Application of volume-of-fluid method to analyze the viscosity effect on the spine formation of bloodstains.

In bloodstain pattern analysis, the blood droplet volume and surface impact velocity play an important role, and many related experimental studies have been carried out. If an appropriate computational fluid dynamics (CFD) model that could solve bloodstain patterns, especially spine formation bloodstain patterns, can be obtained, the blood droplet volume and impact speeds at various crime scenes can be predicted more accurately. For this purpose, Flow-3D software using the volume-of-fluid method was applied to analyze the behavior of human blood droplets during an impact event, especially focusing on the viscous effect on splashing, which forms the spine which can be used to predict the impact velocity. To obtain a non-Newtonian viscosity model of blood for a computational fluid dynamic analysis, the venous blood samples of 163 people were tested using a hemorheology instrument. Among the venous blood samples of 163 people, 37 samples for which all blood test results were in a normal range were selected for the non-Newtonian viscosity modeling. From the CFD analysis, it could be concluded that a non-Newtonian viscosity model is more appropriate than a constant viscosity model for predicting splashing that forms the spine. The gradient of the non-Newtonian model at a high shear rate has more of an effect on spine formation than that at a low shear rate. The lowest viscosity with a high velocity at the outer front of the radiating flow plays an important role in forming the splashing pattern.

Soluble syndecan-1 at diagnosis and during follow up of multiple myeloma: a single institution study.

BACKGROUND: Syndecan-1 is a heparan sulfate proteoglycan expressed on plasma cells, especially myeloma cells, and can exist in serum as soluble syndecan-1 after shedding from the cell surface. Soluble syndecan-1 has been suggested to promote myeloma cell growth and to be an independent prognostic factor for multiple myeloma. We aimed to evaluate the effect of soluble syndecan-1 levels at the time of diagnosis and during therapy on therapeutic response and prognosis for patients with multiple myeloma. METHODS: We analyzed soluble syndecan-1 levels in 28 patients with multiple myeloma and 50 normal controls, and compared its levels with Durie-Salmon stage and other markers of myeloma. In addition, we evaluated the therapeutic response and determined the 3-year survival rates of these patients. RESULTS: We observed that the median soluble syndecan-1 level in myeloma patients was higher than that in the normal controls (P <0.0001), and the soluble syndecan-1 levels in 21 (75%) patients were higher than the cut-off level (162 ng/mL). Soluble syndecan-1 levels correlated with disease stage, percentage of plasma cells in the bone marrow, ß(2) microglobulin level, serum M-component concentration, and creatinine level. The baseline levels of soluble syndecan-1 at the time of diagnosis in the patients who responded to chemotherapy were lower than those in the non-responders (P=0.04); however, the baseline level was not a significant predictor of therapeutic response. The 3-year overall survival rate of the patients with high soluble syndecan-1 levels at the time of diagnosis and 6 months after chemotherapy was lower than the corresponding survival rates of the patients with low levels of soluble syndecan-1; however, the overall survival rate was not statistically significant. CONCLUSION: The use of soluble syndecan-1 has limitations in the diagnosis of multiple myeloma. Soluble syndecan-1 levels correlate with known prognostic factors; however, we could not assess the prognostic value of high levels of soluble syndecan-1 at the time of diagnosis and after chemotherapy.

Novel swine-origin influenza A (H1N1) viral encephalitis.

The World Health Organization declared that a new strain of novel swine-origin influenza A (H1N1) virus was responsible for the pandemic infection in June 2009. We report a case of encephalitis diagnosed as the H1N1 virus infection. We describe a 17-year-old patient who had a seizure attack, diagnosed with a H1N1 virus infection via real time reverse-transcriptase polymerase chain reaction (RT-PCR). The H1N1 virus infection can be causative of the encephalitis, as with other influenza virus infections. Careful monitoring is essential for reducing complications.

The M142T mutation causes B3 phenotype: three cases and an in vitro expression study.

The B3 phenotype is the most common B subtype in Korea. The B305 allele (425 T>C, M142T) was first reported in 2 Chinese individuals; however, it has not yet been reported in the Koreans, and the impact of the M142T mutation on the expression of the B3 phenotype has also not been studied. To resolve an ABO discrepancy between a group O neonate and her group O father and A(1)B(3) mother, blood samples from these individuals and other family members were referred to our laboratory for ABO gene analysis. The B305 allele was discovered in the neonate (B305/O01), her mother (A102/ B305), and her maternal aunt (B305/O02), while her father was typed as O01/O02. Transient transfection experiments were performed in HeLa cells using the B305 allele synthesized by site-directed mutagenesis; flow cytometric analysis revealed that this transfect expressed 35.5% of the total B antigen produced by the B101 allele transfect. For comparison, Bx01 allele transfects were also created, and they expressed 11.4% of the total B antigen expressed on the surface of B101 transfects. These experiments demonstrate that the M142T (425 T>C) mutation is responsible for the B subtype phenotype produced by the B305 allele.

Origins and functional impact of copy number variation in the human genome.

Structural variations of DNA greater than 1 kilobase in size account for most bases that vary among human genomes, but are still relatively under-ascertained. Here we use tiling oligonucleotide microarrays, comprising 42 million probes, to generate a comprehensive map of 11,700 copy number variations (CNVs) greater than 443 base pairs, of which most (8,599) have been validated independently. For 4,978 of these CNVs, we generated reference genotypes from 450 individuals of European, African or East Asian ancestry. The predominant mutational mechanisms differ among CNV size classes. Retrotransposition has duplicated and inserted some coding and non-coding DNA segments randomly around the genome. Furthermore, by correlation with known trait-associated single nucleotide polymorphisms (SNPs), we identified 30 loci with CNVs that are candidates for influencing disease susceptibility. Despite this, having assessed the completeness of our map and the patterns of linkage disequilibrium between CNVs and SNPs, we conclude that, for complex traits, the heritability void left by genome-wide association studies will not be accounted for by common CNVs.

[Detection of anti-ENA and anti-dsDNA antibodies using line immunoassay in systemic autoimmune diseases].

BACKGROUND: Detection of antibodies to extractable nuclear antigens (ENAs) and dsDNA is needed for the diagnosis of and predicting prognosis in systemic autoimmune diseases. Recently introduced line immunoassay (LIA) has the advantage of detecting several autoantibodies simultaneously, and we evaluated its usefulness in the diagnosis of autoimmune diseases in comparison with enzyme-linked immunosorbent assay (ELISA). METHODS: Samples were collected from 437 patients referred by rheumatologists. FANA (fluorescent antinuclear antibody) test and LIA for the detection of 13 different autoantibodies, including 6 ENAs and dsDNA were performed. LIA-positive samples for ENA or dsDNA antibodies were further tested with ELISA. Final diagnosis was made by rheumatologists according to the diagnostic criteria. Agreement of results between LIA and ELISA was analyzed in 53 selected patients with systemic autoimmune diseases. RESULTS: The LIA detected antibodies to ENA and dsDNA in 118 and 22 patients, respectively, and ELISA detected 70.3% (83/118) and 45.5% (10/22) of LIA positive samples. Especially, 60.2% (71/118) of patients with positive ENA antibody on LIA was diagnosed as systemic autoimmune diseases. Patients having strong FANA titer and homogenous/speckled pattern showed higher prevalence of autoantibodies, but a small proportion of FANA negative patients also showed positive reactivity (LIA 10.8%, ELISA 5.2%). LIA showed a good agreement with ELISA for the anti-ENA antibodies (> or =80%), and a lower agreement for the anti-dsDNA antibody (67.9%). CONCLUSIONS: LIA detecting several autoantibodies simultaneously might replace ELISA for anti-ENA antibodies, but not for anti-dsDNA antibodies. When LIA is performed considering clinical manifestations and FANA, it could contribute to the diagnosis of systemic autoimmune disease.

[Evaluation of urine NMP22 point-of-care test for the screening of bladder cancer].

BACKGROUND: Screening of high-risk patients using bladder tumor markers can offer an advantage of early detection and saving medical costs. For these purpose many tumor markers have been developed to supplement invasive cystoscopy. Our study evaluated the NMP22 point-of-care test (NMP22 POCT), which is one of the tumor makers, comparing with the standard urine cytology for the diagnosis of bladder cancer. METHODS: From January to September 2005, 232 patients who had undergone a cystoscopy due to bladder cancer associated symptoms including hematuria and dysuria were enrolled in this study. Urine specimens were collected for NMP22 POCT and cytology. NMP22 POCT and urine cytology were compared for sensitivity and specificity. In addition, we evaluated urine stick test and microscopy to explain some false-positive results in NMP22 POCT. RESULTS: Superficial transitional cell carcinoma was diagnosed in 10 patients. The sensitivity of NMP22 test was 60% (95% confidence interval [CI], 26.2-87.8%), whereas that of cytology was 33.3% (95% CI, 7.5-70.1%); however, the difference was not significant. The specificity of NMP22 test was 69.8% (95% CI, 63.3-75.8%), compared with 99.0% (95% CI, 96.5-99.9%) for cytology (P<0.001). The presence of microscopic RBCs in urine specimen was significantly associated with the lower specificity of NMP22 POCT (P=0.02). CONCLUSIONS: NMP22 POCT was significantly less specific than urine cytology. To be useful as a bladder cancer screening test, the NMP22 test should have a higher specificity.

Microsatellite alterations in hepatocellular carcinoma and intrahepatic cholangiocarcinoma.

A series of 20 hepatocellular carcinomas and 8 intrahepatic cholangiocarcinomas was screened from the Korean population for microsatellite alterations, including a loss of heterozygosity and replication errors using nine microsatellite markers containing several genes. The microsatellite results and our previous comparative genomic hybridization results of two tumors were compared at each locus, and the correlations between these and clinicopathologic variables were examined. The most characteristic findings were found at 13q. Replication errors were prevalent at D13S160 (13q21.2 approximately q31) and D13S292(13q12). The incidence of loss of heterozygosity, however, was higher at D13S153 (13q14.1 approximately q14.3) and D13S265(13q31 approximately q32). In contrast, there were higher deletion frequencies observed in hepatocellular carcinoma (HCC) and higher amplification frequencies observed in intrahepatic cholangiocarcinoma at 13q in our previous comparative genomic hybridization (CGH) study. Higher frequencies of replication errors were observed at D16S408 (13q12 approximately q21) and D16S504(13q23 approximately q24) in the HCC. This study found that significant differences in the patterns of genetic instability of microsatellites were dependent on the chromosomal loci. It is believed that certain genes at altered CGH regions, which are relevant to the development and/or progression of these cancers, are activated by different mutation mechanisms.

Chamber-specific differentiation of Nkx2.5-positive cardiac precursor cells from murine embryonic stem cells.

Embryonic stem (ES) cells are a useful system to study cardiac differentiation in vitro. It has been difficult, however, to track the fates of chamber-specific cardiac lineages, since differentiation is induced within the embryoid body. We have established an in vitro culture system to track Nkx2.5(+) cell lineages during mouse ES cell differentiation by using green fluorescent protein (GFP) as a reporter. Nkx2.5/GFP(+) cardiomyocytes purified from embryoid bodies express sarcomeric tropomyosin and myosin heavy chain and heterogeneously express cardiac troponin I (cTnI), myosin light chain 2v (MLC2v) and atrial natriuretic peptide (ANP). After 4-week culture, GFP(+) cells exhibited electrophysiological characteristics specific to sinoatrial (SA) node, atrial, or ventricular type. Furthermore, we found that administration of 10(-7) M retinoic acid (RA) to embryoid bodies increased the percentage of MLC2v(-)ANP(+) cells; this also increased the expression of atrial-specific genes in the Nkx2.5/GFP(+) fraction, in a time- and dose-dependent fashion. These results suggest that Nkx2.5(+) lineage cells possess the potential to differentiate into various cardiomyocyte cell types and that RA can modify the differentiation potential of Nkx2.5(+) cardiomyocytes at an early stage.