Dp71 expression in human glioblastoma

Background: Dp71 is the most abundant dystrophin (DMD) gene product in the nervous system. Mutation in the Dp71 coding region is associated with cognitive disturbances in Duchenne muscular dystrophy (DMD) patients, but the function of dystrophin Dp71 in tumor progression remains to be established. This study investigated Dp71 expression in glioblastoma, the most common and aggressive primary tumor of the central nervous system (CNS). Methods: Dp71 expression was analyzed by immunofluorescence, immunohistochemistry, RT-PCR, and immunoblotting in glioblastoma cell lines and cells isolated from human glioblastoma multiforme (GBM) bioptic specimens. Results: Dp71 isoform was expressed in normal human astrocytes (NHA) cell lines and decreased in glioblastoma cell lines and cells isolated from human glioblastoma multiforme bioptic specimens. Moreover, Dp71 was localized in the nucleus in normal cells, while it was localized into the cytoplasm of glioblastoma cells organized in clusters. We have shown, by double labeling, that Dp71 colocalizes with lamin B in normal astrocytes cells, confirming the roles of Dp71 and lamin B in maintaining nuclear architecture. Finally, we demonstrated that decreased Dp71 protein in cells isolated from human bioptic specimens was inversely correlated with the Ki-67 tumor proliferative index. Conclusion: A decreased Dp71 expression is associated with cancer proliferation and poor prognosis in glioblastoma

Primary immunodeficiency

Primary immunodeficiency disorder (PID) refers to a heterogeneous group of over 130 disorders that result from defects in immune system development and/or function. PIDs are broadly classified as disorders of adaptive immunity (i.e., T-cell, B-cell or combined immunodeficiencies) or of innate immunity (e.g., phagocyte and complement disorders). Although the clinical manifestations of PIDs are highly variable, most disorders involve at least an increased susceptibility to infection. Early diagnosis and treatment are imperative for preventing significant disease-associated morbidity and, therefore, consultation with a clinical immunologist is essential. PIDs should be suspected in patients with: recurrent sinus or ear infections or pneumonias within a 1 year period; failure to thrive; poor response to prolonged use of antibiotics; persistent thrush or skin abscesses; or a family history of PID. Patients with multiple autoimmune diseases should also be evaluated. Diagnostic testing often involves lymphocyte proliferation assays, flow cytometry, measurement of serum immunoglobulin (Ig) levels, assessment of serum specific antibody titers in response to vaccine antigens, neutrophil function assays, stimulation assays for cytokine responses, and complement studies. The treatment of PIDs is complex and generally requires both supportive and definitive strategies. Ig replacement therapy is the mainstay of therapy for B-cell disorders, and is also an important supportive treatment for many patients with combined immunodeficiency disorders. The heterogeneous group of disorders involving the T-cell arm of the adaptive system, such as severe combined immunodeficiency (SCID), require immune reconstitution as soon as possible. The treatment of innate immunodeficiency disorders varies depending on the type of defect, but may involve antifungal and antibiotic prophylaxis, cytokine replacement, vaccinations and bone marrow transplantation. This article provides a detailed overview of the major categories of PIDs and strategies for the appropriate diagnosis and management of these rare disorders

Missense mutations in the Fas gene resulting in autoimmune lymphoproliferative syndrome: a molecular and immunological analysis

Programmed cell death (or apoptosis) is a physiological process essential to the normal development and homeostatic maintenance of the immune system. The Fas/Apo-1 receptor plays a crucial role in the regulation of apoptosis, as demonstrated by lymphoproliferation in MRL-lpr/lpr mice and by the recently described autoimmune lymphoproliferative syndrome (ALPS) in humans, both of which are due to mutations in the Fas gene. We describe a novel family with ALPS in which three affected siblings carry two distinct missense mutations on both the Fas gene alleles and show lack of Fas-induced apoptosis. The children share common clinical features including splenomegaly and lymphadenopathy, but only one developed severe autoimmune manifestations. In all three siblings, we demonstrated the presence of anergic CD3+CD4-CD8- (double negative, [DN]) T cells; moreover, a chronic lymphocyte activation was found, as demonstrated by the presence of high levels of HLA-DR expression on peripheral CD3+ cells and by the presence of high levels of serum activation markers such as soluble interleukin-2 receptor (slL-2R) and soluble CD30 (sCD30)

Pro‑differentiating compounds for human intervertebral disc cells are present in Violina pumpkin leaf extracts

The intervertebral disc degeneration (IDD) is closely associated with inflammation, oxidative stress and loss of the discogenic phenotype which current therapies are unable to reverse. Here, the effects of acetone extract from Violina pumpkin (Cucurbita moschata) leaves on degenerated intervertebral disc (IVD) cells was investigated. IVD cells were isolated from degenerated disc tissue of patients undergoing spinal surgery, and exposed to acetone extract and three major thin layer chromatography subfractions. We found that the cells benefit from exposure in particular to subfraction 7 consisting almost entirely of p-Coumaric acid. Subfraction 7-treated cells showed a significant increase of discogenic transcription factors (SOX9, TRPS1), extracellular matrix components (aggrecan, collagen type II), cellular homeostasis and stress response regulators (FOXO3a, Nrf2, SOD2, SIRT1). Migratory ability and the expression of OCT4, two important markers related to the presence and activity of stem cells also increased. Moreover, subfraction 7 counteractes H2O2-triggered cell damage preventing in particular the increase of the pro-inflammatory and antichondrogenic microRNA, miR-221. This strengthens the hypothesis that adequate stimuli can support resident cells to repopulate the degenerate IVD and restart the anabolic machinery. Taken together, the data we obtained contribute to the discovery of molecules potentially effective in slowing the progression of IDD, a disease for which there is currently no effective treatment. Moreover, the enhancement of a part of plant, the pumpkin leaves, considered a waste product in the Western world, demonstrating that it contains substances with potential beneficial effects on human health

A mutation in caspase-9 decreases the expression of BAFFR and ICOS in patients with immunodeficiency and lymphoproliferation

Lymphocyte apoptosis is mainly induced by either death receptor-dependent activation of caspase-8 or mitochondria-dependent activation of caspase-9. Mutations in caspase-8 lead to autoimmunity/lymphoproliferation and immunodeficiency. This work describes a heterozygous H237P mutation in caspase-9 that can lead to similar disorders. H237P mutation was detected in two patients: Pt1 with autoimmunity/lymphoproliferation, severe hypogammaglobulinemia and Pt2 with mild hypogammaglobulinemia and Burkitt lymphoma. Their lymphocytes displayed defective caspase-9 activity and decreased apoptotic and activation responses. Transfection experiments showed that mutant caspase-9 display defective enzyme and proapoptotic activities and a dominant-negative effect on wild-type caspase-9. Ex vivo analysis of the patients' lymphocytes and in vitro transfection experiments showed that the expression of mutant caspase-9 correlated with a downregulation of BAFFR (B-cell-activating factor belonging to the TNF family (BAFF) receptor) in B cells and ICOS (inducible T-cell costimulator) in T cells. Both patients carried a second inherited heterozygous mutation missing in the relatives carrying H237P: Pt1 in the transmembrane activator and CAML interactor (TACI) gene (S144X) and Pt2 in the perforin (PRF1) gene (N252S). Both mutations have been previously associated with immunodeficiencies in homozygosis or compound heterozygosis. Taken together, these data suggest that caspase-9 mutations may predispose to immunodeficiency by cooperating with other genetic factors, possibly by downregulating the expression of BAFFR and ICO

The effects of iCVD film thickness and conformality on the permeability and wetting of MD membranes

Membranes possessing high permeability to water vapor and high liquid entry pressure (LEP) are necessary for efficient membrane distillation (MD) desalination. A common technique to prepare specialized MD membranes consists of coating a hydrophilic or hydrophobic base membrane with a low surface-energy material. This increases its liquid entry pressure, making the membrane suitable for MD. However, in addition to increasing LEP, the surface-coating may also decrease permeability of the membrane by reducing its average pore size. In this study, we quantify the effects of initiated chemical vapor deposition (iCVD) polymer coatings on membrane permeability and LEP. We consider whether the iCVD films should have minimized thickness or maximized non-conformality, in order to maximize the permeability achieved for a given value of LEP. We determined theoretically that permeability of a single pore is maximized with a highly non-conformal iCVD coating. However, the overall permeability of a membrane consisting of many pores is maximized when iCVD film thickness is minimized. We applied the findings experimentally, preparing an iCVD-treated track-etched polycarbonate (PCTE) membrane and testing it in a permeate gap membrane distillation (PCMD) system. This study focuses on membranes with clearly defined, cylindrical pores. However, we believe that the principles we discuss will extend to membranes with more complex pore architectures. Overall, this work indicates that the focus of surface-coating development should be on minimizing film thickness, not on increasing their non-conformality.MIT & Masdar Institute Cooperative Program (02/MI/MI/CP/11/07633/GEN/G/00)Massachusetts Institute of Technology. Institute for Soldier Nanotechnologies (W911NF-13-d-0001

Mutation@A Glance: An Integrative Web Application for Analysing Mutations from Human Genetic Diseases

Although mutation analysis serves as a key part in making a definitive diagnosis about a genetic disease, it still remains a time-consuming step to interpret their biological implications through integration of various lines of archived information about genes in question. To expedite this evaluation step of disease-causing genetic variations, here we developed Mutation@A Glance (http://rapid.rcai.riken.jp/mutation/), a highly integrated web-based analysis tool for analysing human disease mutations; it implements a user-friendly graphical interface to visualize about 40 000 known disease-associated mutations and genetic polymorphisms from more than 2600 protein-coding human disease-causing genes. Mutation@A Glance locates already known genetic variation data individually on the nucleotide and the amino acid sequences and makes it possible to cross-reference them with tertiary and/or quaternary protein structures and various functional features associated with specific amino acid residues in the proteins. We showed that the disease-associated missense mutations had a stronger tendency to reside in positions relevant to the structure/function of proteins than neutral genetic variations. From a practical viewpoint, Mutation@A Glance could certainly function as a ‘one-stop’ analysis platform for newly determined DNA sequences, which enables us to readily identify and evaluate new genetic variations by integrating multiple lines of information about the disease-causing candidate genes

Clinical heterogeneity and diagnostic delay of autoimmune polyendocrinopathy-candidiasis-ectodermal dystrophy syndrome

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