Identification of a novel HLA-A*29 allele, HLA-A*29:01:09, by sequence-based typing in a Korean individual.

HLA-A*29:01:09 differs from A*29:01:01:01 by one nucleotide difference at nucleotide position 414.

Genetic association of complement component 2 polymorphism with systemic lupus erythematosus.

Complement component 2 (C2), an early member of the classical pathway, mainly participates in apoptotic cell clearance. We hypothesize that C2 polymorphism may confer genetic susceptibility to complement dysfunction in systemic lupus erythematosus (SLE). The major aim of our study was to investigate the clinical and serological associations of C2 variants in Chinese patients with SLE. The single-nucleotide polymorphism (rs2844455, G/A SNP) located in the intron region of C2 gene was genotyped by direct sequencing in 95 SLE patients and 95 matched normal control subjects. The gene expression profiles were generated by quantitative real-time polymerase chain reaction (PCR) and reverse transcription PCR. Our results showed that the AA genotype was observed more frequently in SLE patients than in normal control subjects (22.1% vs 9.5%, P < 0.05). The A allele was strongly associated with the occurrence of hair loss, photosensitivity and anti-cardiolipin antibodies; whereas, the G allele was associated with lower frequencies of these clinical presentations. Relative expression levels were significantly lower in patients with the AA genotype [median: 18.86, interquartile range (IQR) 11.36-22.43, P = 0.002] than in those with the GG genotype (35.76, IQR: 19.33-49.71). As expected, we confirmed the A allele as a risk factor for SLE development in a Chinese population, in contrast, the G allele might be a protective factor against the pathogenic autoantibody formation and cutaneous manifestations in SLE patients.

Identification of a novel HLA-A*02 allele, A*02:428, by sequence-based typing.

HLA-A*02:428 differs from A*02:06:01 by a non-synonymous mutation at codon 260 (CAT to GAT) in exon 4.

Identification of a novel HLA-C*08 allele, HLA-C*08:78, by sequence-based typing in a Korean individual.

HLA-C*08:78 differs from C*08:01:01 by a nonsynonymous mutation at codon 239 (GGA to AGA) in exon 4.

Object level HSI-LIDAR data fusion for automated detection of difficult targets.

Data fusion from disparate sensors significantly improves automated man-made target detection performance compared to that of just an individual sensor. In particular, it can solve hyperspectral imagery (HSI) detection problems pertaining to low-radiance man-made objects and objects in shadows. We present an algorithm that fuses HSI and LIDAR data for automated detection of man-made objects. LIDAR is used to define a set of potential targets based on physical dimensions, and HSI is then used to discriminate between man-made and natural objects. The discrimination technique is a novel HSI detection concept that uses an HSI detection score localization metric capable of distinguishing between wide-area score distributions inherent to natural objects and highly localized score distributions indicative of man-made targets. A typical man-made localization score was found to be around 0.5 compared to natural background typical localization scores being less than 0.1.

One-dimensional pattern of Au nanodots by ion-beam sputtering: formation and mechanism.

Highly ordered one-dimensional arrays of nanodots, or nanobeads, are fabricated by forming nanoripples and nanodots in sequence, entirely by ion-beam sputtering (IBS) of Au(001). This demonstrates the capability of IBS for the fabrication of sophisticated nanostructures via hierarchical self-assembly. The intricate nanobead pattern ideally serves to identify the governing mechanisms for the pattern formation: nonlinear effects, especially local redeposition and surface-confined transport, are essential both for the formation and the preservation of the one-dimensional order of the nanobead pattern.

A new HLA-B*15 allele, HLA-B*9587, identified by sequence-based typing in a Korean individual.

A new HLA-B*9587 showed one nucleotide difference from B*15010101 at nucleotide 127 with substitution G-->C (codon 19 GAG-->CAG) resulting in a coding change from Glu to Gln (E19Q).

Automated Measurements of Key Morphological Features of Human Embryos for IVF.

A major challenge in clinical In-Vitro Fertilization (IVF) is selecting the highest quality embryo to transfer to the patient in the hopes of achieving a pregnancy. Time-lapse microscopy provides clinicians with a wealth of information for selecting embryos. However, the resulting movies of embryos are currently analyzed manually, which is time consuming and subjective. Here, we automate feature extraction of time-lapse microscopy of human embryos with a machine-learning pipeline of five convolutional neural networks (CNNs). Our pipeline consists of (1) semantic segmentation of the regions of the embryo, (2) regression predictions of fragment severity, (3) classification of the developmental stage, and object instance segmentation of (4) cells and (5) pronuclei. Our approach greatly speeds up the measurement of quantitative, biologically relevant features that may aid in embryo selection.

Identification of a novel HLA-Cw*04 allele, HLA-Cw*040106, by sequence-based typing in the Korean population.

A new human leukocyte antigen-Cw*0401 allele showed one nucleotide difference from Cw*04010101 in exon 3 at nucleotide position 561 (codon 163 ACG-->ACT) without a coding change.

Development and clinical evaluation of a microarray for HLA-A and -DRB1 genotyping.

Microarray technology makes high-throughput genotyping possible by permitting the simultaneous analysis of large sets of genes on a small reaction slide. Human leukocyte antigen (HLA) loci showing high polymorphisms are suitable targets for microarray. In this study, we developed a microarray kit with newly designed oligonucleotide probes for the genotyping of HLA-A and -DRB1. In total, 42 probes were designed to hybridize to polymorphic sites for HLA-A and 36 for HLA-DRB1. Asymmetric polymerase chain reaction (PCR) using four primers was performed to amplify exon 2 of HLA-DRB1, whereas symmetric PCR was performed to amplify both exons 2 and 3 of HLA-A. Evaluation of performance using samples from 138 Koreans disclosed consistent microarray results with all sequence-based typing at the low-resolution level. Despite the occurrence of ambiguities in 35 HLA-A (25.4%) and 5 HLA-DRB1 (3.6%) cases, correct genotypes were assigned with high certainty by referring to allele distribution in Koreans. These data clearly indicate that our newly developed microarray kit is optimal in determining correct genotypes at the low-resolution level in Koreans.

Camellia japonica suppresses immunoglobulin E-mediated allergic response by the inhibition of Syk kinase activation in mast cells.

BACKGROUND: Novel approaches are being explored to develop new therapies for various allergic diseases. Complementary and alternative medicines are considered to be promising avenues for the development of such new therapies. OBJECTIVES: To investigate the effect of many Korean plants on the IgE-mediated allergic response in mast cells and in vivo, and its mechanism of action. MATERIALS AND METHODS: The anti-allergic activity was tested by evaluating effects on degranulation of mast cells in culture and passive cutaneous anaphylaxis (PCA) in vivo. Its mechanism of action was investigated by immunoblotting analysis, immunoprecipitation, RT-PCR, and other molecular biological approaches in mast cells. RESULTS: We screened approximately 100 natural plant extracts collected in Korea for in vitro anti-allergic activity. The leaf extract of Camellia japonica (LECJ) exhibited the most potent effect on degranulation in antigen-stimulated rodent and human mast cells. LECJ reversibly inhibited degranulation in a dose-dependent manner, with IC(50) values of approximately 50 microg/mL for the mast cells, and it also suppressed the expression and secretion of TNF-alpha and IL-4 in rat basophilic leukaemia-2H3 mast cells. In agreement with its in vitro activity, LECJ significantly inhibited mast cell-mediated PCA in an animal model. LECJ inhibited activating phosphorylation of tyrosine Y371 on Syk kinase, indicating that LECJ inhibits the activity of Src-family kinases in mast cells. In the in vitro kinase assay, LECJ directly inhibited Lyn kinase, the major Src-family kinase in the cells. It also suppressed Akt and MAP kinases, which are critical for the production of various pro-inflammatory cytokines in mast cells. In high-performance liquid chromatography analysis, quercetin-3-beta-D-glucoside and eugenol were identified as the major active components. CONCLUSION: The present results strongly suggest that the anti-allergic activity of LECJ is mediated through inhibiting degranulation and allergic cytokine secretion by inhibition of Src-family kinase in mast cells and it may be useful for the treatment of mast cell-related immediate and delayed allergic diseases.

Accuracy of Web-based recording program for in-hospital resuscitation: laboratory study.

OBJECTIVE: The purpose of this study was to assess the accuracy of a Web-based resuscitation recording program compared with the handwritten method. METHODS: A Web site was developed to record in-hospital resuscitation events and a mock resuscitation was recorded using both the Web site and handwritten method by emergency nurses. Accurate recorded events and times were compared between the two methods through the use of a video clip. Paired t tests were used to compare differences in absolute timing error, the number of omitted events out of 11 reference events and total recorded events. RESULTS: Twenty-one emergency nurses recorded simulated resuscitation events using both the handwritten and Web-based computerised recording system. The mean absolute timing errors were significantly lower using the computerised recording program (37.3 s (SD 17.1) versus 8.3 s (SD 5.3), p<0.001). The mean number of omissions for the computerised program was 1.8 (SD 0.8) compared with 1.4 (SD 1.1) for the handwritten method (p = 0.202). The mean number of total recorded events for the computerised program was 16.5 (SD 3.5) compared with 15.0 (SD 3.8) for the handwritten method (p = 0.063). CONCLUSIONS: This study suggests that a Web-based recording program decreased timing error while causing no differences in the number of recorded or omitted events in a laboratory setting.

The yeast PRP19 protein is not tightly associated with small nuclear RNAs, but appears to associate with the spliceosome after binding of U2 to the pre-mRNA and prior to formation of the functional spliceosome.

We have previously shown that the yeast PRP19 protein is associated with the spliceosome during the splicing reaction by immunoprecipitation studies with anti-PRP19 antibody. We have extended such studies by using extracts depleted of specific splicing factors to investigate the step of the spliceosome assembly process that PRP19 is involved in. PRP19 was not associated with the splicing complexes formed in U2- or U6-depleted extracts but was associated with the splicing complex formed in heat-inactivated prp2 extracts. This finding indicates that PRP19 becomes associated with the splicing complexes after or concomitant with binding of the U6 small nuclear ribonucleoprotein particle (snRNP) to the precursor RNA and before formation of the functional spliceosome. We further analyzed whether PRP19 is an integral component of snRNPs. We have constructed a strain in which an epitope of nine amino acid residues recognized by a well-characterized monoclonal antibody, 12CA5, is linked to the carboxyl terminus of the wild-type PRP19 protein. Immunoprecipitation of the splicing extracts with anti-PRP19 antibody or precipitation of the extracts prepared from the epitope-tagged strain with the 12CA5 antibody did not precipitate significant amounts of snRNAs. Addition of micrococcal nuclease-treated extracts to the PRP19-depleted extract restored its splicing activity. These results indicate that PRP19 is not tightly associated with any of the snRNAs required for the splicing reaction. No non-snRNP protein factor has been demonstrated to participate in either step of the spliceosome assembly pathway that PRP19 might be involved in. Thus, PRP19 represents a novel splicing factor.

Evaluation of a membrane bioreactor system coupled with sludge pretreatment for aerobic sludge digestion.

The effects of ozone and alkaline pretreatment of sewage sludge on the performances of membrane-coupled aerobic sludge digestion were investigated. In particular, the effects of sludge pretreatment on the solubilization, sludge biodegradation and the stability of membrane filtration were evaluated. Three sets of reactors were operated under different conditions of sludge treatment; 1) ozone treatment at 0.1 g O3 g(-1) suspended solids (SS) of the influent sludge, 2) alkaline treatment at pH 11.4-12.0, 3) no treatment. Without sludge pretreatment, 27% of suspended solids (SS) reduction was obtained at the hydraulic retention time(HRT) of 5 days. With ozone and alkaline treatment, the average SS reduction increased to 83 and 76%, respectively, at the same HRT. Membrane fouling occurred earliest with the non-treated sludge, followed by the alkali-treated sludge. With ozonated sludge, stable membrane filtration for more than 150 days was possible without chemical cleaning of the membrane. The dynamic viscosity of the mixed liquor in the reactor fed with ozonated sludge was relatively low, indicating the role of ozone treatment in controlling membrane fouling. In conclusion, sludge pretreatment followed by aerobic sludge digestion in a membrane-coupled bioreactor can achieve a very high rate of sludge reduction at a relatively short HRT.

Identification of a novel HLA-B*40 allele, HLA-B*40:332, in a Korean individual.

HLA-B*40:332 differs from B*40:01:02 by 1 nucleotide difference at nucleotide position 439.

Evaluation of a modified antimycobacterial susceptibility test using Middlebrook 7H10 agar containing 2,3-diphenyl-5-thienyl-(2)-tetrazolium chloride.

A rapid and accurate antimycobacterial susceptibility test is essential for effective treatment of tuberculosis. The aim of this study was to evaluate a modified method applying 2,3-diphenyl-5-thienyl-(2)-tetrazolium chloride (STC) to the Clinical and Laboratory Standards Institute (CLSI) guideline for susceptibility testing of Mycobacterium tuberculosis. A total of 132 clinical isolates of M. tuberculosis, forty-eight isolates showing resistance to one or more of the first-line antituberculosis drugs, and eighty-four fully susceptible isolates were collected from hospitals of a nationwide distribution from June to September 2004. The modified procedure was conducted basically according to the agar-proportion method described in the CLSI Guideline both with STC 50 mug/mL. The amount of growth in each well was recorded and graded at 2nd and 3rd weeks after inoculation. After 3 weeks of incubation, the diagnostic sensitivity and specificity for the detection of drug-resistant strains of STC-containing agar proportion methods were 100%, except ethambutol-low level resistance, of which the diagnostic sensitivity was 93.4%. After two weeks of incubation in STC-containing agar proportion methods, one hundred of the 107 strain-drug combinations have shown drug resistance, indicating the sensitivity of 93.5%. Especially, all 41 isoniazid-resistant strains and 19 of 21 rifampin-resistant strains (90.5%) could be detected after two weeks of incubation. A modification of the agar proportion method using STC resulted in a reliable and more easily interpretable data, and detected most of resistant strains a week earlier than conventional method.

The outcome of total hip replacement in obese and non-obese patients at 10- to 18-years.

We studied a consecutive series of 285 uncemented total hip replacements in 260 patients using the Taperloc femoral component and the T-Tap acetabular component. The outcome of every hip was determined in both living and deceased patients. A complete clinical and radiological follow-up was obtained for 209 hips in 188 living patients, followed for a mean of 14.5 years (10 to 18.9). They were divided into two groups, obese and non-obese, as determined by their body mass index. There were 100 total hip replacements in 89 patients in the obese cohort (body mass index > or = 30 kg/m(2)), and 109 in 99 non-obese (body mass index < 30 kg/m(2)) patients. A subgroup analysis of 31 patients of normal weight (body mass index 20 kg/m(2) to 25 kg/m(2)) (33 hips) and 26 morbidly obese patients (body mass index > or = 35 kg/m(2)) (30 hips) was also carried out. In the obese group five femoral components (5%) were revised and one (1%) was loose by radiological criteria. Femoral cortical osteolysis was seen in eight hips (8%). The acetabular component was revised in 57 hips (57%) and a further 17 (17%) were loose. The mean Harris hip score improved from 52 (30 to 66) pre-operatively to 89 (49 to 100) at final follow-up. Peri-operative complications occurred in seven patients (7%). In the non-obese group six (6%) femoral components were revised and one (1%) was loose. Femoral cortical osteolysis occurred in six hips (6%). The acetabular component was revised in 72 hips (66%) and a further 18 (17%) were loose. The mean Harris hip score increased from 53 (25 to 73) prior to surgery to 89 (53 to 100) at the time of each patient's final follow-up radiograph. No statistically significant difference was identified between the obese and non-obese patients with regards to clinical and radiological outcome or complications. The subgroup analysis of patients of normal weight and those who were morbidly obese showed no statistically significant difference in the rate of revision of either component. Our findings suggest there is no evidence to support withholding total hip replacement from obese patients with arthritic hips on the grounds that their outcome will be less satisfactory than those who are not obese.

Long-term results of uncemented total hip arthroplasty with the Taperloc femoral component in patients with Dorr type C proximal femoral morphology.

AIMS: To investigate the longevity of uncemented fixation of a femoral component in total hip arthroplasty (THA) in patients with Dorr type C proximal femoral morphology. PATIENTS AND METHODS: A total of 350 consecutive uncemented THA in 320 patients were performed between 1983 and 1987, by a single surgeon using the Taperloc femoral component. The 63 patients (68 hips) with Dorr type C proximal femoral morphology were the focus of this review. The mean age of the patients was 69 years (24 to 88) and mean follow-up was 16.6 years (ten to 29). Survival analysis included eight patients (eight hips) who died without undergoing revision surgery prior to obtaining ten years follow-up. All 55 surviving patients (60 hips) were available for clinical assessment and radiographic review. As a comparator group, the survival and implant fixation in the remaining 282 THAs (257 patients) with Dorr type A and B morphology were evaluated. The mean age of these patients was 52 years (20 to 82). RESULTS: In the Dorr C patient group the mean Harris hip score improved from 51 points (21 to 69 points) pre-operatively to 89 (74 to 100) at final follow-up. No femoral component was loose by radiological criteria and osteolysis was only identified around two stems (3.3%). There was one revision (1.6%) of a well-fixed femoral component for sepsis at 11 years. The survival of the Taperloc femoral component at 20 years with revision for any reason as the endpoint was 98% (95% confidence interval; 87 to 99). A total of ten (3.5%) of the Dorr A and B patient group of 282 THAs required revision surgery. Only one (0.4%) for aseptic loosening. A total of two hips (1%) were loose by radiographic criteria and osteolysis occurred around 12 hips (4%). CONCLUSION: This study demonstrates that excellent fixation can be achieved using the Taperloc stem in patients with Dorr type A and B, and Dorr type C bone. TAKE HOME MESSAGE: The Taperloc stem demonstrated equivalent results in Dorr type A and B and Dorr type C bone. Cite this article: Bone Joint J 2016;98-B:595-600.

Evidence of a new metabolic pathway of 5-fluorouracil in Escherichia coli from in vivo 19F-NMR spectroscopy.

Two distinct metabolic pathways of 5-fluorouracil are proposed in Escherichia coli. The first metabolic pathway is a reductive degradation with the formation of dihydrofluorouracil as the first metabolite. The second metabolic pathway is shown to be a hydroxylating degradation, possibly with the formation of 5-hydro-6-hydroxy-5-fluorouracil as the first metabolite. The metabolites of both pathways undergo subsequent hydrolytic degradation with fluoride ion as the common final product. The chemical structures of these metabolites were partially identified by 19F-NMR. The results show a close resemblance between these two metabolic pathways with in vivo pyrimidine biodegradation. The reductive degradation has been proposed by several laboratories, whereas the hydroxy degradation has not been reported before. Both the reductive and hydroxy pathways are demonstrated in this report, to be independent reactions.