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ObjectiveLysine-Specific Demethylase-1 (LSD1) is overexpressed in breast cancer cells and facilitate mesenchymal properties which may contribute to therapeutic resistance and cancer progression. The purpose of this study was to investigate the safety of combination, nab-paclitaxel and phenelzine, an irreversible LSD1 inhibitor in patients with metastatic breast cancer (mBC).MethodsEligible patients with mBC were treated with nab-paclitaxel (100mg/m2) weekly for 3 weeks with one week break in a 28-day cycle. Dose escalation of phenelzine followed the Cumulative Cohort Design and phenelzine treatment commenced from day 2 of first cycle. Eleven patients were screened, and eligible patients were enrolled in cohorts with the dose of phenelzine ranging from 45mg to 90mg.ResultsThe Optimum Biological Dose was established at 60mg of phenelzine daily in combination with nab-paclitaxel and considered as the recommended phase 2 dose. Most (95%) of adverse events were grade 1 or 2 with two grade 3 events being diarrhea and neutropenia at 45mg and 60mg phenelzine respectively, with no unexpected toxicity/deaths. Commonly reported toxicities were fatigue (n=4,50%), dizziness (n=6,75%), neutropenia (n=3,37.5%), peripheral neuropathy (n=3,37.5%), diarrhea (n=2,25%), and hallucination (n=2,25%). After a median follow up of 113 weeks, all patients showed disease progression on trial with 4 patients being alive at the time of data cut off, including one patient with triple negative breast cancer. Median progression-free survival was 34 weeks. Significant inhibition of LSD1 and suppression of mesenchymal markers in circulating tumor cells were noted.ConclusionPhenelzine in combination with nab-paclitaxel was well tolerated, without any unexpected toxicities in patients with mBC and demonstrated evidence of antitumor activity. For the first time, this proof-of-concept study showed in-vivo inhibition of LSD1 suppressed mesenchymal markers, which are known to facilitate generation of cancer stem cells with metastatic potential. Clinical Trial Registration:ClinicalTrials.Gov NCT03505528, UTN of U1111-1197-5518.

Lysine specific demethylase 1 (LSD1) is a key epigenetic eraser enzyme implicated in cancer metastases and recurrence. Nuclear LSD1 phosphorylated at serine 111 (nLSD1p) has been shown to be critical for the development of breast cancer stem cells. Here we show that circulating tumor cells isolated from immunotherapy-resistant metastatic melanoma patients express higher levels of nLSD1p compared to responders, which is associated with co-expression of stem-like, mesenchymal genes. Targeting nLSD1p with selective nLSD1 inhibitors better inhibits the stem-like mesenchymal signature than traditional FAD-specific LSD1 catalytic inhibitors such as GSK2879552. We also demonstrate that nLSD1p is enriched in PD-1+CD8+ T cells from resistant melanoma patients and 4T1 immunotherapy-resistant mice. Targeting the LSD1p nuclear axis induces IFN-γ/TNF-α-expressing CD8+ T cell infiltration into the tumors of 4T1 immunotherapy-resistant mice, which is further augmented by combined immunotherapy. Underpinning these observations, nLSD1p is regulated by the key T cell exhaustion transcription factor EOMES in dysfunctional CD8+ T cells. EOMES co-exists with nLSD1p in PD-1+CD8+ T cells in resistant patients, and nLSD1p regulates EOMES nuclear dynamics via demethylation/acetylation switching of critical EOMES residues. Using novel antibodies to target these post-translational modifications, we show that EOMES demethylation/acetylation is reciprocally expressed in resistant and responder patients. Overall, we show for the first time that dual inhibition of metastatic cancer cells and re-invigoration of the immune system requires LSD1 inhibitors that target the nLSD1p axis.

Immune literacy--the ability to hear, learn, read, write, explain, and discuss immunological content with varied audiences--has become critically important in recent years. Yet, with its complex terminology and discipline-specific concepts, educating individuals about the immune system and its role in health and disease may seem daunting. Here, we reflect on how to demystify the discipline and increase its accessibility for a broader audience. To address this, a working group of immunology educators from diverse institutions associated with the research coordination network, ImmunoReach, convened virtually. As a result of these discussions, we request a call to action for a system-level change and present a set of practical recommendations that novice and experienced educators from diverse institutions, professional societies, and policymakers may adopt to foster immune literacy in their classrooms and communities.

BackgroundIntravesical Bacillus Calmette–Guérin (BCG) remains the standard adjuvant treatment for non-muscle invasive bladder cancer (NMIBC) following transurethral resection; however, BCG failure and related toxicities are common.ObjectivesTo compare the efficacy and toxicity of intravesical BCG and gemcitabine in the treatment of NMIBC.MethodsRetrospective data were collected in the region of Canberra, Australia from January 2010 to December 2015. The survival cutoff was December 2016. Primary end point was disease-free survival (DFS) and secondary end point was toxicity. After optimal transurethral resection all patients received weekly intravesical BCG or gemcitabine for 6 weeks and maintenance treatment according to their risk. The recurrence was defined as histology proven tumor recurrence (any grade), or appearance of carcinoma in situ.ResultsOne hundred and three patients were evaluable, 52 treated with BCG and 51 with gemcitabine with a median age of 77 and 78, and were mostly male. Approximately half of each received maintenance therapy. The groups were well balanced, apart from some difference in cancer risk groups. Twenty-one percent in the BCG group and 29% in the gemcitabine group had received prior BCG. Median follow up was 15.0 months. Median DFS was 19.6 months for BCG, whereas median DFS was not reached with gemcitabine. There was a trend toward improved DFS with gemcitabine in multivariate analysis, HR: 0.49 (95% CI: 0.22–1.06, p = 0.07). Adverse events were significantly less frequent with gemcitabine (7 versus 44%, p ≤ 0.05). There were four cases of systemic BCG infection.ConclusionIntravesical gemcitabine was associated with a trend toward better DFS with significantly lower toxicity when compared with BCG. Intravesical BCG remains the standard first-line adjuvant therapy; however, intravesical gemcitabine could be a reasonable alternative in cases where BCG is contraindicated and for patients who are intolerant or refractory to BCG. A prospective phase 3 trial is needed to confirm the benefits of gemcitabine over BCG.

DUSP6 is a dual-specificity phosphatase (DUSP) involved in breast cancer progression, recurrence, and metastasis. DUSP6 is predominantly cytoplasmic in HER2+ primary breast cancer cells, but the expression and subcellular localization of DUSPs, especially DUSP6, in HER2-positive circulating tumor cells (CTCs) is unknown. Here we used the DEPArray system to identify and isolate CTCs from metastatic triple negative breast cancer (TNBC) patients and performed single-cell NanoString analysis to quantify cancer pathway gene expression in HER2-positive and HER2-negative CTC populations. All TNBC patients contained HER2-positive CTCs. HER2-positive CTCs were associated with increased ERK1/ERK2 expression, which are direct DUSP6 targets. DUSP6 protein expression was predominantly nuclear in breast CTCs and the brain metastases but not pleura or lung metastases of TNBC patients. Therefore, nuclear DUSP6 may play a role in the association with cancer spreading in TNBC patients, including brain metastasis.

Extra-nodal natural killer/T-cell lymphoma-nasal type (ENKTCL-NT) represents a rare clinical entity that often masquerades as benign lesions during its early stages, resulting in delayed diagnosis and appropriate treatment. The optimal management of this condition hinges on immunohistochemistry, disease stage, and risk stratification based on five critical prognostic factors, which encompass stage, age, performance status, lactate dehydrogenase levels, and primary tumor invasion. Radiation therapy is the cornerstone for achieving locoregional control in localized and advanced disease stages. In this case report, we present the case of a 49-year-old male, who sought medical attention at our center due to uncontrolled epistaxis following an interventional procedure for a suspected nasal polyp. Subsequent examination of histopathology slides and blocks, along with immunohistochemistry, confirmed the diagnosis of ENKTCL-NT, with residual disease evident on positron emission tomography-computed tomography. Following deliberation by the multidisciplinary tumor board, the decision was made to administer radical radiation therapy alone, with a total dose of 5940 cGy (centigray) delivered in 30 fractions employing intensity-modulated radiotherapy techniques, considering the patient's cardiac comorbidities.

Abstract The current study focuses on examining the characteristics of biofuel obtained from the pyrolysis of Madhuca longifolia residues, since the selected forest residue was primarily motivated by its greater volatile matter content. The study used several analytical techniques to describe pyrolysis oil, char, and gas obtained from slow pyrolysis process conducted between 350 and 600 °C in a fixed-bed reactor. Initially, the effect of process temperature on product distribution was assessed to motivate maximum pyrolysis oil yield and found to be 44.2 wt% at pyrolysis temperature of 475 °C, while the yields of char and gas were 22.1 wt% and 33.7 wt%, respectively. In order to determine the suitability of the feedstock, the Madhuca longifolia residues were analyzed by TGA and FT-IR, which revealed that the feedstock could be a feasible option as an energy source. The characterization of pyrolysis oil, char, and gas has been done through various analytical methods like FT-IR, GC-MS, and gas chromatography. The physicochemical characteristics of the pyrolysis oil sample were examined, and the results showed that the oil is a viscous liquid with a lower heating value than conventional diesel. The FT-IR and GC-MS analysis of pyrolysis oil revealed the presence of increased levels of oxygenated chemicals, acids, and phenol derivatives. The findings of the FT-IR analysis of char indicated the existence of aromatic and aliphatic hydrocarbons. The increased carbon content in the char indicated the possibility of using solid fuel. Gas chromatography was used to examine the chemical structure of the pyrolysis gas, and the results showed the existence of combustible elements.

Although introns are typically tens to thousands of nucleotides, there are notable exceptions. In flies as well as humans, a small number of genes contain introns that are more than 1000 times larger than typical introns, exceeding hundreds of kilobases (kb) to megabases (Mb). It remains unknown why gigantic introns exist and how cells overcome the challenges associated with their transcription and RNA processing. The Drosophila Y chromosome contains some of the largest genes identified to date: multiple genes exceed 4Mb, with introns accounting for over 99% of the gene span. Here we demonstrate that co-transcriptional splicing of these gigantic Y-linked genes is important to ensure successful transcription: perturbation of splicing led to the attenuation of transcription, leading to a failure to produce mature mRNA. Cytologically, defective splicing of the Y-linked gigantic genes resulted in disorganization of transcripts within the nucleus suggestive of entanglement of transcripts, likely resulting from unspliced long RNAs. We propose that co-transcriptional splicing maintains the length of nascent transcripts of gigantic genes under a critical threshold, preventing their entanglement and ensuring proper gene expression. Our study reveals a novel biological significance of co-transcriptional splicing.

OBJECTIVES/GOALS: 48,000,000 people in the U.S. have hearing loss, negatively impacting quality of life and work. Unveiling axon guidance for auditory type II spiral ganglia neurons (SGNs) will aid development of new therapies. I study the role of Eph/Ephrin and planar cell polarity (PCP) signaling during type II SGN turning and outer hair cell (OHC) innervation. METHODS/STUDY POPULATION: This quantitative study was conducted on Efna3 and Vangl2 null mice possessing Neurog1CreERT2 and R26RtdTomato mutations. Spontaneous Cre activity within the Neurogenin1CreERT2 line causes recombination and expression of fluorescent Rosa26 Reporter (R26R)tdTomatoin a restricted number of SGNs, including type IIs. Together, these lines permit SGN sparse labeling. Immunostaining and confocal imaging were used to analyze dsRed in Efna3 and Vangl2 mice and quantify type II SGN turning. In combination, Imaris 3D renderings were used to quantify type II SGN turning, branching, navigation features and temporal effects of EPHRIN-A3-Fc on type IIs via cochlear cultures (a gain-of-function manipulation). For both sexes, 5-6 cochleae per genotype were analyzed and compared by t-test to wildtype (WT) controls. RESULTS/ANTICIPATED RESULTS: Efna3 nulls showed a small rise in type II SGNs incorrectly turning toward the apex at an error rate of 16.0% compared to WTs (n=6; p=0.05). P0 Efna3 nulls had reduced branch number compared to WTs, 4.1 and 7.2, respectively (n=129; p=

Paramedics are exposed to many infectious diseases in their professional activities, leading to a high risk of transmitting infectious diseases to patients in out-of-hospital settings, possibly leading to health care associated infections in hospitals and the community. The COVID-19 pandemic highlighted the importance of infection prevention and control in health care and the role of paramedics in infection control is considered even more critical. Despite this, in many countries such as Australia, research into infection prevention and control research has mainly been focused on in-hospital health care professionals with limited out-of-hospital studies. This scoping review was based upon Preferred Reporting Items for Systematic Reviews and Meta-Analysis guidelines. Literature on knowledge and awareness of infection prevention and control in paramedics in Australia and other countries was evaluated. Based upon selection criteria applied, six papers were identified for inclusion in this review. In many studies, infection prevention and control was identified as being important, however compliance with hand hygiene practices was low and most studies highlighted the need for more education and training on infectious disease for paramedics. Current evidence suggests that paramedics have poor compliance with recommended IPC practices. The profession needs to improve IPC education, training, and culture. • Overall, paramedics have poor compliance with infection prevention and control practices (IPC). • Compared with other health professions the paramedicine profession is behind on IPC practices. • There is urgent need for further research into IPC practices in paramedicine. [ABSTRACT FROM AUTHOR]

Introduction/background COVID-19 vaccination uptake rates and responses by Canadian respiratory therapists (RTs) were investigated along with factors that may be shown to play a role in vaccination hesitancy. Methods An anonymous survey using SurveyMonkey® on vaccination uptake rates, responses and attitudes was made available to student RTs, graduate RTs and registered RTs in Canada from July to October of 2021. Pearson's chi-square tests were performed to evaluate association between vaccination status and the other categorical parameters evaluated. Results A total of 1013 surveys (8.0% of target population) were completed fully and included in the data analysis. Of the surveyed RT population, 90.42% received their vaccination as soon as it was made available compared to Canada's Ministry of Health's published rate at the time of 86.27% for all Canadian healthcare workers. There was a significant (p = 0.013) association between early vaccination and age and a significant (p = 0.036) association between vaccination status and a participant's response on whether or not they have a family member or know someone who has had COVID-19. There was also a significant (p \< 0.001) association between vaccination status and attitudes towards trusting science to develop safe, effective, new vaccines and trusting the Ministry of Health to ensure that vaccines are safe. There was no significant association between vaccination status and gender, province/territory of residency/work, level of education and level of involvement with COVID-19 patients. Conclusion The results suggest that RT groups across Canada had higher early vaccination uptake rates than general healthcare worker groups and that age, relationship to people with COVID-19 and trust in science played a significant role in their vaccination uptake rates.

Apicomplexan parasites are leading causes of human and livestock diseases such as malaria and toxoplasmosis, yet most of their genes remain uncharacterized. Here, we present the first genome-wide genetic screen of an apicomplexan. We adapted CRISPR/Cas9 to assess the contribution of each gene from the parasite Toxoplasma gondii during infection of human fibroblasts. Our analysis defines ∼200 previously uncharacterized, fitness-conferring genes unique to the phylum, from which 16 were investigated, revealing essential functions during infection of human cells. Secondary screens identify as an invasion factor the claudin-like apicomplexan microneme protein (CLAMP), which resembles mammalian tight-junction proteins and localizes to secretory organelles, making it critical to the initiation of infection. CLAMP is present throughout sequenced apicomplexan genomes and is essential during the asexual stages of the malaria parasite Plasmodium falciparum. These results provide broad-based functional information on T. gondii genes and will facilitate future approaches to expand the horizon of antiparasitic interventions.

Glioblastoma (GBM) harbors subpopulations of therapy-resistant tumor-initiating cells (TICs) that are self-renewing and multipotent. To understand the regulation of the TIC state, we performed an image-based screen for genes regulating GBM TIC maintenance and identified ZFHX4, a 397 kDa transcription factor. ZFHX4 is required to maintain TIC-associated and normal human neural precursor cell phenotypes in vitro, suggesting that ZFHX4 regulates differentiation, and its suppression increases glioma-free survival in intracranial xenografts. ZFHX4 interacts with CHD4, a core member of the nucleosome remodeling and deacetylase (NuRD) complex. ZFHX4 and CHD4 bind to overlapping sets of genomic loci and control similar gene expression programs. Using expression data derived from GBM patients, we found that ZFHX4 significantly affects CHD4-mediated gene expression perturbations, which defines ZFHX4 as a master regulator of CHD4. These observations define ZFHX4 as a regulatory factor that links the chromatin-remodeling NuRD complex and the GBM TIC state.

Plant diseases are a major threat to agriculture, particularly impacting the yield and quality of tomato crops. Early detection and accurate classification of these diseases are essential for effective management and mitigation. Traditional methods of disease detection are often labor-intensive and time-consuming. Although individual convolutional neural networks (CNNs) have shown promise in automated plant disease detection, their accuracy and robustness can be limited when used in isolation. This study proposes an ensemble learning approach that combines three state-of-the-art CNN architectures: AlexNet, ResNet50, and VGG16. A comprehensive dataset of tomato leaf images, categorized into bacterial, viral, fungal diseases, and healthy leaves, was used. Images were preprocessed and augmented to improve model generalization. Each model was trained separately, and their outputs were integrated using a weighted averaging mechanism to form the ensemble model. The weights for each model were optimized based on validation performance. The ensemble model significantly improved classification accuracy compared to individual models. The combined approach achieved an overall accuracy of 97.5%, with precision, recall, and F1-score exceeding 95% for all disease categories. Specifically, the accuracy for detecting bacterial diseases was 96.8%, viral diseases 97.2%, and fungal diseases 97.9%. The ensemble method demonstrated superior robustness and reliability in classifying diverse disease symptoms [ABSTRACT FROM AUTHOR]

It has long been held as scientific fact that soon after birth, cardiomyocytes cease dividing, thus explaining the limited restoration of cardiac function after a heart attack. Recent demonstrations of cardiac myocyte differentiation observed in vitro or after in vivo transplantation of adult stem cells from blood, fat, skeletal muscle, or heart have challenged this view. Analysis of these studies has been complicated by the large disparity in the magnitude of effects seen by different groups and obscured by the recently appreciated process of in vivo stem-cell fusion. We now show a novel population of nonsatellite cells in adult murine skeletal muscle that progress under standard primary cell-culture conditions to autonomously beating cardiomyocytes. Their differentiation into beating cardiomyocytes is characterized here by video microscopy, confocal-detected calcium transients, electron microscopy, immunofluorescent cardiac-specific markers, and single-cell patch recordings of cardiac action potentials. Within 2 d after tail-vein injection of these marked cells into a mouse model of acute infarction, the marked cells are visible in the heart. By 6 d they begin to differentiate without fusing to recipient cardiac cells. Three months later, the tagged cells are visible as striated heart muscle restricted to the region of the cardiac infarct.

Telehealth refers to the use of telecommunication devices and other forms of technology to provide services outside of the traditional in‐person health care delivery system. Growth in the use of telehealth creates new challenges and opportunities for implementation in clinical practice. The American Academy of Physical Medicine and Rehabilitation (AAPM&R) assembled an expert group to develop a white paper to examine telehealth innovation in Physical Medicine and Rehabilitation (PM&R). The resultant white paper summarizes how telehealth is best used in the field of PM&R while highlighting current knowledge deficits and technological limitations. The report identifies new and transformative opportunities for PM&R to advance translational research related to telehealth and enhance patient care. [ABSTRACT FROM AUTHOR]

Objective: To summarize the recommendations on the interventional management of subacute and chronic nonradicular low back pain (LBP) from the 21 quality‐appraised clinical practice guidelines (CPGs) identified in a previously published paper: "Quality of Clinical Practice Guidelines on Interventional Management of Low Back Pain: A Systematic Review." By disseminating this information, we aimed to facilitate the implementation of these recommendations into clinical practice. Type: Systematic Review Literature Survey: Electronic bibliographic databases, guideline databases, and gray literature were searched from January 2016 to January 2020 to identify CPGs that met study criteria. Methodology: A total of 21 CPGs were quality appraised and interventional management recommendations were extracted and organized into several treatment categories including epidural steroid injections, radiofrequency procedures (RF), facet injections, sacroiliac injections, and prolotherapy. Within each treatment category, the recommendations were organized based on two factors: quality of CPG and strength of recommendation. Synthesis: Overall, there was no consistency in recommendations for or against any interventional procedure, even when accounting for the quality of the CPG. In all of the CPGs reviewed, the most common strength of recommendation was weakly for. The second, third, and fourth most common strength of recommendations were inconclusive, weakly against, and strongly against, respectively, and the least common was strongly for. The treatment categories with the greatest number of recommendations were RF procedures (most common strength of recommendation was weakly for) and facet procedures. Among the high‐quality CPGs, the most common strength of recommendation was inconclusive. Conclusions: Most of the interventional management recommendations for management of nonradicular LBP in the 21 CPGs appraised in this review were either weakly for, weakly against, or inconclusive, with several recommendations within each treatment category contradicting each other. Appraisal of Guidelines for Research & Evaluation Instrument quality appraisals of CPGs on interventional management of LBP were of unclear utility in guiding clinical implementation. [ABSTRACT FROM AUTHOR]

Objective: To appraise the quality of low back pain (LBP) clinical practice guidelines (CPG) that include interventional management recommendations and to associate their quality with characteristics including publication year and creating organization. Type Systematic Review. Literature Survey: LBP (subacute or chronic) CPGs in English (symptom based, governmental or professional society created, January 1990‐May 2020) were found using MEDLINE, EMBASE, CINAHL, Ortho Guidelines, CPG Infobase, ECRI, Guidelines International Network, NICE, and SIGN. Methodology: In this third order systematic review, search results were deduplicated, title and abstract screened by two independent reviewers, and full texts reviewed by four reviewers. Discrepancies were resolved by a third reviewer. Resulted CPGs were appraised using the Appraisal of Guidelines for Research and Evaluation (AGREE II) Tool by four appraisers each. Association of their quality with creating organization, geographical region, and year of creation was calculated. Synthesis: Seven hundred fourteen screened documents resulted in 21 final CPGs. On appraisal, average overall CPG quality was 5.2 (range 2.5‐6.75). Domain 5 (applicability) had the lowest average (44%) and domain 4 (clarity of presentation) had the highest average score (82%). For overall recommendation, 16 received "yes" or "yes with modifications," six received unanimous "yes" and two unanimous "no" votes. The interrater agreement of domain scoring was excellent (0.8‐1.0; p <.001). There was no association found between quality of CPG and (1) year of publication (R2 = 0.0006), (2) whether the CPG was updated or new (p =.17), and (3) region of publication (p =.37). Conclusions: The majority of the 21 CPGs identified in this systematic review were of high quality, but overall quality and recommendation ratings were variable. The quality of appraised CPGs showed no association with their characteristics. Some domains such as "applicability" scored uniformly lower, revealing opportunity for improvement in future CPG development. LBP CPGs should be scrutinized before adopting their recommendations. [ABSTRACT FROM AUTHOR]