BACKGROUND: About one third of patients with epilepsy become refractory to therapy despite receiving adequate medical treatment, possibly from multidrug resistance. P-glycoprotein, encoded by multidrug resistance protein-1 (MDR1) gene, at the blood brain barrier is considered as a major factor mediating drug efflux and contributing to resistance. Given that peripheral blood mononuclear cells (PBMNCs) express MDR1, we investigated a MDR1 status of PBMNCs in various subsets of epilepsy patients and demonstrated their association with clinical characteristics. METHODOLOGY/PRINCIPAL FINDINGS: Clinical and MDR1 data were collected from 140 patients with epilepsy, 30 healthy volunteers, and 20 control patients taking anti-epileptic drugs. PBMNCs were isolated, and basal MDR1 levels and MDR1 conformational change levels were measured by flow cytometry. MDR1 profiles were analyzed according to various clinical parameters, including seizure frequency and number of medications used in epilepsy patients. Epilepsy patients had a higher basal MDR1 level than non-epilepsy groups (p<0.01). Among epilepsy patients, there is a tendency for higher seizure frequency group to have higher basal MDR1 level (p = 0.059). The MDR1 conformational change level was significantly higher in the high-medication-use group than the low-use group (p = 0.028). Basal MDR1 (OR = 1.16 [95% CI: 1.060-1.268]) and conformational change level (OR = 1.11 [95% CI: 1.02-1.20]) were independent predictors for seizure frequency and number of medications, respectively. CONCLUSIONS/SIGNIFICANCE: The MDR1 profile of PBMNCs is associated with seizure frequency and medication conditions in patients with epilepsy.

BACKGROUND Activated endothelial cells release plasma membrane submicron vesicles expressing CD62E (E-selectin) into blood, known as endothelial microparticles (EMPs). We studied whether the levels of endothelial microparticles expressing CD62E(+), CD31(+)/Annexin-V(+), or CD31(+)/CD42(-) predict cardiovascular outcomes in patients with stroke history. METHODS/PRINCIPAL FINDINGS Patients with stroke history at least 3 months prior to enrolment were recruited. Peripheral blood EMP levels were measured by flow cytometry. Major cardiovascular events and death were monitored for 36 months. Three hundred patients were enrolled, of which 298 completed the study according to protocol. Major cardiovascular events occurred in 29 patients (9.7%). Nine patients died, five from cardiovascular causes. Cumulative event-free survival rates were lower in patients with high levels of CD62E(+) microparticles. Multivariate Cox regression analysis adjusted for cardiovascular risk factors, medications and stroke etiologic groups showed an association between a high CD62E(+) microparticle level and a risk of major cardiovascular events and hospitalization. Levels of other kinds of EMPs expressing CD31(+)/Annexin-V(+) or CD31(+)/CD42(-) markers were not predictive of cardiovascular outcomes. CONCLUSION A high level of CD62E(+) microparticles is associated with cardiovascular events in patients with stroke history, suggesting that the systemic endothelial activation increases the risk for cardiovascular morbidities.

Study Design. We evaluated the influence of rabbit notochordal cells on the expression of inflammatory mediators by human annulus fibrosus (AF) cells co-cultured with macrophage-like cells.Objective. To identify the protective effect of rabbit notochordal cells on AF during in vitro inflammation.Summary of Background Data. Discogenic pain, which is an important cause of intractable lower back pain, is associated with macrophage-mediated inflammation in the AF. Although rabbit notochordal cells prevent intervertebral disc degeneration, their effects on human AF inflammation remain unknown.Methods. Human AF pellets were co-cultured for 48 hours with notochordal cell clusters from adult New Zealand White rabbits and phorbol myristate acetate (PMA)-stimulated human macrophage-like THP-1 cells. Conditioned media (CM) from the co-cultures were assayed by ELISA. The expression of inflammatory mediators in the AF pellets was evaluated by real-time reverse-transcription PCR.Results. The levels of mRNA for IL-6, IL-8, and iNOS in the AF pellets co-cultured with notochordal cells and macrophages [hAF(rNC-M)] were significantly lower than those in the AF pellets cultured with macrophages alone [hAF(M)](P<0.05). The levels of IL-6 and IL-8 protein in the CM of hAF(rNC-M) were significantly lower than those in the CM of hAF(M)(P<0.05). Co-culturing with notochordal cells significantly decreased the levels of mRNA for IL-6, IL-8, and iNOS in the macrophage-exposed AF pellets (P<0.05). Following 1 ng/ml IL-1β stimulation, the levels of IL-6 and IL-8 mRNA and the level of IL-8 protein production were decreased significantly in the AF pellets with notochordal cells compared with naïve AF pellets (P<0.05).Conclusions. In an in vitro co-culture system, rabbit notochordal cells reduced the levels of main inflammatory mediators and gene expression in the human AF during inflammation. Therefore, rabbit notochordal cells may constitute an important protective tool against symptomatic disc development.

Objective: To compare the underestimation rate of invasive carcinoma in cases with ductal carcinoma in situ (DCIS) at percutaneous ultrasound-guided core biopsies of breast lesions between 14-gauge automated core-needle biopsy (ACNB) and 8- or 11-gauge vacuum-assisted biopsy (VAB), and to determine the relationship between the lesion type (mass or microcalcification on radiological findings) and the DCIS underestimation rate.Methods: We retrospectively reviewed imaging-guided biopsies of breast lesions performed from February 2003 to August 2008. 194 lesions were diagnosed as DCIS at ultrasound-guided core biopsy: 138 lesions in 132 patients by 14-gauge ACNB, and 56 lesions in 56 patients by 8- or 11-gauge VAB. The histological results of the core biopsy samples were correlated with surgical specimens. The clinical and radiological findings were also reviewed. The histological DCIS underestimation rates were compared between the two groups and were analysed for differences according to the clinical and radiological characteristics of the lesions.Results: The DCIS underestimation rate was 47.8%(66/138) for 14-gauge ACNB and 16.1%(9/56) for VAB (p<0.001). According to the lesion type on sonography, DCIS underestimation was 43.4%(63/145) in masses (47.6% using ACNB and 15.8% using VAB; p = 0.012) and 24.5%(12/49) in microcalcifications (50.0% using ACNB and 16.2% using VAB; p = 0.047).Conclusion: The underestimation rate of invasive carcinoma in cases with DCIS at ultrasound-guided core biopsies was significantly higher for ACNB compared with that for VAB. Furthermore, this difference does not change according to the lesion type on ultrasound. Therefore, ultrasound-guided VAB can be a useful method for the diagnosis of DCIS lesions presented as either mass or microcalcification.

In addition to development of vaccines and synthetic antiviral drugs, recent studies have advocated the use of natural substances that inhibit or prevent viral infections. High-molecular-weight poly-γ-glutamate (HM-γ-PGA) produced by Bacillus subtilis chungkookjang was evaluated for anti-influenza virus activity. HM-γ-PGA induced type I interferons (IFNs), which in turn stimulated expression of Myxovirus resistant 1 protein and IFN-related proteins in vitro. In the B6.A2G-Mx1 mouse model, which mimics the innate immune system of humans, treatment with HM-γ-PGA enhanced the antiviral state of mice and protected them against highly pathogenic influenza A virus. Naturally synthesized HM-γ-PGA has potent anti-influenza activity and may be a useful means for control of influenza virus.

BACKGROUND: The purpose of this study was to investigate the clinical and ultrasonographic (US) findings affecting the nondiagnostic cytology and malignancy rate in thyroid nodules with nondiagnostic cytology upon a second ultrasound-guided fine needle aspiration (US-FNA). MATERIALS AND METHODS: A total of 1124 nodules in 1081 patients (M:F = 111:970; mean age, 48.5 years) with 2 or more US-FNAs were included. Clinical and US features were compared between the nodules with nondiagnostic results and diagnostic results from their second FNA using univariate and multivariate analyses. The malignancy rate was analyzed in terms of nondiagnostic cytology. RESULTS: Among 458 nondiagnostic results of the initial FNA, 94 nodules (20.5%) still had nondiagnostic results on the second FNA. Having cystic portion >50%(odds ratio [OR] = 2.845, P = .0001), nondiagnostic cytology of the first FNA (OR = 2.813, P < .0001), a nodule size ≤5 mm (OR = 1.742, P = .0331), or hypoechogenicity (OR = 1.512, P = .0404) were factors significantly affecting the incidence of nondiagnostic results in the second US-FNA based on multivariate analysis. The malignancy rate was 11.4% in the nodules with nondiagnostic results in the first FNA and 3.2% in the nodules with serial nondiagnostic results in both the first and second FNAs. CONCLUSIONS: The high probability of nondiagnostic results from a second FNA should be considered if a nodule shows cystic portion greater than 50%, nondiagnostic cytology on the first FNA, a nodule size ≤5 mm, or hypoechogenicity. The malignancy rate of thyroid nodules with 2 consecutive nondiagnostic results was 3.2%.

Safflower seed has been reported to have a protective effect against bone loss diseases. However, the precise molecular mechanisms underlying the inhibitory effect of safflower seed in osteoclast differentiation remain unclear. In this study, we investigated the inhibitory action of safflower seed extract (SSE) on the receptor activator of nuclear factor κB ligand (RANKL)-induced osteoclastogenesis in cultured mouse-derived bone marrow macrophages (BMMs). We found that SSE significantly inhibited the formation of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells in BMMs without cytotoxicity. The gene expressions of nuclear factor of activated T-cells (NFATc1) and TRAP, which are genetic markers of osteoclast differentiation, were substantially decreased by SSE in a dose-dependent manner. Also, SSE diminished RANKL-mediated intracellular reactive oxygen species (ROS) generation on osteoclastogenesis in a dose-dependent manner. The SSE thereafter suppressed RANKL-induced p38 mitogen-activated protein kinase and IκBα kinase signalling activities which were activated by ROS generation for osteoclastogenesis. Additionally, SSE was found to decrease RANKL-induced actin ring formation, which is required for bone resorption activity. Taken together, our results suggest that SSE acts as a RANKL-induced osteoclastogenesis inhibitor by suppression of ROS generation. This induces a remarkable suppression of the p38 and IκBα kinase pathways, thereby suppressing the gene expression of NFATc1 in osteoclast precursors. Copyright © 2012 John Wiley & Sons, Ltd.

Purpose: To evaluate the diagnostic performance of gray-scale ultrasonography (US) and elastography in differentiating benign and malignant thyroid nodules. Materials and Methods: This was an institutional review board-approved retrospective study with waiver of informed consent. A total of 703 solid thyroid nodules in 676 patients (mean age, 49.7 years; range, 18-79 years) were included; there were 556 women (mean age, 49.5 years; range, 20-74 years) and 120 men (mean age, 50.7 years; range, 18-79 years). Nodules with marked hypoechogenicity, poorly defined margins, microcalcifications, and a taller-than-wide shape were classified as suspicious at grayscale US. Findings at elastography were classified according to the Rago criteria and the Asteria criteria. The diagnostic performances of gray-scale US and elastography were compared. For comparison between the diagnostic performances of gray-scale US and the combination of gray-scale US and elastography, three sets of criteria were assigned: criteria set 1, nodules with any suspicious grayscale US feature were assessed as suspicious; criteria set 2, Rago criteria were added as suspicious features to criteria set 1; and criteria set 3, Asteria criteria were added as suspicious features to criteria set 1. The diagnostic performances of gray-scale US, elastography with Rago criteria, and elastography with Asteria criteria, and odds ratios (ORs) with 95% confidence intervals for predicting thyroid malignancy were compared using generalized estimating equation analysis. Results: Of 703 nodules, 217 were malignant and 486 were benign. Sensitivity, negative predictive value (NPV), and OR of gray-scale US for the 703 nodules were 91.7%, 94.7%, and 22.1, respectively, and these values were higher than the 15.7% and 65.4% sensitivity, 71.7% and 79.1% NPV, and 3.7 and 2.6 ORs found for elastography with Rago and Asteria criteria, respectively. Specificity, positive predictive value, and accuracy for criteria set 1 were significantly higher than those for criteria sets 2 and 3 for most of the nodule subgroups that were considered. Conclusion: Elastography alone, as well as the combination of elastography and gray-scale US, showed inferior performance in the differentiation of malignant and benign thyroid nodules compared with gray-scale US features; elastography was not a useful tool in recommending fine-needle aspiration biopsy. © RSNA, 2012.

Ribitol dehydrogenase from Zymomonas mobilis (ZmRDH) catalyzes the conversion of ribitol to d-ribulose and concomitantly reduces NAD(P)(+) to NAD(P)H. A systematic approach involving an initial sequence alignment-based residue screening, followed by a homology model-based screening and site-directed mutagenesis of the screened residues, was used to study the molecular determinants of the cofactor specificity of ZmRDH. A homologous conserved amino acid, Ser156, in the substrate-binding pocket of the wild-type ZmRDH was identified as an important residue affecting the cofactor specificity of ZmRDH. Further insights into the function of the Ser156 residue were obtained by substituting it with other hydrophobic nonpolar or polar amino acids. Substituting Ser156 with the negatively charged amino acids (Asp and Glu) altered the cofactor specificity of ZmRDH toward NAD(+)(S156D,[k(cat)/K(m)(,NAD)]/[k(cat)/K(m)(,NADP)]= 10.9, where K(m)(,NAD) is the K(m) for NAD(+) and K(m)(,NADP) is the K(m) for NADP(+)). In contrast, the mutants containing positively charged amino acids (His, Lys, or Arg) at position 156 showed a higher efficiency with NADP(+) as the cofactor (S156H,[k(cat)/K(m)(,NAD)]/[k(cat)/K(m)(,NADP)]= 0.11). These data, in addition to those of molecular dynamics and isothermal titration calorimetry studies, suggest that the cofactor specificity of ZmRDH can be modulated by manipulating the amino acid residue at position 156.

The aim was to compare the diagnostic performances of physicians with different levels of experience for completing preoperative staging ultrasonography (US) for thyroid malignancy. From March 2009 to December 2009, 1421 patients underwent preoperative staging US by six physicians according to the sixth edition of TNM classification. Extrathyroidal extension was defined as a perimeter in contact with >25% of the thyroid capsule in a malignant lesion or loss of the capsule line. US features in diagnosis of lymph node metastases were round shape, hyperechogenicity, microcalcifications, cystic change, and peripheral vascularity. Diagnostic performances of the three experienced physicians (>7 y) and the three less experienced physicians (<2 y) were compared. The overall diagnostic performances of T, N1a, multifocality and bilaterality by the experienced and less experienced physicians were comparable. The positive predictive value of N1b by experienced physicians was 64%, whereas that by less experienced physicians was 38.5%. These results suggest that evaluation of lateral lymph node metastasis requires more experience than other aspects of staging US for thyroid malignancy.