Proliferative Disorders Research Articles

Abstract B106: Antibody-drug conjugates targeted at human aspartyl (asparaginyl) β-hydroxylase

Abstract Antibody-drug conjugates (ADCs) consist of an antibody that is specific for a disease antigen linked to one or more drug molecules. The goal is to utilize the antibody as a means of specific delivery of the drug to diseased cells. Theoretically this should decrease systemic exposure to the drug and allow for use of a lower dose. In cancer, the approach employs an antibody targeted at a tumor-associated antigen and a cytotoxic or cytostatic drug. To date there have been three ADCs approved by the US FDA for cancer treatment, and many more are now in the development stage. The efficacy of any ADC is dependent on three factors: the antibody, the attached drug, and the linker used to connect them. While each of these factors is of import, with the growth of interest in ADCs for cancer therapy, the use of specific drugs and linkers has become more routine. Thus, the specificities of the antibody and target antigen are now perhaps the primary obstacles towards the development of novel ADCs. Human aspartyl (asparaginyl) β-hydroxylase (HAAH) is an embryonic/developmental protein, which is downregulated in normal cells after birth but overexpressed on the surface of many malignant cells. It has been demonstrated to be sufficient to induce cancer cell proliferation, motility, and invasiveness. The enzyme hydroxylates important residues in the NOTCH protein altering signal transduction pathways that lead to increased growth and metastatic potential. Increased levels of HAAH have been detected by immunohistochemistry in a diverse array of solid and hematologic cancers (n > 20), including liver, bile duct, brain, breast, colon, prostate, ovary, pancreas, and lung cancers as well as various leukemias. HAAH is not found in measurable quantities in normal tissue (n > 500), including normal adjacent tissue within cancer biopsy specimens or in benign proliferative disorders. We have developed a fully human antibody against HAAH, PAN-622,that displays exquisite specificity for cancer. Here we explore PAN-622 drug conjugates for ultimate use in treating both hematologic and solid tumors. PAN-622 was conjugated to three different drugs: a maytansinoid (DM1), monomethyl auristatin E (MMAE), and duocarmycin (DUO). DM1 was conjugated via a noncleavable thio-ether linker while MMAE and DUO were conjugated using valine-citruline containing linkers that are cleavable by cathepsin B in the endosomal compartment. Conjugation of drugs to the PAN-622 antibody had little effect on the affinity of the antibody for HAAH. Binding affinities were determined by immunoassay on fixed cancer cells (H460, human lung cancer line) and were ~0.1, 0.2, and 0.5nM for PAN-622-DM1, PAN-622-MMAE, and PAN-622-DUO, respectively. The affinity of PAN-622 for HAAH as expressed on live cells has previously been shown to be ~1nM. Efficacies of the three ADCs for killing of H460 cells were determined using an MTS assay. The PAN-622-DM1 had an EC50 of ~15nM. The EC50 for PAN-622-MMAE was ~60nM, and that for PAN-622-DUO was ~300nM. Importantly, both unconjugated PAN-622 and a nonrelevant antibody conjugated to MMAE did not display any killing of the H460 cell line. Efficacy was also measured on a representative hematologic cancer line, MOLM-14 (acute myelogenous leukemia) where EC50s were in the 20-50nM range for all three ADCs. This work serves as a proof of concept, laying the groundwork for further development of HAAH-targeted ADCs. Citation Format: Kanam Malhotra, Susan Walker, Chen Fang, Karamveer Birthare, Michael S. Lebowitz, Steven Fuller, Muctarr Sesay, Hossein A. Ghanbari. Antibody-drug conjugates targeted at human aspartyl (asparaginyl) β-hydroxylase [abstract]. In: Proceedings of the AACR-NCI-EORTC International Conference: Molecular Targets and Cancer Therapeutics; 2017 Oct 26-30; Philadelphia, PA. Philadelphia (PA): AACR; Mol Cancer Ther 2018;17(1 Suppl):Abstract nr B106.

Ectopic gestational trophoblastic disease: A 20-Year histopathological review in a tertiary center

Background: Gestational trophoblastic disease (GTD) is a group of proliferative disorders of the placental trophoblast. While the etiopathogenesis is not fully understood, this spectrum of diseases has varied histological appearances, with clinical behaviors ranging from benign to malignant. The burden of this group of lesions seems to be more in the developing countries. A high index of clinical suspicion, early laboratory confirmation of the diagnosis, and prompt treatment ensure total cures, even of malignant disease. These diseases, just like gestation itself, are known to occur in both intrauterine and extrauterine (ectopic) sites. Aim and Objectives: This study sought to determine the morphological pattern as well as the age and site distribution of the various forms of ectopic GTD histologically diagnosed in the University of Benin Teaching Hospital (UBTH), Benin City, Nigeria, between January 1993 and December 2012. It is a hospital-based, retrospective review utilizing materials from the archives of the Department of Pathology, UBTH. The parameters studied include the specific histological diagnoses and the age and site distribution. Results: A total of 28 cases of ectopic GTDs were encountered. There were 17 cases (60.7%) and 8 cases (28.6%) of GTD found in the right and left fallopian tubes, respectively. Other sites of ectopic GTD were the left ovary (2 cases; 7.1%) and the right ovary (1 case; 3.5%). Partial mole was the most common (75.0%), followed by complete mole and invasive mole (10.7% each), and then choriocarcinoma (3.6%). The malignant: benign ratio was 1:27. Conclusion: Ectopic GTD, though still a relatively uncommon occurrence, should not be overlooked as a possibility in women with ectopic gestation, hence the need for histopathological and other ancillary evaluation of all ectopic conceptuses.

EBV in T-/NK-Cell Tumorigenesis.

Epstein-Barr virus (EBV), which is associated with B-cell proliferative disorders, also transforms T- or natural killer (NK)-lineage cells and has been connected with various T- or NK (T/NK)-cell malignancies, such as extranodal NK/T-cell lymphoma-nasal type and aggressive NK-cell leukemia. Chronic active EBV (CAEBV) disease , which occurs most often in children and young adults in East Asia, is an EBV-associated T-/NK-cell lymphoproliferative disease. Patients with CAEBV often progress to overt lymphoma or leukemia over a long-term clinical course. EBV's transforming capacity in B cells is well characterized, but the molecular pathogenesis of clonal expansion caused by EBV in T/NK cells has not yet been clarified. In the primary infection, EBV infects B cells and epithelial cells and may also infect some T/NK cells. In some individuals, because of poor presentation by specific human leukocyte antigens or the genetic background, EBV-infected T/NK cells evade host immunity and survive. Occasionally, with the help of viral oncogenes, EBV-associated T/NK lymphoproliferative diseases, such as CAEBV, may develop. The subsequent accumulation of genetic mutations and/or epigenetic modifications in driver genes, such as DDX3X and TP53, may lead to overt lymphoma and leukemia. Activation-induced cytidine deaminase and the APOBEC3 family, driven by EBV infection, may induce chromosomal recombination and somatic mutations.

Rosai-Dorfman Disease (Sinus Histiocytosis With Massive Lymphadenopathy): A Case Report

Sinus Histiocytosis with Massive Lymphadenopathy also known as Rosai-Dorfman Disease is a rare histiocytic proliferative disorder of unknown etiology. It presents as massive bilateral painless cervical lymphadenopathy. We report a case of RosaiDorfman disease in a 11 year old boy who presented with bilateral painless cervical lymohadenopathy. Only a few such cases have been reported in the literature hence this case is being reported for its rarity.

Rosai-Dorfman Disease: Rare Pulmonary Involvement Mimicking Pulmonary Langerhans Cell Histiocytosis and Review of the Literature

Rosai-Dorfman disease (RDD), also known as sinus histiocytosis with massive lymphadenopathy, is a rare nonmalignant lymphohistiocytic proliferative disorder. We report a patient with RDD who presented with multiple skin lesions, pulmonary involvement, and CT manifestations mimicking Langerhans cell histiocytosis, which improved after initiation of corticosteroid treatment.

Anti-vascular endothelial growth factor treatment for retinopathy of prematurity

Retinopathy of prematurity (ROP) is a proliferative disorder of the developing retina in premature and low birth weight infants. Recently, the role of vascular endothelial growth factor (VEGF) in the pathophysiology of ROP has been well studied and anti-VEGF drugs have been used in phase 2 to treat ROP patients in many ways. At first, ophthalmologists began to give intravitreal bevacizumab (IVB) or ranibizumab off-label to treat ROP as a salvage treatment after failure in laser photocoagulation or in combination with laser as an adjuvant treatment for patients had media opacity or rigid pupil. Now anti-VEGF drugs are also used as monotherapy in type I ROP or perioperative use in stage 4/5 ROP. Questions remain regarding long-term safety, dose, timing, visual outcomes and long-term effects, including systemically.

WITHDRAWN: Targeting mTORC1/2 with OSI-027 inhibits proliferation and migration of keloid keratinocytes

// Jun Chen 1, * , Ke Liu 1, * , Yang Liu 1 , Xue Wang 1 and Zhen Zhang 1 1 Department of Dermatology and Dermatologic Surgery, Shanghai Ninth People’s Hospital, Affiliated to Shanghai Jiaotong University School of Medicine, Center for Specialty Strategy Research of Shanghai JiaoTong University China Hospital Development Institute, Shanghai 200011, China * These authors contributed equally to this work Correspondence to: Zhen Zhang, email: zz6503@126.com Keywords: keloid keratinocytes; cell migration; mTOR; OSI-027; signaling Received: August 08, 2017 Accepted: December 11, 2017 Published: January 02, 2018 ABSTRACT Keloid is a dermal proliferative disorder characterized by the excessive keratinocyte proliferation and migration. mTOR over-activation is involved in the process. Here, we show that both mTOR complex 1 (mTORC1) and mTORC2 were hyper-activated in keloid-derived primary keratinocytes. OSI-027, an mTOR kinase inhibitor, potently inhibited proliferation and migration of keloid keratinocytes. OSI-027 disrupted the assembly of mTORC1 (mTOR-Raptor) and mTORC2 (mTOR-Rictor-mLST8). Further, OSI-027 completely blocked the phosphorylations of the mTORC1 substrates (S6K1, S6 and 4EBP1) and the mTORC2 substrate (AKT, at Ser-473). OSI-027 was potent than rapamycin in inhibiting keloid keratinocytes. Moreover, restoring mTORC1 activation by the introduction of the constitutively active S6K1 only partly alleviated OSI-027-induced suppression on keloid keratinocytes. Notably, mTOR2 inhibition by Rictor siRNA also inhibited keloid keratinocyte proliferation and migration, although less efficiently than OSI-027. Together, concurrent targeting of mTORC1/2 by OSI-027 potently inhibits keloid keratinocyte proliferation and migration.

Abstract 36: Preclinical models of multiple myeloma

Abstract Multiple myeloma is a clonal B-cell malignancy characterized by the accumulation of terminally differentiated, antibody-producing plasma cells in the bone marrow. Genetic mutations within the myeloma cells and their interaction with various cytokines and growth factors contribute to the invasiveness of the disease and enhanced drug resistance. Patients are generally asymptomatic until very late-state disease. However, once disease is present, localization in the bones, especially the spine, is common. There are few preclinical models that recapitulate human disease. In this work, we have evaluated the human MM.1S model and the murine 5TGM-1 model. MM.1S was derived from a 42-year-old African American woman and has been documented to express CD25, CD38, CD52, and CD59. It also expresses the glucocorticoid receptor and is dexamethasone sensitive. To more effectively monitor in vivo disease progression, we transfected the MM.1S line with luciferase (MM.1S-pMMP-LucNeo). With bioluminescence imaging (BLI) we track and monitor disseminated disease progression over time and find reproducible in vivo growth in SCID beige mice. The median tumor volume doubling time is ~2 days and median time to morbidity/mortality endpoint is 35-45 days. MM.1S-pMMP-LucNeo was also evaluated in NSG mice which show more aggressive disease onset (staging 7 days post implant vs. 15 days in SCID mice). In NSG mice the tumor volume doubling time is ~1.8 days and median time to endpoint is ~21 days, roughly half the overall survival time observed in the SCID mouse. In all mouse strains we have investigated, the animals are relatively asymptomatic throughout the lifetime of the study and only begin to present symptoms at late-stage disease, similar to how the disease manifests itself in humans. Common, late-stage clinical signs in the mice include lethargy and paralysis. BLI on these animals shows localization of the signal in the spine, long bones, and mandible. As in the clinic, bortezomib (Velcade®) has limited effect on disease progression in this model. Expression of CD138 is a hallmark of plasma cells and multiple myeloma cells. In the NSG study we determined, by flow cytometry, that >85% of the CD45- gate in the bone marrow was CD138+ 21 days post-implant, suggesting significant engraftment of the tumor cells within the bone marrow. 5TGM-1 is a syngeneic model generated from the C57BL/KaLwRij mouse which is predisposed to develop several monoclonal B-cell proliferative disorders, including a low percentage (0.5%) of the mice developing multiple myeloma. From this strain, several cell clones were isolated and called the 5TMM series, which included the 5T33MM clone. 5TGM-1 is a sub-clone of 5T33MM cells and we have used 5TGM-1-luc to monitor disease progression and localization in vivo. The 5TGM-1-luc model is aggressive and BLI has shown localization of disease in the long bones. We have compared this model in both the syngeneic C57BL/KaLwRij strain and the immune-deficient NIH III (triple deficient) strain. Disease progression behaves similarly in both strains of mice, with significant tumor cells localizing in the spine and the long bones. The model is sensitive to traditional chemotherapies like doxorubicin and cyclophosphamide, but bortezomib (Velcade) only produces modest activity and carfilzomib has shown little activity at the dose and schedule tested. A greater understanding of the preclinical behavior of these models, including response to standard of care and molecular profiling, will provide critical insight into designing more clinically relevant preclinical experiments. Citation Format: Erin Trachet, Maryland Rosenfeld Franklin. Preclinical models of multiple myeloma [abstract]. In: Proceedings of the Second AACR Conference on Hematologic Malignancies: Translating Discoveries to Novel Therapies; May 6-9, 2017; Boston, MA. Philadelphia (PA): AACR; Clin Cancer Res 2017;23(24_Suppl):Abstract nr 36.

Nattokinase: an updated critical review on challenges and perspectives.

Natto, a fermented soybean food, has been consumed by oriental people for more than 1000 years. Nattokinase, formerly called subtilisin NAT is a well studied protease of microbial origin that possesses fibrinolytic (anti-clotting) activities. Due to its strong fibrinolytic and thrombolytic activity, Nattokinase is regarded as a precious dietary supplement or nutraceutical for the oral thrombolytic therapy. Nattokinase is witnessed to be a useful enzyme for the com-plete removal of the vitreous and associated proliferative tissues in proliferative vitreo retinal disorders. This review focuses on the native and recombinant Nattokinase from bacteria and other sources, their production, purification, immobilization and nano-immobilization studies, which aid in ameliorating their properties to suit the targeted industrial applications. Recent development in these fields are presented and discussed, and prospective developments are suggested.

Castleman Disease and Rosai-Dorfman Disease.

This chapter describes the main features of two different diseases, Castleman Disease (CD) and Rosai-Dorfman Disease (RDD). Castleman disease (CD) is a clinical and histopathologically heterogeneous lymphoproliferative disorder that encompasses at least three distinct entities with some common overlapping morphological features: Hyaline Vascular CD (HVCD), Unicentric Plasma Cell CD and Multicentric CD. The most important feature of HVCD is the presence of abnormal germinal centers with hyaline-vascular transformation, sometimes showing multiple germinal centers within a single reactive lymphoid follicle, this outlining HVCD as a disorder of follicular dendritic cells. Unicentric and multicentric CD are, in contrast, lymphoproliferative lesions. Proinflammatory hypercytokinemia is an essential feature of multicentric CD, distinguished by a florid clinical presentation. Rosai-Dorfmann Disease is a histiocytic proliferative disorder diagnosed by the presence of tissue infiltration by S100-positive CD1a-negative histiocytes and plasma cell aggregates, often with Russell bodies. A typical, though not specific, characteristic of the disease is emperipolesis. Initially considered to be an inflammatory/reactive condition, molecular studies suggest that at least some cases of RDD could be considered as a low-grade histiocytic neoplastic process.

Video-Assisted Thoracoscopic Thymectomy Langerhans Cell Histiocytosis

Introduction: Langerhans cell histiocytosis (LCH) is an exceedingly rare proliferative disorder in which pathologic histiocytic cells accumulate in nearly every organ. Although the deposited cells share morphology and antigens with the skin Langerhans cell, the cell of origin is actually a myeloid dendritic cell. LCH is most common in the pediatric population from ages 1 to 3 effecting ~1–2 patients per million. The most common manifestation of the disease is lytic bone lesions accounting for 77% of those diagnosed. Lymph nodes and the lung are involved in 19% and 10% of cases, respectively. Methods: Our patient, a 5-month-old, 6-kg female with mild pulmonary valve stenosis, had both thymic and lung tissue involvement. To date, there has never been a report of a thymic LCH with lung metastases in an infant. Results: She underwent an echocardiogram to assess the degree of her pulmonary stenosis, which incidentally showed a large thymic cyst. That cyst was further characterized with CT scan as a heterogenous mass with multiple lung nodules. The patient was meeting all milestones appropriately and asymptomatic without cardiac symptoms, respiratory difficulty, or feeding issues. To rule out a thymoma, the patient underwent right video-assisted thoracoscopic thymectomy with wedge biopsy of the right lower lobe of the lung. Conclusion: Final pathology report showed LCH in both tissue samples. The patient did well without complications and was discharged home expeditiously. She is now under the care of the medical oncologists for continued management. No competing financial interests exist. Runtime of video: 4 mins 30 secs

Herpes Simplex Virus and Langerhans Cell Histiocytosis

Langerhans cell histiocytosis (LCH) is a rare histiocytic proliferative disorder of unknown etiology and mainly affects young children. The histological feature is granuloma-like proliferation of langerhans-type dendritic cells. Although the possible role of viruses such as Epstein-Barr virus (EBV, Human Herpes virus -4), Human Herpes virus-6 (HHV-6), Herpes Simplex virus (HSV) types 1 and 2 and Cytomegalovirus (CMV, Human Herpes virus-5) is suggested in the pathogenesis of LCH by some investigators, its exact pathophysiology has not been cleared yet. In this study, we investigated the presence of HSV types 1 and 2 in Iranian children with LCH. In this retrospective study, we investigated the prevalence of presence of HSV types 1 and 2 (in 30 patients with LCH), using paraffin-embedded tissue samples and 30 age and tissue-matched controls (operated for reasons other than infectious diseases) from the Department of Pediatric Pathology, Tehran, Iran, by nested Polymerase Chain reaction method. No ethical issues arose in the study, because only the pathology reports were reviewed and patients were anonymous. We failed to find HSV types 1 and 2 DNA in any of the 30 patients with LCH or the control group. According to our findings, HSV types 1 and 2 do not appear to have any etiologic role in the pathogenesis of LCH in Iranian children. These results are in accordance with previous investigations with negative findings.

Odorant Receptor 51E2 Agonist β-ionone Regulates RPE Cell Migration and Proliferation.

The odorant receptor 51E2 (OR51E2), which is well-characterized in prostate cancer cells and epidermal pigment cells, was identified for the first time as the most highly expressed OR in human fetal and adult retinal pigment epithelial (RPE) cells. Immunofluorescence staining and Western blot analysis revealed OR51E2 localization throughout the cytosol and in the plasma membrane. Additionally, immunohistochemical staining of diverse layers of the eye showed that the expression of OR51E2 is restricted to the pigment cells of the RPE and choroid. The results of Ca2+-imaging experiments demonstrate that activation of OR51E2 triggers a Ca2+ dependent signal pathway in RPE cells. Downstream signaling of OR51E2 involves the activation of adenylyl cyclase, ERK1/2 and AKT. The activity of these protein kinases likely accounts for the demonstrated increase in the migration and proliferation of RPE cells upon stimulation with the OR51E2 ligand β-ionone. These findings suggest that OR51E2 is involved in the regulation of RPE cell growth. Thus, OR51E2 represents a potential target for the treatment of proliferative disorders.

MiR-548a-3p Promotes Keratinocyte Proliferation Targeting PPP3R1 after Being Induced by IL-22.

Psoriasis is an immune-mediated chronic skin disorder where T cells play a main role, and numerous inflammatory cytokines are implicated in its pathogenesis by initiating keratinocyte proliferation. Interleukin-22 (IL-22), an IL-10 family cytokines, is critical in the pathogenesis and development of psoriasis. To determine the target of microRNA (miR) -548a-3p and investigate its role in keratinocyte proliferation after treating human keratinocytes (HaCaT) with IL-22, we used quantitative reverse transcriptase PCR to measure the expression of miR-548a-3p in both HaCaT cells stimulated with IL-22 and psoriatic lesions, and then detected the biological function of miR-548a-3p in HaCaT cells by performing Counting Kit-8 (CCK-8) assays. Luciferase reporter assay was conducted to determine the target gene of miR-548a-3p. Immunohistochemistry and Western blot were performed to verify the target gene. Results showed that miR-548a-3p was significantly upregulated both in HaCaT cells treated with IL-22 and psoriatic lesions. The over expression of miR-548a-3p could promote the proliferation of HaCaT cells. Luciferase was mutated in the 3'UTR of PPP3R1, a gene coding Calcineurin. Immunohistochemistry and Western blot demonstrated that the expression of PPP3R1 decreased respectively in psoriatic lesions and HaCaT cells. In conclusion, the expression of miR-548a-3p is upregulated in IL-22 mediated keratinocyte proliferative disorder like psoriasis. The impact of miR-548a-3p on keratinocyte proliferation may be implemented by targeting PPP3R1 and T regulatory cells may be involved in the pathogenesis of psoriasis.

Carbon monoxide in lung cell physiology and disease.

Carbon monoxide (CO) is an endogenously produced gas that has gained recognition as a biological signal transduction effector with properties similar, but not identical, to that of nitric oxide (NO). CO, which binds primarily to heme iron, may activate the hemoprotein guanylate cyclase, although with lower potency than NO. Furthermore, CO can modulate the activities of several cellular signaling molecules such as p38 MAPK, ERK1/2, JNK, Akt, NF-κB, and others. Emerging studies suggest that mitochondria, the energy-generating organelle of cells, represent a key target of CO action in eukaryotes. Dose-dependent modulation of mitochondrial function by CO can result in alteration of mitochondrial membrane potential, mitochondrial reactive oxygen species production, release of proapoptotic and proinflammatory mediators, as well as the inhibition of respiration at high concentration. CO, through modulation of signaling pathways, can impact key biological processes including autophagy, mitochondrial biogenesis, programmed cell death (apoptosis), cellular proliferation, inflammation, and innate immune responses. Inhaled CO is widely known as an inhalation hazard due to its rapid complexation with hemoglobin, resulting in impaired oxygen delivery to tissues and hypoxemia. Despite systemic and cellular toxicity at high concentrations, CO has demonstrated cyto- and tissue-protective effects at low concentration in animal models of organ injury and disease. These include models of acute lung injury (e.g., hyperoxia, hypoxia, ischemia-reperfusion, mechanical ventilation, bleomycin) and sepsis. The success of CO as a candidate therapeutic in preclinical models suggests potential clinical application in inflammatory and proliferative disorders, which is currently under evaluation in clinical trials.

Pharmacological and transcriptional inhibition of the G9a histone methyltransferase suppresses proliferation and modulates redox homeostasis in human microvascular endothelial cells

Epigenetic mechanisms, including histone post-translational modifications, are central regulators of cell cycle control. The euchromatic G9a histone methyltransferase (G9a HMT) is a key enzyme catalyzing histone H3 methylation on lysines 9 and 27, and its dysregulation has been linked to uncontrolled proliferation of tumor cells. Here, we have investigated the effect of G9a HMT silencing on cell proliferation of microvascular endothelial cells, a process necessary to sustain tumor growth through the formation of the vascular capillary network.Inhibition of G9a HMT activity in human microvascular endothelial cells (HMEC-1) was performed either pharmacologically, by treatment of cells with BIX-01294 or chaetocin, or transcriptionally, using shRNA. Cell viability and proliferation were examined using the resazurin reduction assay, flow cytometry and immunostaining of phosphorylated checkpoint kinase 1 (pSer317Chk1). Expression of cell cycle- and redox homeostasis-related genes was determined by quantitative PCR. Reactive oxygen species production was measured by oxidation of the fluorescent probe 2′,7′-dichlorodihydrofluorescein diacetate and the cell’s total antioxidant capacity by using the ABTS assay.Inhibition of G9a HMT activity by BIX-01294 treatment or by shRNA attenuated the proliferation of HMEC-1, nuclear localization of phosphorylated Chk1, and induced cell cycle arrest in G1 phase. Transcriptional analysis demonstrated increased gene expression of the cyclin-dependent kinase (CDK) inhibitor p21, and also of Rb1, in BIX-01294 treated cells. Decreased proliferation rate was accompanied by enhanced antioxidant potential of HMEC-1 cells, as demonstrated by reduced production of reactive oxygen species, increased total antioxidant capacity and expression of the antioxidant enzymes catalase and superoxide dismutase 1.Collectively, our results demonstrate of the central role of G9a HMT in the promotion of endothelial cells proliferation, and suggest that endothelial G9a HMT may be a target in the treatment of vascular proliferative disorders and tumor neovascularization.

Synovial Chondromatosis in the Temporomandibular Joint: Report of Two Cases

Background Synocial chondromatosis(SC), a proliferative disorder of the synovial membrane. The etiology or cause of SC remains unclear. SC usually occurs in large articular joints such as knee, hip, elbow, and ankle. SC of the TMJ is very rare. It is a benign disease that mainly affects unilateral side. It can form cartilagenous and calcified loose bodies of various sizes and cause abnormal function of TMJ. Case Report In this paper, we report two cases of SC in the upper joint space of the left TMJ. One complained that "Sometimes the left jaw joint feels disoriented" and the other had no symptoms. CT scan and MRI showed left TMJ space widening, multiple tiny calcified mass. After clinical and radiographic analysis, we performed surgical removal of the lesion under genereal anesthesia. In the histologic examination, synovial chondromatosis was diagnosed in both patients. Conclusions We report two cases of synovial chondromatosis in the upper joint space of the left TMJ. We performed surgical removal of the lesion. The two patients showed good prognosis without recurrence or pain up to date.

Monoclonal IgG in MGUS and multiple myeloma targets infectious pathogens.

Subsets of mature B cell neoplasms are linked to infection with intracellular pathogens such as Epstein-Barr virus (EBV), hepatitis C virus (HCV), or Helicobacter pylori. However, the association between infection and the immunoglobulin-secreting (Ig-secreting) B proliferative disorders remains largely unresolved. We investigated whether the monoclonal IgG (mc IgG) produced by patients diagnosed with monoclonal gammopathy of undetermined significance (MGUS) or multiple myeloma (MM) targets infectious pathogens. Antigen specificity of purified mc IgG from a large patient cohort (n = 244) was determined using a multiplex infectious-antigen array (MIAA), which screens for reactivity to purified antigens or lysates from 9 pathogens. Purified mc IgG from 23.4% of patients (57 of 244) specifically recognized 1 pathogen in the MIAA. EBV was the most frequent target (15.6%), with 36 of 38 mc IgGs recognizing EBV nuclear antigen-1 (EBNA-1). MM patients with EBNA-1-specific mc IgG (14.0%) showed substantially greater bone marrow plasma cell infiltration and higher β2-microglobulin and inflammation/infection-linked cytokine levels compared with other smoldering myeloma/MM patients. Five other pathogens were the targets of mc IgG: herpes virus simplex-1 (2.9%), varicella zoster virus (1.6%), cytomegalovirus (0.8%), hepatitis C virus (1.2%), and H. pylori (1.2%). We conclude that a dysregulated immune response to infection may underlie disease onset and/or progression of MGUS and MM for subsets of patients.

Hymenoptera-induced anaphylaxis: is it a mast cell driven hematological disorder?

Hymenoptera-induced allergy (HVA) is a common cause of anaphylaxis and may be fatal. It is associated with systemic mastocytosis in about 7% of adult patients. Systemic mastocytosis is a proliferative disorder of hematopoietic mast cell progenitors. There is recent data on the association between systemic mastocytosis and HVA. Hymenoptera venoms are the most common immunoglobulin E-mediated elicitors of anaphylaxis in patients with mastocytosis. Some patients with systemic mastocytosis do not have cutaneous involvement and have only been identified because of HVA. These patients do have distinct demographic, clinical, and laboratory features as compared with patients with mastocytosis with skin lesions. They show a male predominance, more severe (cardiovascular) as compared to skin symptoms, lower baseline serum tryptase, less bone marrow mast cell aggregates, and KIT mutations restricted to mast cell lineage. Although hymenoptera-induced anaphylaxis is not primarily a mast cell-driven hematological disorder, the latter is present in a significant proportion of patients and has to be excluded by basal serum tryptase determination, skin inspection as well as exclusion of systemic mastocytosis by D816V mutation analysis in peripheral blood and/or bone marrow examination in selected patients.

Epstein-Barr Virus Mucocutaneous Colonic Ulcer in an Immunocompetent Patient

Epstein Barr virus (EBV) positive mucocutaneous ulcers (MCU) were first described in 2010 in immunosuppressed individuals. Lesions have been noted in the oral mucosa, skin, and gastrointestinal tract. Although these lesions are typically benign and will heal with conservative therapy with reversal of immunosuppression, rare reports have described progression to disseminated Hodgkin's lymphoma (HL). We report a case of EBVMCU in an immunocompetent patient during a period of active EBV infectious mononucleosis superimposed on campylobacter colitis. A 47-year-old, previously healthy male presented to the hospital for evaluation for a 1-month history of bloody diarrhea associated with recent development of pharyngitis and flu-like symptoms. Physical exam noted pharyngeal erythema, symmetrical bilateral cervical lymphadenopathy with diffuse abdominal tenderness to palpation. Initial labs noted normal hemoglobin and WBC with a lymphocyte predominance. Transaminases were elevated with an AST 202 U/L, ALT 410 U/L and an alkaline phosphatase of 870 U/L. CT of his abdomen and pelvis revealed circumferential thickening of the rectum with inflammatory stranding, diffuse pelvic, abdominal, and thoracic lymphadenopathy, splenomegaly, and multiple hepatic lesions: collectively concerning for malignancy. Serum AFP, CEA, CA 19-9, and HIV markers were normal. Fecal calprotectin was elevated over 830ug/g. Hepatic MRI noted presence of cysts. Stool cultures were positive for campylobacter. He was started on azithromycin and after several days of therapy, symptoms did not improve. Colonoscopy revealed several clean based ulcers in the rectum and sigmoid-colon (Figure 1) with biopsies positive for EBVMCU (Figure 2). Serum EBV IgM was positive. He was ultimately discharged in stable condition with instructions for close follow-up. To date, 51 cases of EBVMCU have been described in the literature, 5 of which developed within the colon. All reports describe a predisposing factor of immunosuppression by either medication or age-related immunosenescence in patients >60. We report a presumed novel case in which colonic EBVMCU developed in a young immunocompetent patient during active EBV infection superimposed with campylobacter colitis. It is imperative to keep a broad differential and investigate persistent symptoms despite a diagnosis. EBVMCU falls on a spectrum of EBV-associated proliferative disorders, including HL, thus close surveillance is needed to ensure disease resolution.Figure: Colonoscopy acquired images depicting several clean based rectal ulcers.Figure: (Left) H&E stain of rectal ulcer noting colonic mucosa with dense lymphoplasmacyctic infiltrate. (Right) EBV in situ hybridization indicating presence of EBV DNA within the nuclei of infected cells.