Major Role In Disease Progression Research Articles

Advances in molecular evaluation of myeloproliferative neoplasms.

Myeloproliferative neoplasms (MPN) are a group of clonal hematopoietic stem cell disorders with uncontrolled proliferation of one or more hematopoietic cell types, including myeloid, erythroid and megakaryocytic lineages, and minimal defect in maturation. Most MPN are associated with well-defined molecular abnormalities involving genes that encode protein tyrosine kinases that lead to constitutive activation of the downstream signal transduction pathways and confer cells proliferative and survival advantage. Genome-wide sequencing analyses have discovered secondary cooperating mutations that are shared by most of the MPN subtypes as well as other myeloid neoplasms and play a major role in disease progression. Without appropriate management, the natural history of most MPN consists of an initial chronic phase and a terminal blast phase. Molecular aberrations involving protein tyrosine kinases have been used for the diagnosis, classification, detection of minimal/measurable residual disease, and target therapy. We review recent advances in molecular genetic aberrations in MPN with a focus on MPN associated with gene rearrangements or mutations involving tyrosine kinase pathways.

Concealed connection between COVID-19 infection and periodontitis: A survey in Jammu population

Introduction: On 11 March 2020, WHO has declared COVID-19 infection a worldwide pandemic as it had severely affected almost every country in the world? Many researchers have given cytokine storm as the basic pathogenesis for COVID 19 complications. Interestingly, Periodontal Disease (PD) is also an inflammatory disease and cytokines play a major role in disease progression. The aim of this survey is to assess the prevalence of gum diseases among COVID 19 patients prior to getting infection.Methodology: This patient’s self-assessed survey was created and administered a 32-item questionnaire for COVID positive patients. All questions were framed with the help of various standard national and international guidelines and information available at the time of survey.Result: Out of 400 questionnaire forms, we received 376 responses and out of 376 we excluded 74 partially filled responses. 73.2% had gum diseases prior to COVID-19 and 55% had mobile teeth prior to COVID-19.Conclusion: There can be definitely an impact of periodontal diseases on progression and complication of COVID-19 and vice-versa. Knowing Periodontal Disease and it’s pathogenic (inflammatory) link with COVID-19 infection would help in identifying the risk groups and establish apropos recommendation.

Metabolic Abnormalities Associated with T-Cell Exhaustion in CLL Eµ-TCL1 Murine Model

Introduction. T-cell differentiation and immune responses are associated with major metabolic changes that control cell fate upon antigen exposure. Chronic lymphocytic leukemia (CLL) is characterized by a marked T-cell dysfunction that plays a major role in disease progression and failure of adoptive cell therapy. A handful of recent studies have demonstrated mitochondrial and metabolic defects in the T-cell compartment in CLL patients. However, to date, no studies have been done to validate such metabolic abnormalities in the Eµ-TCL1 model, the most established and reliable murine model for studying CLL. Therefore, we hypothesized that CD8+ T-cells derived from the Eµ-TCL1 adoptive transfer model exhibit metabolic and mitochondrial disturbance that might contribute to the overall dysfunctional state of these cells. Methods. To investigate the influence of leukemic burden on the differentiation phenotypes, metabolic state and gene expression profile of CD8+ T-cells, C57BL/6 mice adoptively transferred with Eµ-TCL1 leukemic splenocytes were euthanized at varying time points. Once disease was established, spleens were collected for analysis. Mice were divided into 3 groups; early, intermediate, and late disease, based on the peripheral blood leukemic burden at endpoint. Multicolor flow cytometry was used for identification of T-cell phenotypes and measurement of mitochondrial mass, membrane potential, reactive oxygen species (ROS) and cellular glucose uptake. For T-cell activation studies, we cultured splenocytes in the presence of CD3/CD28 dynabeads and 30IU/ml hIL-2 for 72 hours. Nanostring metabolic pathways gene expression analysis was done on splenic CD8+ T-cells isolated from early and late-stage Eµ-TCL1 adoptive transfer mice via fluorescence-activated cell sorting. Differential gene expression was measured using ROSALIND cloud-based software platform. Results. Unsupervised t-distributed stochastic neighborhood embedding (tSNE) and Xshift clustering analyses revealed a gradual enrichment of specific CD8+ T-cell populations along disease progression. These populations upregulated exhaustion-related markers (EOMES, TOX, PD-1 and KLRG-1) and significantly downregulated TCF-1 expression, a transcription factor responsible for cell self-renewal capacity. Further analysis demonstrated a significant downregulation of PGC-1α expression, a master regulator of mitochondrial function, in Eµ-TCL1 CD8+ T-cells with high EOMES and low TCF-1 expression levels (EOMEShi TCF-1lo) compared to EOMEShi TCF-1hi cells. At late disease stage, we noticed significant mitochondrial depolarization, an increase in mitochondrial ROS and marked reduction in glucose uptake capacity in the Eµ-TCL1 CD8+ T-cells. Post in vitro activation, there was a significant increase in Eµ-TCL1 CD8+ T-cell mitochondrial biogenesis, membrane potential and ROS production compared to wild type (WT) controls. However, Eµ-TCL1 cultures showed significant enrichment in EOMEShi TCF-1lo CD8+ T-cell population, that was not detected in the activated WT cells. Interestingly, this population exhibited marked downregulation of PGC-1α, SOD-2 and NRF-2 expression compared to the populations maintaining TCF-1 expression. Moreover, we found that Eµ-TCL1 CD8+ T-cells had increased frequency of effector memory phenotypes compared to more central memory cells in WT cultures. These effector memory populations accumulated significant levels of mitochondrial ROS and depolarized mitochondria compared to the central memory counterparts. Nanostring analysis revealed disease stage-dependent changes in gene expression, where glycolysis was the top differentially expressed pathway in the late versus early-stage CD8+ T-cells as indicated by the gene set global significance score. On the other hand, negative significance scores were assigned to AMPK and autophagy pathways indicating a probable downregulation of these pathways during late phases of the disease. Conclusion. Our results demonstrate mitochondrial and metabolic abnormalities in Eµ-TCL1-derived CD8+ T-cells and point to possible defects in cellular stress signaling response that might lead to accumulation of dysfunctional mitochondria and push the cells towards exhausted states.

IMMU-20. MODULATION OF THE PERIPHERAL T CELL BIOLOGY IN GLIOBLASTOMA

Abstract Cancer is a systemic disease. Due to the exceedingly rare occurrence of metastasis of cerebral glioma, systemic alterations have, however, not been considered to play a major role in disease progression of glioma. T cells orchestrate the adaptive immune response in an antigen-specific, cytokine mediated manner. The aim of our study was to investigate how cerebral glioma impacts systemic T cell immunobiology. We performed gene expression profiling of peripheral blood T cells in patients with IDHwt glioblastomas as well as in a murine glioma model. In addition, we analyzed the levels of soluble immune biomarkers in patient blood and performed flow-cytometric phenotyping of human peripheral blood CD3+ T cells. We discovered a significant skewing of peripheral T cell phenotypes in IDHwt glioblastoma patients compared to healthy donors (HD), showing CD4+ TH1 expansion and reduced numbers of T follicular helper cells (TFH), TH1* and mucosa associated invariant T (MAIT) cells, while TH2 and TH17 percentages remained stable. Interestingly, peripheral memory CD4+ T cells exhibited reduced Fas and PD-1 expression, while CD8 T cells were primarily affected in the non-memory CD45RA+ compartment, displaying reduced numbers of CD8+ CD127+ CD27+ cells. Compared to healthy individuals, GBM patients had significantly increased levels of soluble CD27 while levels of soluble CD25 were reduced (p < 0.05). GSEA and ORA analysis of differentially expressed genes in murine gliomas showed alterations in RNA binding and processing, as well as ribosomal activity in both cell types, indicating systemic modulation in translational- and cell cycle pathways in glioblastoma. Taken together, our results demonstrate a significant skewing of the peripheral T immunobiology in patients with IDHwt gliomas. Our data highlights the importance of considering malignant glioma as a systemic disease that fundamentally alters the immune repertoire in affected patients.

Comparison of Transcriptomic Signatures between Monkeypox-Infected Monkey and Human Cell Lines

Monkeypox virus (MPV) is a smallpox-like virus belonging to the genus Orthopoxvirus of the family Poxviridae. Unlike smallpox with no animal reservoir identified and patients suffering from milder symptoms with less mortality, several animals were confirmed to serve as natural hosts of MPV. The reemergence of a recently reported monkeypox epidemic outbreak in nonendemic countries has raised concerns about a global outburst. Since the underlying mechanism of animal-to-human transmission remains largely unknown, comprehensive analyses to discover principal differences in gene signatures during disease progression have become ever more critical. In this study, two MPV-infected in vitro models, including human immortal epithelial cancer (HeLa) cells and rhesus monkey (Macaca mulatta) kidney epithelial (MK2) cells, were chosen as the two subjects to identify alterations in gene expression profiles, together with co-regulated genes and pathways that are affected during monkeypox disease progression. Using Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG), and MetaCore analyses, we discovered that elevated expression of genes associated with interleukins (ILs), G protein-coupled receptors (GPCRs), heat shock proteins (HSPs), Toll-like receptors (TLRs), and metabolic-related pathways play major roles in disease progression of both monkeypox-infected monkey MK2 and human HeLa cell lines. Interestingly, our analytical results also revealed that a cluster of differentiation 40 (CD40), plasmin, and histamine served as major regulators in the monkeypox-infected monkey MK2 cell line model, while interferons (IFNs), macrophages, and neutrophil-related signaling pathways dominated the monkeypox-infected human HeLa cell line model. Among immune pathways of interest, apart from traditional monkeypox-regulated signaling pathways such as nuclear factor- (NF-κB), mitogen-activated protein kinases (MAPKs), and tumor necrosis factors (TNFs), we also identified highly significantly expressed genes in both monkey and human models that played pivotal roles during the progression of monkeypox infection, including CXCL1, TNFAIP3, BIRC3, IL6, CCL2, ZC3H12A, IL11, CSF2, LIF, PTX3, IER3, EGR1, ADORA2A, and DUOX1, together with several epigenetic regulators, such as histone cluster family gene members, HIST1H3D, HIST1H2BJ, etc. These findings might contribute to specific underlying mechanisms related to the pathophysiology and provide suggestions regarding modes of transmission, post-infectious sequelae, and vaccine development for monkeypox in the future.

Analysis of Glaucoma Associated Genes in Response to Inflammation, an Examination of a Public Data Set Derived from Peripheral Blood from Patients with Hepatitis C.

IntroductionGlaucoma is the second leading cause of blindness worldwide and despite its prevalence, there are still many unanswered questions related to its pathogenesis. There is evidence that oxidative stress and inflammation play a major role in disease progression. Glaucoma patients from several studies showed altered gene expression in leukocytes, revealing the possibility of using peripheral biomarkers to diagnose or stage glaucoma. The fact that glaucoma is associated with gene expression changes in tissues distant from the retina underscores the possible involvement of systemic oxidative stress and inflammation as potential contributing or compounding factors in glaucoma.MethodsWe assembled a list of oxidative stress and inflammatory markers related to glaucoma based on a review of the literature. In addition, we utilized publicly available data sets of gene expression values collected from peripheral blood mononuclear cells and macrophages from two patient groups: those chronically infected by the hepatitis C virus and those who have cleared it. Activation of the innate immune response can render cells or tissues more responsive to a second delayed proinflammatory stimulus. Additional gene expression data from these cells after subsequent polyinosinic:polycytidylic acid treatment, used to elicit an acute inflammatory response, allowed for the investigation of the acute inflammatory response in these groups. We used fold-change comparison values between the two patient groups to identify genes of interest.ResultsA comparison analysis identified 17 glaucoma biomarkers that were differentially expressed in response to HCV-mediated inflammation. Of these 17, six had significant p-values in the baseline vs treated values. Expression data of these genes were compared between patients who had cleared the Hepatitis C virus versus those who had not and identified three genes of interest for further study.DiscussionThese results support our hypothesis that inflammation secondary to Hepatitis C virus infection affects the expression of glaucoma biomarker genes related to the antioxidant response and inflammation. In addition, they provide several potential targets for further research into understanding the relationship between innate responses to viral infection and inflammatory aspects of glaucoma and for potential use as a predictive biomarker or pharmacological intervention in glaucoma.

IMMU-17. SYSTEMIC IMMUNOSUPPRESSION OF CD4+ T HELPER CELLS IN GLIOMA

Abstract Cancer is a systemic disease. Due to the exceedingly rare occurrence of metastasis of cerebral glioma, systemic alterations have, however, not been considered to play a major role in disease progression of glioma. CD4+ T helper (TH) cells orchestrate the adaptive immune response in an antigen-specific, cytokine mediated manner. The aim of our study was to investigate how far cerebral glioma impacts the systemic CD4+ immune repertoire. We therefore analyzed the peripheral blood CD4+ TH cell phenotype and cytokine production in 100 patients with IDHwt, 30 IDHmut and 16 IDHmut 1p19q co-deleted gliomas in comparison with age-matched healthy donors (HD). We found a significant skewing of the peripheral phenotype in IDHwt glioma patients, showing a TH1 expansion and reduced numbers of T follicular helper cells (TFH), TH1* and mucosa associated invariant T (MAIT) cells, while TH2 and TH17 percentages remained stable compared to IDHmut and HD. Interestingly, although TH1 cells were dominant in IDHwt patients, intracellular cytokine staining showed a distinct reduction of IFNg and TNFa production after in vitro stimulation, while IL-4 was significantly increased compared to HD. No alterations between all groups were observed in IL-2, IL-10 or IL-17 production. Profiling of metabolic surface markers further revealed three distinct groups of CD4+ T cells which are altered in IDHwt patients, indicating a metabolic shift in the CD4+ repertoire compared to HD. Taken together, our results show a CD4+ TH cell type specific skewing of the peripheral immune repertoire in patients with IDHwt gliomas. Our data highlights the importance of considering malignant glioma as a systemic disease that fundamentally alters the immune repertoire in affected patients.

P16.03 Cerebral gliom alters the peripheral CD4+ T helper cell phenotype

Abstract BACKGROUND Cancer is a systemic disease. Due to the exceedingly rare occurrence of metastasis of cerebral glioma, systemic alterations have, however, not been considered to play a major role in disease progression of glioma. CD4+ T helper (TH) cells orchestrate the adaptive immune response in an antigen-specific, cytokine mediated manner. The aim of our study was to investigate how far cerebral glioma impacts the systemic CD4+ immune repertoire. MATERIAL AND METHODS We performed flow-cytometry analysis of the peripheral blood CD4+ TH cell phenotype and cytokine production in 100 patients with IDHwt, 30 IDHmut and 16 IDHmut 1p19q co-deleted gliomas in comparison with age-matched healthy donors (HD). Data was analyzed using a Fortessa LSR and Diva software. Multiparameter analyses were performed using UMAP and SpadeVizR trees. The study was approved by the ethics committee (PV4904). RESULTS We found a significant skewing of the peripheral immunophenotype in IDHwt glioma patients, showing a TH1 expansion and reduced numbers of T follicular helper cells (TFH), TH1* and mucosa associated invariant T (MAIT) cells (p<0.001), while TH2 and TH17 percentages remained stable compared to IDHmut and HD. Although TH1 cells were dominant in IDHwt patients (p<0.01), intracellular cytokine staining showed a reduction of IFNγ and TNFα production after in vitro stimulation, while IL-4 was significantly increased compared to HD (p<0.05). No alterations between all groups were observed in IL-2, IL-10 or IL-17 production. Profiling of metabolic surface markers further revealed increased expression of GLUT1 on CD4+ T cells in IDHwt patients, indicating an activated CD4+ repertoire compared to HD. CONCLUSION Taken together, our results show a CD4+ TH cell type specific skewing of the peripheral immune repertoire in patients with IDHwt gliomas. Our data highlights the importance of considering malignant glioma as a disease with profound systemic effects fundamentally altering the immune repertoire in affected patients.

Changing delta hepatitis patient profile: A single center experience in Valencia region, Spain.

BACKGROUNDDelta hepatitis is a rare infection with an aggressive disease course. For almost three decades, however, there have been no epidemiological studies in our traditionally endemic area.AIMTo investigate the prevalence of delta hepatitis in a sample of patients with chronic hepatitis B virus (HBV) infection followed at a Hepatology Unit in Valencia, Spain.METHODSRetrospective evaluation of anti-hepatitis D virus-immunoglobulin G seroprevalence among patients with chronic HBV infection (n = 605) followed at a reference Hepatology Unit in Spain.RESULTSThe prevalence of anti-hepatitis D virus-immunoglobulin G among HBV-infected patients was 11.5%: Male (63%) and median age of 52 years. The majority were born in Spain (67%) and primarily infected through intravenous drug use. However, a significant percent (24.5%), particularly those diagnosed in more recent years, were migrants presumably nosocomially infected. Comorbidities such as diabetes (8.5%), obesity/overweight (55%), and alcohol consumption (34%) were frequent. A high proportion of patients developed liver complications such as cirrhosis (77%), liver decompensation (81%), hepatocellular carcinoma (HCC) (16.5%), or required liver transplantation (LT) (59.5%). Diabetes was associated with progression to cirrhosis, LT, and death. Male sex, increasing age, and alcohol were associated with LT and HCC. Compared to HBV mono-infected patients, delta individuals developed cirrhosis and liver decompensation more frequently, with no differences in HCC rates.CONCLUSIONPatients infected in the 1980’s were mostly locals infected through intravenous drug use, whereas those diagnosed recently are frequently non-Spanish natives from endemic areas. Regardless of their origin, patients are predominantly male with significant comorbidities, which potentially play a major role in disease progression. We confirm a high rate of subsequent liver complications.

Computer-Aided Structure Prediction of Bluetongue Virus Coat Protein VP2 Assisted by Optimized Potential for Liquid Simulations (OPLS).

The capsid coated protein of Bluetongue virus (BTV) VP2 is responsible for BTV transmission by the Culicoides vector to vertebrate hosts. Besides, VP2 is responsible for BTV entry into permissive cells and hence plays a major role in disease progression. However, its mechanism of action is still unknown. The present investigation aimed to predict the 3D structure of Viral Protein 2 of the bluetongue virus assisted by Optimized Potential for Liquid Simulations (OPLS), structure validation, and an active site prediction. The 3D structure of the VP2 protein was built using a Python-based Computational algorithm. The templates were identified using Smith waterman's Local alignment. The VP2 protein structure validated using PROCHECK. Molecular Dynamics Simulation (MDS) studies were performed using an academic software Desmond, Schrodinger dynamics, for determining the stability of a model protein. The Ligand-Binding site was predicted by structure comparison using homology search and proteinprotein network analysis to reveal their stability and inhibition mechanism, followed by the active site identification. The secondary structure of the VP2 reveals that the protein contains 220 alpha helix atoms, 40 310 helix, 151 beta sheets, 134 coils and 424 turns, whereas the 3D structure of Viral Protein 2 of BTV has been found to have 15774 total atoms in the structure. However, 961 amino acids were found in the final model. The dynamical cross-correlation matrix (DCCM) analysis tool identifies putative protein domains and also confirms the stability of the predicted model and their dynamical behavior difference with the correlative fluctuations in motion. The biological interpretation of the Viral Protein 2 was carried out. DCCM maps were calculated, using a different coordinate reference frame, through which, protein domain boundaries and protein domain residue constituents were identified. The obtained model shows good reliability. Moreover, we anticipated that this research should play a promising role in the identification of novel candidates with the target protein to inhibit their functional significance.

Over-expression of Chitotriosidase Modulates ECM Distribution in Atherosclerotic Plaques of Mice

Abstract Macrophage is the predominant cell type in all phases of atherosclerosis and it plays a major role in disease progression. Although mammals have no endogenous chitin, macrophages produce chitotriosidase-1 (CHIT1) as part of the innate immune response in various inflammatory conditions including atherosclerosis. We aimed to investigate mechanisms by which CHIT1 may modulate the progression of atherosclerosis using CHIT1-overexpressing, Ldlr−/− mice, and also to determine whether overexpression of CHIT1 by macrophages affects the morphology of atherosclerotic plaques. We developed an atherosclerosis-prone, conditional CHIT1 over-expressing (CHIT1-OE) mouse model which allowed us to study the effects of macrophage CHIT1 over-expression on morphological characteristics as well as extracellular matrix (ECM) formation in vivo. Further, bone marrow-derived macrophages (BMDM) from CHIT1-OE mice and littermate controls were used to determine the cellular mechanisms of non-enzymatic CHIT1 signaling in vitro. Over-expression of CHIT1 in Ldlr−/− mice after 12 weeks of high-fat diet modulated the ECM by altering the accumulation and localization of hyaluronic acid (HA) and collagen in atherosclerotic plaques observed within the aortic sinus. We also found, in vitro, that macrophage invasiveness was significantly enhanced in BMDM from CHIT1-OE mice upon addition of IL-13 as an immunoregulator. Our findings indicate that CHIT1 over-expression modulates ECM properties in the atherosclerotic plaques of Ldlr−/− mice and macrophage behavior in vitro, which may be atheroprotective and promote plaque stability.

Longitudinal Characterization of Cortical Lesion Development and Evolution in Multiple Sclerosis with 7.0-T MRI.

Background Cortical lesions develop early in multiple sclerosis (MS) and play a major role in disease progression. MRI at 7.0 T shows high sensitivity for detection of cortical lesions as well as better spatial resolution and signal-to-noise ratio compared with lower field strengths. Purpose To longitudinally characterize (a) the development and evolution of cortical lesions in multiple sclerosis across the cortical width, sulci, and gyri; (b) their relation with white matter lesion accrual; and (c) the contribution of 7.0-T cortical and white matter lesion load and cortical thickness to neurologic disability. Materials and Methods Twenty participants with relapsing-remitting MS and 13 with secondary progressive MS, along with 10 age-matched healthy controls, were prospectively recruited from 2010 to 2016 to acquire, in two imaging sessions (mean interval, 1.5 years), 7.0-T MRI T2*-weighted gradient-echo images (0.33 × 0.33 × 1.0 mm3) for cortical and white matter lesion segmentation and 3.0-T T1-weighted images for cortical surface reconstruction and cortical thickness estimation. Cortical lesions were sampled through the cortex to quantify cortical lesion distribution. The Expanded Disability Status Scale (EDSS) was used to assess neurologic disability. Nonparametric statistics assessed differences between and within groups in MRI metrics of cortical and white matter lesion burden; regression analysis explored associations of disability with MRI metrics. Results Twenty-five of 31 (81%) participants developed new cortical lesions per year (intracortical, 1.3 ± 1.7 vs leukocortical, 0.7 ± 1.9; P = .04), surpassing white matter lesion accrual (cortical, 2.0 ± 2.8 vs white matter, 0.7 ± 0.6; P = .01). In contrast to white matter lesions, cortical lesion accrual was greater in participants with secondary progressive MS than with relapsing-remitting MS (3.6 lesions/year ± 4.2 vs 1.1 lesions/year ± 0.9, respectively; P = .03) and preferentially localized in sulci. Total cortical lesion volume independently predicted baseline EDSS (β = 1.5, P < .001) and EDSS changes at follow-up (β = 0.5, P = .003). Conclusion Cortical lesions predominantly develop intracortically and within sulci, suggesting an inflammatory cerebrospinal fluid-mediated lesion pathogenesis. Cortical lesion accumulation was prominent at 7.0 T and independently predicted neurologic disability progression. © RSNA, 2019 Online supplemental material is available for this article. See also the editorial by Filippi and Rocca in this issue.

Epithelial-Mesenchymal Transition during Metastasis of HPV-Negative Pharyngeal Squamous Cell Carcinoma.

In epithelial tumors, a shift towards a mesenchymal phenotype has been associated with increased invasiveness and metastasis. It is assumed that this phenomenon plays a major role in disease progression and ultimately prognosis. This study investigated epithelial-mesenchymal transition (EMT) in human papillomavirus- (HPV-) negative pharyngeal squamous cell carcinoma. Tissue was obtained from one hypopharyngeal primary tumor and a regional lymph node metastasis during surgery with curative intention. A cell culture was established from the primary tumor and mesenchymal growth conditions were emulated. Gene expression profiling was performed (Human 8 × 60 K design array, Agilent Technologies) and EMT was assessed by a gene set (MSigDB: M5930, Hallmark_epithelial_mesenchymal_transition), applying gene set expression analysis (GSEA). Immunohistochemical staining and flow cytometry of CD44 and E-cadherin were compared in primary tumor, metastasis, and cell cultures. Primary tumor and metastasis were highly positive for CD44. A loss of E-cadherin occurred in the metastasis. Flow cytometry showed the appearance of a population without E-cadherin in spheroid colonies. In GSEA, the EMT phenotype was enriched in the primary tumor compared to metastasis and cell cultures (FDR < 25%, p < 5%). EMT showed variable expression during metastasis. It may thereby be a dynamic state in HPV-negative pharyngeal squamous cell carcinoma that is active only during the process of metastasis itself. Thereby, the primary tumor as well as the metastasis may exhibit fewer EMT properties.

The physician-scientists: rare species in Africa.

There is paucity of physician-scientists in Africa, resulting in overt dependence of clinical practice on research findings from advanced “first world” countries. Physician-scientists include individuals with a medical degree alone or combined with other advanced degrees (e.g. MD/MBChB and PhD) with a career path in biomedical/ translational and patient-oriented/evaluative science research. The paucity of clinically trained research scientists in Africa could result in dire consequences as exemplified in the recent Ebola virus epidemic in West Africa, where shortage of skilled clinical scientists, played a major role in disease progression and mortality. Here we contextualise the role of physician-scientist in health care management, highlight factors limiting the training of physician-scientist in Africa and proffer implementable recommendations to address these factors.

Crimean-Congo Hemorrhagic Fever: Tick-Host-Virus Interactions.

Crimean-Congo hemorrhagic fever virus (CCHFV) is transmitted to humans by bite of infected ticks or by direct contact with blood or tissues of viremic patients or animals. It causes to humans a severe disease with fatality up to 30%. The current knowledge about the vector-host-CCHFV interactions is very limited due to the high-level containment required for CCHFV studies. Among ticks, Hyalomma spp. are considered the most competent virus vectors. CCHFV evades the tick immune response, and following its replication in the lining of the tick's midgut, it is disseminated by the hemolymph in the salivary glands and reproductive organs. The introduction of salivary gland secretions into the host cells is the major route via which CCHFV enters the host. Following an initial amplification at the site of inoculation, the virus is spread to the target organs. Apoptosis is induced via both intrinsic and extrinsic pathways. Genetic factors and immune status of the host may affect the release of cytokines which play a major role in disease progression and outcome. It is expected that the use of new technology of metabolomics, transcriptomics and proteomics will lead to improved understanding of CCHFV-host interactions and identify potential targets for blocking the CCHFV transmission.

Targeting H19, an Imprinted Long Non-Coding RNA, in Hepatic Functions and Liver Diseases.

H19 is a long non-coding RNA regulated by genomic imprinting through methylation at the locus between H19 and IGF2. H19 is important in normal liver development, controlling proliferation and impacting genes involved in an important network controlling fetal development. H19 also plays a major role in disease progression, particularly in hepatocellular carcinoma. H19 participates in the epigenetic regulation of many processes impacting diseases, such as activating the miR-200 pathway by histone acetylation to inhibit the epithelial-mesenchymal transition to suppress tumor metastasis. Furthermore, H19’s normal regulation is disturbed in diseases, such as hepatocellular carcinoma. In this disease, aberrant epigenetic maintenance results in biallelic expression of IGF2, leading to uncontrolled cellular proliferation. This review aims to further research utilizing H19 for drug discovery and the treatment of liver diseases by focusing on both the epigenetic regulation of H19 and how H19 regulates normal liver functions and diseases, particularly by epigenetic mechanisms.

Proteomic level changes associated with S3I201 treated U87 glioma cells

Glioblastoma multiforme is Grade IV brain tumor associated with high mortality and limited therapeutics. Signal Transducer and Activator of Transcription 3 (STAT3) is persistently active in several cancers including gliomas, and plays a major role in disease progression and survival of glioma patients, thus being a potential therapeutic target for treatment. S3I201 and its analogs inhibit the transcriptional functions of STAT3 and reduce growth of tumor tissues. Here we have studied proteomic alteration associated with S3I201 treated U87 cells using 2-DE and Isobaric tags for relative and absolute quantitation coupled with mass spectrometry. This analysis revealed 136 differentially expressed proteins which were functionally classified with gene ontology analysis. Results showed metabolism, apoptosis, cytoskeletal behaviour, cell redox homeostasis and immune response as the most affected biological processes on S3I201 treatment. Apoptosis-inducing factor 1 mitochondrial, cyclophilin A and chloride intra-cellular channel protein 1 were found to be up-regulated which possibly contributes to its anti-tumorigenic function. Several glycolytic enzymes like phosphoglycerate mutase 1 were also found to be up-regulated and its expression was validated using immunoblot. Conclusively, our study shows the downstream effects of S3I201 in U87 glioma cells and suggests its therapeutic potential. SignificanceGliomas with constitutive expression can be treated with STAT3 inhibitors. S3I201, a STAT3 inhibitor, reduces the growth of glioma cells thus could be studied further for its application as anti-glioma agent. This study investigated proteomic alteration associated with S3I201 in U87 cells using complementary proteomic approaches, and our findings suggest that S3I201 influences central metabolism, apoptosis, cytoskeletal behaviour, cell redox homeostasis and immune response as the most affected biological processes which altogether contribute to its anti-tumorigenic activity. Several proteins were identified which may serve as prognostic or predictive markers in GBM. Apoptosis-inducing factor 1 mitochondrial and cyclophilin A were identified as potential therapeutic targets and further investigations on these candidates may facilitate therapeutic development and suggests that GBM therapy can be improved by targeting cellular metabolism and by using immunotherapy.

Vulnerability of calbindin, calretinin and parvalbumin in a transgenic/knock-in APPswe/PS1dE9 mouse model of Alzheimer disease together with disruption of hippocampal neurogenesis

The pathogenesis of Alzheimer disease (AD) is characterized by accumulation of β-amyloid protein in the brain (in both soluble and insoluble forms) and by the presence of intracellular neurofibrillary tangles (NFTs), leading to neurotoxicity. The exact mechanisms whereby Aβ triggers brain alterations are unclear. However, accumulating evidence suggests that a deregulation of Ca2+ signaling may play a major role in disease progression. Calcium-buffering proteins, including calbindin-D28K (CB), calretinin (CR) and parvalbumin (PV), may offer neuroprotection by maintaining calcium homeostasis. Although marked reductions in these proteins have been observed in the brains of mice and humans with AD, their contribution to AD pathology remains unclear. The aim of the present study was to analyze distribution patterns of CB+, CR+ and PV+ interneurons in different areas of the hippocampus, a brain region that is severely affected in AD. A transgenic knock-in APPswe/PS1dE9 mouse model of familial AD was used. The data were obtained from the brains of 3- and 12-month-old animals. These ages roughly correspond to an early mature adult (prior to clinical manifestations) and a late middle-age (clinical symptoms readily detectable) phase in human AD patients. Immunostaining revealed increases in CB and PV immunoreactivity (IR) in the hippocampus of 3-month-old transgenic mice, compared to wild-type animals. Possibly, these proteins are upregulated in an attempt to control cellular homeostasis and synaptic plasticity. However, the pattern of CB-IR was reversed in 12-month-old animals, potentially indicating a loss of cellular capacity to respond to pathophysiological processes. In addition, at this age, a noticeable increase in PV-IR was observed, suggesting the presence of hippocampal network hyperactivity in older AD-like mice. Our results indicate that CaBP+ neuronal subpopulations play a role in adult neurogenesis and in AD pathology, particularly at early disease stages, suggesting that these neurons may serve as potential predictors of future AD in non-demented individuals.

A novel anti-inflammatory natural product from Sphaeranthus indicus inhibits expression of VCAM1 and ICAM1, and slows atherosclerosis progression independent of lipid changes.

A large body of evidence suggests that atherosclerosis is an inflammatory disease, in which cytokines and growth factors play a major role in disease progression. The methanolic extracts of Sphaeranthus indicus as well as its active ingredient, 7-hydroxy frullanoide (7-HF), are shown to suppress LPS-induced cytokine production from mononuclear cells, and inhibit the expression of VCAM1, ICAM1 and E-selectin by TNF-α- stimulated HUVECs in a concentration-dependent manner. We tested the hypothesis that the inhibition of cytokines and adhesion molecules should attenuate the progression of atherosclerosis, independent of changes in the lipid profile. Studies were carried out in two animal models: a high fat-fed LDLr-/- mouse and a high fat-fed hyperlipidemic hamster. Methanolic extract of S. indicus was dosed to hyperlipidemic LDLr-/- at 100 and 300 mg (equivalent to 20 and 60 mg 7-HF)/kg body weight/ day for 8 weeks, and plasma lipids as well as aortic lesion area were quantitated. Hyperlipidemic hamsters were treated with one dose of 200 mg/kg/day. S. indicus extract treatment did not alter the lipid profile in both animal models, but reduced aortic lesion area in LDLr-/- mice and hyperlipidemic hamsters by 22 % and 45 %, respectively. Fenofibrate, included as a reference agent, decreased aortic lesions by 26 % in LDLr -/- mice and 84 % in hyperlipidemic hamsters, respectively, which was driven by massive reductions in proatherogenic lipoproteins. The lipid-independent anti-atherosclerotic activity of S. indicus was associated with the reductions in the circulating levels of MCP-1, TNF-α, and IL-6 via phosphorylation and degradation of IkB-α that prevents translocation of NF-kB in the nucleus to induce proinflammatory cytokines. Our findings demonstrate that anti-inflammatory agents that lower pro-inflammatory proteins inhibit the progression of atherosclerosis. The methanolic extract of S. inducus, currently being used to treat psoriasis, offer promise to benefit individuals who have high circulating pro-inflammatory cytokines, and predisposed to coronary artery disease.

Astrocyte response to IFN-γ limits IL-6-mediated microglia activation and progressive autoimmune encephalomyelitis.

BackgroundTherapeutic modalities effective in patients with progressive forms of multiple sclerosis (MS) are limited. In a murine model of progressive MS, the sustained disability during the chronic phase of experimental autoimmune encephalomyelitis (EAE) correlated with elevated expression of interleukin (IL)-6, a cytokine with pleiotropic functions and therapeutic target for non-central nervous system (CNS) autoimmune disease. Sustained IL-6 expression in astrocytes restricted to areas of demyelination suggested that IL-6 plays a major role in disease progression during chronic EAE.MethodsA progressive form of EAE was induced using transgenic mice expressing a dominant negative interferon-γ (IFN-γ) receptor alpha chain under control of human glial fibrillary acidic protein (GFAP) promoter (GFAPγR1Δ mice). The role of IL-6 in regulating progressive CNS autoimmunity was assessed by treating GFAPγR1Δ mice with anti-IL-6 neutralizing antibody during chronic EAE.ResultsIL-6 neutralization restricted disease progression and decreased disability, myelin loss, and axonal damage without affecting astrogliosis. IL-6 blockade reduced CNS inflammation by limiting inflammatory cell proliferation; however, the relative frequencies of CNS leukocyte infiltrates, including the Th1, Th17, and Treg CD4 T cell subsets, were not altered. IL-6 blockade rather limited the activation and proliferation of microglia, which correlated with higher expression of Galectin-1, a regulator of microglia activation expressed by astrocytes.ConclusionsThese data demonstrate that astrocyte-derived IL-6 is a key mediator of progressive disease and support IL-6 blockade as a viable intervention strategy to combat progressive MS.