In order to detect phosphoproteins in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), an easy and fast fluorescent detection method is described. 8-Quinolinol can form ternary complexes in the gel matrix contributed by the affinity of aluminum ion to the phosphate groups on the proteins and the metal chelating property of 8-Quinolinol, exhibiting strong fluorescence in ultraviolet light. It can visualize as little as 4-8 ng of α-casein and β-casein, 15-31 ng of ovalbumin and κ-casein within 70 min. The approach utilizing 8-quinolinol could be an alternative staining method for phosphoproteomics.

A fast and matrix-assisted laser desorption/ionization-mass spectrometry compatible protein staining method in one- and two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis is described. It is based on the counterion dye staining method that employs oppositely charged two dyes, zincon and ethyl violet to form an ion-pair complex. The protocol including fixing, staining, and quick washing steps can be completed in 1-1.5 h depending upon gel thickness. It has the sensitivity comparable to the colloidal Coomassie Brilliant Blue G stain using phosphoric acid as a component of staining solution (4-8 ng). The counterion dye stain does not induce protein modifications that complicate interpretation of peptide mapping data from mass spectrometry. Considering the speed, sensitivity, and compatibility with mass spectrometry, the counterion dye stain may be more practical than any other dye-based protein stains for routine proteomic researches.

A novel fluorescent staining protocol to detect phosphoproteins in sodium dodecyl sulfate-polyacrylamide gels using a fluorescence sensor, 1-(2-hydroxy-1-naphthylazo)-2-naphthol-4-sulfonic acid sodium salt (Calcon), was developed. This method yields results within 135 min, with the sensitivities of 15 ng of α-casein and β-casein, and 62.5 ng of κ-casein, respectively. Since non-phosphoproteins have shown negative signals that are distinctly different from positive signals of phosphoproteins, this detection method allows one to monitor phosphoproteins with high specificity. Furthermore, a total protein profile can be achieved before a destaining step using a scanner with rapid and low-cost without further total protein staining.

Protein phosphorylation, one of the most important post‐translational modifications, plays critical roles in many biological processes. Thus, it is necessary to precisely detect, identify and understand the phosphoproteins from protein mixture for the study of cell biology. We introduce a sensitive and specific detection method for phosphoproteins in sodium dodecyl sulfate‐polyacrylamide gel electrophoresis (SDS‐PAGE). Anthracene Chrome Red A (ACRA) combined with the trivalent metal ion (Al 3+ ) is converted to fluorescent complex and the fluorescence is sharply increased by a change of pH environment. Phosphoproteins and non‐phosphoproteins can be easily distinguished by the fluorescence quenching due to the structural change of ACRA‐Al 3+ ‐phosphoprotein complex, unlike non‐phosphoprotein complex. The method using ACRA is a negative staining based on the fluorescence quenching and has a high sensitivity comparable to Pro‐Q Diamond stain. ACRA stain can detect 1–2 ng of α‐casein and β‐casein, 8–16 ng of ovalbumin (OVA) and κ‐casein within 130 min. Moreover, the ACRA stain showed similar linear dynamic ranges and RSD to Pro‐Q stain. The linear dynamic ranges of ACRA and the values of correlation coefficient were for OVA (8–500 ng, correlation coefficient r = 0.999), α‐casein (4–500 ng, r = 0.992), β‐casein (4–500 ng, r = 0.996), and κ‐casein (8–500 ng, 0.998), respectively. On the other hand, the values of the relative standard deviations (RSD) ranged from 2.33 to 3.56% for ACRA. The method is sensitive, specific, simple, rapid and compatible with total protein stain such as SYPRO Ruby stain. Therefore, ACRA stain can be an advanced method for phosphoprotein detection in gels.

In order to achieve an easy, rapid and sensitive protocol to detect proteins in polyacrylamide gel, an advanced negative detection method comparable to silver stain is described. When a gel was incubated with Phloxine B and followed by the development in acidic solution, the zones where forming protein-dye complex were selectively transparent, unlike opaque gel background. Within 50 min after electrophoresis, down to 0.1-0.4 ng of gel-separated proteins (similar with silver stain) could be observed, without labor-intensive and time-consuming procedure. Comparing with the most common negative stain method, Imidazole-zinc stain, Phloxine B stain has been shown higher sensitivity and distinct contrast between the transparent protein bands/spots and opaque background than those; furthermore, it is no longer necessary to concern about retention time of observation. This technique may provide a sensitive and practical choice for proteomics researches.

In this paper, we propose a cascade classifier for high-performance on-road vehicle detection. The proposed system deliberately selects constituent weak classifiers that are expected to show good performance in real detection environments. The weak classifiers selected at a cascade stage using AdaBoost are assessed for their effectiveness in vehicle detection. By applying the selected weak classifiers with their own confidence levels to another set of image samples, the system observes the resultant weights of those samples to assess the biasing of the selected weak classifiers. Once they are estimated as biased toward either positive or negative samples, the weak classifiers are discarded, and the selection process is restarted after adjusting the weights of the training samples. Experimental results show that a cascade classifier using weak classifiers selected by the proposed method has a higher detection performance.

본 논문은 네트워크 침입 탐지 시스템에서 CAM 및 해시 구조 기반 알고리듬의 비용 한계를 극복하기 위해 RAM을 이용한다. RAM을 이용한 기존 알고리듬의 다중 엔트리 처리 시 실시간 처리속도 지연 문제를 보완한 새로운 패턴 매칭기를 제안한다. 제안된 패턴 매칭기는 Merge FSM 알고리듬을 적용하여 스테이트의 수를 줄이고, RAM을 사용하기 위해 스테이트 블록과 엔트리 블록을 포함한다. 입력된 문자열과 비교할 엔트리문자열이 여러개 존재할 때 엔트리 블록에서 입력된 문자열과 엔트리 문자열들을 동시에 비교한다. 제안된 패턴 매칭기는 Snort 2.9 규칙을 이용하여 검증하였다. 실험결과 기존 탐색 방법과 비교하여 메모리 접근 빈도가 15.8% 감소하였고, 전체 메모리 크기는 2.6% 증가하였으며, 처리속도는 47.1% 증가하였다. This paper proposes a new pattern matching module to overcome the increased runtime of previous algorithm using RAM, which was designed to overcome cost limitation of hash-based algorithm using CAM (Content Addressable Memory). By adopting Merge FSM algorithm to reduce the number of state, the proposed module contains state block and entry block to use in RAM. In the proposed module, one input string is compared with multiple entry strings simultaneously using entry block. The effectiveness of the proposed pattern matching unit is verified by executing Snort 2.9 rule set. Experimental results show that the number of memory reads has decreased by 15.8%, throughput has increased by 47.1%, while memory usage has increased by 2.6%, when compared to previous methods.

In order to obtain an easy and rapid protocol to visualize phosphoproteins in SDS-PAGE, a fluorescent detection method named 8-Quinolinol (8-Q) stain is described. 8-Q can form ternary complexes in the gel matrix contributed by the affinity of aluminum ion (Al(3+) ) to the phosphate groups on the proteins and the metal chelating property of 8-Quinolinol, exhibiting strong fluorescence in ultraviolet light. It can visualize as little as 4∼8 ng of α-casein and β-casein, 16∼32 ng of ovalbumin and κ-casein which is more sensitive than Stains-All but less sensitive than Pro-Q Diamond. The protocol of 8-Q requires only 70 min in 0.75 mm mini-size or 1.0 mm large-size gels with five changes of solutions without destaining step; Pro-Q takes at least 250 min with 11 changes of solutions. In addition, the new method was confirmed by the study of dephosphorylation and LC-MS/MS, respectively. The approach to visualize phosphoprotein utilizing 8-Q could be an alternative to simplify the analytical operations for phosphoproteomics research.

High fat diets promote weight gain and are associated with the development of obesity and metabolic syndrome. Non alcoholic fatty liver disease (NAFLD), a hepatic manifestation of the metabolic syndrome, is becoming increasingly prevalent. Cystathionine- β -synthase (CBS) and cystathionine- γ -lyase (CSE) catalyze homocysteine (Hcy) metabolism via the transsulfuration pathway. These two enzymes are also responsible for hydrogen sulfide (H 2 S) production via desulfuration reactions. Both Hcy and H 2 S participate in many physiological and pathological processes. The liver, with its high expression of CBS and CSE, contributes significantly to the regulation of Hcy and H 2 S homeostasis. The objective of this study was to investigate the effect of high fat diet on liver CBS and CSE expression and its impact on Hcy and H 2 S metabolism. Mice (C57BL/6) fed a high fat diet (60% kcal fat) for 5 weeks had a greater body weight and developed fatty liver as compared to mice fed a control diet. The mRNA and protein levels of CBS and CSE in the liver were significantly elevated in mice fed a high fat diet. As a consequence, the metabolism of Hcy by CBS and CSE was markedly increased in the liver with a concomitant decrease of Hcy in the circulation. Increased CBS and CSE expression also caused a significant increase in H 2 S production in the liver. In conclusion, a high fat diet stimulated CBS and CSE expression in the liver leading to imbalanced Hcy and H 2 S homeostasis. The implication of our findings related to NAFLD is under investigation.

•An improved feature descriptor for efficient object detection is proposed.•The proposed feature can be used in replace of Haar-like feature.•It shows improved performance under a wide range of illumination conditions.

A fluorescent staining technique, using selective chelation with fluorophore and metal ion to the phosphate groups of phosphoproteins in SDS-PAGE is described. As a fluorescent dye and a metal ion, Fura 2 pentapotassium salt and Al(3+) were employed, respectively. The staining method, Fura 2 stain, has sensitivities of 16-32 ng of α-casein and β-casein, 62 ng of ovalbumin, phosvitin, and κ-casein using an ultraviolet transilluminator. Furthermore, Fura 2 stain is able to carry out continuative double detection of total proteins and phosphoproteins on the same gel within 3.5 h. Consequently, selective phosphoprotein and total protein detections could be obtained without other poststaining. Considering the low cost, simplicity, and speed, Fura 2 staining may provide great practicalities in routine phosphoproteomics research.

We propose a range estimation method for vision-based forward collision warning systems with a monocular camera. To solve the problem of variation of camera pitch angle due to vehicle motion and road inclination, the proposed method estimates virtual horizon from size and position of vehicles in captured image at run-time. The proposed method provides robust results even when road inclination varies continuously on hilly roads or lane markings are not seen on crowded roads. For experiments, a vision-based forward collision warning system has been implemented and the proposed method is evaluated with video clips recorded in highway and urban traffic environments. Virtual horizons estimated by the proposed method are compared with horizons manually identified, and estimated ranges are compared with measured ranges. Experimental results confirm that the proposed method provides robust results both in highway and in urban traffic environments.

High fat diets (HFD) are obesogenic and predispose to metabolic syndromes. Non‐alcoholic fatty liver disease (NAFLD), a hepatic manifestation of the metabolic syndrome, is characterized by hepatic steatosis with progression to steatohepatitis in advanced stages. Homocysteine (Hcy) levels are reportedly altered in patients with NAFLD, although the mechanisms responsible remain undefined. Since the liver is one of the principal sites regulating systemic Hcy levels, the objective of the present study was to investigate the effect of a HFD on hepatic Hcy metabolism. Mice were fed a control diet (10% kcal fat) or a HFD (60% kcal fat) for 5 weeks. Compared to control mice, mice fed a HFD gained more weight and developed fatty liver. Serum Hcy levels were significantly lower and corresponded with increased mRNA and protein expression of transsulfuration enzymes cystathionine‐β‐synthase and cystathionine‐γ‐lyase in the liver. Enhanced expression of transsulfuration enzymes also increased hydrogen sulfide biosynthesis in the liver of HFD fed mice. Preliminary results associate these responses to the regulation of hepatic oxidative stress. The significance of our findings as they related to obesity and NAFLD are under investigation

A fluorescence-based stain with 3,5,7,2′,4′-pentahydroxyflavone (morin hydrate, MH) was designed to stain phosphoproteins in one-dimensional sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS–PAGE). Al3+ was applied as a "fixed bridge," providing an efficient energy transfer channel between phosphoprotein and MH, to produce a strong fluorescent complex for the determination of phosphoprotein. As little as 62.5 ng of α-casein (7 or 8 phosphates) and β-casein (5 phosphates), 125 ng of ovalbumin (2 phosphates), and κ-casein (1 phosphate) could be visualized with a wide linear dynamic range. In comparison with conventional methods, MH stain is a time-saving method that takes just 90 min. It also has good compatibility with routine protein stainings such as Coomassie Brilliant Blue R (CBBR) and SYPRO Ruby for total protein analysis.

The liver plays a central role in regulating cholesterol homeostasis. High fat diets have been shown to induce obesity and hyperlipidemia. Despite considerable advances in our understanding of cholesterol metabolism, the regulation of liver cholesterol biosynthesis in response to high fat diet feeding has not been fully addressed. The aim of the present study was to investigate mechanisms by which a high fat diet caused activation of liver 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase) leading to increased cholesterol biosynthesis. Mice were fed a high fat diet (60% kcal fat) for 5 weeks. High fat diet feeding induced weight gain and elevated lipid levels (total cholesterol and triglyceride) in both the liver and serum. Despite cholesterol accumulation in the liver, there was a significant increase in hepatic HMG-CoA reductase mRNA and protein expression as well as enzyme activity. The DNA binding activity of sterol regulatory element binding protein (SREBP)-2 and specific protein 1 (Sp1) were also increased in the liver of mice fed a high fat diet. To validate the in vivo findings, HepG2 cells were treated with palmitic acid. Such a treatment activated SREBP-2 as well as increased the mRNA and enzyme activity of HMG-CoA reductase leading to intracellular cholesterol accumulation. Inhibition of Sp1 by siRNA transfection abolished palmitic acid-induced SREBP-2 and HMG-CoA reductase mRNA expression. These results suggest that Sp1-mediated SREBP-2 activation contributes to high fat diet induced HMG-CoA reductase activation and increased cholesterol biosynthesis. This may play a role in liver cholesterol accumulation and hypercholesterolemia.

본 논문은 객체검출(object detection)에 사용되는 분류기의 학습을 위한 빠르고 효율적인 Haar-like feature 선택 알고리듬을 제안한다. 기존 AdaBoost를 이용한 Haar-like feature 선택 알고리듬은 학습 샘플들에 대한 피쳐의 에러만을 고려하여 형태적으로 유사하거나 중복되는 피쳐가 선택되는 경우가 많았다. 제안하는 알고리듬은 피쳐의 형태와 피쳐간의 거리로부터 피쳐의 유사도를 계산하고 이미 선택된 피쳐와 유사도가 큰 피쳐들을 피쳐 세트에서 제거하여 빠르고 효율적인 피쳐 선택이 이루어지도록 하였다. FERET 얼굴 데이터베이스를 사용하여 제안된 알고리듬을 사용하여 학습시킨 분류기와 기존 알고리듬을 사용한 분류기의 성능을 비교하였다. 실험 결과 제안한 피쳐 선택 방법을 사용하여 학습시킨 분류기가 기존 방법을 사용한 분류기보다 향상된 성능을 보였으며, 동일한 성능을 갖도록 학습시켰을 경우 분류기의 피쳐 수가 20% 감소하였다. This paper proposes a fast and efficient Haar-like feature selection algorithm for training classifier used in object detection. Many features selected by Haar-like feature selection algorithm and existing AdaBoost algorithm are either similar in shape or overlapping due to considering only feature's error rate. The proposed algorithm calculates similarity of features by their shape and distance between features. Fast and efficient feature selection is made possible by removing selected features and features with high similarity from feature set. FERET face database is used to compare performance of classifiers trained by previous algorithm and proposed algorithm. Experimental results show improved performance comparing classifier trained by proposed method to classifier trained by previous method. When classifier is trained to show same performance, proposed method shows 20% reduction of features used in classification.

본 논문은 어플리케이션에 최적화된 ASIP설계를 하기 위해 MDL을 기반으로 한 Retargetable 컴파일러를 이용한 자동 인스트럭션 확장 시스템을 제안한다. 제안된 시스템은 어플리케이션 프로그램으로부터 얻은 정보를 이용하여 확장 가능한 인스트럭션 후보를 모두 찾는다. 확장 인스트럭션 후보는 하드웨어 라이브러리를 통해 실제 구현 시의 특성에 대한 정보를 얻게 된다. 하드웨어 특성과 수행 속도 향상을 기반으로 주어진 제한 조건에 맞게 인스트럭션 셋을 선택하고 프로세서 구조를 최적화한다. 제안된 시스템의 효용성을 확인하기 위해 다양한 벤치마크 어플리케이션을 이용하여 자동 인스트럭션 확장 시스템을 수행하였다. 제안된 시스템은 기존의 ARM9TDMI의 프로세서로부터 최적화된 인스트럭션 셋과 프로세서 구조를 갖도록 하였다. 제안된 시스템에 의해 설계된 ASIP는 주어진 제한 조건에 따라 기존 프로세서와 비교하면 평균 33.5%의 수행 사이클이 감소하는 것으로 확인되지만, 프로세서의 면적은 증가하는 것으로 측정되었다. This thesis proposes an automatic instruction extension system that utilizes retargetable compiler, based on MDL, to design an ASIP optimized for application. The proposed system uses information gathered from the application program to find all possible expandable instruction candidates. Expandable instruction candidates acquire the realization characteristics through hardware library. The system chooses instruction set and optimizes processor structure satisfying constraints on the bases of hardware characteristics and increase in execution speed. To confirm the efficiency of the proposed system, automatic instruction extension system was performed using various benchmark applications. The proposed system acquired optimized instruction set and processor structure, which are expanded from the commercial version of ARM9TDMI. Experimental results show that number of execution cycle has been reduced by 33.5% when compared to conventional version of ARM9TDMI, while area has been slightly increased.

Hypercholesterolemia is associated with an increased risk of coronary artery disease, such as atherosclerosis. The liver plays a central role in regulating cholesterol homeostasis. High fat diet has been shown to induce obesity and hyperlipidemia. Despite considerable advance in our understanding of cholesterol metabolism, the regulation of liver cholesterol biosynthesis in response to high fat diet feeding has not been fully addressed. The aim of the present study was to investigate mechanisms by which a high fat diet caused activation of hepatic 3-hydroxy-3-methylglutaryl coenzyme A reductase (HMG-CoA reductase), the rate-limiting enzyme, leading to increased cholesterol biosynthesis.

High fat diets, particularly those rich in cholesterol and saturated fatty acids promote weight gain and are associated with the development of obesity, the metabolic syndrome and cardiovascular disease (CVD). Non-alcoholic fatty liver disease (NAFLD), a hepatic manifestation of the metabolic syndrome, is becoming increasingly prevalent. NAFLD covers a broad spectrum of disease characterized by hepatic steatosis (lipid accumulation) which may progress to steatohepatits and cirrhosis in more advanced stages. Clinical studies suggest that NAFLD may increase ones risk of future CVD events, independent of classical risk factors. Elevated levels of circulating homocysteine (Hcy), an intermediate amino acid formed in methionine metabolism, has also been implicated in several vascular abnormalities. Studies have shown that plasma Hcy levels may be altered in patients with NAFLD. The mechanisms responsible await elucidation. Given that the liver is one of the primary sites regulating plasma Hcy levels, the objective of the present study was to investigate the effect of a high fat diet on Hcy metabolism in the liver.

본 논문은 4-Way 캐쉬의 선택된 element만을 사용하여 어플리케이션 수행 사이클을 줄인 향상된 동적 분기 예측기를 제안한다. 제안된 동적 분기 예측기는 분기명령어가 페치되면 MRU 버퍼를 참조하여 4-Way 캐쉬의 선택된 element에서 타깃 주소를 얻으므로, 모든 element에 접근하는 기존의 동적 분기 예측기보다 제한된 전력하에서 BTAC entry 수를 증가시킬 수 있어 분기 예측 성공률과 어플리케이션의 수행속도가 상당히 향상된다. 제안된 동적 분기 예측기의 효율성을 SMDL 시스템에 의해 생성된 코어가 벤치마크 어플리케이션을 수행하여 검증한다. 실험결과 동적 분기 예측기가 없는 코어에 비해 생성된 코어의 어플리케이션 수행 사이클은 평균 10.1% 감소하고 어플리케이션의 전력소모는 7.4% 증가한다. 기존 동적 분기 예측기를 사용하는 코어에 비해 수행 사이클은 평균 4.1% 줄어든다. This paper proposes an improved branch predictor that reduces the number execution cycles of applications by selectively accessing a specific element in 4-way associative cache. When a branch instruction is fetched, the proposed branch predictor acquires a branch target address from the selected element in the cache by referring to MRU buffer. Branch prediction rate and application execution speed are considerably improved by increasing the number of BTAC entries in restricted power condition, when compared with that of previous branch predictor which accesses all elements. The effectiveness of the proposed dynamic branch predictor is verified by executing benchmark applications on the core simulator. Experimental results show that number of execution cycles decreases by an average of 10.1%, while power consumption increases an average of 7.4%, when compared to that of a core without a dynamic branch predictor. Execution cycles are reduced by 4.1% in comparison with a core which employs previous dynamic branch predictor.