Improved methods using the reverse transcriptase polymerase chain reaction to detect tumour cells

Reverse transcriptase polymerase chain reaction (RT-PCR) is increasingly used to detect small numbers of circulating tumour cells, though the clinical benefit remains controversial. The largest single contributing factor to the controversy of its value is the different approaches to sample processing. The aim of this study was to compare the sensitivity and reproducibility of RT-PCR for the detection of tumour cells after four commonly used different methods of sample processing. Using RT-PCR, one tumour cell spiked in 2 ml of whole blood was detected after analysis of separated mononuclear cell RNA, whole blood total or poly-A+RNA. No false positives were identified with any method. However, the reproducibility of tumour cell detection was reduced after isolation of the mononuclear cell fraction. Only analysis of poly-A+RNA had a sensitivity of 100% in all the cell spiking experiments. In patient blood samples, analysis of poly-A+RNA increased the number of blood samples positive for tyrosine hydroxylase (TH) mRNA compared with those positive after analysis of total RNA. This may reflect high levels of cDNA reducing the efficiency of the PCR. Isolation of poly-A+RNA increases the sensitivity and reproducibility of tumour cell detection in peripheral blood. © 1999 Cancer Research Campaig

High sensitivity GEM experiment on double beta decay of 76-Ge

The GEM project is designed for the next generation 2 beta decay experiments with 76-Ge. One ton of ''naked'' HP Ge detectors (natural at the first GEM-I phase and enriched in 76-Ge to 86% at the second GEM-II stage) are operating in super-high purity liquid nitrogen contained in the Cu vacuum cryostat (sphere with diameter 5 m). The latest is placed in the water shield. Monte Carlo simulation evidently shows that sensitivity of the experiment (in terms of the T1/2 limit for neutrinoless 2 beta decay) is 10^27 yr with natural HP Ge crystals and 10^28 yr with enriched ones. These bounds corresponds to the restrictions on the neutrino mass less than 0.05 eV and 0.015 eV with natural and enriched detectors, respectively. Besides, the GEM-I set up could advance the current best limits on the existence of neutralinos - as dark matter candidates - by three order of magnitudes, and at the same time would be able to identify unambiguously the dark matter signal by detection of its seasonal modulation.Comment: LaTeX, 20 pages, 4 figure

The trail of my studies on glycoproteins from enterokinase to tumor markers

This review describes the results of the author’s studies on glycoproteins which have been carried out for more than 50 years. Starting from the elucidation of basic structures of glycoproteins, i.e. the structure of the linkage between an amino acid and a sugar and the occurrence of the β-mannosidic linkage as the common structure of glycoproteins, the author became interested in the cell membrane glycoproteins focused on the comparison of cancer cells versus normal cells. These studies were then extended to the establishment of sugar-directed and cancer-associated monoclonal antibodies. Some of the monoclonal antibodies are useful for cancer diagnosis

Susceptibility of hamsters to clostridium difficile isolates of differing toxinotype

Clostridium difficile is the most commonly associated cause of antibiotic associated disease (AAD), which caused ~21,000 cases of AAD in 2011 in the U.K. alone. The golden Syrian hamster model of CDI is an acute model displaying many of the clinical features of C. difficile disease. Using this model we characterised three clinical strains of C. difficile, all differing in toxinotype; CD1342 (PaLoc negative), M68 (toxinotype VIII) and BI-7 (toxinotype III). The naturally occurring non-toxic strain colonised all hamsters within 1-day post challenge (d.p.c.) with high-levels of spores being shed in the faeces of animals that appeared well throughout the entire experiment. However, some changes including increased neutrophil influx and unclotted red blood cells were observed at early time points despite the fact that the known C. difficile toxins (TcdA, TcdB and CDT) are absent from the genome. In contrast, hamsters challenged with strain M68 resulted in a 45% mortality rate, with those that survived challenge remaining highly colonised. It is currently unclear why some hamsters survive infection, as bacterial and toxin levels and histology scores were similar to those culled at a similar time-point. Hamsters challenged with strain BI-7 resulted in a rapid fatal infection in 100% of the hamsters approximately 26 hr post challenge. Severe caecal pathology, including transmural neutrophil infiltrates and extensive submucosal damage correlated with high levels of toxin measured in gut filtrates ex vivo. These data describes the infection kinetics and disease outcomes of 3 clinical C. difficile isolates differing in toxin carriage and provides additional insights to the role of each toxin in disease progression

Improved Measurement of the K+ to pi+ nu nubar Branching Ratio

An additional event near the upper kinematic limit for K+ to pi+ nu nubar has been observed by Experiment E949 at Brookhaven National Laboratory. Combining previously reported and new data, the branching ratio is B(K+ to pi+ nu nubar)= 1.47 (+1.30, - 0.89) x 10-10 based on three events observed in the pion momentum region 211<P<229 MeV/c. At the measured central value of the branching ratio, the additional event had a signal-to-background ratio of 0.9

Species traits and geomorphic setting as drivers of global soil carbon stocks in seagrass meadows

Our knowledge of the factors that can influence the stock of organic carbon (OC) that is stored in the soil of seagrass meadows is evolving, and several causal effects have been used to explain the variation of stocks observed at local to national scales. To gain a global-scale appreciation of the drivers that cause variation in soil OC stocks, we compiled data on published species-specific traits and OC stocks from monospecific and mixed meadows at multiple geomorphological settings. Species identity was recognized as an influential driver of soil OC stocks, despite their large intraspecific variation. The most important seagrass species traits associated with OC stocks were the number of leaves per seagrass shoot, belowground biomass, leaf lifespan, aboveground biomass, leaf lignin, leaf breaking force and leaf OC plus the coastal geomorphology of the area, particularly for lagoon environments. A revised estimate of the global average soil OC stock to 20 cm depth of 15.4 Mg C ha−1 is lower than previously reported. The largest stocks were still recorded in Mediterranean seagrass meadows. Our results specifically identify Posidonia oceanica from the Mediterranean and, more generally, large and persistent species as key in providing climate regulation services, and as priority species for conservation for this specific ecosystem service

Further Evidence for the Decay K+ to pi+ neutrino-antineutrino

Additional evidence for the rare kaon decay K+ to pi+ neutrino-antineutrino has been found in a new data set with comparable sensitivity to the previously reported result. One new event was observed in the pion momentum region examined, 211<P<229 MeV/c, bringing the total for the combined data set to two. Including all data taken, the backgrounds were estimated to contribute 0.15 pm 0.05 events. The branching ratio is B=1.57^{+1.75}_{-0.82} 10^{-10}.Comment: 10 pages, 2 figure

Search for the decay K+ to pi+ gamma gamma in the pi+ momentum region P>213 MeV/c

We have searched for the K+ to pi+ gamma gamma decay in the kinematic region with pi+ momentum close to the end point. No events were observed, and the 90% confidence-level upper limit on the partial branching ratio was obtained, B(K+ to pi+ gamma gamma, P>213 MeV/c) < 8.3 x 10-9 under the assumption of chiral perturbation theory including next-to-leading order ``unitarity'' corrections. The same data were used to determine an upper limit on the K+ to pi+ gamma branching ratio of 2.3 x 10-9 at the 90% confidence level.Comment: 15 pages, 3 figures; no change in the results, accepted for publication in Physics Letters