Multi spectral holographic ellipsometry for a complex 3D nanostructure.

We present an innovative ellipsometry technique called self-interferometric pupil ellipsometry (SIPE), which integrates self-interference and pupil microscopy techniques to provide the high metrology sensitivity required for metrology applications of advanced semiconductor devices. Due to its unique configuration, rich angle-resolved ellipsometric information from a single-shot hologram can be extracted, where the full spectral information corresponding to incident angles from 0° to 70° with azimuthal angles from 0° to 360° is obtained, simultaneously. The performance and capability of the SIPE system were fully validated for various samples including thin-film layers, complicated 3D structures, and on-cell overlay samples on the actual semiconductor wafers. The results show that the proposed SIPE system can achieve metrology sensitivity up to 0.123 nm. In addition, it provides small spot metrology capability by minimizing the illumination spot diameter up to 1 µm, while the typical spot diameter of the industry standard ellipsometry is around 30 µm. As a result of collecting a huge amount of angular spectral data, undesirable multiple parameter correlation can be significantly reduced, making SIPE ideally suited for solving several critical metrology challenges we are currently facing.

Rhodobacter azotoformans LPS (RAP99-LPS) Is a TLR4 Agonist That Inhibits Lung Metastasis and Enhances TLR3-Mediated Chemokine Expression.

The lipopolysaccharides (LPSs) of Rhodobacter are reported to be TLR4 antagonists. Accordingly, the extract of Rhodobacter azotoformans (RAP99) is used as a health supplement for humans and animals in Japan to regulate immune responses in vivo. We previously analyzed the LPS structure of RAP99 (RAP99-LPS) and found it is different from that of E. coli-LPS but similar to lipid A from Rhodobacter sphaeroides (RSLA), a known antagonist of TLR4, with both having three C14 fatty acyl groups, two C10 fatty acyl groups, and two phosphates. Here we show that RAP99-LPS has an immune stimulatory activity and acts as a TLR4 agonist. Pretreatment of RAP99-LPS suppressed E. coli-LPS-mediated weight loss, suggesting it is an antagonist against E. coli-LPS like other LPS isolated from Rhodobacter. However, injections of RAP99-LPS caused splenomegaly and increased immune cell numbers in C57BL/6 mice but not in C3H/HeJ mice, suggesting that RAP99-LPS stimulates immune cells via TLR4. Consistently, RAP99-LPS suppressed the lung metastasis of B16F1 tumor cells and enhanced the expression of TLR3-mediated chemokines. These results suggest that RAP99-LPS is a TLR4 agonist that enhances the activation status of the immune system to promote anti-viral and anti-tumor activity in vivo.

Structural analysis of a novel lipooligosaccharide (LOS) from Rhodobacter azotoformans.

Lipopolysaccharides (LPS) are components of the Gram-negative bacterial cell surface that stimulate the host innate immune system through the Toll-like receptor (TLR) 4-MD-2 complex. Rhodobacter sp. have been reported to produce LPS that lack endotoxic activity, and instead act as antagonists of other endotoxins. In this report, we focused on LPS, especially the lipooligosaccharide (LOS) fraction produced by Rhodobacter azotoformans that shows production of IL-8, but has an inverse correlation with IL-6 production. We analyzed their molecular structure by using mass spectrometry and nuclear magnetic resonance spectroscopy and report a novel LOS consisting of a shorter glycan structure containing glucuronic acid but not heptoses. A novel glycan structure, Glcα(1 → 4)GlcAα(1 → 4)KDOα(2 → 4)[Glcα(1 → 5)]KDOα(2 → 6)[4-phosphate]GlcNß(1 → 6) GlcNα1-phosphate, was proposed using NMR methods. The structure was consistent with one obtained based on MS. The MS analysis further revealed the existence of structural variation caused by extension with hexoses. The acyl composition in lipid A was suggested to contain three C14 fatty acyl chains (3-OH-14:0 or 3-oxo-14:0 at N2 of GlcN-1, 3-OH-14:0 at N2 of GlcN-2, that carried another 14:1 Δ7 on its ß-hydroxyl group) and two C10 fatty acyl chains (3-OH-10:0 at O3 of both GlcN), which are same as those found in lipid A from Rhodobacter sphaeroides.

Mutation spectrum in the Wnt/ß-catenin signaling pathway in gastric fundic gland-associated neoplasms/polyps.

Frequent activation of the Wnt/ß-catenin signaling pathway has recently been demonstrated in gastric adenocarcinoma/neoplasia of chief cell predominant type (GA-CCP/GN-CCP) with submucosal involvement. In this study, we examined the activation status of the Wnt/ß-catenin signaling pathway in GN-CCP without submucosal involvement, which is referred to as gastric dysplasia-CCP (GD-CCP). We also examined ß-catenin expression and the mutation spectrum of PPP2R1A and Wnt pathway genes in 11 cases of GD-CCP, 25 cases of gastric polyps of fundic gland type (GPs-FG), and 21 cases of GPs-FG with dysplasia (GP-FGD). ß-catenin nuclear staining was observed in 3 cases of GD-CCP, none of GPs-FG, and 6 cases of GPs-FGD. Mutations in Wnt pathway genes, including PPP2R1A, were observed in 4 cases of GDs-CCP, 10 cases of GPs-FG, and 7 cases of GPs-FGD. Two of these seven GPs-FGD cases showed ß-catenin nuclear staining. However, none of the 4 GD-CCP cases with mutations or the 10 GPs-FG cases with mutations showed ß-catenin nuclear staining. PPP2R1A mutations were observed in 1 GD-CCP case and 1 GPs-FGD case. Although the mutation spectra of the Wnt pathway genes in GD-CCP and GP-FG differed, based on the absence of ß-catenin nuclear staining despite the genetic alterations, GD-CCP is more similar to GP-FG than to GN-CCP, which shows ß-catenin nuclear staining and submucosal involvement. Activation of the Wnt/ß-catenin signaling by the ß-catenin nuclear transition may be required during progression from GD-CCP to GN-CCP. Furthermore, this is the first report describing PPP2R1A mutations in gastric fundic gland-associated neoplasms.

GNAS mutation as an alternative mechanism of activation of the Wnt/ß-catenin signaling pathway in gastric adenocarcinoma of the fundic gland type.

Gastric adenocarcinoma of the fundic gland type (GAFG) is a rare variant of gastric tumor. We have recently reported the frequent accumulation of ß-catenin in GAFGs and showed that approximately half of the cases studied harbored at least 1 mutation in CTNNB1/AXINs/APC, leading to the constitutive activation of the Wnt/ß-catenin pathway. However, the mechanisms of Wnt signaling activation in the remaining cases are unknown. Accumulating evidence showed that the activating mutation in GNAS promotes tumorigenesis via the activation of the Wnt/ß-catenin pathway or the ERK1/2 MAPK pathway. Therefore, we analyzed the mutations in GNAS (exons 8 and 9) and in KRAS (exon 2) in 26 GAFGs. Immunohistochemistry revealed nuclear ß-catenin expression in 22 of 26 GAFGs, and 10 (38.5%) of 26 cases harbored at least 1 mutation in CTNNB1/AXINs/APC. Activating mutations in GNAS were found in 5 (19.2%) of 26 GAFGs, all of which harbored R201C mutations. Activating mutations in KRAS were found in 2 (7.7%) of 26 GAFGs, and both of these also contained GNAS activating mutations. Four of 5 cases with GNAS mutation showed nuclear ß-catenin expression, and presence of GNAS mutation was associated with ß-catenin nuclear expression (P = .01). Furthermore, 3 of these 4 cases did not harbor mutations in CTNNB1, APC, or AXINs, suggesting that mutations in the Wnt component genes and those in GNAS occur almost exclusively. These results suggest that GNAS mutation might occur in a small subset of GAFG as an alternative mechanism of activating the Wnt/ß-catenin signaling pathway.

Two cases of proximal pole scaphoid fracture accompanied by lunate fracture.

We experienced two cases of simultaneous fracture of the scaphoid and the lunate. In two cases, both scaphoid and lunate fractures existed on the same plane, which may help us to understand the mechanism of proximal fracture of the scaphoid.

Alteration in the Wnt/ß-catenin signaling pathway in gastric neoplasias of fundic gland (chief cell predominant) type.

Gastric neoplasia of chief cell-predominant type (GN-CCP) has been reported as a new, rare variant of gastric tumor. GN-CCPs were defined as tumors consisting of irregular anastomosing glands of columnar cells mimicking chief cells of fundic gland with nuclear atypia and prolapse-type submucosal involvement. We comparatively evaluated clinicopathologic features between 31 GN-CCPs and 130 cases of conventional gastric adenocarcinoma invading into submucosa (CGA-SM) in addition to nuclear ß-catenin immunolabeling and direct sequencing of members of the Wnt/ß-catenin pathway, CTNNB1, APC, and AXIN, in a subset of these tumors. GN-CCP presented as small protruded lesions located in the upper third of the stomach, with minimal involvement into the submucosa and rare lymphovascular invasion. None of the lesions have demonstrated a recurrence of disease or metastasis on follow-up. Nuclear ß-catenin immunolabeling was higher in GN-CCP (labeling index [LI]: median, 19.3%; high expresser [LI >30%], 7/27 cases [26%]) than CGA-SM (median LI, 14.7%; high expresser, 1/19 cases [6%]). Missense mutation of APC was observed in 1 GN-CCP but not CGA-SM. Missense or nonsense mutations of CTNNB1 and AXIN1 were higher in GN-CCPs (14.8%, both) than CGA-SMs (5.3%, both). Missense mutations of AXIN2 were higher in GN-CCPs (25.9%) than in CGA-SMs (10.5%). Overall, 14 (51.9%) of 27 GN-CCPs and 5 (26.3%) of 19 CGA-SM cases harbored at least 1 of these gene mutations. In conclusion, GN-CCPs as a unique variant of nonaggressive tumor are characterized by nuclear ß-catenin accumulation and mutation of CTNNB1 or AXIN gene, suggesting activation of the Wnt/ß-catenin pathway.

MUC5AC/ß-catenin expression and KRAS gene alteration in laterally spreading colorectal tumors.

AIM: To clarify differences in mucin phenotype, proliferative activity and oncogenetic alteration among subtypes of colorectal laterally spreading tumor (LST). METHODS: LSTs, defined as superficial elevated lesions greater than 10 mm in diameter with a low vertical axis, were macroscopically classified into two subtypes: (1) a granular type (Gr-LST) composed of superficially spreading aggregates of nodules forming a flat-based lesion with a granulonodular and uneven surface; and (2) a non-granular type (NGr-LST) with a flat smooth surface and an absence of granulonodular formation. A total of 69 LSTs, comprising 36 Gr-LSTs and 33 NGr-LSTs, were immunohistochemically stained with MUC2, MUC5AC, MUC6, CD10 (markers of gastrointestinal cell lineage), p53, ß-catenin and Ki-67 antibodies, and examined for alteration in exon 1 of v-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog (KRAS) and exon 15 of v-raf murine sarcoma viral oncogene homologue B1 (BRAF) by polymerase chain reaction followed by direct sequencing. RESULTS: Histologically, 15 Gr-LST samples were adenomas with low-grade dysplasia (LGD), 12 were high-grade dysplasia (HGD) and 9 were adenocarcinomas invading the submucosa (INV), while 12 NGr-LSTs demonstrated LGD, 14 HGD and 7 INV. In the proximal colon, MUC5AC expression was significantly higher in the Gr-type than the NGr-type. MUC6 was expressed only in NGr-LST. MUC2 or CD10 did not differ. P53 expression demonstrated a significant stepwise increment in progression through LGD-HGD-INV with both types of LST. Nuclear ß-catenin expression was significantly higher in the NGr-type. Ki-67 expression was significantly higher in the Gr-type in the lower one third zone of the tumor. In proximal, but not distal colon tumors, the incidence of KRAS provided mutation was significantly higher in the Gr-type harboring a specific mutational pattern (G12V). BRAF mutations (V600E) were detected only in two Gr-LSTs. CONCLUSION: The two subtypes of LST, especially in the proximal colon, have differing phenotypes of gastrointestinal cell lineage, proliferation and activation of Wnt/ß-catenin or RAS/RAF/extracellular signal-regulated kinase signaling.

Error analysis and compensation method of focus detection in exposure apparatus.

An original approach to measurement accuracy of a typical focus sensor in conventional integrated circuit lithographic equipment is introduced. Causes of measurement error in the focus sensor are theoretically analyzed and found to be generated mainly from interactions between imperfections of the optical system and the actual surface of processed wafers. We derive mathematical formulations describing these errors, which are confirmed by the experimental results performed by using an optical setup composed of the focus sensor and samples on which the wafer surface condition is reproduced. Furthermore, several novel techniques that are intended to reduce those measurement errors are successfully demonstrated.

Three-dimensional magnetic resonance imaging after ultrasonography for assessment of fetal gastroschisis.

A 24-year-old woman (Gravida I, Para I) at estimated 32 weeks of pregnancy was referred to our department for evaluation of a suspected fetal gastroschisis. Ultrasound scan revealed multiple loops of dilated bowel outside the fetal abdomen and absence of membrane surrounding the herniated loops of the intestines. Three-dimensional (3D) magnetic resonance imaging was performed to obtain more information on the bowel both outside and inside the abdomen. Images were constructed with T1-weighted fat-suppressed 3D fast low-angle shot sequences using a maximum intensity projection algorithm. The 3D images made possible the realization of fetal bowel conditions with greater definition and accuracy.

X-ray telescope onboard Astro-E. III. Guidelines to performance improvements and optimization of the ray-tracing simulator.

We present a detailed study of the performance of the Astro-E x-ray telescope (XRT) onboard the Astro-E satellite. As described in preceding papers the ground-based calibrations of the Astro-E XRT revealed that its image quality and effective area are somewhat worse than that expected from the original design. Conceivable causes for such performance degradation are examined by x-ray and optical microscopic measurements at various levels, such as individual reflectors, sectors, and quadrants of the XRT and their alignments. We can attribute, based on detailed measurements, the degradation of the image quality to a slope error in the individual reflectors and the positioning error of reflectors. As for the deficit of the effective area, the shadowing of x rays within the XRT body is the dominant factor. Error budgets for the performance degradation of the Astro-E XRT are summarized. The ray-tracing simulator, which is needed to construct the response function for arbitrary off-axis angles and spatial distributions of any celestial x-ray sources, has been developed and tuned based on the results of detailed measurements. The ray-tracing simulation provides results that are consistent within 3% with the real measurement except for large off-axis angles and higher energies. We propose, based on knowledge obtained from all the measurements and simulations, several plans for future developments to improve the performance of the nested thin-foil mirrors.