Bone Marrow Proliferation Research Articles

Cellular Proliferation of Equine Bone Marrow- and Adipose Tissue-Derived Mesenchymal Stem Cells Decline With Increasing Donor Age.

Background: Bone marrow (BM)- and adipose tissue (AT)-derived mesenchymal stem cells (MSCs) are used increasingly for autologous cell therapy in equine practice to treat musculoskeletal and other injuries. Current recommendations often call for 10–100 million MSCs per treatment, necessitating the expansion of primary cells in culture prior to therapeutic use. Of concern, human and rodent studies have shown a decline of both MSC recovery from sampled tissue and in vitro proliferative capacity with increasing donor age. This may be problematic for applications of autologous cell-based therapies in the important equine demographic of older patients.Objectives: To investigate the effect of donor age on the cellular proliferation of equine BM- and AT-MSCs.Study Design: In vitro study.Methods: BM- and AT-MSCs and dermal fibroblasts (biological control) were harvested from horses in five different age groups (n = 4, N = 60); newborn (0 days), yearling (15–17 months), adult (5–8 years), middle-aged (12–18 years), and geriatric (≥22 years). Proliferation of the cells was tested using an EdU incorporation assay and steady state mRNA levels measured for targeted proliferation, aging, and senescence biomarkers.Results: The cellular proliferation of equine BM- and AT-MSCs declined significantly in the geriatric cohort relative to the younger age groups. Proliferation levels in the two MSC types were equally affected by donor age. Analysis of steady state mRNA levels showed an up-regulation in tumor suppressors, apoptotic genes, and multiple growth factors in MSCs from old horses, and a down-regulation of some pro-cycling genes with a few differences between cell types.Main Limitations: Potential age-dependent differences in cell function parameters relevant to cell-therapy application were not investigated.Conclusions: The cellular proliferation of equine BM- and AT-MSCs declined at advanced donor ages. High levels of in vitro proliferation were observed in both MSC types from horses in the age groups below 18 years of age.

Multiple Myeloma with Suspected Non-Secretory Type

Multiple Myeloma (MM) is a hematological malignancy characterized by clonal plasma cell in bone marrow that produceabnormal globulin, which resulted in monoclonal gammopathy. Multiple Myeloma Non-Secretory (MMNS) is a very rareform of multiple myeloma with monoclonal plasmocytic proliferation in bone marrow supported by clinical manifestationand radiological findings. However, plasma cells fail to secrete immunoglobulin. A 44-year-old female came to SardjitoGeneral Hospital with main complaints of weakness and back pain. General weakness and pale palpebral conjunctiva were6 observed (+/+), liver and spleen were not palpable. Blood test results were as follows: Hb 3.0 g/dL, RBC 1.07 x 10 / μL, WBC3 3 562 x 10 /μL, PLT 114 x 10 /μL, A/G ratio 1.07, BUN 51.5 mg/dL, creatinine 4.62 mg/dL, and calcium 3.1 mmol/L. Skeletalsurvey suggested a multiple osteolytic. Protein electrophoresis revealed hypogammaglobulinemia with no M-spike. Therewere 66% of plasma cells in bone marrow. Patient was diagnosed by MMNS. Diagnosis MMNS can be established if clonalplasmacytes is accompanied with renal insufficiency and hypercalcemia. However, monoclonal gammopathy was not foundin serum protein electrophoresis. A case reported of 44-year-old female diagnosed as MMNS with 'punched out' multipleosteolytic, increased plasma cells in bone marrow without evidence of paraprotein in circulation proved by low A/G ratio andnegative M-spike.

Risk Factor for Erythropoietin Resistance in Hemodialysis Patient : Literature Review

Anaemia become the complication of chronic kidney disease (CKD) which was caused by decreasing of erythropoietin. Erythropoietin stimulating agent (ESA) therapy is one of therapy to overcome the problem, but until 34% of patients have lack of response to ESA treatment. Anaemia in CKD related to the worsen of the diseases, quality of life, and mortality of patients. Decreasing of the response to erythropoietin need to be evaluated to correct anaemic condition. This review is aim to explain the risk factor for erythropoietin resistance. The literature for this review was collected through PUBMED and google scholar. Erythropoietin is glycoprotein secreted 90% by interstitial cells of kidney and 10% by liver cells. The functions of erythropoietin are stimulate the proliferation and cells differentiation in bone marrow, and enhance erythropoiesis. Renal damage can inhibit the secretion of erythropoietin. In patients with ESA treatment, risk factors for resistance are iron deficiency, inadequate haemodialysis, inflammation, hyperparathyroid, nutrition disturbance, antibody mediated Pure Red Cell Aplastic (PRCA). The risk factor can be influenced by genetic variation. Conclusion of this review, there are several factor that influence the response of erythropoietin in hemodialisa patients. Hence, study related anaemia in CKD need further study to optimalize the treatment.

Adore: A Randomized, Open-Label, Phase 1/2 Open-Platform Study Evaluating Safety and Efficacy of Novel Ruxolitinib Combinations in Patients with Myelofibrosis

Adore: A Randomized, Open-Label, Phase 1/2 Open-Platform Study Evaluating Safety and Efficacy of Novel Ruxolitinib Combinations in Patients with Myelofibrosis

MiR-146a regulates osteogenic differentiation and proliferation of bone marrow stromal cells in traumatic femoral head necrosis.

The article "MiR-146a regulates osteogenic differentiation and proliferation of bone marrow stromal cells in traumatic femoral head necrosis, by Y. Kong, Z.-T. Chen, published in Eur Rev Med Pharmacol Sci 2019; 23 (2): 441-448-DOI: 10.26355/eurrev_201901_16853-PMID: 30720149" has been withdrawn from the authors stating that "some data cannot be repeated by our further research". The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/16853.

Screening of Feijoa (<i>Acca Sellowiana</i> (<i>O. Berg</i>) <i>Burret</i>) Fruit Effect on Proliferation and Apoptosis using Bone Marrow derived Stem Cells Model

Background and Objective: Curative properties of Feijoa sellowiana Bert. (Myrtaceae, Acca sellowiana), have been often claimed, while investigations into details of its bioactivities are still lacking. This study investigated the effect of acetonic extract of Feijoa (Acca sellowiana (O. Berg) Burret) fruit on proliferation and apoptosis of bone marrow derived stem cells. Methodology: Human bone marrow stem cells (hBMSCs) were characterized before experiments. MTT Assay was performed to determine the toxicity of Feijoa. The growth kinetics was also investigated until 7 days. Real time PCR assessed the expression of apoptosis genes. Results: In MTT assay, Feijoa at doses ≤ 5 ng/ml did not show any cytotoxic effect on hBMSCs and increased the cell proliferation until day 4th followed by a decrease until day 7th. Population doubling time (PDT) decreased until day 4th followed by an increase until day 7th. A significant increase in expression of Bax and decrease Bcl-2 expression were recorded on day 7th. Conclusion: The antioxidant, anti-inflammatory and modulatory activities of Feijoa can explain the increasing effect on cell proliferation till day 4th, but the apoptotic activity of Feijoa noted after four days is reported for the first time denoting to the short term antioxidant, anti-inflammatory and modulatory properties of Feijoa that should be considered for curative activity of this fruit.

Growth and proliferation of caprine bone marrow mesenchymal stem cells on different culture media

The growth and proliferation of mesenchymal stem cells are very sensitive in in vitro and a number of factors like media play a significant role in that context. In this study we assessed effect of different media on growth and proliferation of bone marrow derived mesenchymal stem cells (BMMSCs). The BMMSCs were isolated from caprine bone marrow and were subjected to magnetic activated cell sorting against CD90+, CD105+, CD271+and CD34− along with FC blocker. After characterisation, 2 × 104 cells were seeded in 12 well culture plates in four different media viz. MesenCult, MesenPRO, StemPro and complete DMEM (15 % FBS) to study their growth kinetic for 6 days from passage 0 (P0) to passage 3 (P3). The population doubling time (PDT) was derived from growth curve using logarithmic formula. The results showed that the BMMSCs growth and proliferation was highest in MesenCult media in P0 which varied significantly (p < 0.05) from rest of media and from P1 to P3, it was MesenPRO which yielded maximum cells (p < 0.05). The PDT was also in line with growth curve findings. In conclusion, the MesenPRO media had higher growth and proliferation rate from P1 to P3 although MesenCult had higher cell numbers in P0. In conclusion, the use of MesenPRO media could be a better option than conventional media when mesenchymal stem cells are used in clinical applications and other therapeutic purposes taking consideration to its higher growth and proliferation rate. And MesenCult would be a great option to harvest MSCs from P0.

Autotaxin inhibition reduces cardiac inflammation and mitigates adverse cardiac remodeling after myocardial infarction

ObjectiveAcute myocardial infarction (AMI) initiates pathological inflammation which aggravates tissue damage and causes heart failure. Lysophosphatidic acid (LPA), produced by autotaxin (ATX), promotes inflammation and the development of atherosclerosis. The role of ATX/LPA signaling nexus in cardiac inflammation and resulting adverse cardiac remodeling is poorly understood. Approach and resultsWe assessed autotaxin activity and LPA levels in relation to cardiac and systemic inflammation in AMI patients and C57BL/6 (WT) mice. Human and murine peripheral blood and cardiac tissue samples showed elevated levels of ATX activity, LPA, and inflammatory cells following AMI and there was strong correlation between LPA levels and circulating inflammatory cells. In a gain of function model, lipid phosphate phosphatase-3 (LPP3) specific inducible knock out (Mx1-Plpp3Δ) showed higher systemic and cardiac inflammation after AMI compared to littermate controls (Mx1-Plpp3fl/fl); and a corresponding increase in bone marrow progenitor cell count and proliferation. Moreover, in Mx1- Plpp3Δ mice, cardiac functional recovery was reduced with corresponding increases in adverse cardiac remodeling and scar size (as assessed by echocardiography and Masson's Trichrome staining). To examine the effect of ATX/LPA nexus inhibition, we treated WT mice with the specific pharmacological inhibitor, PF8380, twice a day for 7 days post AMI. Inhibition of the ATX/LPA signaling nexus resulted in significant reduction in post-AMI inflammatory response, leading to favorable cardiac functional recovery, reduced scar size and enhanced angiogenesis. ConclusionATX/LPA signaling nexus plays an important role in modulating inflammation after AMI and targeting this mechanism represents a novel therapeutic target for patients presenting with acute myocardial injury.

P6. A roughened 3D-printed surface enhances stem cell proliferation and osteoblast differentiation

BACKGROUND CONTEXT The advancement of manufacturing techniques, such as 3D-printing, has facilitated the design of complex and previously unattainable implant surface topographies. Research has shown surface properties, especially surface roughness, of an implant directly influence cellular response at the cell-material interface. Alkaline phosphatase (ALP), an early cellular marker of osteoblast differentiation, has been studied extensively to understand osteogenic cellular responses to a substrate. The identification of implant surfaces that promote a osteogenic environment may help identify those that are optimal for healing. PURPOSE The objective of this study was to investigate the effect of a roughened 3D-printed surface on the cellular proliferation and ALP expression of human bone-marrow derived mesenchymal stem cells (BM-MSCs). STUDY DESIGN/SETTING In vitro cell culture study. PATIENT SAMPLE N/A OUTCOME MEASURES Cellular proliferation was measured using a colorimetric, viability assay (alamarBlue™) to estimate the cell count on each substrate at days 1, 3 and 7. ALP activity was measured from cell lysate using a standard ALP assay on day 7. ALP activity was normalized to the total protein in each lysate sample measured using a bicinchoninic acid (BCA) assay also performed on day 7. METHODS Roughness (Ra) was characterized for each substrate in the study using a profilometer. BM-MSCs were seeded at a density of 10,000 cells/cm2 onto smooth polyetheretherkeytone (PEEK), smooth titanium-alloy (TAV), and a roughened, 3DP titanium-alloy (r3DP) substrates (n=6 per group) and cultured for 7 days. Cells were given 4 hours to attach, after which cell medium was changed and the substrates were moved to a separate well plate to remove any unattached cells. Assays were performed to quantify specific ALP cellular signaling, indicative of osteoblast differentiation. RESULTS The r3DP substrates had 15-30 times greater Ra than PEEK and TAV (15.2µm, 1.0µm, and 0.4µm, respectively). Cell count on day 7 for r3DP was significantly greater than PEEK (p 0.05). ALP activity was significantly greater in the r3DP group (8.8±1.1nmol/µg) compared to both PEEK (5.7±1.4nmol/µg) and TAV (4.4±0.9 nmol/µg) (p 0.05). CONCLUSIONS Optimal cellular activity and osteogenic cell signaling are important for osteoblast differentiation, a process needed to foster a cellular environment favorable for healing. Cells on a roughened 3D-printed TAV substrate exhibited significantly higher proliferation compared to PEEK and significantly higher ALP activity than both TAV and PEEK. Therefore, the r3DP substrate facilitated a cellular environment more favorable for osteoblast differentiation than TAV and PEEK substrates. Future research in an animal model is necessary to evaluate the downstream effects of an implant with a roughened 3D-printed TAV surface on the healing process. FDA DEVICE/DRUG STATUS SABLE (Globus Medical) (Approved for this indication)

Decoration of electrical conductive polyurethane-polyaniline/polyvinyl alcohol matrixes with mussel-inspired polydopamine for bone tissue engineering.

Electrospinning is a versatile technology for the fabrication of nanofibrous matrixes to regenerate defects. This study aims to develop a functionalized and electroconductive polymeric matrix to improve rat bone marrow mesenchymal stem cell adhesion, proliferation, and differentiation. Herein, the influence of the chemical composition of the substrate on homogeneous modification of the surface with mussel-inspired polydopamine (PDA) is focused. Accordingly, the deposition of PDA on the surface was proved by Fourier transform infrared spectroscopy. Morphologies of the scaffolds demonstrated homogeneous decoration of the polyvinyl alcohol (PVA)/polyurethane (PU)-polyaniline (PANI) matrixes with PDA, while a lower density of mussel-inspired polymer was observed in bare PU-PANI constructs. Although uniform and dense precipitation of PDA reduced conductivity of scaffolds 1.2 times compared with the samples with a low density of the PDA, 1.1 and 1.2 times enhancement in tensile strength and Young's modulus, respectively, were the strength of the applied process, especially in bone tissue engineering area. Contact angle measurements demonstrated about two times reduction in measured values, which shows improvement in hydrophilicity of PDA-modified PVA/PU-PANI fibers compared with PDA-coated PU-PANI ones. Swelling ratio and mass loss ratio calculations revealed enhancement in measured values as a function of homogeneous and dense coating, which arise from hydrophilicity of the polymeric substrate. The bioactivity test indicated that a dense layer of PDA strongly supports formations of hydroxyapatite-like crystals. Moreover, homogeneous decoration of conductive matrixes with PDA showed suitable cell viability, adhesion, and spreading while cell-scaffolds interactions improved under electrical stimulation. Higher expression of alkaline phosphatase and secretion of Collagen I under the electrical field proved the applicability of modified electroconductive scaffolds for further preclinical and clinical studies to introduce as a reconstructive bone substitute.

Competition of carrier bioresorption and drug release kinetics of vancomycin-loaded silicate macroporous microspheres to determine cell biocompatibility

Bioceramic porous microspheres are promising substances for dental and orthopedic bone void filling, tissue engineering and drug delivery applications. In this research, the structure and cytocompatibility of bioactive magnesium-calcium silicate macroporous microspheres loaded with vancomycin hydrochloride, an antibiotic drug, were studied. In this regard, bredigite (Ca7MgSi4O16), akermanite (Ca2MgSi2O7) and diopside (CaMgSi2O6) carriers were fabricated through a sequence of sol-gel, calcination, droplet extrusion and sintering processes, followed by impregnated with vancomycin. X-ray diffraction (XRD) and scanning electron microscopy verified the formation of the desired ceramic crystalline phases and macroporous characteristics of the carriers, respectively. Based on the MTT assay, the antibiotic-loaded bredigite, akermanite and diopside devices comparatively exhibited the lowest, intermediate and highest levels of human bone marrow mesenchymal stem cells (hBM-MSCs) viability and proliferation, respectively. It was concluded that the role of the carrier bioresorption kinetics prevails over the drug delivery kinetics in determining the cell biocompatibility of the devices.

Electroactive barium titanate coated titanium scaffold improves osteogenesis and osseointegration with low-intensity pulsed ultrasound for large segmental bone defects

For large segmental bone defects, porous titanium scaffolds have some advantages, however, they lack electrical activity which hinders their further use. In this study, a barium titanate (BaTiO3) piezoelectric ceramic was used to modify the surface of a porous Ti6Al4V scaffold (pTi), which was characterized by scanning electron microscopy, energy dispersive spectroscopy, X-ray photoelectron spectroscopy, and roughness and water contact angle analyses. Low intensity pulsed ultrasound (LIPUS) was applied in vitro and in vivo study. The activity of bone marrow mesenchymal stem cells, including adhesion, proliferation, and gene expression, was significantly superior in the BaTiO3/pTi, pTi + LIPUS, and BaTiO3/pTi + LIPUS groups than in the pTi group. The activity was also higher in the BaTiO3/pTi + LIPUS group than in the BaTiO3/pTi and pTi + LIPUS groups. Additionally, micro-computed tomography, the mineral apposition rate, histomorphology, and the peak pull-out load showed that these scaffold conditions significantly enhanced osteogenesis and osseointegration 6 and 12 weeks after implantation in large segmental bone defects in the radius of rabbits compared with those resulting from the pTi condition. Consequently, the improved osteogenesis and osseointegration make the BaTiO3/pTi + LIPUS a promising method to promote bone regeneration in large segmental bone defects for clinical application.

Native extracellular matrix, synthesized ex vivo by bone marrow or adipose stromal cells, faithfully directs mesenchymal stem cell differentiation

Mesenchymal stem cells (MSCs) are highly responsive to cues in the microenvironment (niche) that must be recapitulated ex vivo to study their authentic behavior. In this study, we hypothesized that native bone marrow (BM)- and adipose (AD)-derived extracellular matrices (ECM) were unique in their ability to control MSC behavior. To test this, we compared proliferation and differentiation of bone marrow (BM)-derived MSCs when maintained on native decellularized ECM produced by BM versus AD stromal cells (i.e. BM- versus AD-ECM).We found that both ECMs contained similar types of collagens but differed in the relative abundance of each. Type VI collagen was the most abundant (≈60% of the total collagen present), while type I was the next most abundant at ≈30%. These two types of collagen were found in nearly equal proportions in both ECMs. In contrast, type XII collagen was almost exclusively found in AD-ECM, while types IV and V were only found in BM-ECM. Physically and mechanically, BM-ECM was rougher and stiffer, but less adhesive, than AD-ECM.During 14 days in culture, both ECMs supported BM-MSC proliferation better than tissue culture plastic (TCP), although MSC-related surface marker expression remained relatively high on all three culture surfaces. BM-MSCs cultured in osteogenic (OS) differentiation media on BM-ECM displayed a significant increase in calcium deposition in the matrix, indicative of osteogenesis, while BM-MSCs cultured on AD-ECM in the presence of adipogenic (AP) differentiation media showed a significant increase in Oil Red O staining, indicative of adipogenesis. Further, culture on BM-ECM significantly increased BM-MSC-responsiveness to rhBMP-2 (an osteogenic inducer), while culture on AD-ECM enhanced responsiveness to rosiglitazone (an adipogenic inducer).These findings support our hypothesis and indicate that BM- and AD-ECMs retain unique elements, characteristic of their tissue-specific microenvironment (niche), which promote retention of MSC differentiation state (i.e. “stemness”) during expansion and direct cell response to lineage-specific inducers. This study provides a new paradigm for precisely controlling MSC fate to a desired cell lineage for tissue-specific cell-based therapies.

Pectin from Brassica oleracea var. italica triggers immunomodulating effects in vivo

The immunomodulatory ability of pectins is related with structural features such as the degree of methyl-esterification and branching, as well as the molecular mass. The pectin FB, extracted from broccoli stalks (Brassica oleracea var. italica) had low molecular mass, 56% methyl-esterification and galactose as the main neutral sugar, sharing some characteristics with the modified citrus pectin (MCP), which has been extensively studied in vivo and in vitro due its immunomodulator potential. Considering that broccoli has an important role in the diet, the main objective of this study was to investigate the ability of FB in modulating the immune system in vivo. At concentrations 100–500 μg/mL, FB did not affect the viability of macrophages. Evaluations on morphology and phagocytic activity showed that FB (500 μg/mL) increased the number of activated macrophages by 39% and phagocytic activity by 30% within 48 h. FB (200 mg/kg) administered intraperitoneally increased the number of peritoneal macrophages in mice by 490% after 24 h and modulated these cells for an activated phenotype. In mice, oral administration of FB (200 mg/kg) stimulated lymphocytes from spleen and bone marrow proliferation. FB did not induce nitric oxide (NO) production by macrophages and also did not affect the levels of pro-inflammatory interleukins IL-1β and IL-12 by peritoneal macrophages, but induced the production of the anti-inflammatory interleukin IL-10. The results could suggest that the anti-inflammatory effects triggered by FB could be related to its degree of esterification and pointed this polysaccharide as a target for the development of new immunomodulatory drugs.

3D-printed Ti6Al4V scaffolds coated with freeze-dried platelet-rich plasma as bioactive interface for enhancing osseointegration in osteoporosis

The serious complications after arthroplasty in patients with osteoporosis is a challenge for orthopedics. Three-dimensional (3D)-printed porous titanium (pTi) alloys have the character of low stiffness, similar to cortical bone, and are seemed as promising materials in orthopedic applications, but limited by the bio-inert of titanium alloy. Thus, improving the bioactivity of the prosthesis and bone regeneration around the implant are crucial for the osseointegration. Herein, we established rabbit model of osteoporosis by bilateral ovariectomy, and the bioactive interface was constructed by coating 3D-printed pTi scaffolds with autologous platelet-rich plasma (PRP) prepared by traditional and Freeze-Dried methods. The pTi have advantages of ideal pore size, porosity and mechanical properties matching with bone tissue. In vitro, PRP-coated pTi significantly promoted bone marrow mesenchymal stem cells (BMSCs) attachment, proliferation, migration, and osteogenic differentiation. In vivo studies of the implant coatings were performed using osteoporosis models. The results indicated that this PRP-coated bioactive interface enhanced bone regeneration and osseointegration. Besides, compared with Traditional PRP, the Freeze-Dried PRP-coated pTi exhibited the superiority of promoting cell activity and osteogenic differentiation of BMSCs, and better bone formation. The novel Freeze-Dried PRP-coated pTi shows potential for the development of artificial prosthesis with bioactive interface for patients with osteoporosis.

High-fat diet selectively decreases bone marrow lin- /CD117+ cell population in aging mice through increased ROS production.

Bone marrow (BM) stem cells (BMSCs) are an important source for cell therapy. The outcome of cell therapy could be ultimately associated with the number and function of donor BMSCs. The present study was to evaluate the effect of long-term high-fat diet (HFD) on the population of BMSCs and the role of reactive oxygen species (ROS) in aging mice. Forty-week-old male C57BL/6 mice were fed with HFD for 3 months with regular diet as control. Experiments were repeated when ROS production was reduced in mice treated with N-acetylcysteine (NAC) or using mice overexpressing antioxidant enzyme network (AON) of superoxide dismutase (SOD)1, SOD3, and glutathione peroxidase. BM and blood cells were analyzed with flowcytometry for lineage negative (lin- ) and Sca-1+ , or lin- /CD117+ , or lin- /CD133+ cells. Lin- /CD117+ cell population was significantly decreased with increased intracellular ROS and apoptosis and decreased proliferation in BM, not in blood, in HFD-treated mice without change for Sca-1+ or CD133+ cell populations in BM or blood. NAC treatment or AON overexpression effectively prevented HFD-induced intracellular ROS production and reduction of BM lin- /CD117+ population. These data suggested that long-term HFD selectively decreased BM lin- /CD117+ cell population in aging mice through increased ROS production.

SUN-278 Isolated Hypothalamic Langerhans Cell Histiocytosis in an Adult Manifesting as Panhypopituitarism

Background: Langerhans Cell Histiocytosis (LCH) is a rare disease characterized by abnormal proliferation of bone marrow derived histiocytes. Although predominantly a childhood disease, LCH can occur at any age and has an incidence of one to two cases per million in adults. LCH can involve a single organ or present as disseminated disease. Sites most commonly affected are bone, skin, bone marrow and pituitary. Isolated hypothalamic LCH is a rare entity. A few reports of LCH presenting as a hypothalamic mass exist but mainly in children. Treatment of adult LCH is derived from pediatric studies, and no standard of care exists. We report a unique case of an adult with panhypopituitarism secondary to a hypothalamic mass found to be biopsy proven isolated LCH treated with Vemurafenib, a BRAF V600E inhibitor. Clinical Case: A 66-year-old previously healthy man presented with progressively increasing confusion, cognitive decline and generalized weakness requiring hospital admission. Laboratory evaluation revealed an 8AM cortisol of 2.5 ug/dL (reference [ref] 7.0–25.0 ug/dL), ACTH of 9.3 pg/mL (ref 8–42 pg/mL), TSH of 0.106 uIU/mL (ref 0.35–5.5 uIU/mL), free T4 of 0.67 ug/dL (ref 0.7–1.25 ng/dL), LH <0.2 IU/L (ref 0.5–11 IU/L), FSH 0.6 IU/mL (ref 1–12 IU/L), total testosterone <12 ng/dL (ref 193 - 824 ng/dL), free testosterone <3.3 pg mL (ref 41.7 - 180.2 pg/mL), IGF1 41 ng/mL (ref 33–220 ng/mL) and prolactin of 31.5 ng/mL (ref 3.5–15 ng/mL). Findings were consistent with panhypopituitarism, and he was started on glucocorticoid and levothyroxine replacement therapy. MRI pituitary revealed an 18.1 x 13.8 x 15.8 mm hypothalamic mass with signal intensity alteration in the optic pathway. Patient developed polyuria with a serum sodium of 148 mmol/L (ref 136–145 mmol/L), elevated serum osmolality and low urine osmolality consistent with central DI and was started on oral DDAVP.The patient underwent right sided neuroendoscopic intraventricular biopsy of hypothalamic mass. Pathology was consistent with LCH with immunohistochemistry staining for CD1a, Langerin and BRAFV600E, with occasional cells staining for CD68, CD163 and S100. Bone marrow biopsy was normal. A NM PET scan revealed a hypermetabolic hypothalamic mass compatible with LCH without evidence of involvement of other sites including the skeletal system. Given histopathologically confirmed BRAF-mutated LCH, treatment with Vemurafenib was initiated. Behavioral changes gradually improved, and repeat MRI brain three months after treatment revealed decrease in size of hypothalamic mass. Conclusion: Isolated hypothalamic LCH is exceedingly rare. No clear guidelines exist for treatment of LCH in adults. BRAFV600E mutations are found in more than fifty percent of LCH cases and are associated with worse outcomes. This case demonstrates the successful use of targeted BRAF inhibitor therapy in an adult patient with BRAF mutated hypothalamic LCH.

Nonclassical Monocytes Sense Hypoxia, Regulate Pulmonary Vascular Remodeling, and Promote Pulmonary Hypertension.

An increasing body of evidence suggests that bone marrow-derived myeloid cells play a critical role in the pathophysiology of pulmonary hypertension (PH). However, the true requirement for myeloid cells in PH development has not been demonstrated, and a specific disease-promoting myeloid cell population has not been identified. Using bone marrow chimeras, lineage labeling, and proliferation studies, we determined that, in murine hypoxia-induced PH, Ly6Clo nonclassical monocytes are recruited to small pulmonary arteries and differentiate into pulmonary interstitial macrophages. Accumulation of these nonclassical monocyte-derived pulmonary interstitial macrophages around pulmonary vasculature is associated with increased muscularization of small pulmonary arteries and disease severity. To determine if the sensing of hypoxia by nonclassical monocytes contributes to the development of PH, mice lacking expression of hypoxia-inducible factor-1α in the Ly6Clo monocyte lineage were exposed to hypoxia. In these mice, vascular remodeling and PH severity were significantly reduced. Transcriptome analyses suggest that the Ly6Clo monocyte lineage regulates PH through complement, phagocytosis, Ag presentation, and chemokine/cytokine pathways. Consistent with these murine findings, relative to controls, lungs from pulmonary arterial hypertension patients displayed a significant increase in the frequency of nonclassical monocytes. Taken together, these findings show that, in response to hypoxia, nonclassical monocytes in the lung sense hypoxia, infiltrate small pulmonary arteries, and promote vascular remodeling and development of PH. Our results demonstrate that myeloid cells, specifically cells of the nonclassical monocyte lineage, play a direct role in the pathogenesis of PH.

T‐cell blast crisis of chronic myelogenous leukemia presented with coexisting p210 and p190 BCR‐ABL transcripts and t(10;11)(q11;p15)

BackgroundBlast transformation of chronic myelogenous leukemia (CML) to T lymphoblastic lymphoma/acute lymphoblastic leukemia (T‐LBL/ALL) is rare, and the molecular mechanism is still unclear.Case reportA 28‐year‐old woman who developed T‐ALL with coexpressing both p210 and p190 BCR‐ABL transcripts five years after the initial diagnosis of CML in chronic phase. The proliferation of bone marrow was extremely active with blast cells over 20%. Chromosome analysis revealed t(9;22)(q34;q11) and t(10;11)(q25;p15). Flow immunophenotyping showed that blasts expressed CD4, CD7, CD11b, CD38, CD34, CD33, and cCD3.ConclusionIt is the first T‐cell blast of CML case with coexisting p210 and p190 as well as additional chromosome translocations. Through review this case and previous reports, we will reveal that CML patients with T‐lymphocyte transformation depend on potential molecular and pathological mechanism.

Preparation and Characterization of Akermanite/Merwinite Scaffolds for Bone Tissue Repair

The ceramics in the system CaO–MgO–SiO2 has recently attracted a great deal of attention because they display a good in vitro bioactivity and have potential use as bone implants. Biphasic calcium-magnesium-silicate ceramics were prepared by a sol-gel method. The dried gel with chemical composition 3CaO.MgO.2SiO2 was thermally treated at 1200 °C for 2 hrs. The structural behavior of the synthesized ceramics was examined by means of X-ray diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR) and Scanning Electron Microscopy (SEM). Merwinite crystalline phase and akermanite phase were recognized. Then, porous akermanite/merwinite scaffolds were prepared to utilize polymer sponge method and evaluated by employing SEM. Furthermore, bone marrow stromal cells (BMSC) adhesion and proliferation on the scaffolds were evaluated by MTT assay test. Differentiation of the cells was assessed by measuring alkaline phosphatase (ALP) activity. The results demonstrated that BMSC adhered and spread well on akermanite scaffolds and proliferated with the increase in the culture time, and the differentiation rate of osteoblasts on scaffolds was comparable to that on blank culture plate control. Thus, the obtained results presented that the akermanite/merwinite scaffolds deserve attention for bone tissue engineering applications.