Venetoclax resistance leads to broad resistance to standard-of-care anti-MM agents, but not to immunotherapies.

Venetoclax is the first example of personalized medicine for multiple myeloma (MM), with meaningful clinical activity as a monotherapy and in combination in myeloma patients harboring the t(11:14) translocation. However, despite the high response rates and prolonged PFS, a significant proportion of patients eventually relapse. Here, we aimed to study adaptive molecular responses after the acquisition of venetoclax resistance in sensitive t(11:14) MM cell models. We therefore generated single-cell venetoclax-resistant t(11:14) MM cell lines and investigated the mechanisms contributing to resistance as well as the cells' sensitivity to other treatments. Our data suggests that acquired resistance to venetoclax is characterized by reduced mitochondrial priming and changes in BCL-2 family proteins' expression in MM cells, conferring broad resistance to standard-of-care anti-myeloma drugs. However, our results show that the resistant cells are still sensitive to immunotherapeutic treatments, highlighting the need to consider appropriate sequencing of these treatments following venetoclax-based regimens.

Genome wide nucleosome landscape shapes 3D chromatin organization.

The hierarchical chromatin organization begins with formation of nucleosomes, which fold into chromatin domains punctuated by boundaries and ultimately chromosomes. In a hierarchal organization, lower levels shape higher levels. However, the dependence of higher-order 3D chromatin organization on the nucleosome-level organization has not been studied in cells. We investigated the relationship between nucleosome-level organization and higher-order chromatin organization by perturbing nucleosomes across the genome by deleting Imitation SWItch (ISWI) and Chromodomain Helicase DNA-binding (CHD1) chromatin remodeling factors in budding yeast. We find that changes in nucleosome-level properties are accompanied by changes in 3D chromatin organization. Short-range chromatin contacts up to a few kilo-base pairs decrease, chromatin domains weaken, and boundary strength decreases. Boundary strength scales with accessibility and moderately with width of nucleosome-depleted region. Change in nucleosome positioning seems to alter the stiffness of chromatin, which can affect formation of chromatin contacts. Our results suggest a biomechanical "bottom-up" mechanism by which nucleosome distribution across genome shapes 3D chromatin organization.

ABL1 kinase plays an important role in spontaneous and chemotherapy-induced genomic instability in multiple myeloma.

ABSTRACT: Genomic instability contributes to cancer progression and is at least partly due to dysregulated homologous recombination (HR). Here, we show that an elevated level of ABL1 kinase overactivates the HR pathway and causes genomic instability in multiple myeloma (MM) cells. Inhibiting ABL1 with either short hairpin RNA or a pharmacological inhibitor (nilotinib) inhibits HR activity, reduces genomic instability, and slows MM cell growth. Moreover, inhibiting ABL1 reduces the HR activity and genomic instability caused by melphalan, a chemotherapeutic agent used in MM treatment, and increases melphalan's efficacy and cytotoxicity in vivo in a subcutaneous tumor model. In these tumors, nilotinib inhibits endogenous as well as melphalan-induced HR activity. These data demonstrate that inhibiting ABL1 using the clinically approved drug nilotinib reduces MM cell growth, reduces genomic instability in live cell fraction, increases the cytotoxicity of melphalan (and similar chemotherapeutic agents), and can potentially prevent or delay progression in patients with MM.

Assessment of Adverse Childhood Experiences in the South Bronx on the Risk of Developing Chronic Disease as Adults.

Background Adverse childhood experiences (ACEs) have a negative impact on health outcomes. Using a cross-sectional study design, our objective was to identify the prevalence of ACEs among residents of South Bronx and the increased relationship between such childhood stressors and the prevalence of both chronic disease and modifiable high-risk behavior in adulthood. Methods We recruited patients from a hospital-based, adult primary care clinic in the metropolitan area of the South Bronx. A prospectively designed, observational study recruited patients in a consecutive fashion to conduct a cross-sectional survey between September 2017 and January 2018. The demographic representation comprises a low socioeconomic sector of urban New York City, with low education and immigrant population. A modified ACE questionnaire that included nine ACE categories (Physical Abuse, Sexual Abuse, Household Substance Abuse, Separation from Parents, Incarcerated Household Member, Parental Separation/Divorce, and Bullying) in addition to questions on demographics, high-risk behavior, and diagnosis of chronic disease. Our primary objective was to gather the incidence of ACEs organized by domains. Secondary objectives were to demonstrate any expected increase (as odds ratios (ORs)) in chronic disease or maladaptive social habits when compared to patients with no ACEs within the cohort. The OR for the associations was calculated with logistic regression. Individual logistic regression models for each chronic disease, high-risk behavior, and demographics were used to measure the exposure response of the nine ACE categories.  Results A total of 454 patients completed the survey. The average age was 53.1±14.2 years, and females were 49% of the sample. Hispanics were at 61% followed by Blacks at 34%. Participants reported high-risk behavior at 24%, had a high prevalence of chronic illness (82%), and had ACE events at 70%. We found a significant relationship between ACE events and having a chronic disease diagnosis and engagement in high-risk behavior with higher odds of reporting chronic illnesses among participants with exposure to childhood stressors (OR 1.26, 95% confidence interval 1.1-1.5, p=0.002). Of the nine ACE categories, many were independently associated with one or more chronic diseases in adulthood. Conclusion According to our survey data, ACE events in our patient population were more prevalent (30% with four or more exposures), higher than the proposed average of one out of six Americans with four or more exposures nationally according to national statistics. These childhood stressors appeared to have a strong association with the development of high-risk behavior and chronic illnesses.

A Case of Locally Advanced Fibrosarcoma in a Young Male.

Fibrosarcoma is a rare, highly malignant tumor that develops from uncontrolled overgrowth of fibroblastic cells. It may start as a painless lump or swelling under the skin. But as it grows, it can push surrounding structures - organs, muscles, nerves, or blood vessels - and lead to pain and tenderness. The treatment of fibrosarcoma depends on several factors such as size, grade, location of primary tumor, extent of spread, age, and general health condition of the patient. The main treatment is surgical removal of the primary tumor with wide-margin excision and amputation if localized in the limb. Prior to surgical intervention, radiotherapy can be applied to reduce the tumor size or following surgery to lower the risk of recurrence. Chemotherapy is indicated in cases of metastasis. Unfortunately, the prognosis of fibrosarcoma is not favorable. For high-grade fibrosarcoma, the five-year survival rate is around 30% and for low-grade fibrosarcoma, it is 50-80%, with recurrence in the first two to five years post-surgery. We encountered a case of high-grade fibrosarcoma with aggressive growth in a 36-year-old male, requiring above-knee amputation.

Unlocking Endocrown Restoration Expertise Among Dentists: Insights from a Multi-Center Cross-Sectional Study.

BACKGROUND This study, employing an online questionnaire, aimed to assess and contrast the awareness, knowledge, and perceptions of dentists in Saudi Arabia (SA) regarding endocrowns (EC) as post-endodontic restorations with those of dentists educated in various other countries. MATERIAL AND METHODS We conducted a cross-sectional survey among dental interns and practicing dentists in government facilities, private dental centers, and dental colleges in SA, including participants of diverse nationalities. We disseminated validated, closed-ended questionnaires through WhatsApp via Google Forms. The Chi-square test was applied to assess associations between categorical variables, with a P-value of ≤0.05 indicating statistical significance. RESULTS The majority of participants (61.2%) indicated that EC restorations are best suited for molar teeth. Furthermore, 69.6% asserted that the primary objective of employing EC is to accomplish minimally invasive preparations while preserving the existing tooth structure. Among the responses, 68.3% pinpointed debonding of ECs as a significant cause of failure. Notably, substantial differences were observed in responses concerning the knowledge or practice of EC across various factors such as gender, educational attainment, country of graduation, and workplace. CONCLUSIONS The findings reveal a comparatively low adoption of ECs among the participants, irrespective of experience or country of education. This underscores the need for incorporating ECs into dental curricula through theoretical and clinical discussions or considering them as a subject for post-graduate continuing education programs.

Elevated APE1 Dysregulates Homologous Recombination and Cell Cycle Driving Genomic Evolution, Tumorigenesis, and Chemoresistance in Esophageal Adenocarcinoma.

BACKGROUND & AIMS: The purpose of this study was to identify drivers of genomic evolution in esophageal adenocarcinoma (EAC) and other solid tumors. METHODS: An integrated genomics strategy was used to identify deoxyribonucleases correlating with genomic instability (as assessed from total copy number events in each patient) in 6 cancers. Apurinic/apyrimidinic nuclease 1 (APE1), identified as the top gene in functional screens, was either suppressed in cancer cell lines or overexpressed in normal esophageal cells and the impact on genome stability and growth was monitored in vitro and in vivo. The impact on DNA and chromosomal instability was monitored using multiple approaches, including investigation of micronuclei, acquisition of single nucleotide polymorphisms, whole genome sequencing, and/or multicolor fluorescence in situ hybridization. RESULTS: Expression of 4 deoxyribonucleases correlated with genomic instability in 6 human cancers. Functional screens of these genes identified APE1 as the top candidate for further evaluation. APE1 suppression in EAC, breast, lung, and prostate cancer cell lines caused cell cycle arrest; impaired growth and increased cytotoxicity of cisplatin in all cell lines and types and in a mouse model of EAC; and inhibition of homologous recombination and spontaneous and chemotherapy-induced genomic instability. APE1 overexpression in normal cells caused a massive chromosomal instability, leading to their oncogenic transformation. Evaluation of these cells by means of whole genome sequencing demonstrated the acquisition of changes throughout the genome and identified homologous recombination as the top mutational process. CONCLUSIONS: Elevated APE1 dysregulates homologous recombination and cell cycle, contributing to genomic instability, tumorigenesis, and chemoresistance, and its inhibitors have the potential to target these processes in EAC and possibly other cancers.

High-dose melphalan treatment significantly increases mutational burden at relapse in multiple myeloma.

High-dose melphalan (HDM) improves progression-free survival in multiple myeloma (MM), yet melphalan is a DNA-damaging alkylating agent; therefore, we assessed its mutational effect on surviving myeloma cells by analyzing paired MM samples collected at diagnosis and relapse in the IFM 2009 study. We performed deep whole-genome sequencing on samples from 68 patients, 43 of whom were treated with RVD (lenalidomide, bortezomib, and dexamethasone) and 25 with RVD + HDM. Although the number of mutations was similar at diagnosis in both groups (7137 vs 7230; P = .67), the HDM group had significantly more mutations at relapse (9242 vs 13 383, P = .005). No change in the frequency of copy number alterations or structural variants was observed. The newly acquired mutations were typically associated with DNA damage and double-stranded breaks and were predominantly on the transcribed strand. A machine learning model, using this unique pattern, predicted patients who would receive HDM with high sensitivity, specificity, and positive prediction value. Clonal evolution analysis showed that all patients treated with HDM had clonal selection, whereas a static progression was observed with RVD. A significantly higher percentage of mutations were subclonal in the HDM cohort. Intriguingly, patients treated with HDM who achieved complete remission (CR) had significantly more mutations at relapse yet had similar survival rates as those treated with RVD who achieved CR. This similarity could have been due to HDM relapse samples having significantly more neoantigens. Overall, our study identifies increased genomic changes associated with HDM and provides rationale to further understand clonal complexity.

A MIR17HG-derived long noncoding RNA provides an essential chromatin scaffold for protein interaction and myeloma growth.

Long noncoding RNAs (lncRNAs) can drive tumorigenesis and are susceptible to therapeutic intervention. Here, we used a large-scale CRISPR interference viability screen to interrogate cell-growth dependency to lncRNA genes in multiple myeloma (MM) and identified a prominent role for the miR-17-92 cluster host gene (MIR17HG). We show that an MIR17HG-derived lncRNA, named lnc-17-92, is the main mediator of cell-growth dependency acting in a microRNA- and DROSHA-independent manner. Lnc-17-92 provides a chromatin scaffold for the functional interaction between c-MYC and WDR82, thus promoting the expression of ACACA, which encodes the rate-limiting enzyme of de novo lipogenesis acetyl-coA carboxylase 1. Targeting MIR17HG pre-RNA with clinically applicable antisense molecules disrupts the transcriptional and functional activities of lnc-17-92, causing potent antitumor effects both in vitro and in vivo in 3 preclinical animal models, including a clinically relevant patient-derived xenograft NSG mouse model. This study establishes a novel oncogenic function of MIR17HG and provides potent inhibitors for translation to clinical trials.

Machine learning classification reveals robust morphometric biomarker of glial and neuronal arbors.

Neurons and glia are the two main cell classes in the nervous systems of most animals. Although functionally distinct, neurons and glia are both characterized by multiple branching arbors stemming from the cell bodies. Glial processes are generally known to form smaller trees than neuronal dendrites. However, the full extent of morphological differences between neurons and glia in multiple species and brain regions has not yet been characterized, nor is it known whether these cells can be reliably distinguished based on geometric features alone. Here, we show that multiple supervised learning algorithms deployed on a large database of morphological reconstructions can systematically classify neuronal and glial arbors with nearly perfect accuracy and precision. Moreover, we report multiple morphometric properties, both size related and size independent, that differ substantially between these cell types. In particular, we newly identify an individual morphometric measurement, Average Branch Euclidean Length that can robustly separate neurons from glia across multiple animal models, a broad diversity of experimental conditions, and anatomical areas, with the notable exception of the cerebellum. We discuss the practical utility and physiological interpretation of this discovery.

The Observation of Pediatric Skull Fractures Without an Associated Brain Injury in a Non-Trauma Center.

Introduction Young children experiencing head trauma are prone to skull fractures. Pediatric skull fractures are distinct from adults as they have a greater capacity to undergo remodeling. The objective of this study was to evaluate whether children with isolated skull fractures without an underlying brain injury and normal neurological exam require a transfer to a tertiary hospital with pediatric neurosurgery service. Methods A retrospective chart review was performed to review children under five years old presenting to the emergency department of a non-pediatric trauma center with an isolated skull fracture resulting from head trauma without intracerebral hemorrhage between 2015 and 2021. The inclusion criteria consisted of children who have isolated skull fractures without underlying injuries and normal neurological examination. We reviewed these patients' injury characteristics, disposition, and clinical outcomes. The t-test and chi-square were used for evaluating the groups and evaluating the transfer to a dedicated trauma care facility. Results We identified 26 children who had isolated skull fractures with no underlying brain injury and normal neurological examination. The two most common mechanisms of injury were falls (64%) and motor vehicle collisions (MVC) (11%). The median age of patients was six months old. The location of the skull fractures was as follows: parietal (46%), occipital (19%), temporal (15%), frontal (7.7%), occipital + parietal (7.7%), and parietal + frontal (3.8%). Four fractures were depressed (15%) and the remainder were non-displaced. Eleven children with skull fractures (42%) were transferred to a designated pediatric trauma center and the remaining 58% were hospitalized for observation and monitored at the primary hospital. None of the children with skull fractures required intubation or other advanced interventions. Conclusion In this relatively limited sample, approximately one-third of the children with isolated skull fractures without brain injury were managed successfully in a non-tertiary care center. However, none of them required surgical intervention. Thus, we propose that patients akin to those in this study can be observed at a local hospital without being transferred to a pediatric trauma center.

RAD51 Is Implicated in DNA Damage, Chemoresistance and Immune Dysregulation in Solid Tumors.

BACKGROUND: In normal cells, homologous recombination (HR) is tightly regulated and plays an important role in the maintenance of genomic integrity and stability through precise repair of DNA damage. RAD51 is a recombinase that mediates homologous base pairing and strand exchange during DNA repair by HR. Our previous data in multiple myeloma and esophageal adenocarcinoma (EAC) show that dysregulated HR mediates genomic instability. Purpose of this study was to investigate role of HR in genomic instability, chemoresistance and immune dysregulation in solid tumors including colon and breast cancers. METHODS: The GEO dataset were used to investigate correlation of RAD51 expression with patient survival and expression of various immune markers in EAC, breast and colorectal cancers. RAD51 was inhibited in cancer cell lines using shRNAs and a small molecule inhibitor. HR activity was evaluated using a plasmid-based assay, DNA breaks assessed by evaluating expression of γ-H2AX (a marker of DNA breaks) and p-RPA32 (a marker of DNA end resection) using Western blotting. Genomic instability was monitored by investigating micronuclei (a marker of genomic instability). Impact of RAD51 inhibitor and/or a DNA-damaging agent was assessed on viability and apoptosis in EAC, breast and colon cancer cell lines in vitro and in a subcutaneous tumor model of EAC. Impact of RAD51 inhibitor on expression profile was monitored by RNA sequencing. RESULTS: Elevated RAD51 expression correlated with poor survival of EAC, breast and colon cancer patients. RAD51 knockdown in cancer cell lines inhibited DNA end resection and strand exchange activity (key steps in the initiation of HR) as well as spontaneous DNA breaks, whereas its overexpression increased DNA breaks and genomic instability. Treatment of EAC, colon and breast cancer cell lines with a small molecule inhibitor of RAD51 inhibited DNA breaking agent-induced DNA breaks and genomic instability. RAD51 inhibitor potentiated cytotoxicity of DNA breaking agent in all cancer cell types tested in vitro as well as in a subcutaneous model of EAC. Evaluation by RNA sequencing demonstrated that DNA repair and cell cycle related pathways were induced by DNA breaking agent whereas their induction either prevented or reversed by RAD51 inhibitor. In addition, immune-related pathways such as PD-1 and Interferon Signaling were also induced by DNA breaking agent whereas their induction prevented by RAD51 inhibitor. Consistent with these observations, elevated RAD51 expression also correlated with that of genes involved in inflammation and other immune surveillance. CONCLUSIONS: Elevated expression of RAD51 and associated HR activity is involved in spontaneous and DNA damaging agent-induced DNA breaks and genomic instability thus contributing to chemoresistance, immune dysregulation and poor prognosis in cancer. Therefore, inhibitors of RAD51 have great potential as therapeutic agents for EAC, colon, breast and probably other solid tumors.

Colchicine anti-inflammatory therapy for non-intensive care unit hospitalized COVID-19 patients: results from a pilot open-label, randomized controlled clinical trial.

Systemic inflammation is a hallmark of severe coronavirus disease-19 (COVID-19). Anti-inflammatory therapy is considered crucial to modulate the hyperinflammatory response (cytokine storm) in hospitalized COVID-19 patients. There is currently no specific, conclusively proven, cost-efficient, and worldwide available anti-inflammatory therapy available to treat COVID-19 patients with cytokine storm. The present study aimed to investigate the treatment benefit of oral colchicine for hospitalized COVID-19 patients with suspected cytokine storm. Colchicine is an approved drug and possesses multiple anti-inflammatory mechanisms. This was a pilot, open-label randomized controlled clinical trial comparing standard of care (SOC) plus oral colchicine (colchicine arm) vs. SOC alone (control arm) in non-ICU hospitalized COVID-19 patients with suspected cytokine storm. Colchicine treatment was initiated within first 48 hours of admission delivered at 1.5 mg loading dose, followed by 0.5 mg b.i.d. for next 6 days and 0.5 mg q.d. for the second week. A total of 96 patients were randomly allocated to the colchicine (n=48) and control groups (n=48). Both colchicine and control group patients experienced similar clinical outcomes by day 14 of hospitalization. Treatment outcome by day 14 in colchicine vs control arm: recovered and discharged alive: 36 (75.0%) vs. 37 (77.1%), remain admitted after 14-days: 4 (8.3%) vs. 5 (10.4%), ICU transferred: 4 (8.3%) vs. 3 (6.3%), and mortality: 4 (8.3%) vs. 3 (6.3%). The speed of improvement of COVID-19 acute symptoms including shortness of breath, fever, cough, the need of supplementary oxygen, and oxygen saturation level, was almost identical in the two groups. Length of hospitalization was on average 1.5 day shorter in the colchicine group. There was no evidence for a difference between the two groups in the follow-up serum levels of inflammatory biomarkers including C-reactive protein (CRP), D-dimer, lactate dehydrogenase (LDH), ferritin, interleukin-6 (IL-6), high-sensitivity troponin T (hs-TnT) and N-terminal pro b-type natriuretic peptide (NT pro-BNP). According to the results of our study, oral colchicine does not appear to show clinical benefits in non-ICU hospitalized COVID-19 patients with suspected cytokine storm. It is possible that the anti-inflammatory pathways of colchicine are not crucially involved in the pathogenesis of COVID-19.

Superior Vena Cava Syndrome: A Palliative Approach to Treatment.

Superior vena cava (SVC) syndrome is an oncologic emergency of venous congestion due to impaired venous flow through the SVC to the right atrium, leading to potential hemodynamic instability. We report a case of a 78-year-old female patient with a non-symptomatic lung nodule that exhibited rapid growth from its discovery to an enlarging tumor impinging the SVC in less than one month. The short time span from computed tomography (CT) image of the tumor to oncologic emergency required our team to act quickly to identify the source of the tumor and halt its progression, utilizing a multidisciplinary team approach while dealing with a patient that executed their right of autonomy to refusal of care, thus focusing on management with palliative goals since SVC syndrome has a life expectancy of six months post-diagnosis.

Together Till the End: Two Cases of Withdrawal of Life Support.

It's important to consider patients' wishes regarding treatment options, especially near the end of life to allow patients to die with dignity. Worldwide variability exists regarding the palliative extubation decision, which is performed to relieve suffering by the termination of mechanical ventilation and withdrawal of the breathing tube, consequently avoiding the prolongation of death. It is only performed when it is consistent with patients' values and prognosis. This variability is even more prominent in low-income and developing countries. We are presenting a case report of two patients, a husband and a wife, who underwent palliative extubation and withdrawal of life support on the same day.

Multisystem Inflammatory Syndrome in Children.

Multisystem inflammatory syndrome in children (MIS-C) is an uncommon but emerging syndrome related to SARS-CoV-2 infection. While the presentation of MIS-C is generally delayed after exposure to the virus that causes coronavirus 2019, both MIS-C and Kawasaki disease (KD) share similar clinical features. Multisystem inflammatory syndrome in children poses a diagnostic and therapeutic challenge given the lack of definitive diagnostic tests and a paucity of evidence regarding treatment modalities. We review the clinical presentation, diagnostic evaluations, and management of MIS-C and compare its clinical features to those of KD.

Effects of copper nanoparticles on performance, muscle and bone characteristics and serum metabolites in broilers.

Three hundred and twenty day old Hubbard broilers were randomly allocated to four treatments (8 replicates, 10 birds/pen) and were raised under standard management conditions. Birds in the first group served as control and were fed a corn based diet, while birds in the remaining three groups i.e.; A, B and C were fed with a basal diet supplemented with copper nanoparticles (CuNP) at 5, 10 and 15 mg /kg of diet respectively for 35 days. Supplementation of CuNP linearly increased (P≤0.05) body weight (BW), average daily weight gain (ADWG) and feed intake (FI) in broilers. Uric acid, glucose levels in blood and feed conversion ratio (FCR) reduced linearly (P≤0.05) with CuNP supplementation in diet. Supplementation of CuNP in the diet also linearly increased (P≤0.05) tibia weight, length, diameter, weight/length index (W/L) and Tibiotarsal index (TT index). Inclusion of CuNP in broilers diet linearly increased the measured parameters of muscle i.e.; pH, fiber diameter, fiber cross-sectional area, fascicle diameter, fascicle cross-sectional area (P≤0.05). Concentration of copper, iron, calcium and phosphorous in blood also increased line-arly (P ≤ 0.05) with CuNP supplementation. Overall, CuNP positively affected the growth performance, histological characteristics of muscles, bone strength and serum metabolites in broilers.

Fungal conversion of chicken-feather waste into biofortified compost.

Poultry industry is amongst highly developed industries of Pakistan, fulfilling the protein demand of rapidly increasing population. On the other hand, the untreated poultry waste is causing several health and environmental problems. The current study was designed to check the potential of keratinolytic fungal species for the conversion of chicken-feather waste into biofortified compost. For the purpose, three fungal species were isolated from soil samples. These strains were pure cultured and then characterized phenotypically and genotypically. BLAST searches of 18S rDNA nucleotide sequence of the fungal isolates revealed that the two fungal isolates belonged to genus Aspergillus and one belonged to genus Chrysosporium. Optimum temperature for Aspergillus flavus, Aspergillus niger and Chrysosporium queenslandicum was 29, 26 and 25 oC, respectively. A. flavus showed maximum (53%) feather degradation, A. niger degraded feather waste up to 37%, while C. queenslandicum showed 21% keratinolytic activity on chicken feathers at their respective temperature optima. The degradation potential of these fungal species showed their ability to form compost that has agro-industrial importance.