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Sugarcane, the world's most harvested crop by tonnage, has shaped global history, trade and geopolitics, and is currently responsible for 80% of sugar production worldwide1. While traditional sugarcane breeding methods have effectively generated cultivars adapted to new environments and pathogens, sugar yield improvements have recently plateaued2. The cessation of yield gains may be due to limited genetic diversity within breeding populations, long breeding cycles and the complexity of its genome, the latter preventing breeders from taking advantage of the recent explosion of whole-genome sequencing that has benefited many other crops. Thus, modern sugarcane hybrids are the last remaining major crop without a reference-quality genome. Here we take a major step towards advancing sugarcane biotechnology by generating a polyploid reference genome for R570, a typical modern cultivar derived from interspecific hybridization between the domesticated species (Saccharum officinarum) and the wild species (Saccharum spontaneum). In contrast to the existing single haplotype ('monoploid') representation of R570, our 8.7 billion base assembly contains a complete representation of unique DNA sequences across the approximately 12 chromosome copies in this polyploid genome. Using this highly contiguous genome assembly, we filled a previously unsized gap within an R570 physical genetic map to describe the likely causal genes underlying the single-copy Bru1 brown rust resistance locus. This polyploid genome assembly with fine-grain descriptions of genome architecture and molecular targets for biotechnology will help accelerate molecular and transgenic breeding and adaptation of sugarcane to future environmental conditions.

The direction of individual B8 solar neutrinos has been reconstructed using the SNO+ liquid scintillator detector. Prompt, directional Cherenkov light was separated from the slower, isotropic scintillation light using time information, and a maximum likelihood method was used to reconstruct the direction of individual scattered electrons. A clear directional signal was observed, correlated with the solar angle. The observation was aided by a period of low primary fluor concentration that resulted in a slower scintillator decay time. This is the first time that event-by-event direction reconstruction in high light-yield liquid scintillator has been demonstrated in a large-scale detector.

Chao-Bang Ma,1,* Cheng-Yang Zhang,1,* Cai-Li Gou,1 Zeng-Hui Liang,1 Jing-Xian Zhang,2 Fei Xing,1,3 Jing-Jing Yuan,1,3 Xin Wei,1,3 Ya-Bing Zhang,1 Zhong-Yu Wang1,3 1Department of Anesthesiology, Pain and Perioperative Medicine, the First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan Province, People’s Republic of China; 3Henan Province International Joint Laboratory of Pain, Cognition and Emotion, Zhengzhou, Henan Province, People’s Republic of China; 2The Surgical Department of the First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan Province, People’s Republic of China*These authors contributed equally to this workCorrespondence: Zhong-Yu Wang, Department of Anesthesiology, Pain and Perioperative Medicine, the First Affiliated Hospital of Zhengzhou University, No. 1 Jianshe East Road, Erqi District, Zhengzhou City, Henan Province, People’s Republic of China, Tel +86-13937165514, Email wzy781217@163.comPurpose: Postoperative delirium (POD) is a prevalent and severe complication in elderly patients undergoing major surgery, associated with increased morbidity and mortality. This randomized controlled trial aimed to investigate the effects of low-dose esketamine on the incidence of POD in elderly patients underwent total hip or knee arthroplasty.Patients and Methods: Two hundred and sixty elderly participants were randomly assigned to either the esketamine group (Group E) (0.20mg/kg loading, 0.125mg/kg/h infusion, 0.5 mg/kg for postoperative analgesia) or the placebo group (Group P) (received normal saline). The primary outcome was the incidence of POD, with secondary outcomes including delirium subtypes, duration, intraoperative analgesic consumption, operative and anesthesia times, hemodynamic changes, postoperative pain scores, sleep quality, and common postoperative adverse events.Results: There was no significant difference in the incidence of POD between Group E (8.5%) and Group P (10.8%). No significant differences were observed for the time of delirium onset, duration of delirium, and delirium subtype between the two groups. Patients in the esketamine group had more stable hemodynamic profile after induction and reduced the pain score of motion on the first two days postoperatively but increased the incidence of postoperative dizziness.Conclusion: The repeated infusion of low-dose esketamine did not reduce the incidence of POD during the initial three postoperative days in elderly patients following total hip or knee arthroplasty.Keywords: esketamine, delirium, aged, arthroplasty

Caroline B Kurtz,1 Mona S Calvo2 1Synlogic Inc., Cambridge, MA, USA; 2Icahn School of Medicine at Mount Sinai, Manhattan, NY, USACorrespondence: Caroline B Kurtz, Email carolinebkurtz@gmail.comAbstract: Dietary supplements represent a growing segment of the healthcare industry as consumers seek alternative methods to manage health concerns. Probiotics are dietary supplements that are broadly used to aid digestion and immune and metabolic health. Patients with disorders restricting the intake of particular nutrients must balance adequate nutrition with limitations in the intake of natural food ingredients. Synthetic biology has emerged in the last decade, allowing scientists to specifically engineer probiotic bacteria to perform functions in the GI tract, such as the elimination or reduction of specific nutrients from ingested food sources. Engineered probiotic bacteria have the potential to provide health benefits and improve the quality of life of patients with genetic or acquired metabolic disorders. One way to make them available to consumers is to develop specialized products as dietary supplements. Developers of such innovative products must follow new FDA guidance and work closely with FDA in consideration of the design, development, manufacturing, and labeling of engineered probiotics as dietary supplements.Keywords: probiotic dietary supplement, FDA, new dietary ingredient

Ishak Ahmed Abdi,1 Ahmed Elmi Abdi,1 Mohamed Omar Hassan,1 Said Abdirahman Ahmed,1 Mohamud Mire Waberi,1 Ahmed Shafie Aden,1 Osman Farah Dahir,1 Mohamed Ahmed Mohamud,2 Irshad Ibrahim Ali,3 Shuayb Moallim Ali Jama,4 Abdirahman Mohamed Hassan Dirie,5 Can Baba Arın6 1Department of Cardiology, Mogadishu Somali Türkiye Training and Research Hospital, Mogadishu, Somalia; 2Internal Medicine Resident, Kenyatta National Hospital, Nairobi, Kenya; 3Anesthesia and Critical Care Unit, Mogadishu Somali Türkiye Training and Research Hospital, Mogadishu, Somalia; 4Department of Radiology, Mogadishu Somali Türkiye Training and Research Hospital, Mogadishu, Somalia; 5Respiratory Medicine Department, Mogadishu Somali Türkiye Training and Research Hospital, Mogadishu, Somalia; 6Department of Cardiology, Health Science University, Dr. Siyami Ersek Thoracic and Cardiovascular Surgery Center, Training and Research Hospital, İstanbul, TurkeyCorrespondence: Ishak Ahmed Abdi, Department of Cardiology, Mogadishu Somali Türkiye Training and Research Hospital, Second Street, Black Sea, Hodan District, Mogadishu, Somalia, Tel +252618439596, Email isakabdi70@gmail.comIntroduction: Pulmonary embolism (PE) is a critical condition that can present with diverse and often non-specific electrocardiographic findings. The Aslanger pattern, identified in 2020, is an uncommon electrocardiography (ECG) finding typically associated with acute non-ST-segment elevation myocardial infarction (NSTEMI). Although this pattern is usually linked to myocardial infarction with significant coronary stenosis, we report a rare case where the Aslanger pattern was observed in a patient with acute pulmonary embolism.Case Presentation: A 52-year-old male with Parkinson’s disease presented with sudden chest pain, shortness of breath, sweating, and fatigue. Initial ECG showed Aslanger pattern, suggesting acute coronary syndrome. Despite elevated troponin I (1624 ng/mL) and normal coronary angiography, further investigation revealed right ventricular dilation and massive pulmonary embolism confirmed by CT angiography. The patient was treated with thrombolytics and anticoagulation, stabilizing over five days and discharged on rivaroxaban.Conclusion: The Aslanger pattern usually linked to myocardial infarction can also occur in acute pulmonary embolism. This case underscores the need for careful differential diagnosis and timely treatment to prevent complications associated with delayed care.Keywords: pulmonary embolism, Aslanger pattern, electrocardiogram

Polaritons, i.e., hybrid quasi-particles of light and matter resonances, have been extensively investigated due to their potential to enhance light-matter interactions. Although polaritonic applications thrive in the mid-infrared range, their extension to the terahertz (THz) range remains limited. Here, we present paratellurite (α-TeO2) nanowires, a versatile material acting as a platform for different types of phonon polaritons. Utilizing synchrotron infrared nanospectroscopy from 10 to 24 THz, we uncover the polaritonic properties of α-TeO2 nanowires, showcasing their dual functionality as both a Fabry-Pérot cavity and a waveguide for surface phonon polaritons. Furthermore, near-field measurements with a free-electron laser as a THz source reveal a localized optical contrast down to 5.5 THz, an indication of hyperbolic bands. Our findings complement the repertoire of polaritonic materials, with significant implications for advancing THz technologies.

A repeat expansion mutation in the C9orf72 gene is the leading known genetic cause of FTD and ALS. The C9orf72-ALS/FTD field has been plagued by a lack of reliable tools to monitor this genomic locus and its RNA and protein products. We have validated assays that quantify C9orf72 pathobiology at the DNA, RNA and protein levels using knock-out human iPSC lines as controls. Here we show that single-molecule sequencing can accurately measure the repeat expansion and faithfully report on changes to the C9orf72 locus in what has been a traditionally hard to sequence genomic region. This is of particular value to sizing and phasing the repeat expansion and determining changes to the gene locus after gene editing. We developed ddPCR assays to quantify two major C9orf72 transcript variants, which we validated by selective excision of their distinct transcriptional start sites. Using validated knock-out human iPSC lines, we validated 4 commercially available antibodies (of 9 tested) that were specific for C9orf72 protein quantification by Western blot, but none were specific for immunocytochemistry. We tested 15 combinations of antibodies against dipeptide repeat proteins (DPRs) across 66 concentrations using MSD immunoassay, and found two (against poly-GA and poly-GP) that yielded a 1.5-fold or greater signal increase in patient iPSC-motor neurons compared to knock-out control, and validated them in human postmortem and transgenic mouse brain tissue. Our validated DNA, RNA and protein assays are applicable to discovery research as well as clinical trials.

Abstract High-brightness particle beams generated by advanced accelerator concepts have the potential to become an essential part of future accelerator technology. In particular, high-gradient accelerators can generate and rapidly accelerate particle beams to relativistic energies. The rapid acceleration and strong confining fields can minimize irreversible detrimental effects to the beam brightness that occur at low beam energies, such as emittance growth or pulse elongation caused by space charge forces. Due to the high accelerating gradients, these novel accelerators are also significantly more compact than conventional technology. Advanced accelerators can be extremely variable and are capable of generating particle beams with vastly different properties using the same driver and setup with only modest changes to the interaction parameters. So far, efforts have mainly been focused on the generation of electron beams, but there are concepts to extend the sources to generate spin-polarized electron beams or positron beams. The beam parameters of these particle sources are largely determined by the injection and subsequent acceleration processes. Although, over the last decade there has been significant progress, the sources are still lacking a sufficiently high 6-dimensional (D) phase-space density that includes small transverse emittance, small energy spread and high charge, and operation at high repetition rate. This is required for future particle colliders with a sufficiently high luminosity or for more near-term applications, such as enabling the operation of free-electron lasers (FELs) in the X-ray regime. Major research and development efforts are required to address these limitations in order to realize these approaches for a front-end injector for a future collider or next-generation light sources. In particular, this includes methods to control and manipulate the phase-space and spin degrees-of-freedom of ultrashort plasma-based electron bunches with high accuracy, and methods that increase efficiency and repetition rate. These efforts also include the development of high-resolution diagnostics, such as full 6D phase-space measurements, beam polarimetry and high-fidelity simulation tools. A further increase in beam luminosity can be achieve through emittance damping. Emittance cooling via the emission of synchrotron radiation using current technology requires kilometer-scale damping rings. For future colliders, the damping rings might be replaced by a substantially more compact plasma-based approach. Here, plasma wigglers with significantly stronger magnetic fields are used instead of permanent-magnet based wigglers to achieve similar damping performance but over a two orders of magnitude reduced length.

It is widely accepted that the next lepton collider beyond a Higgs factory would require center-of-mass energy of the order of up to 15 TeV. Since, given reasonable space and cost restrictions, conventional accelerator technology reaches its limits near this energy, high-gradient advanced acceleration concepts are attractive. Advanced and novel accelerators (ANAs) are leading candidates due to their ability to produce acceleration gradients on the order of 1-100 GV/m, leading to compact acceleration facilities. However, intermediate energy facilities (IEF) are required to test the critical technology elements on the way towards multi-TeV-class collliders. Here a science case for a 20-100 GeV center-of-mass energy ANA-based lepton collider that can be a candidate for an intermediate energy facility is presented. The IEF can provide numerous opportunities for high energy physics studies including precision Quantum Chromodynamics and Beyond the Standard Model physics measurements, investigation of charged particle interactions with extreme electromagnetic fields, and exploring muon and proton beam acceleration. Possible applications of this collider include the studies of γγ and electron beam-fixed target/beamdump collider designs. Thus, the goal of the proposed IEF is to both carry out particle physics measurements in the 20-100 GeV ranges as well as to serve as an ANA demonstrator facility.

The Building Seismic Safety Council (BSSC) recently completed a multi-year effort to rewrite Chapter 16 of the ASCE/SEI 7-10 Standard (2010), which governs ground motion selection and modification for new building projects in the United States. This paper provides an overview of new recommended provisions regarding the use of spectral matching of ground motions. Example analyses of three buildings are shown to illustrate the reasoning behind the spectral matching language, and to demonstrate the relative differences in response estimates obtained from scaled versus spectrally matched motions. For these examples, the demands placed on the buildings by spectrally matched motions developed using the recommended provisions were comparable to the demands produced by amplitude scaled motions. The consistent demands produced by the procedures suggest that evaluations of a building design’s acceptability are comparable whether scaled or spectrally matched motions are used. The revised provisions should significantly advance the practice of ground motion selection and modification in the United States and elsewhere, by aligning practice with research outcomes.

Kush S Patel,1 Ronald J Anderson,1,* Carolyn B Becker,1,* William C Taylor,1,* Anne F Liu,1,* Anubodh S Varshney,2,* Nadaa B Ali,1,* Barbara J Nath,3,* Stephen R Pelletier,4 Helen M Shields,1 Nora Y Osman1 1Department of Medicine, Brigham and Women’s Hospital, Boston, MA, USA; 2Division of Cardiovascular Medicine, Department of Medicine, Stanford University, Palo Alto, CA, USA; 3Department of Medicine, Massachusetts General Hospital, Boston, MA, USA; 4Office of Medical Education, Harvard Medical School, Boston, MA, USA*These authors contributed equally to this workCorrespondence: Helen M Shields, Department of Medicine, Brigham and Women’s Hospital, 75 Francis St, Boston, MA, 02115, USA, Tel +1 617 678 6077, Fax +1 617 525 8740, Email hmshields@bwh.harvard.eduPurpose: Coaching is a well-described means of providing real-time, actionable feedback to learners. We aimed to determine whether dual coaching from faculty physicians and real inpatients led to an improvement in history-taking skills of clerkship medical students.Patients and Methods: Expert faculty physicians (on Zoom) directly observed 13 clerkship medical students as they obtained a history from 26 real, hospitalized inpatients (in person), after which students received immediate feedback from both the physician and the patient. De-identified audio-video recordings of all interviews were scored by independent judges using a previously validated clinical rating tool to assess for improvement in history-taking skills between the two interviews. Finally, all participants completed a survey with Likert scale questions and free-text prompts.Results: Students’ history-taking skills – specifically in the domains of communication, medical knowledge and professional conduct – on the validated rating tool, as evaluated by the independent judges, did not significantly improve between their first and second patient interviews. However, students rated the dual coaching as overwhelmingly positive (average score of 1.43, with 1 being Excellent and 5 being Poor), with many appreciating the specificity and timeliness of the feedback. Patients also rated the experience very highly (average score of 1.23, with 1 being Excellent and 5 being Poor), noting that they gained new insights into medical training.Conclusion: Students value receiving immediate and specific feedback and real patients enjoy participating in the feedback process. Dual physician-patient coaching is a unique way to incorporate more direct observation into undergraduate medical education curricula.Keywords: direct observation, immediate feedback, medical student education, real patients, virtual teaching

Anwar E Ahmed,1 Callista B Martin,2 Bassam Dahman,3 Gregory T Chesnut,4,5 Sean Q Kern4,5 1Department of Preventive Medicine and Biostatistics, Uniformed Services University of the Health Sciences, Bethesda, MD, USA; 2School of Medicine, Uniformed Services University of the Health Sciences, Bethesda, MD, USA; 3Department of Health Behavior and Policy, Virginia Commonwealth University, Richmond, VA, USA; 4Department of Surgery, Uniformed Services University of the Health Sciences, Bethesda, MD, USA; 5Center for Prostate Disease Research, Murtha Cancer Center, Uniformed Services University of the Health Sciences, Bethesda, MD, USACorrespondence: Anwar E Ahmed, Uniformed Services University of the Health Sciences, 4301 Jones Bridge Road A-1039D, Bethesda, MD, 20814, USA, Tel +1 301-295-9756, Email anwar.ahmed@usuhs.eduBackground: Research suggests inconsistent evidence regarding the association between general obesity and prostate cancer among men in the United States. This study aimed to examine whether the association between general obesity and prostate cancer is influenced by abdominal obesity and ethnic groups.Methods: The study utilized data from the National Health and Nutrition Examination Survey (NHANES). The analysis was restricted to non-Hispanic men (10,683 White and 6,020 Black). Obesity was defined as body mass index (BMI) ≥ 30 and abdominal obesity as waist circumference (WC) ≥ 102 cm.Results: No significant difference was identified in the overall prevalence of prostate cancer between obese and non-obese (2.14% vs 2.25%, P = 0.678). When both obesity measures were combined, the general and abdominal obesity category was associated with a significant increase in the odds of prostate cancer in Black men [odds ratio (OR) = 1.49, 95% confidence interval (CI) (1.09, 2.04)], but not in White men [OR = 1.29, 95% CI (0.91, 1.82)]. In both Black [OR = 2.46, 95% CI (1.48, 4.06)] and White men [OR = 1.60, 95% CI (1.16, 2.21)], abdominal obesity was associated with significant increase in the odds of prostate cancer.Conclusion: The association between general obesity and prevalence of prostate cancer depends on abdominal obesity and ethnic groups. Our study utilized a nationally representative survey and emphasized the potential of combined effect of general and abdominal obesity as a modifiable factor to decrease racial disparity in prostate cancer screening and poor outcomes.Keywords: prostate cancer, waist circumference, obesity, Black men, racial disparity

Ya-Ya Bai,1 Rui Tian,1 Yan Qian,2 Qiao Zhang,1 Chong-Bo Zhao,1 Yong-Gang Yan,1 Li Zhang,3 Shi-Jun Yue,4 Yu-Ping Tang1 1Key Laboratory of Shaanxi Administration of Traditional Chinese Medicine for TCM Compatibility, and State Key Laboratory of Research & Development of Characteristic Qin Medicine Resources (Cultivation), and Shaanxi Collaborative Innovation Center of Chinese Medicinal Resources Industrialization, and Shaanxi Traditional Chinese Medicine Processing Technology Heritage Base, Shaanxi University of Chinese Medicine, Xi’an, 712046, People’s Republic of China; 2Suzhou Institute for Drug Control, Suzhou, Jiangsu Province, 215000, People’s Republic of China; 3Hanlin College, Nanjing University of Chinese Medicine, Taizhou, Jiangsu Province, 225300, People’s Republic of China; 4International Joint Research Center on Resource Utilization and Quality Evaluation of Traditional Chinese Medicine of Hebei Province, Hebei University of Chinese Medicine, Shijiazhuang, 050200, People’s Republic of ChinaCorrespondence: Qiao Zhang, Shaanxi University of Chinese Medicine, Shiji Avenue, Xi Xian New District, Xi’an, 712046, People’s Republic of China, Tel +86-15895973680, Fax: +86-02938185061, Email 18700081184@163.comBackground: Long-term use of rhubarb (RH) commonly leads to diarrhea, which can be alleviated by steaming with wine. However, the specific mechanism by which wine steaming alleviates RH-induced diarrhea remains unknown.Objective: This study aims to reveal the underlying mechanisms of wine steaming in alleviating RH-induced diarrhea by examining small intestinal flora and serum metabolomics.Methods: Major anthraquinone and anthrone components were detected using ultra-performance liquid chromatography-mass spectrometry (UPLC-MS). Eighty-four ICR mice were randomly divided into control, RH, and RH steamed with wine (PRH) groups and were administered RH and PRH (1, 4, and 8 g/kg, i.g). for 14 consecutive days. Histopathological analysis was performed using hematoxylin-eosin staining. Levels of inflammatory factors and tight junction proteins, zonula occludens-1 (ZO-1) and occludin, in the small intestine were measured. The small intestine content was analyzed using 16S rRNA sequencing, and UPLC-MS was used to analyze endogenous metabolites.Results: Levels of major anthraquinone and anthrone components decreased in PRH. Both RH and PRH groups showed varying degrees of loose stools and increased fecal water rates; the RH group exhibited more severe effects. Compared with the control group, RH caused small intestine injuries, increased levels of inflammatory cytokines, downregulated the expression of ZO-1 and occludin, and induced gut microbiota (GM) imbalance. The relative abundance of Lactobacillus decreased, while the relative abundance of Shigella and Streptococcus increased. However, PRH had a milder impact than RH. The glycerophospholipid metabolic pathway was involved in this effect. The levels of inflammatory cytokines and potential metabolites (sn-glycero-3-phosphoethanolamine) were positively correlated with Streptococcus infection, while the levels of ZO-1 and occludin were negatively correlated with Streptococcus infection. GM imbalance and abnormal glycerophospholipid metabolism contributed to impaired intestinal barrier function and inflammatory factor release, which may underlie RH-induced diarrhea, though PRH had a weaker effect.Conclusion: PRH alleviated RH-induced diarrhea by recovering GM balance, reducing ZO-1 and occludin expression, and decreasing the release of inflammatory factors. This mechanism may be linked to the reduced anthraquinone content. This study is the first to explore the mechanism of wine steaming in alleviating RH-induced diarrhea through small intestinal flora and serum metabolomics. It provides data to support the broader clinical use of RH and its safer application. Keywords: gut microbiota, metabolomics, wine steaming, diarrhea, rhubarb

ABSTRACT Aims/Introduction Two Hong Kong Chinese non‐laboratory‐based prediabetes/diabetes mellitus (pre‐DM/DM) risk models were developed using logistic regression (LR) and machine learning, respectively. We aimed to evaluate the models' validity in case finding of pre‐DM/DM in a Chinese primary care (PC) population. We also evaluated the validity of a risk‐scoring algorithm derived from the LR model. Materials and Methods This was a cross‐sectional external validation study on Chinese adults, without a prior DM diagnosis, who were recruited from public/private PC clinics in Hong Kong. A total of 1,237 participants completed a questionnaire on the models' predictors. Of that, 919 underwent blood glucose testing. The primary outcome was the models' and the algorithm's sensitivity in finding pre‐DM/DM cases. The secondary outcomes were the models' and the algorithm's specificity, positive/negative predictive values, discrimination and calibration. Results The models' sensitivity were 0.70 (machine learning) and 0.72 (LR). Both showed good external discrimination (area under the receiver operating characteristic curve: machine learning 0.744, LR 0.739). The risks estimated by the models were lower than the observed incidence, indicating poor calibration. Both models were more effective among participants with lower pretest probabilities; that is, age 18–44 years. The algorithm's sensitivity was 0.77 at the cut‐off score of ≥16 out of 41. Conclusion This study showed the validity of the models and the algorithm for finding pre‐DM/DM cases in a Chinese PC population in Hong Kong. They can facilitate more cost‐effective identification of high‐risk individuals for blood testing to diagnose pre‐DM/DM in PC. Further studies should recalibrate the models for more precise risk estimation in PC populations.

Allison F O’Neill,1 Raul C Ribeiro,2 Emilia M Pinto,3 Michael R Clay,4 Gerard P Zambetti,3 Brent A Orr,3 Christopher B Weldon,5 Carlos Rodriguez-Galindo2,6 1Department of Pediatric Oncology, Dana-Farber/Boston Children’s Cancer and Blood Disorders Center, Boston, MA, USA; 2Department of Oncology, St. Jude Children’s Research Hospital, Memphis, TN, USA; 3Department of Pathology, St. Jude Children’s Research Hospital, Memphis, TN, USA; 4Department of Pathology, Children’s Hospital Colorado, Denver, CO, USA; 5Department of Surgery, Boston Children’s Hospital, Boston, MA, USA; 6Department of Global Pediatric Medicine, St. Jude Children’s Research Hospital, Memphis, TN, USACorrespondence: Allison F O’Neill, Department of Pediatric Oncology, Dana-Farber Cancer Institute, 450 Brookline Avenue – DA3111, Boston, MA, 02215, USA, Email Allison_oneill@dfci.harvard.eduAbstract: Adrenocortical tumors (ACTs) are infrequent neoplasms in children and adolescents and are typically associated with clinical symptoms reflective of androgen overproduction. Pediatric ACTs typically occur in the context of a germline TP53 mutation, can be cured when diagnosed at an early stage, but are difficult to treat when advanced or associated with concurrent TP53 and ATRX alterations. Recent work has demonstrated DNA methylation patterns suggestive of prognostic significance. While current treatment standards rely heavily upon surgical resection, chemotherapy, and hormonal modulation, small cohort studies suggest promise for multi-tyrosine kinases targeting anti-angiogenic pathways or immunomodulatory therapies. Future work will focus on novel risk stratification algorithms and combination therapies intended to mitigate toxicity for patients with perceived low-risk disease while intensifying therapy or accelerating discoveries aimed at improving survival for patients with difficult-to-treat disease.Keywords: adrenocortical, pediatric, management, discovery, carcinoma, tumors

Muhammad B Qureshi,1 Jose O Garcia,1 Jaxon Quillen,2 Carolyn Mead-Harvey,2 Christina Wentz,3 Cherie B Nau,4 Muriel Schornack,4 Keith Baratz,4 Sanjay V Patel,4 Joanne Shen5 1Mayo Clinic Alix School of Medicine, Scottsdale, AZ, USA; 2Department of Quantitative Health Sciences, Mayo Clinic, Scottsdale, AZ, USA; 3Illinois College of Optometry, Chicago, IL, USA; 4Department of Ophthalmology, Mayo Clinic, Rochester, MN, USA; 5Department of Ophthalmology, Mayo Clinic, Scottsdale, AZ, USACorrespondence: Joanne Shen, Department of Ophthalmology, Mayo Clinic, 13400 E Shea Blvd, Scottsdale, AZ, USA, 85259, Tel +1 480-301-8085, Fax +1 480-301-7326, Email shen.joanne@mayo.eduPurpose: To compare baseline characteristics and treatment of chronic ocular graft-versus-host disease (oGVHD) patients in two treatment locations.Patients and Methods: Patients diagnosed with definite chronic oGVHD between September 1, 2014 and September 20, 2021 at two locations were identified. IRB-approved retrospective chart review was conducted for the following data: demographic information, ocular surface disease index (OSDI), corneal fluorescein staining (CFS), and treatment(s) used. Differences by site were assessed using Pearson’s Chi-Square tests and two-sample t-tests; differences by time were assessed using paired t-tests.Results: At baseline, Clinic 1 (C1) patients had a worse mean OSDI score (47.8 vs 36.3, p = 0.011) and CFS in both OD (1.3 vs 0.8, p = 0.005) and OS (1.3 vs 0.6, p < 0.001) compared to Clinic 2 (C2). Comparing baseline to endpoint, C1 patients experienced an improvement in OSDI (− 17.26, p < 0.001), CFS OD (− 0.50, p < 0.001), and CFS OS (− 0.51, p < 0.001) at C1. Change in OSDI, CFS OD, or CFS OS was not statistically significant at C2. Despite similar follow-up length, C1 demonstrated more clinic visits (10.4 vs 3.4, p < 0.001) and more treatment trials (4.9 vs 2.4, p < 0.001) compared to C2. Punctal plugs (85.5% vs 61.2%, p = 0.002), punctal cautery (69.7% vs 28.6%, p < 0.001), topical steroids (72.4% vs 22.4%, p < 0.001), and autologous serum tears (AST) (52.6% vs 8.2%, p < 0.001) were used more frequently at C1 than at C2.Conclusion: oGVHD patients at C1 experienced significant improvement in OSDI and corneal fluorescein staining and compared to patients at C2, had more frequent follow-up and use of punctal plugs, punctal cautery, topical steroids, and AST.Keywords: graft-versus-host disease, GVHD, chronic GVHD, dry eye disease, ocular GVHD, keratoconjunctivitis sicca, KCS

Stig Holm Ovesen,1,2 Søren Helbo Skaarup,3 Rasmus Aagaard,4 Hans Kirkegaard,1 Bo Løfgren,1,5 Michael Dan Arvig,6– 8 Bo Martin Bibby,9 Stefan Posth,10 Christian B Laursen,11,12 Jesper Weile1,2 1Research Center for Emergency Medicine, Department of Clinical Medicine, Aarhus University Hospital and Aarhus University, Aarhus, Denmark; 2Emergency Department, Horsens Regional Hospital, Horsens, Denmark; 3Department of Respiratory Diseases and Allergy, Aarhus University Hospital, Aarhus, Denmark; 4Department of Anesthesiology, Aarhus University Hospital, Aarhus, Denmark; 5Department of Internal Medicine, Randers Regional Hospital, Randers, Denmark; 6Emergency Department, Slagelse Hospital, Slagelse, Denmark; 7Department of Clinical Medicine, University of Copenhagen, Copenhagen, Denmark; 8Department of Regional Health Research, University of Southern Denmark, Odense, Denmark; 9Department of Biostatistics, Aarhus University, Aarhus, Denmark; 10Emergency Department, Odense University Hospital, Odense, Denmark; 11Department of Respiratory Medicine, Odense University Hospital, Odense, Denmark; 12Odense Respiratory Research Unit (ODIN), Department of Clinical Research, University of Southern Denmark, Odense, DenmarkCorrespondence: Stig Holm Ovesen, Research Center for Emergency Medicine, Aarhus University Hospital, Palle Juul-Jensens Boulevard 99, J103, Aarhus, 8200, Denmark, Email stigholm@clin.au.dkPurpose: Point-of-care ultrasound (POCUS) helps emergency department (ED) physicians make prompt and appropriate decisions, but the optimal diagnostic integration and potential clinical benefits remain unclear. We describe the protocol and statistical analysis plan for a randomized controlled trial. The objective is to determine the effect of a POCUS-driven diagnostic pathway in adult dyspneic ED patients on the proportion of patients having a hospital stay of less than 24 hours when compared to the standard diagnostic pathway.Patients and Methods: This is a multicenter, randomized, investigator-initiated, open-labeled, pragmatic, controlled trial. Adult ED patients with chief complaint dyspnea are eligible. Patients are randomized (1:1) to the POCUS-driven diagnostic pathway or standard diagnostic pathway, with 337 patients in each group. The primary outcome is the proportion of patients having a hospital stay (from ED arrival to hospital discharge) of less than 24 hours. Key secondary outcomes include hospital length-of-stay, 72-hour revisits, and 30-day hospital-free days.Conclusion: Sparse evidence exists for any clinical benefit from a POCUS-integrated diagnostic pathway. The results from this trial will help clarify the promising signals for POCUS to influence patient care among ED patients with dyspnea.Keywords: focused lung ultrasound, focused cardiac ultrasound, shortness-of-breath, diagnostic effectiveness

The Manchester–Oxford Foot Questionnaire is a validated English 16-item patient-reported outcome measure for evaluating foot and ankle function. This study aimed to translate a Cantonese Chinese version of the questionnaire and evaluate its validity and reliability among patients with foot and ankle disorders. The translation and cross-cultural adaptation were conducted by forward translation, synthesis of translated versions, backward translation, review, and pre-testing by a panel of healthcare professionals. The test-retest reliability was conducted within 2–4 weeks. Internal consistency and construct validity were evaluated using the Visual Analogue Scale and Short-Form 36. 135 participants were recruited to complete the validation process. The overall reliability coefficient constituted 0.86 (intraclass correlation coefficient = 0.72–0.93). A strong, statistically significant correlation was found between the bodily pain subscale of Short-Form 36 and the pain subscale of the Cantonese Chinese-Manchester-Oxford Foot Questionnaire ( r = −0.77, p

Background The management of pulmonary nodules plays a critical role in early detection of lung cancer. Computed tomography (CT) has led to a stage-shift towards early-stage lung cancer, but regional differences in survival rates have been reported in Denmark. This study aimed to evaluate whether variations in nodule management among Danish health regions contributed to these differences.Material and Methods The Danish Health Data Authority and Danish Lung Cancer Registry provided data on CT usage and lung cancer stage distribution, respectively. Auditing of lung cancer stage IA patient referrals and nodule management of stage IV lung cancer patients was conducted in seven Danish lung cancer investigation centers, covering four of the five Danish health regions. CT scans were performed up to 2 years before the patients’ diagnosis from 2019 to 2021.Results CT usage has increased steadily in Denmark over the past decade, with a simultaneous increase in the proportion of early-stage lung cancers, particularly stage IA. However, one Danish health region, Region Zealand, exhibited lower rates of early-stage lung cancer and overall survival despite a CT usage roughly similar to that of the other health regions. The audit did not find significant differences in pulmonary nodule management or a higher number of missed nodules by radiologists in this region compared to others.Conclusion This study suggests that a high CT scan volume alone is not sufficient for the early detection of lung cancer. Factors beyond hospital management practices, such as patient-related delays in socioeconomically disadvantaged areas, may contribute to regional differences in survival rates. This has implications for future strategies for reducing these differences.

The LUX-ZEPLIN experiment recently reported limits on WIMP-nucleus interactions from its initial science run, down to 9.2×10-48 cm2 for the spin-independent interaction of a 36 GeV/c2 WIMP at 90% confidence level. In this paper, we present a comprehensive analysis of the backgrounds important for this result and for other upcoming physics analyses, including neutrinoless double-beta decay searches and effective field theory interpretations of LUX-ZEPLIN data. We confirm that the in-situ determinations of bulk and fixed radioactive backgrounds are consistent with expectations from the ex-situ assays. The observed background rate after WIMP search criteria were applied was (6.3±0.5)×10-5 events/keVee/kg/day in the low-energy region, approximately 60 times lower than the equivalent rate reported by the LUX experiment.

Preserving the quality of a positron beam in a plasma-based accelerator, where a wakefield suitable for positron transport and acceleration is generated by means of an electron filament, is challenging. This is due to the nature of the wakefields, characterized by focusing fields that vary nonlinearly in the transverse direction, and by accelerating fields that are non-uniform. These fields also change slice-by-slice along the beam. Maintaining a high beam quality is pivotal for application of positron beams in a plasma-based collider. In this paper, we show that an initial background plasma temperature can help mitigate the positron beam quality degradation in plasma-based accelerators that rely on electron filaments. We show that temperature effects broaden the electron filament and smooth radially both the non-linear transverse and the non-uniform longitudinal wakefields. Using warm plasmas opens up new possibilities to improve beam quality in several plasma-based positron acceleration concepts.

In parallel to the growth of paediatric gastroenterology sub-speciality, gastrointestinal disorders requiring endoscopy for diagnosis or therapy have shown a rising incidence globally. With the development of fibre-optic endoscope, gastrointestinal endoscopy has become a revolutionary diagnostic as well as therapeutic tool. Generally, endoscopy has shown superiority in terms of diagnostic yield over earlier methods of diagnosing common as well as rare gastrointestinal disorders. This review sought to review the recent advances in gastrointestinal endoscopy modalities which have led to the explosion in its use for diagnosis and therapy of different gastrointestinal disorders in children.

Background: Sub- Sahara Africa including Nigeria has the second largest global burden of chronic carriers of hepatitis B virus (HBV) infection after Asia. Mother-to-child transmission (MTCT) of BV is the most common route of transmission in high endemic areas .MTCT of hepatitis B virus infection continues to occur despite the interventions of hepatitis B vaccinations and immunoglobulins in settings where it is practiced. Infants most at risk are those whose mothers have high HBV DNA viral loads and produce the protein HBeAg. Various Nigerian studies have reported high HBV infection rates as well as BeAg positivity among pregnant women. These HBV infections usually occur intrapartum and rarely inutero. Mothers with HBeAg positivity known to be associated with higher HBV DNA viral loads have been linked with higher chances of MTCT as HBeAg is the only structural HBV protein that can cross into the placental circulation. In the absence of post exposure prophylaxis about 40 percent of infants delivered by HBV infected mothers could develop HBV infections, and about 25% of them may come down with chronic hepatitis and resulting possible complications including liver cirrhosis and hepatocellular cancer later in life. The prevention of transmission of retroviral infection from motherto- child has been a success story of the 21st century and such feat could be replicated for HBV infection. The standard PMTCT of HBV currently will comprise: timely prenatal screening, starting anti-viral therapy for pregnant women with HBeAg positivity and high viral load, infant postexposure prophylaxis and followup of infants of HBsAg positive mothers. There is no co-ordinated PMTCT of HBV programme in place in our setting despite the huge burden of the disease in Nigeria. Hence the need therefore to develop a home grown PMTCT programme of HBV to help tackle the burden of the disease in our country. An evidence based review of current best practice guidelines for the prevention of mother-to-child transmission of HBV for use in low and medium resource income settings with hepatitis B hyperendemicity will be quiet apt in this ircumstance. This document therefore will be useful as a quick guide to Paediatricians, Obstetricians, Family Physicians, General Practice Doctors and other allied health workers charged with the care of pregnant mothers and their young children. Methods: Relevant literatures published in English language or translated into English were searched manually and electronically in PUBMED and SCOPUS for the period between 1990 and 2016 on the subject. Keywords searched included: epidemiology of HBV infection, MTCT of HBV, and its preventive strategies including prenatal screening, antiviralagents in pregnancy, infant post exposure prophylaxis and follow-up of infected children. Results: Over 35 scholarly articles on HBV epidemiology, MTCT, and preventive measures as well as follow –up models were retrieved and analyzed. Conclusion: Universal screening of all pregnant women for HBV infection is the most effective strategy for the prevention of MTCT of HBV, as effective preventive measures could be applied starting from pregnancy to delivery while infants of HBsAg positive mothers should receive timely post exposure prophylaxis and followed up for possible development of chronic hepatitis B infection.

Background: Parental sex education of children is an often overlooked issue in pediatrics, especially in our society where talking about issues concerning sex is regarded as a taboo. Objectives: The objective of this study is to determine the impact of sex education on child sexual abuse among adolescents attending secondary schools in south east Nigeria. Methods: This is a cross-sectional study that was carried out among children in three secondary schools in Enugu and Ebonyi states of Nigeria. Five hundred and six adolescents who met inclusion criteria were consecutively recruited into our prospective study between June and October, 2014.Results: Eighty (80%) of respondents were educated by parents on sex abuse. Of those educated majority was by mother (46.2%) and both parents (45.2%). Most (72.1%) were not informed that family members or family friends can abuse children and 73.8% were not told to inform adults if it happens to them. A lower proportion (37.9%) of those educated by parents at home was abused. Those educated by parents were 1.23 times less likely to be abused than those not educated. There was significant difference in sex abuse between males and females (p=0.014) while there were no significant difference for age (p=0.157) and social class (p=0.233). Conclusion: Children educated by parents on sexual abuse were less likely to be sexually abused than those not educated. There is no link between socioeconomic class and child sexual abuse among adolescents.

Background: Obesity once considered a high income country’s malady is now on the rise in most developing countries particularly in urban settings. Most of these emerging economies have been reported to have different shades of under – nutrition coexisting side by side with over-nutrition. It is pertinent therefore that we determine the factors driving the increase in obesity rates in developing countries as they generally lack the infrastructure to adequately handle the associated complications. Objectives: This communication is aimed at reviewing the burden and risk factors for obesity in children in developing countries, double burden of malnutrition, challenges including medical as well as economic costs and sustainable preventive programmes of obesity in our environment with the hope of sensitizing both the health community and policy makers of this emerging epidemic. Methods: We searched relevant literature on the subject published only in English language or translated into English Language manually and electronically. The Index Medicus, AJOL, Medline, PUBMED, and HINARI were specifically searched for the period between 1980 and 2014 and reviewed. The following key words were applied in the search: Obesity in childhood, its burden and associated risk factors, complications of obesity in childhood, double burden of malnutrition in developing countries, assessment of obesity, childhood challenges of obesity including its direct and indirect costs in developing countries as well as practical preventive models in developing economies. Results: Several relevant studies were identified. The health as well as economic costs of obesity is diverse. Obesity is the major risk factor for a variety of non – communicable diseases including cardiovascular diseases, type 2 diabetes and malignancies in later life. Also obese children have higher risk of orthopaedic problems and psychological disturbances like low -self esteem and bullying. This can also lead to poor social adjustments among our teeming youths who are the bedrock of our future economy. Most of these diseases cause premature deaths in addition to long term morbidities. Many of these obesity associated complications impose substantial burden on the health care system in developing countries with weak health systems, and if allowed unmitigated the implications are that the cost of its care may overwhelm not only the health budget but also affect the provisions of basic social amenities. Conclusions: Preventive programmes have been shown to reduce the burden of obesity in developed countries. Dearth of data on burden of obesity and its associated complications in children and adolescents still a challenge in most developing economies. Efforts should be made to prevent childhood obesity using multi- pronged approach at population level through targeted education, sustainable interventions related to healthy nutritional practices as well as physical activity promotion.

Background: In paediatric practice, weight and height are required for therapeutic and diagnostic interventions. In some circumstances actual anthropometric measurements are not possible and estimates are used. Several formulae are in use for weight and height estimations. The adequacy of these estimates has not been tested for our children. The aim of the current study was to compare the adequacy of formula methods of weight and height estimation with measured values in children. Materials and Methods: This was a comparative observational study. Children who met the inclusion criteria were selected consecutively and studied over a two month period using a semi-structured questionnaire. Weight and height of each child were measured and recorded to the nearest 0.1kg and 0.1cm respectively using standard protocols. Weight and height for age were also estimated using the universally accepted formulae. Data were analyzed using SPSS 19.0. Paired t- test was used to compare the means of actual and estimated weights and heights according to age. The level of significance was set at p

Background: In the sub- Saharan Africa, one in every five children dies before their 5th birthday, mostly from preventable causes. Recent data f rom Nigeria, show infant and under-5 mortality rates of 88 and 143 deaths per 1000 live births respectively. Although significant efforts have been made in the last two decades towards the reduction of childhood mortality, the rate still remains unacceptably high. Objective: The aim of the current study was to evaluate the pattern and causes of paediatric (childhood) mortality at Mile 4 Mission Hospital, Abakaliki. This will help us to know exactly what happens in a small mission hospital with few facilities and limited skilled medical personnel. In addition, this is the very first time this study was undertaken in this hospital Materials and Methods: The records of admissions and deaths of children managed at the Mile 4 Mission Hospital from the 1st January 2009 to 31st December 2009 were retrieved from the Medical Records Department for analysis. Results: Of the 1110 patients admitted within this period, 72 died giving an overall mortality of 6.5%. There were 43 males and 29 females, giving a male: female ratio of 1.5:1. Most of the deaths 58 (80.6%) occurred among children aged younger than 2 years. Malaria was the most common presumed cause of death, followed by gastroenteritis and bronchopneumonia. Twelve and half percent (12.5%) of the children died within 24 hours of admission. The commonest causes of death within 24 hours were severe anaemia, pneumonia and gastroenteritis. The modal months for childhood mortality in this hospital were May and August. Conclusion: Mortality was commonest among children younger than 2 years of age. Malaria, gastroenteritis and pneumonia were the commonest causes of death. A proactive planning taking into account the seasonal variation of these diseases could reduce the childhood mortality in this centre

Claus B Juhl,1– 4 Else Marie Bladbjerg,3,5 Bibi Gram,3 Torben Knudsen,3,6 Mette Munk Lauridsen,3,6 Niels-Peter Brøchner Nygaard,1,3,7 Nina Drøjdahl Ryg,1,2,4 Lars Skadhauge,3,8 Anna-Marie Bloch Münster3,5 1Department of Endocrinology, University Hospital of Southern Denmark, Esbjerg, Denmark; 2Steno Diabetes Center Odense, University Hospital of Southern Denmark, Odense, Denmark; 3University of Southern Denmark, Department of Regional Health Research, Odense, Denmark; 4OPEN, Open Patient Data Explorative, Odense University Hospital, Odense, Denmark; 5Department of Clinical Biochemistry, University Hospital of Southern Denmark, Esbjerg, Denmark; 6Department of Gastroenterology, University Hospital of Southern Denmark, Esbjerg, Denmark; 7Department of Neurology, University Hospital of Southern Denmark, Esbjerg, Denmark; 8Department of Occupational Medicine, University Hospital of Southern Denmark, Esbjerg, DenmarkCorrespondence: Claus B Juhl, Department of Endocrinology, University Hospital Southern Denmark, Esbjerg, DK-6700, Denmark, Tel +45 6086 7172, Email Claus.Bogh.Juhl@rsyd.dkPurpose: The prevalence of obesity continues to rise. People with obesity are at increased risk of several diseases. We tested an algorithm-based screening program for people with a BMI above 30 kg/m2 and present data on the prevalence of previously undiagnosed obesity-related diseases.Patients and Methods: Seven hundred and sixty-nine persons with BMI > 30 kg/m2 and age 18– 60 years were screened for diabetes (assessed by glycosylated hemoglobin and oral glucose tolerance test at HbA1c 43– 48 mmol/mol), sleep apnea (screened by questionnaires and assessed by cardiorespiratory monitoring at indication of sleep disorder), liver steatosis or liver fibrosis (assessed by biochemistry and fibroscan) and arterial hypertension (assessed by both office and 24-hour blood pressure measurement). A reference group of people with a BMI of 18.5– 29.9 kg/m2 was established.Results: Of those referred, 73.0% were women. We identified new diabetes in 4.2%, prediabetes in 9.1%, moderate-to-severe sleep apnea in 25.1%, increased liver fat and increased liver stiffness in 68.1% and 17.4%, respectively, and hypertension or masked hypertension in 19.0%. The prevalence of diseases was much higher among men and increased with BMI. Except for hypertension, we found few participants with undiagnosed disease in the reference group.Conclusion: An algorithm-based screening program is feasible and reveals undiagnosed obesity-related disease in a large proportion of the participants. The disproportional referral pattern calls for a tailored approach aiming to include more men with obesity.Trial Registration: Inclusion of the non-obese group was approved by the Scientific Ethics Committee of The Region of Southern Denmark (project identification number: S-20210091), and the study was reported at clinicaltrials.gov (NCT05176132).Plain Language Summary: The number of people with obesity is going up, and they are at a higher risk for various diseases. We tested a screening program for people referred with a BMI over 30 kg/m2 and presented the prevalence of diseases related to obesity. We screened 769 people aged 18 to 60 years with a BMI over 30 kg/m2 for diabetes (biochemistry and glucose tolerance test), sleep apnea (both questionnaires and home monitoring), liver disease (biochemistry and liver scan) and high blood pressure (office and 24-hour readings). We also tested a reference group of people with BMI 18.5-30 kg/m2. Among those screened, 73.0% were women. We found new cases of diabetes in 4.2%, prediabetes in 9.1%, sleep apnea in 25.1%, increased liver fat in 68.1%, increased liver stiffness in 17.4%, and hypertension or masked hypertension in 19.0%. The diseases were more common in men and increased with both higher BMI and age. Except for hypertension, we found few cases in the reference groups. The screening program uncovered undiagnosed obesity-related diseases in a large group of individuals. The uneven distribution of referrals suggests we need a customized approach to include more men with obesity.Keywords: diabetes, pre-diabetes, sleep apnea, metabolic dysfunction-associated steatotic liver disease, hypertension

Le-Ying Ni,1– 3,&ast; Cheng-Biao Ding,1,2,4,&ast; Ji-Min Deng,5 Zheng-Wei Wu,4,6,7 Yun Zhou1,2 1Department of Rehabilitation Medicine, The Second Affiliated Hospital of Anhui Medical University, Hefei, Anhui, People’s Republic of China; 2Research Center for Translational Medicine, The Second Affiliated Hospital of Anhui Medical University, Hefei, Anhui, 230601, People’s Republic of China; 3Department of Rehabilitation Medicine, Maanshan People’s Hospital, Maanshan, Anhui, People’s Republic of China; 4School of Nuclear Science and Technology, University of Science and Technology of China, Hefei, People’s Republic of China; 5Anhui institute for Food and Drug Control, Hefei, People’s Republic of China; 6CAS Key Laboratory of Geospace Environment, University of Science and Technology of China, Hefei, People’s Republic of China; 7Institute of Advanced Technology, University of Science and Technology of China, Hefei, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Zheng-Wei Wu, University of Science and Technology of China, Hefei, 230026, People’s Republic of China, Tel/Fax +86-551-63606045, Email wuzw@ustc.edu.cn Yun Zhou, The Second Affiliated Hospital of Anhui Medical University, Hefei, Anhui, 230601, People’s Republic of China, Tel +86-0551-65997010, Email zhouyunanhui@sina.comBackground: Rheumatoid arthritis fibroblast-like synovial cells (RA-FLS) have become the core effector cells for the progression of rheumatoid arthritis due to their “tumor-like cell” characteristics, such as being able to break free from growth restrictions caused by contact inhibition, promoting angiogenesis, invading surrounding tissues, and leading to uncontrolled synovial growth. In recent years, cold air plasma (CAP) has been widely recognized for its clear anticancer effect. Inspired by this, this study investigated the inhibitory effect of CAP on the tumor-like biological behavior of RA-FLS through in vitro experiments.Methods: Treatment of RA-FLS with CAP at different time doses (0s, 30s, 60s, 120s). 5-ethynyl-2’-deoxyuridine (EdU) proliferation assay was used to determine the cell viability. Analysis of cell migration and invasion was performed by wound-healing assay, transwell assay and immunofluorescent staining for f-actin, respectively. Flow cytometry technique was used for analysis of cell cycle and determination of reactive oxygen species (ROS). Hoechst staining was used for analysis of cell apoptosis. Protein expression was analyzed by Western blot analysis.Results: Molecular and cellular level mechanisms have revealed that CAP blocks RA-FLS in the G2/M phase by increasing intracellular reactive oxygen species (ROS), leading to increased apoptosis and significantly reduced migration and invasion ability of RA-FLS.Conclusion: Overall, CAP has significant anti proliferative, migratory, and invasive effects on RA-FLS. This study reveals a new targeted treatment strategy for RA.Keywords: cold air plasma, rheumatoid arthritis fibroblast-like synovial cells, proliferation, apoptosis, migration

The SNO+ Collaboration reports the first evidence of reactor antineutrinos in a Cherenkov detector. The nearest nuclear reactors are located 240 km away in Ontario, Canada. This analysis uses events with energies lower than in any previous analysis with a large water Cherenkov detector. Two analytical methods are used to distinguish reactor antineutrinos from background events in 190 days of data and yield consistent evidence for antineutrinos with a combined significance of 3.5σ.

A doença de Castleman (DC), foi relatada desde 1954 por Benjamin Castleman e compreende um grupo heterogêneo de alterações histopatológicas nos linfonodos. É classificada em unicêntrica (UDC) ou multicêntrica (MDC) a qual é dividida em MCD idiopática (iMCD), MCD associada ao (HHV8) (HHV8-MCD) e polineuropatia, organomegalia, endocrinopatia, distúrbio monoclonal de células plasmáticas, MCD associada a alterações cutâneas (POEMS) (POEMS-MCD). Pode ser classificada em variante hialina-vascular (HV), plasmocítica (PC) e mista. Para pacientes com DCU, a ressecção completa da lesão é o padrão-ouro, e a taxa de SG em 5 anos se aproxima de 100%. Anticorpos monoclonais, imunomodulares, corticosteróides e quimioterapia (QT) compõem o arsenal terapêutico para MCD (HIV- e HHV8-). A poliomielite é uma doença infecciosa, causada pelo poliovírus selvagem, um membro da família Picornaviridae, da espécie Enterovirus C., transmitida por contaminação fecal-oral com replicação linfática orofaríngea e GI. Caracteriza-se por paralisia flácida aguda e pode haver sequela motora residual. Com a vacinação global, os casos de poliovírus selvagem diminuíram mais de 99% desde 1988, passando de 350 000 casos em 125 países endémicos, para 2 países endémicos – Paquistão e Afeganistão (em 10/2023, OMS). Em 1994, a Região das Américas da OMS foi certificada como livre da poliomielite. Objetivo: Descrever o caso de um paciente com DC rica em plasmócitos - multicêntrica - HHV8 negativa em paciente HIV negativo pós poliomielite na Amazonia oriental. Caso clínico: Paciente em 2021 notou linfonodomegalia inguinal direita de crescimento progressivo. Apresentou paralisia infantil com sequela motora em dimídio direito e uso de muleta unilateral. Relata que em 2023 houve crescimento do linfonodo e em 27/07/23 realizou USG com multiplos linfonodos acometendo a cadeia ilíaca a direita, a região inguinal e femoral deste lado, os maiores na região inguinofemoral. de 4,5 x 3,1 cm, sugerindo linfonodos patológicos. Em 18/12/2023 realizou exérese de tumoração da região inguinal direita com AP de processo linfoproliferativo nodular com frequentes folículos regressivos e proliferação vascular na zona interfolicular. achados sugestivos de DC, forma hialino-vascular. IHQ: Doença de Castleman, forma rica em plasmócitos, HHV8 negativa. TC de abdome total de 29/12/23: múltiplas linfonodomegalias comprometendo as cadeias: inguinal direita, iliacas externa, interna e comum direitas, assim como ilíacas comum e interna esquerdas, sendo a maior delas na cadeia ilíaca externa direita de 3,8 cm. Padrão de impregnação hipervascular ao meio de contraste, com leve proeminência sinusoidal (linfedema) adjacente e obliteração adiposa periférica, achados que sugerem a possibilidade de doença linfoproliferativa, destacando-se a etiologia de DC. TC de pescoço de 29/12/2023 com proeminência numérica dos linfonodos das cadeias cervicais, de forma difusa bilateral. Optado por QT com R-COP, com resposta após 4 ciclos. Método: Estudo descritivo do tipo estudo de Caso. Os dados foram avaliados e registrados no programa Microsoft Word 2010. Conclusão: Em geral, a DC é uma doença altamente heterogênea. Não encontramos, na literatura médica pesquisada, caso semelhante ao aqui relatado sobre uma correlação direta entre poliovírus e MCD. Considerando que praticamente todos os países estão livres de Poliomielite, destaca-se a potencial exiguidade de outros casos e a importância do relato para a comunidade científica.

Objetivo: Determinar os intervalos de referência (IR) de índices hematológicos derivados de parâmetros do hemograma em indivíduos saudáveis. Material e métodos: Este estudo incluiu participantes da linha de base de Minas Gerais e Rio Grande do Sul do estudo ELSA-Brasil com informações do hemograma. Foram excluídos participantes com proteína C reativa (PCR) > 10 mg/dL; que auto relataram: problemas de saúde nas últimas 12h, uso de medicamento contínuo, saúde regular, ruim, muito ruim, diabetes mellitus, hipertensão arterial, doença cardiovascular, doença hepática, câncer e tabagismo; obesidade (IMC ≥ 30 kg/m2), taxa de filtração glomerular < 60 mL/min e com subtipos leucocitários fora do IR estabelecido pela Pesquisa Nacional de Saúde. Os índices avaliados foram: razões neutrófilos/linfócitos (RNL), plaquetas/linfócitos (RPL), linfócitos/monócitos (RLM), neutrófilos/global leucócitos – neutrófilos (dNLR) (derived NLR), neutrófilos x monócitos/linfócitos (índice de resposta à inflamação sistêmica - SIRI), neutrófilos x monócitos x plaquetas/linfócitos (índice agregado de inflamação sistêmica - AISI), plaquetas x neutrófilos/linfócitos (índice de imunoinflamação sistêmica - SII). O teste de Dixon foi utilizado para verificar outliers e o teste Kolmogorov-Smirnov para avaliar a distribuição. O IR correspondeu ao intervalo entre os percentis 2,5 e 97,5. O teste Mann Whitney verificou diferenças entre subgrupos de sexo e idade e indicar IR específica para faixa etária e sexo pela estatística de Harris/Boyd. Resultados: Após aplicar os critérios de exclusão, a amostra analítica totalizou 759 indivíduos dos 5.176 avaliados. A maioria era homem (54%), com idade entre 35-59 anos (92%). Nenhum parâmetro apresentou relação estatística (p < 0,05) com a idade. Quanto ao sexo, os valores das medianas da RML e SIRI foram estatisticamente maiores nos homens e RPL, RLM e SII nas mulheres. Os IR foram categorizados apenas para os parâmetros que atenderam a pelo menos um critério da abordagem estatística Harris/Boyd e as recomendações do documento C28A3 do Clinical and Laboratory Standards Institute (CLSI). Os IR obtidos foram: RNL [0.84-3.77]; RLM [1.95-7.39]; dNLR [0.65-2.50]; SIRI [0.29-2.19]; AISI [61.72–521.60]; SII [168.17-871.72]; RPL [mulheres: 75.99–236.34; homens: 69.31–206.10]; RML [mulheres: 0.13–0.45; homens 0.15–0.53]. Discussão: Os índices hematológicos derivados do hemograma têm sido estudados em muitos contextos clínicos, como nas doenças cardíacas e neoplásicas, porém ainda não há pontos de cortes ou IR para auxiliar seu uso. Esse estudo é pioneiro, propondo IR para estes índices em uma população brasileira saudável. Limitações incluem amostra restrita a adultos com ≥ 35 anos, da coorte ELSA que não inclui extremos da hierarquia social do país. Conclusão: Devido à qualidade dos dados do ELSA-Brasil foi possível garantir que uma amostra de referência saudável fosse selecionada e estabelecidos os IR para os índices hematológicos derivados do hemograma. Nossos resultados podem ser úteis para consolidar o uso desses índices, contribuindo para sua padronização e validação na prática clínica.

A validação de metodologias e processos deve ser realizada pelo serviço de hemoterapia para demonstrar que seus resultados são confiáveis e atendem aos atributos de qualidade definidos nas normativas que regem o Ciclo do Sangue. A contagem de hemácias, leucócitos e plaquetas é um dos ensaios realizados no controle de qualidade dos hemocomponentes produzidos, e trata-se de metodologia quantitativa que deve assegurar a obtenção de resultados precisos e exatos. Objetivo Validar a contagem manual de hemácias, leucócitos e plaquetas. Material e métodos As contagens foram realizadas em amostras de sangue total, de controle hematológico comercial ou de plasma fresco. A diluição das amostras foi realizada em cloreto de sódio 0,9% para a contagem de hemácias (1:400 – Câmara de Neubauer); em solução de Turk para a contagem de leucócitos (1:100 – Câmara de Neubauer ou 1:10 – Câmara de Nageotte); ou em solução de Oxalato de amônio 1% para a contagem de plaquetas (1:200 – Câmara de Neubauer). Foram determinadas a repetibilidade, precisão intermediária e exatidão, e a contagem manual foi comparada com a contagem automática por meio da Análise de Concordância de Métodos de Bland-Altman. Os critérios de aceitação para repetibilidade e precisão intermediária foi o coeficiente de variação (CV) ≤ 10% e erro relativo (ER) de ± 15% para os ensaios de exatidão. Resultados Nos ensaios de repetibilidade, o CV foi < 10% para as contagens de hemácias, leucócitos e plaquetas realizadas por todos os técnicos do laboratório. O CV nos ensaios de precisão intermediária foi de 2,01% na contagem de hemácias; de 7,64% na contagem de plaquetas; 6,41% e 7,78% na contagem de leucócitos nas câmaras de Neubauer e de Nageotte, respectivamente. Três níveis do controle hematológico comercial foram utilizados na análise da exatidão, e todos os parâmetros obtiveram ER dentro do limite de ± 15% quando comparados aos valores de referência. Na comparação de métodos, as contagens foram realizadas em 60 amostras de sangue total. O viés calculado na análise de concordância de Bland-Altman foi de -0,0192 x 106/μL; 0,0386 x 103/μL e de 6,03 x 103/μL para as contagens de hemácias, leucócitos e plaquetas, respectivamente. Discussão Os resultados obtidos nas contagens realizadas no laboratório de controle de qualidade da FHB foram precisos e exatos apresentando coeficientes de variação e erros relativos baixos. O viés da análise de Bland-Altman para a contagem de hemácias e leucócitos foi próxima de zero e não estatisticamente significativa indicando que há concordância entre a contagem manual e a automática. Na contagem de plaquetas, o viés foi 6,03 x 103/μL e estatisticamente significativo indicando que na contagem manual foram contadas em média 6,033 x 103/μL plaquetas a mais que na contagem automática. Apesar de apresentar diferença estatística entre a contagem manual e a automática de plaquetas o erro relativo total entre ambas as contagens foi de 2,72%, estando em conformidade com o erro total aceitável para a contagem de plaquetas definido pela Clinical Laboratory Improvement Amendments (CLIA) em 25%. Conclusão As contagens manuais de hemácias, leucócitos e plaquetas realizadas no laboratório de controle de qualidade da FHB demonstraram precisão e exatidão, e obtiveram resultados comparáveis aos encontrados na contagem automática.

Introdução: O Linfoma não-Hodgkin (LNH) é uma forma comum de câncer hematológico que se origina de células B e T. Nos países ocidentais, o Linfoma Difuso de Grandes Células B (LDGCB) é o subtipo de LNH mais frequente, representando cerca de 31% de todos os linfomas em adultos. Desde a introdução da terapia antiretroviral (TARV), e subsequente reconstituição imunológica, o tratamento de pacientes HIV positivos é similar ao de pacientes que não convivem com o vírus, alcançando taxas semelhantes de sobrevida global (SG). Descrição do caso: Paciente, 38 anos, hipertenso, com obesidade grau III e portador de HIV em tratamento regular com TARV há 2 anos, apresentando massa em região rinossinusal com edema infraorbitário de crescimento rápido, sem sintomas B ou alterações neurológicas. Biópsia da lesão confirmou LDGCB, EBV positivo através de EBER, com acometimento do Sistema Nervoso Central (SNC) confirmado por imunofenotipagem do líquido cefalorraquidiano (LCR), sem infiltração da medula óssea. PET TC evidenciou múltiplas lesões líticas esparsas em todo o esqueleto axial e apendicular. Performance status ECOG 1, e International Prognostic Index (IPI) 3 pontos (risco intermediário/alto). Foi submetido a tratamento de 1ª linha com o protocolo R-CODOX-M/R-IVAC, com boa resposta clínica, e negativação do LCR. Cinco meses após o término do tratamento, ainda sem avaliação de PET-TC, apresentou aumento testicular bilateral, além de reaparecimento de edema infraorbitário à direita. USG e TC sugeriam envolvimento neoplásico de ambos os testículos. Realizou orquiectomia bilateral após congelação confirmar DGCB, consistente com refratariedade/recaída precoce ao tratamento. Iniciou protocolo de resgate com R-ESHAP com programação de quimioterapia intratecal e consolidação com Transplante Autólogo de Células Tronco Hematopoéticas mais radioterapia em bolsa escrotal. Discussão: O prognóstico do LDGCB melhorou substancialmente desde a incorporação do anticorpo monoclonal anti-CD20 aos protocolos convencionais de quimioterapia, atingindo taxas de SG em 5 anos de até 70%. Apesar dos avanços, uma minoria de pacientes com LDGCB sofrerá recidiva em sítios de santuário, SNC e/ou testículos, com resultados historicamente ruins. O risco é maior nos pacientes HIV positivos e/ou IPI elevado, e está associado a prognóstico ruim e sobrevida limitada a longo prazo. Conclusão: Pacientes com LDGCB e acometimento de santuário, refratários ou com progressão precoce, menos de 12 meses após o tratamento de primeira linha, constituem uma população de prognóstico reservado. Não há consenso sobre a melhor estratégia terapêutica de resgate. Idealmente esses pacientes deveriam ser encaminhados para pesquisa clínica, porém pacientes HIV positivos são tradicionalmente excluídos dos estudos, constituindo uma necessidade não atendida na Hematologia, especialmente no cenário público.

Background: Adult patients afflicted with Acute Lymphoblastic Leukemia (ALL) still represent a subset of poor prognosis, with a 5-year overall survival of around 46%. The development of new therapeutic approaches is of utmost importance to the oncologic routine and the search for targeted therapies is ever growing in the translational field. Objective: In this study, we aim to investigate an agonistic effect of poly-ADP-ribose polymerase 1 (PARP1) in inducing parthanatos, a PARP1-dependent and programed mode of cell death, in leukemic cell lines overexpressing PARP1, through inhibition of its antagonist, poly-ADP-ribose glicohydrolase (PARG). Methods and materials: We screened 6 cell lines for their PARP1 mRNA expression through qPCR. Inhibition of their metabolic activity was screened through Alamar Blue assay (Invitrogen™) after treatment with either a PARG inhibitor, JA2131, a PARP inhibitor, AZD2461, or a cytotoxic control in Doxorubicin. An IC50 that considered only cell death was assayed through treatment in serial dilutions followed by DAPI solution staining and analysis in flow cytometry. The profile of early death was determined by co-staining of treated cells with CellEvent™Caspase-3/7 Green Reagent (Invitrogen™), APC Annexin V (BD Pharmingen™) and DAPI Solution. Results and discussion: Of the initially screened cell lines, only B-ALL models significantly overexpresed PARP1 when compared to PBMC of healthy donors. Also, while all models responded to Doxorubicin treatment, only B-ALL models had measurable metabolic inhibition when incubated with PARGi and PARPi treatment. Cell death measured by DAPI has only been conducted in one cell line as of this moment, SUP-B15, a B-ALL model. While PARPi and Doxorubicin had IC50 values similar to those observed in metabolic inhibition by Alamar Blue, PARGi only induced cell death in treatment values approximately 3-fold higher than the necessary for metabolic inhibition. Nevertheless, cell death induced by the treatments demonstrate different early death profiles, with PARGi treatment not inducing any effector caspase activity previously to cell death, a hallmark of parthanatos, while PARPi and Doxorubicin had clear increase in caspase activity and annexin-V staining in early apoptotic cells. Conclusion: Targeted therapeutics with PARG or PARP inhibitors may be a feasible option for B-ALL treatment, with different modes of cell death induction. However, further analysis needs to be conducted in order to better characterize treatment-induced phenotypes and their cytotoxicity over non-neoplastic cells.

Acute Lymphoblastic Leukemia (ALL) is a hematological neoplasm characterized by clonal proliferation of lymphoblasts, being one of the most common types of leukemia in children and young adults, with a worse prognosis when manifested in adult patients. Despite advances in treatment, overall survival (OS) varies widely, influenced by several risk factors, such as cytogenetic characteristics, age, and disease subtype. This study seeks to analyze the impact of these factors on the overall survival of ALL patients, highlighting the importance of a personalized therapeutic approach to improve clinical outcomes. This retrospective study included 53 patients diagnosed with ALL treated at the Hospital Geral de Fortaleza (HGF), which is the largest oncohematology outpatient clinic in the state of Ceará. Patients were stratified according to the World Health Organization (WHO) guideline. The OS was assessed using Kaplan-Meier curves for each risk group and its’ time was defined as the period from diagnosis to death or the last follow-up. Statistical analyses were performed to compare the survival curves between the different risk groups, using the log-rank test to determine statistical significance. Of the 53 analyzed patients, 45 had B-ALL (median age of 40 years) and 8 had T-ALL (median age of 28 years). In all, 23 patients were considered to have high-risk cytogenetic and 30 were over 35 years old. The analysis showed that patients without high-risk cytogenetics (20% in 40 months) had a significantly lower survival compared to those with high-risk cytogenetics (50% in 40 months). This may seem counterintuitive, as high-risk features are expected to be associated with a worse prognosis. Perhaps this is due to the fact that patients with high-risk cytogenetics can receive more intensive or innovative treatments, leading to a better initial response and, consequently, longer survival. Age proved to be an important prognostic factor, with older patients having significantly lower survival (25% in 40 months). This is consistent with the literature, where it is known that advanced age is a risk factor for a worse prognosis in ALL, due to factors such as higher rates of comorbidities and resistance to treatments. Overall, we observed that patients with T-ALL showed slightly better survival compared to patients with B-ALL, particularly in the first 10 months. This may be explained because T-ALL may have a more favorable initial response to chemotherapy treatment compared to B-ALL. This study's results underline the importance of a personalized approach in the ALL treatment. It is evident that factors such as cytogenetics, age, and ALL subtype have a significant impact on patient overall survival. Stratification based on these factors can help in the selection of more appropriate treatments and in the definition of more accurate prognosis.

Acute myeloid leukemia (AML) is a hematologic malignancy characterized by the rapid proliferation of abnormal myeloid cells in the bone marrow and peripheral blood. Cytogenetics plays a crucial role in the risk stratification and prognosis of AML patients, according to the latest guidelines from the World Health Organization (WHO) and the European LeukemiaNet (ELN). This study aims to analyze the impact of high-risk cytogenetics on the survival of non-promyelocytic AML (non-APL) patients. In all, 93 non-APL AML patients were diagnosed between July 2021 and June 2024 at the Hospital Geral de Fortaleza (HGF), which is the largest oncohematology outpatient clinic in the state of Ceará. Survival data from non-APL AML patients were analyzed, stratified according to the presence or absence of high-risk cytogenetics, according to the guidelines. The analysis was performed using the Kaplan-Meier method, comparing two groups: patients with high-risk cytogenetics and patients without high-risk cytogenetics. Among the 93 patients, 47 were women and 46 were men, with a median age of 52.9 years (17-96 years). It was not possible to stratify all patients due to the lack of cytogenetic results. Among the available results, it was possible to stratify 15 patients with high-risk cytogenetic and 38 without high-risk cytogenetic. Herewith, a significant difference in the probability of survival was observed between the two groups over time, where the group with high-risk cytogenetics demonstrated a sharp drop in the probability of survival in the first 10 months, with a very low residual survival after this period, while the group that was not high-risk had a higher and more stable survival rate over time, with a probability of survival greater than 75% at 40 months. The results indicate that the presence of high-risk cytogenetics is associated with a significant worsening in overall survival of non-APL AML patients. Patients with high-risk cytogenetics show a rapid and marked decrease in the probability of survival in the first 10 months after diagnosis, underscoring the need for more aggressive and personalized therapeutic strategies for this subgroup. This occurs similarly in the literature that points to adverse cytogenetic abnormalities, such as in acute myeloid leukemia and chronic lymphocytic leukemias. These findings reinforce the importance of cytogenetic stratification in the clinical AML management. Patients with high-risk cytogenetics have a worse survival, which warrants more intensive therapeutic interventions and close monitoring. Cytogenetics-based risk stratification should be standard practice in treatment assessment and planning.

Introduction and objectives: The rs3824662 polymorphism in GATA3 is a risk variant for the development of B-cell precursor acute lymphoblastic leukemia (B-ALL), particularly BCR::ABL1-like ALL, a subtype of B-ALL defined by a gene expression profile similar to the observed in BCR::ABL1 positive patients. The predominance of high-risk alterations jointly with the absence of specific treatment protocols for adolescents and young adults (AYA) has negatively impacted the prognosis of this age subgroup. Besides that, studies evaluating the GATA3 SNP in AYA are rarely observed. Given this scenario, this study aims to evaluate the clinical and molecular impact of the SNP rs3824662 on the development of B-ALL in AYA. Material and methods: 76 samples from AYA patients (15 to 39 years old) diagnosed with B-ALL according to the World Health Organization criteria were included between 2018 and 2023. This project was approved by the INCA Research Ethics Committee (#87793418.0.0000.5274). The samples were obtained from different cancer treatment centers in Brazil. DNA was extracted using the DNA MiniBlood kit (Qiagen) following the standard protocol. The rs3824662 polymorphism was identified using the TaqMan allelic discrimination assay (C_27522049_10, Life Technologies). The observed genotypes were validated by direct sequencing using the ABI 3500 Genetic Analyzer (Life Technologies). The BCR::ABL1-like subtype was characterized by the presence of rearrangements in PDGFRB, CRLF2, JAK2, ABL1, ABL2, IGH, and/or CSF1R evaluated by FISH. GATA3 SNP was correlated with previously characterized clinical and molecular variables, including primary alterations detected by qPCR and additional alterations identified by the SALSA MLPA P335-C1 kit (MRC Holland). Results: We successfully analyzed 76 patients, displaying a median age of 19.5 years-old and the majority had a low white blood cell (WBC) count (77.0%). Regarding molecular subgroups, most of the patients did not present recurrent alterations and were characterized as B-other (35.9%). Heterozygote and mutant genotypes of rs3824662 SNP were observed in 39.7% and 8.2% of patients, respectively. The mutant allele A rarely occurred in concomitance with BCR::ABL1 or TCF3::PBX1, and was particularly overrepresented in B-other patients and BCR::ABL1-like subtypes (P < 0.05). Additionally, it was more common in patients with low WBC count (86.1%). Curiously, patients with the mutant genotype had a very low frequency of deletions in IKZF1, EBF1, JAK2, CDKN2A/B, PAX5, ETV6, BTG1, RB1, and deletions in the PAR1 region were not observed (P < 0.05). Discussion and conclusion: Our findings are consistent with previous studies in AYA B-ALL, corroborating that rs3824662 SNP is a significant factor for the development of BCR::ABL1-like B-ALL. Furthermore, there is an underrepresentation of this SNP in concomitance with deletions in genes involved in differentiation, proliferation, and cell cycle that are frequently affected in B-ALL. These findings suggest that this SNP can directly impact the development and progression of leukemogenesis, and could be incorporated in the diagnostic routine of B-ALL patients. Support: FAPERJ, CNPq, Ministério da Saúde, Swiss Bridge Foundation.

Introduction and objectives: B-cell precursor acute lymphoblastic leukemia (B-ALL) is characterized by a variety of genomic alterations relevant for risk stratification and prognosis of patients. Notably, Philadelphia-like (Ph-like) ALL is defined by a gene expression profile similar to Philadelphia-positive patients but without the historically known BCR::ABL1 rearrangement. The Ph-like subgroup is associated with an unfavorable prognosis and is observed in 15% of children and 40% of adolescents and young adults (AYA) with B-ALL. The characterization of this subgroup involves the evaluation of gene expression of 8 genes, and it is a complex challenge for centers of leukemia diagnosis. Given this scenario, the aim of this study is to establish a predictive genetic panel for defining the Ph-like subgroup in the Brazilian Unified Health System (SUS), thereby improving and expediting its diagnosis. Materials and methods: 108 samples from children and adult patients diagnosed with B-ALL according to World Health Organization criteria, and collected from different cancer treatment centers in Brazil were included. The patients were classified as B-other based on the absence of recurrent primary alterations: high-hyperdiploidy, ETV6::RUNX1, BCR::ABL1, TCF3::PBX1 and KMT2Ar. The Ph-like subgroup was characterized using cDNA samples to analyze the expression profile of SPATS2L, MUC4, CRLF2, CA6, NRXN3, BMPR1B, JCHAIN, and ADGFR1 genes by quantitative PCR (qPCR) using commercial TaqMan assays (Life Technologies). Gene expression profiles were correlated with the presence of rearrangements in genes frequently altered in this subgroup, including PDGFRB, CRLF2, JAK2, ABL1, ABL2, IGH and CSF1R, evaluated by Fluorescent in situ Hybridization (FISH) using break-apart probes (Zytovision and Cytocell). Patients positive for primary alterations were used as a reference control group. This study was approved by the Research Ethics Committee of INCA (CAEE #87793418.0.0000.5274). Results and conclusion: A total of 53 samples were included so far. Among them, 32 patients were classified as B-other and the remaining 21 patients had recurrent abnormalities: ETV6::RUNX1 (n = 6), BCR::ABL1 (n = 5), KMT2A-r (n = 3), TCF3::PBX1 (n = 3), and high-hyperdiploidy (n = 4). The Ph-like profile was identified in 37.5% of patients, based on the occurrence of CRLF2, ABL1 or JAK2 rearrangements and the expression profile, simultaneously. Most Ph-like patients were AYA (66.7%) and had low white blood cell count at diagnosis. It was observed that 11 of the 12 Ph-like patients had at least one overexpressed gene. Our preliminary results confirm the existence of distinct molecular profiles previously described among the subtypes of Ph+ and Ph-like B-ALL. While FISH has demonstrated effectiveness to identifying Ph-like cases when a differential gene expression was not observed, the current sample size is not enough to confirm the most appropriate diagnostic strategy for characterize this complex B-ALL subtype. Support: CNPq, FAPERJ, Ministério da Saúde, Swiss Bridge Foundation.

B-cell precursor acute lymphoblastic leukemia (B-ALL) is the most prevalent form of ALL and its development varies with age. It is characterized by chromosomal and genetic alterations such as aneuploidy, gene fusions, copy number alterations (CNA) and point mutations, which are crucial for diagnosis and therapy risk stratification. However, ̃25% of pediatric and ̃8% of adult cases with B-ALL lacking recurrent alterations are classified as “B-other”. New clinically relevant genetic alterations involving PAX5 gene are being demonstrated in this subtype, including rearrangements (PAX5 -r), deletions (PAX5 del), amplifications (PAX5 amp), and a point mutation (PAX5 P80R). Around 30% of B-ALL cases show PAX5 alterations (PAX5 alt), which are associated with relapses and poor outcomes. This study aims to characterize PAX5 alterations and evaluate their prognostic impact in children, adolescents and young adults (AYA) with B-other ALL. Pediatric (< 15 years) and AYA (15-39 years) cases diagnosed with B-ALL between 2018 and 2023 in different Brazilian cancer treatment centers were included. Recurrent and primary alterations were previously characterized by reverse transcription PCR (RT-PCR) or quantitative PCR (qPCR). Only patients categorized as B-other, defined by the lack of ETV6::RUNX1, BCR::ABL1, TCF3::PBX1, KMT2A::AFF1 and high-hyperdiploidy (DNA index), were included. CNA were identified by multiplex ligation-dependent probe amplification (MLPA), using the P335-C1 probemix (MRC-Holland). PAX5 -r were detected by fluorescence in situ hybridization (FISH - ZytoLight SPEC PAX5 Dual Color Break Apart Probe - Zyto Vision), and the PAX5 P80R mutation was assessed by TaqMan customized allelic discrimination assay (Life Technologies). This study was approved by the institutional ethical committee (CAEE #87793418.0.0000.5274). A total of 62 B-other ALL cases were evaluated (44 pediatric and 18 AYAs). Approximately 27% of these cases were characterized as PAX5 alt, due to the presence of PAX5 amp (̃23%), PAX5 del (̃52%) or PAX5 -r (̃41%), occurring alone or simultaneously. In this subtype, most cases were pediatric (71%), with a median age of 8 years-old, and low white blood cell count at diagnosis (̃64%). CDKN2A/B del (̃47%) and JAK2 del (̃17%) were concomitantly observed with PAX5alt. On the other hand, in PAX5 wild-type patients, CDKN2A/Bdel (̃26%) and IKZF1 del (20%) were the most prevalent. Curiously, the IGH -rearrangements were commonly found among PAX5 alt cases (̃23%). PAX5 P80R were not found in any cases. Considering that the genetic alterations knowledge is essential for the treatment decisions, a better understanding of PAX5 alterations in B-other ALL is crucial for precision medicine. Besides, our findings suggest that the IGH gene may be a frequent partner of PAX5 fusions, though it requires further investigation.

Introdução: A neoplasia de células dendríticas blásticas plasmocitóides (NCDBP) é rara, seus principais locais de envolvimento são a pele e medula óssea, com frequente envolvimento do sistema nervoso central (SNC). O acometimento ocular é raramente descrito. O prognóstico é reservado e o tratamento baseia-se em protocolos de quimioterapia (QT) utilizados no tratamento de leucemias agudas ou linfomas, seguido de transplante alogênico de medula óssea (aloTMO) em pacientes elegíveis. Objetivo: Descrever as características clínicas, protocolos de tratamento e evolução dos pacientes portadores de NCDBP com acometimento ocular tratados no INCA. Material e métodos: Foram identificados 2 pacientes. Os dados foram colhidos do prontuário físico e eletrônico. O diagnóstico foi através de biópsia cutânea. Resultados: Caso 1: Homem, 62 anos, iniciou quadro de lesões cutâneas nódulo-tumorais progressivas em 80% da superfície corporal, linfonodomegalias e citopenias, com evolução de 6 meses. Apresentava massa subconjuntival próxima à carúncula do olho esquerdo medindo 1 cm em sua maior extensão, clinicamente semelhante as lesões cutâneas, a biópsia de pele mostrou denso infiltrado dérmico composto por células blásticas, positivas na imunohistoquímica para CD123, CD4 e CD56. A medula óssea tinha infiltração neoplásica com marcadores semelhantes as lesões cutâneas e sugestivos de NCDBP. O paciente iniciou protocolo Hiper-CVAD (ciclofosfamida, vincristina, doxorrubicina e dexametasona) alternando com altas doses de metotrexato e citarabina, obteve resposta parcial, com redução significativa da lesão ocular, entretanto, faleceu 73 dias após o diagnóstico por recaída doença e refratariedade à QT. Caso 2: Mulher, 52 anos, apresentou múltiplas lesões nodulares violáceas evolutivas. A biópsia cutânea mostrou infiltração dérmica difusa com células blásticas, imunofenótipo CD4+ CD123+ CD56+. Não havia envolvimento da medula óssea em diagnóstico. O tratamento eleito foram seis ciclos de hiper-CVAD alternados com altas doses de metotrexato e citarabina que resultou em remissão completa. Durante a manutenção oral com mercaptopurina e metotrexato, ela apresentou recidiva isolada no braço direito, e posteriormente desenvolveu múltiplas lesões tumorais, paralisia facial periférica direita e uma lesão subconjuntival violácea elevada do saco inferior. A punção lombar revelou células blásticas com morfologia monocitóide, microvacúolos e projeções citoplasmáticas. Foi feito o diagnóstico de recidiva sistêmica, ocular e do sistema nervoso central de BPCDCN. A paciente decidiu não prosseguir com tratamento e evoluiu para o óbito 8 meses após o diagnóstico. Discussão: Os dois pacientes apresentavam lesões cutâneas ao diagnóstico e tiveram acometimento neoplásico ocular. Sendo o primeiro caso desde o diagnóstico, feito mais tardiamente, e o segundo em sua recidiva. Tratados com protocolos de QT semelhantes, evoluiram com desfecho desfavorável e óbito precoce. O tratamento de escolha para indução de remissão da NCDBP é o Tagraxofusp, porém essa medicação não se encontra disponível no Brasil, onde os pacientes recebem preferencialmente protocolos baseados em QT, seguido de aloTMO, quando elegíveis. Entretanto, esse tratamento tipicamente tem curta duração de remissão. Conclusão: A NCDBP com acometimento ocular, é uma doença rara e agressiva, pouco descrita na literatura. O seu prognóstico é reservado, o tratamento tem resultados desapontadores, sendo uma necessidade médica não atendida.