2nd Generation Research Articles

Abstract 4061: Preclinical characterization of MBS309, a conditionally active PD-1-dependent IL-2 mutation with significantly superior anti-tumor efficacy with safety

Abstract Although PD-1/PD-L1 therapies have shown significant benefit among a range of tumor types, the majority of malignancy patients do not respond or develop resistance after initial tumor regression. Development of new modality to overcome primary and acquired resistance to anti-PD-1/PD-L1 therapy is a huge unmet medical need, and IL-2 is believed a potentially powerful approach for that. However, the current clinical application of IL-2 in cancer immunotherapy has been limited by its severe toxicity. Herein, we developed MBS309 (PD1/IL2m), a novel PD-1-targeted IL-2 mutation with significantly enhanced therapeutic window. MBS309 has the similar high affinity and blockade efficacy with monoclonal antibody to PD-1, and its IL-2 mutation has reduced binding to IL-2Rβγ and abolished binding to IL-2Rα. Our in vitro assays, including HEK-blue assay and CD3+ T cell assay, both indicated that the IL-2 receptor agonist activity of MBS309 was conditionally induced only the antibody cis-binding to both PD-1 and IL-2 Rβγ. And the maximal agonist activities of MBS309 in HEK-blue assay and CD3+ T cell assay were much lower than wild-type IL-2 and RG6279 analog enabling its high tolerated drug doses in vivo. In our in vivo tumor murine models, 1mg/kg MBS309 showed much better suppression effect in MC38/hPD-L1 tumor than 1mg/kg RG6279 analog and Pembrolizumab (1mg/kg) combination with wild type IL-2 (0.1mg/kg) ; And MBS309 at the dosage of 5mg/kg can significantly inhibit tumor growth in PD-1 resistant Pan-02 xenograft models without any side-effects observed, while RG6279 analog at the dosage of 5mg/kg induced severe body reduction of the mice and resulted in 80% (4/5) mice death after two dosages. MBS309 showed similar PK profile with anti-PD-1 monoclonal antibody and much better than RG6279 analog in mouse model. 20mg/kg of MBS309 was well tolerated in mouse, while 10mg/kg of RG6279 analog or PF-07209960 analog show severe toxicity and resulted in death of all the mice. Pharmacodynamic studies in murine models showed that MBS309 prioritized active tumor infiltrated PD-1+ CD8 T cells rather than other organs. Taken together, MBS309 effectively expands tumor infiltrated T-cells, and avoids system immune cytokine side effects compared to other PD-1-targeted 2nd generation IL-2Rβγ agonist modalities currently in clinical development. And it shows much higher anti-tumor effect compared to anti-PD-1/-L1 antibodies alone in both PD-1 sensitive and resistant tumor models. MBS309 has demonstrated a favorable safety profile and highly anti-tumor activities in vivo, which has the potential to supporting its clinical development for the treatment of cancer. Citation Format: Jiangmei Li, Guangzhong Lin, Lunfeng Zhang, Feng Li. Preclinical characterization of MBS309, a conditionally active PD-1-dependent IL-2 mutation with significantly superior anti-tumor efficacy with safety [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4061.

Abstract 44: A novel, codon-based method to modulate off-tumor toxicity of chimeric antigen receptor (CAR) T cells

Abstract Chimeric antigen receptor (CAR) T cell therapies for hematologic malignancies, while promising and efficacious, can also result in a variety of adverse effects. Of note, on-target, off-tumor toxicity is one adverse effect that continues to limit the safety and efficacy of CAR-T cells and has even led to life-threatening or fatal outcomes in patients (Morgan et al. 2010.). To address this, we have developed a unique, codon-based approach by replacing strategically selected codons within CAR non-antigen binding domains with their rare synonymous isoforms. Hoekema et al. (1987) reported the ability of synonymous rare codons to alter gene and protein expression in other model systems. In CAR-T cells, we aim to use this novel, codon-based approach to fine tune and modulate CAR-T cell expression and function to elicit full on-tumor activity while lessening off-tumor toxicity. In our approach, we replaced specific endogenous codons within the non-antigen binding domains of Hu8F4-CAR, a TCR-like 2nd generation CAR developed by our group which targets the leukemia-associated antigen PR1/HLA-A2 (Ma et al. 2016.). Briefly, we created variants of Hu8F4-CAR containing rare codon substitutions in two subtypes: intracellular-based (type 1) and combined extracellular- and intracellular-based (type 2). Each variant subtype (type 1 or 2) had either between 6 and 30 or 27 and 57 rare codons introduced, respectively. Expression profiling and functional assessment of the Hu8F4-CAR variants was assessed in the Jurkat mutant cell line (J.RT3-T3.5) or human primary healthy peripheral blood mononuclear cells (PBMCs) that were enriched for T cells, respectively. Expression profiling results show that surface expression is reduced in J.RT3-T3.5 cells when assessed by flow cytometry by 1-1.5-fold compared to unmodified Hu8F4-CAR. When evaluated by immunoblotting, novel lower molecular weight bands emerge only in rare codon-modified variants, suggesting that rare codons may have altered translational or post-translational processing leading to truncated CAR protein isoforms (30, 45, or 60kDa vs 73kDa for unmodified Hu8F4-CAR), which may contribute to reduced cell surface CAR expression. Furthermore, we have conducted in vitro functional testing of the rare codon-modified variants and show reduced activity against non-malignant off-tumor targets that express PR1 and/or HLA-A2 including healthy donor PBMCs, mobilized hematopoietic cells, and bone marrow. Importantly, in addition to reduced off-tumor activity against healthy myeloid cells, rare codon-modified variants preserve their activity against on-tumor target cells, including the U937-A2+ cell line and primary acute myeloid leukemia. Altogether, these results suggest that our approach can modulate CAR expression and functionality creating a novel methodology to reduce the likelihood of off-tumor adverse effects typically associated with CAR therapies. Citation Format: Rolando A. Vedia, Sijie Lu, Anne V. Philips, Anna Sergeeva, Lisa S. St John, Karen Clise-Dwyer, Gheath Alatrash, Jeffrey J. Molldrem. A novel, codon-based method to modulate off-tumor toxicity of chimeric antigen receptor (CAR) T cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 44.

Abstract 4651: A macrocyclic kinase inhibitor overcomes the compound mutations that cause the resistance to all approved EGFR inhibitors in EGFR-positive lung cancer

Abstract EGFR mutant cancers account for 20-30 % of non-small cell lung cancers and show marked sensitivity to EGFR tyrosine kinase inhibitors (TKIs). However, the tumor almost inevitably relapses due to acquired resistance. To date, several EGFR-TKIs have been approved, and the third-generation EGFR-TKI, osimertinib, has been widely used both as first-line therapy and for patients with the EGFR-T790M mutation after 1st or 2nd generation EGFR-TKI therapy. However, the EGFR-T790M+C797S compound mutation confers resistance to all approved EGFR-TKIs. In our previous study, we experimentally demonstrated that brigatinib with anti-EGFR antibody was effective against osimertinib-resistant EGFR-C797S and EGFR-T790M+C797S mutants, and is currently undergoing clinical studies. However, the tumor would relapse because of the acquisition of additional resistance mutations. Here, we first demonstrated the binding mode of brigatinib to the EGFR-T790M/C797S mutant by crystal structure analysis and predicted brigatinib-resistant mutations through a cell-based assay, including N-ethyl-N-nitrosourea (ENU) mutagenesis. We found that clinically reported L718 and G796 compound mutations appeared, consistent with their proximity to the binding site of brigatinib, and brigatinib-resistant quadruple mutants, such as EGFR-activating mutation/T790M/C797S/L718M, were resistant to all clinically available EGFR-TKIs. BI-4020, a fourth-generation macrocyclic EGFR inhibitor plus anti-EGFR antibody, overcomes the quadruple and major EGFR-activating mutants in vitro and in vivo, but not the minor mutants such as L747P. Microsecond molecular dynamics simulations revealed the binding mode and affinity between BI-4020 and the EGFR mutants. This study identified potential therapeutic strategies using new-generation macrocyclic EGFR inhibitor to overcome the emerging resistance mutants. Citation Format: Ryohei Katayama, Ken Uchibori, Makoto Nishio, Naoya Fujita. A macrocyclic kinase inhibitor overcomes the compound mutations that cause the resistance to all approved EGFR inhibitors in EGFR-positive lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4651.

Chronic Myeloid Leukemia Presenting with Megakaryocytic Blast Crisis: a Case Report with a Favorable Outcome

Chronic myeloid leukemia (CML) belongs to the chronic myeloproliferative neoplasms, a clonal disorder of pluripotent hematopoietic stem cells, arising because of a hallmark genetic anomaly- reciprocal translocation between chromosomes 9 and 22, known as the Philadelphia chromosome, which generates the BCR::ABL fusion gene. The disease can have a biphasic/triphasic evolution, comprised of an accelerated phase and a terminal blast phase (usually biphasic when blastic phase is lymphoid in nature). Most cases present during the chronic phase of the disease, with only 10% of cases presenting in the blast phase. Megakaryocytic blast crisis is an unusual form of presentation, accounting for less than 3% of cases and it commonly carries an unfavorable prognosis compared to the classic myeloid, lymphoid or even mixed lineage blast crises. Aim: To bring forth an unusual case of CML presenting with megakaryocytic blast crisis and the therapeutic challenges associated with this particular presentation, unexpectedly ending with a favorable outcome. Matherial and methods: we hereby present the case of a 50 year old female patient, diagnosed in 2015 with CML- megakaryocytic blast crisis, who underwent treatment with a tyrosine kinase inhibitor (TKI)- imatinib and intensive chemotherapy regimen 3+7, without satisfactory response. A second induction chemotherapy regimen followed (MEC), alltogether with the same TKI and cytoreductive treatment (hydroxyurea/HU), but still without achieving a complete remission. Thus, the TKI was switched to dasatinib and complete normalisation of blood counts followed (including thrombocytes). However, there was still inadequate molecular and cytogenetic response. Therefore, TKI dosage was increased and after 4 months the patient achieved complete remission, with no detectable BCR::ABL transcript. She spent 9 years on the same TKI until important pleural effusion developed, which required switching to another 2nd generation TKI- bosutinib, based on the patients age, genetic profile and commorbidities. She is still in complete remission without detectable BCR::ABL transcript/Philadelphia chromosome to this day (2024). Results and conclusions: this is the case of a rare presentation of CMLmegakaryocytic blast crisis, who underwent both TKI and intensive chemotherapy induction regimens, marked by a favorable unexpected outcome.

Abstract 6318: Development of an anti-PD-L1 CAR T therapeutic treating advanced gastric cancer with enhanced efficacy and lower off-tumor activities

Abstract Purpose: Driven by a relentless unmet need with unsatisfied cancer therapy, we aimed to develop chimeric antigen receptor T-cell (CAR-T) therapy for targeting advanced gastric cancer (AGC) expressing PD-L1. We constructed a 2nd generation CAR lentiviral vector expressing a unique anti-PD-L1 scFv with moderate affinity (VaxCAR001) in order to compare scFvs derived from Avelumab or Atezolizumab. This project focused on the efficacy and safety of PD-L1 CAR-Ts, which are affected by their binding affinity and dissociation with the target protein, PD-L1 during the treatment period. Methods and Results: The findings of this study revealed significant differences among PD-L1-targeting CAR-T cells with different affinities and dissociations. VaxCAR001 cells, showing intermediate affinity, demonstrated a higher CD8 ratio (up to 78% compared to 58% in Avelumab-derived and 45% in Atezolizumab-derived CAR-T), reduced expression of inhibitory markers including Tim-3, LAG-3, and TIGIT, (8%, 39% and 13% in VaxCAR001 vs 35%, 55% and 42 % in Avelumab, 19%, 51% and 34% in Atezolizumab scFv-derived CAR-T, respectively) and more sustained cytotoxicity. Notably, it maintained strong cytotoxicity even one month after production, while the Avelumab or Atezolizumab ScFv-derived CAR-T cells did not. In multicellular spheroid 3D co-culture with CAR T cells, VaxCAR001 exhibited superior infiltration into the core of spheroid and induced higher CAR-T-mediated killing for 3D tumor spheroids as indicated by LDH assay (68%, 800% increased than Avelumab or Atezolizumab scFv-derived CAR-T). Moreover, in an MSI-high AGC mouse model, treatment with VaxCAR001 cells resulted in a significant reduction in tumor size and prolonged survival until the end of the study period. On the other hand, treatment with the Avelumab or Atezolizumab scFv-derived CAR-T cells led to high mortality, with most mice succumbing within three weeks after CAR-T cell injection, suggesting an on-target off-tumor effect. Conclusion: This study demonstrates that VaxCAR001 cells bearing intermediate affinity and high dissociation rate for target PD-L1 showed the superior killing of PD-L expressing tumor cells in vitro and in vivo and manifested better safety profiles, as compared to CAR-T cells bearing scFv derived from commercial anti-PD-L1 targeting monoclonal antibodies such as avelumab and atezolizumab. VaxCAR001 revealed extended cytotoxicity, enhanced infiltration into tumor spheroids, and greater tumor size reduction in in vivo models. Importantly, it also demonstrated superior safety profiles, as evidenced by reduced expression of inhibitory markers and fewer off-tumor adverse effects. These findings would accelerate the clinical stage developments as an effective and safer treatment strategy against AGC. Citation Format: Minwoo Baek, Jae-In Yu, Eun-Ji Choi, Yeojin Lim, Jiwon Yang, Se-Ryeong Choi, Yoonjoo Choi, Mihwa Kim, Joon Haeng Rhee, Je-Jung Lee. Development of an anti-PD-L1 CAR T therapeutic treating advanced gastric cancer with enhanced efficacy and lower off-tumor activities [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 6318.

Abstract 4018: An innovative in vivo model for CAR-T cell therapy development: Tolerability and efficacy evaluation of CD19-targeting CAR-T cells on human lymphoma using the chicken CAM assay

Abstract BACKGROUND Chimeric antigen receptor (CAR) T cell therapy is a revolutionary approach in cellular immunotherapy, where a patient’s T cells are genetically engineered to recognize and attack cancer cells. In the last decade, several CAR-T cell therapies have been approved by the FDA. However, this encouraging outcome was obtained only with certain blood cancers, like B cell leukemias, lymphomas, etc. Many research/clinical trials are now underway worldwide to extend the CAR T-cell therapy benefits to a large spectrum of cancers, using rodents. This model presents several drawbacks, such as the immunodeficiency of humanized models, ethical constraints, time, and cost. An alternative, pertinent, 3Rs-compliant in vivo model for developing CAR-T therapy is urgently needed. Xenografts on the chicken embryo’s ChorioAllantoic Membrane (CAM) have proven extremely valuable for in vivo cancerology studies. In this work, we evaluated in vivo the anti-tumor efficacy of CD19-targeting CAR-T cells on human lymphoma using Inovotion’s CAM assay. METHODS Human lymphoma Raji cells (ATCC) were grafted on the CAM on embryonic development day (EDD) 9. A 2nd generation of CD19-targeting CAR-T cells (CAR-T-(CD3/4-1BB)) developed by Allogenica (France) were co-injected at EDD9 at E/T (effector CAR-T cell vs target Raji cell) ratio 1:1. One group was injected with CAR-T (CD3/4-1BB) only once at EDD9 and a second group received a first injection of CAR-T (CD3/4-1BB) at EDD9 and a second injection at EDD13. Embryonic viability was checked daily. On EDD18. The efficacy of CAR-T therapy was evaluated basing on tumor weight, metastatic invasion in the lower CAM and CAR-T infiltration in tumors detected by qPCR. Activation of the chicken embryo immune system was detected as well. RESULTS The treatment with CAR-T-(CD3/4-1BB) cells was well tolerated in ovo, no significant embryo mortality/abnormality was observed. CAR-T-(CD3/4-1BB) cells prevented tumor formation (tumor weight was 88.67% to 90.62% lower than control, p < 0.001); and prevented metastasis formation in the lower CAM (86.88% to 81.43% lower than control, p < 0.001). CONCLUSION Different to in vitro models and immunodeficient murine models, chicken embryo possesses a progressively mature immune system. Our results show that CAR-T-(CD3/4-1BB) cells can prevent tumor formation and metastatic invasion when co-injected with tumor cells. Inovotion’s CAM assay is a viable alternative in vivo model for developing CAR-T cell therapy on a large spectrum of cancers. It is fully 3Rs-compliant. Citation Format: Yan Wang, Xavier Rousset, Chloé Prunier, Emilien Dosda, Alejandra Gutierrez-Guerrero, Pauline Abrial, Inna Menkova, Jean Viallet. An innovative in vivo model for CAR-T cell therapy development: Tolerability and efficacy evaluation of CD19-targeting CAR-T cells on human lymphoma using the chicken CAM assay [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 4018.

UCOMB-real life data: treatment strategies for chronic urticaria patients with comorbidities

Background There is a lack of real-life safety data on treatment options for chronic urticaria in the presence of comedication and comorbidities. Methods We present a single-center UCARE pilot study of 212 outpatients with chronic urticaria. Patients were divided into three groups according to different CU therapies according to international guidelines. Results Of 212 patients, 108 (mean age 48.9 years, 71.3% female) had 59 comorbidities, including cardiovascular, autoimmune and malignant diseases. Patients were followed for a mean of 24.6 months (SD ± 21.3). Urticaria therapies were divided into three groups: A: 105 (97.2%) with omalizumab and 2nd generation antihistamines), B: 16 patients (14.8%): dual therapy with antihistamines and cyclosporine in 10 (9.3%), montelukast in five (4. 6%), dapsone in four (3.7%), hydroxychloroquine in one patient (0.9%), C: 12 (11.1%) patients received a third drug for 4.9 months (SD ± 3.2) and one quadruple therapy (2.1 months). 10 out of 12 (83.3%) patients received montelukast, two (16.7%) cyclosporine, two (16.7%) dapsone and one (8.3%) hydroxychloroquine as a third drug for chronic urticaria. Conclusions Combining treatment modalities for chronic urticaria and comorbidities are available and feasible with a good safety profile.

Blind deconvolution decreases requirements on temporal resolution of DCE-MRI: Application to 2nd generation pharmacokinetic modeling

PurposeDynamic Contrast-Enhanced (DCE) MRI with 2nd generation pharmacokinetic models provides estimates of plasma flow and permeability surface-area product in contrast to the broadly used 1st generation models (e.g. the Tofts models). However, the use of 2nd generation models requires higher frequency with which the dynamic images are acquired (around 1.5 s per image). Blind deconvolution can decrease the demands on temporal resolution as shown previously for one of the 1st generation models. Here, the temporal-resolution requirements achievable for blind deconvolution with a 2nd generation model are studied. MethodsThe 2nd generation model is formulated as the distributed-capillary adiabatic-tissue-homogeneity (DCATH) model. Blind deconvolution is based on Parker's model of the arterial input function. The accuracy and precision of the estimated arterial input functions and the perfusion parameters is evaluated on synthetic and real clinical datasets with different levels of the temporal resolution. ResultsThe estimated arterial input functions remained unchanged from their reference high-temporal-resolution estimates (obtained with the sampling interval around 1 s) when increasing the sampling interval up to about 5 s for synthetic data and up to 3.6–4.8 s for real data. Further increasing of the sampling intervals led to systematic distortions, such as lowering and broadening of the 1st pass peak. The resulting perfusion-parameter estimation error was below 10% for the sampling intervals up to 3 s (synthetic data), in line with the real data perfusion-parameter boxplots which remained unchanged up to the sampling interval 3.6 s. ConclusionWe show that use of blind deconvolution decreases the demands on temporal resolution in DCE-MRI from about 1.5 s (in case of measured arterial input functions) to 3–4 s. This can be exploited in increased spatial resolution or larger organ coverage.

Past, present, and future of microbiome-based therapies.

Technological advances in studying the human microbiome in depth have enabled the identification of microbial signatures associated with health and disease. This confirms the crucial role of microbiota in maintaining homeostasis and the host health status. Nowadays, there are several ways to modulate the microbiota composition to effectively improve host health; therefore, the development of therapeutic treatments based on the gut microbiota is experiencing rapid growth. In this review, we summarize the influence of the gut microbiota on the development of infectious disease and cancer, which are two of the main targets of microbiome-based therapies currently being developed. We analyze the two-way interaction between the gut microbiota and traditional drugs in order to emphasize the influence of gut microbial composition on drug effectivity and treatment response. We explore the different strategies currently available for modulating this ecosystem to our benefit, ranging from 1st generation intervention strategies to more complex 2nd generation microbiome-based therapies and their regulatory framework. Lastly, we finish with a quick overview of what we believe is the future of these strategies, that is 3rd generation microbiome-based therapies developed with the use of artificial intelligence (AI) algorithms.

Does progress in microfracture techniques necessarily translate into clinical effectiveness?

Multitudinous advancements have been made to the traditional microfracture (MFx) technique, which have involved delivery of various acellular 2nd generation MFx and cellular MFx-III components to the area of cartilage defect. The relative benefits and pitfalls of these diverse modifications of MFx technique are still not widely understood. To comparatively analyze the functional, radiological, and histological outcomes, and complications of various generations of MFx available for the treatment of cartilage defects. A systematic review was performed using PubMed, EMBASE, Web of Science, Cochrane, and Scopus. Patients of any age and sex with cartilage defects undergoing any form of MFx were considered for analysis. We included only randomized controlled trials (RCTs) reporting functional, radiological, histological outcomes or complications of various generations of MFx for the management of cartilage defects. Network meta-analysis (NMA) was conducted in Stata and Cochrane's Confidence in NMA approach was utilized for appraisal of evidence. Forty-four RCTs were included in the analysis with patients of mean age of 39.40 (± 9.46) years. Upon comparing the results of the other generations with MFX-I as a constant comparator, we noted a trend towards better pain control and functional outcome (KOOS, IKDC, and Cincinnati scores) at the end of 1-, 2-, and 5-year time points with MFx-III, although the differences were not statistically significant (P > 0.05). We also noted statistically significant Magnetic resonance observation of cartilage repair tissue score in the higher generations of microfracture (weighted mean difference: 17.44, 95% confidence interval: 0.72, 34.16, P = 0.025; without significant heterogeneity) at 1 year. However, the difference was not maintained at 2 years. There was a trend towards better defect filling on MRI with the second and third generation MFx, although the difference was not statistically significant (P > 0.05). The higher generations of traditional MFx technique utilizing acellular and cellular components to augment its potential in the management of cartilage defects has shown only marginal improvement in the clinical and radiological outcomes.

Do immigrants support environmentalism? – Immigrants’ environmental socialization in the United States

As globalisation continues, heated debates over immigrants’ environmental behaviours in the U.S. have been prominent in recent years. While research has generally focused on the association between political affiliation and pro-environmental values, only a few studies have specifically explored immigrants’ pro-environmental values in the U.S. Utilizing a pooled dataset spanning 3 years (2017, 2020, and 2022) from the Cooperative Election Study and employing a multilevel model, we empirically tested three hypotheses regarding immigrants’ pro-environmental attitudes: the globalization hypothesis, the prosperity hypothesis, and the political socialization hypothesis. Our results supported all three hypotheses. On average, immigrants exhibited higher environmental scores than the native-born population. In addition, we identified generational differences, with the 1st generation immigrants showing the highest environmental scores, while the 2nd generation immigrants fell between the 1st generation and the native-born. Compared to the native-born population, party affiliation played a smaller role in immigrants’ formation of pro-environmental values. Democratic immigrants tended to be less pro-environment than democratic locals, while Republican immigrants were more pro-environment than Republican locals.

Allogeneic stem cell transplantation is still a highly curative therapy in adults with philadelphia chromosome-positive acute lymphoblastic leukaemia.

The application of tyrosine kinase inhibitors (TKIs) and novel immunotherapies has improved outcomes in patients with Ph + acute lymphoblastic leukaemia (ALL), and the issue of whether there is still a need for stem cell transplantation has become controversial. We performed a retrospective study to explore whether stem cell transplantation still held a place in patients with Ph + ALL if only imatinib and 2nd generation TKIs are available and affordable. A total of 292 patients were included. The median age was 38 years [range 14-64, IQR 28-48]. Patients receiving transplants (n = 216) had better rates of 4-year disease-free survival (DFS, 68% vs. 24%, P < .0001) and overall survival (OS, 72% vs. 47%, P < .0001) than those receiving continuous TKIs plus chemotherapy (TKI-chemo) (n = 76). In the multivariate analysis, male sex, WBC count ≥ 95 × 109/L and PLT count ≤ 154 × 109/L at diagnosis were significantly associated with poorer outcomes, and transplantation was significantly associated with favourable DFS and OS. In addition, the transplant outcomes were superior in any subgroup according to the number of risk variables. Furthermore, propensity score matching (PSM) analyses showed similar findings in the whole cohort and in age- and BCR-ABL1 level-based subgroups after the first or second consolidation. In conclusion, transplantation as a one-time procedure for adults with Ph + ALL patients remains important in countries lacking accessibility to third-generation TKIs or immunotherapies, regardless of the depth of the molecular response.

Impact of RON on a heavily downsized boosted SI engine using 2nd generation biofuel – A comprehensive experimental analysis

Sustainable energy solutions are paramount in the urgent global drive against climate change, especially in transportation. This research focuses on second-generation biogasolines and their potential in the context of decarbonisation. Two biogasolines, 99 RON E20 and 95 RON E20, were rigorously tested in a downsized single-cylinder engine. Their performance, combustion, and emissions were compared against the conventional fossil fuel, 95 RON E10, under varying engine loads. Additionally, a comprehensive injection parameter sweep was conducted for both biofuels at low and high loads, shedding light on their unique operational characteristics and operational regimes. This research significantly enhances our knowledge about the potential of these new biofuels and their implications for a more sustainable energy future. The findings of the experiments demonstrate no substantial difference between the tested biofuels and fossil fuels. Biofuel with a higher octane number provides more knock resistance than fossil fuel, resulting in increased thermal efficiency due to spark advance ability. However, more significant hydrocarbon emissions were detected for biofuels than fossil fuels due to more extensive aromatic content. Both biofuels have stable combustion in low and high-load operations under varying injection pressures and injection start times. 99 Bio E20 has a wider operational range than 95 Bio E20. However, due to very high HC emissions, especially at high-load operations, an early injection start with more significant injection pressure is not recommended for biofuel. From a broader perspective, both biofuels exhibit the promising potential to serve as drop-in replacements in spark ignition engines.

Perceived discrimination in health care in Germany– results of a population survey

BackgroundIt has consistently been shown that perceived discrimination is associated with adverse health outcomes. Despite this uncontested relevance, there is a lack of research on the experiences of discrimination in health care. Therefore, the following research questions were addressed: (1) How often do people in Germany report having been discriminated in health care due to different reasons? (2) Which socio-demographic groups are most afflicted by perceived discrimination in health care?MethodsAnalyses are based on a cross-sectional online survey conducted in Germany. An adult population sample was randomly drawn from a panel which was recruited offline (N = 2,201). Respondents were asked whether they have ever been discriminated in health care due to the following reasons: age, sex/gender, racism (i.e. migration history, religion, language problems, colour of skin), health issues or disability (i.e. overweight, mental illness/addiction, disability), socio-economic status (SES, i.e. income, education, occupation).Results26.6% of the respondents reported discrimination experiences. Perceived discrimination due to health issues or disability was most frequent (15%), followed by age (9%) and SES (8.9%). Discrimination due to racism and sex/gender was less frequently reported (4.1% and 2.5%). Younger age groups, women, and 2nd generation migrants as well as respondents with low income and low education were more likely to report any kind of discrimination in health care. Two groups were found to be at special risk for reporting discrimination in health care across different reasons: women and younger age groups. Discrimination due to racism was more prevalent among respondents who have immigrated themselves than those who were born in Germany but whose parents have immigrated. Discrimination due to SES was significantly associated with (low) income but not with education.ConclusionsMore than a quarter of the adult population in Germany reported experiences of discrimination in health care. Such experiences were more frequent among lower SES groups, migrants, women, and younger people. Results underline the necessity of interventions to reduce the magnitude and consequences of discrimination in health care. Future studies should apply an intersectional approach to consider interactions between social inequality indicators regarding discrimination and to identify risk groups that are potentially afflicted by multiple discrimination.

Kraft (Nano)Lignin as Reactive Additive in Epoxy Polymer Bio-Composites.

The demand for high-performance bio-based materials towards achieving more sustainable manufacturing and circular economy models is growing significantly. Kraft lignin (KL) is an abundant and highly functional aromatic/phenolic biopolymer, being the main side product of the pulp and paper industry, as well as of the more recent 2nd generation biorefineries. In this study, KL was incorporated into a glassy epoxy system based on the diglycidyl ether of bisphenol A (DGEBA) and an amine curing agent (Jeffamine D-230), being utilized as partial replacement of the curing agent and the DGEBA prepolymer or as a reactive additive. A D-230 replacement by pristine (unmodified) KL of up to 14 wt.% was achieved while KL-epoxy composites with up to 30 wt.% KL exhibited similar thermo-mechanical properties and substantially enhanced antioxidant properties compared to the neat epoxy polymer. Additionally, the effect of the KL particle size was investigated. Ball-milled kraft lignin (BMKL, 10 μm) and nano-lignin (NLH, 220 nm) were, respectively, obtained after ball milling and ultrasonication and were studied as additives in the same epoxy system. Significantly improved dispersion and thermo-mechanical properties were obtained, mainly with nano-lignin, which exhibited fully transparent lignin-epoxy composites with higher tensile strength, storage modulus and glass transition temperature, even at 30 wt.% loadings. Lastly, KL lignin was glycidylized (GKL) and utilized as a bio-based epoxy prepolymer, achieving up to 38 wt.% replacement of fossil-based DGEBA. The GKL composites exhibited improved thermo-mechanical properties and transparency. All lignins were extensively characterized using NMR, TGA, GPC, and DLS techniques to correlate and justify the epoxy polymer characterization results.

Comparison of two parathyroid hormone (pth) assay methods in the monitoring of black african hemodialysis patients in the ivory coast (Maglumi® vs Vidas Biomerieux® Kits)

Parathyroid hormone (PTH) testing is recommended once or twice a year to prevent bone complications resulting from chronic kidney disease. However, its interpretation is subject to many factors, including the generation of the PTH test used. For this reason, it is recommended that each laboratory establish its own PTH Upper Limit of Normality (ULN). In order to guarantee optimal management of hemodialysis patients in the public sector in Côte d'Ivoire (West Africa), it proved important to compare the diagnosis of hyperparathyroidism made using two platforms used in the laboratory in charge of monitoring these patients: PTH VIDAS® (3rd Generation) and PTH MAGLUMI® (2nd Generation). This was a cross-sectional study of 65 haemodialysis patients in Abidjan. PTH determinations were performed simultaneously on VIDAS® and MAGLUMI® platforms. The results obtained were interpreted on the basis of PTH threshold values pre-established in the laboratory. 86.15% of patients had concordant diagnoses on both platforms, compared with 13.85% with discordant diagnoses. Statistical analysis of these results showed a non-statistically significant difference.: The limitation posed by differences in PTH concentration given by different platforms can be circumvented by using PTH ULN stablished for each platform for a given population. Conclusion: The diagnoses made by the two platforms are thus superimposable using their respective PTH ULN.

Analysis of the effect of baseline detection and early clearance of ct-DNA, on survival outcomes among patients with advanced EGFR-mutant non-small cell lung cancer.

To determine if circulating tumor DNA (ct-DNA) dynamics of epidermal growth factor receptor (EGFR) mutation in plasma can identify asubset of patients with EGFR-mutant (EGFR- m) non-small cell lung cancer (NSCLC) with inferior survival outcomes, we analyzed and compared survival outcomes among patients with and without baseline presence and early clearance of EGFR ct-DNA in plasma. For 66patients newly dia-gnosed with EGFR- m NSCLC, plasma samples were collected at baseline and 1st response assessment at 12-24weeks for extraction of ct-DNA. Estimation of ct-DNA (EGFR exons 18, 19, 20and 21) was done using droplet digital polymerase chain reaction (dd-PCR) on the QX200ddPCR system (BioRad, USA). Patients with detectable EGFR ct-DNA at baseline (sample 1), with either undetectable or persistent detectable ct-DNA in sample 2were classified as clearers and non-clearers, respectively. Fifty-three patients received 1st/ 2nd generation EGFR tyrosine kinase inhibitors (TKIs) and 13received either 3rd generation TKI (osimertinib) or chemotherapy plus gefitinib. The baseline ct-DNA-positive group had more patients with extra thoracic disease (60.4vs. 48.5%). For the entire cohort, there was no difference in median progression-free survival (PFS) among baseline ct-DNA-negative (13.57months) vs. ct-DNA-positive patients (12.32months) (HR 0.74). There was asignificant improvement of PFS among early ct-DNA clearers vs. non-clearers (12.32vs. 9.92months; HR 0.57). For those treated with 1st/ 2nd generation EGFR TKIs, this improvement in median PFS among early ct-DNA clearers vs. non-clearers was more apparent (11.76vs. 6.8months; HR 0.34). Baseline detection of the presence of ct-DNA of EGFR mutation in plasma was not predictive of first-line PFS, but is associated with extra thoracic disease. Patients with EGFR mutation and persistence of ct-DNA at first follow-up have worse PFS and overall survival (OS) in comparison to those clearing the same in plasma, especially among those treated with 1st/ 2nd generation EGFR TKIs.

Diabetes quality of life, Chemical Intolerance, and generational status in a Latino sample: an RRNeT study

ObjectivesThe impact of diabetes on quality of life (QoL) includes deficits in physical functioning and emotional and mental health. Individuals with diabetes can experience significant distress related to diabetes management and burden. Comorbid conditions can contribute to QoL among individuals with diabetes. Chemical Intolerance (CI) has received minimal attention in the context of diabetes. CI is characterized by multi-systemic symptoms initiated by a one-time high dose or persistent low-dose exposure to environmental toxins. Latinos experience diabetes distress and are exposed to environmental factors at disproportionate rates. This study sought to investigate generational differences in diabetes QoL and CI in a Latino sample across generational status.MethodsTo assess the modifying effect of CI between QoL and generations, the analysis was stratified by those with and without CI.ResultsCompared to 2nd generation Latinos, Latino immigrants were more likely to indicate that without diabetes, their life would be better across several QoL domains. Latino immigrants had a lower A1C and BMI than2nd generation Latinos. Additionally, they were less likely to have scores indicative of CI than U.S.-born Latinos. QoL varied by generational status and suggested a greater emphasis on family and social relationships among immigrants than U.S.-born Latinos. There were no differences in health services measures across generations.ConclusionsThis study highlights generational differences in the impact of diabetes on QoL. It is one of the few studies to seek to understand the impact of CI on QoL in the context of a chronic condition such as type 2 diabetes.

Towards the production of net-negative greenhouse gas emission bio-based plastics from 2nd and 3rd generation feedstocks

Here, we show production pathways for greenhouse gas (GHG)-negative bio-based plastics from 2nd and 3rd generation feedstocks. We focus on bio-based plastics that are technically capable of replacing 80% of the global plastic market. By presenting life cycle inventories and discussing GHG-emissions hotspots, this work will inform stakeholders along the plastic supply chain of the necessary steps to achieving net-zero emissions by 2050, and potentially, how to drive net-uptake. This work is of critical importance given the overwhelming mass of plastic produced annually and the resulting CO2 emissions. To conduct this assessment, we derive life cycle inventories for nine different bio-based plastics and address the impact of methodological choices, such as allocation method, on the resulting 100a global warming potential (GWP). Our findings show that resources used and processing methods implemented have significant effects on the potential for us to derive carbon-negative plastics. Furthermore, we find that environmental impact quantification methods greatly influence the perceived GWP of such processes. For example, economic and mass allocation methods resulted in an apparent increase in GWP of up to 39% and 166%, respectively, compared to no allocation for bio-based plastics made from 2nd generation crops, whereas mass allocation resulted in the lowest GWP for bio-based plastics made from 1st generation crops. In considering environmental impact hotspots, our findings show that decarbonization of thermal energy and electricity, reduced use of ammonia-based fertilizer, renewable hydrogen production, use of bio-based alternatives for petrochemicals and plasticizers, enzyme production pathways from 2nd generation crops, and more efficient biomass conversion processes to reduce feedstock inputs may be critical steps in creating GHG-negative bio-based plastics in the future.

Quantification of blood glial fibrillary acidic protein using a second-generation microfluidic assay. Validation and comparative analysis with two established assays.

Increased levels of glial fibrillary acidic protein (GFAP) in blood have been identified as a valuable biomarker for some neurological disorders, such as Alzheimer's disease and multiple sclerosis. However, most blood GFAP quantifications so far were performed using the same bead-based assay, and to date a routine clinical application is lacking. In this study, we validated a novel second-generation (2nd gen) Ella assay to quantify serum GFAP. Furthermore, we compared its performance with a bead-based single molecule array (Simoa) and a homemade GFAP assay in a clinical cohort of neurological diseases, including 210 patients. Validation experiments resulted in an intra-assay variation of 10 %, an inter-assay of 12 %, a limit of detection of 0.9 pg/mL, a lower limit of quantification of 2.8 pg/mL, andless than 20 % variation in serum samples exposed toupto five freeze-thaw cycles, 120 h at 4 °C and room temperature. Measurement of the clinical cohort using allassaysrevealed the same pattern of GFAP distribution inthedifferent diagnostic groups. Moreover, we observed a strong correlation between the 2nd gen Ella and Simoa (r=0.91 (95 % CI: 0.88-0.93), p<0.0001) and the homemade immunoassay (r=0.77 (95 % CI: 0.70-0.82), p<0.0001). Our results demonstrate a high reliability, precision and reproducibility of the 2nd gen Ella assay. Although a higher assay sensitivity for Simoa was observed, the new microfluidic assay might have the potential to be used for GFAP analysis in daily clinical workups due to its robustness and ease of use.