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Thermo-mechanical analysis of monolithic microwave integrated circuit (MMIC) packaging is essential to guarantee the reliability of radio frequency/microwave applications. However, a method for fast and accurate analysis of MMIC packaging structures has not been developed. Here, we demonstrate a machine learning (ML)-based solution for thermo-mechanical analysis of MMIC packaging. This ML-based solution analyzes temperature and thermal stresses considering 13 design parameters categorized into material properties, geometric characteristics, and thermal boundary conditions. Finite element simulation with the Monte Carlo method is utilized to prepare 40,000 data samples for supervised learning and validation of the ML solution, and a laser-assisted thermal experiment verifies the accuracy of the simulation. After data preparation, regression tree ensemble and artificial neural network (ANN) learning models are investigated. The results indicate that the ANN models accurately predict the temperature and thermal stresses, showing a 1.69 % minimum error. Finally, the developed ML solution is deployed as a web application format for facile approaches. We believe that this study will provide a guideline for developing ML-based solutions in chip packaging design technology.

In the real world, the two challenges of stereo vision system include a robust system under various radiometric changes and real-time process. To extract depth information from stereoscopic images, this paper proposes Patchmatch-based robust and fast stereo matching under radiometric changes. For this, a cost function was designed and minimized for estimating an accurate disparity map. Specifically, we used a prior probability to minimize the occlusion region and a smoothness term that considers convexity of objects to extract a fine disparity map. For evaluating the performance of the proposed scheme, we used Middlebury stereo data sets with radiometric changes. The experimental result showed that the proposed method outperforms state-of-the-art methods by up to 3.35 percent better and a range of 4.71 - 27.24 times faster result in terms of bad pixel error and processing time, respectively. Therefore, we believe that the proposed scheme can be a useful tool for computer vision-based applications.

Natural killer cells (NK cells) are cells that destroy foreign and malignant cells from our bodies. The distribution of immune cells (i.e., the ratio of T, B and NK cells) and the number as well as activity of NK cells vary from person to person. These differences provide the basis for the diversity of responsiveness to disease that is observed in a population. In addition, the immune system is influenced by the environment. Notably, the number of immune cells decreases with stress, and the number and activity of immune cells decreases when they get older. The balance of NK cell activity in the body seems to be the most important for being healthy. NK cells belong to a group of large granular lymphocytes; they play an important role in the immune system against malignant and infected targets. Unlike T cells, NK cells act as innate immune cells, showing rapid and selective ‘natural' killing of cellular targets (tumour cells and infected cells) with neither any need for major histocompatibility complex class I expression nor any sensitization step. Despite the innate property of NK cells to kill infected or malignant cells when rapid immune response is required being a great advantage, the ready-to-kill status of NK cells could have detrimental effects as well. Therefore, NK cell activation needs to be tightly modulated by the activation of inhibitory receptors, and this signalling balance orders NK cells whether to kill target cells or to stay in an inactive state. Despite sufficient evidence that NK cells fight tumour cells, pharmaceutical treatment approaches that target NK cells remain few in number. Nevertheless, the possibility of NK cells to be used as therapeutic agent is increasingly supported by recently approved cancer cell-targeting therapies, which proved that it is possible to appropriately regulate NK cell activity. The immune system can be considered as a health barometer in that when a disease occurs, the immune system is activated; NK cells are at the heart of this immunomodulation.

Recently, the cost-volume filtering (CVF) methods for local stereo matching have provided fast and accurate results compared to those of the other method. However, CVF still causes incorrect results in the occlusion and texture-free regions. In particular, cost aggregation by pixel units involves complex computation because of its dependence on the image resolution and search range. This paper presents a robust stereo matching method for occluded regions. First, we generate cost volumes using the CENSUS transform and the scale-invariant feature transform (SIFT). Then, label-based cost volumes are aggregated using adaptive support weight and the simple linear iterative clustering (SLIC) scheme from two generated cost volumes. In order to obtain optimal disparity by two label-based cost volumes, we select the disparity corresponding to high confidence similarity of CENSUS or SIFT with minimum cost point. Experimental results show that our method estimates the optimal disparity in occlusion information, which exists only in the scene of one of the stereo pairs.

This paper presents an effective method to reduce computational cost of Cost-volume filtering for stereo matching. We first obtain initial depth values by using pixel-wise cost volume and estimate principal depth values for reducing computational cost. The proposed method can be applied to advanced driver assistance system, object recognition, and other algorithms that require 3-D depth information.

One of the major goals of the Chromosome-Centric Human Proteome Project (C-HPP) is to fill the knowledge gaps between human genomic information and the corresponding proteomic information. These gaps are due to “missing” proteins (MPs)—predicted proteins with insufficient evidence from mass spectrometry (MS), biochemical, structural, or antibody analyses—that currently account for 2579 of the 19587 predicted human proteins (neXtProt, 2017–01). We address some of the lessons learned from the inconsistent annotations of missing proteins in databases (DB) and demonstrate a systematic proteogenomic approach designed to explore a potential new function of a known protein. To illustrate a cautious and strategic approach for characterization of novel function in vitro and in vivo, we present the case of Na(+)/H(+) exchange regulatory cofactor 1 (NHERF1/SLC9A3R1, located at chromosome 17q25.1; hereafter NHERF1), which was mistakenly labeled as an MP in one DB (Global Proteome Machine Database; GPMDB, 2011–09 release) but was well known in another public DB and in the literature. As a first step, NHERF1 was determined by MS and immunoblotting for its molecular identity. We next investigated the potential new function of NHERF1 by carrying out the quantitative MS profiling of placental trophoblasts (PXD004723) and functional study of cytotrophoblast JEG-3 cells. We found that NHERF1 was associated with trophoblast differentiation and motility. To validate this newly found cellular function of NHERF1, we used the Caenorhabditis elegans mutant of nrfl-1 (a nematode ortholog of NHERF1), which exhibits a protruding vulva (Pvl) and egg-laying-defective phenotype, and performed genetic complementation work. The nrfl-1 mutant was almost fully rescued by the transfection of the recombinant transgenic construct that contained human NHERF1. These results suggest that NHERF1 could have a previously unknown function in pregnancy and in the development of human embryos. Our study outlines a stepwise experimental platform to explore new functions of ambiguously denoted candidate proteins and scrutinizes the mandated DB search for the selection of MPs to study in the future.

Robust stereo matching under radiometric changes is a necessary method to use stereo vision in a real situation. This paper proposes a fast and robust stereo matching under various radiometric changes by using Census transform. Experimental result shows that proposed method has a better performance in terms of computational time and bad pixel error.

Surgical resection of gastric cancer has made significant progress, but majority of patients with advanced gastric cancer face relapse and die within five years. In this study, the antitumor activity of ex vivo expanded T cells against the human gastric cancer was evaluated in vitro and in vivo. Human peripheral blood mononuclear cells were cultured with IL-2-containing medium in anti-CD3 antibody-coated flasks for 5 days, followed by incubation in IL-2-containing medium for 9 days. The resulting populations were mostly CD3(+) T cells (97%) and comprised 1% CD3⁻CD56(+), 36% CD3(+)CD56(+), 11% CD4(+), and 80% CD8(+). This heterogeneous cell population was also called cytokine-induced killer (CIK) cells. CIK cells strongly produced IFN-γ, moderately TNF-α, but not IL-2 and IL-4. At an effector-target cell ratio of 30:1, CIK cells destroyed 58% of MKN74 human gastric cancer cells, as measured by the ⁵¹Cr-release assay. In addition, CIK cells at doses of 3 and 10 million cells per mouse inhibited 58% and 78% of MKN74 tumor growth in nude mouse xenograft assays, respectively. This study suggests that CIK cells may be used as an adoptive immunotherapy for gastric cancer patients.

Malignant glioma is the most common primary brain tumor in adults and the median survival for patients is less than a year. Despite aggressive treatments including surgical resection, radiotherapy, and chemotherapy, only modest improvement has been achieved in the survival of patients with glioma. In this study, the antitumor activity of cytokine-induced killer (CIK) cells against human glioma cancer was evaluated in vitro and in vivo. Human peripheral blood mononuclear cells were cultured with IL-2-containing medium in anti-CD3 antibody-coated flasks for 5 days, followed by incubation in IL-2-containing medium for 9 days. The number of cells increased more than 200-fold and the viability was90%. The resulting populations were consisted of 96% CD3(+), 2% CD3(-)CD56(+), 68% CD3(+)CD56(+), 2% CD4(+),1% CD4(+)CD56(+), 80% CD8(+), and 49% CD8(+)CD56(+). This heterogeneous cell population was called as CIK cells. At an effector-target cell ratio of 30:1, CIK cells destroyed 43% of U-87 MG human glioma cells, as measured by the (51)Cr-release assay. In addition, CIK cells at doses of 0.3, 1, and 3 million cells per mouse inhibited 23%, 40%, and 50% of U-87 MG tumor growth in nude mouse xenograft assays, respectively. This study suggests that CIK cells may be used as an adoptive immunotherapy for glioma cancer patients.

Cervical cancer is a major cause of cancer mortality in women worldwide and is an important public health problem for adult women in developing countries. Despite aggressive treatment with surgery and chemoradiation, the outcomes for cervical cancer patients remain poor. In this study, the antitumor activity of cytokine-induced killer (CIK) cells against human cervical cancer was evaluated in vitro and in vivo. Human peripheral blood mononuclear cells were cultured with IL-2-containing medium in anti-CD3 antibody-coated flasks for 5 days, followed by incubation in IL-2-containing medium for 9 days. The resulting populations of CIK cells comprised 95% CD3(+), 3% CD3(-)CD56(+), 35% CD3(+)CD56(+), 11% CD4(+)

Cytokine-induced killer (CIK) cells are ex vivo expanded T cells with natural killer cell phenotypes and functions. In this study, the anti-tumor activity of CIK cells against hepatocellular carcinoma was evaluated in vitro and in vivo. In the presence of anti-CD3 antibody and IL-2 for 14 days, human peripheral blood mononuclear cell population changed to heterogeneous CIK cell population, which comprised 96% CD3(+), 3% CD3( inverted exclamation mark(c))CD56(+), 32% CD3(+)CD56(+), 11% CD4(+), 75% CD8(+), and 30% CD8(+)CD56(+). CIK cells produced significant amounts of IFN-gamma and TNF-alpha; however, produced only slight amounts of IL-2, IL-4, and IL-5. At an effector-target cell ratio of 30:1, CIK cells destroyed 33% of SNU-354 human hepatocellular carcinoma cells, which was determined by the (51)Cr-release assay. In addition, a dose of 1x10(6) CIK cells per mouse inhibited 60% of SNU-354 tumor growth in irradiated nude mice. This study suggests that CIK cells may be used as an adoptive immunotherapy for patients with hepatocellular carcinoma.

Activity of the STE20-related kinase hMINK was investigated. hMINK was expressed widely, though not ubiquitously, in human tissues: highest levels being found in haematopoietic tissues but also in brain, placenta, and lung. Mutagenesis revealed that T-191. and Y-193 in the substrate recognition loop of the catalytic domain were critical for kinase activity against exogenous substrates and autophosphorylation. A mutation on T-187 showed reduced enzymatic activity against exogenous substrates but retained autophosphorylationactivity. Phosphorylation was confirmed by the use of a phospho-specific T-187 antibody. hMINK activated the JNK signal transduction pathway and optimal JNK activation occurred when the C-terminus was deleted. In addition, overexpression of the C-terminal domain devoid of kinase activity also resulted in significant activation of the JNK pathway. These data suggest that hMINK requires an activation step that dissociates the C terminal, thereby freeing the catalytic domain to interact with substrates. Models for receptor-mediated activation of hMINK are discussed. (C) 2002 Elsevier Science (USA). All rights reserved.

Splicing variants of type 4 phosphodiesterases (PDE4) are regulated by phosphorylation. In these proteins, a conserved region is located between the amino-terminal domain, which is the target for phosphorylation, and the catalytic domain. Previous studies have indicated that nested deletions encompassing this region cause an increase in catalytic activity, suggesting this domain exerts an inhibitory constraint on catalysis. Here, we have further investigated the presence and function of this domain. A time-dependent increase in hydrolytic activity was observed when PDE4D3 from FRTL-5 cells was incubated with the endoproteinase Lys-C. The activation was abolished by protease inhibitors and was absent when a phosphorylated enzyme was used. Western blot analysis with PDE4D-specific antibodies indicated the Lys-C treatment separates the catalytic domain of PDE4D3 from the inhibitory domain. Incubation with antibodies recognizing an epitope within this domain caused a 3- to 4-fold increase in activity of native or recombinant PDE4D3. Again, PDE activation by these antibodies had properties similar to, and not additive with, the activation by protein kinase A phosphorylation. An interaction between the inhibitory domain and both regulatory and catalytic domains of PDE4D3 was detected by the yeast two-hybrid system. Mutations of Ser54 to Ala in the regulatory domain decreased or abolished this interaction, whereas mutations of Ser54 to the negatively charged Asp strengthened it. These data strongly support the hypothesis that an inhibitory domain is present in PDE4D and that phosphorylation of the regulatory domain causes activation of the enzyme by modulating the interaction between inhibitory and catalytic domains.

Three potential fluorogenic substrates for assay of aminopeptidase P (AP-P) have been prepared and evaluated, using enzyme purified from porcine kidney. They are based on internal quenching of the synthetic, fluorescent amino acid (R, S)-2-amino-3-(7-methoxy4-coumaryl)propanoic acid ((R,S)-Amp) by a 2, 4dinitrophenyl (DNP) group. The compounds are X-Pro-Pro-(R, S)-Amp-NH2, where X is H-Lys(epsilon-DNP), H-Orn(delta-DNP), or L-2-amino-3-(DNP)aminopropionic acid. The first two were found to be excellent substrates for AP-P, with respective Km values of 4.8 and 5.2 microM. An advantageous feature is that under the conditions of assay, using 4-mm2 cells, the substrates are without noticeable quenching effect on the fluorescence of Pro-Pro-(R,S)-Amp-NH2 (the product liberated by the action of AP-P). At concentrations greater than about 30-50 microM, both substrates appear to inhibit the enzyme, but this has little practical consequence since assays can be carried out at substrate concentrations, giving up to approximately 80% of Vmax without this inhibitory effect being noticeable. The Lys derivative was found to be a very useful substrate for a continuous assay for AP-P and equally good in a discontinuous assay of multiple samples using microtiter plates. The racemic center at the Amp residue did not prevent total hydrolysis of the Lys derivative, suggesting that subsite specificity in AP-P does not extend as far as the P3' position.

The presence of active immunity within the brain supports the possibility of effective immunotherapy for glioblastoma (GBM). To provide a clinically-relevant adoptive immunotherapy for GBM using ex vivo expanded cytokine-induced killer (CIK) cells, the treatment capability of CIK cells, either alone or in combination with temozolomide (TMZ) were evaluated. Human CIK (hCIK) cells were cultured from PBMC using activating anti-CD3 antibody and IL-2, which were 99% CD3+, 91% CD3+CD8+ and 29% CD3+CD56+. In vitro, hCIK cells showed tumor-specific cytotoxicity against U-87MG human GBM cells. When hCIK cells were injected into tail veins of immune-compromised mice bearing U-87MG tumors in their brains, numerous CIK cells infiltrated into the brain tumors. CIK treatments (1x10(5), 1x10(6) or 1x10(7), once a week for four weeks) inhibited the tumor growth significantly in a dose-dependent manner; 44, 54 and 72% tumor volume reduction, respectively, compared with the control group (P

The scheme proposed in this thesis organizes dynamically clusters at the time of events occurrence and maintains them. The other side, the existing cluster based USN (ubiquitous sensor networks) organize at the network initial time in a lump. So the existing method causes of much energy exhaustion. Therefore the scheme proposed in this thesis can be more efficient than the existing one in the aspect of the energy usage and the gathering and sending data. In order to efficiently process various events, the scheme proposed in this thesis divides a cluster when the other events occurred in the area which the cluster organized in. In another case, it merges clusters when a same event occurs in a different near cluster. Like above, the scheme of this thesis dynamically organizes clusters in the node centeredness. The simulation to evaluating the scheme shows that the scheme of this thesis increase the performance by maximumly 50% compared with the existing study.

Despite the recent improvement in the treatment of ovarian cancer, this disease is still leading cause of cancer death in women. In this study, the anti-tumor activity of cytokine-induced killer (CIK) cells against human ovarian cancer was evaluated in vitro and in vivo. Although CD3+CD56+ cells were rare in fresh human peripheral blood mononuclear cells, they could expand more than 1,000-fold on day 14 in the presence of anti-CD3 antibody plus IL-2. At an effector-target cell ratio of 30:1, CIK cells destroyed 45% of SK-OV-3 human ovarian cancer cells, which was determined by the 51Cr-release assay. In addition, CIK cells at a dose of 23 million cells per mouse inhibited 73% of SK-OV-3 tumor growth in nude mouse xenograft assay. This study suggests that CIK cells may be used as an adoptive immunotherapy for patients with ovarian cancer.

Lung cancer is the leading cause of cancer-related death among men and women in the world. Despite the aggressive treatment with surgery, radiation and chemotherapy, the long term survival for lung cancer patients remains low. In this study, the anti-tumor activity of cytokine-induced killer (CIK) cells against human lung cancer was evaluated in vitro and in vivo. Although CD3(+)CD56(+) CIK cells were rare in fresh human peripheral blood mononuclear cells, they could expand more than 1000-fold on day 14 in the presence of anti-CD3 antibody plus IL-2. At an effector-target cell ratio of 30:1, CIK cells destroyed 98% of NCI-H460 human lung cancer cells, which was determined by the (51)Cr-release assay. In addition, CIK cells at doses of 3 and 30 million cells per mouse inhibited 57% and 77% of NCI-H460 tumor growth in nude mouse xenograft assay, respectively. This study suggests that CD3(+)CD56(+) CIK cells may be used as an adoptive immunotherapy for patients with lung cancer.

The membrane-bound form of mammalian aminopeptidase P (AP-P; EC 3.4. 11.9) is a mono-zinc-containing enzyme that lacks any of the typical metal binding motifs found in other zinc metalloproteases. To identify residues involved in metal binding and catalysis, sequence and structural information was used to align the sequence of porcine membrane-bound AP-P with other members of the peptidase clan MG, including Escherichia coli AP-P and methionyl aminopeptidases. Residues predicted to be critical for activity were mutated and the resultant proteins were expressed in COS-1 cells. Immunoelectrophoretic blot analysis was used to compare the levels of expression of the mutant proteins, and their ability to hydrolyze bradykinin and Gly-Pro-hydroxyPro was assessed. Asp449, Asp460, His523, Glu554, and Glu568 are predicted to serve as metal ion ligands in the active site, and mutagenesis of these residues resulted in fully glycosylated proteins that were catalytically inactive. Mutation of His429 and His532 also resulted in catalytically inactive proteins, and these residues, by analogy with E. coli AP-P, are likely to play a role in shuttling protons during catalysis. These studies indicate that mammalian membrane-bound AP-P has an active-site configuration similar to that of other members of the peptidase clan MG, which is compatible with either a dual metal ion model or a single metal ion in the active site. The latter model is consistent, however, with the known metal stoichiometry of both the membrane-bound and cytosolic forms of AP-P and with a recently proposed model for methionyl aminopeptidase.

The plasma membrane of many cell types contains a cohort of peptidases that serve to modulate the activity of circulating regulatory peptides. Many of these are zinc metallopeptidases and these enzymes are particularly abundant in the brush border membranes of renal and intestinal microvilli. Some of them such as neprilysin (NEP; EC 3.4.24.11) and aminopeptidase N (AP-N; EC 3.4.11.2) also exist as cluster differentiation antigens (CD10 and CD 13 respectively) on the surface of leukocytes and may play a role in regulation of the immune system1. Both NEP and AP-N also have roles in the metabolism of certain cardiovascular and neuropeptides, e.g. natriuretic peptides2 and enkephalins3. NEP and AP-N are type ll integral membrane proteins and possess the typical HExxH zincin motif4 characteristic of many zinc peptidases. Recently, we have focused on another brush border zinc peptidase, aminopeptidase P (AP-P; X-Pro aminopeptidase; EC 3.4.11.9) which we showed to be unusual among the membrane peptidases in being anchored to the membrane by a glycosylphosphatidylinositol (GPI) moiety5. AP-P also shows a number of other atypical features which led us to attempt the molecular cloning of the pig kidney enzyme.

Aminopeptidase P (AP-P), purified to homogeneity from porcine kidney membranes, was completely inactivated by treatment with 0.2 mM diethylpyrocarbonate (DEP) at pH 7.0. Treatment of the modified enzyme with 20 mM hydroxylamine resulted in recovery of AP-P activity. The differential absorption of native and modified AP-P at 240 nm showed that DEP modified two histidyl residues per mol of AP-P. The substrates, bradykinin(1–5) and Gly-Pro-Hyp, and also the inhibitor, apstatin, could protect against DEP inactivation. These results suggest that histidine residues are critical for AP-P activity.

Human bone marrow mesenchymal stem cells (MSC) are multipotent progenitors that generate osteoblasts, chondrocytes, adipocytes, myoblasts and the bone marrow stromal cells that support hematopoiesis. Although the bone marrow microenvironment is hypoxic, little is known about the maintenance and response of MSC and their bone marrow stromal progeny to hypoxia. Using cDNA microarray hybridization technologies, we show, for the first time, that a total of 231 mRNAs in cultured MSCs are regulated by short exposures (4–48hrs) to hypoxia. These include known hypoxia-responsive genes, such as BHLHB2, PGK1, GLUT-1 and VEGF. Interestingly, we demonstrate that a significant proportion of genes involved in cell growth, proliferation or survival are also regulated by hypoxia in these cells. Amongst these, we have identified the centromeric protein CENP-F as a novel gene up-regulated in cultured MSCs by hypoxia. This up-regulation results in an increased level of CENP-F protein. The hypoxic stimulus also enhances cell division in the bone marrow MSC-derived stromal cells, resulting in a doubling in cell number within 24hrs. This contrasts with the effects of hypoxia on mature endothelial cells (EC), where recruitment into cell cycle is unchanged. Our demonstration that hypoxia promotes cell division and cell proliferation in the bone marrow stromal progeny of human MSC may have wider implications for the regulated growth and survival of other stem cells in hypoxic microenvironmental niches.