Ultra-high mass multimer analysis of protein-1a capping domains by a silicon nanomembrane detector.

Conventional time of flight ion detectors are based on secondary electron multipliers encountering a significant loss in detection efficiency, sensitivity and resolution with protein mass above 50kDa. In this work we employ a silicon nanomembrane detector in a Matrix-Assisted Laser Desorption/Ionization coupled to time of flight (MALDI-TOF) mass spectrometer. The operating principle relies on phonon-assisted field emission with excellent performance in the high mass range from 0.001-2MDa. In addition to the analysis of standard proteins the nanomembrane detector (NMD) has the potential for the detection and structural investigation of complex macromolecular assemblies through non-covalent interactions. In order to investigate this hypothesis, the N-terminal capping/methyltransferase domain (CAP) of the Brome Mosaic Virus (BMV) 1a replication protein by MALDI-TOF-NMD is analyzed. The signals detected at the high m/z-ratios of 912.6/982.7 (×103) and 1333.3 (×103) could be modified species of CAP-tricta/tetractamer and the octadecamer. For the first time, the NMD is applied to detect biologically complex macromolecular protein assemblies. Hence, this technology overcomes the limitations of conventional TOF-detectors and increases the analytical range of MALDI-TOF. This technology will be a future alternative for the structural analysis of intact virus capsids that will complement other MS-based techniques such as native mass spectrometry.

Clinical benefit of nomogram for predicting positive resection margins in breast conserving surgery.

PURPOSE: Previously, we reported a nomogram for the prediction of positive resection margin (RM) after breast conserving surgery (BCS). This study was conducted to evaluate the clinical usefulness of the nomogram. METHODS: Prospective patients who underwent operations using the nomogram between July 2012 and August 2013 (nomogram group; N = 260) were compared with past control patients who underwent operations between July 2010 and October 2011 and underwent frozen section biopsy (FSB) without use of the nomogram (N = 266). In the nomogram group, an intraoperative assessment of RM using FSB was only performed when the nomogram score was higher than predefined cut-off (>80). In addition, we conducted retrospective analysis of additional 181 patients who received BCS in another institute (Kyoto University Hospital). These patients did not undergo FSBs for RMs. RESULTS: Of 260 patients, 161 (61.9%) presented low nomogram scores and avoided FSB. The surgical decision to use the nomogram did not significantly increase reoperation rate due to positive RM compared with the control FSB group (4.6% vs. 3.8%, p = 0.47). The surgery time was significantly reduced by 18.1% (mean 14.7 min) in nomogram group (p < 0.001). Of 99 nomogram high-score patients, 14 presented with positive RM on FSB and 11 of them avoided reoperation. In the Kyoto cohort, the reoperation rate was significantly lower in low-score patients than in high-score patients (2.7% vs. 11.4%, p < 0.001). CONCLUSIONS: We showed that our nomogram is useful to reduce FSBs without increasing reoperation rate for surgeons who perform routine FSBs. For most surgeons, it can give useful information about the possibility of tumor-positive RMs.

Simultaneous determination of water-soluble whitening ingredients and adenosine in different cosmetic formulations by high-performance liquid chromatography coupled with photodiode array detection.

OBJECTIVE: The Korean Cosmetic Act regulates the use of functional cosmetics) by the law. Four functional cosmetic groups, whitening, anti-wrinkle, UV protection and combination of whitening and anti-wrinkle, were categorized according to the Korean Cosmetic Act and Functional Cosmetics Codex. In this study, high-performance liquid chromatography (HPLC) coupled with photodiode array detection (DAD) was employed for the simultaneous detection of arbutin (and its decomposition product, hydroquinone), niacinamide, ascorbyl glucoside, ethyl ascorbyl ether and adenosine in functional cosmetic products such as creams, emulsions and lotions. METHODS: Separation by HPLC-DAD was conducted using a C18 column with a gradient elution of 5 mm KH2PO4 buffer (containing 0.1% phosphoric acid) and methanol (containing 0.1% phosphoric acid). The wavelengths for the detection of arbutin, hydroquinone, niacinamide, adenosine, ascorbyl glucoside and ethyl ascorbyl ether were 283, 289, 261, 257, 238 and 245 nm, respectively. RESULTS: This method exhibited good linearity (R(2) ≥ 0.999), precision (expressed as relative standard deviation (RSD) < 2%) and mean recoveries (89.42-104.89%). The results obtained by monitoring 100 market samples showed that the detected levels of the tested materials are within the acceptable authorized concentration. CONCLUSION: The method developed herein is simple and can be used for market survey and quality control of functional cosmetics.

Simultaneous determination of arbutin and its decomposed product hydroquinone in whitening creams using high-performance liquid chromatography with photodiode array detection: Effect of temperature and pH on decomposition.

OBJECTIVE: Arbutin is an effective agent for the treatment of melanin disorders. Arbutin may be converted to hydroquinone under conditions of high temperature, ultraviolet (UV) radiation and dilute acid. The aim of the current study was to develop an analytical method to determine the levels of arbutin and hydroquinone in whitening cosmetic products using high-performance liquid chromatography with photodiode array detection (HPLC-DAD). In addition, we investigated the effects of high temperature and pH on the decomposition of arbutin. METHODS: Samples extracted using two-step sonications were separated on a C18 column using a gradient mobile phase consisting of water and methanol. A 60-mm (40 µL) DAD cell was used to enhance the sensitivity of hydroquinone determination. Thermal decomposition of arbutin was evaluated at temperatures ranging from 60 to 120°C for 1-36 h. RESULTS: The method showed good linearity (R(2) ≥ 0.9997), precision (relative standard deviation, RSD < 5%) and acceptable extraction recovery (90-102.6%). The limits of quantitation for arbutin and hydroquinone were 0.0085 and 0.0119 µg mL(-1) , respectively. One sample of 21 cosmetic products tested contained arbutin at a concentration 1.61 g 100 g(-1) cream and 0.12 g 100 g(-1) cream of hydroquinone. Arbutin (327.18 ppm) decomposed after 6 h at 120°C and produced 10.73 ppm of hydroquinone. CONCLUSION: The developed method is simple to detect both arbutin and hydroquinone simultaneously in cosmetic products, at an adequate level of sensitivity. Notably, temperature and pH did not influence the decomposition of arbutin to hydroquinone in a 2% arbutin cream.

Temperature dependence of the electronic structure of two-dimensional Na gas on the Si(111)-7 × 7 surface.

The temperature dependence of the irreversible phase transition from a two-dimensional gas to an ordered zero-dimensional solid on the Si(111)-7 × 7 surface was studied using photoemission spectroscopy. With increasing Na coverage, the two-dimensional Na gas, which is a state of highly mobile Na atoms, undergoes a phase transition into ordered zero-dimensional magic nanoclusters at room temperature. The critical Na coverage of the phase transition was found to increase with reduced temperature. This was used to develop a gas-solid phase diagram of Na atoms on the Si(111)-7 × 7 surface as a function of Na coverage and sample temperature based on the electronic structure. The temperature dependence of the phase transition can be ascribed to the suppression of the thermal energy that is required to overcome the energetic barrier between the two-dimensional gas and the zero-dimensional solid at low temperature, where three different hopping mechanisms are related to the phase transition.

Diagnostic performance of multidetector CT for acute cholangitis: evaluation of a CT scoring method.

OBJECTIVE: The aim of this study was to retrospectively assess the diagnostic performance of multidetector CT (MDCT) for the diagnosis of acute cholangitis using a new scoring method. METHODS: Of 80 patients with suspected biliary disease who underwent biphasic CT and endoscopic retrograde cholangiography, 39 were diagnosed as having acute cholangitis (Group 1) and 41 patients were classified as suspected biliary disease (Group 2). 100 age-matched patients without evidence of biliary disease were selected randomly as a control group (Group 3). Each patient's axial scan was scored by two independent radiologists for the extent of transient hepatic attenuation difference, the presence of biliary dilatation and identification of a biliary obstructive lesion. The difference in the scores among the three groups was evaluated and the optimal cut-off score for the diagnosis of acute cholangitis was determined. Interobserver agreement was also evaluated. RESULTS: The total scores (mean ± standard deviation) for Groups 1, 2 and 3 were 7.0 ± 2.0, 4.4 ± 2.4 and 0.9 ± 1.2, respectively, for Reviewer 1 and 7.2 ± 2.7 and 0.7 ± 1.1, respectively, for Reviewer 2. Significant differences were found for the subscores and the total scores among the three groups (p < 0.001). Using a cut-off score of ≥ 5, the sensitivity and specificity for diagnosing acute cholangitis were 84.6% and 83.7%, respectively, for Reviewer 1 and 89.7% and 83.7%, respectively, for Reviewer 2. Agreement for the subscores between readers was good to excellent (κ = 0.74-0.86). CONCLUSION: Based on dynamic MDCT and the described CT scoring method, the diagnosis of acute choangitis can be made with high sensitivity and specificity.

Microtubule associated protein 2 and choline acetyltransferase immunoreactivity in the lumbar spinal cord of young adult and aged dogs.

German Shepherds are a good model for research about aging and neurological disorders such as lumbosacral spinal canal stenosis. We compared neurons, glia and cholinergic neurons in the ventral horn of the lumbar spinal cord (L(3)) between adult (1-2 years old) and aged (10-12 years old) groups. Any pathological findings were not found by hematoxylin and eosin staining and neurological examination, and the number of NeuN (a marker for neurons)-positive neurons were similar in both groups. Microtubule-associated protein 2 (MAP2) immunoreactive dendrites in the aged dog were decreased without any change in ß-tubulin protein level. Glial fibrillary acidic protein (a marker for astrocytes) and ionized calcium-binding adapter molecule 1 (a marker for microglia) immunoreactivity were not significantly changed in both groups. The number of ChAT immunoreactive neurons was decreased; however, its protein level was not significantly changed in the aged group. These results suggest that numbers of ventral horn neurons are not changed, but cholinergic neurons may change in aged dogs compared to adult dogs.

Immunoreactivity and protein levels of olfactory marker protein and tyrosine hydroxylase are not changed in the dog main olfactory bulb during normal ageing.

The immunoreactivity and protein expression of olfactory marker protein (OMP) and tyrosine hydroxylase (TH) in the main olfactory bulb (MOB) of the dog during normal ageing was investigated. OMP immunolabelling was observed only in nerve bundles of the olfactory nerve (ONL) and glomerular layers (GL) and there was no OMP expression within cell bodies of any layer. TH immunolabelling was detected in all layers of the MOB except for the ONL. Most of the neurons expressing TH were distributed in the juxtaglomerular region and had a morphological appearance consistent with periglomerular, external tufted or superficial short axon cells. Dendrites of TH-immunoreactive neurons were closely apposed to OMP-immunoreactive nerve bundles within the glomeruli. There was no significant age-related loss of OMP and TH immunoreactivity and protein concentrations of these molecules were consistent in dogs of different ages. These results suggest that olfactory signal transduction to the GL via axons of olfactory receptor neurons remains unchanged during ageing in the dog.

Pharmacokinetics and milk distribution characteristics of orbifloxacin following intravenous and intramuscular injection in lactating ewes.

The purpose of the current investigation is to elucidate the pharmacokinetic profiles of orbifloxacin (OBFX) in lactating ewes (n = 6) following intravenous (i.v.) and intramuscular (i.m.) administrations of 2.5 mg/kg W. In a crossover study, frequent blood, milk, and urine samples were drawn for up to 48 h after the end of administration, and were then assayed to determine their respective drug concentrations through microbiological assay using Klebsiella pneumoniae as the test micro-organism. Plasma pharmacokinetic parameters were derived from plasma concentration-time data using a compartmental and noncompartmental analysis, and validated a relatively rapid elimination from the blood compartment, with a slope of the terminal phase of 0.21 +/- 0.02 and 0.19 +/- 0.06 per hour and a half-life of 3.16 +/- 0.43 and 3.84 +/- 0.59 h, for i.v. and i.m. dosing, respectively. OBFX was widely distributed with a volume of distribution V((d(ss))) of 1.31 +/- 0.12 L/kg, as suggested by the low percentage of protein binding (22.5%). The systemic body clearance (Cl(B)) was 0.32 +/- 0.12 L/h x kg. Following i.m. administration, the maximum plasma concentration (C(max)) of 1.53 +/- 0.34 microg/mL was reached at t(max) 1.25 +/- 0.21 h. The drug was completely absorbed after i.m. administration, with a bioavailability of 114.63 +/- 11.39%. The kinetic milk AUC(milk)/AUC(plasma) ratio indicated a wide penetration of orbifloxacin from the bloodstream to the mammary gland. OBFX urine concentrations were higher than the concurrent plasma concentrations, and were detected up to 30 h postinjection by both routes. Taken together, these findings indicate that systemic administration of orbifloxacin could be efficacious against susceptible mammary and urinary pathogens in lactating ewes.

Multiple gastrointestinal stromal tumours presenting as a haemoperitoneum in a patient with Type 1 neurofibromatosis: MDCT findings.

We report multidetector row CT images of multiple gastrointestinal tumours in a patient with Type 1 neurofibromatosis who presented with haemoperitoneum. The use of CT demonstrated multiple, variably sized, ovoid, relatively well-enhancing masses in the intramural or subserosal location of the jejunum. The largest mass that showed a haemorrhagic necrosis caused haemoperitoneum. These imaging features are unique and differ from previous imaging features that have been reported in the radiological literature.

Duplication of the spleen with a short pancreas.

The most common form of splenic anomaly with a concurrent short pancreas is polysplenia, which has been described in various studies in the radiological literature. However, splenic duplication has never been reported. We report here a case of splenic duplication associated with a short pancreas and pre-duodenal portal vein. This extremely rare case of splenic anomaly shows unique multidetector CT findings that are distinguishable from a splenic lobulation or cleft.

Characterization of the pharmacokinetic disposition of levofloxacin in stallions after intravenous and intramuscular administration.

The target of the present study was to investigate the plasma disposition kinetics of levofloxacin in stallions (n = 6) following a single intravenous (i.v.) bolus or intramuscular (i.m.) injection at a dose rate of 4 mg/kg bwt, using a two-phase crossover design with 15 days as an interval period. Plasma samples were collected at appropriate times during a 48-h administration interval, and were analyzed using a microbiological assay method. The plasma levofloxacin disposition was best fitted to a two-compartment open model after i.v. dosing. The half-lives of distribution and elimination were 0.21 +/- 0.13 and 2.58 +/- 0.51 h, respectively. The volume of distribution at steady-state was 0.81 +/- 0.26 L/kg, the total body clearance (Cl(tot)) was 0.21 +/- 0.18 L/h/kg, and the areas under the concentration-time curves (AUCs) were 18.79 +/- 4.57 microg.h/mL. Following i.m. administration, the mean t(1/2el) and AUC values were 2.94 +/- 0.78 h and 17.21 +/- 4.36 microg.h/mL. The bioavailability was high (91.76% +/- 12.68%), with a peak plasma mean concentration (C(max)) of 2.85 +/- 0.89 microg/mL attained at 1.56 +/- 0.71 h (T(max)). The in vitro protein binding percentage was 27.84%. Calculation of efficacy predictors showed that levofloxacin might have a good therapeutic profile against Gram-negative and Gram-positive bacteria, with an MIC

Dissipation behavior of lufenuron, benzoylphenylurea insecticide, in/on Chinese cabbage applied by foliar spraying under greenhouse conditions.

Chinese cabbage has long been consumed as a staple food by the Koreans in various forms of fresh, salted, or fermented as kimchi. To fulfill the off-season demand for this crop, it has become a common practice to be cultivated under greenhouse conditions. Since pesticide residues in/on leafy vegetables have strongly concerned with food safety in the Korean society, the changes of lufenuron residues, in/on Chinese cabbage applied by foliar spraying under greenhouse conditions was investigated. Lufenuron 5% emulsifiable concentrate (EC) was sprayed with diluted solution of recommended and double doses to the crop. The shoots of the cabbage were harvested immediately after spraying, and sequentially the harvests were conveyed to analyze the residual amounts. The deposited level of the analyte in/on Chinese cabbage under greenhouse conditions seemed to be difficult to produce the crop with 0.2 ppm of maximum residue limit (MRL) of the Korea Food and Drug Administration (KFDA).

Progressive collapse of PMMA-augmented vertebra: a report of three cases.

Vertebroplasty using polymethylmethacrylate (PMMA) for augmentation is accepted as a safe and effective treatment for vertebral compression fracture. However, various complications related to PMMA vertebroplasty have recently been reported. We experienced three cases with progressive collapse of PMMA-augmented vertebra. Collapse progressed after augmentation in cases where PMMA conglomerated without contiguous bone interdigitation. A high viscosity of the PMMA preparation and vertebral body cavitory lesion may play a role in progressive vertebral collapse. To avoid this complication, bone cement should be injected sufficiently and permeate to contiguous bone to create strong support and anchorage.

Surgical management of spinal cord haemangioblastoma.

BACKGROUND: The surgical management of spinal cord haemangioblastomas is distinct from that of other benign spinal cord tumours and optimal surgical strategy is still being determined because of the rarity of the condition. The aim of this study is to investigate factors that affect the outcome of surgical management. PATIENTS AND METHODS: We retrospectively analysed 24 operations for symptomatic spinal cord haemangioblastomas in 20 patients. Clinical features and surgical results were investigated by medical record review, telephone interviews, angiographic images, and magnetic resonance images (MRI). The mean follow-up period was 5.6 years (range 6 months to 13.6 years). RESULTS: Patients with cystic components showed pre-operative motor weakness and sensory change more commonly than those without cystic components. Post-operative function scale had a positive correlation with pre-operative function (R(2) = 0.727; p < 0.001) and no correlation with the extent of the surgery. All subtotally removed tumours recurred, whereas totally removed tumours recurred in only 3 patients. CONCLUSION: The cystic component of spinal cord haemangioblastomas is responsible for symptom generation and is helpful for dissecting tumours. Post-operative functional status is determined by pre-operative functional status. Total removal is feasible by using the correct surgical technique and is recommended to prevent recurrence.

Prevalence of hepatitis E virus and sapovirus in post-weaning pigs and identification of their genetic diversity.

Hepatitis E virus (HEV) and sapovirus (SaV) induce acute hepatitis and gastroenteritis, respectively, in humans. As pigs have been recognized as an important reservoir for these viruses, we evaluated the infection rates of both viruses using fecal samples collected from post-weaning pigs via RT-PCR methods. In the five swine farms assessed in this study, 3 farms were found to be HEV-positive and 4 farms were SaV-positive. The overall prevalence of HEV and SaV in the pigs was 17.0 and 23.1%, respectively. Phylogenetic tree analysis revealed that the isolated swine HEVs belonged to genotype 3 and the porcine SaVs belonged to genogroup III. This study proved that both HEV and SaV are prevailing in post-weaning pigs in Korea.

Co-prescribing of proton pump inhibitors among chronic users of NSAIDs in the UK.

OBJECTIVES: Co-prescribing of proton pump inhibitors (PPIs) with non-selective NSAIDs (nsNSAIDs) is recommended in patients at risk of gastrointestinal (GI) events. This study estimated usage of PPI co-therapy among chronic nsNSAID users and determined factors associated with concurrent nsNSAID-PPI use. METHODS: The retrospective study was based on the Intercontinental Marketing Services (IMS) Health UK MediPlus database and included subjects > or = 40 yrs of age who received their first oral nsNSAID prescription between July and December 2002 and who had > or = 60 days of nsNSAID supply during the following year. Days with nsNSAID-PPI overlap were calculated and logistic regression was used to identify factors associated with nsNSAID-PPI overlap. A generalized linear model was used to assess the degree of association of GI risk factors with the nsNSAID-PPI overlap ratio among PPI users. RESULTS: Of 16,344 patients included, 1586 received at least one PPI prescription. Among PPI users, PPIs were available on approximately 50% of the days with nsNSAID therapy. After multivariate adjustment, age > or = 65 yrs, history of any hospitalization and co-prescriptions for anti-coagulants or oral corticosteroids increased the odds of any nsNSAID-PPI overlap by 21-68%. Prior gastroprotective agent (GPA) use increased the odds of any PPI use during follow-up 16-fold and nsNSAID-PPI overlap 19-fold. Among PPI users, patients with prior use of any GPA had a 2.46 times higher nsNSAID-PPI overlap ratio. CONCLUSIONS: PPI utilization correlates poorly with nsNSAID use in the UK. GI safety of nsNSAID-PPI co-therapy observed in controlled trials may therefore not be achieved in clinical practice.

Pharmacokinetics and mammary residual depletion of erythromycin in healthy lactating ewes.

The aim of this investigation was to examine the pharmacokinetics and mammary excretion of erythromycin administered to lactating ewes (n = 6) by the intravenous (i.v.), intramuscular (i.m.) and subcutaneous (s.c.) routes at a dosage of 10 mg/kg. Blood and milk samples were collected at pre-determined times, and a microbiological assay method was used to measure erythromycin concentrations in serum and milk. The concentration-time data were analysed by compartmental and non-compartmental kinetic methods. The serum concentration-time data of erythromycin were fit to a two-compartment model after i.v. administration and a one-compartment model with first-order absorption after i.m. and s.c. administration. The elimination half-life (t(1/2beta)) was 4.502 +/- 1.487 h after i.v. administration, 4.874 +/- 0.296 h after i.m. administration and 6.536 +/- 0.151 h after s.c. administration. The clearance value (Cl tot) after i.v. dosing was 1.292 +/- 0.121 l/h/kg. After i.m. and s.c. administration, observed peak erthyromycin concentrations (Cmax) of 0.918 +/- 0.092 microg/ml and 0.787 +/- 0.010 microg/ml were achieved at 0.75 and 1.0 h (Tmax) respectively. The bioavailability obtained after i.m. and s.c. administration was 91.178 +/- 10.232% and 104.573 +/- 9.028% respectively. Erythromycin penetration from blood to milk was quick for all the routes of administration, and the high AUC milk/AUC serum (1.186, 1.057 and 1.108) and Cmax-milk/Cmax-serum ratios reached following i.v., i.m. and s.c. administration, respectively, indicated an extensive penetration of erythromycin into the milk.