Unusual clinical presentation of cervical extradural meningioma detected with neuromuscular ultrasound: A case report.

Extradural meningiomas are rare in the cervical region. A total of 70-77% of reported cases have occurred in the thoracic region. Tumors that occur in the cervical region may invade the adjacent nerve root and brachial plexus. Typically, diagnoses of extradural meningioma are made after patients present with signs of myelopathy, such as progressive paresis and numbness. In the current study, a 64-year-old male patient presented with neck pain, numbness and mild weakness in the left hand over a 6-month period. The general neurological examination was unremarkable, except for mild grasping weakness on the left side. Needle electromyography revealed complex repetitive discharges in the left 5 and 6th cervical paraspinal muscles. Neuromuscular ultrasound revealed a lesion over the left 7th cervical root, which enabled the early detection of an extradural meningioma before notable focal neurological defects developed. The patient underwent a subtotal tumor excision, followed by radiotherapy for residual tumor. Histopathological examination confirmed atypical meningioma.

Diagnosis and Clinical Features in Autoimmune-Mediated Movement Disorders.

Movement disorders are common manifestations in autoimmune-mediated encephalitis. This group of diseases is suspected to be triggered by infection or neoplasm. Certain phenotypes correlate with specific autoantibody-related neurological disorders, such as orofacial-lingual dyskinesia with N-methyl-D-aspartate receptor encephalitis and faciobrachial dystonic seizures with leucine-rich glioma-inactivated protein 1 encephalitis. Early diagnosis and treatment, especially for autoantibodies targeting neuronal surface antigens, can improve prognosis. In contrast, the presence of autoantibodies against intracellular neuronal agents warrants screening for underlying malignancy. However, early clinical diagnosis is challenging because these diseases can be misdiagnosed. In this article, we review the distinctive clinical phenotypes, magnetic resonance imaging findings, and current treatment options for autoimmune-mediated encephalitis.

Peripheral host T cells survive hematopoietic stem cell transplantation and promote graft-versus-host disease.

Graft-versus-host disease (GVHD) is a major cause of morbidity and mortality in hematopoietic stem cell transplantation (HSCT). Donor T cells are key mediators in pathogenesis, but a contribution from host T cells has not been explored, as conditioning regimens are believed to deplete host T cells. To evaluate a potential role for host T cells in GVHD, the origin of skin and blood T cells was assessed prospectively in patients after HSCT in the absence of GVHD. While blood contained primarily donor-derived T cells, most T cells in the skin were host derived. We next examined patient skin, colon, and blood during acute GVHD. Host T cells were present in all skin and colon acute GVHD specimens studied, yet were largely absent in blood. We observed acute skin GVHD in the presence of 100% host T cells. Analysis demonstrated that a subset of host T cells in peripheral tissues were proliferating (Ki67+) and producing the proinflammatory cytokines IFN-γ and IL-17 in situ. Comparatively, the majority of antigen-presenting cells (APCs) in tissue in acute GVHD were donor derived, and donor-derived APCs were observed directly adjacent to host T cells. A humanized mouse model demonstrated that host skin-resident T cells could be activated by donor monocytes to generate a GVHD-like dermatitis. Thus, host tissue-resident T cells may play a previously unappreciated pathogenic role in acute GVHD.

CHD1L Regulated PARP1-Driven Pluripotency and Chromatin Remodeling During the Early-Stage Cell Reprogramming.

PARP1 and poly(ADP-ribosyl)ation (PARylation) have been shown to be essential for the initial steps of cellular reprogramming. However, the mechanism underlying PARP1/PARylation-regulated activation of pluripotency loci remains undetermined. Here, we demonstrate that CHD1L, a DNA helicase, possesses chromatin remodeling activity and interacts with PARP1/PARylation in regulating pluripotency during reprogramming. We found that this interaction is mediated through the interplay of the CHD1L macro-domain and the PAR moiety of PARylated-PARP1. Chromatin immunoprecipitation assays demonstrated the co-occupancy of CHD1L and PARP1 at Pou5f1, Nanog, and Esrrb pluripotency loci. Knockdown of CHD1L significantly blocked the binding activity of PARP1 at pluripotency loci and inhibited the efficiency of PARP1-driven reprogramming. Notably, we found that CHD1L-promoted reprogramming requires both a PARP1-interacting domain and DNA helicase activity, partly contributing to the chromatin-remodeling states of pluripotency loci. Taken together, these results identify CHD1L as a key chromatin remodeler involved in PARP1/PARylation-regulated early-stage reprogramming and pluripotency in stem cells.

Biological evaluation of secondary metabolites from the roots of Myrica adenophora.

Bioassay-guided fractionation of the roots of Myrica adenophora led to isolation of 24 known compounds and hitherto unknown compounds, including three A-type proanthocyanidins [adenodimerins A-C], two esters of sucrose [myricadenins A and B ], and the phenolic glycoside 6'-O-galloyl orbicularin. Spectroscopic analyses were used to determine their structures. Adenodimerin A, myricananin C, and myricetin showed strong 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities, with SC50 values of 7.9, 16.3, and 15.9 µM, respectively. Adenodimerin A, myricanone, myricananin C, (-)-myricanol, myricanol 11-O-ß-D-glucopyranoside, and myricetin showed stronger 2,2'-azino-bis-3-ethylbenzthiazoline-6-sulphonic acid (ABTS) radical scavenging activities than the positive control, with SC50 values of 7.5, 19.6, 12.0, 22.3, 19.6, and 15.6 µM, respectively. 5-Deoxymyricanone, porson, 12-hydroxymyricanone (-)-myricanol, and (+)-galeon exhibited anti-tubercular activity against Mycobacterium tuberculosis H37Rv in vitro and MICs values of 25.8, 40.0, 35.8, 30.0, and 15.0 µg/mL, respectively. Myricadenin A, myricanone, myricananin C, and (-)-myricanol exhibited anti-inflammatory activities in the iNOS assay with EC50 values of 18.1, 1.00, 13.0, and 7.5 µM, respectively.

Melamine induces human renal proximal tubular cell injury via transforming growth factor-ß and oxidative stress.

Although several reports have failed to observe adverse subchronic renal effects following relatively high melamine exposure, the safety of low and continuous melamine exposure is still debatable. Recent studies suggest that long-term, low-dose melamine exposure is associated with an increased risk of urolithiasis, which has been linked to chronic kidney disease (CKD). CKD is a consequence of nephron loss and is associated with the interaction of inflammation, oxidative stress, and transforming growth factor-ß (TGF-ß), which increases extracellular matrix genes and cell apoptosis with progression to fibrosis and end-stage renal disease. Thus far, information is still lacking regarding the influence of melamine at the gene and protein levels, which are activated at a much earlier phase than the occurrence of the renal morphological change. In this study, we stimulated human renal proximal tubular HK-2 cells with melamine (0, 125, 250, 500, or 1000 µg/ml) for different time intervals and observed its effects on several well-documented molecular mechanisms of CKD. Here, we demonstrate that melamine can activate mitogen-activated protein kinases, NFκB, and reactive oxygen species, which results in the upregulation of interleukin-6, monocyte chemoattractant protein-1, vascular cell adhesion molecule-1, and TGF-ß1 in HK-2 cells. The melamine-stimulated overexpression of TGF-ß1 not only promotes fibronectin production but also leads to decreased antiapoptotic (bcl-2, bcl-xl)/proapoptotic (bad, bax) protein ratio, increased caspase-3 and caspase-9 activities, and eventually HK-2 cell apoptosis. Our study suggests that melamine exposure may be a risk factor for the chronic loss of tubular cells and may ultimately lead to tubulointerstitial damage.

DDA3 stabilizes microtubules and suppresses neurite formation.

We have previously shown that DDA3 - also known as proline/serine-rich coiled-coil protein 1 (PSRC1) - is a microtubule-associated protein that promotes cell growth by stimulating the ß-catenin pathway. Here, we report that DDA3 can bundle and stabilize microtubules in vivo and in vitro. We found that overexpression of DDA3 increased the abundance of acetylated and tyrosinated microtubules. We employed PC12 and N2a cell lines, as well as cultured hippocampal neurons, and demonstrated that overexpression of DDA3 suppressed neurite/axon outgrowth, whereas its depletion accelerated neurite/axon formation and elongation. Knockdown of DDA3 reduced ß3-tubulin levels in N2a cells, which contributed to the spontaneous neurite formation caused by DDA3 depletion. Consistent with its role in suppressing neuritogenesis, DDA3 was downregulated during induced neuronal differentiation. Moreover, expression of DDA3 was detected in the rat brain at embryonic (E) day E15 and in the cortical region at E17, the period of active neurogenesis. Levels of cortical DDA3 decreased at the beginning of E19, when active neuritogenesis is completed. Overall our results demonstrate that DDA3 is a so-far-unknown microtubule-stabilizing protein that is involved in regulating neurite formation and elongation.

Targeting autophagy enhances BO-1051-induced apoptosis in human malignant glioma cells.

PURPOSE: BO-1051 is an N-mustard derivative that is conjugated with DNA-affinic 9-anilinoacridine. Since BO-1051 was reported to have strong anticancer activity, we investigated the effect and underlying mechanism of BO-1051 in human glioma cell lines. METHODS: Human glioma cell lines U251MG and U87MG were studied with BO-1051 or the combination of BO-1051 and autophagic inhibitors. Growth inhibition was assessed by MTT assay. Apoptosis was measured by annexin V staining followed by flow cytometry and immunoblotting for apoptosis-related molecules. Induction of autophagy was detected by acridine orange labeling, electron microscopy, LC3 localization and its conversion. Transfection of shRNA was used to determine the involvement of Beclin1 in apoptotic cell death. RESULTS: MTT assay showed that BO-1051 suppressed the viability of four glioma cell lines (U251MG, U87MG, GBM-3 and DBTRG-05MG) in a dose-dependent manner. The IC(50) values of BO-1051 for the glioma cells were significantly lower than the values for primary neurons cultures and normal fibroblast cells. Moreover, BO-1051 not only induced apoptotic cell death, but also enhanced autophagic flux via inhibition of Akt/mTOR and activation of Erk1/2. Importantly, suppression of autophagy by 3-methyladenine or bafilomycin A1 significantly increased BO-1051-induced apoptotic cell death in U251MG and U87MG cells. In addition, the proportion of apoptotic cells after BO-1051 treatment was enhanced by co-treatment with shRNA against Beclin1. CONCLUSIONS: BO-1051 induced both apoptosis and autophagy, and inhibition of autophagy significantly augmented the cytotoxic effect of BO-1051. Thus, a combination of BO-1051 and autophagic inhibitors offers a potentially new therapeutic modality for the treatment of malignant glioma.

Betel nut extract and arecoline block insulin signaling and lipid storage in 3T3-L1 adipocytes.

According to several population-based studies, betel nut chewing is associated with metabolic syndrome and diabetes in British South Asians and Taiwanese. However, the underlying molecular mechanism is not yet clear. Arecoline is an alkaloid-type natural product found in betel nuts. Our aim was to clarify the influence of betel nut extract and arecoline on lipid accumulation and insulin signaling in adipocytes. We found that betel nut extract and arecoline blocked lipid storage in 3T3-L1 adipocytes. The possible mechanism may function by inhibiting the expression of the insulin receptor, glucose transporter-4, fatty acid synthase, and the lipid droplet proteins perilipin and adipophilin. In addition, betel nut extract and arecoline increased the basal level of IRS-1 serine(307) phosphorylation and decreased insulin-stimulated IRS-1 tyrosine, Akt, and PI3 kinase phosphorylation. In conclusion, betel nut extract and arecoline have diabetogenic potential on adipocytes that may result in insulin resistance and diabetes at least in part via the obstruction of insulin signaling and the blockage of lipid storage.

Rad is a p53 direct transcriptional target that inhibits cell migration and is frequently silenced in lung carcinoma cells.

The p53 tumor suppressor exerts its function mainly as a transcriptional activator. Here we show that the Ras-related small GTPase Rad, an inhibitor of Rho kinase, is a direct transcriptional target of p53. Expression of Rad messenger RNA (mRNA) and protein was induced by DNA damage in a p53-dependent manner. The -2934/-2905-bp Rad promoter region, to which p53 bound, was required for p53-mediated Rad gene activation. Treatment by DNA damaging agents increased p53 occupancy and histone acetylation in the region of Rad promoter containing the p53-binding site. Expression of Rad diminished the inhibitory phosphorylation at Ser3 of cofilin, a regulator of actin dynamics, and suppressed migration and invasiveness of cancer cells. Knockdown of Rad promoted cell migration and alleviated the p53-mediated migration suppression. Frequent loss of Rad mRNA and protein expression was observed in non-small cell lung carcinoma tissues. Together our results reveal a mechanism that p53 may inhibit cell migration by disrupting actin dynamics via Rad activation and implicate a tumor suppressor role of Rad in lung cancer.

p53 target DDA3 binds ASPP2 and inhibits its stimulation on p53-mediated BAX activation.

The p53 tumor suppressor functions in maintaining the integrity of the genome. We have previously reported that DDA3 is an oncoprotein transcriptionally regulated by p53. To explore mechanisms underlying DDA3 action, we searched for its interacting proteins by yeast two-hybrid screening, and identified ASPP2, a p53 binding protein, as its binding partner. The DDA3/ASPP2 binding was confirmed in vitro by GST pull-down and in vivo by immunofluorescence assay, which indicated colocalization of DDA3 and ASPP2. Interacting domain of DDA3 was mapped to amino acids 118-241, whereas both the N- and C-terminal regions of ASPP2 were capable of binding to DDA3. DDA3 dose-dependently inhibited ASPP2 in stimulating the p53-mediated BAX promoter activation without interfering the binding of ASPP2 to p53. Together these results identify ASPP2 as a bona fide DDA3 interacting protein, and suggest that the ASPP2/DDA3 interaction may inhibit ASPP2 in stimulating the apoptotic signaling of p53.

A case-control study of the HER2 Ile655Val polymorphism and risk of breast cancer in Taiwan.

OBJECTIVES: To investigate the HER2 Ile655Val polymorphism in relation to risk of breast cancer in a case-control study in Taiwan. DESIGN AND METHODS: The HER2 polymorphism at codon 655 was analyzed in 424 patients with breast cancer and 318 controls by using the polymerase chain reaction methodology, followed by the restriction fragment-length polymorphism (PCR-RFLP) analysis. RESULTS: There was a 1.48-fold (95% CI=1.00-2.24) increase in the risk of patients with breast cancer who are Val carrier (Ile/Val and Val/Val genotypes). Furthermore, for the early onset (less than 45 years old) breast cancers with Val carrier, there was a 2.24-fold (95% CI=1.17-4.34) increase in the risk of breast cancer. CONCLUSIONS: Our results indicate that the Val carrier was associated with increased risks in patients with breast cancer in Taiwan. The association was more apparent in patients who were younger than or equal to 45 years of age.

Secreted expression of the classical swine fever virus glycoprotein E(rns) in yeast and application to a sandwich blocking ELISA.

E(rns) is an envelope glycoprotein of classical swine fever virus (CSFV) with RNase activity. The purpose of this study was to produce an active E(rns) for further applications using the yeast secreted expression system. The E(rns) gene was cloned into the expression vector pGAPZalphaC which was introduced into Pichia pastoris. Expression of E(rns) protein in culture supernatant was confirmed by Western blot analysis using both the monoclonal antibody against CSFV E(rns) and CSFV-positive swine serum. The yeast-expressed E(rns) (yE(rns)) was shown to have N-linked glycosylation and to form homodimer of 74 kDa molecules. All monomer, homodimer, and deglycosylated forms of yE(rns) demonstrated intrinsic ribonuclease activity and a clear preference for uridine-rich sequence. A direct sandwich blocking enzyme-linked immunosorbent assay (ELISA) based on the yE(rns) was developed with a high sensitivity and specificity. The yE(rns) which possesses enzymatic activity and retains antigenicity may provide a useful material for developing a diagnostic kit.

Cloning and characterization of an iron regulated locus, iroA, in Salmonella enterica serovar Choleraesuis.

To identify genes belonging to the Ferric update regulator (Fur) regulon of Salmonella enterica serovar Choleraesuis, the Fur titration assay (FURTA) was used to screen a genomic library for Fur promoters and iron-regulated genes. Fifteen FURTA positive clones were identified from this assay. DNA sequence analysis of these clones showed that 11 out of 15 clones had a Fur binding site (Fur box), and 6 of these clones showed homology to the iron-regulated genes of S. enterica serovar Typhi and/or E. coli. One of these clones (pSC4) was homologous to the iroB gene of the iroA locus of S. enterica serovar Typhi. The iroA locus of S. enterica serovar Choleraesuis was cloned from a lambda-dash library and subjected to DNA sequencing. The complete nucleotide sequence of 9848 bp of the iroA locus of S. enterica serovar Choleraesuis consists of iroB, C, D, E and N genes, which are transcriptionally regulated by Fur. The amino acid sequence of IroB, C, D, E and N was 95%, 86, 89, 96 and 96% identity to that of S. enterica serovar Typhi. The IroN gene was homologous to the family of TonB-dependent outer membrane receptors and the putative virulence factor, IroNE. coli, of the extraintestinal pathogenic E. coli. The convalescent porcine sera contained antibodies against the three major iron-regulated outer membrane proteins of S. enterica serovar Choleraesuis. An insertional inactivation of the iroN gene of S. enterica serovar Choleraesuis by allelic exchange resulted in the loss of expression of the 78 kDa protein. However, this mutant had a similar LD50 to mice compared to the parent strain when given intraperitoneally.