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Neurofibromin 1 (NF1) is a tumor suppressor that has been previously reported to regulate RAS-MAPK signaling. The present study investigated the possible relationship between NF1 expression and anti-EGFR antibody (cetuximab) sensitivity in colorectal cancer cell lines. In addition, primary or metastatic colorectal cancer samples from patients treated with cetuximab were assessed for the association of cetuximab sensitivity. The quantities of the NF1 transcript, NF1-related pathway enrichment and NF1 mutation profile were measured and investigated using RNA sequencing and targeted DNA sequencing. Based on growth inhibition and colony formation assay results, cell lines were designated to be cetuximab-sensitive (NCI-H508 and Caco2) or cetuximab-resistant (KM12C and SM480). Western blotting revealed NF1 was highly expressed in cetuximab-sensitive cell lines whilst there was little expression in their cetuximab-resistant counterparts. Knocking down NF1 expression using small interfering RNA in the cetuximab-sensitive cell lines enhanced the phosphorylation of MEK and ERK according to western blotting. NF1 knockdown also reduced apoptosis, as observed by the decreased number of apoptotic bodies by DAPI nuclear staining and reduced cleavage of caspase and poly-(ADP ribose) polymerase. NF1 overexpression by transfection with GTPase-activating protein-related domain subunit rendered the cetuximab-resistant cell lines, KM12C and SW480, more susceptible to cetuximab-induced apoptosis. RNA sequencing of 111 RAS and BRAFV600 wild-type tumor samples collected from cetuximab-treated patients with metastatic colorectal cancer revealed that the pre-treatment NF1 expression levels were not associated with the cetuximab response. However, tumor samples obtained after cetuximab treatment displayed slightly lower NF1 transcript levels compared with those in the pre-treatment samples, suggesting that exposure to the anti-EGFR antibody may be associated with reduced NF1 expression levels. Next-generation sequencing revealed that the frequency of inactivating mutations in NF1 were rare (1.8%) in patients with colorectal cancer and were not associated with the protein expression levels of NF1 except for in a small number of cases (0.5%), where the biallelic inactivation of NF1 was observed. To conclude, the present study showed that modification of NF1 expression can affect sensitivity to cetuximab in colorectal cancer cell lines, though a limitation exists in terms of its potential application as a biomarker for RAS and BRAF V600 wild-type tumors.

Simple Summary Pancreatic cancer is a highly lethal tumor with poor prognosis. In general, pancreatic cancer is not detected in its early stages since there are no signs or symptoms. A surgical resection gives the best chance for a cure, but these lesions are detected at the terminal or metastatic stages in most patients and surgery is therefore no longer feasible. New therapeutic options are thus necessary for the pancreatic cancer treatment. Alterations to metabolic pathways have recently attracted great interest as possible cancer treatments and many studies have reported that targeting glutaminase is an ideal approach in many cancers. Here, we provide reliable evidence that pancreatic cancer requires TFEB for maintaining glutaminase-mediated glutamine metabolism, and that this is an attractive new target for pancreatic cancer therapy. Abstract Transcription factor EB (TFEB) is a master regulator of lysosomal function and autophagy. In addition, TFEB has various physiological roles such as nutrient sensing, cellular stress responses, and immune responses. However, the precise roles of TFEB in pancreatic cancer growth remain unclear. Here, we show that pancreatic cancer cells exhibit a significantly elevated TFEB expression compared with normal tissue samples and that the genetic inhibition of TFEB results in a significant inhibition in both glutamine and mitochondrial metabolism, which in turn suppresses the PDAC growth both in vitro and in vivo. High basal levels of autophagy are critical for pancreatic cancer growth. The TFEB knockdown had no significant effect on the autophagic flux under normal conditions but interestingly caused a profound reduction in glutaminase (GLS) transcription, leading to an inhibition of glutamine metabolism. We observed that the direct binding of TFEB to the GLS and TFEB gene promotors regulates the transcription of GLS. We also found that the glutamate supplementation leads to a significant recovery of the PDAC growth that had been reduced by a TFEB knockdown. Taken together, our current data demonstrate that TFEB supports the PDAC cell growth by regulating glutaminase-mediated glutamine metabolism.

Matrine is a natural compound extracted from the herb Sophora flavescens Ait which is widely used in traditional Chinese medicine for treating various diseases. Recently, matrine was reported to have antitumor effects against a variety of cancers without any obvious side effects; however, the molecular mechanisms of its antiproliferative effects on cancer are unclear. Here, we report that matrine inhibits autophagy-mediated energy metabolism, which is necessary for pancreatic cancer growth. We found that matrine significantly reduces pancreatic cancer growth in vitro and in vivo by insufficiently maintaining mitochondrial metabolic function and energy level. We also found that either pyruvate or α-ketoglutarate supplementation markedly rescues pancreatic cancer cell growth following matrine treatment. Inhibition of mitochondrial energy production results from matrine-mediated autophagy inhibition by impairing the function of lysosomal protease. Matrine-mediated autophagy inhibition requires stat3 downregulation. Furthermore, we found that the antitumor effect of matrine on pancreatic cancer growth depends on the mutation of the KRAS oncogene. Together, our data suggest that matrine can suppress the growth of KRAS-mutant pancreatic cancer by inhibiting autophagy-mediated energy metabolism.

Many cancer types are driven by oncogenic transcription factors that have been difficult to drug. Transcriptional inhibitors, however, may offer inroads into targeting these cancers. Through chemical genomics screening, we identified that Ewing sarcoma is a disease with preferential sensitivity to THZ1, a covalent small-molecule CDK7/12/13 inhibitor. The selective CDK12/13 inhibitor, THZ531, impairs DNA damage repair in an EWS/FLI-dependent manner, supporting a synthetic lethal relationship between response to THZ1/THZ531 and EWS/FLI expression. The combination of these molecules with PARP inhibitors showed striking synergy in cell viability and DNA damage assays in vitro and in multiple models of Ewing sarcoma, including a PDX, in vivo without hematopoietic toxicity.

To effectively manage research information in the field of national defense, metadata about the information should be managed systematically, and an integrated system to help convergence and management of the information should be implemented based on the metadata. In addition, the system should provide the users with effective integrated search services in a mobile environment, because searching via the use of mobile devices is increasing. The objective of this paper is to suggest a MSISS (Mobile Semantic Integrated Search System), which satisfies the requirements for effective management of the national defense research information. Specifically, we defined national defense research ontologies and national defense research rules after analyzing the Dublin Core metadata and database information of the major military research institutions. We implemented a prototype system for MSISS to demonstrate the use of the ontologies and rules for semantic integrated searching of the military research information. We also presented a triple-based search service to support semantic integrated search in a mobile environment and suggested future mobile semantic integrated search services.

To improve the diagnosis and prescription for military personnel, it is required to adopt Clinical Decision Support System (CDSS) in armed forces hospitals. The objective of this paper is to suggest a CDSS for armed forces hospitals using semantic web technologies. To this end, we designed military medical ontologies and military medical rules which consist of the various concepts and rules for supporting medical activities. We developed a semantic web-based CDSS to demonstrate the use of the ontologies and rules for treating military patients. We also showed the process of semantic search for the medical records which are created from the semantic web-based CDSS.

Being an element of plastic arts and a language of plastic forms, the color white has significant potential for improving the quality of physical environment. Colors are dealt with scientifically in chromatics, and also often become an interesting topic when analyzing a society and its history and culture. Thus, an in-depth study of the symbolism of color white and, of white pigment are required because the color has not only an important chromatic value and this should be understood a great deal, but also been the important symbol of a culture in a particular period. This paper examined the significance of the color white identified in Asian and Western cultures based on scientific and psychological understanding of the color. This also analyzed a few major white pigments, then demonstrated different methods of using the pigments and various techniques of expression used by Asian and Western painters. In addition, the paper attempted to raise awareness of the color white as chromatic color over non-color by examining the contrasting viewpoints on the color, and the changes in the viewpoints over time and place.

A series of calix[4]arene-bis-crownethers were synthesized in a fixed 1,3-alternateconformation with good yields by the reaction of amonocyclic calixcrown ether with multi-ethyleneglycoldi-p-toluenesulfonate in the presence of cesiumcarbonate. In the preparation of the monocycliccalixcrown ethers (1 and 2), the use ofpotassium carbonate as a base provided the best yieldregardless of the template concept. In two phaseextraction and competitive transport experiments forligand-metal complexation, calix[4]arene biscrown(5) provided the best selectivity for potassiumion. When a calixbiscrown ether (4) bearingdifferent sized crown ether loops coordinates to K+and Cs+, respectively, the changes of peak splittingpatterns and chemical shift on 1H NMR spectra aredependent on the complexed metal ion species.

A series of 1,3-dialkyloxycalix[4]arene dibenzocrown ethers were synthesized in the fixed 1,3-alternate conformation with good yields by the reaction of 1,3-dialkyloxycalix[4]arenes with dibenzodimesylate in the presence of caesium carbonate. The three dimensional 1,3-alternate conformation was confirmed by X-ray crystal structure. Upon two phase extraction, NMR studies on the ligand–metal complex, single ion and competitive ion transport experiments through a bulk liquid membrane, 1,3-dipropyloxycalix[4]arene dibenzocrown-6, in which two benzo groups are symmetrically attached to the crown-6 linkage, gave one of the most efficient and selective extractabilities for caesium ion over other alkali metal ions. From supported liquid membrane experiments using 1 and 3 as organic carriers, permeation coefficients of the caesium ion were estimated to be 0.42 and 0.27 cm h–1, respectively. The selectivity of caesium over sodium ion was observed to increase with permeation time.

5-Amino-2,3-dihydro-1,2,4-oxadiazol-3-one 2 was synthesized as an analogue of cytosine. Among the possible four tautomers it was established by spectroscopic methods and ab initio calculations that it exists as a lactam form 2b like cytosine. The NH is more acidic than NH2, thus it could be regioselectively mesylated and acetylated under basic conditions.

5-Amino-2-acyl-1,2,4-thiadiazolin-3-ones 2–1 can be synthesized from 5-amino-2H-1,2,4-thiadiazolin-3-one (1–1) via a selective acylation with an acid anhydride in pyridine. The 1H nmr spectral characteristics of 5-amino-2-acyl-1,2,4-thiadiazolin-3-ones 2–1 is in particular, compared with 5-amino-2H-1,2,4-thiadiazolin-3-one (1–1) and 5-amino-2-alkyl-1,2,4-thiadiazolin-3-ones 1–2, 1–3. The 5-amino group of 2–1 appeared as two peaks in its 1H nmr spectrum, which merged to a single peak at a higher temperature, while those of compound 1–1, 1–2 and 1–3 appear only as a single peak. The restricted rotation of the C(5)-N(5) (at amino) bond of 5-amino-2-acetyl-1,2,4-thiadiazolin-3-one (2a-1) is about 14.5 Kcal/mol.

Fragment pathways of 3-alkyl and acyl-5-amino-1,3,4-thiadiazolin-2-ones were completely assigned by mass analyzed kinetic energy spectra (MIKE), collisional activated dissociation (CAD) spectra and high resolution mass spectra. [NH2CS]+ and [S[dbnd]C[dbnd]O]+ ions were directly formed from molecular ion of the 3-alkyl compounds and from 5-amino-3H−1,3,4 -thiadiazolin-2-one ion ( 1 ) which was produced by McLafferty rearrangement from molecular ion of 3-acyl compounds. A loss of neutral SCO was only detected from 3-alkyl substituted molecular ions.

1,3-diproyloxycalix[4]arene azacrown ether was successfully synthesized in the fixed 1,3-alternate conformation which was confirmed by a solid state structure.

1,3-Alternate calix[4]arene crown-5 (1) and corresponding biscrown-5 (2) were synthesized and the complexation behavior with alkali metal ions examined. For both 1 and 2, potassium ion was selectively extracted from aqueous phase into organic phase over other alkali metal ions based on two phase extraction experiment. The complexation ratio between calix[4]arene biscrown-5 (2), in which two crown cavities connect to the calix[4]arene framework by 1,3-alternate fashion and potassium metal ion is found to 1:1 by proton NMR spectroscopy and extraction equilibria. Association constants (log K a ) for 1 and 2 were determined to give 2.51 and 3.49, respectively.

5-Amino-2H-1,2,4-thiadiazoline-3-one has been prepared from 2-thiobiuret through a oxidative cyclization under basic hydrogen peroxide condition. Lactam-lactim tautomerism of 5-amino-2H-1,2,4-thiadiazoline-3-one has been examined by spectroscopic methods and ab initio molecular orbital calculation. These results were shown that 5-amino-2H-1,2,4-thiadiazoline-3-one exists as lactam form. Reaction conditions for alkylating 5-amino-2H-1,2,4-thiadiazoline-3-one with alkyl halides in the presence of various bases were studied in DMF and -THF two phase system. base in DMF method gave best result in this investigation. The alkylation has been taken place at N-2 position. The identification of the product is confirmed by spectral data of IR, NMR and NMR and the comparison of authentic sample, 5-amino-2-methyl-1,2,4-thiadiazolidine-3-one, obtained from the oxidative cyclization of 5-methyl-2-thiobiuret.

1,3-diproyloxycalix[4]arene azacrown ether was successfully synthesized in the fixed 1,3-alternate conformation which was confirmed by a solid state structure.

5-Amino-2-acyl-1,2,4-thiadiazolin-3-ones 2–1 can be synthesized from 5-amino-2H-1,2,4-thiadiazolin-3-one (1–1) via a selective acylation with an acid anhydride in pyridine. The 1H nmr spectral characteristics of 5-amino-2-acyl-1,2,4-thiadiazolin-3-ones 2–1 is in particular, compared with 5-amino-2H-1,2,4-thiadiazolin-3-one (1–1) and 5-amino-2-alkyl-1,2,4-thiadiazolin-3-ones 1–2, 1–3. The 5-amino group of 2–1 appeared as two peaks in its 1H nmr spectrum, which merged to a single peak at a higher temperature, while those of compound 1–1, 1–2 and 1–3 appear only as a single peak. The restricted rotation of the C(5)-N(5) (at amino) bond of 5-amino-2-acetyl-1,2,4-thiadiazolin-3-one (2a-1) is about 14.5 Kcal/mol.

This study was conducted to investigate the effects of a no saline, a 2 ml and a 5 ml saline instillation prior to endotracheal suctioning on oxygen saturation in patients with pneumonia. The subjects in this study were 16 pneumonic patients with a tracheotomy tube, who had been admitted to the neuro-surgical intensive care unit at a university hospital in Seoul Korea. All three (0, 2 and 5 ml) saline instillation methods were applied to the 16 patients. The methods were randomly assigned to each patient. Each of the instillation methods was applied in a four-step sequence: 1) recording the level of oxygen saturation (baseline levels), 2) instilling normal saline, 3) supplying oxygen and suctioning, and 4) recording the level of oxygen saturation. The oxygen saturation was evaluated using pulse oximetry. The recovery times for oxygen saturation to return to baseline levels following suctioning were, just after suctioning, 45 seconds after suctioning and in excess of 5 minutes with 0, 2 and 5 ml saline instillations, respectively. Instillation of normal saline before suctioning could have an adverse effect on oxygen saturation, and should be used carefully as a routine intervention in patients who have pneumonia.

Background and Objectives：This study was designed to examine the effects of diamide and thioredoxin (TRX on vascular endothelial cells in order to clarify the mechanism by which vascular damage is mediated by oxygen free radicals. Materials and Methods：The pulmonary artery endothelial cell (PAEC line derived from bovine serum was cultured for 8 hours in media supplemented with various concentrations of diamide and TRX. The XTT assay, MTS assay, SRB assay, LDH activity and lipid peroxidation tests were perfomed. Results：In XTT and MTS assays, diamide significantly decreased the cell viability of cultured PAEC in a dose- and time-dependent manner. Diamide showed a decrease in the amount of total protein, although it showed an increase of lipid peroxidation and LDH activity in cultured PAEC. In regards to the protective effect of TRX on diamide-induced cytotoxicity, this showed an increase of total protein, however it showed a decrease of lipid peroxidation and LDH activity. Conclusion：Our results suggest that diamide has a vasculotoxic effect on cultured bovine PAEC and that TRX is very effective in the protection of diamide-induced cytotoxicity by duye to the increase of total protein and the decrease of lipid peroxidation and LDH activity in these cultures. (Korean Circulation J 2001;31(11 :1185-1193

Recent increase in the incidence of lung cancer often makes it difficult to differentiate between lung cancer and tuberculosis (TB), due to their radiologic similarities. Fine needle aspiration biopsy (FNAB) has been widely employed for the diagnosis of lung cancer and TB, but the diagnostic accuracy of TB is not high enough. As a rapid screening test for tuberculosis, we evaluated serological tests using Mycobacterium tuberculosis PPD and lipoarabinomannan (LAM) antigens. A total of 95 patients with indication of FNAB cytology from initial CT findings were enrolled. 25 patients had TB, 76 thoracic malignancy, and six (7.9%) of the lung cancer patients also had TB, indicating much higher prevalence of TB in thoracic tumor patients. Antibodies to PPD were elevated in 18 (72.0%) of 25 TB patients and in 22 (31.4%) of 70 patients with thoracic malignancy. In contrast, only 3 (4.7%) of 64 healthy controls aged 40 or above were seropositive to PPD antigen. The prevalence of anti-PPD antibodies in thoracic tumor patients was therefore significantly greater than that amongst the healthy controls (p