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The irritant action of capsaicin is mediated by the vanilloid receptor, VR1, which is expressed in sensory neurons termed nociceptors. Capsaicin also desensitizes nociceptors and, thus, is useful clinically as an analgesic. Given the potential importance of VR1 in pain, we have cloned the human capsaicin receptor, hVR1, from a human dorsal root ganglia (DRG) cDNA library. Human VR1 protein is 85% identical to the rat VR1 and many of the amino acid differences are concentrated at the amino and carboxyl termini. VR1 is expressed in DRG as an approximately 4.2 kilobase RNA, and is also expressed in the central nervous system and in the kidney. Capsaicin (EC(50) = 853 nM), low pH (

Hepatitis C virus (HCV) infection is a leading cause of liver-related mortality. Chronic hepatitis C (CHC) is frequently associated with disturbances in iron homeostasis, with serum iron and hepatic iron stores being elevated. Accumulating evidence indicates that chronic HCV infection suppresses expression of hepatic hepcidin, a key mediator of iron homeostasis, leading to iron overload conditions. Since hepcidin mediates degradation of ferroportin, a basolateral transporter involved in the release of iron from cells, diminished hepcidin expression probably leads to up-regulation of ferroportin-1 (Fpn1) in patients with CHC. In this study, we determined the protein levels of duodenal Fpn1, and found that its expression was significantly up-regulated in patients with CHC. The expression of duodenal Fpn1 is negatively correlated with mRNA levels of hepcidin, and positively correlated with serum iron parameters. Although iron is a critical factor for growth of a variety of pathogenic bacteria, our results suggest that iron overload in blood does not increase the infection rate of bacteria in patients with CHC.

The consequences of long-term in vivo expression of human c-mpl ligand in a mouse model were examined. Transgenic mice expressing the human full-length cDNA in the liver exhibited a fourfold increase in circulating platelet count that persisted stably over the life of the animals. Transgenic animals thrived and appeared healthy for at least 500 days. Transgenic platelets appeared normal with respect to surface antigens and response to platelet aggregation agonists. The highest-expressing transgenic line maintained human c-mpl ligand serum levels of 3 ng/mL. Megakaryocyte numbers in bone marrow and spleen were elevated, as were bone marrow and spleen megakaryocyte colony-forming cells (MEG-CFC). Megakaryocytes were observed in the bone marrow, spleen, liver, and lung, but in no other sites. Circulating myeloid and lymphoid cell populations were increased twofold. Additionally, the animals had a slight but significant anemia despite an increase in marrow colony-forming units-erythroid (CFU-E). No evidence of myelofibrosis was observed in the bone marrow. The platelet nadir in response to administration of either antiplatelet serum (APS) or 5-fluorouracil (5FU) was significantly reduced relative to the control level. Furthermore, the red blood cell (RBC) nadir was reduced relative to control levels in both models, suggesting that c-mpl ligand can directly or indirectly support the maintenance of erythrocyte levels following thrombopoietic insult.

Mutations within microsatellite sequences, consisting of additions or deletions of repeat units, are known as the replication/repair error positive (RER+) phenotype or micorsatellite instability (MI). Microsatellite instability has been demonstrated in hereditary and sporadic colorectal carcinomas and is usually observed in noncoding regions of genomic DNA. However, relatively few coding region targets of MI have been identified thus far. Using PCR, we amplified regions encompassing (A)8 and (C)8 microsatellite tracts within hMSH3 and hMSH6 from 31 RER+ sporadic colorectal tumors, 8 hereditary colon cancers, 23 RER+ gastric carcinomas, and 32 RER- gastric tumors. Mutations were found in 11 (36%) of 31 sporadic colon carcinomas, 4 (50%) of 8 hereditary colorectal cancers, and 5 (22%) of 23 RER+ gastric carcinomas, but in only 2 (6%) of 32 RER- gastric carcinomas. These frameshift mutations cause premature stop codons downstream that are predicted to abolish normal protein function. Our results and those of others suggest that DNA mismatch repair genes, such as hMSH3 and hMSH6, are targets for the mutagenic activity of upstream mismatch repair gene mutations and that this enhanced genomic instability may accelerate the accumulation of mutations in RER+ tumors.

An inspection system for indium tin oxide (ITO) circuits has been developed with the use of a polymer dispersed liquid crystal (PDLC)/ITO film which was used as a sensing device to locate faulty shut/open circuits. With the power on, the PDLC/ITO film covering the conducting area was changed from translucent state to clear state while those covering the non-conducting area was remain translucent. In this study, simulations and experiments were conducted to evaluate the performance of the proposed system and study effects of system parameters on the limitation of the proposed system. It is of interest to find that the increase in applied external voltage will improve the capability of identifying conducting area while that will generally degrade the capability of identifying non-conducting area with the range studied.

In this study, it is of interest to find a way to inspect transparent circuits with narrow line widths (

Unmanned aerial vehicle (UAV) had developed for many years, with quad-rotor vertical take-off and landing (VTOL) unmanned aerial vehicle developing fastest. The equations of its flight attitude have been derived and verified by some scholars. Up to now, the quad-rotor unmanned aerial vehicle has already become one of the unmanned aerial vehicles (UAV) that can fly stably. Today, when the robot technology is booming, various sensors and intelligent functions that applied in robots can be transferred and applied to the quad-rotor unmanned aerial robot. It will make the quad-rotor unmanned aerial robot not an ordinary UAV any longer, but an intelligent robot that can fly. In this research, the image processing technique had been integrated with the quad-rotor aerial robot. The aerial robot cannot receive the GPS signal for positioning in an indoor environment. With the image processing technique, the quadcopter can hover around a fixed position or visual track a moving object in an indoor environment.

Fibroblast growth factor receptor 4 (FGFR4) plays crucial roles in vascular smooth muscle cell proliferation and atherosclerosis and, therefore, may potentially affect the development of coronary artery disease (CAD). The aim of this study was to investigate the association between FGFR4 polymorphisms and the susceptibility to CAD in the Chinese population. Two polymorphisms, rs192201146G/A (Asp756Asn) and rs188755817C/G (Ser778Arg), were detected by polymerase chain reaction-restriction fragment length polymorphism and direct sequencing in 722 CAD cases and 802 age-matched controls. Data were analyzed using the chi-square test. Results showed that frequencies of rs192201146GA genotype and rs188755817CG genotype were significantly higher in CAD patients than in controls (odds ratio [OR]=1.92, 95% confidence interval [CI] 1.11-3.28, p=0.016, and OR=1.87, 95% CI 1.06-3.30, p=0.027). Similarly, numbers of the rs192201146A allele and the rs188755817G allele were significantly increased in CAD cases (OR=1.89, 95% CI 1.11-3.22, p=0.017, and OR=1.85, 95% CI 1.06-3.24, p=0.029). Haplotype analysis revealed that GG and AC (rs192201146 rs188755817) haplotypes had higher frequencies in CAD patients (OR=2.75, p=0.002 and OR=2.69, p=0.001). Our data suggested that the FGFR4 rs192201146 (Asp756Asn) and rs188755817 (Ser778Arg) polymorphisms could act as risk factors for CAD in the Chinese population.

In the last few years, intelligent robots were successfully fielded in hospitals (King, S., and Weiman, C, 1990), museums (Burgard, W. et al., 1999), and office buildings/department stores (Endres, H. et al. , 1998), where they perform cleaning services, deliver, educate, or entertain (Schraft, R., and Schmierer, G. , 2005). Robots have also been developed for guiding blind people, as well as robotic aids for the elderly. Today, the number of elderly in need of care is increasing dramatically. As the baby-boomer generation approaches the retirement age, this number will increase significantly. Current living conditions for the majority of elderly people are already unsatisfactory, and situation will worsen in the future. (Nicholas Roy et al. , 2000) Rapid progress of standard of living and health care resulted in the increase of aging population. More and more elderly people do not receive good care from their family or caregivers. Maybe the intelligent service robots can assist people in their daily living activities. Robotics aids for the elderly have been developed, but many of these robotics aids are mechanical aids. (Song, W.-K. et al., 1998) (Dario, P. et al., 1999) (Takahashi, Y. et al., 1999). The intelligent service robot can assist elderly people with many tasks, such as remembering to take medicine or measure blood pressure on time. The main objective of this Chapter is to develop an omnidirectional mobile home care robot. This service mobile robot is equipped with “Indoor positioning system”. The indoor positioning system is used for rapid and precise positioning and guidance of the mobile robot. Five reflective infrared sensors are placed around the robot for obstacle avoidance. The wireless IP camera is placed on the top layer of this robot. Through the internet remote control system, the live image of the IP camera on the robot can be transferred to the remote client user. With this internet remote control system, the remote client user can monitor the elderly people or the home security condition. On the aid of this system, remote family member can control the robot and talk to the elderly. This intelligent robot also can deliver the medicine or remind to measure the blood pressure or blood sugar on time. We hope this intelligent robot can be a housekeeper or family guard to protect our elderly people or our family. The functions of the proposed robot are illustrated as follows:  deliver medicine or food on time  remind to measure and record the blood pressure or blood sugar of the elderly on time

Hyperhomocysteinemia (HHcy) has been shown to induce endothelial dysfunction, an early event in the progression of atherosclerosis. However, the underlying mechanism of endothelial cell injury in HHcy has not been clearly elucidated. In this study, we examined the effect of homocysteine on tribbles-related protein 3 (TRB3)-mediated cell-cycle arrest in human umbilical vein endothelial cells (HUVECs). Treatment of HUVECs with homocysteine (0–250 µmol/L) resulted in inhibition of cell proliferation assessed by [3H]-thymidine incorporation into DNA. Homocysteine induced cell-cycle arrest in the G1 phase by up-regulating the protein levels of p27kip1. Under these conditions, homocysteine did not induce endoplasmic reticulum stress. However, homocysteine up-regulated the expression of TRB3, thus leading to the dephosphorylation of Akt (Thr308). Knock-down of endogenous TRB3 using siRNA significantly suppressed the inhibitory effect of homocysteine on the proliferation of HUVECs. Homocysteine-induced TRB3 expression was mediated by the cAMP/cAMP response element-binding protein (CREB) pathway. These results demonstrate that TRB3 is a critical molecule in the homocysteine-mediated cell-cycle arrest in endothelial cells. J. Cell. Physiol. 226: 2782–2789, 2011. © 2011 Wiley-Liss, Inc.

Phase unwrapping is a very important processing step in phase shift interferometry. In this work, we propose a new method which combines the branch-cut method with error correcting. The method can avoid the propagation of the phase errors and have higher reliability. The experiment proves the proposed method is feasible and effective.

To investigate the role of reactive oxygen species (ROS) and the effect of vitamin E on proliferation of vascular smooth muscle cells (VSMCs) induced by homocysteine.DNA synthesis in the VSMCs cells was measured using [3H]-thymidine incorporation assay, and the cell number determined by trypan blue method. The level of ROS in the cells was determined using DCF-DA as the fluorescence probe.Homocysteine promoted VSMC DNA synthesis, proliferation, and ROS production. Cysteine resulted in increased ROS production in VSMCs, but had no significant effect on DNA synthesis and cell proliferation. Catalase significantly inhibited ROS production induced by homocysteine, but did not significantly inhibited homocysteine-mediated proliferation of VSMCs. While alpha-tocopherol and beta-tocopherol both suppressed increased ROS production induced by homocysteine in VSMCs, only alpha-tocopherol significantly inhibited homocysteine-mediated VSMC proliferation.ROS is not associated with VSMC proliferation, and vitamin E-induced suppression of VSMC proliferation is probably related to protein kinase C inhibition.

To investigate the effect of negative charge aerosol (NCA) on the treatment of burn wound.Patients with superficial or deep partial thickness burn only were enrolled in the study, and they were randomly divided into trial group (T, including 180 cases of superficial thickness burn and 100 cases of deep partial thickness burn), control group (C, including 30 cases with superficial thickness burn and 30 with deep partial thickness burn), and self control group (SC, including 10 cases with superficial thickness burn and 10 with deep partial thickness burn). The patients in T and SC groups were treated with NCA for 1.5 hours, 1-2 times a day, from 6 postburn hour (PBH) to 2 postburn day (PBD), while those in C group received conventional treatment. For those in SC group, some of the wounds were covered with sterile schissel, while other wounds without schissel covering. The general changes in the wounds during NCA treatment were observed, and bacterial culture before and after NCA treatment was performed. The healing time was recorded and the blood biochemical parameters were determined. Rat model with deep partial thickness scald was established, and the rats were also divided into T and C groups, and received treatment as in human. Tissue samples were harvested from the wounds of rats in the 2 groups before and 1, 2, 3 weeks after treatment for pathological examination.There was no infection and little exudation in the patients in T group. No bacteria were found in the wound before and after NCA treatment. The healing time of the wounds of patients with superficial and deep partial thickness burn in T group was 6.3 +/- 1.6 d and 15.1 +/- 3.1 d, respectively, which was obviously shorter than those in C group (11.3 +/- 1.4 d and 21.2 +/- 1.4 d, P0.01). In SC group, the healing time of those with sterile schissel coverage was also significantly shorter than those without covering (P0.01). There was no obvious change in the liver and kidney functions and blood biochemical parameters among the patients. Pathological examination showed that the skin structure was almost recovered in the rats in T group 3 weeks after treatment, while those in C group was not.Negative charge aerosol is safe and effective in promoting wound healing of the patients with partial thickness burns.

This paper develops a multiobjective optimization methodology for automotive door quality design under uncertainty, in which the tradeoffs between two competing objectives need to be considered. Two important quality issues, door closing effort and wind noise, are of concern and the corresponding probabilities of unsatisfactory performance are considered in the optimization. Model-based reliability analysis methods are used to compute these probabilities. Both component and system-level reliability analyses need to be performed in RBDO. While a first order reliability method (FORM) is found adequate for the reliability estimation with respect to door closing effort, an adaptive Monte Carlo simulation method is found suitable for reliability analysis of the wind noise problem with multiple limit states. An efficient decoupled RBDO approach is used to solve the multiobjective optimization and the Pareto frontier is generated for decision-making. The proposed method can be applied to solve a wide variety of RBDO problems with competing objectives and reliability constraints at both component and system levels.

Highly oriented ferroelectric strontium barium niobate (Sr 0.6 Ba 0.4 Nb 2 O 6 ) thin films were prepared on P-type Si(100) substrate by the Sol-Gel process. The XRD patterns of the SBN films show that SBN film prepared by using NbCl5, KOH as raw materials performed a highly c-axis preferred orientation perpendicular to the Si substrate, better than films that was prepared using Nb(OC 2 H 5 ) 5 as starting agents. It may be duo to the existence of the potassium ion that not be filtered out completely during the preparation of the niobium alkoxide. The characteristics of D-F and C-V curves were obtained for SBN/Si film. The film exhibits high dielectric constant. In order to investigate ferroelectric characteristics further, the P-E loops of the SBN/Pt/Si were also measured. The films show better optical properties, transmittance of Sr 0.6 Ba 0.4 Nb 2 O 6 films on MgO(001) and SiO 2 substrates was more than 60% at the range from 450 to 850nm, refractive index was measured to be 2.14 and 2.12 on the MgO and SiO 2 substrate at 633nm respectively.

ZnO were deposited on sapphire and silicon substrates by RF magnetron sputtering from a metallic zinc target. The structural and optical propertied of ZnO films were studied by X-ray diffraction, and UV-VIS-NIR scanning spectrophotometer. XRD measurements show ZnO films had a preferential orientation along the c-axis. Only one peak, (002) phase, appears on the diffraction spectra. The transmittance spectra indicate that ZnO films possessed a transmittance of about 80% in the visible region and a sharp absorption edge at wavelength of about 390nm. The refractive index n and the extinction coefficient k are all sensitive to the oxygen partial pressure and the substrate temperature. Furthermore, based on the ideal five layers symmetrical waveguide films, the relationships of the loss and the thickness of the waveguide layer and the buffer layer were analyzed using ZnO as waveguide layer and SiO 2 as buffer layer.

A new color interpolation method is proposed in this paper. We use the correlation of R, G, and B pixel, 'Lagrange Interpolation', and edge indicator to interpolate the DSC of single CCD.

A multiobjective optimization methodology under uncertainty is developed. Model-based reliability analysis methods are used to compute the probabilistic constraints. Both component-level and system-level reliability analyses are included in the optimization formulation. Several approaches in multiobjective optimization are used to generate the Pareto frontier for decisionmaking. The proposed method is successfully applied to a car door subsystem, in which two important quality issues, door closing effort and wind noise, are of concern and a balanced design needs to be achieved.

An accurate and efficient Monte Carlo simulation (MCS) method is developed in this paper for limit state-based reliability analysis, especially at system levels, by using a response surface approximation of the failure indicator function. The Moving Least Squares (MLS) method is used to construct the response surface of the indicator function, along with an Optimum Symmetric Latin Hypercube (OSLH) as the sampling technique. Similar to MCS, the proposed method can easily handle implicit, highly nonlinear limit-state functions, with variables of any statistical distributions and correlations. However, the efficiency of MCS can be greatly improved. The method appears to be particularly efficient for multiple limit state and multiple design point problem. A mathematical example and a practical example are used to highlight the superior accuracy and efficiency of the proposed method over traditional reliability methods.

A novel reliability-based design optimization (RBDO) method using simulation-based techniques for reliability assessments and efficient optimization approach is presented in this paper. In RBDO, model-based reliability analysis needs to be performed to calculate the probability of not satisfying a reliability constraint and the gradient of this probability with respect to each design variable. Among model-based methods, the most widely used in RBDO is the first-order reliability method (FORM). However, FORM could be inaccurate for nonlinear problems and is not applicable for system reliability problems. This paper develops an efficient optimization methodology to perform RBDO using simulation-based techniques. By combining analytical and simulation-based reliability methods, accurate probability of failure and sensitivity information is obtained. The use of simulation also enables both component and system-level reliabilities to be included in RBDO formulation. Instead of using a traditional RBDO formulation in which optimization and reliability computations are nested, a sequential approach is developed to greatly reduce the computational cost. The efficiency of the proposed RBDO approach is enhanced by using a multi-modal adaptive importance sampling technique for simulation-based reliability assessment; and by treating the inactive reliability constraints properly in optimization. A vehicle side impact problem is used to demonstrate the capabilities of the proposed method.Copyright © 2004 by ASME

In this paper, an efficient method for the reliability analysis of systems with nonlinear limit states is described. It combines optimization-based and simulation-based approaches and is particularly applicable for problems with highly nonlinear and implicit limit state functions, which are difficult to solve by conventional reliability methods. The proposed method consists of two major parts. In the first part, an optimization-based method is used to search for the most probable point (MPP) on the limit state. This is achieved by using adaptive response surface approximations. In the second part, a multi-modal adaptive importance sampling method is proposed using the MPP information from the first part as the starting point. The proposed method is applied to the reliability estimation of a vehicle body-door subsystem with respect to one of the important quality issues -- the door closing effort. The superiority of the proposed method, in terms of efficiency and accuracy, is demonstrated with a numerical example of highly nonlinear limit state problem, as well as an automotive door closing effort application. A generalized framework for reliability estimation is also proposed for problems with large numbers of random variables and complicated limit states.

Frequent loss of heterozygosity (LOH) on human chromosome 7q31 has been reported in numerous malignancies. Suppressor of tumorigenicity 7 (ST7) has been identified as a candidate tumor suppressor gene in this region. To identify whether 7q31 and genetic alterations of ST7 were involved in human esophageal carcinogenesis, we performed LOH mapping of a 5.4 cM region at 7q31-q35 in 43 primary esophageal carcinomas, as well as mutational analyses of the ST7 gene in tumors with LOH in this region. Of 43 tumors, 12 (28%) showed LOH at 7q31–q35. These included four (22%) of 18 squamous cell carcinomas and eight (32%) of 25 adenocarcinomas. The peak LOH locus was D7S480, lying 4.2 Mb telomeric to ST7 and showing LOH in eight of 37 informative tumors, or 22%. No mutations were found in the entire coding or flanking intronic regions of the ST7 gene among 12 tumors with 7q-LOH. In addition, quantitative RT–PCR analyses of ST7 mRNA expression levels in 11/13 normal-tumor pairs failed to show more than a 50% decrease in tumor ST7 mRNA relative to matched normal tissues. These data suggest that LOH at 7q31–q35 is involved in the origin or progression of at least a subset of esophageal carcinomas, but that ST7 is not the target gene of this somatic event.

Reliability analysis methods are commonly used in engineering design, in order to meet reliability and quality measures. An accurate and efficient computational method is presented for reliability analysis of engineering systems at both the component and system levels. The method can easily handle implicit, highly nonlinear limit-state functions, with correlated or non-correlated random variables, which are described by any probabilistic distribution. It is based on a constructed response surface of an indicator function, which determines the “failure” and “safe” regions, according to the performance function. A Monte Carlo simulation (MCS) calculates the probability of failure based on a response surface of the indicator function, instead of the computationally expensive limit-state function. The Cross-Validated Moving Least Squares (CVMLS) method is used to construct the response surface of the indicator function, based on an Optimum Symmetric Latin Hypercube (OSLH) sampling technique. A number of numerical examples highlight the superior accuracy and efficiency of the proposed method over commonly used reliability methods.Copyright © 2003 by ASME

Microsatellite instability (MSI) within coding regions causes frameshift mutations (FSMs). This type of mutation may inactivate tumor suppressor genes in cancers with frequent MSI (MSI-H cancers). To identify novel FSMs in gastric carcinogenesis in an unbiased and comprehensive manner, we screened for this type of mutation at 154 coding region repeat loci in 18 MSI-H gastric cancers. We also compared FSM rates and spectra in MSI-H gastric versus colorectal cancers. Thirteen novel loci showed FSMs in20% of gastric tumors. Novel loci with the highest mutation frequencies included the activin type 2 receptor gene (44.4%), DKFZp564K112 (a homologue of the Drosophila tumor suppressor gene multi-sex-combs; 41.2%), and an endoplasmic reticulum chaperone protein gene SEC63 (37.5%). The mutational spectra for genes with high mutation frequencies were also significantly different between MSI-H gastric and colorectal cancers.

cDNAmicroarrays, combined with bioinformatics analyses, are becomingincreasingly used in current medical research. Existing analytic methods,particularly those that are unsupervised, often have difficulty recognizing subtle differences among predefined subgroups. In contrast, supervised methods, such as Artificial Neural Networks (ANNs), are able to recognize subtly different biological entities. We applied ANNs in a proof-of-principle study of cDNA microarray data in esophageal cancer (CA) and premalignancy. cDNA microarrays, each containing 8064 clones, were hybridized to RNAs from 22 esophageal lesions, including 14 Barrett's esophagus (BA) metaplasias and 8 esophageal carcinomas (3 squamous cell carcinomas and 5 adenocarcinomas). Scanned cDNA microarray data were analyzed using the bioinformatics software Cluster/TreeView, Significance Analysis of Microarrays (SAM), and ANNs. Cluster analysis based on all 8064 clones on the microarrays was unable to correctly distinguish BA specimens from CA specimens. SAM then selected 160 differentially expressed genes between Barrett's and cancer. Cluster analysis based on this reduced set still misclassified 2 Barrett's as cancers. The ANN was trained on 12 samples and tested against the remaining 10 samples. Using the 160 selected genes, the ANN correctly diagnosed all 10 samples in the test set. Finally, the 160 genes selected by SAM may merit further study as biomarkers of neoplastic progression in the esophagus, as well as in elucidating pathological mechanisms underlying BA and CA.

Esophagin is a member of the small proline-rich protein family of cell envelope precursor proteins, which are expressed during squamous cell differentiation. Esophagin is expressed at high levels in normal esophageal epithelium, but its expression is absent from esophageal squamous cell carcinomas and adenocarcinomas. Moreover, loss of esophagin expression is present in areas of dysplasia or normal mucosa adjacent to carcinomas, suggesting that absence of esophagin may constitute a harbinger of early esophageal malignant transformation. A greater understanding of transcriptional control of esophagin may provide valuable insights into esophageal malignancy. Therefore, this study was undertaken in order to isolate and carry out initial characterization of a functional promoter for esophagin. A genomic clone containing esophagin was isolated and sequenced, including 2.7 kb of the esophagin promoter region. Esophagin expression was studied in response to various treatments of primary cultured human esophageal epithelial cells and squamous cell carcinoma cell lines. Calcium was the strongest inducer of the endogenous esophagin promoter, with induction occurring at 12–72 hours. In primary cultured esophageal epithelial cells, a region spanning 116 bp upstream of the transcriptional start site to 45 bp downstream was sufficient to direct low, basal, in vitro esophagin expression. However, responsiveness of primary esophageal cells to calcium required inclusion of promoter elements 1688 bp upstream of the transcriptional start site. Site-directed mutagenesis studies suggested a putative role for C/EBP-β, OCT-1, and OCT-3 transcription factor binding sites in the minimal promoter region. In conjunction with published human in vivo studies, these data support the hypothesis that esophagin is a biomarker of esophageal squamous cell differentiation and provide an in vitro model to evaluate regulatory factors involved in this differentiation process.

Background & Aims: There is a subtle distinction between sporadic colorectal adenomas and cancers (SAC) and inflammatory bowel disease (IBD)-associated dysplasias and cancers. However, this distinction is clinically important because sporadic adenomas are usually managed by polypectomy alone, whereas IBD-related high-grade dysplasias mandate subtotal colectomy. The current study evaluated the ability of artificial neural networks (ANNs) based on complementary DNA (cDNA) microarray data to discriminate between these 2 types of colorectal lesions. Methods: We hybridized cDNA microarrays, each containing 8064 cDNA clones, to RNAs derived from 39 colorectal neoplastic specimens. Hierarchical clustering was performed, and an ANN was constructed and trained on a set of 5 IBD-related dysplasia or cancer (IBDNs) and 22 SACs. Results: Hierarchical clustering based on all 8064 clones failed to correctly categorize the SACs and IBDNs. However, the ANN correctly diagnosed 12 of 12 blinded samples in a test set (3 IBDNs and 9 SACs). Furthermore, using an iterative process based on the computer programs GeneFinder, Cluster, and MATLAB, we reduced the number of clones used for diagnosis from 8064 to 97. Even with this reduced clone set, the ANN retained its capacity for correct diagnosis. Moreover, cluster analysis performed with these 97 clones now separated the 2 types of lesions. Conclusions: Our results suggest that ANNs have the potential to discriminate among subtly different clinical entities, such as IBDNs and SACs, as well as to identify gene subsets having the power to make these diagnostic distinctions.

In order to discover global gene expression patterns characterizing subgroups of colon cancer, microarrays were hybridized to labeled RNAs obtained from seventeen colonic specimens (nine carcinomas and eight normal samples). Using a hierarchical agglomerative method, the samples grouped naturally into two major clusters, in perfect concordance with pathological reports (colon cancer versus normal colon). Using a variant of the unpaired t-test, selected genes were ordered according to an index of importance. In order to confirm microarray data, we performed quantitative, real-time reverse transcriptase-polymerase chain reaction (TaqMan RT-PCR) on RNAs from 13 colorectal tumors and 13 normal tissues (seven of which were matched normal-tumor pairs). RT-PCR was performed on the gro1, B-factor, adlican, and endothelin converting enzyme-1 genes and confirmed microarray findings. Two hundred and fifty genes were identified, some of which were previously reported as being involved in colon cancer. We conclude that cDNA microarraying, combined with bioinformatics tools, can accurately classify colon specimens according to current histopathological taxonomy. Moreover, this technology holds promise of providing invaluable insight into specific gene roles in the development and progression of colon cancer. Our data suggests that a large-scale approach may be undertaken with the purpose of identifying biomarkers relevant to cancer progression.

A significant portion of gastric cancers exhibit defective DNA mismatch repair, manifested as microsatellite instability (MSI). High-frequency MSI (MSI-H) is associated with hypermethylation of the human mut-L homologue 1 (hMLH1) mismatch repair gene promoter and diminished hMLH1 expression in advanced gastric cancers. However, the relationship between MSI and hMLH1 hypermethylation has not been studied in early gastric neoplasms. We therefore investigated hMLH1 hypermethylation, hMLH1 expression and MSI in a group of early gastric cancers and gastric adenomas. Sixty-four early gastric neoplasms were evaluated, comprising 28 adenomas, 18 mucosal carcinomas, and 18 carcinomas with superficial submucosal invasion but clear margins. MSI was evaluated using multiplex fluorescent PCR to amplify loci D2S123, D5S346, D17S250, BAT 25 and BAT 26. Methylation-specific PCR was performed to determine the methylation status of hMLH1. In two hypermethylated MSI-H cancers, hMLH1 protein expression was also evaluated by immunohistochemistry. Six of sixty-four early gastric lesions were MSI-H, comprising 1 adenoma, 4 mucosal carcinomas, and 1 carcinoma with superficial submucosal invasion. Two lesions (one adenoma and one mucosal carcinoma) demonstrated low-frequency MSI (MSI-L). The remaining 56 neoplasms were MSI-stable (MSI-S). Six of six MSI-H, one of two MSI-L, and none of thirty MSI-S lesions showed hMLH1 hypermethylation (P

The insulin-like growth factor II receptor (IGFIIR) has been implicated as a tumor suppressor gene in human malignancy. Frequent mutation, loss of heterozygosity, and microsatellite instability (MSI) directly affecting the IGFIIR gene have been reported in several primary human tumor types. However, to our knowledge, dynamic functional evidence of a growth-suppressive role for IGFIIR has not yet been provided. We identified one MSI-positive colorectal carcinoma cell line, SW48, with monoallelic mutation in IGFIIR identical to that seen in primary colorectal carcinomas. A zinc-inducible construct containing the wild-type IGFIIR cDNA was stably transfected into SW48 cells. Growth rate and apoptosis were compared between zinc-treated, untreated, and untransfected cells. A twofold increase in IGFIIR protein expression was detected after zinc treatment in discrete clonal isolates of transfected SW48 cells. Moreover, zinc induction of exogenous wild-type IGFIIR expression reproducibly decreased growth rate and increased apoptosis. These data prove that wild-type IGFIIR functions as a growth suppressor gene in colorectal cancer cells and provide dynamic in vitro functional support for the hypothesis that IGFIIR is a human growth suppressor gene.

FHIT (fragile histidine triad gene), a candidate tumor suppressor gene, was recently identified and cloned at chromosome 3p14.2. Alterations of this gene have been reported in a number of primary human tumors, including colorectal, esophageal, gastric and lung carcinomas. However, some reports have found no abnormalities in this gene. We investigated a total of 63 primary esophageal tumors, nine esophageal cancer cell lines and 17 ulcerative colitis-associated neoplasms (UCANs) for alterations of FHIT. In 13 esophageal tumors, we employed overlapping reverse transcriptase-PCRs (RT-PCRs) to amplify and sequence the complete open reading frame of FHIT. One of 13 primary esophageal tumors analysed by RT-PCR expressed no detectable FHIT transcript; the remaining 12 expressed normal-sized transcripts with wild-type open reading frame sequences. In an additional 50 esophageal tumors, the polymorphic microsatellite loci D3S1300 and D3S1313 were used to evaluate loss of heterozygosity (LOH) at 3p14.2. Eleven of these 50 tumors showed LOH at one or both loci. In all these 11 tumors, genomic PCR and direct sequencing of FHIT exons 5-9 was performed. This analysis revealed that none of these 11 primary esophageal tumors contained any alterations in the FHIT open reading frame or adjacent intron sequences. Finally, among 17 UCANs, the in vitro synthesized protein (IVSP) assay detected no truncated protein products, nor were there any abnormalities in size or DNA sequence of FHIT RT-PCR products. However, in six of nine esophageal carcinoma cell lines, no FHIT RT-PCR product was detectable using either of the overlapping primer sets. Genomic PCR and direct sequencing of exons 5-9, also performed in these nine cell lines, revealed wild-type sequence in eight cell lines; however, one cell line contained no exon 5 PCR product. This cell line also lacked detectable FHIT transcript. These data suggest that the open reading frame of FHIT is not important in the development or progression of most primary esophageal carcinomas or UCANs, although lack of expression of the FHIT transcript may be common in esophageal cancer-derived cell lines. The possibility of an additional tumor suppressor gene at chromosome 3p14.2 remains to be evaluated.

Genomic instability at simple repeated sequences has been observed in various types of human cancers and is considered an important mechanism in tumorigenesis. Alterations at microsatellite loci have been reported scattered throughout the genome. Recently, the transforming growth factor-beta receptor type II (TGF-beta RII) and the insulin-like growth factor II receptor (IGF-IIR) genes were shown to have inactivating mutations within coding microsatellite sequences. The demonstration of mutations in two growth regulatory genes supports the idea that other regulatory genes with repeat sequences may also be targets in tumours with defective mismatch repair. We examined genes involved in tumour suppression, cell adhesion and cell cycle regulation for mutations at small repeat sequences in replication error positive gastrointestinal cancers. Several polymorphisms were found which exhibited instability, but no other instability was present in the regions examined.

The aim of this study was to detect the abnormality of the brain functional connectivity of the hypothalamus during acute spontaneous cluster headache (CH) attacks (‘in attack’) and headache-free intervals (‘out of attack’) using resting-state functional magnetic resonance imaging (RS-fMRI) technique. The RS-fMRI data from twelve male CH patients during ‘in attack’ and ‘out of attack’ periods and twelve age- and sex-matched normal controls were analyzed by the region-of-interest -based functional connectivity method using SPM5 software. Abnormal brain functional connectivity of the hypothalamus is present in CH, which is located mainly in the pain system during the spontaneous CH attacks. It extends beyond the pain system during CH attack intervals.

In this study, an inspection system for indium tin oxide (ITO) circuits has been developed. In the developed system, a polymer dispersed liquid crystal (PDLC)/ITO film is used as a sensing device to locate faulty shut/open circuits. The examined object and the PDLC/ITO film are both linked to an external power source to form an electric field. With the power on, the crystals line up, re-orientate themselves and the film covering the conducting area turns clear while the liquid crystals covering the non-conducting area are randomly scattered and diffuse light in all directions. The voltage range of the power source required to change the state of the PDLC film was estimated theoretically. Simulations were conducted to study the effects of the external power on the performance of the developed system. The results were then verified experimentally. It was shown that the developed system is a feasible system for ITO circuit inspection.

When rat (female Wistar) lungs were lavaged (bronchoalveolar lavage, BAL) six times with physiological saline, approximately the same number of alveolar macrophages (AM) were found in the first and second BAL, whereas in the third fourth, fifth, and sixth BAL, the number of AM decreased exponentially. Morphometric counting of the number of AM in histological sections of lung tissue showed that only 14% of the AM population had been recovered by BAL. Although additives to the BAL fluid such as lidocaine and/or fetal calf serum increased the AM count in the first washing considerably, the total number of AM washed out remained unaltered. Addition of the phagocytosis stimulant zymosan increased the AM count in BAL by a factor of more than 2. On stimulation of the lungs with an inert dust (silicon carbide), the AM count in the BAL and the lung was only slightly increased 8 weeks after intratracheal instillation. In contrast, after exposure to fibrogenic and cytotoxic quartz, the AM count in BAL and lung was significantly increased, and the recovery of AM had also increased by a factor of approximately 2. The experiments show that it is the micromilieu of the alveoli and the condition of the AM (certain physiological activation states, such as phagocytic activity) that essentially determine the degree of recovery.

An efficient method for the reliability analysis of systems with non-linear limit states is presented in this paper. A trust-region-based adaptive response surface is used to search for the most probable point on the limit state. The adaptive response surface construction is aided by an optimal symmetric Latin hypercube sampling-based experimental design. First-order and second-order reliability estimates using the adaptive response surface are further improved with multi-modal adaptive importance sampling. Numerical examples including a vehicle side impact problem show that the proposed method has desirable efficiency and accuracy.