT‐R—total number of words recalled on trial 4Executive function: Stroop Color and Word Test interference score—difference between actual and predicted score on the Color‐Word trialVerbal fluency: FAS letter fluency—total number of words generated; and category fluency—total number of animals generatedMotor skills: Grooved Pegboard Test dominant and non‐dominant hand—number of seconds to completionNeuropsychological testing was administered by Bachelor's‐level staff member under the supervision of a licensed clinical psychologist. The 60‐min battery of tests spanned seven domains of cognitive function:T‐scores . Domain T‐scores were computed by averaging the T‐scores for the tests within each domain, and a global T‐score was computed by averaging the 7 domain T‐scores.Using published normative data,3, repetition time [TR] = 8.16 ms, echo time [TE] = 3.18 msec, field of view [FOV] = 256 mm2, 12° flip angle, 168 interleaved slices). Fast‐spin echo T2‐weighted FLAIR images were acquired in oblique axial orientation of the full brain with the following parameters: voxel size = 0.4 mm2 × 5 mm, TR = 10000 msec, TE = 128 msec, TI = 2337 msec, FOV = 220 mm2, 111° flip angle, 27 slices. Total acquisition time for FLAIR was 3:30 min.All MRI scans were performed at Duke University Hospital using a 3.0T GE Discovery MR750 scanner with an 8‐channel head coil. High‐resolution T1‐weighted images were recorded using a spoiled echo sequence (voxel size = 1 mmhttp://www.fil.ion.ucl.ac.uk/spm/software/spm12). This information was combined with the coregistered FLAIR image to provide a lesion belief map for each class of tissue using the automated lesion growth algorithm (LGA) of LST. These maps were thresholded with a lesion probability threshold (κ) to generate an initial binary lesion map that was subsequently grown along voxels that appear hyperintense on the FLAIR image. To define the optimal threshold, T1 and FLAIR images from 11 randomly selected participants were manually segmented by a neuroradiologist, blinded to COC status, to establish ground truth lesion masks. A range of lesion volumes  was included in the training set. These same cases were then segmented at κ = 0.10, κ = 0.20, and κ = 0.30 using the LGA algorithm. Comparing the dice similarity coefficients (DSC) between the manually segmented images and the LGA‐generated lesion belief maps, it was determined that a κ = 0.10 was the optimal threshold . This was confirmed via visual inspection of the segmentation results. The DSC values are in line with other studies with overall low lesion loads, as errors in the segmentation have a greater impact when the lesion load is low.Cortical reconstruction and volumetric segmentation of the T1‐weighted images was done with the FreeSurfer image analysis suite version 6.0 to obtain estimated total intracranial volume (eTIV).Non‐fasting blood samples were collected by peripheral venipuncture immediately following the MRI or on a subsequent day. Plasma and serum were purified using standard procedures, and then, supernatant was pipetted into aliquots and stored at −80°C until time of assay. A selected set of 10 biomarkers representing various components of the inflammatory cascade were measured using commercially available enzyme‐linked immunosorbent assay (ELISA) kits. All analytes were measured in duplicate following manufacturers' instructions. Serum samples were assayed on the Luminex MagPix instrument using commercially available multiplex kits from EMD Millipore  to measure concentrations of IL‐4, IL‐6, IL‐8, IL‐13, and TNFα (Milliplex® Human High Sensitivity T Cell) and MPO (Milliplex® Human CVD Magnetic Bead Panel 2). Single ELISA assays were used for the following analytes: MCPI/CCL2 , IP‐10/CXCL10 , sCD14 , and sCD163 . Laboratory personnel were blinded as to the demographic and COC status of the participants. Four people were missing blood data, and one person was excluded because values were >3 SD above the mean on >3 biomarkers. The values for the cytokines/chemokines were log10 transformed to improve normality.t‐tests, Mann–Whitney U‐tests, chi‐squared tests, and Fisher's exact tests, as appropriate, to compare COC+ and COC− participants. A hierarchical multiple regression model was conducted to identify the independent association of cocaine use on WMH, after accounting for common risk factors. Variables were entered in a series of blocks in the following order: demographics , clinical factors , and substance use . A partial correlation was used to examine the strength of the relationship between WMH load and global cognitive function (T‐score) controlling for age. Finally, a series of analysis of covariance (ancova) models examined the association of WMH burden to each biomarker. Lesion load was the between‐subjects factor defined as minimal, low, moderate, or high, controlling or age. All analyses were conducted in SPSS 28.0.Descriptive statistics were used to characterize the sample on demographic, HIV disease, CVD risk characteristics, and inflammatory biomarkers. We used two‐tailed independent samples The sample included 32 COC+ and 78 COC− participants Table . They weP < 0.05).Participants in the COC+ group had been using cocaine regularly for an average of 18.28 years (SD = 10.70). The majority (66%) reported smoking as their primary route of administration; all others reported nasal administration. In the 90 days prior to enrollment, they had used cocaine on an average of 23.75 days (SD = 20.95). While the overall prevalence of use was high for current alcohol (75%) and cannabis (48%), COC+ participants were significantly more likely than COC− participants to have used these substances . Figure 2(2) = 13.48, P = 0.004). Compared to the COC− group, COC+ participants were more likely to be categorized as high (9% vs. 0%) and moderate (25% vs. 9%) lesion burden and less likely to have minimal lesion burden (28% vs. 44%). Table F  = 3.44, P < 0.001, R2 = 0.26). Step 1 included demographic factors, and age was associated with higher WMH volume. Step 2 added clinical factors. As expected, CVD risk score was associated with higher WMH volume. None of the HIV clinical variables were predictive. Step 3 added the substance use factors, and cocaine was associated with higher WMH volume. In the final model, age, CVD risk, and cocaine use were independent predictors of WMH burden.The median WMH volume was higher for COC+  than for COC−  , suggesting that greater WMH burden is associated with poorer cognitive function  and COC−  groups. In an exploratory analysis, we examined the correlation with domain T‐scores. WMH volume was negatively associated with executive function , learning , and memory .WMH volume was negatively correlated with global ion Fig. . The strancova models comparing biomarker values across WMH categories. Controlling for age, concentrations of IP‐10 and MPO were significantly different across the four categorizations. Specifically, participants who were categorized as having moderate or high lesion burden had the highest concentrations of IP‐10 and the lowest concentration of MPO. However, the group differences did not pass family wise error correction for multiple comparisons. Since IP‐10 and MPO did not differ by COC status, we did not pursue mediation analysis.After accounting for age, the COC group had comparable levels for all of the inflammatory markers except IL‐13 (Table This study investigated the contribution of ongoing cocaine use to WMH in persons with treated HIV disease who had sustained viral suppression. Our principal finding is that cocaine use in PWH was associated with greater WM lesion burden, even after accounting for age and CVD risk factors. Moreover, higher WM lesion burden was correlated with poorer cognitive function, underscoring the role of vascular processes in the development of NCI in PWH independent of COC status. While chronic HIV disease may contribute to the development of cerebral small vessel disease, it appears that age, CVD risk, and cocaine are robust predictors of WMH with high relevance for PWH. Finally, circulating neurotoxic chemokines linked to endothelial functioning were associated with WM lesion burden, suggesting a potential role of inflammatory processes in the pathogenesis of cerebrovascular insults in the context of chronic HIV infection.This study was specifically designed to test the effects of cocaine on health outcomes in PWH, comparing individuals with ongoing cocaine use to those with no history of substantive cocaine use. Our results support the independent role of ongoing cocaine use on burden of WMH. A recent study did not find an association between cocaine use and WMH in PWH, but groups were defined by a past history of cocaine use disorder rather than current use.Consistent with the extensive literature on cerebral small vessel disease,Despite prior reports that HIV disease confers an elevated risk of WMH,At the cellular level, cerebral small vessel disease is believed to result from endothelial dysfunction and subsequent exposure of the arterial vessel wall, making it susceptible to thrombosis.Strengths of this study include our use of comprehensive assessments to characterize the sample and rigorous eligibility criteria related to both substance use and HIV disease to minimize confounds and maximize power. Our study also has some limitations. First, we did not include an HIV‐negative comparison group because the goal of the larger project was to identify neural correlates of NCI in PWH. Second, the sample size of 110 is relatively modest, but the study was designed specifically to isolate the effects of cocaine on HIV‐related outcomes. With clearly delineated groups, our power to detect differences attributed to cocaine use was strengthened. Third, we used a research‐domain automated pipeline for lesion segmentation, as manual delineation of WM lesions is labor intensive with considerable inter‐ and intra‐rater variability.Our principal finding is that PWH who use cocaine have higher WM lesion burden, even after accounting for CVD risk factors, that contribute to cognitive impairment. While the relative burden of WMH remained modest for most participants, these lesions represent only the end of a continuous spectrum of WM injury and neurodegeneration.Conception and design of the study: CSM, RPB, and SLT. Acquisition, analysis, or interpretation of data: CSM, RPB, and CDL. Statistical analysis: CSM, RPB, and SLT. Drafting of the manuscript or figures: CSM, RPB, and SLT. Critical revision of the manuscript: CSM, RPB, SLT, CDL, KA, and MJG. Obtaining of funding: CSM and SLT.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.


Imatinib is the standard first‐line treatment for advanced gastrointestinal stromal tumors (GISTs); however, most patients eventually develop imatinib resistance, leading to considerable clinical challenges. Few direct comparisons have been made between different post‐first‐line therapies on clinical efficacy in advanced GIST following imatinib failure.Databases including PubMed, Embase, Scopus, Google Scholars, and Cochrane Library from inception to February 2023 were retrieved for randomized controlled trials evaluating the clinical efficacy of different post‐first‐line agents for advanced GIST following imatinib failure. Network and conventional meta‐analysis were carried out using Stata/MP 16.0.Ripretinib showed significant improvement in progression‐free survival (PFS) rates from the 2nd to the 12th month compared to placebo, while there was virtually no evidence that the rest active agents had a significant benefit at the 12th month. Masitinib, ripretinib, sunitinib, regorafenib, and pimitespib exhibited significantly longer median PFS than placebo, and pairwise comparisons indicated there were no significant differences among masitinib, ripretinib, and sunitinib. These post‐first‐line agents decreased the risk of disease progression or death by 65%  compared to placebo. Ripretinib and sunitinib came into effect earlier and exhibited more consistent overall survival (OS) rate improvements than masitinib and pimitespib, while pairwise comparisons revealed no significant differences in these four active agents concerning the improvement in OS rate. These post‐first‐line agents decreased the risk of death by 39%  over placebo for advanced GIST following imatinib failure.The active agents in our analysis as post‐first‐line therapies are able to provide superior clinical efficacy, with improved PFS rate and OS rate at certain time points, as well as absolute values of PFS and OS for advanced GIST. Ripretinib might be the optimal recommendation as a post‐first‐line treatment for advanced GIST following imatinib failure.  Most patients diagnosed with GIST are in their 60s, and the gender distribution is fairly equal.GIST patients are usually asymptomatic or have nonspecific symptoms, including abdominal distension, abdominal pain, early satiety, bleeding, fatigue, and palpable mass, which may lead to delay in diagnosis and treatment.Imatinib, a multitargeted TKI targeting KIT and PDGFR, has been widely used as the standard first‐line treatment for advanced GIST, improving the median survival from 1‐to 5 years. However, most patients inevitably develop imatinib resistance often after 20–24 months on imatinib, leading to disease progression and eventually death, and since the approval of imatinib in 2002, there have been no improvements in first‐line therapy for GIST; this has resulted in significant challenges in the clinical management of advanced GIST.There is an unmet medical need for better therapeutic alternatives which can improve the current condition of advanced GIST patients, and to provide more options and clinical benefits for such patients following first‐line imatinib failure.Although an increasing number of studies concerning post‐first‐line treatments for advanced GIST have been conducted, few direct comparisons have been made between these active agents in terms of their treatment efficacy, and no definite conclusions have been drawn concerning their effectiveness. Considering this, we conducted this meta‐analysis incorporating both conventional meta‐analysis and network meta‐analysis, with the aim of summarizing and comparing the clinical efficacy of post‐first‐line therapeutic agents in terms of PFS rate and OS rate at certain time points, as well as absolute values of PFS and OS for advanced GIST after first‐line imatinib therapy failure.22.1We initiated our study by searching the PubMed, Embase, Scopus, Google Scholars, and Cochrane Library databases for randomized controlled trials (RCTs) from their inception to February 2023, using the following keywords: “gastrointestinal stromal tumor” or “GIST”; “advanced” or “metastatic” or “unresectable” or “recurrent,” and “imatinib”. Furthermore, we explored the references of relevant reviews and guidelines for additional candidate publications. Two reviewers screened titles and abstracts independently to verify all potentially eligible publications for full‐text assessment. The final studies included were identified on the basis of the inclusion and exclusion criteria described above. This network meta‐analysis followed the guidelines of PRISMA.2.2The eligible studies were verified according to the following criteria by two experienced investigators individually: (1) the study design must be RCT; (2) patients were diagnosed with advanced GIST and previously experienced first‐line imatinib therapy failure (caused by either disease progression or intolerance); (3) patients received an active agent as post‐first‐line treatment in the research group and an active agent as post‐first‐line treatment or placebo in the control group; (4) the clinical efficacy outcomes were PFS rate and OS rate at certain time points, as well as absolute values of PFS and OS, and atleast one desirable outcome must be reported.2.3Articles were excluded if they were reviews, single‐arm studies, case reports, animal studies, commentaries, conference abstracts, retrospective observational studies, or studies published in a language other than English.2.4p < 0.05 indicated the potential of publication bias. We utilized a network funnel plot to evaluate possible small sample effects in a network meta‐analysis. For each RCT, two investigators independently extracted the data and evaluated the risk of bias according to the Cochrane Collaboration's risk of bias tool.Information extracted included first author, year of publication, study design, comparative treatment regimens, sample size, age, and sex. In addition, HRs and 95% CIs for PFS and OS were extracted. We also extracted the PFS rate and OS rate at certain time points from the PFS and OS curves using GetData software. Publication bias was assessed by using Egger's test, and 2.5p < 0.05 was considered statistically significant.For dichotomous variables, we expressed effects as odds ratios (ORs), while for continuous variables, effects were expressed as weighted mean differences (WMDs) with their 95% CIs. Risks were presented as HRs with 95% CIs. Continuous variables, when presented as the median with the interquartile range, were converted to the mean.33.1The flow diagram presents the details of the literature screening and selection process for the network meta‐analysis Figure . Our sea3.2PFS rates at certain time points have not been extensively explored by previous studies. The present study included a comparison of PFS rate results at certain time points separately; the results are shown in Figure In the meta‐analysis, significant benefit of active agents compared with placebo was confirmed in 2 month , 4 month , 5 month , 6 month , 7 month , 8 month , and 9 month . However, there was no significant improvement in 10 month , 11 month , and 12 month PFS rate . The most effective treatment in terms of PFS rate was found to be ripretinib. In comparison with the placebo, this treatment showed a significant improvement in PFS rate from the 2nd to the 12th month. Moreover, there was virtually no evidence that the rest active agents had significant effect at the 12th month compared with the placebo  and sunitinib  show 2 month OS rate improvements; masitinib , ripretinib  and sunitinib  show 4 month OS rate improvements; masitinib , ripretinib  and sunitinib  show 5 month OS rate improvements; masitinib , ripretinib , pimitespib  and sunitinib  show 6 month OS rate improvements; masitinib , pimitespib , ripretinib  and sunitinib  show 7 month OS rate improvements; pimitespib , ripretinib  and sunitinib  show 8 month OS rate improvements; masitinib , ripretinib , sunitinib , and pimitespib  show 9 month OS rate improvements; and masitinib , ripretinib  and sunitinib  show 10 month OS rate improvements Tables .Overall, masitinib, pimitespib, ripretinib, and sunitinib were observed to provide significantly improved OS rates at certain time points compared with the placebo, while the pairwise comparisons revealed there to be no statistically significant differences among these treatment groups in terms of the OS rate improvement Tables –S9.3.5We analyzed a total of seven3.6p > 0.05) using Egger's test, and there was also no significant publication bias in the HRs of PFS and OS . Additionally, the network funnel plot of pairwise comparisons on PFS and OS rate at certain time points are summarized in Figures No publication bias was identified in the PFS rate ; data curation ; formal analysis ; writing – original draft (lead); writing – review and editing . HU ZHANG: Data curation ; writing – original draft (supporting); writing – review and editing . Mingrong Shu: Data curation ; formal analysis ; writing – review and editing . xingyue wang: Conceptualization ; data curation ; supervision (lead); writing – original draft (supporting); writing – review and editing .The authors disclose no conflicts of interest associated with this work.No ethical approval was required for the current meta‐analysis.Supporting information S1. Supplementary materialClick here for additional data file.


Here, we report that panobinostat and marizomib significantly depleted ATP and NAD+ content, increased mitochondrial permeability and reactive oxygen species generation, and promoted apoptosis in pediatric and adult glioma cell lines at initial treatment. However, resistant cells exhibited increased levels of TCA cycle metabolites, which required for oxidative phosphorylation to meet their bioenergetic needs. Therefore, we targeted glycolysis and the electron transport chain (ETC) with small molecule inhibitors, which displayed substantial efficacy, suggesting that resistant cell survival is dependent on glycolytic and ETC complexes. To verify these observations in vivo, lonidamine, an inhibitor of glycolysis and mitochondrial function, was chosen. We produced two diffuse intrinsic pontine glioma (DIPG) models, and lonidamine treatment significantly increased median survival in both models, with particularly dramatic effects in panobinostat‐ and marizomib‐resistant cells. These data provide new insights into mechanisms of treatment resistance in gliomas.In previous studies, we demonstrated that panobinostat, a histone deacetylase inhibitor, and bortezomib, a proteasomal inhibitor, displayed synergistic therapeutic activity against pediatric and adult high‐grade gliomas. Despite the remarkable initial response to this combination, resistance emerged. Here, in this study, we aimed to investigate the molecular mechanisms underlying the anticancer effects of panobinostat and marizomib, a brain‐penetrant proteasomal inhibitor, and the potential for exploitable vulnerabilities associated with acquired resistance. RNA sequencing followed by gene set enrichment analysis (GSEA) was employed to compare the molecular signatures enriched in resistant compared with drug‐naïve cells. The levels of adenosine 5′‐triphosphate (ATP), nicotinamide adenine dinucleotide (NAD) Brain‐penetrant panobinostat, a histone deacetylase inhibitor, and proteasomal inhibitor marizomib displayed synergistic activity against pediatric and adult high‐grade gliomas. Despite the remarkable initial response to this combination, resistance emerged. The resistant cells exhibited increased levels of TCA cycle metabolites, required to meet their bioenergetic needs. Therefore, we targeted the electron transport chain with small‐molecule inhibitors, which displayed substantial efficacy. Total RNA from four biological replicates was isolated using a Qiagen RNeasy Mini kit  according to the manufacturers' instructions and quantified by measuring the optical density . Using Applied Biosystems  reverse transcription assay kit, 1 μg of total RNA was converted to first‐strand cDNA in a volume of 20 μL containing RNA template, 100 meviously , 47. Thebowtie (v2.3.5.1) algorithm employed in CLC Genomics Workbench 2020 ((Ensemble version 54), Qiagen, CLC Workbench 2020) [2(Transcript per million (TPM) + 1) values < 2 were considered to not be expressed at a sufficient level to permit reliable quantification and were removed. Sample counts were then normalized to counts per million (CPM) and transformed (Log2CPM + 2) using edge r package in the r computing environment. iDEP.94 DESeq2 statistical packages in r were used to identify significant changes in gene expression between experimental conditions [enrichr Package in python [After RNA extractions (as described above), strand‐specific RNA‐Seq library preparation and subsequent sequencing were performed at Novogene Biotech  using standard Illumina protocols carried out on the Illumina NovaSeq 6000 platform as per the manufacturer's instructions ) as described before . Short rch 2020) . Genes wnditions , 51. Sign python , 53, 54.n python , 56.2.85 cells per well and allowed to attach overnight. After treatment with an inhibitor or vehicle for the desired duration, cells were collected, and treated with lysis buffer from the ATP detection kit. The lysate was centrifuged at 12 000 × g for 10 min at 4 °C, and then, the supernatant was moved to a fresh tube. The protein concentration was determined using a Protein Assay Reagent  using BSA as standard. An equal amount of protein was taken; subsequently, the lysate was incubated with enzyme mix and the developer solution (provided in the kit) in 96‐well plates in triplicate. The plate was incubated at room temperature for 30 min in the dark and absorbance was measured at 450 nm by BioTek, Synergy HTX multimode plate reader .The amount of ATP was measured by using an ATP Colorimetric/Fluorometric Assay Kit, according to the manufacturer's protocol . The cells were seeded onto 6‐well plates at 2.5 × 102.9+ content was measured as described previously using the colorimetric assay kit  [Total intracellular NADCA, USA) .2.106 cells per replicate, completed in triplicate. Pellets were flash frozen, and metabolite extraction was completed using ice‐cold 80 : 20 methanol:water with 0.1% formic acid that included internal standards, creatinine‐d3, alanine‐d3, taurine‐d4 and lactate‐d3 at a final concentration of 10 μm. After 3 min of vortexing, the supernatant was cleared of protein by centrifugation at 16 000 × g. Samples of cleared supernatant (2 μL) were injected via a Thermo Vanquish UHPLC and separated over a reversed‐phase Thermo HyperCarb porous graphite column  maintained at 55 °C. For the 20‐min LC gradient, the mobile phase consisted of the following: solvent A (water/0.1% FA) and solvent B (ACN/0.1% FA). The gradient was the following: 0–1 min 1%B, with an increase to 15%B over 5 min, increasing to 98%B over 5 min, holding at 98%B for 5 min, and then equilibration at 1%B for 5 min. The Thermo ID‐X tribrid mass spectrometer was operated in both positive and negative ion mode, scanning in ddMS2 mode (2 μscans) from 70 to 800 m/z at 120 000 resolution with an AGC target of 2e5 for full scan and 2e4 for MS2 scans using HCD fragmentation at stepped 15, 35, 50 collision energies. Source ionization settings were 3.0 and 2.4 kV spray voltage, respectively, for positive and negative modes. Source gas parameters were 35 sheath gas, 12 auxiliary gas at 320 °C, and 8 sweep gas. Calibration was performed prior to analysis using the PierceTM FlexMix Ion Calibration Solutions (Thermo Fisher Scientific). Integrated peak areas were then extracted manually using quan Browser .Drug‐naïve or PM‐resistant cells were collected for steady‐state metabolomics analysis as described before . In brie2.11Pyruvate dehydrogenase (PDH), a mitochondrial multienzyme, that catalyzes the conversion of pyruvate to acetyl‐coenzyme A was assessed using the Pyruvate Dehydrogenase Activity Colorimetric Assay Kit from BioVision  as described elsewhere . Triplic2.12+ metabolome, as defined here, includes dinucleotides, nucleotides, nucleosides, and nucleobases that were measured as described previously [m HEPES, pH 7.1 v/v (buffered ethanol) solution, preheated to 80 °C. Samples were shaken at 100 × g in an 80 °C block for 3 min. Soluble metabolites were separated from particulate by refrigerated microcentrifugation . Though the ethanol‐soluble extract contains all the metabolites of interest, the weight of the particulate can be used to determine the optimized resuspension volume for dried metabolites. Then, both the particulate and soluble metabolites were dried by speed vacuum at 40 °C. For LC–MS analysis, the pellet was resuspended and diluted two‐fold into two different metabolite standards, and 2.5 μL of the resulting material was injected and analyzed. Using internal standards, intracellular metabolites per million cells were reported on a pmol scale.Quantitative NADeviously . In brie2.13Hexokinase (HK) enzymatic activities were assessed by the colorimetric method described by the manufacturer . After adding the reaction mix to the samples, this assay was performed in a 96‐well plate at room temperature at 450 nm.2.14in vivo model was generated using 4‐week‐old NOD‐SCID female mice  injected with DIPG‐013 drug‐naïve or DIPG‐013 PM‐resistant cells. Cells (2.5 × 104) were resuspended in 2 μL of PBS and injected into the pons/midbrain via stereotactic frame  and automated cell injector (Stoelting) over a 4‐min period. Coordinates were as follows from the lambda suture . Mice were injected and separated into two groups: control and experimental. Experimental mice were treated with two rounds of five doses of lonidamine (50 mg·kg−1), for a total of 10 doses. Each round consisted of 1 dose per day, with 2 days of no drug in between rounds. Control mice were dosed with vehicle (5% DMSO in Tris/glycine buffer). Mice were kept in a room at an ambient temperature (73–74°F), with 30% humidity, a dark–light cycle of 14–10 h, in a mouse satellite box with no restrictions on food or water. Mice were monitored for any signs of decline in health daily. Animals losing ≥ 10% body weight or having other symptoms such as hydrocephalus, or neurological duress were sacrificed humanely as defined by IACUC. Survival distribution was determined using Kaplan–Meier analysis [All animal procedures were carried out ethically according to protocols approved by the institutional animal care and use committee (IACUC) at the University of Pittsburgh . The analysis .2.15prism 6.0 graphpad software . To identify differences among subgroups, ANOVA analysis was conducted for multigroup comparisons followed by the post‐hoc Tukey's test. Direct comparisons were conducted using an unpaired two‐tailed Student t‐test.All experiments were performed in biological triplicates unless otherwise stated. The data were expressed as mean ± SD. Statistical analyses were conducted using 33.1in vitro, and dramatic potentiation of cell killing that far exceeded the effects of either agent alone, although these studies incorporated non‐brain‐penetrant agents [m) and marizomib  and their combination on cell proliferation in six glioma cell lines . We used synergyfinder 2.0 [m), marizomib (25 nm) as a single agent or the combination for the indicated duration, and cell death was assessed by Annexin V and propidium iodide (PI) staining by flow cytometry. Although there was a minimal effect of panobinostat and marizomib after 1 day of treatment , the combination of the two agents produced ~ 75%, 90%, and 50% of cell death in KNS42, SJG2, and SF8628, respectively  or the combination for 24 h. After 1 day, the inhibitor was removed and then cells were cultured in an inhibitor‐free medium for 14 additional days. As shown in Fig. To assess whether every cell in the population could re‐enter the cell cycle and grow into a colony (to determine the long‐term effectiveness of the inhibitors), we performed clonogenic growth assays, an 3.22O2 , a critical bioenergetic parameter, is essential for oxidative phosphorylation, ATP synthesis, and cell survival, we used the cationic dye, DiOC6  to measu2O2 Fig.  and supe2O2 Fig. . Pretrea2O2 Fig. , suggest2O2 Fig. . Togethe3.3Although the combination of panobinostat and marizomib holds therapeutic promise, intrinsic or acquired resistance continues to be a major limiting factor for treatment response . To exam2(Fold Change) > 1.5  regulates the oxidation of pyruvate to acetyl‐coenzyme A , a metachromatic dye that binds to the mitochondrial‐specific phospholipid, cardiolipin, where increased fluorescence intensity signal correlates well with increased mitochondrial content [Based on the observation that increased glycolytic and TCA cycle intermediates contributed to the observed resistance in glioma, we hypothesized that increased mitochondrial mass would directly correlate with drug resistance . To addr content  regardle content . As show content . As show+ mediates several metabolic processes that are implicated in drug and radiation resistance in glioma [+ biosynthesis in panobinostat and bortezomib‐resistant glioma cells [+ promotes the proliferation and survival of cancer cells [+ content in drug‐naïve and resistant glioma cell lines [+ level was barely detectable in the short‐term treated cells. By contrast, significant upregulation of NAD+ content was observed in the resistant counterparts. Because NAD+ coenzymes are critical for biosynthetic, bioenergetic, and ROS‐detoxifying processes on which normal and tumor cells depend [+ (QPRT converts quinolinic acid to NaMN), nicotinic acid adenine dinucleotide (NaAD) and second messengers cADPR and ADPR. NAD+ is the substrate for the synthesis of these second messengers that are involved in the regulation of cellular Ca2+ homoeostasis [+ pathways may be necessary to meet the energy demand for cell survival and growth.NADn glioma , 81, 82.ma cells . Becauseer cells , we nextll lines . As showll lines , endogens depend , 85, we eostasis . Indeed,3.5in vitro and in vivo, we examined the impact on the resistance of several inhibitors of oxidative phosphorylation (OXPHOS) and the electron transport chain (ETC). Cells were treated with 2‐DG, an antidiabetic agent that inhibits glycolysis, and with mitochondrial respiratory chain inhibitors IACS‐010759 (inhibitor of complex I), lonidamine (inhibitor of complex II and hexokinase II), antimycin A (inhibitor of complex III), FCCP , and Gboxin  animal model. We selected lonidamine since it has previously been safely applied in vivo and has been shown to interfere with hexokinase activity and mitochondrial energy metabolism by inhibiting glycolysis, triggering dissipation of the mitochondrial transmembrane potential, and increasing ROS generation [in vitro  via intraperitoneal injection with 2 cycles of 5 doses of lonidamine consisting of 1 dose per day, with 2 days intervening between cycle. Kaplan–Meier analysis indicated that lonidamine significantly increased the survival distribution of DIPG‐013‐naïve mice, with a median survival increase from 18 to 25 days (P = 0.0003). Additionally, survival significantly increased in PM‐resistant models with lonidamine treatment, where the median lifespan increased from 19 to 62 days , a glucose analog that inhibits glycolysis, induced a significant reduction in cell viability of PM‐resistant cells compared with drug‐naïve cells. Similarly, treating resistant cells with lonidamine, an inhibitor of hexokinase and complex II (of electron transport chain) , 32, resin vivo context, we chose lonidamine because of its impressive therapeutic activity in our in vitro studies as an inhibitor of both glycolysis and the electron transport chain, and its track record of tolerability in clinical therapeutics [in vitro observations of loss of mitochondrial membrane potential, increased generation of ROS, loss of mitochondrial mass, and cell viability that were particularly dramatic in resistant cells.Then, to test this hypothesis in the apeutics , 109. Ou5The results suggest that physiologically relevant concentrations of panobinostat and marizomib have significant pro‐apoptotic activity in pHGG and aHGG cell lines. However, the challenge presented is neoplastic cells acquire resistance to this combination therapy at clinically achievable doses, limiting the effectiveness of these therapeutic agents. We provide evidence that the emergence of resistance to panobinostat and marizomib is generated due to the highly integrated TCA cycle metabolic network, driving the resistant cells to efficiently replenish the supply of ATP content for survival and growth. In summary, these observations support a critical role for the upregulation of glycolysis and energy metabolism as a mechanism for glioma treatment resistance, which provides a targetable metabolic vulnerability that can be exploited by agents such as lonidamine to enhance glioma cell killing and prolong survival and represents a novel therapeutic strategy that warrants further exploration.The authors declare no conflict of interest.DRP, SA, and IFP conceptualized and designed the study. EPJ, MCR, TAG, MEH, TAM, SJM, UO, DM, and DRP performed the experiments. DRP, EPJ, MCR, TAG, MEH, BJG, SMC, SGW, UO, DM, RD, AM, SA, CB, and IFP analyzed the data. SA, DRP, and IFP supervised the study. All authors participated in data interpretation and manuscript writing, review, and editing. All authors read and approved the final manuscript.https://www.webofscience.com/api/gateway/wos/peer‐review/10.1002/1878‐0261.13427.The peer review history for this article is available at Fig. S1. Enhanced glycolysis and the mitochondrial TCA cycle metabolites are associated with panobinostat‐ and marizomib‐induced resistance.Click here for additional data file.Fig. S2. Quantitative analysis of nucleosides and NAD+ metabolites.Click here for additional data file.Table S1. Interaction landscape of the combination of panobinostat and marizomib for KNS42, SJG2, SF8628, U87, LNZ308, and T98G cell lines.Click here for additional data file.Table S2. Differential gene expression between drug‐naïve vs. PM‐resistant DIPG‐007 and DIPG‐013 cell lines.Click here for additional data file.Data S1. Figure Legends.Click here for additional data file.Data S2. Table Legends.Click here for additional data file.