bluuebunny/crossref_metadata_2025_split huggingface dataset:
EP102: Pharmacological Inhibition of METTL3 Elicits Tumor Growth Inhibition In Vivo and Demonstrates Synergy with Venetoclax in Various AML Models
Authors: Fraser Graeme , Sorlet Catherine , Parmentier Nicolas , Hartiel Anne-France , Hospied Sandrine , Mompied Clement , Absil Genevieve , Oukoloff Killian , Dutheuil Guillaume 
Published: December 2023

METTL3 is the RNA methyltransferase predominantly responsible for the addition of N-6-methyladenosine (m6A), the most abundant modification to mRNA. The prevalence of m6A and the activity and expression of METTL3 have been linked to the appearance and progression of acute myeloid leukemia (AML) 1. EPICS has discovered and optimized a small molecule inhibitor of METTL3 (“M3i”). M3i was shown to inhibit the enzymatic activity of METTL3 (IC 50 = 1.8 nM, SPA assay). The anti-proliferative effects of M3i was demonstrated in various AML cell lines (Kasumi, IC 50 = 63 nM; MV-411, IC 50 = 506 nM; KG1a, IC 50 = 99 nM). Biomarkers were evaluated following M3i treatment of Kasumi and MV-411 cells with consistent results demonstrating depletion of oncoproteins relevant to apoptotic (BCL-2, MCL1) and proliferative (MYC, BRD4, SP1) pathways. In addition, combined treatment with M3i and venetoclax showed clear synergistic effects 2 to inhibit proliferation on both cell lines consistent with the depletion of BCL-2 and MCL1, known targets for venetoclax activity and resistance mechanisms. M3i has been optimized for oral dosing with oral exposure confirmed in mouse, rat and minipig (oral bioavailability &gt;80%, rat; &gt;30%, minipig). In vivo efficacy was evaluated in a disseminated xenograft model using established systemic MV-411-Luc-mCh-Puro 3 in female NSG mice where M3i (10, 30 mg/kg, oral, QDx31 days) demonstrated dose-dependent tumor growth inhibition (p&lt;0.01, relative to vehicle control) of cancer progression as measured by in-life imaging and confirmed by flow cytometry demonstrating significant, dose-dependent pharmacological knockdown of hCD45+ cells in bone marrow, blood and spleen (p&lt;0.01, in all cases). These results confirm that the pharmacological inhibition of METTL3 arrests proliferation of AML and further provide a mechanistic basis for the synergistic activity of M3i in combination with venetoclax as is a viable strategy for the treatment of disease.
1Yankova E et al. (2021), Nature 593:597-601; 2Ianevski A et al. (2022), Nucl Acids Res 50:W739-W743; 3Trachet E (2016), LabCorp Drug Development website: model spotlight MV (4;11)
Comparison of Live Cell Enrichment Methods for Single-Cell Rnaseq
Authors: Meskauskaite Urben Brigita , Krause Tobias , Takizawa Meril , Snopkowski Catherine , Masilionis Ignas , Remsik Jan , Parisotto Yollanda , Brown Chrysothemis , Boire Adrienne , Pe'er Dana , Sharma Roshan , Chaligne Ronan 
Published: December 2023

Single-cell RNA sequencing (scRNA-seq) has emerged as a powerful tool for investigating cellular diversity and gene expression profiles at the individual cell level. However, to ensure accurate analysis of scRNA-seq data, it is crucial to generate high-quality single-cell suspensions by effectively removing dead cells, debris, and ambient RNA. This study aims to assess and compare the performance of five commercially available methods for dead cell removal: Fluorescence activated cell sorting (FACS), Akadeum Microbubble kit, StemCell EasySep kit, Miltenyi Biotec Dead Cell Removal Kit, and LevitasBio Levicell Live Cell Enrichment.
Among the 5 methods we evaluated, FACS is considered the gold standard technique due to its remarkable efficiency and versatility. However, the high-pressure nature of FACS can exert mechanical stress on cells during the sorting process, potentially affecting cellular viability and gene expression profiles. On the other hand, the remaining four methods, while less versatile, utilize milder conditions, which may better preserve fragile cell populations.
To evaluate those 5 methods, we used two tissues requiring only no to mild dissociation process: lymph node and cerebrospinal fluid (CSF). Viability of cells from each tissue was assessed using trypan blue staining after dissociation. The cell suspensions were then subjected to the five dead cell removal methods following the manufacturer's instructions. For each cell suspension, 10,000 cells were targeted using the 10x Genomics 3' v3.1 kit. The evaluation criteria encompassed hands-on time, required equipment, cost, improvement in live cell percentage, cell type recovery, gene expression profile, and cellular stress response.
We hope these findings will provide valuable guidance for researchers in selecting the most suitable dead cell removal method based on their experimental requirements, thereby facilitating efficient, cost-effective, and high-quality scRNA-seq analysis.
Quantitative Ki-67 Proliferative Index Assessment of Human B-Cell Lymphomas By Flow Cytometry
Authors: Castillo Felipe , Morales Constanza , Iruretagoyena Mirentxu , Díaz Schmidt Joaquin , Ernst Daniel 
Published: December 2023

Background:
The Ki-67 proliferative index is used as a diagnostic and prognostic marker in the classification of B-cell non-Hodgkin lymphomas (NHL). Despite its semiquantitative nature, immunohistochemistry (IHC) evaluation of Ki-67 in histologic specimens is the gold standard. The aim of this study was to determine the correlation of quantitative Ki-67 expression by flow cytometry (FCM) with standard IHC in samples of B-cell NHL.
Methods:
We performed a retrospective observational study of bone marrow and tissue biopsies evaluated for routine lymphoma screening between January 2021 and December 2022. FCM analysis was performed according to Euroflow standard LST screening. Positive samples were further characterized, and Ki-67 expression was integrated using a PerCP-Cy5.5 fluorochrome-conjugated monoclonal antibody after cell fixation and permeabilization. The Ki-67 proliferative index was calculated as all positive events among the neoplastic population (%). Ki-67 proliferative index by flow cytometry was compared with Ki-67 proliferative index by IHC. Clinical data were retrieved from the electronic database. This study was approved by the institutional IRB and clinical ethics committee.
Results:
A total of 51 samples from 50 patients were evaluated: 4 bone marrow aspirates, 8 fine-needle aspirations (FNA) and 39 tissue biopsies. Median age at diagnosis was 67 years (interquartile range [IRQ], 60-79 years), and 45% were female. A low-grade lymphoma was diagnosed in 27 cases (52.9%; 20/27 follicular lymphoma), a high-grade lymphoma case in 23 cases (45.1%; 20/23 diffuse large B-cell lymphoma), and mantle cell lymphoma in 1 case.
Median Ki-67 proliferative index by FCM was 41.4% (range, 1 - 100%) and 54.8% (range, 5 - 100%) by IHC. Overall, the Ki-67 proliferation index by FCM showed a significant correlation coefficient of 0.925 with IHC (95% IC 0.870 - 0.957, p &lt; 0.0001). There was a negative constant bias of -13.35% (-46.87 to 20.16), which was greater in the range of 30-80% of Ki-67 expression by IHC.
Indolent lymphomas displayed a lower proliferative index by FCM compared with IHC (5 vs 20%, p&lt;0.001). No difference was seen among aggressive lymphomas. Fourteen cases had a difference in the proliferative index &gt;20%, 10/14 were follicular lymphoma, 2/14 were indolent lymphoma transformations and 2 were large B-cell NHL. In all cases FCM displayed lower results compared with IHC.
Conclusions:
Our data suggest that the Ki-67 proliferative index can be accurately assessed by flow cytometry. A correlation bias seems clinically meaningful in intermediate values of Ki-67. The quantitative nature of FCM-based proliferative index may improve the prognostic potential of Ki-67, especially for indolent or borderline histologies. Further validation studies are warranted.
The Biodistribution of the Novel Heme and Hemoglobin Scavenger Apohemoglobin-Haptoglobin Complex in Healthy C57BL/6 Mice
Authors: Lucas Daniela , Munoz Carlos J , Muller Cynthia R , Gopal Srila , Cabrales Pedro 
Published: December 2023

Background: Apohemoglobin-haptoglobin (apoHb-Hp) complex represents a bioengineered complex designed to scavenge hemoglobin and heme. These substances are released into the intravascular space during hemolysis, leading to vasoconstriction, inflammation, and disease progression. The use of apoHb-Hp complex has already demonstrated promising results in sickle cell and beta-thalassemia mouse models, highlighting its potential as a therapeutic agent. However, there are still crucial aspects that remain poorly understood, particularly concerning the biodistribution and pharmacokinetics associated during chronic administration. Understanding how the drug is distributed throughout the body is essential for optimizing its therapeutic potential, ensuring patient safety, and guiding both drug development and clinical use.
Methods: To address these critical questions of apoHb-Hp biodistribution and pharmacokinetics, we conducted a study using twenty C57BL/6 mice from Jackson Laboratory. The mice were treated with the apoHb-Hp complex via daily injection every other day for different periods: 24 hours, one week, three weeks, and six weeks, with five mice in each subgroup. The dosage used was 80 mg/kg, and the concentration was 33.6 mg/mL. To evaluate the half-life and biodistribution of the complex, the last dose in each subgroup was radiolabeled with technetium-99m (99mTc-ApoHb-Hp).The intravascular 99mTc-ApoHb-Hp levels were measured after 30 minutes, 1 hour, 4 hours, and 24 hours to establish the drug's half-life. After the final tail vein sample collection, the mice were sacrificed, and their hearts, livers, spleens, lungs, and kidneys were weighed and collected to determine the amount of 99mTc-ApoHb-Hp present in each organ.
Results: The results indicate that a significant portion of the 99mTc-ApoHb-Hp was accumulated in the liver, and regardless of the duration of dosing, the half-life for the 24 hour group was 10 hours, for the 1 week it was 6.5hrs, and for the 3 weeks it was 9hrs, showing no significant difference between the groups. hours. Liver accumulation favors the hepatocytes metabolism degradation of the apoHb-Hp complex from the body. There was no significant differences in half life based on dosing chronicity, likely suggesting that there was no additional degradation of ApoHb-Hp by immune mediated mechanisms such as antibody formation.
In conclusion, Biodistribution of ApoHb-Hp was predominantly in the liver in both acute setting ( single dose) and chronic setting ( multiple doses). The average half life of the infusion was 5 hours and the predominant clearance mechanism was likely via hepatic elimination. Further research and development may help improve our understanding of the biodistribution of the apo Hb- Hp complex, therapy opening new doors for the treatment of hemolytic anemias using the ApoHb-Hp complex.
Radiation Therapy in the Management of Primary Gastric Diffuse Large B-Cell Lymphomas: Long-Term Outcomes of 82 Patients
Authors: Fregonese Beatrice , Hajj Carla , Gonzalez Silvano , Lee Jisun , Boe Lily , Imber Brandon S. , Yahalom Joachim 
Published: December 2023

Background/Objectives: Our group has recently published its long-term experience treating gastric MALT lymphomas with involved site radiation therapy (ISRT) and supported the use of RT alone as standard of care for H pylori independent MALT. For primary gastric (PG) diffuse large B-cell lymphomas (DLBCL), R-CHOP is the mainstay of therapy, but RT to the stomach primarily as consolidation has rarely been analyzed. In this study, we focus on pts with PG DLBCL who received combined modality therapy.
Materials/Methods: 285 consecutive pts with PG lymphomas treated with RT between 1/2000-1/2021 at our institution were retrospectively identified. Baseline characteristics and follow-up data were abstracted. Overall response rate (ORR) was assessed with Lugano PET criteria and endoscopy. Overall survival (OS) was calculated using Kaplan-Meier. Toxicity was assessed with CTCAE v4.0.
Results: Of 285 pts, 203 (71%) had PG MALT and 82 (29%) PG DLBCL. Baseline characteristics are summarized in Table 1. In the DLBCL pts, 74 (90%) received consolidation RT after limited (3-4 cycles) or full (6 cycles) chemoimmunotherapy (CT), 4 (5%) received RT after 2 lines of CT, 2 (3%) received first line RT followed by CT, 1 (1%) had 3 lines of CT and RT, and 1 (1%) was treated with surgery, 2 lines of CT, and RT. In the 74 pts who received CT+RT as first line treatment, the median RT dose was 3060 cGy (range 1620-3600). Based on post-chemo pre-RT EGD and PET, 61 (82%) pts were in CR, 7 (10%) had PR, 3 (4%) had SD and 3 (4%) pts were not assessed. Overall, 66 (89%) pts who received RT after CT achieved a CR. Of the remaining 8 (11%) pts who did not achieve a CR after RT, 4 had PR, 1 had SD, 1 was lost to FU, 1 started additional CT before response assessment and 1 pt died. The 2 pts in the DLBCL group who received first line RT achieved CR after RT. Side effects after RT were mild and limited to nausea and fatigue. The median follow-up time was 5.4 years (95% CI 0.4, 20.4) in DLBCL pts and 6.3 yrs (95% CI 0.3, 22.1) in MALT pts. The 5-year OS was 80% (95% CI 71-90%) for DLBCL and 96% (95% CI 93-99%) for MALT.
Conclusion: Adding stomach RT following chemoimmunotherapy is feasible, well-tolerated and safe. More in depth analysis PG DLBCL is underway, with a plan to compare outcomes of patients treated with CT alone versus CT followed by RT, and further analyze treatment-related toxicities, progression-free survival, and patterns of recurrence.
Changes in Immune-Related Indicators in Patients with Hematopoietic Abnormalities and Their Impact on Clinical Symptoms
Authors: Chen Mengran , Zhao Lei , Yang Jinjun , Wu Yu 
Published: December 2023

Background: The pathogenesis of myelodysplastic syndrome (MDS) is mainly driven by specific genetic lesions involving stem cell compartments. But recent studies have found that MDS also appears to be related to immune mechanisms. There is still no consensus on which immune pathway is involved. As a result, we investigated the potential implications of various immune cell subpopulations on hematopoietic abnormalities disease occurrence and development.
Methods: 69 MDS patients in our hospital and 31 normal controls are involved. Clinical data were collected and immune-related indicators were detected through blood routine testing, flow cytometry, ELISA, and other testing methods. Then we analyzed the differences between immune groups in patients and normal individuals, as well as the correlation between the immune group and clinical symptoms.
Results: In lymphocytes, the activated T cell population was significantly increased in patients with MDS (p&lt;0.01), with CD4 + T cell being the most significantly elevated (p&lt;0.01). In atypical T cell subtypes, NK-T was the most significantly reduced (p&lt;0.01). Among immune cell receptors, the NK-P46 was significantly increased (p&lt;0.01). And there was a significant decrease in immune cell activity experiments (p&lt;0.01). In the correlation study, we found a negative correlation between neutrophils and NK-NKG2D cells (r=-0.52, p&lt;0.01), and a positive correlation between bone marrow progenitor cells and Th17 (r=0.47, p&lt;0.05). (figure 1)
Conclusion: It has been confirmed that the immune mechanism has indeed had an impact on MDS. We found a significant increase in the T cell population but a decrease in immune cell function in patients with MDS. Indicating a new direction for studying immune mechanisms in MDS patients.
Management and follow-up of pregnancy-onset thrombotic thrombocytopenic purpura: the French experience
Authors: Béranger Nicolas , Coppo Paul , Tsatsaris Vassilis , Boisseau Pierre , Provôt François , Delmas Yahsou , Poullin Pascale , Vanhoorelbeke Karen , Veyradier Agnès , Joly Bérangère S. 
Published: December 2023

Abstract
Pregnancy-onset thrombotic thrombocytopenic purpura (TTP) is a rare and life-threatening disease of which diagnosis and management requires experienced multidisciplinary teams. The mechanisms responsible for a deficiency in the disintegrin and metalloprotease with thrombospondin type 1 repeats, member 13 (ADAMTS13) leading to pregnancy-onset TTP may be congenital or acquired, and studying ADAMTS13 conformation could be of interest. The differential diagnosis between TTP and other pregnancy-associated thrombotic microangiopathies (TMA) is often challenging. Our retrospective multicenter study highlights the significance and the challenges associated with pregnancy-onset TTP and childbirth in terms of diagnosis, obstetric management, and follow-up aspects. Among 1174 pregnancy-onset TMA enrolled in the French Registry for TMA from 2000 to 2020, we identified 108 pregnancy-onset TTP: 52 immune-mediated TTP (iTTP, 48.1%), 27 acquired TTP of unidentified mechanism (uTTP, 25%), and 29 congenital TTP (cTTP, 26.9%). Data show that maternal outcome is good (survival rate: 95%) and fetal outcome is linked to the gestational age at the onset of the disease (survival rate: 75.5%). Three distinct entities with different natural histories emerged: pregnancy-onset iTTP appears similar to idiopathic iTTP, with an open ADAMTS13 conformation, and is marked by a relapse risk independent of subsequent pregnancies; pregnancy-onset uTTP appears to have a different pathophysiology with an unexpected open ADAMTS13 conformation and a very low relapse risk independent of subsequent pregnancies; finally, pregnancy-onset cTTP is characterized by the necessity of pregnancy as a systematic and specific trigger and a need for prophylactic plasmatherapy for subsequent pregnancies. This trial was registered at www.clinicaltrials.gov as #NCT00426686, and at the Health Authority and the French Ministry of Health (P051064/PHRC AOM05012).
Psychological characteristics and emotional difficulties underlying school refusal in adolescents using functional near-infrared spectroscopy
Authors: Li Gaizhi , Niu Ying , Liang Xiumei , Andari Elissar , Liu Zhifen , Zhang Ke-Rang 
Published: December 2023

Abstract
Background
This study aims to explore the psychological characteristics, related emotional problems and potential NIR brain function mechanism of adolescents who refuse to attend school.

Methods
The study included 38 adolescents (12–18 years old) who were not attending school and 35 healthy controls (12–18 years old) who are attending school regularly. Participants completed (1) general demographics, (2) Eysenck Personality Questionnaire (EPQ), (3) Zung Self-Rating Depression Scale (SDS), (4) Zung Self-Rating Anxiety Scale (SAS), and (5) Symptom Checklist-90 (SCL-90). In addition to the clinical tests, participants completed functional near-infrared spectroscopy (fNIRS). Mental health, personality, and emotional state were evaluated in both groups to explore the differences and to understand the underlying mechanisms of school refusal during adolescence.

Results
Adolescents who did not attend school had higher neuroticism scores on the Eysenck Personality Questionnaire than healthy controls (p(FDR) < 0.001), introversion and concealment scores were lower than those of healthy controls (p(FDR) < 0.001), there was no significant difference in psychoticism scores between groups. SDS, SAS, SCL-90 scores and factor scores were higher than those of healthy control group (p(FDR) < 0.001), NIR functional brain imaging was different from healthy control group in the 12 and 27 channels (p(FDR) = 0.030, p(FDR) = 0.018), and no difference was found in the remaining channels (p(FDR) > 0.05). There were statistically significant differences in age and gender between the adolescents who refused school and the control group (p(FDR) < 0.001).

Conclusion
School refusal adolescents are relatively introverted and sensitive and need more attention in daily life. Although the adolescents’ emotional problems did not reach the diagnostic criteria of depressive disorder and anxiety disorder, their scores were still higher than those of the control group, suggesting that we should pay more attention to their emotional problems in order to better help them return to school. Using fNIRS, it was found that abnormalities in frontal lobe regions in adolescents with school refusal behaviors, which would contribute to early diagnosis and timely intervention of school refusal behaviors.
Prevalence of and factors associated with scrub typhus exposure among the hill tribe population living in high incidence areas in Thailand: a cross-sectional study
Authors: Tasak Nidanuch , Apidechkul Tawatchai , Law Andrew C. K. , Abdad Mohammad Yazid , Srichan Peeradone , Perrone Carlo , Tanganuchitcharnchai Ampai , Wongsantichon Jantana , Blacksell Stuart D. 
Published: December 2023

AbstractBackgroundScrub typhus is a significant tropical disease, occurring in rural settings and therefore usually afflicting remote agricultural populations who have lower socioeconomic status and limited access to medical care. A large proportion of the hill tribe people in Thailand are financially poor, have limited education, and do not have adequate health care access. This study aimed to estimate the prevalence of and determine factors associated with scrub typhus exposure among the hill tribe population living in high-incidence areas in northern Thailand.MethodsA cross-sectional study design was used to gather information from hill tribe people aged 18 years and over living in ten hill tribe villages in Mae Fah Luang, Chiang Rai Province, Thailand. Participants who met the inclusion criteria were invited to participate in the study. A validated questionnaire was used as the research instrument, and 5 mL blood samples were taken.Orientia tsutsugamushiIgM and IgG antibodies were detected by enzyme-linked immunosorbent assay (ELISA) and then confirmed by immunofluorescence assay (IFA). Logistic regression was used to detect associations between variables at a significance level of α = 0.05.ResultsA total of 485 hill tribe people participated in the study; 57.1% were female, 29.9% were over 60 years of age, 46.4% were from the Akha tribe, and 74.2% had never attended school. The overall prevalence of scrub typhus exposure was 48.0%. In the multivariate model, five variables were found to be associated with scrub typhus exposure. Participants aged over 60 years had a 4.31-fold increased risk (95% CI = 1.73–10.72) of scrub typhus exposure compared to those who were younger than 30 years. Those who were illiterate had a 3.46-fold increased risk (95% CI = 1.93–6.21) of scrub typhus exposure than those who had at least a primary education level. Participants from the Akha tribe had a 2.20-fold increased risk (95% CI = 1.31–3.72) of scrub typhus exposure than those from the Lahu tribe. Subjects who had a history of cutting grass had a 1.85-fold increased risk (95% CI = 1.20–2.84) of scrub typhus exposure. Those who never wore gloves for farming had a 2.12-fold increased risk (95% CI = 1.28–3.49) of scrub typhus exposure than those who wore gloves daily.ConclusionsThere is a high prevalence of scrub typhus exposure among the hill tribe in Thailand. Effective public health interventions to promote scrub typhus awareness and prevention are urgently needed in these populations.