Cells In Circulation Research Articles

Clonally expanded CD38hi cytotoxic CD8 T cells define the T cell infiltrate in checkpoint inhibitor-associated arthritis.

Immune checkpoint inhibitor (ICI) therapies used to treat cancer, such as anti-PD-1 antibodies, can induce autoimmune conditions in some individuals. The T cell mechanisms mediating such iatrogenic autoimmunity and their overlap with spontaneous autoimmune diseases remain unclear. Here, we compared T cells from the joints of 20 patients with an inflammatory arthritis induced by ICI therapy (ICI-arthritis) with two archetypal autoimmune arthritides, rheumatoid arthritis (RA) and psoriatic arthritis (PsA). Single-cell transcriptomic and antigen receptor repertoire analyses highlighted clonal expansion of an activated effector CD8 T cell population in the joints and blood of patients with ICI-arthritis. These cells were identified as CD38hiCD127- CD8 T cells and were uniquely enriched in ICI-arthritis joints compared with RA and PsA and also displayed an elevated interferon signature. In vitro, type I interferon induced CD8 T cells to acquire the ICI-associated CD38hi phenotype and enhanced cytotoxic function. In a cohort of patients with advanced melanoma, ICI therapy markedly expanded circulating CD38hiCD127- T cells, which were frequently bound by the therapeutic anti-PD-1 drug. In patients with ICI-arthritis, drug-bound CD8 T cells in circulation showed marked clonal overlap with drug-bound CD8 T cells from synovial fluid. These results suggest that ICI therapy directly targets CD8 T cells in patients who develop ICI-arthritis and induces an autoimmune pathology that is distinct from prototypical spontaneous autoimmune arthritides.

The Weddell Gyre heat budget associated with the Warm Deep Water circulation derived from Argo floats

Abstract. The Weddell Gyre plays an important role in the global climate system by supplying heat to underneath the ice shelves and in the formation of deep and bottom water masses, which have been subject to widespread warming over past decades. In this study, we investigate the re-distribution of heat throughout the Weddell Gyre by diagnosing the terms of the heat conservation equation for a 1000 m thick layer of water encompassing the core of Warm Deep Water. The spatial distributions of the different advective and diffusive terms in terms of heat tendencies are estimated using gridded climatologies of temperature and velocity, obtained from Argo floats in the Weddell Gyre from 2002 to 2016. While the results are somewhat noisy on the grid scale and the representation of the effects of eddy mixing is highly uncertain due to the need to parameterise them by means of turbulent diffusion, the heat budget (i.e. the sum of all terms) closes (within the uncertainty range) when integrated over the open inflow region in the southern limb, whereas the interior circulation cell remains unbalanced. There is an overall balance in the southern limb between the mean horizontal advection and horizontal turbulent diffusion of heat, whereas the vertical terms contribute comparatively little to the heat budget throughout the Weddell Gyre. Heat convergence due to mean horizontal advection balances with divergence due to horizontal turbulent diffusion in the open southern limb of the Weddell Gyre. In contrast, heat divergence due to mean horizontal advection is much weaker than convergence due to horizontal turbulent diffusion in the interior circulation cell of the Weddell Gyre, due to large values in the latter along the northern boundary due to large meridional temperature gradients. Heat is advected into the Weddell Gyre along the southern limb, some of which is turbulently diffused northwards into the interior circulation cell, while some is likely turbulently diffused southwards towards the shelf seas. This suggests that horizontal turbulent diffusion plays a role in transporting heat both towards the gyre interior where upwelling occurs and towards the ice shelves. Horizontal turbulent diffusion is also a mechanism by which heat can be transported into the Weddell Gyre across the open northern boundary. Temporal deviations from the mean terms are not included due to study limitations. In order to appreciate the role of transient eddying processes, a continued effort to increase the spatial and temporal coverage of observations in the eastern Weddell Sea is required.

Acrolein produced by glioma cells under hypoxia inhibits neutrophil AKT activity and suppresses anti-tumoral activities

Acrolein, which is the most reactive aldehyde, is a byproduct of lipid peroxidation in a hypoxic environment. Acrolein has been shown to form acrolein-cysteine bonds, resulting in functional changes in proteins and immune effector cell suppression. Neutrophils are the most abundant immune effector cells in circulation in humans. In the tumor microenvironment, proinflammatory tumor-associated neutrophils (TANs), which are termed N1 neutrophils, exert antitumor effects via the secretion of cytokines, while anti-inflammatory neutrophils (N2 neutrophils) support tumor growth. Glioma is characterized by significant tissue hypoxia, immune cell infiltration, and a highly immunosuppressive microenvironment. In glioma, neutrophils exert antitumor effects early in tumor development but gradually shift to a tumor-supporting role as the tumor develops. However, the mechanism of this anti-to protumoral switch in TANs remains unclear. In this study, we found that the production of acrolein in glioma cells under hypoxic conditions inhibited neutrophil activation and induced an anti-inflammatory phenotype by directly reacting with Cys310 of AKT and inhibiting AKT activity. A higher percentage of cells expressing acrolein adducts in tumor tissue are associated with poorer prognosis in glioblastoma patients. Furthermore, high-grade glioma patients have increased serum acrolein levels and impaired neutrophil functions. These results suggest that acrolein suppresses neutrophil function and contributes to the switch in the neutrophil phenotype in glioma.

Correlation between admission red cell distribution width and GRACE risk score as an early mortality marker in patients with acute myocardial infarction

Background: Red blood cell distribution width (RDW) is a measurement of the size variation of red blood cells in circulation. The Global Registry of Acute Coronary Events (GRACE) score is the most commonly used risk-stratification scoring system to predict mortality in acute coronary syndrome for both ST- elevated myocardial infarction (STEMI), non-ST elevated myocardial infarction (NSTEMI). The aim of this study was to assess the easily determinable red blood cell distribution width (RDW) as an early mortality marker of acute myocardial infarction (MI) mortality and the correlation of RDW-CV on admission with GRACE risk score in patients with acute myocardial infarction. Methods: This hospital-based, prospective observational study included 211 patients (mean age 58.15 +/- 12.45 years, male 176) admitted with acute MI (STEMI/NSTEMI) from January 2022 to June 2022. The study population was divided into two groups basing on the 50th percentile of the admission RDW-CV value -13.7. The GRACE score was calculated on admission to predict the GRACE in hospital mortality score within 24 hours of admission. Results: The study found that the higher RDW-CV values were associated with adverse prognostic factors in acute myocardial infarction like increased age, lower systolic blood pressure, heart failure, cardiac arrest, higher cardiac biomarker levels and higher in-hospital mortality (p = 0.049, level of significance p< 0.05) and this correlated with a higher GRACE score (Pearson’s correlation coefficient R= 0.47, P<0.05). Conclusion: Thus, RDW can be used as a useful parameter for predicting in-hospital mortality in acute MI patients.

NUMERICAL study of MAGNETO convective Buongiorno nanofluid flow in a rectangular enclosure under oblique magnetic field with heat generation/absorption and complex wall conditions

A mathematical model is presented to analyze the magnetohydrodynamic (MHD) convective flow in a rectangular enclosure. Earlier studies on the Tiwai-Das volume fraction nanofluid model did not consider the Buongiorno nanofluid model. This is the focus of the present analysis which examines the laminar mixed convection magnetohydrodynamic flow of a nanofluid in a differentially heated rectangular enclosure with complex boundary conditions under an inclined magnetic field. Buongiorno's two-component nanofluid model is employed, which incorporates the effects of Brownian motion and thermophoretic diffusion of nanoparticles. Magnetic nanofluids have considerable potential for enhancing transport processes in energy systems such as hybrid fuel cells. The study is essential with heat generation/absorption effects. Additionally, the work is highlighted by the general case of an oblique (inclined) magnetic field. The conservation equations for mass, primary and secondary momentum, energy, and nanoparticle concentration with wall boundary conditions are dimensionless using appropriate scaling transformations. A finite-difference computational scheme known as the Harlow-Welch Marker and Cell (MAC) method is employed to solve the dimensionless nonlinear coupled boundary value problem. A mesh independence study is included. Graphical plots are presented for the impact of key control parameters on streamline contours, isotherm contours, iso-concentration (nanoparticle mass) contours, and local Nusselt number. With heat sink (absorption), the Nusselt number is enhanced in magnitude whereas it is suppressed with heat generation since there is a heat reduction transmitted to the boundary. The configurations of streamlines, isotherms, and iso-concentrations are mostly invariant to magnetic field direction changes. The obtained results show interesting behaviors of the flow and thermal fields, which mainly involve the effect of Brownian motion and thermophoresis parameters, as well as unsteady regimes, depending on specific values of the Schmidt number, Richardson number, and Prandtl numbers. Increasing the Schmidt number induces a contraction in the central cooler zone in the enclosure and also reduces the iso-concentration magnitudes in the central region across the enclosure. The core region of the enclosure heats up as the thermophoresis and Brownian motion parameters rise, pushing the previously cooler top and bottom wall zones further away from the center. There is also a decrease in iso-concentration magnitudes in particular at the upper and lower boundaries at higher values of Brownian motion and thermophoresis parameters. At decreasing buoyancy ratios, the left vortex cell first decelerates while the right vortex cell accelerates. However, when the buoyancy ratio increases, the left vortex cell streamlines magnitudes increase with a contraction in vortex size, while the right cell develops. A Very minor alteration is observed in the isotherm and iso-concentration contours with an increasing buoyancy ratio. When the Richardson number increases, the vortex cell structures shift from a strong circulation cell on the left to a weaker cell on the right, resulting in reverse distribution. With the rising Richardson number, significant cooling is also caused in the core zone, as well as a drop in iso-concentrations, with the original dual low-concentration upper and lower zones merging into a single center zone. The original symmetric left and right vortex cells are gradually twisted diagonally towards the right wall as the magnetic field increases, yet the stronger right cell and the weaker left cell are maintained.

Exogenous Antigen Upregulation Empowers Antibody Targeted Nanochemotherapy of Leukemia.

Acute myeloid leukemia (AML) is afflicted by a high-mortality rate and few treatment options. The lack of specific surface antigens severely hampers the development of targeted therapeutics and cell therapy. Here, it is shown that exogenous all-trans retinoic acid (ATRA) mediates selective and transient CD38 upregulation on leukemia cells by up to 20-fold, which enables high-efficiency targeted nanochemotherapy of leukemia with daratumumab antibody-directed polymersomal vincristine sulfate (DPV). Strikingly, treatment of two CD38-low expressing AML orthotopic models with ATRA and DPV portfolio strategies effectively eliminates circulating leukemia cells and leukemia invasion into bone marrow and organs, leading to exceptional survival benefits with 20-40% of mice becoming leukemia-free. The combination of exogenous CD38 upregulation and antibody-directed nanotherapeutics provides a unique and powerful targeted therapy for leukemia.

Gut dysbiosis-derived low-grade endotoxemia: A common basis for liver and cardiovascular disease.

Gut dysbiosis is characterized by bacteria overgrowth that ultimately leads to increased intestinal barrier permeability and translocation of bacteria or bacterial products such as lipopolysaccharide (LPS) in the portal and, eventually, systemic circulation. Intestinal epithelial cells and hepatocytes possess enzymatic armamentarium to counteract the LPS toxic effect, however, impaired degradation results in LPS accumulation in the hepatocytes and endothelial wall. Experimental and clinical studies documented that in patients with liver disease, such as nonalcoholic fatty acid liver disease (NAFLD), low-grade endotoxemia caused by LPS is implicated in liver inflammation and thrombosis via interaction with its Toll-like receptor 4 (TLR4) expressed by hepatocytes and platelets. Furthermore, studies in patients with severe atherosclerosis documented that LPS localizes in atherosclerotic plaque in close association with activated macrophages expressing TLR4 suggesting LPS's role in vascular inflammation, atherosclerotic progression, and thrombosis. Finally, LPS may directly interact with myocardial cells to induce electric and functional changes leading to atrial fibrillation or heart failure. This review will focus on experimental and clinical evidence suggesting that low-grade endotoxemia, as a mechanism, potentially accounts for vascular damage occurring at the level of the hepatic and systemic circulation and myocardial cells.

Increased tropical South Pacific western boundary current transport over the past century

The wind-driven meridional overturning circulation between the tropical and subtropical oceans is important for regulating decadal-scale temperature fluctuations in the Pacific Ocean and globally. An acceleration of the overturning circulation can act to reduce global surface temperature as ocean stores more heat. The equatorward low-latitude western boundary current represents a key component of the meridional circulation cell in the Pacific and a major source of water mass for the Equatorial Undercurrent, yet long-term observations of its transport are scarce. Here we demonstrate that the 15N/14N ratio recorded by Porites spp. corals in the western tropical South Pacific is sensitive to the exchanges of water masses driven by the western boundary transport. Using a 94-year coral record from the Solomon Sea, we report that the 15N/14N ratio declined as the global surface temperature rose. The record suggests that the South Pacific western boundary current has strengthened in the past century, and it may have contributed to the reported strengthening of the Equatorial Undercurrent. In addition, the 15N/14N record shows strong decadal variability, indicative of weaker equatorial Pacific upwelling and stronger western boundary transport when the eastern equatorial Pacific is in the warm stage of the Pacific Decadal Oscillation.

Estuarine nutrient pollution impact reduction assessment through euphotic zone avoidance/bypass considerations

The feasibility of reducing nutrient pollution impact by redirecting the effluent to depths below the euphotic zone was investigated for the deep estuarine Puget Sound region of the Salish Sea in the Pacific Northwest of America. The hypothesis tested was that the thickness of the outflow layer in deep estuaries may be greater than the euphotic zone depth, allowing a fraction of nutrients to be exported out passively through the layers immediately below. The euphotic zone depth in Puget Sound varies from 8 to 25 m while the depth of the outflow layer can reach up to ≈ 60 m. Outfall relocation strategies were tested on 99% of the anthropogenic nutrient loads currently delivered to Puget Sound. The impact was quantified using the previously established biophysical Salish Sea Model, using gross primary production and exposure to low dissolved oxygen (DO) levels as the metric (< 2 mg/L for hypoxia and < 5 mg/L for impairment). Eliminating nutrient pollution (above natural) from rivers and wastewater reduced hypoxia exposure by 8.1% and 11.2%, respectively. Relocating the outfalls to deeper waters resulted in improvements, but only in the sill-less sub-basins such as Whidbey, where hypoxia and DO impairment exposure decreased (7.9% and 6.8%, respectively). The presence of multiple sills and circulation cells in Puget Sound resulted in increased exposure and rendered nutrient bypass goals unfeasible as originally envisioned. However, an alternate nutrient export pathway was identified through bottom exchange flow out of Puget Sound via Whidbey Basin and Deception Pass. An unexpected reduction in the exchange outflow magnitude (≈ 4%) due additional (22%) freshwater discharged to the estuary bottom was also noted. The potential loss in circulation strength due to rerouting of natural surface freshwater through submerged deep-water outfalls is identified as a new unforeseen anthropogenic impact.

PD-1+ IFN-γ+ subset of CD8+ T cell in circulation predicts response to anti-PD-1 therapy in NSCLC.

Treatment with programmed cell death protein-1 (PD-1) antibodies has minimal response rates in patients with non-small cell lung cancer (NSCLC), and, actually, they are treated with chemotherapy combined with anti-PD-1 therapy clinically. Reliable markers based on circulating immune cell subsets to predict curative effect are still scarce. We included 30 patients with NSCLC treated with nivolumab or atezolizumab plus platinum drugs between 2021 and 2022. Whole blood was collected at baseline (before treatment with nivolumab or atezolizumab). The percentage of circulating PD-1+ Interferon-γ (IFN-γ+) subset of CD8+ T cell was determined by flow cytometry. The proportion of PD-1+ IFN-γ+ was calculated after gating on CD8+ T cells. Neutrophil/lymphocyte ratio (NLR), relative eosinophil count (%), and Lactate dehydrogenase (LDH) concentration at baseline of included patients were extracted from electronic medical records. The percentage of circulating PD-1+ IFN-γ+ subset of CD8+ T cell at baseline in responders was significantly higher than those in non-responders (P < 0.05). Relative eosinophil count (%) and LDH concentration in responders showed no significance between non-responders and responders. NLR in responders was significantly lower than those in non-responders (P < 0.05). Receiver operation characteristic (ROC) analysis found that the areas under the ROC curve for PD-1+ IFN-γ+ subset of CD8+ T cell and NLR were 0.7781 (95% CI, 0.5937-0.9526) and 0.7315 (95% CI, 0.5169-0.9461). Moreover, high percentage of PD-1+ IFN-γ+ subset in CD8+ T cells was relevant to long progression-free survival in patients with NSCLC treated with chemotherapy combined with anti-PD-1 therapy. The percentage of circulating PD-1+ IFN-γ+ subset of CD8+ T cell could be a potential marker at baseline to predict early response or progression in patients with NSCLC receiving chemotherapy combined with anti-PD-1 therapy.

Prophylactic simvastatin increased survival during endotoxemia and inhibited granulocyte trafficking in a cell-intrinsic manner

Cross sectional studies have shown that statin-users have improved odds of surviving severe sepsis. Meanwhile controlled clinical trials failed to demonstrate improved sepsis survival with acute statin administration following hospitalization. Here, a lethal murine peritoneal lipopolysaccharide (LPS) endotoxemia model was used to assess the efficacy of chronic versus acute simvastatin on survival. Mirroring clinical observations, chronic but not acute treatment with simvastatin significantly increased survival. At a pre-mortality time point in LPS-treated mice, chronic simvastatin suppressed granulocyte trafficking in to the lungs and peritoneum without otherwise suppressing emergency myelopoiesis, myeloid cells in circulation, or inflammatory cytokines. Chronic simvastatin treatment significantly downregulated inflammatory chemokine gene signature in the lungs of LPS-treated mice. Thus, it was unclear if simvastatin was inhibiting granulocyte chemotaxis in a cell intrinsic or extrinsic manner. Adoptive transfer of fluorescently labeled granulocytes from statin and vehicle treated mice into LPS-treated mice showed that simvastatin inhibited lung-granulocyte trafficking in a cell intrinsic manner. Congruent with this, chemotaxis experiments using in vitro macrophages and ex vivo granulocytes demonstrated that simvastatin inhibited chemotaxis in a cell-intrinsic manner. Collectively, chronic but not acute simvastatin treatment improved survival in murine endotoxemia, and this was associated with cell-intrinsic inhibition of granulocyte chemotaxis.

A Linear Theory of Local Thermal Circulations

The dynamics of local thermal circulations (LTCs) are examined by constructing a linear theory based on Boussinesq equations in the planetary boundary layer (PBL). Linear theory arranges LTCs into a sixth-order partial differential equation of temperature, which can be solved by using the Fourier transform and inverse Fourier transform. The analytic solution suggests that the horizontal distribution of the temperature anomaly is basically determined by surface heating, while the vertical distribution of the temperature anomaly is a combination of exponential decay and an Ekman spiral. For shallow PBL cases where the Ekman elevation is much smaller than the vertical scale of motion, the higher-order partial differential terms that represent the Ekman spiral structure can be ignored so that the equations reduce to a second-order partial differential equation. Compared with the numerical results, this so-called low-order approximation does not induce dramatic errors in the temperature distribution. However, to avoid distortion in the forced atmospheric circulation, the eddy viscosity in the motion equations should be replaced with the Rayleigh form, which is the common practice in LTCs. For deep PBL cases where the Ekman elevation is comparable to the vertical scale of motion, both the exponential decay and Ekman spiral structure play roles in the forced atmospheric circulation. The most significant influence is that there exist three additional compensating forced circulation cells that surround the direct forced circulation cell.

Abstract PR013: Unlocking the proteome of metastatic prostate cancer circulating tumor cells

Abstract In recent years, liquid biopsies have emerged to provide improved insights on the current disease state of a patient, but they generally rely on a small number of protein markers from immunofluorescent staining or cell-free DNA, which provides limited information especially around drug targets. As more targeted therapies and immunotherapies enter the market, selecting the optimal drug target could improve patient outcomes drastically. One way to achieve this goal is to generate multi-omic datasets from the cancer cells in circulation, known as circulating tumor cells (CTCs). In this study, we collaborated with Astrin Biosciences and used their patented isolation technology to enrich CTCs with high purity from the blood of 15 metastatic castration resistant prostate cancer (mCRPC) patients. The enriched CTCs and a patient matched control sample consisting primarily of white blood cells were divided such that RNA and protein could be interrogated from the same patient. Label free unbiased mass spectrometry-based proteomics was used to identify the CTC-enriched proteins and marks the first time a global proteomic assessment has been performed on CTCs from cancer patients. Initial analysis identified over 1,500 peptides corresponding to greater than 500 proteins from each patients’ CTCs; including increased epithelial cell markers in the CTC-enriched sample and increased WBC markers in the control sample. Hallmarks of androgen response and detection of the androgen receptor were also identified in the CTC-enriched proteome. Overall, this study was designed to show the feasibility of obtaining large scale proteomic data from a liquid biopsy and future studies will provide more insight into the clinical utility of such data with a focus on cancer signaling mechanisms and biomarkers. Importantly, as we analyze more patients, we plan to compare the proteomic data with future RNA signatures across mCRPC patients to eventually be able to correlate them with drug response with the overall goal to improve treatment selection and patient outcomes. Citation Format: Justin M. Drake, Zoi Sychev, Alec Horrmann, Kaylee Judith Smith, Catalina Galeano-Garces, Ali Arafa, Nicholas Heller, Mahdi Ahmadi, Jiarong Hong, Megan Ludwig, Justin Hwang, Emmanuel S. Antonarakis, Jayant Parthasarathy. Unlocking the proteome of metastatic prostate cancer circulating tumor cells [abstract]. In: Proceedings of the AACR Special Conference: Advances in Prostate Cancer Research; 2023 Mar 15-18; Denver, Colorado. Philadelphia (PA): AACR; Cancer Res 2023;83(11 Suppl):Abstract nr PR013.

Effect of circulating tumor cells distribution in treatment naive and treated patients with advance stage breast cancer on disease burden.

e12517 Background: Breast malignancies is a leading cause of cancer-related mortalities and show ascending incidence rate. Despite advancements in our understanding of the disease, its clinical outcome is often dismal. This largely remains owing to the characteristic wide window of relapse, spanning months to decades after primary treatment. Therefore, continuous monitoring of the disease is an offered choice to detect metastatic progression and recurrence. Circulating tumor cells (CTCs) have emerged as a powerful prognostic tool to predict the disease outcome in many epithelial cancers. CTCs are a real-time surrogate biomarker accounting for minimal residual disease (MRD) which is often missed in CT PET scanning. This leads to a progression of metastasis when the patient is often considered as ‘clinically disease free’. Here, we analyzed the presence of CTCs in treatment-naïve and chemo-recipient breast cancer patients. Methods: In this retrospective study on 417 breast cancer patients, CTCs were isolated from 1.5 ml of blood using the Drug Controller General of India (DCGI) approved OncoDiscover CTC test. This platform contains affinity-based magnetic nanoparticles to mediate EpCAM-based CTC isolation. CTCs were detected as CK18+, DAPI+ and CD45- cells based on automated digital imaging platform. Results: 42.6 % (n=178) of patients were clinically at progressive stage (stage II and III) and treatment naïve. On the other hand, 47.4 % of patients received treatment including surgery and chemotherapy. CTCs were not observed in 5.6 % (n=10) of the treatment naïve population while 32% (n=75) of patients who received therapy did not show CTCs. The mean CTC number in treatment naïve patients was 15, while the mean CTC count in patients receiving therapy was drastically reduced to 2. This implied that therapy effectively countered the tumor progression and reduced the shedding of tumor cells in circulation. The distribution of CTC in treatment naïve patients exhibited a bimodal trend centered at values of 10 and 50, suggesting two distinct populations of patients with respect to CTC count. CTC count did not show any correlation with the age in both population groups. Surprisingly CTC count in younger patients (20-50 years) was 50% higher compared to the older population (50-75 years). Conclusions: The presence of CTCs in treatment naïve, progressive breast cancer patients indicated the biologically aggravated disease. Although therapeutic intervention drastically reduced the CTC burden, their presence in a large population was suggestive of an MRD and the likelihood of recurrence after discontinuation of therapy. A distinct pattern of CTC occurrences in Tx naïve and Tx recipient patient suggested that CTCs can be an important clinical indicator to monitor the therapy response, progression and residual disease.

Correlation of micronuclei formation in circulating stromal cells with their PD-L1 expression and predicts for poorer clinical outcomes in metastatic breast cancer.

e14548 Background: Micronuclei (MN) are small, excised DNA fragments resulting from biological DNA repair mechanisms in response to internal chromosomal instability, such as those caused by genotoxic events (i.e. chemotherapy and radiation), and whose formation in tumor stromal cells in blood circulation is prognostic for worse clinical outcomes (Fenech 2020). Further, mutagenic events occurring during the formation of MN are associated with the development of cancer neoantigens, which are a major element in the programmed cell death ligand (PD-L1) pathway that influences response to immunotherapies. We hypothesized that MN can be detected in metastatic breast cancer (mBC) patients (pts) within circulating stromal cells (CStCs) that prognosticates for clinical outcomes and may associate with PD-L1 cell expression. Methods: We initiated a prospective pilot study using a compiled set of n=124 mBC samples, from multiple institutions, enrolled prior to starting new lines of treatment for progressive mBC. We enumerated MN formation found in CStCs from whole peripheral blood (7.5mL), filtered for CStCs, then stained for PD-L1 and DAPI. DAPI was used to identify MN, defined by small (&lt;3 µm) DAPI+ circular formations within the cytoplasm, separate from primary nuclei. In conjunction, all cells were stained with PD-L1 with expression normalized to background and evaluated against MN by T-test analysis. MN presence was compared to all available clinical variables, including induction of PD-L1 immunotherapy. Pts’ progression-free survival (PFS) and overall survival (OS) hazard ratios (HRs) were analyzed by censored univariate and multivariate analyses based on RECIST v1.1 over 24 months. Results: MN were identified in 56% (n=70/124) of CStCs and their presence was significantly prognostic for worse PFS (HR=2.4, CI= 1.53-3.76, p&lt;0.001) &amp; OS (HR= 1.89 CI=1.07-3.34, p=0.039) over 24 months. Overall, mPFS of pts with MN was 2.8 months and without MN was 12.1 months. The mOS of pts with MN was 12.7 months and without MN was 20.8 months. Additionally, MN positive CStCs had a significantly higher PD-L1 expression than MN negative CStCs (p&lt;0.001), with 67% of MN positive pts (n=46/66) having high PD-L1 expression while 30% of MN positive pts (n=20/66) having low PD-L1. Further, evaluation of PD-L1 in CStCs showed a linear relationship (R=0.9806) between MN number and PD-L1 in cells, with 0 MN average=0.016, 1 MN=0.200, 2 MN=0.318, and ≥3 MN=0.400. Conclusions: These initial findings suggest that MN are detected in pts with advanced mBC and may identify pts with faster progression and worse survival outcomes. Further, the number of MN in CStCs appears to correlate with higher PD-L1 expression within cells. Follow up clinical information and subtyping studies are ongoing to also evaluate of the effect of MN positive CStCs on PD-L1 immunotherapies in these pts.

Heart Failure Impairs Bone Marrow Hematopoietic Stem Cell Function and Responses to Injury.

Background Heart failure (HF) is a clinical syndrome associated with a progressive decline in myocardial function and low-grade systemic inflammation. Chronic inflammation can have lasting effects on the bone marrow (BM) stem cell pool by impacting cell renewal and lineage differentiation. However, how HF affects BM stem/progenitor cells remains largely unexplored. Methods and Results EGFP+ (Enchanced green fluorescent protein) mice were subjected to coronary artery ligation, and BM was collected 8 weeks after myocardial infarction. Transplantation of EGFP+ BM into wild-type mice revealed reduced reconstitution potential of BM from mice subjected to myocardial infarction versus BM from sham mice. To study the effects HF has on human BM function, 71 patients, HF (n=20) and controls (n=51), who were scheduled for elective cardiac surgery were consented and enrolled in this study. Patients with HF exhibited more circulating blood myeloid cells, and analysis of patient BM revealed significant differences in cell composition and colony formation potential. Human CD34+ cell reconstitution potential was also assessed using the NOD-SCID-IL2rγnull mouse xenotransplant model. NOD-SCID-IL2rγnull mice reconstituted with BM from patients with HF had significantly fewer engrafted human CD34+ cells as well as reduced lymphoid cell production. Analysis of tissue repair responses using permanent left anteriordescending coronary arteryligation demonstrated reduced survival of HF-BM reconstituted mice as well as significant differences in human (donor) and mouse (host) cellular responses after MI. Conclusions HF alters the BM composition, adversely affects cell reconstitution potential, and alters cellular responses to injury. Further studies are needed to determine whether restoring BM function can impact disease progression or improve cellular responses to injury.

Integrated analysis of single-cell and bulk RNA sequencing data reveals an immunostimulatory microenvironment in tumor thrombus of osteosarcoma

Tumor thrombus of bone sarcomas represents a unique reservoir for various types of cancer and immune cells, however, the investigation of tumor thrombus at a single-cell level is very limited. And it is still an open question to identify the thrombus-specific tumor microenvironment that is associated with the tumor-adaptive immune response. Here, by analyzing bulk tissue and single-cell level transcriptome from the paired thrombus and primary tumor samples of osteosarcoma (OS) patients, we define the immunostimulatory microenvironment in tumor thrombus of OS with a higher proportion of tumor-associated macrophages with M1-like states (TAM-M1) and TAM-M1 with high expression of CCL4. OS tumor thrombus is found to have upregulated IFN-γ and TGF-β signalings that are related to immune surveillance of circulating tumor cells in blood circulation. Further multiplexed immunofluorescence staining of the CD3/CD4/CD8A/CD68/CCL4 markers validates the immune-activated state in the tumor thrombus samples. Our study first reports the transcriptome differences at a single-cell level between tumor thrombus and primary tumor in sarcoma.

Effects of Intravascular Photobiomodulation on Insomnia, Muscle Soreness, and Biochemistry Profiles: An Eight-Year Retrospective Cohort.

Although cognitive-behavioral therapy is the first-line treatment for insomnia, pharmacotherapy is often prescribed to treat insomnia and related symptoms. In addition, muscle relaxants are commonly prescribed to alleviate muscle soreness when the pain is unbearable. However, pharmacotherapy can lead to numerous side effects. The non-drug strategy intravascular laser irradiation of blood (iPBM) has been advocated to improve pain, wound healing, blood circulation, and blood cell function to relieve insomnia and muscle soreness symptoms. Therefore, we assessed whether iPBM improves blood parameters and compared drug use before and after iPBM therapy. Consecutive patients who received iPBM therapy between January 2013 and August 2021 were reviewed. The associations between laboratory data, pharmacotherapies, and iPBM therapy were retrospectively analyzed. We compared patient characteristics, blood parameters, and drug use within the three months before the first treatment and the three months after the last treatment. We also compared the changes before and after treatment in patients who received ≥10 or 1-9 iPBM treatments. We assessed 183 eligible patients who received iPBM treatment. Of them, 18 patients reported insomnia disturbance, and 128 patients reported pain in any part of their body. After the treatment, HGB and HCT significantly increased after treatment in both the ≥10 and 1-9 iPBM treatment groups (HGB p < 0.001 and p = 0.046; HCT p < 0.001 and p = 0.029, respectively). Pharmacotherapy analysis revealed no significant differences in drug use before and after treatment, though drug use tended to decrease after iPBM. iPBM therapy is an efficient, beneficial, and feasible treatment that increases HGB and HCT. While the results of this study do not support the suggestion that iPBM reduces drug use, further larger studies using symptom scales are needed to confirm the changes in insomnia and muscle soreness after iPBM treatment.

Meteorological modeling of urban “heat islands” on the territory of Belarus under modern global warming

The article presents a quantitative estimation of intensity of urban “heat islands” in Belarus and their changes as a result of global warming. The potential opportunities for improving the urban microclimate by increasing the area of forest park zones in the city have been evaluated using mesoscale numerical simulation of meteorological conditions in cities. It is shown that in summer, Belarusian regional centers are warmer than suburban areas within 2–3 °С during the day and within 1.2–2.3 °С at night. Over the period that has passed since the beginning of this century, the regional cities have warmed by an average of 1.1–2.2 °C during the day and 1.3–1.9 °C at night. The maximum increase in daytime and nighttime temperatures in Belarusian cities has amounted within 3.1–6.0 and 2.4–4.4 °С, respectively. It is shown that replacing industrial clusters with forest park zones decreases the air temperature in the city within 2 °C and more than 2 times in evapotranspiration in the city of Homel. In the daytime, the northwest wind increases in the city and its suburb and this improves urban ventilation. At night, the afforestation of the city makes the surface layer of the atmosphere less stable, which is accompanied by the activation of local circulation cells, through which the temperature and specific humidity in the city and the suburb are smoothed.

Differential differentiation of B cell lymphopoiesis in lethal and non-lethal murine malaria models

ABSTRACT B cells in protection against malaria and need of experiencing many episodes in humans to achieve a state of immunity is largely unknown. The cellular basis of such defects in terms of B cell generation, maturation and trafficking was studied by taking Plasmodium chabaudi, a non-lethal and Plasmodium berghei, a lethal murine model. A flow cytometry (FCF) based evaluation was used to study alterations in generation and maintenance of B cells in patients with Plasmodium falciparum malaria as well as in murine malaria models. A significant accumulation of mature B cells in bone marrow and immature B cells in circulation was a feature observed only in lethal malaria. At peak parasitaemia, both the models induce a significant decrease in T2 (transitional) B cells with expansion of T1B cells. Studies in patients with acute Pf malaria showed a significant expansion of memory B cells and TB cells with a concomitant decrease in naive2 B cells as compared with healthy controls. This study clearly demonstrates that acute malarial infection induces major disturbances in B cell development in lymphoid organs and trafficking in periphery.