Pediococcus pentosaceus-Fermented Cordyceps militaris Inhibits Inflammatory Reactions and Alleviates Contact Dermatitis.

Cordyceps militaris is a medicinal mushroom used to treat immune-related diseases in East Asia. We investigated the anti-inflammatory effect of the extract of C. militaris grown on germinated Rhynchosia nulubilis (GRC) fermented with Pediococcus pentosaceus ON89A isolated from onion (GRC-ON89A) in vivo as well as in vitro. The anti-inflammatory effect of GRC-ON89A was investigated in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. The total polyphenol content (TPC) and total flavonoid content (TFC) in the GRC-ON89A ethanol extract were significantly increased compared to that in GRC. GRC-ON89A hexane fraction (GRC-ON89A-Hex) inhibited the release of nitric oxide (NO) compared to that of the LPS-treated control without cytotoxicity in LPS-stimulated RAW 264.7 macrophages. GRC-ON89A-Hex decreased the inducible NO synthase (iNOS), cyclooxygenase 2 (COX2), and tumor necrosis factor (TNF)-α mRNA expression in LPS-stimulated RAW 264.7 macrophages. In addition, pre-treatment with GRC-ON89A-Hex significantly inhibited LPS-stimulated phosphorylation of mitogen-activated protein kinases (MAPKs) and nuclear factor (NF)-κB. To induce allergic contact dermatitis (ACD), 1-fluoro-2, 4-dinitrofluorobenzene (DNFB) was applied to the surface of the right ears of C57BL/6N mice. GRC-ON89A reduced the ear swelling and thickness in DNFB-induced ACD mice. This study demonstrates the potential usefulness of GRC-ON89A as an anti-inflammatory dietary supplement or drug.

Sensing Properties of ZnO Nanoparticles for Detection of 2-Chloroethyl Ethyl Sulfide as a Mustard Simulant.

In this work, we present the fabrication and characterization of a 2-chloroethyl ethyl sulfide (2-CEES) gas sensor based on ZnO nanoparticles (NPs) synthesized by a hydrothermal method. We confirmed that synthesized ZnO NPs adopt a polycrystalline phase. Partially aggregated ZnO-NPs revealed spherical or ellipsoidal nanocrystalline particles in a size range of 30-50 nm, as observed by field-emission scanning electron microscopy (FE-SEM). The maximum response of the ZnO NPs was 15 at 1 ppm 2-CEES concentration, and a low detection limit of 0.4 ppm was observed at an optimal operating temperature of 250 °C. The lowest response time was 6 s in 20 ppm at 250 °C. The linearity response with correlation coefficient (R2) was 0.9887 at 2-CEE concentrations of 0.4-1 ppm at the operating temperature of 250 °C. The enhanced sensing performance and a decrease in the operating temperature were attributed to a high specific surface area and more active sites in the ZnO NPs after exposure to 2-CEES.

Evaluation of Surface Cleaning Procedures in Terms of Gas Sensing Properties of Spray-Deposited CNT Film: Thermal- and O2 Plasma Treatments.

The effect of cleaning the surface of single-walled carbon nanotube (SWNT) networks by thermal and the O2 plasma treatments is presented in terms of NH3 gas sensing characteristics. The goal of this work is to determine the relationship between the physicochemical properties of the cleaned surface (including the chemical composition, crystal structure, hydrophilicity, and impurity content) and the sensitivity of the SWNT network films to NH3 gas. The SWNT networks are spray-deposited on pre-patterned Pt electrodes, and are further functionalized by heating on a programmable hot plate or by O2 plasma treatment in a laboratory-prepared plasma chamber. Cyclic voltammetry was employed to semi-quantitatively evaluate each surface state of various plasma-treated SWNT-based electrodes. The results show that O2 plasma treatment can more effectively modify the SWNT network surface than thermal cleaning, and can provide a better conductive network surface due to the larger number of carbonyl/carboxyl groups, enabling a faster electron transfer rate, even though both the thermal cleaning and the O2 plasma cleaning methods can eliminate the organic solvent residues from the network surface. The NH3 sensors based on the O2 plasma-treated SWNT network exhibit higher sensitivity, shorter response time, and better recovery of the initial resistance than those prepared employing the thermally-cleaned SWNT networks.

Paternal Somatic Mosaicism of a Novel Frameshift Mutation in ELANE Causing Severe Congenital Neutropenia.

Severe congenital neutropenia (SCN) is a bone marrow failure disease with an autosomal dominant inheritance from mutations in ELANE. Here, we report a 7-week-old Korean male with SCN. His elder sister died from pneumonia at 2 years. Direct sequencing of ELANE in the proband identified a heterozygous novel frameshift mutation: c.658delC (p.Arg220Glyfs20*). Family study involving his asymptomatic parents with normal cell counts revealed that his father had the same mutation, but at a lower burden than expected in a typical heterozygous state. Further molecular investigation demonstrated somatic mosaicism with ~18% mutant alleles. We concluded the proband inherited the mutation from his somatic mosaic father.

Label-free electrochemical detection of the human adenovirus 40/41 fiber gene.

A nanoporous silicon-based label-free DNA biosensor was fabricated to monitor rapidly enteric adenovirus types 40 and 41, a leading cause of viral gastroenteritis in children. Nanoporous silicon (NPS) was formed by an anodic etching process in a mixture solution containing hydrofluoric acid and ethanol. The polypyrrole (PPy) film was directly electropolymerized on The NPS substrate. Twenty-five base pairs of probe DNA (pDNA), derived from the fiber gene, was electrochemically doped on the PPy-coated NPS substrate. The conductivity change due to the immobilized pDNA and hybridized target DNA (tDNA) was expressed as an arbitrary factor, γ, which is a normalized numerical term used for the selective quantification of the tDNA. γ was inversely proportional to the concentration of complementary tDNA, but independent of the non-complementary tDNA. The sensitivity slope for detecting tDNAc was -1.54 µM(-1), based on the factor γ in the range of 0.4 to 1.0 µM of tDNA. The surface roughness was characterized using atomic force microscopy.

Electrochemical serotonin monitoring of poly(ethylenedioxythiophene):poly(sodium 4-styrenesulfonate)-modified fluorine-doped tin oxide by predeposition of self-assembled 4-pyridylporphyrin.

A 5,10,15,20-tetrakis(4-pyridyl)-21H,23H-porphyrin (TPyP)-modified self-assembled functional layer was prepared on a fluorine-doped tin oxide (FTO) substrate. We employed a bifunctional molecule, 3-iodopropionate (3IP), to covalently bind TPyP to the FTO substrate. The 3IP-monolayered FTO and the TPyP-3IP-bilayered FTO electrodes were characterized by cyclic voltammetry, electrochemical impedance spectroscopy, and Fourier transform-infrared spectroscopy. Compared to conventional electropolymerized poly(ethylenedioxythiophene):poly(sodium 4-styrenesulfonate) (PEDOT:PSS) film on bare FTO, the PEDOT:PSS film on the TPyP-3IP-bilayered FTO showed better sensitivity and selectivity in monitoring serotonin in the presence of high concentrations of interfering agents such as ascorbic acid, urea, D-(+)-glucose, epinephrine, and L-3,4-dihydroxyphenylalanine. Both PEDOT:PSS films on the bare FTO and the TPyP-3IP-bilayered FTO showed electrocatalytic effects in serotonin detection, and only the TPyP-3IP-based PEDOT:PSS film acted as a pH resistant buffer layer in the selective detection of serotonin.

Dispersion of nanodiamond on chemical mechanical polishing performance for Ge1Sb6Te3 film.

This study describes the effect of surfactant concentration on the chemical mechanical polishing process of Ge1Sb6Te3 film using nanodiamond-based slurry. Aggregated diamond nanoparticles were dispersed in a slurry solution containing anionic poly(sodium 4-styrene sulfonate) using milling system. The zeta-potential, particle size and transmission electron microscopy image of the dispersed nanodiamond particles were analyzed for slurries having varying surfactant concentrations to identify the effect of the surfactant concentration on the milling process. The cationic nanodiamond particles were covered with the anionic poly(sodium 4-styrene sulfonate) polymer, and the polymer acted as a dispersion agent on account of the electrostatic repulsion. By increasing the surfactant concentration in the milling process, the average particle size of the nanodiamond particle decreased until the concentration reached 0.9 wt%. In addition, the surface roughness and material removal rate of the Ge1Sb6Te3 film in the polishing process strongly-depended on the surfactant concentration. Both surface roughness and material removal rate decreased with an increase in the surfactant concentration. Excess poly(sodium 4-styrene sulfonate) acted as a passivation layer, resulting in a decrease in the surface roughness and material removal rate of the Ge1Sb6Te3 film.

A Novel Frameshift Mutation of the USH2A Gene in a Korean Patient with Usher Syndrome Type II.

Usher syndrome type II (USH2) is the most common form of Usher syndrome, characterized by moderate to severe hearing impairment and progressive visual loss due to retinitis pigmentosa. It has been shown that mutations in the USH2A gene are responsible for USH2. The authors herein describe a 34-year-old Korean woman with the typical clinical manifestation of USH2; she had bilateral hearing disturbance and progressive visual deterioration, without vestibular dysfunction. Molecular genetic study of the USH2A gene revealed a novel frameshift mutation (c.2310delA; Glu771LysfsX17). She was heterozygous for this mutation, and no other mutation was found in USH2A, suggesting the possibility of an intronic or large genomic rearrangement mutation. To the best of our knowledge, this is the first report of a genetically confirmed case of USH2 in Korea. More investigations are needed to delineate genotype-phenotype correlations and ethnicity-specific genetic background of Usher syndrome.

Dopamine sensor based on a boron-doped diamond electrode modified with a polyaniline/Au nanocomposites in the presence of ascorbic acid.

A selective dopamine (DA) sensor was developed using gold nanoparticles (Au-NPs) dispersed overoxidized-polyaniline (PANI(ox)) based on a boron-doped diamond (BDD) thin-film electrode. The concentration of the DA was determined using voltammetry as a non-enzymatic sensor. BDD thin film has a high signal-to-noise ratio, a long-term stability, a high sensitivity, and a good reproducibility. PANI nanocomposites were directly synthesized on the BDD electrode and overoxidized using 0.5 M H(2)SO(4) solution. The overoxidized PANI film enhances selectivity and sensitivity toward DA. The Au-NPs were dispersed on the PANI nanocomposite by electrochemical deposition. The nanometer-sized Au-NPs favor the sensing of DA in the presence of ascorbic acid (AA). The combination of the PANI with the Au-NPs and the BDD electrode can create synergetic effects for the performance of the biosensor, such as a fast response time, a lower detection limit, a wider linear range, enhanced selectivity, and higher sensitivity for the determination of DA.

Diagnostic utility of a multiplex RT-PCR assay in detecting fusion transcripts from recurrent genetic abnormalities of acute leukemia by WHO 2008 classification.

Fusion transcripts (FT) from chromosomal rearrangements are key culprits in acute leukemia, with genotype-phenotype correlations including prognostic implications. Here, we report our experience of a commercially available platform utilizing multiplex reverse-transcriptase polymerase chain reaction (RT-PCR), HemaVision, in 309 consecutive patients with acute leukemia. A total of 108 patients (35%) were diagnosed as having acute leukemia with recurrent genetic abnormalities by the World Health Organization 2008 classification. The multiplex RT-PCR platform, detected 12 different FT in 92 (85.2%; 92/108), with a 99% concordance rate with conventional cytogenetics/fluorescence in situ hybridization. Additional information obtained from the multiplex RT-PCR assay included transcript heterogeneity and novel splice variants of FT. In addition, the RT-PCR assay targeting specific FT could be used for monitoring minimal residual disease. HemaVision is a robust diagnostic platform in detecting FT in routine clinical laboratories both at initial diagnosis and for disease monitoring.

Estimation of carrier frequencies of six autosomal-recessive Mendelian disorders in the Korean population.

Although many studies have been performed to identify mutations in Korean patients with various autosomal-recessive Mendelian disorders (AR-MDs), little is known about the carrier frequencies of AR-MDs in the Korean population. Twenty common mutations from six AR-MDs, including Wilson disease (WD), non-syndromic hearing loss (NSHL), glycogen storage disease type Ia (GSD Ia), phenylketonuria (PKU), congenital hypothyroidism (CH), and congenital lipoid adrenal hyperplasia (CLAH) were selected to screen for based on previous studies. A total of 3057 Koreans were genotyped by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry followed by confirmation using the Sanger sequencing. We found 201 and 8 carriers with either one or two mutations in different genes, respectively, yielding a total carrier frequency of 1 in 15 (6.7%). Of the six AR-MDs, NSHL has the highest carrier frequency followed by WD, CH, CLAH, GSD Ia, and PKU. As carrier screening tests are becoming prevalent and the number of mutations known and tested is rising, a priori data on the carrier frequencies in different ethnic groups is mandatory to plan a population screening program and to estimate its efficiency. In light of this, the present results can be used as a basis to establish a screening policy for common AR-MRs in the Korean population.

Heterogeneous lengths of copy number mutations in human coagulopathy revealed by genome-wide high-density SNP array.

BACKGROUND: The recent advent of genome-wide molecular platforms has facilitated our understanding of the human genome and disease, particularly copy number aberrations. We performed genome-wide single nucleotide polymorphism-array in hereditary coagulopathy to delineate the extent of copy number mutations and to assess its diagnostic utility. DESIGN AND METHODS: The study subjects were 17 patients with hereditary coagulopathy from copy number mutations in coagulation genes detected by multiple ligation-dependent probe amplification. Eleven had hemophilia (7 hemophilia A and 4 hemophilia B) and 6 had thrombophilia (4 protein S deficiency and 2 antithrombin deficiency). Single nucleotide polymorphism-array experiments were performed using Affymetrix Genome-Wide Human SNP arrays 6.0. RESULTS: Copy number mutations were identified by single nucleotide polymorphism-array in 9 patients, which ranged in length from 51 Kb to 6,288 Kb harboring 2 to ~160 genes. Single nucleotide polymorphism-array showed a neutral copy number status in 8 patients including 7 with either a single-exon copy number mutation or duplication mutations of PROS1. CONCLUSIONS: This study revealed unexpectedly heterogeneous lengths of copy number mutations underlying human coagulopathy. Single nucleotide polymorphism-array had limitations in detecting copy number mutations involving a single exon or those of a gene with homologous sequences such as a pseudogene.

Fabrication of Pt nanoparticles-decorated CVD diamond electrode for biosensor applications.

An electrochemical biosensor was developed using boron-doped diamond (BDD) as an electrode material. To enhance the electrical performance of the electrode, the BDD electrode was decorated with Pt-nanoparticles (Pt-NPs) by electrochemical deposition. Their morphology according to the applied potentials for the synthesis of Pt-NPs was characterized by SEM. To identify the performance of the electrode modified with Pt-NPs, glucose detection was used as a sample sensing process, and the results were compared with those of a gold electrode and a bare BDD electrode. The electrochemical characteristics of the modified electrode were examined by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The BDD electrode with the Pt-NPs showed higher sensitivity and a lower detection limit than the Au electrode and BDD electrode. The proposed biosensor based on the Pt-NPs decorated BDD electrode showed high sensitivity, a low detection limit, fast direct electron transfer and good stability.

Underestimation of recipient DNA in bone marrow by post-transplant chimerism analyses using DNA extracted from EDTA-collected aspirate samples in pediatric acute myeloid leukemia.

GOALS: Chimerism analysis (CA) is essential for post-transplant surveillance. DNA from bone marrow (BM) aspirates drawn into EDTA specimen tubes (EDTA-BM) is widely used for CA. Since EDTA-BM is subject to peripheral dilution, however, DNA from aspirate particle smears (PS-BM) might better represent BM chimerism status. In this regard, we evaluated the influence of BM sources on CA. PROCEDURES: Study subjects were consecutive pediatric AML patients who had experienced relapse after allogeneic stem cell transplantation with the interval between CA before relapse (pre-relapse) and at relapse <6 months. We compared chimerism status at the 2 time-points by fluorescence PCR on STR markers using EDTA-BM vs PS-BM. RESULTS: Eight patients were eligible for this study. The recipient DNA (%R) from EDTA-BM was 0% at pre-relapse in all except 2 with 1.6% and 1.3%, while %R using PS-BM revealed mixed chimerism in all 8, %R ranging 1.6-13.2% (median 3.75%). The %R from EDTA-BM was 0.9-79.3% at relapse (29.15%), while %R from PS-BM was 3.8-86.6% (60.15%). The difference of %R (Δ%R), %R[PS-BM]-% R[EDTA-BM], was median 3.15% (1.6-13.2%) at pre-relapse and median 12.1% (2.1-60.7%) at relapse. CONCLUSIONS: Our study showed CA using EDTA-BM significantly underestimated %R. Our observation might have a ramification to other quantitative workup in hematologic malignancy.

CKIT mutation in therapy-related acute myeloid leukemia with MLLT3/MLL chimeric transcript from t(9;11)(p22;q23).

Gain-of-function mutations of the CKIT gene have been reported to specifically occur in core-binding factor (CBF) acute myeloid leukemia (AML) with a poor prognostic implication. Here we report a case of therapy-related AML with t(9;11)(p22;q23) who had CKIT mutation. A 48-year-old woman with breast cancer received partial mastectomy followed by 6 cycles of adjuvant chemotherapy and radiation therapy. At 28 months from the diagnosis of breast cancer, she was diagnosed as having AML with blasts 81% in bone marrow. Cytogenetic analysis revealed t(9;11)(p22;q23), and FISH showed 96.5% of MLL break-apart signals. RT-PCR study revealed MLL(11q23)/MLLT3(9p22) chimeric transcript. FLT3-ITD and NPM1 mutations were both negative. Unexpectedly, mutation analyses for CKIT identified D816Y mutation. The patient received induction chemotherapy and achieved complete remission at 1 month. To the best of our knowledge, this is the first report on CKIT mutation in therapy-related AML with MLL rearrangement.

Molecular characterization of female hemophilia A by multiplex ligation-dependent probe amplification analysis and X-chromosome inactivation study.

Hemophilia A is an X-linked recessive bleeding disorder caused by mutations in the F8 gene. Hemophilia A typically occurs in male individuals, but female patients with hemophilia A have rarely been reported. Here we describe molecular characteristics of three unrelated female patients with severe hemophilia A of Korean descent. Patient 1 was a 5-year-old girl and was found to be compound heterozygous for intron 22 inversion inherited from her father with hemophilia A and a large deletion mutation from her mother. The large deletion detected by multiplex ligation-dependent probe amplification involved the whole F8 gene. Patient 2 was a 30-year-old woman and was heterozygous for small duplication mutation in exon 14 (c.3275dupA; p.Asn1092LysfsX26). Patient 3 was a 16-year-old girl and was heterozygous for intron 22 inversion. All three patients showed nonrandom X-chromosome inactivation status. The results underscore the need for a meticulous search for another mutation in the maternally derived X-chromosome such as large-dosage mutations.

Direct deposition of patterned nanocrystalline CVD diamond using an electrostatic self-assembly method with nanodiamond particles.

Micron-sized and precise patterns of nanocrystalline CVD diamond were fabricated successfully on substrates using dispersed nanodiamond particles, charge connection by electrostatic self-assembly, and photolithography processes. Nanodiamond particles which had been dispersed using an attritional milling system were attached electrostatically on substrates as nuclei for diamond growth. In this milling process, poly sodium 4-styrene sulfonate (PSS) was added as an anionic dispersion agent to produce the PSS/nanodiamond conjugates. Ultra dispersed nanodiamond particles with a ζ-potential and average particle size of - 60.5 mV and ∼ 15 nm, respectively, were obtained after this milling process. These PSS/nanodiamond conjugates were attached electrostatically to a cationic polyethyleneimine (PEI) coated surface on to which a photoresist had been patterned in an aqueous solution of the PSS/nanodiamond conjugated suspension. A selectively seeded area was formed successfully using the above process. A hot filament chemical vapor deposition system was used to synthesize the nanocrystalline CVD diamond on the seeded area. Micron-sized, thin and precise nanocrystalline CVD diamond patterns with a high nucleation density (3.8 ± 0.4 × 10(11) cm(-2)) and smooth surface were consequently fabricated.

The common NF-κB essential modulator (NEMO) gene rearrangement in Korean patients with incontinentia pigmenti.

Incontinentia pigmenti (IP) is a rare X-linked dominant disorder characterized by highly variable abnormalities of the skin, eyes and central nervous system. A mutation of the nuclear factor-κB essential modulator (NEMO) located at Xq28 is believed to play a role in pathogenesis and the mutation occurs mostly in female patients due to fatal consequence of the mutation in males in utero. This study was designed to identify the common NEMO rearrangement in four Korean patients with IP. Deletion of exons 4 to 10 in the NEMO, the most common mutation in IP patients, was detected in all of the patients by the use of long-range PCR analysis. This method enabled us to discriminate between NEMO and pseudogene rearrangements. Furthermore, all of the patients showed skewed XCI patterns, indicating pathogenicity of IP was due to cells carrying the mutant X chromosome. This is the first report of genetically confirmed cases of IP in Korea.

[Determination of urinary iodine concentration by inductively coupled plasma-mass spectrometry in thyroid cancer patients on low-iodine diet].

BACKGROUND: Thyroid cancer patients should be on low-iodine diet (LID) before radioactive iodine therapy (RAIT) to maximize the effect of RAIT. Urinary iodine excretion is the most accurate marker of very recent dietary iodine intake. We developed and evaluated the analytical performance of inductively coupled plasma-mass spectrometry (ICP-MS) to determine urinary iodine concentration. METHODS: We evaluated the linearity, precision, accuracy, and lower limit of quantification (LLOQ) of an ICP-MS method (Agilent 7500ce) to determine urinary iodine concentration in accordance with the Food and Drug Administration (FDA) guidelines for bioanalytical method validation. This method was used to determine and compare the iodine concentration in random urine samples of 120 thyroid cancer patients on LID for 1 week and 80 healthy adults on normal diet. RESULTS: Our ICP-MS method showed good linearity (1.0-1,913 microg/L; R(2)>0.999). Both intra-day and inter-day precision CV were within 20% for the LLOQ (1 microg/L) and within 15% for the other concentrations. Accuracy was 110-120% for the LLOQ and 95-115% for the other concentrations. The median concentration of iodine in random urine samples from thyroid cancer patients on LID (38.7 microg/L) was significantly lower than that of healthy subjects (238.8 microg/L) (P<0.0001). CONCLUSIONS: Urinary iodine analysis by ICP-MS showed good linearity, precision, accuracy, wide measuring range of detection, and lower LLOQ. This method will be very useful to evaluate the status of dietary iodine intake and the appropriateness of LID in thyroid cancer patients, thereby maximizing the effect of RAIT.

Clinical usefulness of fluorescence in situ hybridization for diagnosis and surveillance of bladder cancer.

We evaluated the performance of the fluorescence in situ hybridization (FISH) assay in comparison with that of urinary cytology for the detection of bladder urothelial carcinoma in routine clinical practice. Voided urine samples from 602 patients with hematuria were analyzed. The bladder cancer group consisted of 95 patients who had biopsy-proven bladder cancer, and the control group consisted of 507 patients without bladder cancer. We found a significant difference between the overall sensitivities of FISH and cytology (60% vs. 28.4%, respectively; P < 0.0001). The overall specificity was 99% with cytology and 94.9% with FISH, although this difference was not statistically significant. The mean values for all four probes in the true-positive group were higher than those in the false-positive group. The difference in the mean values between the two groups was significant only for the CEP3 and CEP17 probes. Furthermore, the severity of the genetic alterations detected by FISH showed a positive correlation with both tumor invasiveness (stage Ta --> T1, T2) and histological grade (G1, G2 --> G3). Together, these findings suggest that FISH can be a useful diagnostic and surveillance tool for patients who are suspected of having new or recurrent bladder cancer.