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Follicular lymphoma (FL) is a B-cell malignancy with a complex tumor microenvironment that is rich in nonmalignant immune cells. We applied single-cell RNA sequencing to characterize the diverse tumor and immune cell populations of FL and identified major phenotypic subsets of FL T cells, including a cytotoxic CD4 T-cell population. We characterized four major FL subtypes with differential representation or relative depletion of distinct T-cell subsets. By integrating exome sequencing, we observed that somatic mutations are associated with, but not definitive for, reduced MHC expression on FL cells. In turn, expression of MHCII genes by FL cells was associated with significant differences in the proportions and targetable immunophenotypic characteristics of T cells. This provides a classification framework of the FL microenvironment in association with FL genotypes and MHC expression, and informs different potential immunotherapeutic strategies based upon tumor cell MHCII expression. Significance: We have characterized the FL-infiltrating T cells, identified cytotoxic CD4 T cells as an important component that is associated with tumor cell–intrinsic characteristics, and identified sets of targetable immune checkpoints on T cells that differed from FLs with normal versus low MHC expression. See related commentary by Melnick, p. 374. This article is highlighted in the In This Issue feature, p. 369

Supplementary Figure from Follicular Lymphoma Microenvironment Characteristics Associated with Tumor Cell Mutations and MHC Class II Expression

Supplementary Data from Follicular Lymphoma Microenvironment Characteristics Associated with Tumor Cell Mutations and MHC Class II Expression

Follicular lymphoma (FL) is a B-cell malignancy with a complex tumor microenvironment that is rich in nonmalignant immune cells. We applied single-cell RNA sequencing to characterize the diverse tumor and immune cell populations of FL and identified major phenotypic subsets of FL T cells, including a cytotoxic CD4 T-cell population. We characterized four major FL subtypes with differential representation or relative depletion of distinct T-cell subsets. By integrating exome sequencing, we observed that somatic mutations are associated with, but not definitive for, reduced MHC expression on FL cells. In turn, expression of MHCII genes by FL cells was associated with significant differences in the proportions and targetable immunophenotypic characteristics of T cells. This provides a classification framework of the FL microenvironment in association with FL genotypes and MHC expression, and informs different potential immunotherapeutic strategies based upon tumor cell MHCII expression.Significance:We have characterized the FL-infiltrating T cells, identified cytotoxic CD4 T cells as an important component that is associated with tumor cell–intrinsic characteristics, and identified sets of targetable immune checkpoints on T cells that differed from FLs with normal versus low MHC expression.See related commentary by Melnick, p. 374.This article is highlighted in the In This Issue feature, p. 369

The historical lack of preclinical models reflecting the genetic heterogeneity of multiple myeloma (MM) hampers the advance of therapeutic discoveries. To circumvent this limitation, we screened mice engineered to carry eight MM lesions (NF-κB, KRAS, MYC, TP53, BCL2, cyclin D1, MMSET/NSD2 and c-MAF) combinatorially activated in B lymphocytes following T cell-driven immunization. Fifteen genetically diverse models developed bone marrow (BM) tumors fulfilling MM pathogenesis. Integrative analyses of ∼500 mice and ∼1,000 patients revealed a common MAPK–MYC genetic pathway that accelerated time to progression from precursor states across genetically heterogeneous MM. MYC-dependent time to progression conditioned immune evasion mechanisms that remodeled the BM microenvironment differently. Rapid MYC-driven progressors exhibited a high number of activated/exhausted CD8+ T cells with reduced immunosuppressive regulatory T (Treg) cells, while late MYC acquisition in slow progressors was associated with lower CD8+ T cell infiltration and more abundant Treg cells. Single-cell transcriptomics and functional assays defined a high ratio of CD8+ T cells versus Treg cells as a predictor of response to immune checkpoint blockade (ICB). In clinical series, high CD8+ T/Treg cell ratios underlie early progression in untreated smoldering MM, and correlated with early relapse in newly diagnosed patients with MM under Len/Dex therapy. In ICB-refractory MM models, increasing CD8+ T cell cytotoxicity or depleting Treg cells reversed immunotherapy resistance and yielded prolonged MM control. Our experimental models enable the correlation of MM genetic and immunological traits with preclinical therapy responses, which may inform the next-generation immunotherapy trials.

SUMMARYReference atlases, molecular and spatial maps of mammalian tissues, are critical resources for discovery efforts and translational research. Their utility is dependent on operationalizing the resulting data by identifying cell types, histological patterns, and predictive biomarkers underlying health and disease. The human lymph node (LN) offers a compelling use case because of its importance in immunity, structural and cellular diversity, and neoplastic involvement. One hematological malignancy, follicular lymphoma (FL), evolves from developmentally blocked germinal center B cells residing in and trafficking through these tissues. To promote survival and immune escape, tumor B cells undergo significant genetic changes and extensively remodel the lymphoid microenvironment. Here, we present an integrated portrait of healthy and FL LNs using multiple genomic and advanced imaging technologies. By leveraging the strengths of each platform, we identified several tumor-specific features and microenvironmental patterns enriched in individuals who experience early relapse, the most high-risk of FL patients.

Recent advances in immunotherapy demonstrate the need to further understand the characteristics of an individual cancer patient’s immune system and how it influences responses to cancer treatment. Here, we developed an immunoprofiling platform to evaluate the features in the blood of cancer patients to test the hypothesis that peripheral immune cell heterogeneity could be used to stratify these patients into different categories or immunotypes to monitor disease progression and treatment response. To that end, we established a unique diagnostic immunoprofiling assay and analytical framework based on the analysis of leukocytes in the peripheral blood using multiparameter flow cytometry. Supervised manual gating of flow cytometry data from a cohort of 50 healthy donors identified 415 cell types and immune activation states that were used to train and later independently validate machine learning models to automatically identify immune cell subsets from raw cytometry data. By applying this tool to peripheral blood samples from a mixed cohort of 299 healthy donors and 323 cancer patients, we developed a machine-learning classification model that can differentiate between these two groups with 93% accuracy. This model was further refined using spectral clustering with bootstrapping, revealing 5 clusters, or immunotypes, characterized by specific physiological immune profiles: (1) Myeloid-derived suppressor/NK cell, (2) Terminally-differentiated CD8+ T cells, (3) Mixed CD4+ T helper cells, (4) CD4+ Th1 & CD8+ T cell memory, and (5) Naive T and B lymphocytes. Interestingly, very few healthy donors could be found in clusters 1 and 2 but were assigned most frequently to cluster 5. Matched RNA-seq was used to further validate these profiles using the cellular deconvolution algorithm, Kassandra, and differential gene expression analysis revealed immunotype-specific signatures that are consistent with immune response potential. Patients in the terminally-differentiated CD8+ T cell cluster had a narrower range of HLA-types than the other clusters, and TCR repertoire analysis indicated significantly increased clonality and reduced clonotype diversity. Within this cluster there was a high degree of overlap between TCR sequences in the peripheral blood and the tumor, indicating a relationship between peripheral blood immunotype and tumor infiltration. Altogether, the establishment of these immunotypes using peripheral blood immunoprofiling represents a promising signature that can be used to identify and stratify cancer patients that will benefit from immune-based therapies. Citation Format: Daniiar Dyikanov, Iris Wang, Tatiana Vasileva, Polina Shpudeiko, Polina Turova, Arseniy A. Sokolov, Olga Golubeva, Evgenii Tikhonov, Anna Kamysheva, Ilya Krauz, Mary Abdou, Madison Chasse, Tori Conroy, Nicholas R. Merriam, Boris Shpak, Anastasia Radko, Anastasiia Kilina, Lira Nigmatullina, Linda Balabanian, Christopher J. Davitt, Alexander A. Ryabykh, Olga Kudryashova, Cagdas Tazearslan, Ravshan Ataullakhanov, Alexander Bagaev, Aleksandr Zaitsev, Nathan Fowler, Michael F. Goldberg. Comprehensive immunoprofiling of peripheral blood reveals five conserved immunotypes with implications for immunotherapy in cancer patients [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6664.

We present here the results of our investigations concerning the usage of «O2 alive» water for the pre-sowing seeds treatment and plant growth-promoting effect. It has been found that the pre-sowing seeds treatment with the «O2 alive» water results in the increase of the seed vigor and germinating ability of the seeds at about 80-100%, at the same time this procedure accelerates the onset of the most important phenophases of plant development and increases the ability of plant to appropriate a fertilizer added to soil. It has been shown that the simultaneous mineral fertilization («Zdraven» fertilizer) and the usage of the «O2 alive» water increases the productivity of the protected and open-ground plants by about 30-80% and improves the plants flexibility to the unfavorable conditions (drought, low temperatures).

Personalized cancer vaccines designed to target neoantigens represent a promising new treatment paradigm in oncology. In contrast to classical idiotype vaccines, we hypothesized that ‘polyvalent’ vaccines could be engineered for the personalized treatment of follicular lymphoma (FL) using neoantigen discovery by combined whole exome sequencing (WES) and RNA sequencing (RNA-Seq). Fifty-eight tumor samples from 57 patients with FL underwent WES and RNA-Seq. Somatic and B-cell clonotype neoantigens were predicted and filtered to identify high-quality neoantigens. B-cell clonality was determined by alignment of B-cell receptor (BCR) CDR3 regions from RNA-Seq data, grouping at the protein level, and comparison to the BCR repertoire of RNA-Seq data from healthy individuals. An average of 52 somatic mutations per patient (range: 2-172) were identified, and two or more (median: 15) high-quality neoantigens were predicted for 56 of 58 samples. The predicted neoantigen peptides were composed of missense mutations (76%), indels (9%), gene fusions (3%), and BCR sequences (11%). Building off of these preclinical analyses, we initiated a pilot clinical trial using personalized neoantigen vaccination combined with PD-1 blockade in patients with relapsed or refractory FL (#NCT03121677). Synthetic long peptide (SLP) vaccines were successfully synthesized for and administered to all four patients enrolled to date. Initial results demonstrate feasibility, safety, and potential immunologic and clinical responses. Our study suggests that a genomics-driven personalized cancer vaccine strategy is feasible for patients with FL, and this may overcome prior challenges in the field.

We investigated the microstructure of particles after ultrasonic treatment of aqueous suspensions of detonation synthesis nanodiamond (DND) and polycrystalline detonation nanodiamond (DALAN) powders. After processing and drying the powder, we measured the specific surface area (by the BET method), the sizes of microcrystallites, the degree of crystallinity and the parameters of the crystal lattice. The study of the phase composition and microstructure was carried out by X-ray diffraction analysis on an XRD-6000 diffractometer using CuKa radiation. The phase composition analysis was carried out using PDF 4+ databases, as well as the POWDER CELL 2.4 full profile analysis program. The particle size distribution of the obtained oil suspensions was measured using an OLYMPUS OMEC DC130 optical analyzer. It is shown that the size of microcrystallites, the degree of crystallinity of nanodiamond particles decreases with ultrasonic treatment. The specific surface area of DND particles decreased, while for DALAN it increased after the ultrasound. On the other hand, microstresses of the second kind increase significantly, which means that the excess surface energy stored in the particles also increases. Possible mechanisms of changes in the considered characteristics of nanodiamond particles under the action of ultrasound are discussed.

Disperse systems are widely used in technology (medicine, food science, oil refining, metallurgy, etc [...]

Human papillomavirus (HPV)-associated Head and Neck Squamous Cell Carcinoma (HPV+HNSCC) is now the most common HPV-associated malignancy in the United States. Current treatments can be associated with severe side-effects or lack of efficacy yet prognostic biomarkers are limited, slowing efforts to personalize treatment in HPV+HNSCC. Here, we describe the use of a transcriptomic-based analytical platform to analyze expression patterns of viral transcripts, the tumor microenvironment (TME), and viral genome integration, and associate these features with overall survival. Functional gene expression signatures were analyzed on publicly available HPV+HNSCC expression data (n=266). Unsupervised clustering analysis revealed 5 distinct and novel TME types across patients (immune-enriched non-fibrotic, immune-enriched fibrotic, fibrotic, immune-desert, immune-enriched luminal). These microenvironment subtypes were highly correlated with both overall survival and patient prognosis. Tumors with an immune-enriched microenvironment showed the highest survival rates, whereas fibrotic TME types were associated with poor survival (p < 0.05). Unsupervised clustering of a HPV+HNSCC cohort from The Cancer Genome Atlas (TCGA) (n=53), based on HPV transcript expression, revealed 4 HPV-related subtypes. Each subtype was enriched for distinct viral transcripts: E2/E5, E6/E7, E1/E4 and L1/L2. We then validated TME and HPV transcript-related classifications on an independent HPV+HNSCC cohort (n=132). Utilizing both viral transcript and TME subtypes, we found that the E2/E5 HPV subtype was associated with an immune-enriched TME and had a higher overall survival compared to the other subtypes. The E2/E5 subtype was also enriched for samples without HPV-genome integration, suggesting that HPV episomal DNA status and E2/E5 expression pattern may drive an inflamed microenvironment and improved prognosis. In contrast, E6/E7 subtype samples were associated with the fibrotic and depleted TME types, with lower values of T-cell and B-cell gene expression signatures and a lower survival rate. Both E1/E4 and L1/L2 subtypes were associated with the immune-enriched luminal TME types. These findings suggest that HPV-transcript expression patterns may drive modulation of the TME, and hence impact prognosis. Further validation of the relationships between viral gene expression, TME, and prognosis is warranted to understand if such subtypes could aid in the development of prognostic biomarkers for treatment selection. Citation Format: Daria Kiriy, Dmitry Tychinin, Nikita Kotlov, Olga Kudryashova, Anastasia Nikitina, Andrey Tyshevich, Naira Samarina, Ksenia Demina, Sandrine Degryse, Susan Raju Paul, Mark Poznansky, Krystle Lang Kuhs, James S. Lewis, Robert L. Ferris, Xiaowei Wang, Alexander Bagaev, Nathan Fowler, Lori Wirth, Daniel Faden. Viral transcript and tumor immune microenvironment-based transcriptomic profiling of HPV-associated head and neck cancer identifies subtypes associated with prognosis [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 3823.

589 Background: Spatiotemporal heterogeneity, paucity of actionable targets, and complexity of the tumor microenvironment (TME) are major barriers to therapeutic advances in pancreatic ductal adenocarcinoma (PDAC). We reconstructed the transcriptomic data from a heterogeneous cohort of PDAC patients (pts) to examine the TME and identify putative therapeutic strategies. Methods: Transcriptomic profiling and targeted gene sequencing data (Tempus) on primary or metastatic specimens from PDAC pts treated at the Medical College of Wisconsin (MCW) between 2015-2020 were analyzed. Mutation calling, expression analysis, cell type deconvolution from the transcriptome, and TME reconstruction were performed using BostonGene’s automated pipelines. Mann-Whitney U test and Fisher's exact test were used to assess statistical significance. Results: The cohort (N = 79) comprised of resectable (19%), borderline resectable (37%), locally advanced (24%) and metastatic (20%) PDAC pts. The most frequently used tumor sites for transcriptomic profiling were pancreas primary (59%), liver (16%), lung (10%) and peritoneum (10%). Four distinct subtypes were identified based on the BostonGene classification of the transcriptomic TME– Immune Enriched (IE; 14%), Fibrotic (F; 28%), Immune Enriched & Fibrotic (IEF; 36%), and Immune Depleted (ID; 22%). Analyses of the cellular composition of the TME subtypes with RNA-seq-based deconvolution showed that T-cell fractions (CD4, CD8) were higher in the IE/IEF subtypes compared to the F/ID subtypes (CD8 means: 6.4% vs 2.9%, p < 0.001; CD4 means: 15.1% vs. 7.6%, p < 0.001), while fibroblast content was higher in the F/IEF subtypes compared to the IE/ID subtypes (37.4% vs 18.4%; p < 0.001). KRAS wild-type (WT) tumors were enriched in the IEF subtype (58%), while KRAS mutated tumors comprised all four transcriptomic subtypes. Primary PDACs that underwent radiotherapy were significantly more enriched in fibroblasts compared to samples from the TCGA cohort that did not undergo radiotherapy (means: 30%(MCW) vs. 20% (TCGA), p < 0.001). Primary PDACs were enriched in the IEF subtype (46%), while liver and lung metastases were enriched in the ID (74%) and IE subtypes (70%), respectively. When pts were dichotomized to short (< 400 days) versus long (> 800 days) survivors, tumors from pts with longer survival demonstrated a trend towards enrichment in CD4/CD8 T cells and IE subtype that did not meet statistical significance. Conclusions: Lung metastases and KRAS WT PDACs harbor an immunogenic TME while liver metastases harbor an immune-cold TME, highlighting the biologic heterogeneity of PDAC. The efficacy of immunotherapeutic strategies in PDAC pts who demonstrate an IE/IEF transcriptomic subtype merits prospective evaluation. The four distinct subtypes identified by TME transcriptomic classification highlight the possibility of personalized immunotherapeutic strategies in PDAC.

This article dwells upon the additive manufacturing of high-energy materials (HEM) with regards to the problems of this technology’s development. This work is aimed at identifying and describing the main problems currently arising in the use of AM for nanostructured high-energy materials and gives an idea of the valuable opportunities that it provides in the hope of promoting further development in this area. Original approaches are proposed for solving one of the main problems in the production of nanostructured HEM—safety and viscosity reduction of the polymer-nanopowder system. Studies have shown an almost complete degree of deagglomeration of microencapsulated aluminum powders. Such powders have the potential to create new systems for safe 3D printing using high-energy materials.

Background: Follicular lymphoma (FL) shows marked variation in clinical course including spontaneous regression and histologic transformation (HT). Watchful waiting (W&W) is routinely applied to pts with newly diagnosed FL, but monitoring strategies are not standardized. Pts with progression within 1-2y of diagnosis have worse outcomes, but the biologic basis is unclear and biologic-based classifiers are not routinely applied at diagnosis. Circulating tumor DNA (ctDNA) is a highly tumor-specific biomarker that is prognostic in aggressive B-cell lymphomas, but its ability to serially monitor FL remains undefined. We applied a next-generation sequencing assay to identify tumor clonotypes for serial monitoring of peripheral blood in pts with untreated FL as part of an ongoing prospective clinical trial [NCT03190928]. Methods: Pts with grade I-II or 3A FL are eligible if evaluable disease on CT or FDG-PET, age ≥18, ECOG ≤2, no evidence of HT, and no prior systemic therapy. Pts undergo W&W until they meet uniform protocol-defined treatment criteria and remain on study until second-line therapy. Baseline testing includes labs, peripheral blood flow cytometry, BM biopsy/aspirate, CT and FDG-PET scans, and research biopsy. Pt have clinic visits every 4m for 2y, every 6m in years 3-5, then annually. CT scans are every 8m for 2y, then annually. FDG-PET scans are at baseline, at 2y, and any time of suspected progression. Peripheral blood samples including Streck tubes (plasma) and PBMCs are drawn at each clinic visit and stored. For ctDNA analysis, tumor DNA was amplified from FFPE using locus-specific primer sets for the Ig heavy-chain and light-chain loci along with BCL1/BCL2 translocations (Adaptive Biotechnologies). Amplified products were sequenced and tumor clonotypes were identified in plasma and PBMCs. Serial tracking of ctDNA was done in plasma and blinded to clinical outcomes. Results: 77 pts enrolled between July 2017 and July 2021. Median age was 57 (range 24-83) including 14 (18%) low-risk, 29 (38%) intermediate-risk, and 34 (44%) high-risk by FLIPI. Fourteen (18%) pts had stage I-II disease. Forty-three (56%) pts had monoclonal B-cells on peripheral blood flow cytometry. Twenty-nine (38%) pts progressed requiring frontline therapy including 7 (9%) pts with HT. Twenty-five (32%) pts were monitored ≥2y with no progression including 10 (13%) pts with evidence of at least some spontaneous regression by CT. Twenty (26%) pts were on study for Conclusions: ctDNA quantified from plasma in FL mirrors TMTV. Serial monitoring of ctDNA in patients without therapy demonstrated various patterns of fluctuation, including some patients in which ctDNA became undetectable coincident with spontaneous clinical regressions. ctDNA thus provides a non-invasive platform to monitor the natural history of FL, enabling future studies of tumor immune surveillance in this disease. Figure 1 Figure 1. Disclosures Jacob: Adaptive Biotechnologies: Current Employment, Current equity holder in publicly-traded company. Bagaev: BostonGene Corp.: Current Employment, Current holder of stock options in a privately-held company, Patents & Royalties: BostonGene. Meerson: BostonGene: Current Employment, Current holder of stock options in a privately-held company, Patents & Royalties: BostonGene. Postovalova: BostonGene Corp.: Current Employment, Current holder of stock options in a privately-held company, Patents & Royalties: BostonGene. Kudryashova: BostonGene: Current Employment, Current holder of stock options in a privately-held company, Patents & Royalties: BostonGene. Kotlov: BostonGene Corp: Current Employment, Current holder of stock options in a privately-held company, Patents & Royalties. Fowler: BostonGene: Current Employment, Current holder of stock options in a privately-held company.

While Epstein-Barr virus (EBV) and the Kaposi sarcoma herpesvirus (KSHV)/human herpesvirus (HHV) 8 have shown a definite association with lymphoproliferative disease, a role for the HHV-6 has been less clear. Similar to other herpesviruses, HHV-6 predominantly remains latent following initial infection, but can be reactivated during stress or immune suppression, and is the cause of roseola in young children. Existing as two distinct species, HHV-6B is more common, infecting ~90% of adults. HHV-6B, a T-lymphotropic virus, enters cells via CD134, a TNF receptor superfamily member, expressed on both naïve and CD4 +CD25 + T cells, leading to CD4 + lymphocyte depletion and impaired T cell activation. HHV-6 has been variably detected in classic Hodgkin (CHL) and T-cell lymphomas (TCL) by immunohistochemistry (IHC) and PCR with more recent data suggesting infection may be confined to tumor-associated lymphocytes. The specificity of these IHC antibodies is not well documented. The question remains whether HHV-6 in the tumor microenvironment of advanced disease is a consequence of immune dysfunction, or may play a more direct role in tumor initiation and progression by altering the tumor microenvironment. To address these questions, we evaluated HHV-6B viral gene expression patterns in lymphoma patient samples by RNA sequencing techniques. Following IRB approval, CHL, TCL, B-cell, and post-transplant lymphoproliferative disease (PTLD) cases were screened for potential HHV-6-association by IHC with an antibody against HHV-6 gp60/110 envelope glycoprotein (Millipore Sigma, MAB8537). Positive cases with available frozen tissue and adequate RNA (5) or sorted T-cell subsets from Hodgkin lymphoma (11) underwent bulk RNA-seq (rRNA depletion (Illumina), 50M reads/sample). Viral transcripts were identified by performing the Burrows-Wheeler Alignment by reference host alignment (to filter host and bad quality reads) followed by viral reference host alignment. Previous TCL databases with available RNAseq data were similarly evaluated. IHC analysis revealed 5/25 CHL, 34/52 TCL, 5/13 PTLD, 4/81 diffuse large B-cell lymphoma (DLBCL) and 2/28 follicular lymphoma (FL) with rare gp60/110-positive cells. This included 11 CHL cases with sorted T-cell subsets, of which one showed membranous and Golgi gp60/110 staining in background T-cells (25-year-old female, nodular sclerosis subtype, EBV-negative). Of these 11 CHL cases, RNAseq of T-cell subsets revealed a pattern of HHV-6B transcripts in only this case. Frozen tumor blocks were available from 5 additional cases with positive gp60/110 staining (2 CHL, 1 DLBCL, 1 FL and 1 PTLD), but RNAseq analysis did not identify any HHV-6B transcripts. Notably, these cases had dim cytoplasmic but not Golgi gp60/110 staining. RNA sequencing data derived from two independent TCL cohorts were analyzed for HHV-6B transcripts. Although no HHV-6B transcripts were detected via RNAseq in 20 angioimmunoblastic T-cell lymphoma samples from one TCL cohort, many had EBV-gene expression. HHV-6B transcripts were detected in two cases of anaplastic large cell lymphoma (ALCL) in a second TCL cohort (2/79 cases). High expression of the U67, U68, U79 and U90 genes was found, revealing overlap of the HHV-6B transcript expression between ALCL and CHL samples (Fig 1). Additionally, detection of two genes that could be driving tumor growth (U51, which encodes a G-protein receptor and U24, which inhibits proper T cell activation, reducing secretion of cytokines at infection site) demonstrates a specific viral gene expression pattern within the intratumor T-cell population. The potential presence of HHV-6B infection in the lymphoma microenvironment is controversial. To our knowledge, this is the first report conclusively demonstrating HHV-6B expression in CHL using RNAseq. Notably, the viral gene expression pattern seen in CHL overlaps with that found in two cases of ALCL, highlighting viral proteins of potential particular significance. These data may aid in development of a more reliable means of HHV-6B detection. For example, the immediate early gene U90, a transcriptional activator that may induce expression of both viral and cellular genes that affect the tumor microenvironment, was consistently expressed and may be a reliable marker of HHV-6B infection. Funding: HHV-6 Foundation Figure 1 Figure 1. Disclosures Tychinin: BostonGene Inc.: Current Employment, Current holder of stock options in a privately-held company, Patents & Royalties. Karelin: BostonGene Inc.: Current Employment, Current holder of stock options in a privately-held company, Patents & Royalties. Cherdintsev: BostonGene Inc.: Current Employment, Current holder of stock options in a privately-held company, Patents & Royalties. Kudryashova: BostonGene: Current Employment, Current holder of stock options in a privately-held company, Patents & Royalties: BostonGene. Egorov: BostonGene: Current Employment, Current holder of stock options in a privately-held company, Patents & Royalties. Degryse: BostonGene Inc.: Current Employment, Current holder of stock options in a privately-held company. Kotlov: BostonGene Corp: Current Employment, Current holder of stock options in a privately-held company, Patents & Royalties. Bagaev: BostonGene Corp.: Current Employment, Current holder of stock options in a privately-held company, Patents & Royalties: BostonGene. Roth: Merck: Consultancy; Janssen: Consultancy. Roshal: Celgene: Other: Provision of services; Physicians' Education Resource: Other: Provision of services; Auron Therapeutics: Other: Ownership / Equity interests; Provision of services. Rabadan: Genotwin: Other: Raul Rabadan is founder of Genotwin; AimedBio: Membership on an entity's Board of Directors or advisory committees. Elemento: Owkin: Consultancy, Other: Current equity holder; Freenome: Consultancy, Other: Current equity holder in a privately-held company; Volastra Therapeutics: Consultancy, Other: Current equity holder, Research Funding; One Three Biotech: Consultancy, Other: Current equity holder; Janssen: Research Funding; Eli Lilly: Research Funding; Champions Oncology: Consultancy; AstraZeneca: Research Funding; Johnson and Johnson: Research Funding.

Background: Diffuse large B-cell lymphoma (DLBCL) subtypes have differential response to BTK inhibitors (BTKi). Ibrutinib with R-CHOP improves survival in DLBCL subsets, but toxicity is limiting. Precise characterization of BTKi-responsive tumors enhances pt selection. Acalabrutinib (acala) is a BTKi with activity in DLBCL, but the molecular correlates of acala response are unknown. Circulating tumor DNA (ctDNA) is a prognostic biomarker in DLBCL including early changes during chemotherapy. PhasED-Seq is a novel ctDNA method that lowers the error profile of mutation detection by requiring the concordant detection of two separate mutations on an individual cell-free DNA molecule (Kurtz et al. Nat Biotechnol 2021). We employed a response-adapted study of acala for up to 14d prior to frontline therapy for aggressive B-cell lymphoma to determine the molecular profile of BTKi-responsive tumors. We report preliminary results including dynamic changes in ctDNA from this ongoing trial [NCT04002947]. Methods: Pts with untreated aggressive B-cell lymphoma and any HIV status are eligible if age ≥18, ≥stage 2, PS ≤2, and adequate organ function. Pts with PMBL, unmeasurable lesions, or active CNS disease are excluded. Screening includes labs, CT and FDG-PET, BM, and CSF with flow cytometry. Pts first receive acala 100mg twice daily x 14d. Pts with Results: 34 pts enrolled between August 2019 and July 2021 and completed the acala window. Median age was 64 (range 28-85) including 13 (38%) < 60y, 14 (41%) 61-69, and 7 (21%) ≥70y. Three (9%) pts had HIV and 17 (50%) were high-risk by IPI. The median diagnosis to treatment was 22.5d (4-53). IHC subtypes by Hans included 17 (50%) non-GCB, 16 (47%) GCB, and 1 (3%) T-cell/histiocyte-rich large B-cell lymphoma (TRLBCL). Four (12%) pts were high-grade B-cell lymphoma with MYC and BCL2 and/or BCL6 (HGBL-DH). Fifteen (44%) pts responded to acala during the window, while 19 (56%) pts had no response (Figure 1A). Acala responses were seen across DLBCL subtypes including 7 (47%) pts with non-GCB, 7 (47%) pts with GCB, and 1 (7%) pt with TRLBCL. Twenty pts had RNA sequencing to confirm cell-of-origin including 10 responders which included 7 (70%) GCB, 2 (20%) ABC, and 1 (10%) Unclassified. Notably, 13 (86%) BTKi-responsive tumors were CD10 negative and only 2 (18%) CD10+ tumors were BTKi-responsive. ctDNA dynamics strongly correlated with CT response as the log-fold change in ctDNA (hGE/mL) at the end of the window correlated with change on CT (r=0.75, p=0.0013). Remarkably, ctDNA dynamics after only 7d also correlated with change on CT (r=0.82, p=0.00006)(Figure 1B-C). Interestingly, one pt had improved symptoms and a 20-fold drop in ctDNA, but no corresponding CT changes suggesting that ctDNA changes may precede CT changes in some cases. Twenty-nine (85%) pts completed all planned cycles of therapy while 5 pts stopped chemotherapy early due to myelosuppression (n=3), CHF (n=1), and MI (n=1). Toxicity across 156 cycles was mostly hematologic. G3/G4 neutropenia occurred in 50% and 38% of cycles and febrile neutropenia in 10% of cycles. G3/G4 thrombocytopenia occurred in 22% and 12% of cycles. No increase in infections, atrial fibrillation, or bleeding were observed in pts treated with acala. All 27 pts who completed therapy achieved a CR. Two pts (1 acala responder) have relapsed from CR and 1 pt died of an MI. After a median follow-up of 9.2m the estimated 1-year PFS was 84.9% (95% CI: 58-95). Conclusions: Acalabrutinib prior to frontline therapy has activity in GCB, non-GCB, and HGBL-DH: confirmed by gene expression profiling. CD10+ GCB tumors are mostly acala-resistant. Toxicity is mainly hematologic and manageable across age groups including pts with HIV. ctDNA correlates with CT change and may predict response to targeted agents as early as 7 days. Updated clinical results within genetic subtypes will be presented at the meeting. Figure 1 Figure 1. Disclosures Chabon: Foresight Diagnostics: Current Employment, Current holder of individual stocks in a privately-held company, Current holder of stock options in a privately-held company. Lurain: CTI Biopharma: Research Funding; EMD-Serrono: Research Funding; Merck: Research Funding; BMS-Celgene: Research Funding; Janssen: Research Funding. Bagaev: BostonGene Corp.: Current Employment, Current holder of stock options in a privately-held company, Patents & Royalties: BostonGene. Postovalova: BostonGene Corp.: Current Employment, Current holder of stock options in a privately-held company, Patents & Royalties: BostonGene. Meerson: BostonGene: Current Employment, Current holder of stock options in a privately-held company, Patents & Royalties: BostonGene. Kudryashova: BostonGene: Current Employment, Current holder of stock options in a privately-held company, Patents & Royalties: BostonGene. Kotlov: BostonGene Corp: Current Employment, Current holder of stock options in a privately-held company, Patents & Royalties. Fowler: BostonGene: Current Employment, Current holder of stock options in a privately-held company. Kurtz: Roche: Consultancy; Foresight Diagnostics: Consultancy, Current holder of stock options in a privately-held company; Genentech: Consultancy. Alizadeh: CAPP Medical: Current holder of individual stocks in a privately-held company, Current holder of stock options in a privately-held company; Forty Seven: Current holder of individual stocks in a privately-held company, Current holder of stock options in a privately-held company; Foresight Diagnostics: Consultancy, Current holder of individual stocks in a privately-held company, Current holder of stock options in a privately-held company; Roche: Consultancy, Honoraria; Janssen Oncology: Honoraria; Celgene: Consultancy, Research Funding; Gilead: Consultancy; Bristol Myers Squibb: Research Funding; Cibermed: Consultancy, Current holder of individual stocks in a privately-held company, Current holder of stock options in a privately-held company.

When detecting explosive traces by optical remote methods based on recording the vapors, it is essential to know the applicability limits of these techniques. The amount of a vaporized explosive depends on the vaporization kinetics, which, in turn, relies on the physicochemical properties of the explosive, film weight and thickness, ambient air temperature, and film temperature. In active detection techniques, the explosive film temperature on the surface of objects may be higher than the air temperature due to special heating devices. In this case, the questions on how the quantity of matter available for detection is about to change and how much energy has to be consumed to heat up the film can be answered by mathematical modeling. The mathematic model is based on the Hertz–Knudsen–Langmuir equation that describes the vaporization rate of matter with mass transfer between the surface and surrounding air, which is taken into account. In an elaboration of the mathematical model, we introduced previously the temperature difference between the film and ambient air, which is now taken into consideration. The basic parameters influencing the film vaporization rate, and their variation range, were identified. The kinetic parameters of vaporization of thin films of some explosives with a quantity of matter typical of a fingerprint were estimated. The weight of matter in air during vaporization of explosive thin films in a wide range of parameters under study was calculated. Conclusions were made of the applicability limits of the developed standoff detection methods for trace explosives.

Gastric Cancer (GC) is the second leading cause of cancer-related mortality (9.7% of the total) and most patients with advanced disease will die within one year of diagnosis. GC is histologically classified into intestinal, diffuse and the mixed types, and into four molecular subtypes based on genetic profiling (i.e. microsatellite instable (MSI), EBV positive, chromosomal instable, and genomically stable). Although the molecular annotation is meaningful, the tumor immune microenvironment (TIME) has largely not been evaluated. Using an integrated collection of 11 public cohorts (n=2,270), we identified 5 unique GC TIME phenotypes by unsupervised clustering of gene expression signatures of RNA that describe the composition and biology of TIME and properties of malignant cells: two stromal-enriched subtypes - 1) Mesenchymal-wnt activated, high stroma activation and WNT signaling and 2) Fibrotic, only high stroma activation; two immune enriched subtypes 3) Inflamed non-fibrotic, high immune infiltration and low stromal compartment and 4) B-cell inflamed, high B cell activation; and 5) Immune Depleted, lowest immune and the highest malignant cell properties. The clusters were robust, being identified across all 11 datasets and across all stages of disease. Intestinal, diffuse, and mixed histologies were identified in each cluster. These clusters will be orthogonally validated using immunohistochemistry. The current molecular subtypes were represented in each of our TIME clusters, with some enrichment. Specifically, we found enrichment of MSI molecular subtype, characterized by hypermutation and microsatellite instability, in “Inflamed, non-fibrotic” cluster whereas the MSS/EMT subtype, associated with poor overall survival, was enriched in “Mesenchymal, wnt activated” cluster. We also saw specific enrichment of EBV positive tumors, known to have good prognosis, in both immune-enriched clusters. These results show a high concordance with the current TCGA/ACRG molecular subtypes of GC. These TIME clusters are prognostic in GC. The “Inflamed, non-fibrotic” cluster demonstrated a better overall and relapse free survival whereas stromal enriched clusters exhibited the worst (p We have defined and characterized the TIME for GC. The GC microenvironment is both prognostic for patient outcome and predictive of response to cytotoxic therapy. Citation Format: Prashant V. Thakkar, Olga Kudryashova, Daria Melikhova, Naira Samarina, Sandrine Degryse, Alexander Bagaev, Felix Frenkel, Svetlana Podsvirova, Dmitry Tychinin, Sandipto Sarkar, Rhonda K. Yantiss, Nathan Fowler, Manish A. Shah. Tumor immune microenvironment based molecular functional clustering reveals a prognostic signature that predicts overall survival in patients with gastric cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 3182.

Al based materials including Al-nanodiamonds and Al-Al2O3 processed by explosive compaction are considered in this paper. Improvements of composite properties were obtained by the introduction of diamond and ceramic nanoparticles (10 wt% of each) into the mixture of aluminium nanopowder. Various schemes and conditions, including a theoretical calculation of the explosion process, were suggested and discussed. Physical and mechanical properties of composites reinforced with nanoparticles are determined and discussed with special consideration paid to process conditions. The results show an increase in the hardness, compressive strength and elastic modulus of the composites after explosive compaction with the introduction of nanoparticles into the powder mixture. Improvement of mechanical properties of composites compared to pure aluminium is determined by dispersion strengthening, small material grain size and internal compressive stresses caused by dynamic shock-wave compaction processing.

Vibration treatment of solidifying metals results in improvement in the ingot structure. There is a need to study this process not only because of the practical potential of vibration treatment but also due to the lack of understanding the process. An important practical challenge is to find optimal conditions for liquid metal processing. In this paper, the authors consider a solidification process in the particular case of a cylindrical chill mold with vibration as a solution of the Stefan problem. An integral value of mechanical stresses in the melt during solidification is considered as an efficiency criterion of vibration treatment. A dependence of this value on the vibration frequency and amplitude is obtained through solving the Stefan problem numerically. The solution allows one to find the optimal vibration frequency and amplitude. We verified the numerical solution with experimental data obtained upon vibration treatment of aluminum melt under different conditions. The experimentally found optimal conditions for metal processing were similar to those proposed in theory, i.e., a vibration frequency of about 60 Hz and an amplitude of about 0.5 mm.

Here we look into the spread of aerosols indoors that may potentially carry viruses. Many viruses, including the novel SARS-CoV-2, are known to spread via airborne and air-dust pathways. From the literature data and our research on the propagation of fine aerosols, we simulate herein the carryover of viral aerosols in indoor air. We demonstrate that a lot of fine droplets released from an infected person’s coughing, sneezing, or talking propagate very fast and for large distances indoors, as well as bend around obstacles, lift up and down over staircases, and so on. This study suggests equations to evaluate the concentration of those droplets, depending on time and distance from the source of infection. Estimates are given for the safe distance to the source of infection, and available methods for neutralizing viral aerosols indoors are considered.

It is known that the use of external effects, such as acoustic fields (from ultrasonic to low-frequency range), help in breaking down agglomerates, improving particle wettability, providing uniform particle distribution in the melt volume, and reducing the grain size. The fragmentation of growing crystals, de-agglomeration of particles and their mixing in liquid metal under the influence of vibration (with frequencies of 10–100 Hz) are considered in this paper. The major advantage of such a technique in comparison with high-frequency methods (sonic, ultrasonic) is the capability of processing large melt volumes proportional to the wavelength. The mechanisms of the breaking down of particle agglomerates and the mixing of particles under conditions of cavitation and turbulence during the vibration treatment of the melt are considered. Expressions linking the threshold intensity and frequency with the amplitude necessary to activate mechanisms of turbulence and cavitation were obtained. The results of vibration treatment experiments for an aluminum alloy containing diamond nanoparticles are given. This treatment makes it possible to significantly reduce the grain size and to improve the casting homogeneity and thus improve the mechanical properties of the alloy.

Here we discuss the problem of atomizing submicron aerosols by special design atomizers enabling the cavitation regime. The formation of submicron aerosol was studied using an impulse atomizer model powered by a high-energy material and an atomizer model with a special spray nozzle generating countercurrent flows. For these atomizers, the role played by cavitation in producing submicron liquid aerosols is demonstrated herein. A mathematical model is also suggested to describe the aerosol cloud genesis. The cavitation development critical pressure, outflow velocity, and the resulting droplet sizes were evaluated. The aerosol particle size and concentration were experimentally measured by optical methods. The measured disperse parameters of aerosols during the origination and propagation of the aerosol cloud resulted from the cavitation-assisted atomization of liquids are reported: the intrinsic particle diameter of water aerosol is 10…30 μm depending on the features of the atomizer designs and their operating regimes.

A theoretical estimation of extent of influence of physical and chemical properties and concentration of the weighed particles on evolution of parameters of a liquid-droplets aerosol is carried out in the work. A series of experiments on a research of parameters of aerosol media with a various structure and concentration characteristics of a dispersive phase under ex-ternal (ultrasonic and electrostatic) fields were made.

Advanced air quality control requires real-time monitoring of particulate matter size and concentration, which can only be done using optical instruments. However, such techniques need regular calibration with reference samples. In this study, we suggest that puffball fungus (Lycoperdon pyriforme) spores can be utilized as a reference standard having a monodisperse size distribution. We compare the Lycoperdon pyriforme spores with the other commonly used reference samples, such as Al2O3 powder and polystyrene latex (PSL) microspheres. Here we demonstrate that the puffball spores do not coagulate and, thus, maintain the same particle size in the aerosol state for at least 15 minutes, which is enough for instrument calibration. Moreover, the puffball mushrooms can be stored for several years and no agglomeration of the spores occurs. They are also much cheaper than other calibration samples and no additional devices are needed for aerosol generation since the fungal fruiting body acts as an atomizer itself. The aforementioned features make the fungal spores a highly promising substance for calibration and validation of particle size analyzers, which outperforms the existing, artificially produced particles for aerosol sampling. Furthermore, the L. pyriforme spores are convenient for basic research and development of new optical measurement techniques, taking into account their uniform particle size and absent coagulation in the aerosol.

This article is about new constitutional order in Russia emphasizing fundamental areas as the role of judiciary in the Russian Constitutional System from 1993 Constitution, the one which replaced the 1978 Russian Soviet Federative Socialist Republic Constitution based on communist ideology. Therefore, it is intended to examine the principle of separation of powers, constitutionally recognized human rights and liberties, relationship between international bodies’ case law of human rights and the Russian Constitutional Court, among others issues. El presente artículo se detiene a analizar el papel de la judicatura en el sistema constitucional ruso a partir de la Constitución de 1993, aquella que sustituyó a la Constitución de 1978 de la República Socialista Federativa de la Unión Soviética que estaba basada en la ideología comunista. Así, con la finalidad de dar cuenta del nuevo orden constitucional, enfatiza en importantes aristas como el principio de separación de poderes, los derechos humanos y libertades reconocidos constitucionalmente, la interrelación entre la jurisprudencia de los órganos internacionales de derechos humanos y la Corte Constitucional de Rusia, entre otros.

This work considers the deagglomeration and wettability of particles by metal melt and proposes a mechanism of particle agglomerate dispersion by ultrasonic cavitation. The main dependences connecting the processing time and intensity with the physical and chemical properties of particles and the melt as well as acoustic parameters are obtained. For the first time found that melt during ultrasonic treatment, inclusive the particles agglomerates proportional to melt viscosity and the size of the agglomerates. It has been established that time ultrasonic treatment melt containing the particles agglomerates is proportional to melt viscosity and the size of the agglomerates. The required time for successful melt infiltration in the agglomerates, wettability and their introduction into the melt takes ten minutes. The suggested equation allows estimating the intensity of ultrasonic radiation, required to destroy the agglomerates of particles in the melt. It was found that intensity of the ultrasound must be inversely proportional to the radius of the agglomerates. The theoretical results are confirmed by comparing with experimental dates.

In the manuscript process of crystallization of metals at ultrasonic influence is considered. The known models of crystallization under the influence of ultrasound are investigated. By means of calculations influence of parameters of ultrasonic influence and characteristics of crystals of metal on the received structure of alloy is established. The corresponding mathematical model of ultrasonic crushing of particles and their agglomerates in metal fusion is offered. It is determined the threshold mode of ultrasonic influence on fusion at which almost monodisperse distribution of particles by the sizes is reached.

After the onset of a long-term low dose rate radiation exposure (55-60 y later) of the Techa riverside residents within a range of individual red bone marrow (RBM) doses from 0.01 to 1.79 Gy, it was established that there was an obvious association between the type of reaction manifested by peripheral blood lymphocytes to small dose irradiation in vitro (adaptive potential) and the RBM cell composition (during the period of the major exposure), as well as the peripheral blood cell composition (at a late time period coincident with the studies of induced radioresistance). The nature of these dependencies observed in chronically exposed individuals differs from that revealed in the controls. Based on the results of the study, it can be suggested that the capacity for adaptive response shown by peripheral blood lymphocytes donated by exposed persons at late time after exposure can be regarded as a biological marker of the functional state of the hemopoietic stem cell pool.

Polydisperse gas flows with condensed particles suspended therein are widely spread in nature and play an important role in many branches of the modern engineering and technology. There is a necessity for estimating parameters of disperse media in the manufacturing processes, scientific research, and atmosphere sounding. In industry, the result of this estimation can be employed as the quality measure of a product. When designing and distributing multi-phase systems to study the processes of controlling their parameters, it is also needed to determine values of the quantities characterizing such systems.