• Title/Summary/Keyword: GLC

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Identification of the Pig β-1,3-N-acetylglucosaminyltransferase 1 (pB3GNT1) that is Involved in Poly-N-acetyllactosamine (poly-LacNAc) Synthesis (Poly-N-acetyllactosamine (poly-LacNAc) 합성에 관여하는 돼지 β-1,3-N-acetylglucosaminyltransferase I (pB3GNT1) 유전자 동정)

  • Kim, Ji-Youn;Hwang, Hwan-Jin;Chung, Hak-Jae;Hochi, Shinichi;Park, Mi-Ryung;Byun, Sung June;Oh, Keon Bong;Yang, Hyeon;Kim, Kyung-Woon
    • Journal of Life Science
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    • v.28 no.4
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    • pp.389-397
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    • 2018
  • The structure of glycan residues attached to glycoproteins can influence the biological activity, stability, and safety of pharmaceutical proteins delivered from transgenic pig milk. The production of therapeutic glycoprotein in transgenic livestock animals is limited, as the glycosylation of mammary gland cells and the production of glycoproteins with the desired homogeneous glycoform remain a challenge. The ${\beta}$-1,3-N-acetylglucosaminylatransferase1 (B3GNT1) gene is an important enzyme that attaches N-acetylglucosamine (GlcNAc) to galactose (Gal) residues for protein glycosylation; however, there is limited information about pig glycosyltransferases. Therefore, we cloned the pig B3GNT1 (pB3GNT1) and investigated its functional properties that could attach N-acetylglucosamine to galactose residue. Using several different primers, a partial pB3GNT1 mRNA sequence containing the full open reading frame (ORF) was isolated from liver tissue. The ORF of pB3GNT1 contained 1,248 nucleotides and encoded 415 amino acid residues. Organ-dependent expression of the pB3GNT1 gene was confirmed in various organs from adult and juvenile pigs. The pB3GNT1 mRNA expression level was high in the muscles of the heart and small intestine but was lower in the lungs. For functional characterization of pB3GNT1, we established a stable expression of the pB3GNT1 gene in the porcine kidney cell line (PK-15). As a result, it was suggested that the glycosylation pattern of pB3GNT1 expression in PK-15 cells did not affect the total sialic acid level but increased the poly N-acetyllactosamine level. The results of this study can be used to produce glycoproteins with improved properties and therapeutic potential for the generation of desired glycosylation using transgenic pigs as bioreactors.

Comparison of Clinical Features of 11 Korean Patients with Mucolipidosis II and III Including a Case of Mucolipidosis II with a Novel Mutation of GNPTAB (새로운 GNPTAB 유전자 돌연변이로 진단된 뮤코지방증 2형 1례를 포함한 국내 뮤코지방증 환자의 임상적 특징에 대한 분석)

  • Kim, Jinsup;Yang, Misun;Yang, Aram;Cho, Eun Hye;Park, Hyung-Doo;Sohn, Young Bae;Cho, Sung Yoon;Jin, Dong-Kyu
    • Journal of The Korean Society of Inherited Metabolic disease
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    • v.17 no.3
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    • pp.85-91
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    • 2017
  • Purpose: The aim of this study was to describe the clinical and biochemical features as well as the molecular analysis of a newly diagnosed illustrative case with ML II and to analyze the clinical features of 11 Korean patients with ML II/III. Method: Including a newly diagnosed patient, total 11 patients in 10 families were diagnosed as ML II (n=7) or ML III (n=4) were enrolled in the study. A diagnosis of ML II or III was made by demonstrating increased lysosomal enzyme activities in the plasma and sequence analysis of GNPTAB with characteristic clinical features. Result: A illustrative case of ML II patient was a 17 month-old boy showing characteristic facial appearance, multiple joint contractures with cardiac involvements. The enzyme assay showed increased lysosomal enzyme activities in the plasma. We identified compound heterozygous mutations in GNPTAB sequence analysis, including a frameshift (c.3428dupA [pAsn1143Lysfs*3]) and a nonsense variant c.673C>T (p.Gln225*). In total 11 patients with ML II/III, the patients with ML II showed severe growth retardation (height standard deviation score -3.2 [${\pm}1.5$]), compare to patients with ML III. Furthermore, patients with ML II patients had serious cardiac problem (n=4), hepatomegaly (n=3) and underwent tracheostomy (n=3) with further respiratory support due to respiratory distress. To improve osteoporosis and bone pain, all patients with ML III and four of 7 patients with ML II treated with intravenous pamidronate. Conclusion: Here we showed a newly diagnosed case of ML II and clinical features of 11 Korean patients with ML II or III. These data could be helpful for further diagnosis of mucolipidosis, a rare inherited metabolic disease, in Korea.

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The analysis of physical features and affective words on facial types of Korean females in twenties (얼굴의 물리적 특징 분석 및 얼굴 관련 감성 어휘 분석 - 20대 한국인 여성 얼굴을 대상으로 -)

  • 박수진;한재현;정찬섭
    • Korean Journal of Cognitive Science
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    • v.13 no.3
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    • pp.1-10
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    • 2002
  • This study was performed to analyze the physical attributes of the faces and affective words on the fares. For analyzing physical attributes inside of a face, 36 facial features were selected and almost of them were the lengths or distance values. For analyzing facial contour 14 points were selected and the lengths from nose-end to them were measured. The values of these features except ratio values normalized by facial vortical length or facial horizontal length because the face size of each person is different. The principal component analysis (PCA) was performed and four major factors were extracted: 'facial contour' component, 'vortical length of eye' component, 'facial width' component, 'eyebrow region' component. We supposed the five-dimensional imaginary space of faces using factor scores of PCA, and selected representative faces evenly in this space. On the other hand, the affective words on faces were collected from magazines and through surveys. The factor analysis and multidimensional scaling method were performed and two orthogonal dimensions for the affections on faces were suggested: babyish-mature and sharp-soft.

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Decomposition of a Substituted Diphenyl Ether Herbicide(MC-4379) by Some Environmental Factors (환경요인(環境要因)에 의(依)한 치환(置換) Diphenyl Ether계(系) 제초제(除草劑) MC-4379의 분해(分解)에 관(關)한 연구(硏究))

  • Lee, Jae-Koo;Kim, Hae-Yong
    • Applied Biological Chemistry
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    • v.18 no.1
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    • pp.30-41
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    • 1975
  • A substituted diphenyl ether herbicide(MC-4379) was studied on the decomposition by some environmental factors; sunlight, microorganisms, and the crude enzyme in rice plant extract. All the decomposition products were confirmed by means of TLC, GLC, and IR. The parent compound and the decomposition products were put to the test for the effect on the growth of some plants. The results obtained are summarized as follows: 1. Photolysis Amino-MC-4379, 2,4-dichloro-3'-carboxyl-4'-nitrodiphenyl ether, Nitrofen, 2,4-dichloro-3'-carboxyl-4'-amino-diphenyl ether, amino-Nitrofen, 3-carboxymethyl-4-nitrophenol, p-nitrophenol, p-aminophenol, etc. were confirmed as photoproducts, in addition to a relatively small amount of an unknown compound. It was confirmed that the solution-phase photolysis of MC-4379 was accelerated much more by the aid of a photosensitizer benzophenone. 2. Degradation by the crude extract of germinating rice seeds Nitrofen was confirmed as a major degradation product, in addition to a relatively small amount of and unknown compound. Most of the parent compound remained unchanged. 3. Degradation by microorganisms Nitrofen and amino-MC-4379 were confirmed as the major products. in addition to a small amount of an unknown compound. 4. The germinating rice seeds and soybean were grown in the 1,000 ppm emulsions of some chemicals, respectively. The effect on rice plant growth was in the inhibitory order of p-nitrophenol > C-6989> Nitrofen > amino-Nitrofen > MC-4379. The effect on soybean was in the order of Nitrofen > amino-Nitrofen > MC-4379. Two weeds, Amantus blitum and Setaria viridis were grown in the 500 ppm emulsions containing the compounds, respectively. After incubation for 3 days, it was observed that all the shoots had been dead.

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Fermentation Characteristics of Cheese Slurry prepared from Caseinates (카세인을 이용한 치즈곤죽의 발효특성)

  • Jang, Hae-Dong;Lee, Hyong-Joo
    • Korean Journal of Food Science and Technology
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    • v.17 no.5
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    • pp.389-398
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    • 1985
  • To shorten the processing of cheese slurry, four different slurries, ie, Control, Cheddar 1 and 2, and Italian-type that were made of Na-caseinates, cream, trace elements, lactic culture, and enzymes were fermented at $30^{\circ}C$ for 7days with daily stirring. PH, titratable acidity, soluble nitrogen, viable cell count, active SH groups, total volatile fatty acid, free fatty acid, electrophoretic patterns of degraded caseins, and viscosity were analyzed to investigate physicochemical properties of fermented slurries. Acid production was accelerated in the cheese slurries with protease than that without the enzyme and PH of the former was decreased after three days of fermentation to 4.90. The Change of titratable acidity agreed to PH patterns. Soluble nitrogen of the Control slurry was increased slowly for four days and then rapidly to 40% of total nitrogen while those containing protease to 70%. The protease of lactic cultures used (Streptococcus lactis and Streptococcus cremoris) broke down as-casein more rapidly than $\beta$-casein and most proteins were degraded to peptides and amino acids after three days of fermentation. Total volatile fatty acids were increased by added lipase and free fatty acids composition analyzed by GLC in cheddar slurry with 0.00001% lipase was similar to that of commercial cheddar cheese, while that in Italian-type slurry was a half of that in commercial Italian cheese. Active SH groups were increased in the cheese slurries with glutathione from fourth day of fermentation. The viscosity of slurries decreased very rapidly by addition of protease.

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Pharmacological Activity of Chaga Mushroom on Extraction Conditions and Immunostimulating Polysaccharide (추출조건에 따른 차가버섯 생리활성 및 면역활성 다당)

  • Baek, Gil-Hun;Jeong, Heon-Sang;Kim, Hoon;Yoon, Taek-Joon;Suh, Hyung-Joo;Yu, Kwang-Won
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.41 no.10
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    • pp.1378-1387
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    • 2012
  • To investigate the pharmacological activity of chaga mushroom (Inonotus obliquus) on extraction conditions, chaga was extracted using water (reflux at $50^{\circ}C$, decoction over $90^{\circ}C$, pressure at $121^{\circ}C$) or ethanol (reflux at 50, 70, or $90^{\circ}C$). When water extract was further fractionated into crude polysaccharide (IO-CP), yields of IO-CP (4.8~16.8%) were higher than those of ethanolic extracts (IO-E, 1.9~2.7%) at increased temperature. For antioxidant activity, crude polysaccharide (IO-CP-121) obtained by pressurized extraction showed the highest polyphenolic and flavonoid contents (35.10 mg TAE/g and 18.48 mg QE/g, respectively) as well as DPPH and ABTS free radical scavenging activities (26.08 and 27.99 mg AEAC/100 mg, respectively). Meanwhile, IO-CP-D (decoction) and IO-CP-50 (reflux) had more potent mitogenic effects (2.10- and 1.95-fold of saline control at 100 ${\mu}g/mL$) as well as intestinal immune system modulating activities (6.30- and 5.74-fold) compared to IO-CP-121, whereas ethanolic extracts showed no activity. Although no IO-CP showed cytotoxicity against RAW 264.7 cells at 0.1 mg/mL, IO-CP-121 significantly inhibited TNF-${\alpha}$ and NO production as pro-inflammatory factors in LPS-stimulated RAW 264.7 cells (29.2 and 63.5%, respectively). Ethanolic extracts also showed no cytotoxicity at 0.1 mg/mL, whereas inhibition of TNF-${\alpha}$ and NO production was significantly low compared to that of IO-CP-121. In addition, active IO-CP-D was further fractionated into an unadsorbed (IO-CP-I) and seven adsorbed fractions (IO-CP-II~VIII) by DEAE-Sepharose CL-6B column chromatography in order to isolate immunostimulating polysaccharide. IO-CP-II showed the most potent mitogenic effect and macrophage stimulating activity (4.51- and 1.64-fold, respectively). IO-CP-II mainly contained neutral sugars (61.86%) in addition to a small amount of uronic acid (2.96%), and component sugar analysis showed that IO-CP-II consisted mainly of Glc, Gal, and Man (molar ratio of 1.00:0.55:0.31). Therefore, extraction conditions affect the physiological activity of chaga, and immunostimulating polysaccharide fractionated from chaga by decoction is composed mainly of neutral sugars.

Immune Enhancement of Polysaccharide from Submerged Culture with Phellinus linteus in the Medium Supplemented with Ginseng Extract (수삼추출물 첨가 혼합배지에서 조제된 상황 균사체 심부배양물 다당획분의 면역활성 증진)

  • Kim, Hoon;Song, Ki-Yun;Jeong, Jae-Hyun;Jeong, Heon-Sang;Lee, Hyeon-Yong;Yu, Kwang-Won
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.40 no.1
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    • pp.20-28
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    • 2011
  • Crude polysaccharide (CP) was fractionated from the submerged culture (containing both mycelia and culture broth, SC) with Phellinus linteus (PL) in mushroom complete medium (MCM) supplemented with ginseng extract ($65^{\circ}$Bx, GE) to enhance the immune activity. PL-GE-15-CP from SC cultivated in MCM supplemented with GE-15% (v/v, a ratio of MCM volume to GE) showed significantly higher macrophage stimulation (1.45 fold of the saline control at $100{\mu}g$/mL) than PL-GE-5 and 10-CP with GE-5 and 10%, or PL-CP from SC without GE. The potent intestinal immune system modulating activity through Peyer's patch was also obtained by PL-GE-15-CP (1.46 fold). When PL-GE-15-CP further fractionated on DEAE-Sepharose CL-6B (Cl- form), PL-GE-15-CP-II was the significantly higher than others from PL-GE-15-CP or PL-CP on macrophage stimulation, interleukin (IL)-12 production and intestinal immune system modulation (1.54, 3.96 and 1.56 fold, respectively). PL-GE-15-CP-II also had higher anti-metastatic activity against colon 26-M3.1 carcinoma cell (57.3% inhibition of tumor control, $200{\mu}g$/mouse) rather than PL-CP-II. This active fraction (PL-GE-15-CP-II) mainly contained neutral sugar (82.45%) and uronic acid (12.99%), and component sugar analysis showed that PL-GE-15-CP-II consisted mainly of uronic acid, Ara, Man, Gal and Glc (molar ratio of 0.52:0.97:0.63:1.00:0.54). Furthermore, the activity of GE culture was higher compared with culture without GE, indicating that GE helped to enhance the immune activity of P. linteus; also, it is assumed that the polysaccharide plays an important role in immune enhancement.

Lipid Composition of Improved Rape Seed on the Content of Erucic acid (에루신산에 대(對)한 개량(改良) 유채종자(油菜種子)의 지질구성(脂質構成) 성분(成分))

  • Lee, Sung-Ho;Kim, Chung-Ki;Kim, Hong-Su
    • Applied Biological Chemistry
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    • v.28 no.4
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    • pp.245-251
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    • 1985
  • In order to analyze the lipid composition ana chemical characteristics of rapeseed oil in relation to content of erucic acid, three recommendable cultivars(MOK-PO 70, MOK-PO DANGYO 12 and 14) seeds were used. The composition of lipid was confirmed by column chromatography and TLC, and the composition of fatty acid in total and neutral lipid was determined by GLC, and the results were as follows. 1. The crude fat was 44.0% to 45.3% in all cultivars. 2. The iodine value, saponification value, acid value and nonsaponifiable content were 100 to 101, 176 to 184, 0.7 to 0.9 and 1.0 to 1.2, respectively, with no remarkable difference among three cultivars. 3. The content of neutral lipid was 95.3% to 96.3% of total lipid, and others were compound lipids. 4. The content to triglyceride was 92. 1% to 92.5% of total lipid. Diglyceride and sterol ester were contained 1.1% and 1.2%, respectively. There was a small quantity of free fatty acid and free sterol and monoglyceride was little. 5. The erucic acid was the highest (26.4%), in MOK-PO DANGYO 12 among the composition of fatty acid, while those of MOK-PO 70 and MOK-PO DANGYO 14 were so small as 1.0% and 3.9%. The content of erucic acid was decreased with the increase of oleic acid, the content of oleic acid in MOK-PO DANGYO 12 was 30%, 63% in MOK-PO 70 and, 60% in MOK-PO DANGYO 14. The content of linoleic acid and linolenic acid showed no remarkable difference but that of eicosenoic acid was 11.7% in MOK-PO DANGYO 12 as the highest. The ratio of the fatty acid content in eluted neutral lipid and the total oil showed no remarkable differences.

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Some Chemical Properties and Composition of Lipid Extracts of Riced Dehydrated Potato Granules (감자분말(粉末)의 지방질조성(脂肪質組成)과 저장중(貯藏中)의 변화(變化)에 관(關한) 연구(硏究))

  • Yoon, Jeong-Won;Hong, Bum-Shik;Yang, Han-Chul;Kim, Dong-Hoon
    • Korean Journal of Food Science and Technology
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    • v.10 no.3
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    • pp.320-330
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    • 1978
  • Riced dehydrated potato granules with a good reconstituton quality was made from Irish Cobber (IC) and Shimabara which are representative varieties in Korea. A part of IC potato granules was packed in small vinyl-bags while the rest, including some BHA (75ppm)-treated granules, was packed in $301{\times}407$ plain tin cans. These granules were stored in a room at room temperature, and their physico-chemical properties and lipid composition were studied. The color of the granules was measured with a Hunter-type Tristimules colorimeter. L, $a_L,$ and $b_L-values$ were respectively 83.8, -1.1 and 18.3. Variety and package-type did not affect the color. In general, it was darker than that of commercial wheat flour, but whiter than that of commercial defatted soy flour. At $100^{\circ}C$, the granules, irrespective of variety and package-type, absorbed water rapidly, and reached the maximum moisture content of $90{\sim}92%$ in 3 min., whereas they absorbed water more slowly at $8{\sim}14^{\circ}C$, and reached the maximum content of $72{\sim}74%$ in 5 minutes. Peroxide, TBA, carbonyl, acid, and iodine values of the granules after 3 months storage were respetively $150{\sim}460\;meq/kg,\;20{\sim}26,\;154{\sim}380$ micromole, $24{\sim}59,\;and\;70{\sim}78$. Except iodine values, all the chemical values were affected by variety and package-type. Ether and $CHCl_3-extracts$ of the granules from IC and Shimabara were subjected to GLC and TLC analyses. In case of IC, the major fatty acids were palmitic, stearic, and oleic (30.0, 18.8, 40.6), while in case of Shimabara they were palmitic, oleic, and linoleic acids (26.7, 39.6, 23.4%). The major lipid classes were, in both cases, triglycerides and phospholipids. Their contents were raspectively 19.1 and 43.1 (IC), and 30.1 and 37.4% (Shimabara).

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Solution Phase Photolyses of Substituted Diphenyl Ether Herbicides under Simulated Environmental Conditions (모조(模造) 환경조건하(環境條件下)에서의 치환(置換) Diphenyl Ether 제초제(除草劑)의 광분해(光分解)에 관(關)한 연구(硏究))

  • Lee, Jae-Koo
    • Applied Biological Chemistry
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    • v.17 no.3
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    • pp.149-176
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    • 1974
  • Eight substituted diphenyl ether herbicides and some of their photoproducts were studied in terms of solution phase photolysis under simulated environmental conditions by using a Rayonet photochemical reactor. The test compounds absorbed sufficient light energy at the wavelength of 300 nm to undergo various photoreactions. All the photoproducts were confirmed by means of tlc, glc, ir, ms, and/or nmr spectrometry. The results obtained are summarized as follows: Solution phase photolysis of C-6989: An exceedingly large amount of p-nitrophenol formed strongly indicates the readiness of the ether linkage cleavage of this compound as the main reaction in all solvents used. Photoreduction of nitro to amino group(s) and photooxidation of trifluoromethyl to carboxyl group were recognized as minor reactions. Aqueous photolysis of p-nitrophenol: Quinone(0.28%), hydroquinone (0.66%), and p-aminophenol (0.42%) were confirmed as photoproducts, in addition to a relatively small amount of an unknown compound. The mechanisms of formation of these products were proposed to be the nitro-nitrite rearrangement via $n{\rightarrow}{\pi}^*$ excitation and the photoreduction through hydrogen abstractions by radicals, respectively. Solution phase photolysis of Nitrofen: Photochemical reduction leading to the p-amino derivative was the main reaction in n-hexane. In aqueous solution, the photoreduction of nitro to amino group and hydroxylation predominated over the ether linkage cleavage. Nucleophilic displacement of the nitro group by hydroxide ion and replacement of chlorine substituents by hydroxyl group or, to a lesser extent, hydrogen were also observed as minor reactoins. Solution phase photolysis of MO-338: Photoreduction of the nitro to amino group was marked in the n-hexane solution photolysis. In the aqueous solution, photoreduction of the nitro substituent and hydroxylation were the main reactions with replacement of chlorine substituents by the hydroxyl group and hydrogen, and cleavage of the ether linkage as minor reactions. Photolyses of MC-4379, MC-3761, MC-5127, MC-6063, and MC-7181 in n-hexane and cyclohexane: Photoreduction of the nitro group leading to the corresponding amino derivative and replacement of one of the halogen substituents by hydrogen from the solvent used were the key reactions in each compound. Aqueous photolysis of MC-4379: Cleavage of the ether linkage, replacement of the carboxymethyl by hydroxyl group, hydroxylation, and replacement of the nitro by hydroxy group were prominent with photoreduction and dechlorination as minor reactions. Aqueous photolysis of MC-3761: Cleavage of the ether linkage, replacement of the carboxymethyl by hydroxyl group, and photoreduction followed by hydroxylation were the main reactions. Aqueous photolysis of MC-5127: Replacement of carboxyethyl by hydrogen was predominant with ether linkage cleavage, photoreduction, and dechlorination as minor reactions. It was obvious that the decarboxyethylation proceeded more readily than decarboxymethylation occurring in the other compounds. Aqueous photolysis of MC-6063: Cleavage of the ether linkage and photodechlorination were the main reactions. Aqueous photolysis of MC-7181: Replacement of the carboxymethyl group by hydrogen and monodechlorination were the remarkable reactions. Cleavage of the ether linkage and hydroxylation were thought to be the minor reactions. Aqueous photolysis of 3-carboxymethyl-4-nitrophenol: The photo-induced Fries rearrangement common to aromatic esters did not appear to occur in the carboxymethyl group of this type of compound. Conversion of nitro to nitroso group was the main reaction.

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