RESUMO
Primary cerebellar glioblastoma is a rare disease that accounts for 0.4-3.4% of glioblastoma multiforme(GBM)cases. The clinicopathological characteristics and prognosis of primary cerebellar GBM are not well understood due to its rarity and the lack of an established treatment strategy. To elucidate the prognostic factors and dissemination pattern, we retrospectively assessed four cases of cerebellar GBM that we treated between 2003 and 2013. All cases involved men, and the age range was 53 to 76 years(median 69.5 years);each patient underwent surgical removal and received adjuvant chemotherapy or radiotherapy. Every cerebellar GBM patient developed intrathecal dissemination at every stage of cerebellar GBM. Two patients had spinal metastases with tumor recurrence, and no patient had brain stem invasion. IDH1 mutation and MGMT expression were both negative in three cases. The median overall survival of cerebellar GBM patients was 13.8 years, and the median progression-free survival was 5.5 years, which is similar to that reported in previous reports-and similar in terms of results-for supratentorial GBM treated at the same time at our institution. In conclusion, the prognosis of cerebellar GBM appears to be similar to that of supratentorial GBM;however, the pattern of tumor progression, such as intrathecal dissemination, is different. Craniospinal irradiation on cerebellar GBM should be carefully considered with frequent follow-up by whole spine survey using MRI.
Assuntos
Neoplasias Encefálicas , Neoplasias Cerebelares , Glioblastoma , Idoso , Humanos , Masculino , Pessoa de Meia-Idade , Recidiva Local de Neoplasia , Prognóstico , Estudos RetrospectivosRESUMO
When inert gas containing water molecules flows into a metal pipe, the water molecules cannot exit instantaneously from the outlet of the pipe but are captured at adsorption sites on the inner surface of the pipe until most of the sites are occupied. A theoretical model and a subsequent experiment in this article show that the delay time depends on the amount of moisture level; the higher the moisture-level, the shorter the delay time. Based on the result, we propose a new method and its implementation to the validation of a standard moisture generation to be used in the field measurement such as in factories and pipe lines.
RESUMO
A VeraCode-allele-specific primer extension (ASPE) method was applied to the detection and genotyping of human papillomavirus (HPV)-DNA. Oligonucleotide primers containing HPV-type-specific L1 sequences were annealed to HPV-DNA amplified by PGMY-PCR, followed by ASPE to label the DNA with biotinylated nucleotides. The labeled DNA was captured by VeraCode beads through hybridization, stained with a streptavidin-conjugated fluorophore, and detected by an Illumina BeadXpress® reader. By using this system, 16 clinically important HPV types (HPV6, 11, 16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58, 59, 66 and 68) were correctly genotyped in a multiplex format. The VeraCode-ASPE genotyping of clinical DNA samples yielded identical results with those obtained by validated PGMY-reverse blot hybridization assay, providing a new platform for high-throughput genotyping required for HPV epidemiological surveys.
Assuntos
Alphapapillomavirus/isolamento & purificação , Reação em Cadeia da Polimerase Multiplex/métodos , Infecções por Papillomavirus/virologia , Alelos , Alphapapillomavirus/classificação , Alphapapillomavirus/genética , Primers do DNA/genética , Feminino , Genótipo , Humanos , Reação em Cadeia da Polimerase Multiplex/instrumentação , Infecções por Papillomavirus/diagnósticoRESUMO
Molecularly targeted agents for cancer therapy are recognized as being effective and are gaining in popularity. However, the efficacy of the agents depends on the status of the targeted molecule such as the number of molecules expressed, activity, and mutation. Therefore, the use of companion diagnostics for investigating the status of the targeted molecule prior to therapy is highly important. We developed a simple and cost-effective somatic mutation detection method called the fluorescence resonance energy transfer-based preferential homoduplex formation assay (FRET-PHFA). By using double-stranded labeled DNA and fluorescence measurement with thermal control, this method provides higher reproducibility, easier handling, less risk for contamination, shorter assay time (only â¼15min), and less cost compared with conventional PHFA. Here we report the evaluation of FRET-PHFA on the detection of multiallelic KRAS mutations in codons 12 and 13 compared with the TheraScreen clinical diagnostics kit. We found that FRET-PHFA detected KRAS mutations (1.25-50%) from all cell line DNA titration samples.
Assuntos
Análise Mutacional de DNA/métodos , Transferência Ressonante de Energia de Fluorescência/métodos , Animais , Linhagem Celular Tumoral , Humanos , Camundongos , Mutação , Reação em Cadeia da Polimerase , Proteínas ras/genéticaRESUMO
The measurement and control of trace moisture, where the water concentration is lower than 1 ppmv [-76.2 °C for the frost point (°CFP)], are essential for improving the yield rate of semiconductor devices and for ensuring their reliability. A ball surface acoustic wave (SAW) sensor with a sol-gel silica coating exhibited useful characteristics for a trace moisture analyzer (TMA) when the temperature drift of the delay time output was precisely compensated using two-frequency measurement (TFM), where the temperature-compensated relative delay time change (RDTC) was obtained by subtracting the RDTC at the fundamental frequency from that at the third harmonic frequency on an identical propagation path. However, the cost of the measurement circuit was a problem. In this study, a burst waveform undersampling (BUS) circuit based on the theory of undersampling measurement was developed as a practical means. The BUS circuit was useful for precise temperature compensation of the RDTC, and the ball SAW TMA was prototyped by calibrating the RDTC using a TMA based on cavity ring-down spectroscopy (CRDS), which is the most reliable method for trace moisture measurement. The ball SAW TMA outputted a similar concentration to that obtained by the CRDS TMA, and its response time at a set concentration in N2 with a flow rate of 1 l/min was about half that of the CRDS TMA, suggesting that moisture of -80 °CFP was measured within only 1 min. The detection limit at a signal-to-noise ratio of 3 was estimated to be 0.05 ppbv, comparable with that of the CRDS TMA. From these results, it was demonstrated that a practical ball SAW TMA can be realized using the developed BUS circuit.
RESUMO
A thin beam of wave usually diverges due to diffraction, which is a limitation of any device using such waves. However, a surface acoustic wave (SAW) on a sphere with an appropriate aperture does not diverge but is naturally collimated, realizing ultramultiple roundtrips along an equator of the sphere. This effect is caused by the balance between diffraction and focusing on a spherical surface, and it enables realization of high-performance ball SAW sensors. The advantage of ball SAW is most fully appreciated when applied to a very thin sensitive film for which the multiple-roundtrip enhances the sensitivity, but the attenuation loss is not very large. It is exemplified in a hydrogen gas sensor that realizes a wide sensing range of 10 ppm to 100% for the first time, and realizes relatively fast response time of 20 s without heating the sensitive film.
RESUMO
We analyzed the acoustic waves propagating in a sphere to establish a useful guideline for the design of NDE apparatus and ball surface acoustic wave (SAW) device exploiting the diffraction-free propagation of SAW on a sphere. First, we calculated the laser-generated acoustic displacements both under ablation condition and under thermoelastic condition and verified experimentally the validity of the calculation. Next, the acoustic waves excited by out-of-plane stress and those excited by in-plane stress were compared. The results showed that when the out-of-plane stress was applied, the relative amplitudes of the bulk waves to that of the SAW were larger and the number of bulk waves was larger than that when the in-plane stress was applied, while the SAW had similar waveforms in each case. The ratio of the relative amplitude of the bulk waves for the out-of-plane stress and the in-plane stress was 3.1:1 at phi(1)=90 degrees and 1.67:1 at phi(1)=0 degrees. The large amplitude for the out-of-plane stress can be explained by wide directivities of bulk waves. Consequently, we found that it is necessary for ball SAW device to select a piezoelectric material and form of interdigital transducer so that the in-plane stress becomes dominant.
RESUMO
BACKGROUND: KRAS, BRAF and PIK3CA mutations are frequently observed in colorectal cancer (CRC). In particular, KRAS mutations are strong predictors for clinical outcomes of EGFR-targeted treatments such as cetuximab and panitumumab in metastatic colorectal cancer (mCRC). For mutation analysis, the current methods are time-consuming, and not readily available to all oncologists and pathologists. We have developed a novel, simple, sensitive and fully automated molecular diagnostic system (AMDS) for point of care testing (POCT). Here we report the results of a comparison study between AMDS and direct sequencing (DS) in the detection of KRAS, BRAF and PI3KCA somatic mutations. METHODOLOGY/PRINCIPAL FINDING: DNA was extracted from a slice of either frozen (nâ=â89) or formalin-fixed and paraffin-embedded (FFPE) CRC tissue (nâ=â70), and then used for mutation analysis by AMDS and DS. All mutations (nâ=â41 among frozen and 27 among FFPE samples) detected by DS were also successfully (100%) detected by the AMDS. However, 8 frozen and 6 FFPE samples detected as wild-type in the DS analysis were shown as mutants in the AMDS analysis. By cloning-sequencing assays, these discordant samples were confirmed as true mutants. One sample had simultaneous "hot spot" mutations of KRAS and PIK3CA, and cloning assay comfirmed that E542K and E545K were not on the same allele. Genotyping call rates for DS were 100.0% (89/89) and 74.3% (52/70) in frozen and FFPE samples, respectively, for the first attempt; whereas that of AMDS was 100.0% for both sample sets. For automated DNA extraction and mutation detection by AMDS, frozen tissues (nâ=â41) were successfully detected all mutations within 70 minutes. CONCLUSIONS/SIGNIFICANCE: AMDS has superior sensitivity and accuracy over DS, and is much easier to execute than conventional labor intensive manual mutation analysis. AMDS has great potential for POCT equipment for mutation analysis.