[Auto-analysis of immunohistochemical findings for breast cancer using specified software and virtual microscopy].

Currently, the therapeutic strategy for a breast cancer patient is designed according to their histopathological, immunohistochemical and molecular findings. These findings are obtained through the collected efforts of many individual pathologists or medical technologists (MTs) and are, thus, limited by intra-observer error and potentially subjective decision making. Twenty five breast cancer specimens collected between November 2009 and February 2010 were examined for immunohistochemical expressions of estrogen receptor (ER), progesterone receptor (PgR), HER2, Ki-67, Topoisomerase II alpha (Topo IIalpha). Fifty one cancer specimens collected November 2009 and June 2010 were examined for human epidermal growth factor 2 (HER2). Immunohistochemical staining was performed using auto-stainers (Ventana) and the results were stored digitally after examination by virtual microscopy (Hamamatsu Photonics). Data analysis was performed with the Genie/Aperio software package on a desk-top computer. For all the antibodies used expect for HER2, concordant results were obtained in 100% of 24 ER positive cases. Ki-67 index (r=0.96) and Topo IIalpha index (r=0.95) also showed a significant correlation (p<0.001). For HER2, all four specimens with Hercep-score 2 by ocular observation but auto-analysis score 1 revealed no HER2 gene amplification. Well-organized auto-analysis is more likely to result in an objective observation and to provide a means by which to standardize the methods for immunohistochemical detection of breast cancer.

Missense mutation of Abcg5 in stroke-prone spontaneously hypertensive rats does not influence lymphatic sitosterol absorption regardless of the dose: comparison with Wistar rats.

The lymphatic recovery of radiolabeled sitosterol administered in various amounts to the stomach was almost the same between stroke-prone spontaneously hypertensive rats (SHRSPs), a strain having a missense mutation in ATP binding cassette transporter g5 (Abcg5), and Wistar rats, a normal strain. The results suggest that the mutation of Abcg5 in SHRSPs, compared with Wistar rats, did not influence the ability for intestinal sitosterol absorption regardless of the dose.

Missense mutation in Abcg5 in SHRSP rats does not accelerate intestinal absorption of plant sterols: comparison with Wistar rats.

Stroke-prone spontaneously hypertensive rats (SHRSP) deposit plant sterols in their bodies and have a mutation in ATP binding cassette transporter G5 (Abcg5). Lymphatic recovery rates of campesterol and sitosterol in SHRSP rats were comparable to those in Wistar rats, a strain that does not deposit plant sterols in the body and has no mutation in Abcg5. Higher absorption of stigmasterol and sitostanol was observed in SHRSP rats than in Wistar rats, but the differences between SHRSP and Wistar rats were quite small, because the absorbed amounts of these two sterols were much lower than those of campesterol and sitosterol. The in situ uptake of (3)H-sitosterol and (14)C-cholesterol solubilized in the bile salt micelle into intestinal mucosa was comparable between SHRSP and Wistar rats. These observations suggest that a mutation in Abcg5 does not greatly influence intestinal absorption of plant sterols in SHRSP rats, at least in comparison with Wistar rats.

Hydrolysis of micellar phosphatidylcholine accelerates cholesterol absorption in rats and Caco-2 cells.

Lymphatic recovery of cholesterol infused into the duodenum as bile salt micelles containing phosphatidylcholine (PC) was accelerated by the co-administration of phospholipase A2 in bile and pancreatic juice diverted rats. Previously we observed that cholesterol esterase, which has the ability to hydrolyze PC, caused the same effect under a similar experimental condition (Ikeda et al., Biochim. Biophys. Acta, 1571, 34-44 (2002)). Accelerated cholesterol absorption was also observed when a part of micellar PC was replaced by lysophosphatidylcholine (LysoPC) and oleic acid. Phospholipase A2 facilitated the incorporation of micellar cholesterol into Caco-2 cells in a dose-dependent manner. There was a highly negative correlation between the incorporation of cholesterol into Caco-2 cells and the content of micellar PC remaining in the culture medium. The release of cholesterol as a monomer from bile salt micelles was enhanced when a part of micellar PC was replaced with LysoPC and oleic acid. These results strongly suggest that the release of monomer cholesterol from bile salt micelles is accelerated by hydrolysis of PC in bile salt micelles and hence that cholesterol absorption is enhanced.

Circular dichroism of amino acid films in UV-VUV region.

Circular dichroism (CD) spectra of sublimated films of alanine and alanine dipeptide were measured in UV-VUV region. When a film has some linear anisotropy, the apparent CD signal is known to be strongly distributed by linear anisotropy. In order to obtain the true CD spectra, we examined this disturbance for L-alanine films. We measured linear dichroism spectra and CD spectra as a function of the sample rotation angle, turning the sample over. On the basis of these measurements, we concluded that the linear anisotropy contribution to CD was sufficiently small for carefully prepared films. In addition, we revealed that it is essential for accurate measurement to prepare thin film of which thickness is smaller than 100 nm for the case of alanine films. CD of amino acid films was compared with that of aqueous solution. Sign of CD signal of solid films was observed to be the same for aspartic acid, but it was reversed from that of aqueous solution for alanine and alanine dipeptide.