Post-translational regulation of proto-oncogene ZBTB7A expression by p53 status in cancer cells: HSP90-dependent stabilization vs. p53-KLHL20-ubiquitin proteasomal degradation.

ZBTB7A overexpressed in many human cancers is a major oncogenic driver. ZBTB7A promotes tumorigenesis by regulating transcription of the genes involved in cell survival and proliferation, apoptosis, invasion, and migration/metastasis. One unresolved issue is the mechanism underlying the aberrant overexpression of ZBTB7A in cancer cells. Interestingly, inhibition of HSP90 decreased ZBTB7A expression in a variety of human cancer cells. ZBTB7A interacts with and is stabilized by HSP90. Inhibition of HSP90 by 17-AAG resulted in p53-dependent proteolysis of ZBTB7A via increased p53 expression and upregulation of the CUL3-dependent E3 ubiquitin ligase, KLHL20. Down-regulation of ZBTB7A resulted in the derepression of a major negative regulator of cell cycle progression, p21/CDKN1A. We discovered a new function of p53 regulating ZBTB7A expression through KLHL20-E3 ligase and proteasomal protein degradation system.

One-Dimensional π-d Conjugated Coordination Polymer Intercalated MXene Compound for High-Performance Supercapacitor Electrode.

MXenes possess the characteristics required for high-performance supercapacitors, such as high metallic conductivity and electrochemical activity, but their full potential remains unrealized owing to their tendency to self-restack when fabricated into an electrode. Designing an MXene interlayer with an effective intercalant has, therefore, become an important criterion to alleviate the restacking issue while also synergistically interact with MXene to further improve its electrochemical activity. This study reports the intercalation of 1D π-d conjugated coordination polymer (Ni-BTA) within the Ti3 C2 Tx nanosheet for the fabrication of a highly efficient supercapacitor electrode. Ni-BTA, which consists of a nickel center and 1,2,4,5-benzenetetramine (BTA) organic chain, is uniformly intercalated by direct synthesis on the abundant oxygen terminals on the Ti3 C2 Tx nanosheet surface. The intercalated Ni-BTA acts as an effective charge carrier transportation pathway through its 1D stretched delocalized π-d electrons while participating in pseudocapacitive activity with the Ni centers. As a result, the Ni-BTA/MXene film exhibits excellent rate performance and a gravimetric specific capacitance of 264.4 F g-1 at 5 mV s-1 . This magnitude is retained up to 94.6% after 10 000 cycles. The present study provides insights into the design of MXene interlayers for the fabrication of highly robust and stable supercapacitors.

Risk Factors and Clinical Outcomes of Candidemia Associated With Severe COVID-19.

COVID-19 can cause serious illness requiring multimodal treatment and is associated with secondary infections. Studies have suggested an increased risk of fungal infections, including candidemia following severe COVID-19 though understanding of risk factors and clinical outcomes remains unclear. OBJECTIVES: To describe clinical characteristics, outcomes and risk factors of candidemia among patients hospitalized with severe COVID-19. DESIGN SETTING AND PARTICIPANTS: A multicenter, case-control study of patients with severe COVID-19 was conducted to evaluate risk factors and clinical outcomes in patients who developed candidemia between August 2020 and August 2021. MAIN OUTCOMES AND MEASURES: Chart review evaluating institutional and patient demographics, clinical and mycological characteristics, concomitant interventions (antibiotics, immunosuppressive agents, parenteral nutrition, degree of oxygen support, mechanical ventilation, surgery), treatment regimens, and outcomes (length of stay and discharge disposition). RESULTS: A total of 275 patients were enrolled in the study, including 91 patients with severe COVID-19 and subsequent candidemia and 184 with severe COVID-19 without candidemia. Most patients received antibiotics prior to candidemia episode (93%), while approximately one-quarter of patients received biologic for COVID-19. In-hospital mortality was significantly higher in the cases compared with the controls (68% vs 40%; p < 0.01). Candida albicans was the most common (53%), followed by C. glabrata (19%). Use of central lines, biologic, and paralytics were independent risk factors for candidemia. CONCLUSIONS AND RELEVANCE: Candidemia following COVID-19 infection is a concern that requires clinical consideration and patient monitoring. Risk factors for the development of candidemia in the setting of COVID-19 infection are largely consistent with traditional risk factors for candidemia in hospitalized patients.

Genomic and transcriptomic determinants of response to neoadjuvant therapy in rectal cancer.

The incidence of rectal cancer is increasing in patients younger than 50 years. Locally advanced rectal cancer is still treated with neoadjuvant radiation, chemotherapy and surgery, but recent evidence suggests that patients with a complete response can avoid surgery permanently. To define correlates of response to neoadjuvant therapy, we analyzed genomic and transcriptomic profiles of 738 untreated rectal cancers. APC mutations were less frequent in the lower than in the middle and upper rectum, which could explain the more aggressive behavior of distal tumors. No somatic alterations had significant associations with response to neoadjuvant therapy in a treatment-agnostic manner, but KRAS mutations were associated with faster relapse in patients treated with neoadjuvant chemoradiation followed by consolidative chemotherapy. Overexpression of IGF2 and L1CAM was associated with decreased response to neoadjuvant therapy. RNA-sequencing estimates of immune infiltration identified a subset of microsatellite-stable immune hot tumors with increased response and prolonged disease-free survival.

A Post-Neurosurgical Infection due to KPC-Producing Klebsiella pneumoniae Treated with Meropenem-Vaborbactam: A Case Report.

This case report describes a patient with post-neurosurgical infection and bacteremia caused by Klebsiella pneumoniae carbapenemase-producing Klebsiella pneumoniae. There is limited evidence to guide antibiotic treatment decisions for such infections. The patient was treated with meropenem-vaborbactam (MEV). MEV monotherapy was associated with bacteremia clearance.

Development of Certified Reference Material for Amino Acids in Dried Blood Spots and Accuracy Assessment of Disc Sampling.

To achieve the measurement reliability of amino acids used as diagnostic markers in clinical fields, establishing a reference measurement system is required, in which certified reference materials (CRMs) are an essential step in the hierarchy of measurement traceability. This study describes the development of dried blood spot (DBS) CRMs for amino acid analysis with complete measurement traceability to the International System of Units (SI). Six essential amino acids─proline, valine, isoleucine, leucine, phenylalanine, and tyrosine─were analyzed using isotope-dilution liquid chromatography-mass spectrometry (ID-MS). For minimizing measurement bias and uncertainty overestimation, whole spots with 50 µL of whole blood were adopted in the certification. The between-spot homogeneities by whole spot sampling were lower than 2.1%. The relative expanded uncertainties of the six amino acids in the developed DBS CRMs were lower than 5.7% at 95% confidence. The certified values are traceable to SI through both gravimetric preparation and the primary method in certification, ID-MS. Comparison among DBS testing laboratories revealed discrepancies between the whole spot and disc sampling methods. The actual sampling volume was accurately estimated by weighing, which revealed the possibility of underestimation in routine DBS testing. The candidate CRMs can support the standardization of DBS testing for amino acids through the qualification and validation of many kinds of measurement procedures to compensate the measurement bias caused by matrix-specific sampling error.

KRAS Mutants Upregulate Integrin ß4 to Promote Invasion and Metastasis in Colorectal Cancer.

KRAS mutation in colorectal cancer is associated with aggressive tumor behavior through increased invasiveness and higher rates of lung metastases, but the biological mechanisms behind these features are not fully understood. In this study, we show that KRAS-mutant colorectal cancer upregulates integrin α6ß4 through ERK/MEK signaling. Knocking-out integrin ß4 (ITGB4) specifically depleted the expression of integrin α6ß4 and this resulted in a reduction in the invasion and migration ability of the cancer cells. We also observed a reduction in the number and area of lung metastatic foci in mice that were injected with ITGB4 knockout KRAS-mutant colorectal cancer cells compared with the mice injected with ITGB4 wild-type KRAS-mutant colorectal cancer cells, while no difference was observed in liver metastases. Inhibiting integrin α6ß4 in KRAS-mutant colorectal cancer could be a potential therapeutic target to diminish the KRAS-invasive phenotype and associated pulmonary metastasis rate. IMPLICATIONS: Knocking-out ITGB4, which is overexpressed in KRAS-mutant colorectal cancer and promotes tumor aggressiveness, diminishes local invasiveness and rates of pulmonary metastasis.

An immunocompetent rectal cancer model to study radiation therapy.

We describe a mouse model of rectal cancer (RC) involving rapid tumor organoid engraftment via orthotopic transplantation in an immunocompetent setting. This approach uses simple mechanical disruption to allow engraftment, avoiding the use of dextran sulfate sodium. The resulting RC tumors invaded from the mucosal surface and metastasized to distant organs. Histologically, the tumors closely resemble human RC and mirror remodeling of the tumor microenvironment in response to radiation. This murine RC model thus recapitulates key aspects of human RC pathogenesis and presents an accessible approach for more physiologically accurate, preclinical efficacy studies.

Linking the Child Behavior Checklist to the Strengths and Difficulties Questionnaire.

The Child Behavior Checklist (CBCL) and Strengths and Difficulties Questionnaire (SDQ) both measure emotional and behavioral problems in children and adolescents, and scores on the two instruments are highly correlated. When administrative needs compel practitioners to change the instrument used or data from the two measures are combined to perform pooled analyses, it becomes necessary to compare scores on the two instruments. To enable such comparisons, we score linked three domains (Internalizing, Externalizing, and Total Problems) of the CBCL and SDQ in three age groups spanning 2-17 years. After assessing linking assumptions, we compared item response theory (IRT) and equipercentile linking methods to identify the most statistically justifiable link, ultimately selecting equipercentile linking with loglinear smoothing due to its minimal bias and the ability to link raw SDQ scores with both T-scores and raw scores from the CBCL. We derived crosswalk conversion tables to convert scores on one measure to the metric of the other and discuss the use of these tables in research and practice. (PsycInfo Database Record (c) 2022 APA, all rights reserved).

Significant Domains of Life Satisfaction That Affect Suicidal Behavior in Adolescents.

OBJECTIVES: Adolescent suicide is a serious national issue in Korea. Recently, life satisfaction has been recognized as a major factor related to this issue. The main purpose of this study was to identify the domains of life satisfaction that affect suicidal behavior in adolescence. METHODS: Data were collected from eight middle schools in Incheon, Korea. A total of 1297 students answered questions regarding their demographic characteristics, happiness, self-related life satisfaction domains (appearance, leisure time, physical health, and mental health), depressive symptoms, and suicidal behavior. RESULTS: In the Spearman correlation analysis, female sex, perceived socioeconomic status (SES), happiness, and all four self-related satisfaction scores showed significant correlations with depression and suicidality. Multivariate regression analysis suggested that suicidality was significantly affected by perceived SES, satisfaction with appearance, mental health satisfaction, and depression. Finally, depression was identified as a partial mediator of the association between mental health satisfaction and suicidality, and a complete mediator of the association between female sex and suicidality. CONCLUSION: Perceived SES, satisfaction with appearance, and mental health satisfaction significantly affected students' suicidality, with or without the effect of depression. Health authorities, educators, and family members must be aware of this to identify adolescents at suicide risk earlier.

An antibody against L1 cell adhesion molecule inhibits cardiotoxicity by regulating persistent DNA damage.

Targeting the molecular pathways underlying the cardiotoxicity associated with thoracic irradiation and doxorubicin (Dox) could reduce the morbidity and mortality associated with these anticancer treatments. Here, we find that vascular endothelial cells (ECs) with persistent DNA damage induced by irradiation and Dox treatment exhibit a fibrotic phenotype (endothelial-mesenchymal transition, EndMT) correlating with the colocalization of L1CAM and persistent DNA damage foci. We demonstrate that treatment with the anti-L1CAM antibody Ab417 decreases L1CAM overexpression and nuclear translocation and persistent DNA damage foci. We show that in whole-heart-irradiated mice, EC-specific p53 deletion increases vascular fibrosis and the colocalization of L1CAM and DNA damage foci, while Ab417 attenuates these effects. We also demonstrate that Ab417 prevents cardiac dysfunction-related decrease in fractional shortening and prolongs survival after whole-heart irradiation or Dox treatment. We show that cardiomyopathy patient-derived cardiovascular ECs with persistent DNA damage show upregulated L1CAM and EndMT, indicating clinical applicability of Ab417. We conclude that controlling vascular DNA damage by inhibiting nuclear L1CAM translocation might effectively prevent anticancer therapy-associated cardiotoxicity.

Proto-oncoprotein Zbtb7c and SIRT1 repression: implications in high-fat diet-induced and age-dependent obesity.

Zbtb7c is a proto-oncoprotein that controls the cell cycle and glucose, glutamate, and lipid metabolism. Zbtb7c expression is increased in the liver and white adipose tissues of aging or high-fat diet-fed mice. Knockout or knockdown of Zbtb7c gene expression inhibits the adipocyte differentiation of 3T3-L1 cells and decreases adipose tissue mass in aging mice. We found that Zbtb7c was a potent transcriptional repressor of SIRT1 and that SIRT1 was derepressed in various tissues of Zbtb7c-KO mice. Mechanistically, Zbtb7c interacted with p53 and bound to the proximal promoter p53RE1 and p53RE2 to repress the SIRT1 gene, in which p53RE2 was particularly critical. Zbtb7c induced p53 to interact with the corepressor mSin3A-HADC1 complex at p53RE. By repressing the SIRT1 gene, Zbtb7c increased the acetylation of Pgc-1α and Pparγ, which resulted in repression or activation of Pgc-1α or Pparγ target genes involved in lipid metabolism. Our study provides a molecular target that can overexpress SIRT1 protein in the liver, pancreas, and adipose tissues, which can be beneficial in the treatment of diabetes, obesity, longevity, etc.

Real-World Observational Review of Andexanet Alfa Prescribing and Utilization Outcomes at a Community Teaching Hospital.

OBJECTIVES: Andexanet alfa is the first approved antidote in the management of life-threatening bleeds in patients treated with Xa inhibitors. The ANNEXA-4 study was successful in reducing factor Xa levels during time of administration but lacked correlation to improved patient outcomes. Given its novel mechanism of action, U.S. boxed warning, cost of up to $58,000 per dose, and limited efficacy data compared with standard of care, hospitals are faced with a dilemma with its addition to formulary and process for ensuring optimized use. The objective of this study was to evaluate adherence to institution restriction criteria and the clinical outcomes of treatment for patients for whom andexanet alfa is requested. DESIGN: Retrospective cohort study of andexanet alfa requests within a 12-month time period. SETTING: A 600-bed community teaching hospital. PATIENTS: Patients whom pharmacists received request for dispensing andexanet alfa. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Quality outcomes reviewed compliance to restriction criteria. Clinical outcomes evaluated use of adjunctive blood products, ICU length of stay, hospital length of stay, and hospital mortality. Safety outcomes evaluated incidence of thrombotic events.Andexanet alfa was requested for 16 patients from November 2018 to November 2019. It was administered in nine patients, with compliance to restriction criteria of 66.6%, average ICU length of stay 5.6 days, hospital length of stay 8.6 days, hospital mortality in 44.4%, and thrombotic events in 33.3%. Orders were rejected in seven patients with compliance to restriction criteria of 100%, ICU length of stay 3.2 days, hospital length of stay 5.5 days, hospital mortality in 14%, and thrombotic events in 14%. CONCLUSIONS: A greater rate of adverse effects and mortality was identified with the use of andexanet alfa compared with clinical trials. This is potentially due to its use in a more severely ill patient population and lack of adherence to restriction criteria.

KRAS mutant rectal cancer cells interact with surrounding fibroblasts to deplete the extracellular matrix.

Somatic mutations in the KRAS oncogene are associated with poor outcomes in locally advanced rectal cancer but the underlying biologic mechanisms are not fully understood. We profiled mRNA in 76 locally advanced rectal adenocarcinomas from patients that were enrolled in a prospective clinical trial and investigated differences in gene expression between KRAS mutant (KRAS-mt) and KRAS-wild-type (KRAS-wt) patients. We found that KRAS-mt tumors display lower expression of genes related to the tumor stroma and remodeling of the extracellular matrix. We validated our findings using samples from The Cancer Genome Atlas (TCGA) and also by performing immunohistochemistry (IHC) and immunofluorescence (IF) in orthogonal cohorts. Using in vitro and in vivo models, we show that oncogenic KRAS signaling within the epithelial cancer cells modulates the activity of the surrounding fibroblasts in the tumor microenvironment.

Posterior repair versus no posterior repair for posterior vaginal wall prolapse resolved under simulated apical support at the time of native tissue apical suspension.

INTRODUCTION AND HYPOTHESIS: The aim of this study was to evaluate the impact of an adjuvant posterior repair (PR) on treatment outcomes of native tissue apical suspension. MATERIALS AND METHODS: This retrospective cohort study included 194 women who underwent iliococcygeus or uterosacral ligament suspension with or without PR for Pelvic Organ Prolapse Quantification (POPQ) stage 2-4 posterior vaginal wall prolapse that resolved under simulated preoperative apical support and who completed a 1-year follow-up. The primary outcome was composite surgical failure defined as the presence of vaginal bulge symptoms, descent of the vaginal apex more than one-third of the way into the vaginal canal (apical recurrence), anterior or posterior vaginal wall descent beyond the hymen (anterior or posterior recurrence), or retreatment for prolapse. Secondary outcomes included anatomical outcomes, perioperative outcomes, obstructed defecation, dyspareunia, and adverse events. RESULTS: One hundred thirty women underwent concomitant PR, and 64 did not. Surgical failure rates were significantly higher in the group not receiving PR than in the group receiving PR (29.7% vs. 12.3%, p < 0.01). Anatomically, anterior and apical recurrence was more common in the group not receiving PR (p < 0.05). Concomitant PR was associated with a longer operating time and more blood loss (p < 0.01). However, there were few adverse events related to PR, and the rates of de novo obstructed defecation and dyspareunia were low in both groups, with no significant difference between the groups. CONCLUSION: Concomitant PR at the time of native tissue apical suspension may reduce the recurrence of symptomatic anterior and apical prolapse without significant morbidity.

Variables Affecting Attitudes Toward Suicide Among the Adolescent Population.

Past research has supported the positive association between prosuicide attitudes and suicidal behavior. The aim of the present study was to determine the factor structure of adolescents' attitudes toward suicide and to explore correlates associated with their attitudes. A questionnaire was distributed to 1292 adolescents at eight middle schools to assess their demographic information, clinical variables, and attitudes toward suicide. After factor analysis, we reached a four-factor solution of the attitudes toward suicide. Significantly more females, nonreligious adolescents, those with a lower socioeconomic status, those with higher levels of depressive symptoms, and those with a history of suicidal ideation/plans had more understanding attitudes toward suicide. Depressive adolescents were also more permissive and believed that suicides were unpreventable and that loneliness led to suicide. In conclusion, adolescents' attitudes toward suicide were significantly associated with not only various sociodemographic correlates but also the severity of depressive symptoms and their own experiences of suicidality.

Pharmacologic Inhibition of HIF-1α Attenuates Radiation-Induced Pulmonary Fibrosis in a Preclinical Image Guided Radiation Therapy.

PURPOSE: Radiation-induced pulmonary fibrosis (RIPF) is a long-term side effect of thoracic radiation therapy. Hypoxia-induced vascular endothelial mesenchymal transition (EndMT) can occur during the development of RIPF. Here, we examined the direct contribution of endothelial HIF-1α (EC-HIF1α) on RIPF. METHODS AND MATERIALS: An inducible Cre-lox-mediated endothelial Hif1a deletion mouse line was used to evaluate the potential of HIF-1α inhibition to suppress RIPF. To evaluate the effects of a pharmacologic HIF-1α inhibitor on RIPF after image guided radiation therapy (IGRT) for spontaneous lung adenocarcinoma, we generated conditional tdTomato; K-RasG12D; and p53 flox/flox mice to facilitate tracking of tumor cells expressing tdTomato. RESULTS: We found that vascular endothelial-specific HIF-1α deletion shortly before radiation therapy inhibited the progression of RIPF along with reduced EndMT, whereas prolonged deletion of endothelial HIF-1α before irradiation did not. Moreover, we revealed that postirradiation treatment with the novel HIF-1α inhibitor, 2-methoxyestradiol (2-ME) could efficiently inhibit RIPF and EndMT. In addition, IGRT using primary mouse models of non-small cell lung cancer showed that combined treatment of 2-ME with ablative high-dose radiation therapy efficiently inhibited RIPF and the growth of both multifocal and single tumors, concomitantly reducing radiation-induced EndMT of normal as well as tumor regions. CONCLUSION: These results suggest that a negative regulator of HIF-1α-mediated EndMT, such as 2-ME, may serve as a promising inhibitor of RIPF in radiation therapy.

Association between Perceived Parenting Style and Adolescents' Attitudes toward Suicide.

OBJECTIVES: Adolescent suicide, a major cause of adolescent death, is affected by various factors, including attitudes toward suicide. This study investigated the association between parenting style and adolescents' attitudes toward suicide and the mediating role of attitude toward suicide between parenting style and suicidal ideation. METHODS: We surveyed 1,071 adolescents from eight middle schools in Incheon, Korea. The survey included sociodemographic information, attitudes toward suicide, perception of parenting style, depression severity, and suicidality. RESULTS: Students in the authoritarian parenting group had a more permissive attitude toward suicide compared with the democratic and permissive parenting groups. These students considered that suicide is justified in certain situations and that choosing suicide is an individual's right. They also had a negative attitude toward talking about suicide or intervening in others' suicide. This association remained statistically significant after adjusting for the impact of confounding factors that could affect attitudes toward suicide, except for suicidal processes and preparedness to prevent suicide. In the mediation analysis, we observed that some factors of the attitudes toward suicide mediated between authoritarian parenting attitudes and suicidal ideation, namely, suicide as a right, preventability, suicide as normal/common, preparedness to prevent suicide, and resignation. CONCLUSION: This study revealed the significant impact of parenting style on children's attitudes toward suicide. Educating parents about the appropriate parenting attitudes-sympathetic and rational-can help prevent youth suicide.

Cell fate decisions by c-Myc depend on ZBTB5 and p53.

The oncoprotein, c-Myc, not only promotes cell proliferation, but can also induce or sensitize cells to apoptosis. However, how c-Myc decides cell fate and which c-Myc downstream target genes are involved remain unknown. Previously, we showed that ZBTB5 (zinc finger and BTB domain-containing 5) is a proto-oncogene that stimulates cell proliferation. ZBTB5 represses p21/CDKN1A by competing with p53 and recruiting corepressor histone deacetylase complexes. Herein, we found that c-Myc directly activates the transcription of ZBTB5. In the absence of p53, ZBTB5 is acetylated at K597 by interacting with p300, and activates transcription of NOXA, which induces apoptosis. In contrast, in the presence of p53, ZBTB5 interacts with p53 and acetylation at ZBTB5 K597 is blocked. ZBTB5 without K597 acetylation interacts with mSin3A/HDAC1 to repress p21/CDKN1A transcription and promote cell proliferation. Cell fate decisions by c-Myc depend on ZBTB5, p53 and p300, and acetylation of ZBTB5 K597.

KLHL4, a novel p53 target gene, inhibits cell proliferation by activating p21WAF/CDKN1A.

KLHL4 is a member of the KLHL protein family, many of whom bind the Cul3 E3 ligase, and mediate the ubiquitination of interacting proteins. The KLHL4 gene, localized on the X chromosome, associates with a disorder known as X-linked cleft palate (CPX). However, the biological functions of KLHL4 are largely unknown. In this study, microarray analysis of HEK293A embryonic kidney cells, expressing ectopic p53, showed a 3-fold increase of KLHL4 mRNA. Moreover, both KLHL4 mRNA and protein expression were elevated by p53 or DNA damage, suggesting that KLHL4 might be a p53 target gene. We also found that KLHL4 activates transcription of p21WAF/CDKN1A, a p53 target gene encoding a major negative regulator of the cell-cycle. KLHL4 interacted with p53 to increase its binding to p53 response element of the p21WAF/CDKN1A gene, resulting in transcriptional upregulation. Furthermore, we observed that KLHL4 can interact with the Cul3 ubiquitin ligase, to possibly play a role in ubiquitin-mediated proteasomal degradation, and Klhl4 knocked-out MEF mouse embryonic fibroblasts proliferated faster than WT MEF cells. These results suggest that KLHL4 upregulation by p53 may inhibit cell proliferation, by activating p21WAF/CDKN1A.