Active Bone Marrow Research Articles

An Atypical Presentation of Alcoholic Hepatitis

Significant alcohol consumption leads to a variety of alcoholic liver diseases ranging from alcoholic fatty liver disease to alcoholic cirrhosis. Alcoholic hepatitis carries a high risk of mortality; therefore, it is important to identify patients early to guide treatment and management. We discuss an atypical presentation of alcoholic hepatitis, with initial concern for lymphoproliferative disease. Our patient was a 28—year—old Cantonese—American man presenting with one month of abdominal pain, fevers, night sweats, and weight loss. His social history was not impressive for chronic alcohol use;he was a social drinker who drank 1 beer and 3 whisky shots every 4 months. Physical exam revealed tachycardia, abdominal tenderness, hepatosplenomegaly, and diffuse jaundice with no stigmata of chronic liver disease. Labs revealed leukocytosis with white blood cell count of 22x103/uL, conjugated hyperbilirubinemia (total bilirubin:4.4mg/dL), elevated alkaline phosphatase(183 U/L), and an INR of 1.43. An infectious process was ruled out when cultures and CXR were unrevealing. An abdominal ultrasound revealed hepatomegaly and ascites, but no ductal dilatation,which was confirmed on MRCP. Given his age, presence of B symptoms, and lack of significant alcohol history, there was concern for underlying malignancy. A CT abdomen showed abdominal lymphadenopathy and hepatosplenomegaly with a subsequent PET—CT scan showing diffuse lymphadenopathy and increased bone marrow activity. Hematology performed a bone marrow biopsy that revealed no signs of a neoplastic process. Patient's bilirubin continued to rise to 20mg/dL so a transjugular liver biopsy was obtained. Measurements revealed portal hypertension with HVPG of 10 and histology demonstrated steatosis, mild portal fibrosis,and Mallory body formation with neutrophilic infiltrates, suggestive of alcoholic hepatitis. The patient's Maddrey Discriminant Function score was elevated at 34 and he was started on prednisolone. At day 7 his Lille score was 0.04, indicating response to steroids. He completed a 28—day course of prednisolone and had complete resolution of fevers, ascites and biochemical abnormalities, while remaining abstinent. This is a case of alcoholic hepatitis presenting atypically with B symptoms, lack of strong drinking history, and diffuse lymphadenopathy, all suggestive of lymphoma. This case highlights the importance of pursuing a liver biopsy early on to confirm a diagnosis of alcoholic hepatitis.

Initial Report of a Phase I Study of LY2510924, Idarubicin, and Cytarabine in Relapsed/Refractory Acute Myeloid Leukemia.

Background: The CXCR4/SDF-1α axis plays a vital role in the retention of stem cells within the bone marrow and downstream activation of cell survival signaling pathways. LY2510924, a second generation CXCR4, showed significant anti-leukemia activity in a murine AML model.Methods: We conducted a phase I study to determine the safety and toxicity of LY2510924, idarubicin and cytarabine (IA) combination therapy in relapsed/refractory (R/R) AML. Eligible patients were 18–70 years of age receiving up to salvage 3 therapy. A peripheral blood absolute blast count of < 20,000/μL was required for inclusion. LY2510924 was administered daily for 7 days followed by IA from day 8. Two dose escalation levels (10 and 20 mg) were evaluated, with a plan to enroll up to 12 patients in the phase I portion.Results: The median age of the enrolled patients (n = 11) was 55 years (range, 19–70). Median number of prior therapies was 1 (1–3). Six and five patients were treated at dose-levels “0” (10 mg) and “1” (20 mg), respectively. Only one patient experiencing a dose limiting toxicity (grade 3 rash and myelosuppression). Three and one complete responses were observed at dose-levels “0” and “1,” respectively; the overall response rate (ORR) was 36% (4 of 11 patients). A ≥ 50% decrease in CXCR4 mean fluorescence intensity was observed in 4 of 9 patients by flow cytometry, indicating incomplete suppression of CXCR4-receptor occupancy.Conclusions: The combination of LY2510924 with IA is safe in R/R AML. Dose-escalation to a 30 mg LY2510924 dose is planned to achieve complete blockade of CXCR4 receptor occupancy, followed by expansion phase at the recommended phase 2 dose-level.

Hematologic Perturbations among Pesticide Applicators Recently Exposed to Permethrin in the Agricultural Health Study

In the Agricultural Health Study (AHS), a large prospective cohort of pesticide applicators, exposure to the pyrethroid insecticide permethrin is associated with an increased risk of multiple myeloma (MM), a hematologic malignancy with an unfavorable prognosis. To gain insight into the biological mechanisms underlying this association, we assessed potential hematotoxic effects of permethrin, in a longitudinal study of 35 permethrin-exposed male pesticide applicators in the AHS. Serial blood samples were collected from each participant in the off-season, the day after permethrin use, and approximately 3 weeks after use. At each time point, we performed a complete blood count with white blood cell differential and measured lymphocyte subsets. We used linear mixed models to assess the relationship between permethrin exposure and cell counts, controlling for season, age, BMI, recent cold/infection (&amp;#8804;7 days), and use of other pesticides (&amp;#8804;30 days). We observed a statistically significant increase in the geometric mean of immature granulocytes (31%, 95% CI&amp;#61;6%, 63%) on the day after permethrin use relative to off-season levels, but not 3 weeks after use. Statistically significant (P&amp;lt;0.05) alterations in several red blood cell parameters were also observed on the day after permethrin use, including decreased red blood cell count, hemoglobin, and hematocrit and increased mean corpuscular volume and red blood cell distribution width-SD. Red blood cell count, hemoglobin, and hematocrit remained significantly decreased 3 weeks after permethrin use. Increased immature granulocytes are indicative of an inflammatory response and enhanced bone marrow activity, and sustained decreases in red blood cell parameters may be a result of increased oxidative stress. Our results suggest inflammatory and hematologic effects of permethrin exposure, and identify targeted cell populations for future mechanistic studies to elucidate the relationship between permethrin use and MM risk.

Correlation of bone marrow cellularity and metabolic activity in healthy volunteers with simultaneous PET/MR imaging.

To evaluate the correlation between bone marrow cellularity (BMC) and metabolic activity in healthy subjects and to see whether yellow marrow is indeed metabolically quiescent. Because metabolic activity can be assumed to reflect vascularity, we assessed the relationship between regional metabolic activity and geographic frequency of metastases as noted in the literature. Two hundred and twenty locations (ten in each side of the pelvis and proximal femur) were evaluated in 11 consecutive healthy volunteers with simultaneous PET/MR. BMC was calculated through precise water-fat fraction quantification with a 6-echo gradient echo. We analyzed correlations between cellularity and SUVr, age, and R2*. We also looked at the relation between our results and the reported prevalence of metastases. There was moderate but statistically significant correlation between BMC and metabolic activity (r = 0.636, p < 0.0001). Interestingly, the iliac and sacrum had higher metabolic activity relative to cellularity, whereas the femoral neck and lesser trochanter showed lower SUVr than other regions with the similar cellularity. The relatively lower metabolic status of the femoral neck conflicted with its reported high frequency of metastasis. Excluding regions with almost no remaining red marrow, cellularity showed inverse relationship with age (r = 0.476, p < 0.0001) and direct relationship with R2* (r = 0.532, p < 0.0010). Metabolic activity of bone marrow was largely dependent on BMC while yellow marrow seems metabolically quiescent. The discrepancy between the assumed vascularity as determined by metabolic activity and reported sites of metastasis suggested that the process of bone metastasis may not depend entirely on vascularity.

A Review on Iron Oxide Nanoparticles and Their Biomedical Applications

The iron oxide nanoparticles have a great attraction in biomedical applications due to their non-toxic role in the biological systems. The iron oxide nanoparticles have both magnetic behaviour and semiconductor property which lead to multifunctional biomedical applications. The iron oxide nanoparticles used in biomedical fields such as antibacterial, antifungal and anticancer were reviewed. The uses of hematite (α-Fe2O3), maghemite (γ-Fe2O3) and magnetite (Fe3O4) nanoparticles, for an inhibition time in biological activities, are listed in this work. Also, this review explains the use of iron oxide nanoparticles in the biomedical fields with particular attention to the application of hematite and superparamagnetic iron oxide nanoparticles. In this review, analysis reveals that the role of iron oxide in biological activity is good due to its biocompatibility, biodegradability, ease of synthesis and different magnetic behaviours. The change of properties of iron oxide nanoparticles such as particle size, morphology, surface, agglomeration and electronic properties has specific impact in biomedical application. The review mainly focused in and discussed about antibacterial, anticancer, bone marrow and cell labelling activities. From this review work, the iron oxide nanoparticle may be specialised in particular bacterial and cancer treatments. Also discussed are the iron oxide nanoparticle-specific biomedical applications like human placenta, insulin and retinal locus treatments.

Identification of an Early Unipotent Neutrophil Progenitor with Pro-tumoral Activity in Mouse and Human Bone Marrow

Neutrophils are short-lived cells that play important roles in both health and disease. Neutrophils and monocytes originate from the granulocyte monocyte progenitor (GMP) in bone marrow; however, unipotent neutrophil progenitors are not well defined. Here, we use cytometry by time of flight (CyTOF) and single-cell RNA sequencing (scRNA-seq) methodologies to identify a committed unipotent early-stage neutrophil progenitor (NeP) in adult mouse bone marrow. Importantly, we found a similar unipotent NeP (hNeP) in human bone marrow. Both NeP and hNeP generate only neutrophils. NeP and hNeP both significantly increase tumor growth when transferred into murine cancer models, including a humanized mouse model. hNeP are present in the blood of treatment-naive melanoma patients but not of healthy subjects. hNeP can be readily identified by flow cytometry and could be used as a biomarker for early cancer discovery. Understanding the biology of hNeP should allow the development of new therapeutic targets for neutrophil-related diseases, including cancer.

2007 - Interferon Gamma Increases Hematopoietic Stem Cell Homing to an Activated Bone Marrow Niche

2007 - Interferon Gamma Increases Hematopoietic Stem Cell Homing to an Activated Bone Marrow Niche

Leukaemia and myeloid malignancy among people exposed to low doses (

Substantial evidence links exposure to moderate or high doses of ionising radiation, particularly in childhood, with increased risk of leukaemia. The association of leukaemia with exposure to low-dose (<100 mSv) radiation is less certain, although this is the dose range most relevant to the general population. We aimed to estimate the risk of leukaemia associated with low-dose radiation exposure in childhood (age <21 years). In this analysis of historical cohort studies, we pooled eligible cohorts reported up to June 30, 2014. We evaluated leukaemia and myeloid malignancy outcomes in these cohorts with the relevant International Classification of Diseases and International Classification of Diseases for Oncology definitions. The cohorts included had not been treated for malignant disease, had reported at least five cases of the relevant haematopoietic neoplasms, and estimated individual active bone marrow (ABM) doses. We restricted analysis to individuals who were younger than 21 years at first irradiation who had mean cumulative ABM doses of less than 100 mSv. Dose-response models were fitted by use of Poisson regression. The data were received in fully anonymised form by the statistical analyst. We identified nine eligible cohorts from Canada, France, Japan, Sweden, the UK, and the USA, including 262 573 people who had been exposed to less than 100 mSv enrolled between June 4, 1915, and Dec 31, 2004. Mean follow-up was 19·63 years (SD 17·75) and mean cumulative ABM dose was 19·6 mSv (SD 22·7). 154 myeloid malignancies were identified (which included 79 acute myeloid leukaemias, eight myelodysplastic syndromes, and 36 chronic myeloid leukaemias, in addition to other unspecified myeloid malignancies) and 40 acute lymphoblastic leukaemias, with 221 leukaemias (including otherwise unclassified leukaemias but excluding chronic lymphocytic leukaemia) identified overall. The fitted relative risks at 100 mSv were 3·09 (95% CI 1·41-5·92; ptrend=0·008) for acute myeloid leukaemia and myelodysplastic syndromes combined, 2·56 (1·09-5·06; ptrend=0·033) for acute myeloid leukaemia, and 5·66 (1·35-19·71; ptrend=0·023) for acute lymphoblastic leukaemia. There was no clear dose-response for chronic myeloid leukaemia, which had a relative risk at 100 mSv of 0·36 (0·00-2·36; ptrend=0·394). There were few indications of between-cohort heterogeneity or departure from linearity. For acute myeloid leukaemia and myelodysplastic syndromes combined and for acute lymphoblastic leukaemia, the dose-responses remained significant for doses of less than 50 mSv. Excess absolute risks at 100 mSv were in the range of 0·1-0·4 cases or deaths per 10 000 person-years. The risks of acute myeloid leukaemia and acute lymphoblastic leukaemia were significantly increased after cumulative doses of ionising radiation of less than 100 mSv in childhood or adolescence, with an excess risk also apparent for cumulative radiation doses of less than 50 mSv for some endpoints. These findings support an increased risk of leukaemia associated with low-dose exposure to radiation and imply that the current system of radiological protection is prudent and not overly protective. National Cancer Institute Intramural Research Program, National Cancer Institute, and US National Institutes for Health.

Abstract CT145: A Cancer Research UK phase I trial of anti-GD2 chimeric antigen receptor (CAR) transduced T-cells (1RG-CART) in patients with relapsed or refractory neuroblastoma

Abstract Background: Treatment of high risk neuroblastoma remains challenging; current multimodal treatment regimens achieve long term survival in &amp;lt;50% of patients and are associated with significant morbidity. Ganglioside GD2 is abundantly expressed on almost all neuroblastomas whilst expression on normal tissue is highly limited, providing a suitable CAR target. Here, we report the preliminary results of a Phase I clinical study of GD2-CART for refractory/relapsed neuroblastoma (NCT02761915). Trial design: The therapeutic (1RG-CART) is autologous T-cells transduced with a gamma-retroviral vector encoding both an anti-GD2 CAR and the RQR8 suicide gene. The CAR comprises a humanized anti-GD2 single chain variable fragment derived from the K666 antibody and CD28/CD3ζ signalling domains. Both lymphodepletion and CART dose were escalated as follows: dose level (DL) 1 without lymphodepletion, DL2 with 1.2 g/m2 cyclophosphamide and DL3 and beyond 1.2 g/m2 cyclophosphamide and 125 mg/m2 fludarabine followed by administration of a single intravenous dose of 1x107/m2 (DL1-3) or 1x108/m2 (DL4) 1RG-CART. Primary objectives are to assess safety and tolerability. Results: To date, 12 patients have been enrolled. All had relapsed/refractory neuroblastoma with measurable disease in bone (n=11), bone marrow (n=7) and/or soft tissue sites (n=9). Cell products were successfully manufactured for all patients. Median transduction efficiency was 34.5% (range 16-54%). Nine patients have been treated on DL1 (n=4), DL2 (n=1), DL3 (n=1) and DL4 (n=3) respectively. No dose limiting toxicity (DLT) was seen. For patients treated on DL1-3 (1x107/m2), 1RG-CART could not be detected in peripheral blood, and no clinical responses were seen. In contrast, expansion of 1RG-CART cells as detected by flow cytometry and qPCR was seen in the 3 patients treated on DL4 (1x108/m2). In two DL4 patients, 1RG-CART expansion was still limited and transient (marking levels &amp;lt;10,000 copies/μg DNA). These patients had disease progression as measured at Day +28. In one DL4 patient however, 1RG-CART marking levels of &amp;gt;40,000 copies/μg DNA were achieved. This patient developed Grade 2 cytokine release syndrome (Day +5) and biochemical evidence of tumour lysis (Day +21). Disease reassessment on Day +28 showed response in many sites of bone/marrow disease as measured by mIBG scintigraphy, and near complete tumour clearance in bone marrow which at baseline was heavily infiltrated with neuroblastoma. Disease progression occurred on Day +45 at which time 1RG-CART were no longer detectable. In the absence of DLT this prompted us to continue with DL5 (1x109/m2). Conclusions: These preliminary results are the first to demonstrate on-target activity in bone and bone marrow of GD2-CART in this childhood solid tumour. Further 1RG-CART dose escalation is warranted, and under way. Citation Format: Karin Straathof, Barry Flutter, Rebecca Wallace, Simon Thomas, Gordon Cheung, Angela Collura, Talia Gileadi, Jack Barton, Gary Wright, Sarah Inglott, David Edwards, Claire Barton, Karen Dyer, Nigel Westwood, Thalia Loka, Sarita Depani, Karen Howe, Giuseppe Barone, Martin Pule, John Anderson. A Cancer Research UK phase I trial of anti-GD2 chimeric antigen receptor (CAR) transduced T-cells (1RG-CART) in patients with relapsed or refractory neuroblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr CT145.

Abstract 37: Pulmonary pre-metastatic modifications induced by breast cancer

Abstract Breast cancer can be highly curable at early stages, while the survival rate plummets with the occurrence of metastasis. Thus, studies of molecular mechanisms that allow cancer cells to leave the primary tumor and ultimately establish colonies at distant sites are of crucial importance. First proposed by Stephen Paget in 1889, the “seed and soil” hypothesis suggests that the secondary organ microenvironment undergoes constant modification induced by the primary tumor through circulating chemokines. This leads to the migration of activated bone marrow derived cells to secondary organs, increased matrix metalloproteases (MMPs), and extracellular matrix (ECM) modifications in the secondary organ, collectively termed the pre-metastatic niche, which provides desirable docking sites for cancer cells. Our previous studies have shown persistent changes in the collagenous stroma and proteoglycans in pre-metastatic lungs of mice growing metastatic breast tumors, whose extent directly correlated with the malignancy of the primary tumor [1]. Elevated proteolytic MMP activity was also detected in these pre-metastatic lungs [2]. Based on these findings, we have continued our pre-metastatic niche studies by building an appropriate cell culture model, which consists of lung fibroblasts that were exposed to cell culture media conditioned by differentially malignant human breast epithelial cells. Cell lines used for fibroblast conditioning were nonmalignant MCF-12A breast epithelial cells (control), non-metastatic MCF-7 breast cancer cells, and highly metastatic, triple-negative MDA-MB-231 breast cancer cells. We focused our studies on ECM- and MMP-related changes, which we analyzed using qRT-PCR and immunoblotting. qRT-PCR analysis revealed that metastatic MDA-MB-231 cells induced an increase in the ECM protein Col14A1 as well as MMP-2 and MMP-14 in lung fibroblast as compared to induction by MCF-12A control. Immunoblotting analyses of lung fibroblasts showed that breast cancer cells induced increases in MMP expression in lung fibroblasts. Even higher MMP levels in fibroblasts were induced by metastatic MDA-MB-231 cells as compared to non-metastatic MCF-7 cells. Breast cancer cells also induced decreased expression levels of secreted MMPs in lung fibroblasts, most likely due to an enhanced release of these MMPs. This decrease was most significantly induced by the metastatic cell line. In conclusion, we have observed elevated levels in ECM molecules and MMPs in lung fibroblasts upon exposure of conditioned media from breast cancer cells. Our findings point towards an important role of lung fibroblasts in the evolution of the pre-metastatic niche in modifying the ECM, and their ability to directly respond to factors released from primary breast cancer cells with which they are not in direct contact. [1] Paidi SK et al. Cancer Res. 2017; 77:247-56. [2] Rizwan et al. Npj Breast Cancer 2015; 1: 15017. Citation Format: Ruoqing Cai, Caitlin M. Tressler, Menglin Cheng, Kanchan Sonkar, Santosh K. Paidi, Ishan Barman, Kristine Glunde. Pulmonary pre-metastatic modifications induced by breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 37.

Kelch-like protein 14 promotes B-1a but suppresses B-1b cell development.

B-1 cells are innate-like B-cell population and produce natural antibodies that contribute to the first line of host defense. There are two subsets of B-1 cells: B-1a and B-1b. B-1a cells are the main producer of poly-reactive and autoreactive natural IgM antibodies, whereas B-1b cells can respond specifically to T-cell-independent antigens. Despite the functional significance of B-1a and B-1b cells, little information is available about what regulates the development of these two subsets. We found that Kelch-like protein 14 (KLHL14) was expressed at high levels in B cells but only at low levels in a few non-lymphoid tissues. Although mice lacking KLHL14 died right after birth, the heterozygotes developed normally with no gross abnormalities by appearance. B-cell development in the bone marrow and maturation and activation in the spleen were not affected in the heterozygous mice. However, the number of peritoneal B-1a cells was significantly reduced while B-1b cells were increased in Klhl14 heterozygous mice compared with wild-type (WT) mice. Consistently, Rag1-/- mice reconstituted with Klhl14-/- fetal liver cells had a more severe reduction of B-1a and an increase of B-1b cells in the peritoneal cavity. KLHL14 did not affect the turnover or apoptosis of B-1a and B-1b cells in vivo. Moreover, Klhl14-/- fetal liver contained a similar proportion and absolute numbers of the B-1 progenitor cells as did WT fetal liver. These results suggest that KLHL14 promotes B-1a development in mice.

Dose to Pelvic Bone Marrow Defined with FDG-PET Predicts for Hematologic Nadirs in Anal Cancer Patients Treated with Concurrent Chemo-radiation

Purpose: To investigate whether irradiated volume of pelvic active bone marrow (ACTBM) may predict decreased blood cells nadirs in anal cancer patients undergoing concurrent chemo-radiation.Methods: Forty-four patients were analyzed and pelvic active bone marrow (ACTBM) was characterized employing 18FDG-PET. Dosimetric parameters on dose–volume histograms were correlated to nadirs with generalized linear modeling.Results: ACTBM mean dose was significantly correlated to white blood cell (β = −1.338; 95%CI: −2.455/−0.221; p = 0.020), absolute neutrophil count (β = −1.651; 95%CI: −3.284/−0.183; p = 0.048), and platelets (β = −0.031; 95%CI: −0.057/−0.004; p = 0.024) nadirs. Other dosimetric parameters were found to be correlated (ACTBM-V10,-V20,-V30and-V40).Conclusions: 18FDG-PET is able to define active bone marrow and may predict for decreased blood cells count nadirs.

Evaluation of the rivastigmine role against botulinum toxin-A-induced osteoporosis in albino rats: A biochemical, histological, and immunohistochemical study.

The present study aimed to evaluate the role of rivastigmine against the effect of a single unilateral botulinum toxin-A (BTX-A) injection on the bone and bone marrow of adult albino rats 4 weeks after injection. Twenty-four Wistar albino rats were divided into four equal groups: group I, which received distilled water; group II, which received rivastigmine (0.3 mg/kg daily, intraperitoneally for 4 weeks); group III, which received BTX-A (4 IU in 0.2 mL physiological saline) single dose, intramuscularly; and group IV, which received BTX-A + rivastigmine. The results revealed that BTX-A induced a significant decrease in the calcium level with a significant increase in the phosphorus, alkaline phosphatase, C-reactive protein, and tumor necrosis factor α levels in serum. Furthermore, a significant increase in malondialdehyde with a significant decrease in reduced glutathione activities in both bone and bone marrow. Histologically, a distortion and thinning out of the compact bone and trabeculae of cancellous bone of the rat femur in the BTX-A group with an increase in adipocytes in adjacent bone marrow were detected. Immunohistochemically, Cluster of Differentiation 68 (CD68) showed a significant increase in both osteoclasts and bone marrow macrophage. Rivastigmine treatment could relieve the toxic effects induced by BTX-A. In conclusion, rivastigmine has a protective effect against the hazardous effects of BTX-A on bone and bone marrow.

Prolonged hematopoietic and myeloid cellular response in patients after an acute coronary syndrome measured with 18F-DPA-714 PET/CT

PurposeAn acute coronary syndrome (ACS) is characterized by a multi-level inflammatory response, comprising activation of bone marrow and spleen accompanied by augmented release of leukocytes into the circulation. The duration of this response after an ACS remains unclear. Here, we assessed the effect of an ACS on the multi-level inflammatory response in patients both acutely and after 3 months.MethodsWe performed 18F-DPA-714 PET/CT acutely and 3 months post-ACS in eight patients and eight matched healthy controls. DPA-714, a PET tracer binding the TSPO receptor and highly expressed in myeloid cells, was used to assess hematopoietic activity. We also characterized circulating monocytes and hematopoietic stem and progenitor cells (HSPCs) by flow cytometry in 20 patients acutely and 3 months post-ACS and in 19 healthy controls.ResultsIn the acute phase, patients displayed a 1.4-fold and 1.3-fold higher 18F-DPA-714 uptake in, respectively, bone marrow (p = 0.012) and spleen (p = 0.039) compared with healthy controls. This coincided with a 2.4-fold higher number of circulating HSPCs (p = 0.001). Three months post-ACS, 18F-DPA-714 uptake in bone marrow decreased significantly (p = 0.002), but no decrease was observed for 18F-DPA-714 uptake in the spleen (p = 0.67) nor for the number of circulating HSPCs (p = 0.75).Conclusions18F-DPA-714 PET/CT reveals an ACS- triggered hematopoietic organ activation as initiator of a prolonged cellular inflammatory response beyond 3 months, characterized by a higher number of circulating leukocytes and their precursors. This multi-level inflammatory response may provide an attractive target for novel treatment options aimed at reducing the high recurrence rate post-ACS.

A novel role for the adapter molecule NHERF1 in regulating mast cell-mediated anaphylaxis

Abstract Anaphylaxis is a life threatening allergic reaction that is classically elicited by crosslinking of antigen-specific IgE bound to the high affinity IgE receptor FcɛRI on mast cells. Proteases released from FcɛRI-activated mast cells result in the accumulation of the anaphylatoxin, C3a that can further amplify the anaphylactic response via interaction with C3a receptor (C3aR), a G-protein coupled receptor (GPCR). This autocrine loop resulting in C3aR activation as a consequence of FcɛRI stimulation initiates anaphylaxis in humans. C3aR possess a class I PDZ binding motif on its cytoplasmic tail. Adapter proteins such as the Na+/H+ exchanger regulatory factor 1 (NHERF1) have been implicated in regulating GPCR functions by binding to the class I PDZ motifs in these receptors. In our previous study, we had demonstrated that NHERF1 promotes C3a-induced degranulation and chemokine production in human mast cells. The aim of this study was to determine if NHERF1 regulates mouse mast cell response following FcɛRI stimulation. Our data suggests that NHERF1 enhances intracellular Ca2+ mobilization, activation of the MAP kinase pathway (ERK and P38) and production of cytokines (IL-6 and IL-13) following IgE/antigen-mediated activation in bone marrow derived mouse mast cells. In agreement with our in vitro data, NHERF1 promotes mast cell-mediated passive systemic anaphylaxis and passive cutaneous anaphylaxis induced by IgE and antigen in clinically relevant murine models. These results thus indicate that the NHERF1 plays a vital role in regulating mast cell-mediated anaphylaxis and that NHERF1 could be a potential novel target for anaphylaxis and other allergic diseases.

CCL4 enhances preosteoclast migration and its receptor CCR5 downregulation by RANKL promotes osteoclastogenesis

Chemokine CCL4 (MIP-1β) is released from osteoblast cells to restore the homeostasis of hematopoietic stem cells during the activation of bone marrow. In this study, we investigated the function of CCL4 and its receptor CCR5 during osteoclastogenesis. CCL4 promoted the migration and viability of preosteoclast cells. However, CCL4 had no direct effect on the receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclast differentiation in mouse preosteoclast cells. In addition, CCR5 expression was rapidly reduced by RANKL treatment, which was recovered by IFN-γ during osteoclastogenesis. CCR5 downregulation by RANKL was mediated by MEK and JNK in preosteoclast cells and promoted osteoclastogenesis. These results suggest that CCL4 can enhance the recruitment of preosteoclasts to bone in the early stage, and the reduction of CCR5 promotes osteoclastogenesis when RANKL is prevalent.

The indexes of osteogenic stem cells activity in patients with rheumatoid arthritis and frontal knee deformities

Objective: to study the indexes of stem cells activity of bone marrow which form knee joint (lateral and medial parts of distal femur and proximal tibia part) which were cut at arthroplasty surgery in patients with rheumatoid arthritis and frontal knee deformity.Methods: spongiest bone was taken at surgery (knee arthroplasty) from medial and lateral sides of distal femur and proximal tibia. Due to cloning method of fibroblast colony forming units we studied in 37 patients with rheumatoid arthritis of knee joint. We studied 168 samples of bone marrow, 204 cultures of stem cells were growth.Results: we observed significant deceleration of reparative process in bone tissue at rheumatoid arthritis. Significant difference of fibroblast colony forming units in 1 sm 3 volume in different parts of knee joint was observed. In the medial femur condylar this index was (0,2307 ± 0,16) · 104, it exceeds values obtained in other areas in 11–20 times. It was shown that stem cells of the lateral tibia condylar had worse conditions for regenerative potential in cases of valgus deformity compare with varus. There were 4 times less amount of fibroblast colony forming units and the efficacy of its cloning was 2.6 times lower.Conclusion: significant differences were revealed in the indexes of osteogenic activity of stem cells which form knee joint (lateral and medial parts of distal femur and proximal tibia part).

Serum-mediated Activation of Bone Marrow–derived Mesenchymal Stem Cells in Ischemic Stroke Patients

Stroke induces complex and dynamic, local and systemic changes including inflammatory reactions, immune responses, and repair and recovery processes. Mesenchymal stem cells (MSCs) have been shown to enhance neurological recovery after stroke. We hypothesized that serum factors play a critical role in the activation of bone marrow (BM) MSCs after stroke such as by increasing proliferation, paracrine effects, and rejuvenation. Human MSCs (hMSCs) were grown in fetal bovine serum (FBS), normal healthy control serum (NS), or stroke patient serum (SS). MSCs cultured in growth medium with 10% SS or NS exhibited higher proliferation indices than those cultured with FBS (P < 0.01). FBS-, NS-, and SS-hMSCs showed differences in the expression of trophic factors; vascular endothelial growth factor, glial cell–derived neurotrophic factor, and fibroblast growth factor were densely expressed in samples cultured with SS (P < 0.01). In addition, SS-MSCs revealed different cell cycle– or aging-associated messenger RNA expression in a later passage, and β-galactosidase staining showed the senescence of MSCs observed during culture expansion was lower in MSCs cultured with SS than those cultured with NS or FBS (P < 0.01). Several proteins related to the activity of receptors, growth factors, and cytokines were more prevalent in the serum of stroke patients than in that of normal subjects. Neurogenesis and angiogenesis were markedly increased in rats that had received SS-MSCs (P < 0.05), and these rats showed significant behavioral improvements (P < 0.01). Our results indicate that stroke induces a process of recovery via the activation of MSCs. Culture methods for MSCs using SS obtained during the acute phase of a stroke could constitute a novel MSC activation method that is feasible and efficient for the neurorestoration of stroke.

Assessment of PCXMC for patients with different body size in chest and abdominal x ray examinations: a Monte Carlo simulation study

A PC Program for x ray Monte Carlo (PCXMC) has been used to calculate organ doses in patient dosimetry and for the exposure assessment in epidemiological studies of radiogenic health related risks. This study compared the dosimetry from using the built-in stylized phantoms in the PCXMC to that of a newer hybrid phantom library with improved anatomical realism. We simulated chest and abdominal x ray projections for 146 unique body size computational phantoms, 77 males and 69 females, with different combinations of height (125–180 cm) and weight (20–140 kg) using the built-in stylized phantoms in the PCXMC version 2.0.1.4 and the hybrid phantom library using the Monte Carlo N-particle eXtended transport code 2.7 (MCNPX). Unfortunately, it was not possible to incorporate the hybrid phantom library into the PCXMC. We compared 14 organ doses, including dose to the active bone marrow, to evaluate differences between the built-in stylized phantoms in the PCXMC and the hybrid phantoms (Cristy and Eckerman 1987 Technical Report ORNL/TM-8381/V1, Oak Ridge National Laboratory, Eckerman and Ryman 1993 Technical Report 12 Oak Ridge, TN, Geyer et al 2014 Phys. Med. Biol. 59 5225–42). On average, organ doses calculated using the built-in stylized phantoms in the PCXMC were greater when compared to the hybrid phantoms. This is most prominent in AP abdominal exams by an average factor of 2.4-, 2.8-, and 2.8-fold for the 10-year-old, 15-year-old, and adult phantoms, respectively. For chest exams, organ doses are greater by an average factor of 1.1-, 1.4-, and 1.2-fold for the 10-year-old, 15-year-old, and adult phantoms, respectively. The PCXMX, due to its ease of use, is often selected to support dosimetry in epidemiological studies; however, it uses simplified models of the human anatomy that fail to account for variations in body morphometry for increasing weight. For epidemiological studies that use PCXMC dosimetry, associations between radiation-related disease risks and organ doses may be underestimated, and to a greater degree in pediatric, especially obese pediatric, compared to adult patients.

Chronic Alterations in Systemic Immune Function after Traumatic Brain Injury.

There is a compelling link between severe brain trauma and immunosuppression in patients with traumatic brain injury (TBI). Although acute changes in the systemic immune compartment have been linked to outcome severity, the long-term consequences of TBI on systemic immune function are unknown. Here, adult male C57Bl/6 mice underwent moderate-level controlled cortical impact (CCI) or sham surgery, and systemic immune function was evaluated at 1, 3, 7, 14, and 60 days post-injury. Bone marrow, blood, thymus, and spleen were examined by flow cytometry to assess changes in immune composition, reactive oxygen species (ROS) production, phagocytic activity, and cytokine production. Bone marrow derived macrophages (BMDMs) from sham and 60-day CCI mice were cultured for immune challenge studies using lipopolysaccharide (LPS) and interleukin-4 (IL-4) models. Acutely, TBI caused robust bone marrow activation and neutrophilia. Neutrophils and monocytes exhibited impairments in respiratory burst, cytokine production, and phagocytosis; in contrast, ROS levels and pro-inflammatory cytokine production were chronically elevated at 60 days post-injury. Cultures of BMDMs from chronic CCI mice demonstrated defects in LPS- and IL-4-induced polarization when compared with stimulated BMDMs from sham mice. TBI also caused thymic involution, inverted CD4:CD8 ratios, chronic T lymphopenia, greater memory conversion, increased T cell activation, impaired interferon γ induction, and chronically elevated Th1 cytokine and ROS production. Collectively, our in-depth phenotypic and functional analyses demonstrate that TBI induces widespread suppression of innate and adaptive immune responses after TBI. Moreover, at chronic time points, TBI mice exhibit hallmarks of accelerated immune aging, displaying chronic deficits in systemic immune function.