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Patient-specific guides have been introduced recently as a means of making accurate bone cuts through custom cutting blocks constructed based on pre-operative three-dimensional imaging. However, the controversy concerning the improved results of patient-specific guides have not been resolved yet; in addition, there have been no studies to investigate the causes of variable with inconsistent results and solutions for the causes.Thirty eight patients (38 knees) underwent total knee arthroplasty with patient-specific guides. The mean age of the patients was 68 years (SD ± 6.3), and all patients had a minimum 2-year follow-up. An intra-operative alignment using navigation and the causes of outliers were evaluated.An average coronal alignment of PSI jigs was 0.5° (SD ± 0.9°) in femur and 0.1° (SD ± 0.8°) in tibia, and the number of outliers was two and three cases, respectively. An average sagittal alignment was 0.6° (SD ± 0.9°) in femur and 5.5° (SD ± 1.1°) in tibia, and the number of outliers was three and five cases, respectively. All outliers resulted from large osteophytes near the contact point of patient-specific guides which disturb sitting of the guide.It was suggested that patient-specific guides were an effective and safe method to achieve accurate alignments, with no additional intra-operative complication. It is important to note that surgeons need to be precautious using the patient-specific instrumentation in patients with severe varus deformity. In addition, existing osteophytes which disturb sitting of the guides should be carefully evaluated pre-operatively and intra-operatively.IV.

Although the role of secretory granules as the inositol 1,4,5-trisphosphate (IP(3))-sensitive intracellular Ca(2+) store and the presence of the IP(3) receptor (IP(3)R)/Ca(2+) channel on the secretory granule membrane have been established, the identity of the IP(3)R types present in the secretory granules is not known. We have therefore investigated the presence of different types of IP(3)R in the secretory granules of bovine adrenal medullary chromaffin cells using immunogold electron microscopy and found the existence of all three types of IP(3)R in the secretory granules. To determine whether these IP(3)Rs interact with CGA and CGB, each IP(3)R isoform was co-transfected with CGA or CGB into NIH3T3 or COS-7 cells, and the expressed IP(3)R isoform and CGA or CGB were co-immunoprecipitated. From these studies it was shown that all three types of IP(3)R form complexes with CGA and CGB in the cells. To further confirm whether the IP(3)R isoforms and CGA and CGB form a complex in the secretory granules the potential interaction between all three isoforms of IP(3)R and CGA and CGB was tested by co-immunoprecipitation experiments of the mixture of secretory granule lysates and the granule membrane proteins. The three isoforms of IP(3)R were shown to form complexes with CGA and CGB, indicating the complex formation between the three isoforms of IP(3)R and CGA and CGB in the secretory granules. Moreover, the pH-dependent Ca(2+) binding property of CGB was also studied using purified recombinant CGB, and it was shown that CGB bound 93 mol of Ca(2+)/mol with a dissociation constant (K(d)) of 1.5 mm at pH 5.5 but virtually no Ca(2+) at pH 7.5. The high capacity, low affinity Ca(2+)-binding property of CGB at pH 5.5 is comparable with that of CGA and is in line with its role as a Ca(2+) storage protein in the secretory granules.

The secretory granules of neuroendocrine cells which contain large amounts of Ca(2+) and chromogranins have been demonstrated to release Ca(2+) in response to inositol 1,4,5-trisphosphate (IP(3)). Moreover, chromogranin A (CGA) has been shown to interact with several secretory granule membrane proteins, including the IP(3) receptor (IP(3)R). To determine whether the IP(3)Rs interact directly with chromogranins A and B (CGB), two major proteins of the secretory granules, we have used purified IP(3)R from bovine cerebellum in the interaction study with CGA and CGB, and have shown that chromogranins A and B directly interact with the IP(3)R at the intravesicular pH 5.5. Immunogold cytochemical study using the IP(3)R and CGA antibodies indicated that IP(3)R-labeled gold particles were localized in the periphery of the secretory granules, indicating the presence of the IP(3)Rs on the secretory granule membrane. To determine whether the IP(3)R and chromogranins A and B are physically linked in the cells, bovine type 1 IP(3)R (IP(3)R-1) and CGA or CGB are co-transfected into COS-7 cells and co-immunoprecipitation was carried out. Immunoprecipitation of the cell extracts demonstrated the presence of CGA-IP(3)R-1 and CGB-IP(3)R-1 complexes, respectively, indicating the complex formation between the IP(3)R and chromogranins A and B in native state.

Temperature dependence of elastic constants of thin cold-rolled stainless steel has been measured by using the acoustic resonance method. Identification of the vibration mode has been examined numerically and experimentally. The elastic constants at room temperature have also been measured by the pulse echo method. In addition, the texture effect on the elastic constants has been analysed by assuming the specimen has orthorhombic structure.

Catecholamine secretory vesicle core proteins (chromogranins) contain an activity that inhibits catecholamine release, but the identity of the responsible peptide has been elusive. Size-fractionated chromogranins antagonized nicotinic cholinergic-stimulated catecholamine secretion; the inhibitor was enriched in processed chromogranin fragments, and was liberated from purified chromogranin A. Of 15 synthetic peptides spanning approximately 80% of chromogranin A, one (bovine chromogranin A344-364 [RSMRLSFRARGYGFRGPGLQL], or catestatin) was a potent, dose-dependent (IC50 approximately 200 nM), reversible secretory inhibitor on pheochromocytoma and adrenal chromaffin cells, as well as noradrenergic neurites. An antibody directed against this peptide blocked the inhibitory effect of chromogranin A proteolytic fragments on nicotinic-stimulated catecholamine secretion. This region of chromogranin A is extensively processed within chromaffin vesicles in vivo. The inhibitory effect was specific for nicotinic cholinergic stimulation of catecholamine release, and was shared by this chromogranin A region from several species. Nicotinic cationic (Na+, Ca2+) signal transduction was specifically disrupted by catestatin. Even high-dose nicotine failed to overcome the inhibition, suggesting noncompetitive nicotinic antagonism. This small domain within chromogranin A may contribute to a novel, autocrine, homeostatic (negative-feedback) mechanism controlling catecholamine release from chromaffin cells and neurons.

Since the conserved near N-terminal region (residues 18-37) of chromogranin A (CGA) has tentatively been identified as the vesicle membrane interacting region using synthetic peptides, it was necessary to confirm this finding with CGA deletion proteins. In order to address this need and to clarify the discrepancies of the published amino acid sequences of CGA, we cloned a CGA gene and produced CGA deletion proteins of various sizes. The recombinant CGA protein lacking the first 16 amino acid residues bound to the vesicle membrane as well as the full-length CGA at pH 5.5. However, the CGA protein lacking the first 39 amino acid residues, which include the conserved near N-terminal region (residues 17-38), failed to interact with the vesicle membrane at pH 5.5, clearly indicating the essential role of the conserved near N-terminal region in the pH-dependent interaction of CGA with the vesicle membrane.

Mitochondrial dysfunction in dopaminergic neurons of patients with idiopathic and familial Parkinson's disease (PD) is well known although the underlying mechanism is not clear. We established a homogeneous population of human adipose tissue-derived mesenchymal stromal cells (hAD-MSCs) from human adult patients with early-onset hereditary familial Parkin-defect PD as well as late-onset idiopathic PD by immortalizing cells with the hTERT gene to better understand the underlying mechanism of PD. The hAD-MSCs from patients with idiopathic PD were designated as "PD", from patients with Parkin-defect PD as "Parkin" and from patients with pituitary adenomas as "non-PD" in short. The pGRN145 plasmid containing hTERT was introduced to establish telomerase immortalized cells. The established hTERT-immortalized cell lines showed chromosomal aneuploidy sustained stably over two-years. The morphological study of mitochondria in the primary and immortalized hAD-MSCs showed that the mitochondria of the non-PD were normal; however, those of the PD and Parkin were gradually damaged. A striking decrease in mitochondrial complex I, II, and IV activities was observed in the hTERT-immortalized cells from the patients with idiopathic and Parkin-defect PD. Comparative Western blot analyses were performed to investigate the expressions of PD specific marker proteins in the hTERT-immortalized cell lines. This study suggests that the hTERT-immortalized hAD-MSC cell lines established from patients with idiopathic and familial Parkin-defect PD could be good cellular models to evaluate mitochondrial dysfunction to better understand the pathogenesis of PD and to develop early diagnostic markers and effective therapy targets for the treatment of PD.

Chromogranins A and B (CGA and CGB), the major proteins of the secretory vesicles of the regulated secretory pathway, have been shown to aggregate in a low pH and high calcium environment, the condition found in the trans-Golgi network where secretory vesicles are formed. Moreover, CGA and CGB, as well as several other secretory vesicle matrix proteins, have recently been shown to bind to the vesicle membrane at the intravesicular pH of 5.5 and to be released from it at a near physiological pH of 7.5. The pH- and Ca-dependent aggregation and interaction of chromogranins, as well as several other matrix proteins, with the vesicle membrane are considered essential in vesicle biogenesis. Therefore, to gain further insight into how vesicle matrix proteins find their way into the secretory vesicles, the pH- and Ca-dependent aggregation and vesicle membrane binding properties of the vesicle matrix proteins were studied, and it was found that most of the vesicle matrix proteins aggregated in the presence of Ca at the intravesicular pH of 5.5. Furthermore, most of the vesicle matrix proteins bound not only to the vesicle membrane but also to CGA at pH 5.5, with the exception of a few matrix proteins that appeared to bind only to CGA or to vesicle membrane. Purified CGB was also shown to interact with CGA at pH 5.5. The extent and Ca-sensitivity of the aggregation of vesicle matrix proteins lay between those of purified CGB and CGA, CGB aggregation showing the highest degree of aggregation and being the most Ca sensitive at a given protein concentration. Hence, in view of the abundance of chromogranins in secretory vesicles and their low pH- and high calcium-dependent aggregation property, combined with their ability to interact with both the vesicle matrix proteins and the vesicle membrane, CGA and CGB are proposed to play essential roles in the selective aggregation and sorting of potential vesicle matrix proteins to the immature secretory vesicles of the regulated secretory pathway.

Chromogranins A and B have been known to undergo pH- and Ca(2+)-dependent aggregation, and this property is considered essential for the proper sorting of the vesicular matrix proteins. In the present study, purified native chromogranin B (CGB) from bovine adrenal medulla was used to study the pH- and Ca(2+)-dependent conformational changes and aggregation property. Similar to chromogranin A (CGA), which had been shown to undergo pH- and Ca(2+)-dependent conformational changes and to be composed of 60-65% random coil with 25-40% alpha-helicity, chromogranin B was also shown to consist of 65-70% random coil, 15-25% alpha-helix, and 10-15% beta-sheet structures. The high percentage of random coil suggests that CGB behaves hydrodynamically as an asymmetric molecule, thus explaining its anomalous migration on SDS-polyacrylamide gels. Further, CGB eluted from a gel filtration column in the volume indicative of a globular protein with molecular weight of approximately 200,000 at both the intravesicular pH of 5.5 and a near physiological pH of 7.5. Considering that dimeric CGA eluted from a gel filtration column in the position suggestive of a 300-kDa protein, this result indicated that CGB exists in a monomeric state at both pH levels. Like CGA, which exhibited greater aggregation at pH 5.5 than at pH 7.5 upon Ca2+ binding, CGB also aggregated much more readily at pH 5.5 than at pH 7.5. However, there was a marked difference in the aggregation properties of CGA and CGB with regard to their sensitivity to Ca2+: CGB was at least 2 orders of magnitude more sensitive to Ca2+ than CGA. This suggested that, in spite of the low concentration of CGB (approximately one-tenth that of CGA) in bovine adrenal chromaffin cells, CGB would start to aggregate well ahead of CGA in the trans-Golgi network. In view of the proposed importance of the pH- and Ca(2+)-induced chromogranin aggregation in vesicle biogenesis, the extreme sensitivity of CGB aggregation to Ca2+ appears to underline the potential importance of CGB aggregation in the early stages of vesicle biogenesis.

Seung-Hyun Yoo, Hyuk-Ju Kwon, Sun-Hee Kim, Jung-Ah Lee, Sung Sunwoo, Young-Sik Kim, Byung-Su Kim, Han-Jin Oh, Chul-Min Kim, Hee-Jeong Choi, Hee-Jin Hwang, Sun-Wha Ok Department of Family Medicine, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea Department of Psychiatry, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea Department of Family Medicine, Vievis Namuh Hospital, Seoul, Korea Department of Family Medicine, Seoul St. Mary's Hospital, The Catholic University of Korea School of Medicine, Seoul, Korea Department of Family Medicine, Eulji University School of Medicine, Daejeon, Korea Department of Family Medicine, Catholic Kwandong University International St. Mary's Hospital, Catholic Kwandong University College of Medicine, Seoul, Korea Department of Child Development and Family Studies, Institute of Human Ecology, Seoul National University, Seoul, Korea

암의 조기발견과 암 치료의 발전에 따라 암 경험자의 수가 빠르게 증가하고 있다. 2015년 발표된 중앙암등록본부 암 통계에 따르면 암 진단 후 현재까지 생존해 있는 환자는 137 만 명이 넘어서고 있으며, 5년을 초과하여 생존한 암 경험자 의 수도 58만 5,000명에 달하고 있다[1]. 장기 생존한 암 경 험자들이 증가하면서, 암 치료를 시행하는 암 전문의료기관 뿐 아니라 지역사회의 일차의료기관에서도 암 경험자들을 보는 것이 점점 흔해지고 있다. 특히 위암은 우리나라에서 전통적으로 흔했던 암으로, 암 치료 후 5년 이상 경과한 암 경험자의 수는 2013년 말 현재 11만 명으로 모든 암 중 가장 많은 수를 차지하고 있다[1]. 암 경험자는 원발암에 대한 추적관리뿐 아니라, 이차암에 대한 예방 및 검진, 후기 합병증 및 암 이외의 동반 만성질 환에 대한 관리, 예방접종, 생활습관, 심리사회적 문제에 대 한 포괄적인 관리가 필요하다. 본고에서는 일차의료기관에 서 보게 될 수 있는 위암 치료 후 장기간 경과한 위암 경험 자의 흔한 건강문제와 이에 대한 관리방안을 다루고자 한다. 장기 위암 경험자의 관리

We propose a vacuum wafer-level packaging (WLP) process using glass-reflowed silicon via for nano/micro devices (NMDs). A through-wafer interconnection (TWIn) substrate with silicon vias and reflowed glass is introduced to accomplish a vertical feed-through of device. NMDs are fabricated in the single crystal silicon (SCS) layer which is formed on the TWIn substrate by Au eutectic bonding including Cr adhesion layer. The WLPof the devices is achieved with the capping glass wafer anodically bonded to the SCS layer. In order to demonstrate the successful hermetic packaging, we fabricated the micro-Pirani gauge in the SCS layer, and packaged it in the wafer-level. The vacuum level inside the packaging was measured to be 3.1 Torr with +/- 0.12 Torr uncertainty, and the packaging leakage was not detected during 24 hour after the packaging.

Chromogranin A is a high capacity, low affinity Ca(2+)-binding protein suggested to be responsible for the Ca2+ storage function of the secretory vesicle, which has been identified as a major inositol 1,4,5-trisphosphate (IP3)-sensitive intracellular Ca2+ store of adrenal medullary chromaffin cells. Moreover, chromogranin A has recently been shown to interact with the vesicle membrane at the intravesicular pH of 5.5 and to be released from it at a near physiological pH of 7.5 (Yoo, S. H. (1993) Biochemistry 32, 8213-8219). In the present study, chromogranin A is shown to interact with several integral membrane proteins of secretory vesicles at pH 5.5 but not at pH 7.5. One of the chromogranin A-interacting membrane proteins had a mass of 260 kDa and reacted with the IP3 receptor antibody. This result suggested not only the existence of the IP3 receptor in the vesicle membrane but also the existence of direct communication between chromogranin A and the IP3 receptor. In addition, the pH-dependent interaction of chromogranin A with integral membrane proteins implies an important role for chromogranin A in the sorting process of the vesicle membrane proteins during vesicle biogenesis in the trans-Golgi network.

Chromogranin A is known to undergo pH induced conformational changes, and the difference in conformation is supposed to be responsible for the difference in Ca2+ binding property. To gain insight regarding the overall structure and the nature of pH-induced conformational changes of chromogranin A, limited trypsin digestions were carried out at pH 5.5 and pH 7.5. The resulting fragments were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and the amino acid sequences of the tryptic fragments were determined. From these analyses it was shown that the chromogranin A structure consists of an N-terminal compact core region and a rather loosely organized C-terminal region and that the change of pH from 7.5 to 5.5 loosened the overall structure of chromogranin A, exposing the C-terminal region. Since the conserved C-terminal region (residues 407–431) was shown to exist in monomer-dimer and monomer-tetramer equilibria at pH 7.5 and 5.5, respectively, the conformational changes of the region at pH 7.5 and 5.5 were studied by circular dichroism spectroscopy using a synthetic peptide representing the conserved C-terminal region. When the pH was changed from 7.5 to 5.5, the coil structure of the C-terminal peptide decreased with an accompanying increase of α-helicity.

Contrary to the notion that the soluble intravesicular matrix proteins of the secretory vesicles of adrenal medullary chromaffin cells freely float in the vesicle, several vesicle matrix proteins of the secretory vesicles, including chromogranins A and B, bound to the vesicle membrane at intravesicular pH (5.5) and were freed from it when the pH was raised to a near physiological pH (7.5). Estimation of the fraction of vesicle matrix proteins that might remain bound to the vesicle membrane in the vesicle suggested that the majority (> 50-80%) of chromogranins A and B, as well as several other proteins, will stay bound to the membrane in the vesicle. Comparison of the amino-acid sequences of chromogranins A and B revealed two highly conserved regions, i.e., one near the N-terminus and the other being the C-terminal region. Since it has been demonstrated with chromogranin A that the conserved near N-terminal region of chromogranin A exhibited the pH-dependent membrane-binding activity (Yoo, S. H. (1993) Biophys. J., 64, A195), the same region in chromogranin B (residues 17-36) was tested using a synthetic chromogranin B peptide, and found to exhibit the pH-dependent membrane-binding activity. The pH-dependent binding of the matrix proteins at pH 5.5 and the automatic untethering at a physiological pH accord well with the rapid release and circulation of the vesicular contents in the bloodstream.

Background: Diabetes self-management education and reinforcement are important for effective management of the disease. We investigated the effectiveness of interactive small-group education on glycemic, blood pressure, and lipid levels. Methods: For this study, 207 type 2 diabetes patients with suboptimal glycemic control (HbA1c levels >6.5%) were enrolled. The conventional education group received an existing education program from April to November in 2006, and the interactive education group received a new small-group education program from December 2006 to July 2007. The two groups were com-paratively analyzed for changes in blood sugar, glycated hemoglobin, lipid, and blood pressure at baseline, 3, 6, and 12 months and the proportion of patients achieving target goals at 12 months. Results: After 12 months of follow-up, HbA1c levels in the interactive education group were significantly lower than in the con-ventional education group (6.7% vs. 6.4%, P

During development, sympathetic neurons and chromaffin cells originate from bipotential sympathoadrenal (SA) progenitors arising from neural crests (NC) in the trunk regions. Recently, we showed that AP-2β, a member of the AP2 family, plays a critical role in the development of sympathetic neurons and locus coeruleus and their norepinephrine (NE) neurotransmitter phenotype. In the present study, we investigated the potential role of AP-2β in the development of NC-derived neuroendocrine chromaffin cells of the adrenal medulla and the epinephrine (EPI) phenotype determination. In support of its role in chromaffin cell development, AP-2β is prominently expressed in both embryonic and adult adrenal medulla. In adrenal chromaffin cells of the AP-2β(-/-) mouse, the expression levels of catecholamine biosynthesizing enzymes, dopamine β-hydroxylase (DBH) and phenylethanolamine-N-methyl-transferase (PNMT), as well as the SA-specific transcription factor, Phox2b, are significantly reduced compared to wild type. In addition, ultrastructural analysis demonstrated that the formation of large secretory vesicles, a hallmark of differentiated chromaffin cells, is defective in AP-2β(-/-) mice. Furthermore, the level of EPI content is largely diminished (>80%) in the adrenal gland of AP-2β(-/-) mice. Chromatin immunoprecipitation (ChIP) assays of rat adrenal gland showed that AP-2β binds to the upstream promoter of the PNMT gene in vivo; strongly suggesting that it is a direct target gene. Overall, our data suggest that AP-2β plays critical roles in the epinephrine phenotype and maturation of adrenal chromaffin cells.

The inositol 1,4,5-trisphosphate (IP(3))-mediated intracellular Ca(2+) releases in secretory cells play vital roles in controlling not only the intracellular Ca(2+) concentrations but also the Ca(2+)-dependent exocytotic processes. Of intracellular organelles that release Ca(2+) in response to IP(3), secretory granules stand out as the most prominent organelle and are responsible for the majority of IP(3)-dependent Ca(2+) releases in the cytoplasm of chromaffin cells. Bovine chromaffin granules were the first granules that demonstrated the IP(3)-mediated Ca(2+) release as well as the presence of the IP(3) receptor (IP(3)R) in granule membranes. Secretory granules contain all three (type 1, 2, and 3) IP(3)R isoforms, and 58-69% of total cellular IP(3)R isoforms are expressed in bovine chromaffin granules. Moreover, secretory granules contain large amounts (2-4 mM) of chromogranins and secretogranins; chromogranins A and B, and secretogranin II being the major species. Chromogranins A and B, and secretogranin II are high-capacity, low-affinity Ca(2+) binding proteins, binding 30-93 mol of Ca(2+)/mol of protein with dissociation constants of 1.5-4.0 mM. Due to this high Ca(2+) storage properties of chromogranins secretory granules contain ~40 mM Ca(2+). Furthermore, chromogranins A and B directly interact with the IP(3)Rs and modulate the IP(3)R/Ca(2+) channels, i.e., increasing the open probability and the mean open time of the channels 8- to 16-fold and 9- to 42-fold, respectively. Coupled chromogranins change the IP(3)R/Ca(2+) channels to a more ordered, release-ready state, whereby making the IP(3)R/Ca(2+) channels significantly more sensitive to IP(3).

Ca2+ signaling is an important determining factor in many cellular processes, especially in cancer cell proliferation, motility and invasion. Glioblastoma is the deadliest brain cancer with its average survival time of less than a year, with the most prominent cellular feature being the ability of these cells to migrate to and invade the neighboring tissue. We hypothesized that disturbing the Ca2+ signaling pathway would decrease the propensity for these cells to migrate. Thus, we investigated the detailed Ca2+ signaling pathway of the glioblastoma cells in response to various receptor tyrosine kinases (RTK) and G-protein coupled receptor (GPCR) agonists. Here we report that caffeine, which is a well-known activator of ryanodine receptors (RyRs), paradoxically inhibits inositol-1, 4, 5-triphospate receptor(IP3R)-mediated Ca2+ increase by selectively targeting IP3R subtype 3(IP3R3), whose mRNA expression is significantly increased in glioblastoma cells. Consequently, by inhibiting IP3R3-mediated Ca2+ release, caffeine was found to inhibit the invasion and migration of various glioblastoma cell lines in scrape motility, Matrigel invasion, soft agar, and brain slice implantation assays. In a mouse xenograft model of glioblastoma, caffeine intake via drinking water greatly increased mean survival duration of subject animals. These findings propose IP3R3 as a novel target for glioblastoma treatment and that caffeine may be a useful adjunct therapy that slows glioblastoma invasion and migration by selectively targeting IP3R3.

Background The gliotransmitters released from astrocytes are deemed to play key roles in the glial cell-neuron communication for normal function of the brain. The gliotransmitters, such as glutamate, ATP, D-serine, neuropeptide Y, are stored in vesicles of astrocytes and secreted following the inositol 1,4,5-trisphosphate (IP3)-induced intracellular Ca2+ releases. Yet studies on the identity of the IP3-dependent intracellular Ca2+ stores remain virtually unexplored. Principal Findings We have therefore studied the potential existence of the IP3-sensitive intracellular Ca2+ stores in the cytoplasm of astrocytes using human brain tissue samples in contrast to cultured astrocytes that had primarily been used in the past. It was thus found that secretory granule marker proteins chromogranins and secretogranin II localize in the large dense core vesicles of astrocytes, thereby confirming the large dense core vesicles as bona fide secretory granules. Moreover, consistent with the major IP3-dependent intracellular Ca2+ store role of secretory granules in secretory cells, secretory granules of astrocytes also contained all three (types 1, 2, and 3) IP3R isoforms. Significance Given that the secretory granule marker proteins chromogranins and secretogranin II are high-capacity, low-affinity Ca2+ storage proteins and chromogranins interact with the IP3Rs to activate the IP3R/Ca2+ channels, i.e., increase both the mean open time and the open probability of the channels, these results imply that secretory granules of astrocytes function as the IP3-sensitive intracellular Ca2+ store.

Chromogranin A is a high capacity, low affinity Ca2+ binding protein which undergoes Ca2+- and pH-dependent conformational changes, and has recently been suggested to play a Ca2+-buffering role in the secretory vesicle of adrenal medullary chromaffin cell, the major inositol 1,4,5-trisphosphate-sensitive intracellular Ca2+ store of chromaffin cell (Yoo, S.H., and Albanesi, J.P. (1990) J. Biol. Chem. 265, 13446-13448). In the present study, it is shown that chromogranin A exists in a monomer-dimer equilibrium at pH 7.5 and in a monomer-tetramer equilibrium at pH 5.5. The pH appears monomer-tetramer equilibrium at pH 5.5. The pH appears to be a necessary and sufficient factor determining the types of oligomers formed. Although Ca2+ did not change the type of oligomerization, it had a very significant effect on the values of the thermodynamic parameters characterizing the associations. The delta G0 values for a monomer-dimer equilibrium were -7 to -8 kcal/mol, while those for a monomer-tetramer equilibrium were -20 to -23 kcal/mol. At pH 5.5, the values of delta H0, delta S0, and delta C0p were large and negative in the absence of Ca2+ and large and positive in the presence of 35 mM Ca2+, implying markedly different reaction mechanisms. Extrapolation of the results to 37 degrees C and 1 mM chromogranin A suggests that chromogranin A is virtually 100% tetramer at pH 5.5 in the presence of 35 mM Ca2+ but is 96% dimer at pH 7.5 in the absence of Ca2+, the two conditions resembling those seen in vivo. These results suggest that chromogranin A is mostly dimer in the endoplasmic reticulum and cis-Golgi area and is essentially all tetramer in the vesicle.

Of all the intracellular organelles, secretory granules contain by far the highest calcium concentration; secretory granules of typical neuroendocrine chromaffin cells contain approximately 40 mM Ca(2+) and occupy approximately 20% cell volume, accounting for >60% of total cellular calcium. They also contain the majority of cellular inositol 1,4,5-trisphosphate receptors (IP(3)Rs) in addition to the presence of >2 mM of chromogranins A and B that function as high-capacity, low-affinity Ca(2+) storage proteins. Chromogranins A and B also interact with the IP(3)Rs and activate the IP(3)R/Ca(2+) channels. In experiments with both neuroendocrine PC12 and nonneuroendocrine NIH3T3 cells, in which the number of secretory granules present was changed by either suppression or induction of secretory granule formation, secretory granules were demonstrated to account for >70% of the IP(3)-induced Ca(2+) releases in the cytoplasm. Moreover, the IP(3) sensitivity of secretory granule IP(3)R/Ca(2+) channels is at least approximately 6- to 7-fold more sensitive than those of the endoplasmic reticulum, thus enabling secretory granules to release Ca(2+) ahead of the endoplasmic reticulum. Further, there is a direct correlation between the number of secretory granules and the IP(3) sensitivity of cytoplasmic IP(3)R/Ca(2+) channels and the increased ratio of IP(3)-induced cytoplasmic Ca(2+) release, highlighting the importance of secretory granules in the IP(3)-dependent Ca(2+) signaling. Given that secretory granules are present in all secretory cells, these results presage critical roles of secretory granules in the control of cytoplasmic Ca(2+) concentrations in other secretory cells.-Yoo, S. H. Secretory granules in inositol 1,4,5-trisphosphate-dependent Ca(2+) signaling in the cytoplasm of neuroendocrine cells.

Human teeth system is one of the most important organs, plays vital role in mastication process. It consist 3 layers, outer hard and brittle cover enamel and porous dentine separated by very thin interfacial region called Dentine Enamel Junction (DEJ) or crown dentine. Dentine and enamel has almost isotropic material property while DEJ has different microstructure near enamel and dentine represented by functionally graded microstructure. Experimental evidences show that the crack initiated in the hard enamel region grows towards the crown dentine, it starts healing after 10–15 micron. Finite element analysis has been carried out and fracture toughness has been evaluated for layered model of human teeth with crack initiated at enamel region grows towards DEJ with its healing capabilities and graded structure. Comparison of the result with experimental value reported in literature shows acceptable matching suggests; FE results can be taken as basis for estimating Fracture toughness of human teeth system especially at interface of Enamel and Dentine, the DEJ.

Chromogranin A, the major intravesicular protein of adrenal chromaffin granules, bound Ca2+ in a pH-dependent manner. Both the maximal binding and affinity of chromogranin A for Ca2+ were dependent on pH. Chromogranin A bound 670 nmol of Ca2+/mg (32 mol/mol) and 1150 nmol of Ca2+/mg (55 mol/mol) at pH 7.5 and 5.5, respectively, with dissociation constants (Kd) of 2.7 and 4 mM. This pH dependence probably reflects different conformations of the protein at the two pH values. Conformational differences of chromogranin A at two different pH values were demonstrated by limited tryptic digestion patterns confirming previous results obtained by circular dichroism spectroscopy (Yoo, S. H., and Albanesi, J. P. (1990) J. Biol. Chem. 265, 14414-14421). Sedimentation equilibrium studies revealed the native molecular mass of chromogranin A to be 100 kDa at pH 7.5 and 192 kDa at pH 5.5, indicating dimeric and tetrameric states of the protein at the two pH levels. We postulate that the pH- and Ca2(+)-induced conformational changes of chromogranin A may have a role both in the regulation of Ca2+ release of chromaffin granules and in the early stages of secretory vesicle biogenesis.

The HSR-350x (high-speed rail 350x), also known as Korean G-7, is the experimental high speed train which was developed independently by G-7 R&D Project. The G-7 is now being investigated for its performance and tested at the high speed line. Partial side wear occurred in proposed braking system more than in KTX (Korean Train Express) according to experiment. In this study, comparison between KTX, KTX-II and G7 models of caliper unit of brake system has been done using FEM. The factor of generated partial side wear was investigated by comparing with results. Suitability of the caliper unit with KTX-II would be verified.

An investigation was performed to predict residual stress redistribution for the crack propagation initially through tensile residual stress field. The analytical method to estimate the redistribution of residual stress caused by crack propagation was proposed, on the basis of Dugdale model, by finite element analysis using elastic analysis method considering superposition principle. It is verified that the residual stress redistribution caused by crack propagation depends on configuration of the initial residual stress distribution. The variation aspect of redistribution of residual stress caused by crack propagation was examined based on the equilibrium condition of residual stresses.

Since Boucher technique used pedicle screw for the fixation of the spine in 1959, various kinds of pedicle screws have been developed continuously. The treatments of degenerative spine disease include spinal fusion and fixation using spinal devices. Spinal fusion surgery is classified into fusion and non fusion. The spine device market in which pedicle screws are used is getting bigger. However, it is one of the most challenging fields because of lack of the experimental and analytic data. In this study, a finite element model which includes non fusion device of human lumbar is developed and studied to understand the stability of the technique. The material property of the non fusion device is decided through comparison of intact model with fixed model. This result can be used as a basic source to new apparatus. This dynamic device offers a system that may alter the motion preservation and load sharing of a spinal segment without sacrificing stability.

The occlusal force of the tooth leads to loss of tooth tissue owing to attrition and abrasion, and may cause abfraction and pathological change of the dentine. Thus, we draw up finite element models, examine them applying ordinary occlusal force and analyze the stress distribution. Specimens used were mandibular first premolars from 15 Korean males and 13 females and were made into finite element models from medical images that were obtained using a micro-CT. We can find the fact that the irregular feature of the tooth is not useful only to masticating and pronouncing as well known, but that it is also suitable for protecting inner tissue by dispersing stress and delivering proper pressure to periodontal tissue to continue a physiological action. Also, image analysis can let us know the factor that is the cause of a disorder due to stress concentration in the cervical line. These results are expected to support the field of dental treatment planning, operating procedure and clinical trial, and the advance of technical expertise to develop implants and dentures.

Although the ciliate Tetrahymena is a good model for the study of chemotaxis, its profound motility makes it difficult to monitor intracellular calcium (Ca(2+)) changes induced by chemotactic stimuli. In this study, we report a microfluidic-based chemotaxis system generating directional chemotactic gradients under highly viscous conditions, suppressing T. pyriformis motility, and allowing for the stable confocal imaging of changes in intracellular Ca(2+) in the ciliate. Once the viscous condition was achieved, directional chemical gradients were formed inside the center chamber via the release of N-methyl-d-aspartate (NMDA), a known chemoattractant, from the surrounding chemical reservoirs into the center chamber. As a result, intracellular Ca(2+) in the ciliate increased up to three-fold, and its distribution was skewed in the direction of NMDA stimulation. However, the Ca(2+) in ciliates pretreated with phospholipase C (PLC) or phosphatidylinositol-3-kinase (PI3K) blockers did not increase even after stimulation. Additionally, the PI3K blocker induced the secretion of granules, the size of which was dependent on the concentration of the blocker. Collectively, the results indicate that both PLC and PI3K perform pivotal roles in controlling the levels of intracellular Ca(2+) in T. pyriformis during chemotaxis.

Inositol 1,4,5-trisphosphate receptor (IP(3)R) is a major intracellular Ca(2+) channel, modulated by many factors in the cytosolic and lumenal compartments. Compared with cytosolic control, lumenal-side regulation has been much less studied, and some of its mechanistic aspects have been controversial. Of particular interest with regard to lumenal regulation are whether it involves direct interactions between IP(3)R and the regulators, and whether it involves conformational changes of the lumenal regions of IP(3)R. To understand these lumenal-side regulation mechanisms, we studied the effects of two important lumenal regulatory factors, the redox potential and Ca(2+), on the L3-1 lumenal loop region of IP(3)R. The redox potential exerted direct and significant effects on the conformation of the loop region. By sharp contrast, Ca(2+) showed little effect on the L3-1 conformation, suggesting that the regulation of Ca(2+) is indirect or involves other receptor regions. GSH/oxidized glutathione-mediated oxidation introduced a unique intramolecular disulfide bond between Cys(34) and Cys(42). A variety of NMR experiments revealed that oxidation also induces localized helical characteristics in the Cys(34)-Cys(42) region. Dynamics studies also showed reduced motions in the region upon oxidation, consistent with the conformational changes. The results raise the interesting possibility that Cys(34) and Cys(42) may act together as a reduction sensor, and that Cys(65) may function as an oxidation sensor. Overall, our studies suggest that the redox potential and Ca(2+) can regulate IP(3)R through totally different mechanisms: Ca(2+) by the indirect effect and the redox potential by direct action causing conformational changes.

In this paper, the trend of FEM which is one of core tools of CAE and the future of CAE are examined in terms of TRIZ concept. This observation leads to the necessity of engineering creativity. This paper reports our successful experiences of implementing systematic innovation tools into engineering design classes. Also potential revolutionary change of engineering education is hinted by extending the accomplishments of TRIZ. It can innovate the current engineering curricula and an on-going study of classifying the engineering activities is introduced.

Chromogranin A, the most abundant protein in bovine adrenal chromaffin granules, bound calmodulin in a Ca2(+)-dependent manner, and the calmodulin-binding property was utilized to purify chromogranin A. Chromogranin A has been described in the past as a "random-coil polypeptide" with little alpha-helix or beta-sheet conformation. However, circular dichroism measurements with pure, native chromogranin A revealed relatively high alpha-helical contents (40% at the intravesicular pH of 5.5). Fluorescence studies confirmed previous observations that chromogranin A binds Ca2+ with low affinity. Considering the high concentration of Ca2+ in the secretory vesicle, the effect of Ca2+ on the secondary structure and self-association of chromogranin A was examined. Ca2+ induced a decrease of alpha-helicity of chromogranin A from 40 to 30% at pH 5.5. In contrast, at pH 7.5 the same amount of Ca2+ increased alpha-helicity of the protein from 25 to 40%. Boiling of the adrenal extract, a commonly used purification procedure for chromogranin A, resulted in the isolation of conformationally distinct chromogranin A molecule. Unlike secretory protein-I of the parathyroid gland (Gorr, S.-V., Dean, W. L., Radley, T. L., and Cohn, D. V. (1988) Bone Mineral 4, 17-25), chromogranin A aggregated rapidly in the presence of Ca2+. The extent and rate of aggregation were highly dependent on Ca2+ concentration. Although both the rate and extent of aggregation at pH 7.5 were much lower than those at pH 5.5, aggregation of chromogranin A proceeded at both pH's. In this respect, chromogranin A differs from human chromogranin C which was shown by Gerdes et al. (Gerdes, H.-H., Rosa, P., Phillips, E., Baeuerle, P. A., Frank, R., Argos, P., and Huttner, W. B. (1989) J. Biol. Chem. 264, 12009-12015) to aggregate at pH 5.2 but not at pH 7.4.

The inositol 1,4,5-trisphosphate receptor (IP3R) is a membrane channel that conducts calcium ions from the intracellular calcium stores. Despite a wealth of information on the cytoplasmic regulation of the IP3R, little is known about its regulation on the luminal side of the calcium stores. Here, we report studies on the IP3R intraluminal loop L3-2 and a conserved N-terminal region of chromogranin B. The IP3R loop is an important part of the channel's pore-forming region, and the chromogranin peptide has been shown to competitively inhibit calcium signaling by IP3R. Using the NMR titration approach, we showed that a part of the L3-2 is involved in a specific interaction with the chromogranin B peptide. Further NMR resonance assignments revealed that the 14th-20th residues of L3-2 are the keys to the binding to the chromogranin B peptide. Through detailed analysis of the data, we suggest a mechanism of IP3R regulation by chromogranin B involving conformational exchanges of the L3-2 region. Our report presents the findings of the first study on the interaction between the luminal loop of the IP3 receptor and its regulator at residue-resolution. The approaches described here should help to guide further studies on the interactions between the IP3R and other luminal side regulators.

Recently, secretory granule Ca(2+) storage protein chromogranin B (CGB) was shown to be present in the nucleoplasm proper in a complex structure that consists of the inositol 1,4,5-trisphosphate receptor (IP(3)R)/Ca(2+) channels and the phospholipids. Further, the amounts of IP(3)Rs present in the nucleus of bovine chromaffin cells were shown to be comparable to that of the endoplasmic reticulum. Therefore, we investigated here the potential contribution of nuclear CGB on the IP(3)-dependent Ca(2+) mobilization in the nucleus, using both neuroendocrine PC12 and nonneuroendocrine NIH3T3 cells. Chromogranin A (CGA) expression in the NIH3T3 cells, which do not contain intrinsic chromogranins, increased the IP(3)-induced Ca(2+) releases in the nucleus by 45%, while CGB expression in the same cells increased the IP(3)-induced Ca(2+) releases in the nucleus by 80%. Microinjection of IP(3) into the nucleus of CGB-expressing NIH3T3 cells increased the IP(3)-dependent nuclear Ca(2+) mobilization approximately 3-fold, whereas in CGA-expressing cells it remained the same as that of control cells. In contrast, inhibition of CGA expression in PC12 cells by siRNA treatment decreased the IP(3)-induced Ca(2+) releases in the nucleus by 17%, while inhibition of CGB expression decreased the IP(3)-induced Ca(2+) releases in the nucleus by 55%. Microinjection of IP(3) into the nucleus of siCGB-treated PC12 cells decreased the IP(3)-dependent nuclear Ca(2+) mobilization by approximately 75%, whereas in siCGA-treated cells it remained the same as that of control cells. Given the presence of CGB in the nucleus, these results further highlight the critical contribution of nuclear CGB in the IP(3)-induced Ca(2+) release in the nucleus.

We show here that expression of chromogranins in non-neuroendocrine NIH3T3 cells significantly increased the amount of IP 3 -mediated intracellular Ca 2 + mobilization in these cells, whereas suppression of them in neuroendocrine PC12 cells decreased the amount of mobilized Ca 2 + . We have therefore investigated the relationship between the IP 3 -induced intracellular Ca 2 + mobilization and secretory granules. The level of IP 3 -mediated Ca 2 + release in CGA-expressing NIH3T3 cells was 40% higher than in the control cells, while that of CGB-expressing cells was 134% higher, reflecting the number of secretory granules formed. Suppression of CGA and CGB expression in PC12 cells resulted in 41 and 78% reductions in the number of secretory granules, respectively, while the extents of IP 3 -induced Ca 2 + release in these cells were reduced 40 and 69%, respectively. The newly formed secretory granules of NIH3T3 cells contained all three isoforms of the IP 3 Rs. Comparison of the concentrations of the IP 3 R isoforms expressed in the ER and nucleus of chromogranin-expressing and nonexpressing NIH3T3 cells did not show significant differences, indicating that chromogranin expression did not affect the expression of endogenous IP 3 Rs. Nonetheless, the IP 3 R concentrations in secretory granules of chromogranin-expressing NIH3T3 cells were 3.5-4.7-fold higher than those of the ER, similar to the levels found in secretory granules of neuroendocrine chromaffin cells, thus suggesting that the IP 3 Rs targeted to the newly formed secretory granules are newly induced by chromogranins without affecting the expression of intrinsic IP 3 Rs. These results strongly suggest that the extent of IP 3 -induced intracellular Ca 2 + mobilization in secretory cells is closely related to the number of secretory granules.

The two major proteins of secretory granules of secretory cells, chromogranins A (CGA) and B (CGB), have previously been proposed to play key roles in secretory granule biogenesis. Recently, CGA was reported to play an on/off switch role for secretory granule biogenesis. In the present study we found CGB being more effective than CGA in inducing secretory granule formation in non-neuroendocrine NIH3T3 and COS-7 cells. The mean number of dense core granules formed/cell of CGA-transfected NIH3T3 cells was 2.51, whereas that of CGB-transfected cells was 4.02, indicating the formation of 60% more granules in the CGB-transfected cells. Similarly, there were 55% more dense core granules formed in the CGB-transfected COS-7 cells than in the CGA-transfected cells. Moreover, transfection of CGA- and CGB-short interfering RNA (siRNA) into neuroendocrine PC12 cells not only decreased the amount of CGA and CGB expressed but also reduced the number of secretory granules by 41 and 78%, respectively, further suggesting the importance of CGB expression in secretory granule formation.

Chromogranins A (CGA) and B (CGB) are two major Ca(2+) storage proteins of the secretory granules of neuroendocrine cells. Nevertheless, we found in the present study that CGB was also localized in the nucleus. In immunogold electron microscopy using bovine adrenal medullary chromaffin cells, it was found that the number of CGB-labeled gold particles localized per microm(2) of the nucleus was equivalent to 20% that of CGB-labeled gold particles localized per microm(2) of the secretory granules. Considering that CGB is estimated to exist in the 0.1-0.2-mm range in the secretory granules of bovine chromaffin cells, 20% of these amounts to 20-40 microm. In addition, transfection of CGA and CGB into nonneuroendocrine COS-7 and NIH3T3 cells repeatedly indicated the nuclear localization of CGB in addition to its usual localization in the cytoplasm. Moreover, immunoblot and immunogold electron microscopy analyses of neuroendocrine PC12 cells also showed the existence of endogenous CGB in both the cytosol and the nucleus. Nuclear routing of CGB did not appear to depend entirely upon the nuclear localization signal as some of the nuclear localization signal mutant CGB were still targeted to the nucleus. In gene array assay, CGB was shown to either induce or suppress transcription of many genes including those of transcription factors. Of these we have analyzed eight genes, four induced (zinc finger protein, MEF2C, hCRP2, abLIM) and four suppressed (hcKrox, T3-receptor, troponin C, integrin) using the quantitative reverse transcription-PCR method and spectrophotometry to determine the transcription levels of each mRNA. CGB was shown to increase the transcription of zinc finger protein, MEF2C, hCRP2, and abLIM by 2.5-5-fold while suppressing that of hcKrox, T3-receptor, troponin C, and integrin by 60-75%. Given that MEF2C and hcKrox genes are transcription factors, these results pointed to the transcription control role of CGB in the nucleus.

Secretogranin II (SgII) is one of the three major proteins, the other two being chromogranins A (CGA) and B (CGB), of secretory granules of neuroendocrine cells. The Ca(2+) storage proteins CGA and CGB not only are coupled to the IP(3) receptor (IP(3)R)/Ca(2+) channels that exist on the secretory granule membrane but also are known to play key roles in secretory granule biogenesis. Unlike the better studied CGA and CGB, secretogranin II has never been completely purified in the native state and studied. We have therefore purified SgII in native form from bovine adrenal medulla and subjected it to biochemical characterization. Secretogranin II consisted of largely beta-sheet and random coil structures with a low level of alpha-helicity. Like CGA and CGB, it also underwent pH-dependent conformational changes, showing 9.5% alpha-helicity at pH 7.5 and 17.0% alpha-helicity at pH 5.5. Secretogranin II also underwent acidic pH- and Ca(2+)-dependent aggregation, and it was approximately 8-fold more sensitive than CGA to Ca(2+) in its pH-dependent aggregation but was 8-fold less sensitive than CGB. Further, similar to CGA and CGB that had interacted with the secretory granule membrane at the intragranular pH 5.5, SgII also interacted with the secretory granule membrane at pH 5.5 and dissociated from it at near-physiological pH 7.5, implying similar roles of SgII in the cell as those of CGA and CGB. Secretogranin II hence appeared to actively participate in secretory granule biogenesis as has been proposed for CGA and CGB.

It has been proposed that chromogranin A (CgA), a protein secreted by many normal and neoplastic neuroendocrine cells, can play a role as a positive or a negative modulator of cell adhesion. The mechanisms that regulate these extracellular functions of CgA are unknown. We show here that plasmin can regulate the anti/pro-adhesive activity of CgA by proteolytic cleavage of the N-terminal domain. Limited proteolytic processing decreased its anti-adhesive activity and induced pro-adhesive effects in fibronectin or serum-dependent fibroblast adhesion assays. Cleavage of Lys(77)-Lys(78) dibasic site in CgA(1-115) was relatively rapid and associated with an increase of pro-adhesive effect. In contrast, antibodies against the region 53-90 enhanced the anti-adhesive activity of CgA and CgA(1-115). Structure-activity relationship studies showed that the conserved region 47-64 (RILSILRHQNLLKELQDL) is critical for both pro- and anti-adhesive activity. These findings suggest that CgA might work on one hand as a negative modulator of cell adhesion and on the other hand as a precursor of positive modulators, the latter requiring proteolytic processing for activation. Given the importance of plasminogen activation in tissue invasion and remodeling, the interplay between CgA and plasmin could provide a novel mechanism for regulating fibroblast adhesion and function in neuroendocrine tumors.

Partial strain relaxation effects on polarization ratio of semipolar (1122) InxGa1−xN/GaN quantum well (QW) structures grown on relaxed InGaN buffers were investigated using the multiband effective-mass theory. The absolute value of the polarization ratio gradually decreases with increasing In composition in InGaN buffer layer when the strain relaxation ratio (e0y′y′−ey′y′)/e0y′y′ along y′-axis is assumed to be linearly proportional to the difference of lattice constants between the well and the buffer layer. Also, it changes its sign for the QW structure grown on InGaN buffer layer with a relatively larger In composition (x > 0.07). These results are in good agreement with the experiment. This can be explained by the fact that, with increasing In composition in the InGaN subsrate, the spontaneous emission rate for the y′-polarization gradually increases while that for x′-polarization decreases due to the decrease in a matrix element at the band-edge (k‖ = 0).

중심성 기도 폐쇄는 양성 혹은 악성 질환과 동반되어 나타날 수 있으며 현재 다양한 기관지경적 치료법이 사용되고 있다. IT knife-2는 소화기 내시경적 시술을 위해 고안된 장비로서 그 안정성과 효용성이 입증되어 국내 의료기관에서 내시경적 점막하 박리술에 널리 사용되고 있다. 저자들은 굴곡성 기관지경과 IT knife-2를 이용하여 진행성 폐암과 방사선 치료 후 발생한 중심성 기도 폐쇄를 가진 두 환자를 성공적으로 치료하였다. 합병증과 예후에 대한 추가적인 연구가 필수적이지만 IT knife-2는 향후 기도 폐쇄의 내시경적 중재적 시술법 중 하나로 고려될 수 있을 것이다.

Pulmonary tumor thrombotic microangiopathy (PTTM) is known as a rare and severe cancer-related pulmonary complication that was first reported by Von Herbay et al. in 1990 [1]. In PTTM cases, endothelial attachment of multiple microscopic tumor emboli induces intimal proliferation of small pulmonary arteries and arterioles combined with secondary thrombosis, which leads to massive reduction of the pulmonary vascular bed and pulmonary hypertension [1-3]. Due to extremely rapid progression of PTTM and non-specific clinical findings, establishing a PTTM diagnosis is very difficult, and nearly all reported cases are diagnosed primarily by autopsy [1,3]. There have only been two PTTM cases reported in Korea, but they were diagnosed by autopsy or shortly before death, and thus any effective treatment was not available [4,5]. There have been only three cases worldwide that have been diagnosed ante mortem and treated effectively [6-8]. We report a 44-year-old woman who did not have a history of cancer but presented with rapidly progressive dyspnea and pulmonary hypertension. She was diagnosed as advanced gastric cancer and related PTTM ante mortem. She was treated effectively with chemotherapy for the underlying malignancy.

A simple parabolic formula that describes the opening profile of a sufficiently long circumferential crack in a pipe subjected to bending and tension was found after a proper normalization of the second-order asymptotic expansion of the solutions of Sanders and Alabi [5-9]. Here the crack is sufficiently long in the sense that approximately the crack angle is larger than a small parameter which has the same order of magnitude as the ratio of the wall thickness to the pipe radius, and the pipe is sufficiently long enough not to consider the end effects. These linear elastic solutions can be superposed for combined bending and tension loading. The smile parabolic formula is shown to be a good approximation of the crack opening profile and can be used to evaluate the leak rate for crack length up to one-half of the circumference.