Dachshund Homolog 1: Unveiling Its Potential Role in Megakaryopoiesis and Bacillus anthracis Lethal Toxin-Induced Thrombocytopenia.

Lethal toxin (LT) is the critical virulence factor of Bacillus anthracis, the causative agent of anthrax. One common symptom observed in patients with anthrax is thrombocytopenia, which has also been observed in mice injected with LT. Our previous study demonstrated that LT induces thrombocytopenia by suppressing megakaryopoiesis, but the precise molecular mechanisms behind this phenomenon remain unknown. In this study, we utilized 12-O-tetradecanoylphorbol-13-acetate (TPA)-induced megakaryocytic differentiation in human erythroleukemia (HEL) cells to identify genes involved in LT-induced megakaryocytic suppression. Through cDNA microarray analysis, we identified Dachshund homolog 1 (DACH1) as a gene that was upregulated upon TPA treatment but downregulated in the presence of TPA and LT, purified from the culture supernatants of B. anthracis. To investigate the function of DACH1 in megakaryocytic differentiation, we employed short hairpin RNA technology to knock down DACH1 expression in HEL cells and assessed its effect on differentiation. Our data revealed that the knockdown of DACH1 expression suppressed megakaryocytic differentiation, particularly in polyploidization. We demonstrated that one mechanism by which B. anthracis LT induces suppression of polyploidization in HEL cells is through the cleavage of MEK1/2. This cleavage results in the downregulation of the ERK signaling pathway, thereby suppressing DACH1 gene expression and inhibiting polyploidization. Additionally, we found that known megakaryopoiesis-related genes, such as FOSB, ZFP36L1, RUNX1, FLI1, AHR, and GFI1B genes may be positively regulated by DACH1. Furthermore, we observed an upregulation of DACH1 during in vitro differentiation of CD34-megakaryocytes and downregulation of DACH1 in patients with thrombocytopenia. In summary, our findings shed light on one of the molecular mechanisms behind LT-induced thrombocytopenia and unveil a previously unknown role for DACH1 in megakaryopoiesis.

Recombinant Klotho attenuates IFNγ receptor signaling and SAMHD1 expression through blocking NF-κB translocation in glomerular mesangial cells.

Interferon gamma (IFNγ) is a cytokine implicated in the pathogenesis of autoimmune diseases. SAM and HD domain-containing protein 1 (SAMHD1) is an IFNγ-inducible protein that modulates cellular dNTP levels. Mutations in the human SAMHD1 gene cause Aicardi-Goutières (AG) syndrome, an autoimmune disease sharing similar clinical features with systemic lupus erythematosus (SLE). Klotho is an anti-inflammatory protein which suppresses aging through multiple mechanisms. Implication of Klotho in autoimmune response is identified in rheumatologic diseases such as SLE. Little information exists regarding the effect of Klotho in lupus nephritis, one of the prevalent symptoms of SLE. The present study verified the effect of IFNγ on SAMHD1 and Klotho expression in MES-13 glomerular mesangial cells, a special cell type in glomerulus that is critically involved in lupus nephritis. IFNγ upregulated SAMHD1 expression in MES-13 cells through the Janus kinase-signal transducer and activator of transcription 1 (JAK-STAT1) and the nuclear factor kappa B (NFκB) signaling pathways. IFNγ decreased Klotho protein expression in MES-13 cells. Treatment of MES-13 cells with recombinant Klotho protein inhibited SAMHD1 expression by blocking IFNγ-induced NFκB nuclear translocation, but showed no effect on JAK-STAT1 signaling. Collectively, our findings support the protective role of Klotho in attenuating lupus nephritis through the inhibition of IFNγ-induced SAMHD1 expression and IFNγ downstream signaling in MES-13 cells.

Rapid and sensitive detection of Yersinia pestis by lateral-flow assay in simulated clinical samples.

BACKGROUND: Yersinia pestis is a contributing agent to the epidemic disease, plague, which killed an estimated 200 million people during historical times. In this study, a rapid, cheap, sensitive, and specific technique, the lateral flow assay (F1 strips), has been successfully developed to detect this pathogen, by using paired monoclonal antibodies (MAbs) against Y. pestis capsule like fraction 1 (F1) protein. Compared with the polyclonal antibody (PAb) based F1 strips, the Mab-based F1 strips have a remarkable increased detection limitation (10 to 100 folds). Furthermore, besides the limitation and specificity evaluation, the application of this F1 strip on simulated clinical samples indicate the LFA can be a good candidate to detect plague. METHODS: Recombinant F1 antigen was expressed and purified from a series of works. The various anti-F1 monoclonal antibodies generated from hybridoma cells were screened with the ELISA technique. To evaluate the feasibility of this Y. pestis F1 test strip, the F1 protein/Y. pestis was spiked into simulated clinical samples such as human serum, mouse bronchoalveolar lavage fluids, and mouse blood to mimic natural infection status. Additionally, this technique was applied to detect the Y. pestis in the environment-captured rats, to evaluate the practical usefulness of the strips. RESULTS: By using this MAb-based-LFA technique, 4 ng/ml of recombinant F1-protein and 103 CFU/ml of Y. pestis could be detected in less than 10 mins, which is at least 10-folds than that of the PAb format. On the other hand, although various Yersinia strains were applied to the strips, only Y. pestis strain showed a positive result; all other Yersinia species did not produce a positive signal, indicating the high efficiency and specificity of the MAb-based F1-strips. CONCLUSION: Based on our findings, we suggest that the MAb-format-LFA will be valuable as a diagnostic tool for the detection of Y. pestis. This report shows that the F1 strip is sufficient to support not only the detection of plague in simulated clinical samples, but also it may be a good candidate to meet the epidemiological surveillance during an outbreak of the biological warfare.

Effect of Age on Working Memory Performance and Cerebral Activation after Mild Traumatic Brain Injury: A Functional MR Imaging Study.

PURPOSE: To evaluate the age effect on working memory (WM) performance and functional activation after mild traumatic brain injury (MTBI). MATERIALS AND METHODS: This study was approved by the local research ethics committee. All participants provided written informed consent. N-back WM cerebral activation was assessed with functional magnetic resonance (MR) imaging in 13 younger (mean age, 26.2 years ± 2.9; range, 21-30 years) and 13 older (mean age, 57.8 years ± 6.6; range, 51-68 years) patients with MTBI and 26 age- and sex-matched control subjects. Two functional MR images were obtained within 1 month after injury and 6 weeks after the initial study. Group comparison and regression analysis were performed among postconcussion symptoms, neuropsychologic tests, and WM activity in both groups. RESULTS: In younger patients, initial hyperactivation was seen in the right precuneus and right inferior parietal gyrus (P = .047 and P = .025, respectively) in two-back greater than one-back conditions compared with younger control subjects, whereas in older patients, hypoactivation was seen in the right precuneus and right inferior frontal gyrus (P = .013 and P =.019, respectively) compared with older control subjects. Increased WM activity was associated with increased postconcussion symptoms in the right precuneus (r = 0.57; P = .026) and right inferior frontal gyrus (r = 0.60; P = .019) and poor WM performance in the right precuneus (r = -0.55; P = .027) in younger patients at initial studies but not in older patients. At follow-up examinations, partial recovery of activation pattern and decreased postconcussion symptoms (P = .04) were observed in younger patients but not in older patients. CONCLUSION: The different manifestations of postconcussion symptoms at functional MR imaging between younger and older patients confirmed the important role of age in the activation, modulation, and allocation of WM processing resources after MTBI. These findings also supported that younger patients have better neural plasticity and clinical recovery than do older patients.

Sex Differences in Working Memory after Mild Traumatic Brain Injury: A Functional MR Imaging Study.

PURPOSE: To evaluate sex differences in mild traumatic brain injury (MTBI) with working memory functional magnetic resonance (MR) imaging. MATERIALS AND METHODS: Research ethics committee approval and patient written informed consent were obtained. Working memory brain activation patterns were assessed with functional MR imaging in 30 patients (15 consecutive men and 15 consecutive women) with MTBI and 30 control subjects (15 consecutive men and 15 consecutive women). Two imaging studies were performed in patients: the initial study, which was performed within 1 month after the injury, and a follow-up study, which was performed 6 weeks after the first study. For each participant, digit span and continuous performance testing were performed before functional MR imaging. Clinical data were analyzed by using Kruskal-Wallis, Mann-Whitney U, Wilcoxon signed rank, and Fisher exact tests. Within- and between-group differences of functional MR imaging data were analyzed with one- and two-sample t tests, respectively. RESULTS: Among female participants, the total digit span score was lower in the MTBI group than in the control group (P = .044). In initial working memory functional MR imaging studies, hyperactivation was found in the male MTBI group and hypoactivation was found in the female MTBI group compared with control male and female groups, respectively. At the 6-week follow-up study, the female MTBI group showed persistent hypoactivation, whereas the male MTBI group showed a regression of hyperactivation at visual comparison of activation maps. The male MTBI group was also found to have a higher initial ß value than the male control group (P = .040), and there was no significant difference between the male MTBI group and the male control group (P = .221) at follow-up evaluation, which was comparable to findings on activation maps. In the female MTBI group, average ß values at both initial and follow-up studies were lower compared with those in the female control group but were not statistically significant (P = .663 and P = .191, respectively). CONCLUSION: Female patients with MTBI had lower digit span scores than did female control subjects, and functional MR imaging depicted sex differences in working memory functional activation; hypoactivation with nonrecovery of activation change at follow-up studies may suggest a worse working memory outcome in female patients with MTBI.

May-Thurner syndrome: correlation between digital subtraction and computed tomography venography.

BACKGROUND/PURPOSE: Digital subtraction venography (DSV) and computed tomography venography (CTV) are both recommended for diagnosing May-Thurner syndrome. The literature contains little information on the correlation between these imaging tools. We performed a retrospective case-series study to investigate this correlation. METHODS: From August 2009 to August 2010, 42 patients with May-Thurner syndrome (34 women, 8 men; mean age: 52.8 ± 13.5 years) received DSV followed by CTV. The DSV was used to evaluate the degree of venous reflux, reflux start-up time, and flow time. By CTV, the ratio of cross-sectional area and the ratio of diameter between the narrowest region to that of the caudal part of the left common iliac vein were calculated. The correlation between these variables for DSV versus CTV was calculated using Spearman's rank correlation coefficients. RESULTS: In DSV evaluation of the extent of reflux, 19.0% of cases were classified as Grade 0, 11.9% as Grade I, 28.6% as Grade II, and 40.5% as Grade III. The mean ± standard deviation flow times for these groups were 2.00 ± 0.38 seconds, 1.75 ± 0.29 seconds, 1.67 ± 0.72 seconds, and 1.81 ± 0.68 seconds, the mean time for total patients was 1.76 ± 0.78 seconds. The reflux start-up times for Grades I-III were 2.00 ± 1.00 seconds, 1.80 ± 1.23 seconds, and 1.40 ± 0.49 seconds, and the mean time was 1.6 ± 0.8 seconds. In CTV, the mean area ratio and diameter ratio were 0.78 ± 0.22 (range, 0.22-1.27) and 0.75 ± 0.24 (range, 0.33-1.25). The reflux start-up time showed a positive correlation with the cross-sectional area ratio (r = 0.518; p = 0.002) and diameter ratio (r = 0.413; p = 0.019). CONCLUSION: The cross-sectional area ratio and diameter ratio in CTV correlate with the reflux start-up time in DSV. For May-Thurner syndrome, both CTV and DSV provide essential information for diagnosis and evaluation of the disease. The positive correlation between anatomical and hemodynamic properties corresponds with the underlying pathophysiology.

Structural and functional analyses of a glutaminyl cyclase from Ixodes scapularis reveal metal-independent catalysis and inhibitor binding.

Glutaminyl cyclases (QCs) from mammals and Drosophila are zinc-dependent enzymes that catalyze N-terminal pyroglutamate formation of numerous proteins and peptides. These enzymes have been found to be critical for the oviposition and embryogenesis of ticks, implying that they are possible physiological targets for tick control. Here, 1.10-1.15 Šresolution structures of a metal-independent QC from the black-legged tick Ixodes scapularis (Is-QC) are reported. The structures exhibit the typical scaffold of mammalian QCs but have two extra disulfide bridges that stabilize the central ß-sheet, resulting in an increased thermal stability. Is-QC contains ~0.5 stoichiometric zinc ions, which could be removed by 1 mM EDTA. Compared with the Zn-bound form, apo-Is-QC has a nearly identical active-site structure and stability, but unexpectedly possesses significantly increased QC activities towards both synthetic and physiological substrates. Enzyme-kinetic analysis revealed that apo-Is-QC has a stronger substrate-binding affinity, suggesting that bound zinc interferes with substrate binding during catalysis. The structures of Is-QC bound to the inhibitor PBD150 revealed similar binding modes to both forms of Is-QC, with the exception of the inhibitor imidazole ring, which is consistent with the comparable inhibition activities of the inhibitor towards both forms of Is-QC. These findings have implications for the design of new QC inhibitors.

Suspension bead array of the single-stranded multiplex polymerase chain reaction amplicons for enhanced identification and quantification of multiple pathogens.

Rapid identification of single and multiple infectious agents is vital in clinical settings and during biothreat attack. This study assesses the assay of single-stranded multiplex polymerase chain reaction (PCR) amplicons by suspension bead array (SSMP-SBA) for multiple pathogens identification in a single-tube reaction. A 15-plex assay for identification of 11 highly infectious pathogens was developed to evaluate the performance of SSMP-SBA. Pathogen-specific amplicons were obtained by sequential amplification of genomic DNAs using gene-specific primers tagged with artificial unique sequences and unique primers of which the reverse primer was modified by biotin and phosphorothioate. The SSMP products generated by T7 exonuclease-mediated DNA hydrolysis were hybridized to 15 sets of beads coupled with gene-specific and control oligonucleotide probes for pathogen identification and quantification by flow cytometry. This method was validated via assessment of 57 reference strains and one clinical bacterial isolate. All 11 pathogens can be detected by the 15-plex SSMP-SBA assay, and this design significantly enhanced the signal-to-noise ratio and improved the assay performance. This assay achieves similar sensitivity to our in-house real-time PCR system with the limit of detection equivalent to 5-100 genome copies and a linear dynamic range crossing three to five logs. In the validation assay, a 100% accuracy rate was achieved when the pathogens were among the target species. Notably, the species of pathogens were accurately identified from the samples with multiple infections. SSMP-SBA presents superior performance with multiplexing capability in a single-tube reaction and provides a new approach for detection and species identification of multiple pathogen infections.

Working memory in patients with mild traumatic brain injury: functional MR imaging analysis.

PURPOSE: To analyze brain activation patterns in response to tests of working memory after a mild traumatic brain injury (MTBI). MATERIALS AND METHODS: Research ethics committee approval and patient written informed consent were obtained. Brain activation patterns in response to n-back working memory tasks (n = 1, 2, 3) were assessed with functional magnetic resonance (MR) imaging in 20 patients with MTBI within 1 month after their injury and in 18 healthy control subjects. In n-back working memory tasks, participants monitored a series of number stimuli and were to indicate when the presented number was the same as that presented n back previously. Nine (45%) MTBI patients underwent follow-up functional MR imaging studies 6 weeks later. Digit span, a memory test for how many numbers a person can remember in sequence, and continuous performance test (CPT), a test that measures a person's sustained and selective attention and impulsivity, were also performed before functional MR imaging studies and outside the imager for each participant. Clinical data were analyzed by using t and χ(2) tests. Within-group, between-group, and initial and follow-up differences of functional MR imaging data were analyzed by using one-sample, two-sample, and paired t tests, respectively. RESULTS: Groups were similar for sex (P = .75), years of education (P = .069), digit span (P = .37 for total score), CPT (P = .31, .27, and .43 for omission error, commission error, and hit reaction time, respectively), and accuracy of n-back working memory performance (P = .90, .11, and .39 for one-, two-, and three-back tasks, respectively). Brain activation patterns differed between MTBI patients and controls in response to increasing working memory loads (P < .01, uncorrected). Control subjects maintained their ability to increase activation in the working memory circuitry with each increase in working memory load. In contrast, MTBI patients were impaired in their ability to increase activation in working memory circuitry under both moderate and high working memory load conditions. However, MTBI patients did show cerebral plasticity, as evidenced by more activation in some areas outside and inside the working memory circuitry as compared with control subjects (P < .01, uncorrected). In the 6-week follow-up study, compared with baseline, MTBI patients showed an improvement of activation in response to increasing working memory loads (P < .05, uncorrected). CONCLUSION: MTBI-induced differences in working memory functional activity were observed even though differences in behavioral performance between MTBI patients and controls were absent, which suggests that this approach may increase sensitivity to MTBI compared with neuropsychological evaluation alone.

Methotrexate inhibition of SARS-CoV-2 entry, infection and inflammation revealed by bioinformatics approach and a hamster model.

Background: Drug repurposing is a fast and effective way to develop drugs for an emerging disease such as COVID-19. The main challenges of effective drug repurposing are the discoveries of the right therapeutic targets and the right drugs for combating the disease. Methods: Here, we present a systematic repurposing approach, combining Homopharma and hierarchal systems biology networks (HiSBiN), to predict 327 therapeutic targets and 21,233 drug-target interactions of 1,592 FDA drugs for COVID-19. Among these multi-target drugs, eight candidates (along with pimozide and valsartan) were tested and methotrexate was identified to affect 14 therapeutic targets suppressing SARS-CoV-2 entry, viral replication, and COVID-19 pathologies. Through the use of in vitro (EC50 = 0.4 µM) and in vivo models, we show that methotrexate is able to inhibit COVID-19 via multiple mechanisms. Results: Our in vitro studies illustrate that methotrexate can suppress SARS-CoV-2 entry and replication by targeting furin and DHFR of the host, respectively. Additionally, methotrexate inhibits all four SARS-CoV-2 variants of concern. In a Syrian hamster model for COVID-19, methotrexate reduced virus replication, inflammation in the infected lungs. By analysis of transcriptomic analysis of collected samples from hamster lung, we uncovered that neutrophil infiltration and the pathways of innate immune response, adaptive immune response and thrombosis are modulated in the treated animals. Conclusions: We demonstrate that this systematic repurposing approach is potentially useful to identify pharmaceutical targets, multi-target drugs and regulated pathways for a complex disease. Our findings indicate that methotrexate is established as a promising drug against SARS-CoV-2 variants and can be used to treat lung damage and inflammation in COVID-19, warranting future evaluation in clinical trials.

Attributable mortality burden of metabolic syndrome: comparison with its individual components.

AIM: To estimate the national prevalence, mortality risk and population mortality burden of metabolic syndrome, and compare the values with those of its individual components. METHODS AND RESULTS: A total of 486,341 apparently healthy adults who went through a screening programme in Taiwan were recruited from 1994 onwards. As of 2007, 15,268 deaths had occurred at least one year after the examination. Six definitions of metabolic syndrome were used. Components of metabolic syndrome include obesity, hypertension, hyperglycaemia, dyslipidaemia and albuminuria. Hazard ratios (HRs) were calculated using the Cox proportional hazard model. The population mortality burden considered both national prevalence and HRs. The national prevalence of metabolic syndrome defined by the Adult Treatment Panel (ATP) III was 16.3%, the HR for all causes was 1.36 (95%, CI 1.31-1.41) and the HR for cardiovascular disease (CVD) was 1.63 (95%, CI 1.51-1.77). The population mortality burden of metabolic syndrome was 5.5% for all causes, in contrast to 9.0% for hypertension, 8.9% for albuminuria, 6.6% for diabetes, 3.5% for dyslipidaemia and 1.5% for obesity. For CVD it was 9.4%, lower than 10.7% for albuminuria and 25.0% for hypertension. CONCLUSION: The mortality burden of metabolic syndrome was relatively small at national level. Three of the five components of metabolic syndrome alone, namely hypertension, diabetes and albuminuria, contributed more than metabolic syndrome to all-cause mortality. Successful management of any of these three components would have achieved a greater impact on mortality than management of metabolic syndrome.

A Unique Carboxylic-Acid Hydrogen-Bond Network (CAHBN) Confers Glutaminyl Cyclase Activity on M28 Family Enzymes.

Proteins with sequence or structure similar to those of di-Zn exopeptidases are usually classified as the M28-family enzymes, including the mammalian-type glutaminyl cyclases (QCs). QC catalyzes protein N-terminal pyroglutamate formation, a posttranslational modification important under many physiological and pathological conditions, and is a drug target for treating neurodegenerative diseases, cancers and inflammatory disorders. Without functional characterization, mammalian QCs and their orthologs remain indistinguishable at the sequence and structure levels from other M28-family proteins, leading to few reported QCs. Here, we show that a low-barrier carboxylic-acid hydrogen-bond network (CAHBN) is required for QC activity and discriminates QCs from M28-family peptidases. We demonstrate that the CAHBN-containing M28 peptidases deposited in the PDB are indeed QCs. Our analyses identify several thousands of QCs from the three domains of life, and we enzymatically and structurally characterize several. For the first time, the interplay between a CAHBN and the binuclear metal-binding center of mammalian QCs is made clear. We found that the presence or absence of CAHBN is a key discriminator for the formation of either the mono-Zn QCs or the di-Zn exopeptidases. Our study helps explain the possible roles of QCs in life.

Cancer risks from betel quid chewing beyond oral cancer: a multiple-site carcinogen when acting with smoking.

OBJECTIVES: This cohort study is to assess the extent of cancer risks of betel quid chewing (without tobacco added) beyond oral cancer, as such information was limited from case-control studies. METHODS: The cohort, selected from participants in a medical screening program since 1994, consisted of 177,271 adult men with 19.2% chewers of betel quid. As of 2006, out of 4,840 deaths, 1,901 cancer deaths were identified. Mortality hazard ratios (HR) were estimated by Cox proportional hazard model. Life expectancy was calculated by life table method. RESULTS: One-third of smokers chewed (33%) but most of chewers smoked (90%). Risk for all cancer doubled among chewers (HR = 2.00). Risks of at least six cancer sites were increased among chewers: oral cavity (HR = 12.52), esophagus (HR = 5.64), liver (HR = 2.27), pancreas (HR = 2.67), larynx (HR = 6.24), and lung (HR = 2.43) with risks increased with increasing betel quid amount consumed. All-cancer age-adjusted mortality rates in Taiwan increased 25%, including 223% increase in oral cancer, during the last 20 years when chewing rate increased five- to tenfolds. Chewing on top of smoking increased the risks synergistically, and these two were responsible for at least half (50%) of all cancer deaths among 2 million chewers in Taiwan. Life expectancy of chewers was shorter than non-chewers by 5.93 years at age 20 and 5.55 years at age 40. CONCLUSION: In addition to oral cancer, significant increases were seen among chewers for cancer of the esophagus, liver, pancreas, larynx, lung, and all cancer. Chewing and smoking, as combined by most chewers, interacted synergistically and was responsible for half of all cancer deaths in this group. They were responsible for the recent increases in oral, esophageal, pancreatic, and liver cancer in Taiwan. Chewing and smoking shortened their life span by nearly 6 years.

Role of amphotericin B upon enhancement of protective immunity elicited by oral administration with liposome-encapsulated-Japanese encephalitis virus nonstructural protein 1 (NS1) in mice.

Amphotericin B (AmB) is an antifungal antibiotic the activity of which has been associated with modulation of pro-inflammatory cytokines expression in cultured cells. Herein we reveal that co-administration with AmB enhances the immunogenicity of oral Lip-JENS1 vaccine which derived from liposomes functionalized with DSPC (distearoylphosphatidylcholine) and cholesterol (2:1, molar ratio)-bearing JE virus NS1 protein (600 microg ml(-1)). Oral single dose of Lip-JENS1 elicited a detectable serum NS1-specific IgG antibody response from a mouse model. Remarkably, the addition of AmB (125 microg per mouse), particularly, 2 h prior to, but not simultaneously with, the administration of Lip-JENS1 significantly enhanced the systemic antigen-specific antibody response, providing superior protection against lethal JEV challenges. Further, we observed AmB-induced the transcription of cytokine expression and translocation of transcriptional factor NF-kappaB from the cytoplasm to the nucleus for the murine macrophage J774A.1. Moreover, Peyer's-patch lymphocytes (PPL) from AmB-treated mice produced high levels of IL-1beta, IL-6 and TNF-alpha expression compared to the corresponding control of cells from non-treated mice. Taken together, the results suggest that AmB exerts a profound influence upon mucosal vaccination with Lip-JENS1, possibly playing an adjuvant-augmented role to "fine-tune" humoral as well as cellular immune response, thus conferring enhanced protective immunity for immunising individuals against JE infection.

Electrorheological Sensor Encapsulating Microsphere Media for Plague Diagnosis with Rapid Visualization.

Plague is a disease infected by an etiological agent, which is transmitted from fleas to a variety of wildlife rodents. Therefore, rapid diagnosis of plague on-site in the field is important. Polystyrene microspheres (SMs) of 2.2 µm diameter were synthesized by emulsion polymerization to adsorb magnetic nanoparticles (FNs), resulting in core-/shell-structured microspheres that generate a significant contrast in relative permittivities between SMs and FNs. Electrorheological displays (EDs) consisting of two indium tin oxide glasses with spacers were constructed to contain core-/shell-structured SM/FN (SM@FN) solutions for observing their transmittance change. The ED encapsulating dispersed SM@FN solution exhibited an opaque state because light was scattered significantly without the application of an alternating electric field (AEF). In the presence of an AEF, the particle chaining behavior results in enhancement of the transmittance of ED. At a specific frequency, the so-called characteristic frequency (Fc), the transmittance reaches a maximum. Fc could be used as an indicator to mark the shell materials. The antibody of Yersinia pestis (ab-Yp) was coated onto the SM@FN as a biosensing medium. The Fc of ab-Yp-modified microspheres shifted from 200 to 750 kHz with antigen coupling of Y. pestis antigen (ag-Yp). In the absence of fluorescence labeling, the large change in ED transmittance could be visualized during the Y. pestis detection. The limit of detection and the limit of quantification were ∼30 and ∼40 ng/µL, respectively, obtained within 30 s according to the highest transmittance of ED under the AEF at 750 kHz. Y. pestis detection was not affected by Escherichia coli and Staphylococcus aureus significantly. Compared with other common immunoassays, including the secondary immunochemical or enzyme-linked steps, this simple electrorheological sensor with high sensitivity and selectivity could be a candidate for on-site plague diagnosis.

Increased epicardial adipose tissue (EAT) volume in type 2 diabetes mellitus and association with metabolic syndrome and severity of coronary atherosclerosis.

OBJECTIVE: Epicardial adipose tissue (EAT) is a part of visceral fat deposited around the heart between the pericardium and myocardium along the distribution of coronary arteries. EAT thickness is reported to be associated with coronary atherosclerosis; however, no study has measured EAT volume in patients with type 2 diabetes or investigate its association with coronary artery disease. DESIGN: A hospital-based case control study. PATIENTS: A total of 49 patients with type 2 diabetes mellitus (T2DM) and 78 nondiabetic controls were studied. MEASUREMENTS: Cardiac multislice computed tomography was used to measure EAT volume, Gensini score, coronary artery calcium score and, coronary lesions. The relationships between EAT volume, markers of coronary atherosclerosis and anthropometric and biochemical parameters of metabolic syndrome (MetS) were investigated. RESULTS: EAT volume was significantly higher in patients with T2DM than in nondiabetic subjects (166.1 +/- 60.6 cm(3) vs. 123.4 +/- 41.8 cm(3), P < 0.0001). Logistic regression analysis revealed independent and significant associations between EAT and diabetic status. EAT volume was significantly associated with components of MetS (BMI, waist circumference, fasting serum glucose, total cholesterol, HDL-cholesterol, and triglycerides levels), Gensini score, coronary lesions, coronary disease and coronary calcium scores. Univariate, multivariate and trend analyses confirmed that EAT volume was associated with MetS component clustering and the coronary atherosclerosis index. CONCLUSIONS: The analytical results indicate that EAT volume is increased in T2DM patients and is associated with unfavourable components of MetS and coronary atherosclerosis. The close anatomical relationship between EAT and the coronary arteries, combined with other evidence indicating that EAT is a biologically active adipokine-secreting tissue, suggest that EAT participates in the pathogenesis of diabetic coronary atherosclerosis.

Prediction of cerebral hyperperfusion syndrome after carotid stenting: a cerebral perfusion computed tomography study.

PURPOSE: The objective of this study was to evaluate the role of cerebral perfusion computed tomography (CT) in predicting cerebral hyperperfusion syndrome (CHS) after carotid stenting. MATERIALS AND METHODS: This study was approved by the institutional review board, and written informed consent was obtained from all patients. Fifty-five consecutive symptomatic patients with greater than or equal to 70% of cervical carotid artery stenosis who underwent carotid stenting from March 2001 to December 2003 were recruited. Age, sex, stenting side, and degree of cervical carotid stenosis at the stenting and contralateral sides were recorded. Cerebral perfusion CT was performed 1 day before stenting. Absolute values of the cerebral blood volume, mean transit time, and cerebral blood flow were calculated. Relative values based on the comparison between ipsilateral and contralateral hemispheres, that is, relative cerebral blood volume (ipsilateral-to-contralateral ratio), relative cerebral blood flow (ipsilateral-to-contralateral ratio), and absolute difference in mean transit time (dMTT), were derived. The association between occurrence of CHS and parameters of cerebral perfusion CT was investigated by the Mann-Whitney U test. RESULTS: Three (5%) of 55 patients had CHS after carotid stenting. The only significant factor related to the occurrence of CHS was dMTT (P = 0.003). A dMTT value of 3 seconds was considered as a cutoff value to distinguish between the occurrence and absence of CHS. The other clinical or cerebral perfusion CT parameters had no significant correlation with the occurrence of CHS. CONCLUSION: Our findings suggest that patients with a prolonged dMTT of more than 3 seconds should be closely monitored for evidence of hyperperfusion after undergoing carotid stenting.

Brachial-ankle pulse wave velocity as an early indicator of left ventricular diastolic function among hypertensive subjects.

While increased arterial stiffness is a known risk of cardiovascular disease, pulse wave velocity (PWV) is a conventionally adopted index of arterial stiffness. However, the relationship between PWV and left ventricular functions are not thoroughly evaluated. This cross-sectional study investigated whether PWV measurement is an early indicator of left ventricular (LV) dysfunction. A noninvasive, volume-plethysmographic apparatus was used to determine blood pressure, electrocardiogram, heart sounds, and PWV in 42 consecutively diagnosed subjects with hypertension, and 42 sex- and age-matched nonhypertension subjects were studied. Arterial stiffness and aortic stiffness were evaluated by brachial-ankle (b-a) PWV, heart-carotid (h-c) PWV, heart-femoral (h-f) PWV, carotid-femoral (c-f) PWV, and femoral-ankle (f-a) PWV. Function of LV was estimated by tissue Doppler imaging (TDI) echocardiography. Hypertension subjects exhibited higher b-a PWV and late diastolic mitral flow velocity values than those of nonhypertensive subjects. Pearson correlation analysis revealed that LV diastolic function (Em(av)) negatively correlated with c-f PWV and b-a PWV. Multiple linear regression analysis indicated that b-a PWV was independently and negatively associated with LV diastolic function (Em(av)). Further analysis by stratified hypertensive status, the b-a PWV were independently and negatively associated with Em(av) in hypertensive subjects (p = 0.004) only. In conclusion, the b-a PWV, but not c-f PWV, h-c PWV, h-f PWV, or f-a PWV, is significantly correlated with LV diastolic function in hypertensive subjects, indicating that b-a PWV involving both central and peripheral components of arterial stiffness may be an early indicator of LV dysfunction.

Surface lattice resonance of line array of poly (glycidyl methacrylate) with CdS quantum dots for label-free biosensing.

One dimensional plasmonic grating is a kind of resonant electromagnetic wave absorber with a characteristic wavelength. This study focusses on one-dimensional plasmonic grating consisting of poly (glycidyl methacrylate) (PGMA) brushes and CdS quantum dots (CdQDs) fabrication and PGMA chains grafted on a primary substrate in a line array continued by the immobilization of biotin-modified CdQDs. PGMA brush line array (PBLA) of plasmonic grating exhibited an absorptance at 441 nm while at the same time, CdQDs immobilized with PBLA showed characteristic absorbance at 396 nm. The blue-shift from 441 nm matches the absorbance peak of biotin-modified CdQDs resulting in the enhancement of photoluminescence emission of CdQDs. With streptavidin incubation to assemble CdQDs at 50 nM, the significant decrease in grating height resulted in the red-shift of the absorbance peak to 536 nm. Due to the deviation in absorbance, the intensity of the PL emission decreased gradually with the increase in concentration of streptavidin. In addition, our results showed that streptavidin incubation altered the color reflected from the surface due to effective changes in the refractive index of the layer as well. The limit of detection of the grating for streptavidin detection was determined to be 50 nM. Thus, PBLA-CdQD has the potential to act as a highly-sensitive, label-free optical biosensor.