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Notes: Identification of apple clonal rootstocks based on visual morphological characteristics is often difficult because of the genetic similarity of many rootstocks and the restriction of observable characteristics at the nursery stage. In this study, a total of 66 rootstock clones representing a sub-sample of a collection of apple rootstocks from different countries maintained at NIFTS, Morioka, Japan, was screened with seven simple sequence repeat (SSR) markers to establish genetic identities, estimate genetic diversity, verify parentage and examine genetic relationships. All seven SSR loci produced one or two amplified fragments and made it possible to differentiate among the rootstocks, except between virus-free subclones and their original parents as well as between ‘Aotea 40’ and ‘Aotea 106’. The parentage of all but one rootstock clone was confirmed. The placement of individual rootstock clones into subgroups in a phenogram generated from un-weighted pair group average cluster analysis was generally based on parentage or genetic origin. Based upon the ability of the SSR data to differentiate these apple rootstocks and the high level of gene diversity, it is proposed that all commercial apple rootstocks be fingerprinted to provide a mechanism to verify the identity of individual clones.

Notes: : In the progressive freeze-concentration, limiting partition coefficient, K0, is a very important process parameter, which corresponds to the partition coefficient of a solute between the ice and liquid phase assuming the infinitesimal ice growth rate and/or the infinite mass-transfer rate at the ice-liquid interface. K0 was determined for glucose and various electrolytes in the single-component and multi-component systems. K0 was dependent both on the concentration and the type of solute. In the single-component system, osmotic pressure was proved to be the major determinant of K0. In the multi-component system, K0 for a solute was affected by the coexisting solute through the osmotic pressure of the total system. K0 was also dependent on viscosity when the change in the osmotic pressure was limited but the change in the viscosity was considerable because of the coexisting solute.

Notes: To determine whether hepatocellular apoptosis is involved in chronic liver injury in patients with chronic hepatitis C, the correlations among the rate of hepatocellular apoptosis, the rate of hepatocytes infected with hepatitis C virus (HCV) and TGF-β1 mRNA expression, were histologically examined. HCV was visualized by in situ PCR technique. Apoptotic hepatocytes were determined by TUNEL technique. TGF-β1 mRNA expression was quantified by Northern blotting analysis, and visualized by in situ hybridization. Fas-antigen and proliferating cell nuclear antigen (PCNA) on hepatocytes, and α-smooth muscle actin (αSMA) and desmin were evaluated by immunohistochemistry. In the areas of inflammation, TUNEL-positive apoptotic hepatocytes and PCNA-positive hepatocytes were occasionally observed around HCV-positive hepatocyte. TGF-β1 mRNA-positive myofibroblast-like Ito cells (double-positive for αSMA and desmin) were also observed near the HCV-positive hepatocytes. Fas-antigen was detected on the surface of most hepatocytes in all lobules. A high correlation was found between the rate of HCV positive hepatocytes and that of apoptotic hepatocytes (r = 0.81, P 〈 0.05). The increase in number of HCV-positive hepatocytes was linked to the increases in TGF-β1 mRNA level (r = 0.79, P 〈 0.05) and number of TGF-β1 mRNA-positive Ito cells (r = 0.74, P 〈 0.05). These findings confirmed that the increase in HCV-infected hepatocytes is related to the increase of hepatocellular apoptosis and the increase of TGF-β1 mRNA in the liver of hepatitis C patients. TGF-β1 plays an important role in apoptosis of hepatocytes infected with HCV.

Notes: Ultraviolet (UV) irradiation induces damage of the skin, and in particular, photoageing is known to be the result of chronic UV irradiation. Many investigations have attempted to clarify the mechanisms of photoageing induced by chronic UVA irradiation, but consensus has not been achieved yet by in vivo experiments, mostly due to differences among UV sources and animals used for experiments. In vitro experiments have shown that a single exposure to UVA irradiation causes overexpression of matrix metalloproteinases and denaturation of collagen, but the mechanisms of the photoageing effects of chronic UVA irradiation are still unclear. To examine the effects of chronic UVA irradiation, we used an in vitro fibroblast cellular ageing system as a model of photoageing. Chronic UVA irradiation of normal human fibroblasts induced shortening of the cellular life span and an increase of cellular diameter, in parallel with expression of senescence-associated β-galactosidase. Extracellular degradation enzyme, matrix metalloproteinase 1 (MMP-1) was overexpressed after repeated UVA irradiation, but tissue inhibitor of metalloproteinase 1 (TIMP-1) expression was hardly changed by chronic UVA irradiation. We conclude that chronic UVA irradiation of normal human fibroblasts induces cellular functional changes, leading to accelerated cellular ageing and MMP-1 overexpression.

Notes: MtDNA sequence data (c. 2100 bp of complete D-loop + cytochrome b gene regions) showed a principal phylogenetic split within common carp Cyprinus carpio between the Lake Biwa wild form (n = 3) and ‘Eurasian’ wild (n = 18) and domesticated (n = 35) forms. This supports the idea of the ancient origin of the Lake Biwa wild form, as well as the East Asian origin of wild common carp, previously proposed on the basis of fossil data. In addition, a possible multiple origin of koi carp was indicated by the polyphyletic distribution of five mtDNA haplotypes of koi carp within the ‘Eurasian’ clade.

Notes: Background:  Some studies suggest that apoptosis-related factors are involved in the inflammatory processes of marginal periodontal lesions. However, the role of apoptosis in periapical inflammatory lesions remains unclear. We investigated the possible role of apoptotic cell death in periapical inflammatory lesions by means of immunohistochemical analysis of apoptosis-related factors and use of a cell proliferation marker.Methods:  Paraffin-embedded sections of 19 radicular cysts (RCs), and five residual radicular cysts (RRCs) and control specimens of normal gingivae excised from seven cadavers were prepared and examined immunohistochemically with the use of monoclonal antibodies or polyclonal antisera against single-stranded DNA (ssDNA), p53, Bax, Bcl-2, caspase-3, Fas, Fas ligand (Fas-L), and Ki-67 antigen.Results:  Epithelium of gingiva, RCs, and RRCs showed expression of ssDNA in suprabasal and superficial epithelial cells and Ki-67 reactivity in basal and parabasal cells. Expression of Ki-67 and ssDNA in RCs and RRCs was slightly higher than that in gingiva. Both Ki-67 and ssDNA reactivity in RCs with intense inflammatory reactions or with thick lining epithelium were significantly stronger than those in RCs with less inflammatory reactions or with thin lining epithelium. Reactivity for p53 was noted sporadically in epithelium of gingiva, RCs, and RRCs, and p53 expression in RCs was significantly greater than that in gingiva. Ki-67 and ssDNA reactivity in RCs increased parallel to the degree of p53 expression. Bax and Bcl-2 were detected in some basal epithelial cells in RCs and RRCs as well as in gingiva. The ssDNA reactivity significantly increased parallel to Bax expression and slightly decreased parallel to Bcl-2 expression in lining epithelium of RCs. Caspase-3 was detected in superficial epithelial cells of both gingiva and lining epithelium of RCs and RRCs, and the distribution of these cells was compatible with the expression of ssDNA. Expression of Ki-67 and ssDNA in caspase-3-positive fields was significantly higher than that in caspase-3-negative fields in RCs. There was very limited expression of Fas and Fas-L in lining epithelium of RCs and RRCs as well as in gingiva.Conclusions:  These data suggest that apoptosis-related factors are involved in the pathophysiologic activity of periapical inflammatory lesions. Such factors may be affected by the structure of lining epithelium and the degree of inflammatory change.

Notes: A 49-year-old Japanese male first visited our clinic at May 13th, 2003. A bean-sized black macule had been noticed in the parietal region since childhood. The lesion had increased in size gradually and had become 30 mm in diameter. The lesion was diagnosed as malignant melanoma clinically and also dermoscopically. Sentinel node navigation using radioisotope revealed the metastases. We performed wide local excision and radical neck dissection, and the definite diagnosis of malignant melanoma and regional nodal metastases was made. TNM staging was stage?b, that is, pT4b, N2c, M0. The patient also had nine small black freckles on the trunk and extremities. All of them were excised, and eight lesions of them were diagnosed as malignant melanoma, because proliferation of atypical melanocytes and formation of abundant abnormal nests in epidermis were observed histopathologically. Chemotherapy was performed, but metastases of skin and visceral region were found in February, 2004. These lesions had put up strong resistance to chemotherapy and the patient died on June, 6th, 2004. Autopsy revealed multiple metastases including heart, kidneys, liver and large intestine and so on. No other family members suffered from malignant melanoma. The origin of this patient’s melanoma remained unclear.

Notes: Ultraprecision optical components require ultra-fine smooth surface quality of sub-nanometer or sub-angstrom in Ra. To satisfy these requirements, ultraprecision on-machine measurement system is very important, by which the profile measurement and evaluation is conducted on the machine. The form accuracy is improved by compensating machining when the form accuracy is not enough. Ultraprecision on-machine measurement systems were developed for measurement with high-accuracy and high-efficiency. It was confirmed that the on-machine measurement with AFM was possible for surface roughness or complex form for local areas. In addition, a laser probe unit with a maximum resolution of 1nm, a measuring range of 10mm, a repeatability of 5.6nm and a maximum measurable angle of 60 degrees was developed. The possibility of the non-contact on-machine form measurement was confirmed for global form accuracy control