• Natural Product Sciences
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• v.5 no.2
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• pp.80-84
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• 1999

Peroxynitrite is a cytotoxic intermediate produced by the reaction between the superoxide anion radical and nitric oxide. Flavonoids (afzelin, quercitrin and quercetin 3-O-sambubioside), and chlorogenic acid and its methyl ester obtained from leaves of loquat (Eriobotrya japonica) have recently been shown to scavenge 1,1-diphenyl-2-picrylhydrazyl radical and to inhibit lipid peroxidation in mouse liver homogenate. The aim of this study is to investigate the inhibitory effects of the above components on peroxynitrite produced stimulated by 3-morpholinosydnonimine (SIN-1) to produce superoxide anion radical and nitric oxide at the same time. In addition, the present study tests whether or not the components directly scavenge peroxynitrite itself. The results showed that the components with the aromatic ortho-dihydroxyl groups (catechol) were more potent inhibitors of peroxynitrite formation by SIN-1. In particular, the methyl ester form of chlorogenic acid showed the most potent inhibition. At $5\;{\mu}M$ concentration, the order of minimizing peroxynitrite formation were : methyl chlorogenic acid > quercitrin > quercetin 3-O-sambubioside > chlorogenic acid > afzelin. Authentic peroxynitrite was directly scavenged by the components in a manner similar to peroxynitrite formation by SIN-1. In particular, when compared with penicillamine as a positive control, methyl chlorogenate was as effective in inhibiting peroxynitrite formation and approximately 2 times more effective in scavenging an authentic peroxynitrite. These results demonstrate therefore, that components extracted from the leaves of Eriobotrya japonica effectively scavenged peroxynitrite.

• Pediatric Gastroenterology, Hepatology & Nutrition
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• v.18 no.2
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• pp.121-127
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• 2015

Purpose: To evaluate clinical and laboratory profile of Wilson's disease (WD) in children. Methods: This cross sectional study was conducted at Bangabandhu Sheikh Mujib Medical University Hospital. Bangladesh, over a period of 3 years. One hundred consecutive children of WD between 3 to 18 years of age were evaluated. Results: Mean age was $8.5{\pm}1.5years$. Male female ratio was 2:1. Ninety-one percent of patients were Muslim and 9% Hindu. A total of 53% cases of hepatic WD presented between 5 to 10 years of age and most of the neurologic WD manifested in 10-15 years age group. Sixty-nine children presented only with hepatic manifestations, 6 only with neurological manifestations, 14 with both hepatic and neurological manifestation, 10 children was asymptomatic and 1 patient presented with psychiatric features. WD presented as chronic liver disease (CLD) in 42%, CLD with portal hypertension in 34%, acute hepatitis in 20% and fulminant hepatic failure in 4% cases. Stigmata of CLD were found in 18% patients. Keiser-Fleischser ring was found in 76% total patients. Elevated serum transaminase was found in 85% cases, prolonged prothrombin time in 59% cases and hypoalbuminaemia in 53% cases. A total of 73% patients had low serum ceruloplasmin, basal urinary copper of >$100{\mu}g/day$ was found in 81% cases and urinary copper following penicillamine challenge of >$1,200{\mu}g/day$ was found in 92% cases. Conclusion: Majority of studied WD children presented with hepatic manifestation of which 76% presented with CLD. Any child presented with jaundice after the age of 3 years should be investigated for WD.

• Journal of Animal Reproduction and Biotechnology
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• v.35 no.2
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• pp.142-148
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• 2020

The research work was undertaken to determine an effective fertilization medium, sperm separation method and sperm capacitating agent for optimum in vitro fertilization (IVF) rates of indigenous zebu cow oocytes. In experiment 1, tissue culture medium (TCM 199), Tyrode's albumin lactate pyruvate (TALP) and Brackett and Oliphant (BO) medium were used as basic medium for IVF of oocytes of indigenous zebu cows. In experiment 2, three sperm separation methods namely centrifugation, swim up and percoll gradient methods were used for separation of motile and viable spermatozoa for IVF. In experiment 3, for capacitation of spermatozoa, IVF medium supplemented with the heparin, mixture of penicillamine, hypotaurine and epinephrine (PHE) or the combination of heparin with PHE were used for fertilization. In vitro culture (IVC) of presumptive zygotes was done in modified synthetic oviduct fluid (mSOF) medium using standard procedure 24 h after sperm-oocytes co-culture. The cleavage rate was determined to evaluate the efficacy of fertilization medium, sperm separation method and sperm capacitating agent 24 h after IVC. The cleavage rate was higher in oocytes fertilized in TALP (63.3%) than in TCM 199 (47.5%) (p < 0.05). The cleavage rate was higher in oocytes fertilized by spermatozoa separated by percoll gradient method (62.3%) than by centrifugation (51.6%) (p < 0.05). The cleavage rate of oocytes was higher when insemination was done with spermatozoa capacitated in TALP supplemented with heparin and PHE (61.3%) compared to control (40.9%) (p < 0.05). In conclusions, TALP based medium and percoll gradient sperm separation followed by capacitation with combination of heparin and PHE are suitable for IVF of indigenous zebu cow oocytes in Bangladesh.

• Journal of Preventive Medicine and Public Health
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• v.7 no.2
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• pp.377-381
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• 1974

Determination of lead in urine is important in industrial hygiene and toxicology. Dithizone method has been principally used for the determination of lead in urine, which gives accurate results in skilful hands but is usually complex and time-consuming. Atomic absorption spectrophotometry is a new simple method and several procedures have been described. However, the influences of pH and the presence of chelating agents during treatment of lead poisoning are not clear. The purpose of this study was to find out the effect of pH and chelating agents on the determination of lead using Shimadzu atomic absorption/flame spectrophotometer, model AA-610. The results obtained were as follows: 1. The atomic absorption spectrophotometry(AAS) could be applied without prior acid digestion to specimens in the absence of chelating agents. The absorbance at $2,170\;{\AA}$, though more sensitive, was more noisy electronically. Therefore, we selected the wavelength of $2,833\;{\AA}$ plus scale expansion. 2. The optimal pH was in the range from 2 to 3. 3. The sensitivity was $0.075{\mu}g/ml/%$ and detection limit was about $0.2{\mu}g/ml$. 4. In the presence of EDTA, lead could not be completely determined without prior acid digestion. 5. On specimens from patients receiving penicillamine therapy, a comparison was made between the values obtained with dithizone method and AAS method with prior acid digestion. The results of comparison showed a very good agreement.

• Archives of Pharmacal Research
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• v.29 no.12
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• pp.1074-1079
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• 2006

As part of our research on phytochemicals that exert protective effects against diseases related to reactive nitrogen species, we have evaluated the scavenging activity of the yellow leaves of Ginkgo biloba on $ONOO^{-}$. The methanol extract and ethyl acetate fraction obtained from yellow leaves of G. biloba evidenced a marked scavenging activity on authentic $ONOO^{-}$. Repeated column chromatography of the active ethyl acetate soluble fraction on silica gel, Sephadex LH-20, and RP-18, resulted in the purification of 15 known compounds, including sciadopitysin (1), ginkgolide B (2), bilobalide (3), isoginkgetin (4), kaempferol (5), luteolin (6), protocatechuic acid (7), bilobetin (8), amentoflavone (9), ${\beta}-sitosterol$ glucopyranoside (10), kaempferol 3-O-rhamnopyranoside (11), kaempferol 3-O-glucopyranoside (12), kaempferol $3-O-[{6^{'}-O-p-coumaroyl-{\beta}-D-glucopyranosyl(1{\rightarrow}2)-{\alpha}-L-rhamnopyranoside]$ (13), kaempferol 3-O-rutinoside (14), and 6-hydroxykynurenic acid (15). Among the compounds isolated, flavonoids (5, 6 and 11-14), protocatechuic acid (7), and 6-hydroxykynurenic acid (15) all exhibited marked scavenging activities on authentic $ONOO^{-}$. The $IC_{50}$ values of 5-7, 11-14 and 15 were as follows: $2.86{\pm}0.70,\;2.30{\pm}0.04,\;2.85{\pm}0.10,\;5.60{\pm}0.47,\;4.16{\pm}1.65,\;2.47{\pm}0.15,\;3.02{\pm}0.48,\;and\;6.24{\pm}0.27\;{\mu}M$, respectively. DL-Penicillamine ($IC_{50}=4.98{\pm}0.27\;{\mu}M$) was utilized as a positive control. However, the other compounds (1-4, 8-10) exerted no effects against $ONOO^{-}$.

• BMB Reports
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• v.41 no.2
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• pp.164-169
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• 2008

The tumor suppressor gene p53 regulates apoptotic cell death and the cell cycle. In this study, we investigated the role of p53 in nitric oxide (NO)-induced apoptosis in vascular smooth muscle cells (VSMCs). We found that the NO donor S-nitroso-N-acetyl-penicillamine (SNAP) increased apoptotic cell death in p53-deficient VSMCs compared with wild-type cells. The heme oxygen-ase (HO) inhibitor tin protoporphyrin IX reduced the resistance of wild-type VSMCs to SNAP-induced cell death. SNAP promoted HO-1 expression in both cell types. HO-2 protein was increased only in wild-type VSMCs following SNAP treatment; however, similar levels of HO-2 mRNA were detected in both cell types. SNAP significantly increased the levels of non-heme-iron and dinitrosyl iron-sulfur clusters in wild-type VSMCs compared with p53-deficient VSMCs. Moreover, pretreatment with FeSO4 and the carbon monoxide donor CORM-2, but not biliverdin, significantly protected p53-deficient cells from SNAP-induced cell death compared with normal cells. These results suggest that wild-type VSMCs are more resistant to NO-mediated apoptosis than p53-deficient VSMCs through p53-dependent up-regulation of HO-2.

• Natural Product Sciences
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• v.9 no.2
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• pp.73-79
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• 2003

To discover the sources with antioxidative activity in traditional medicines, 100 extracts of Korean medicinal plants were screened for their scavenging effect on peroxynitrite $(ONOO^{-})$ and total reactive oxygen species (ROS). The potency of total ROS scavenging activity was shown in the extracts of 25 plants, and 4 of their species, Macleaya cordata R. Br., Salvia plebeia R. Br., Cassia tora L. and Angelica gigas Nakai, had a greater effect with $IC_{50}$ values of $1.7{\pm}0.36$, $4.3{\pm}1.08$, $4.9{\pm}0.17$ and $5.8{\pm}1.01\;{\mu}g/ml$, respectively, than that of trolox, positive control $(7.61{\pm}0.12\;{\mu}g/ml)$. Another 35 extracts exhibited inhibitory effect of below 50 percent at $100\;{\mu}g/ml$ of sample concentrations on total ROS, while the rest observed total ROS generators rather than scavengers. The peroxynitrite scavenging activities were observed in the greater part of the plants tested. Five of them, Schisandra chinensis Baill, Campsis grandiflora (Thunb.) K. Schum., Cedrela sinensis A. Juss., Pleuropterus multiflorus Turcz. and Veronica linariaefolia Pall represented scavenging activities on peroxynitrite twice as strong with $IC_{50}$ Values of $0.48{\pm}0.10$, $0.59{\pm}0.15$, $0.60{\pm}0.10$, $0.64{\pm}0.10$ and $0.91{\pm}0.23\;{\mu}g/ml$, respectively, as that of penicillamine $(1.72{\pm}0.05\;{\mu}g/ml)$, positive control. Consequently, 25 species of the entire plants tested, exhibited scavenging activities on total ROS and $ONOO^{-}$, Salvia plebeia R. Br., Macleaya cordata R. Br., Cassia tora L. and Angelica gigas Nakai exerted potent scavenging activities on both radicals.

• Natural Product Sciences
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• v.14 no.1
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• pp.1-11
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• 2008

Peroxynitrite ($(ONOO^-)$ is a cytotoxic species formed from nitric oxide and superoxide anion, which are highly implicated in the pathogenesis of oxidative stress-mediated diseases. The aim of this study was to investigate the scavenging effects of Phellinus linteus on authentic $ONOO^-$, and further phytochemical studies are planned that will attempt to identify the active principles. From the active EtOAc fraction, a mixture of fungisterol and 5-dihydroergosterol (1), a mixture of betulin and 1,2-benzenedicarboxylic acid bis (2-methyl heptyl) ester (2), protocatechualdehyde (3), protocatechuic acid (4), cirsiumaldehyde (5), hispidin (6), caffeic acid (7), phelligridin D (8), uracil (9), gallic acid (10), 2,5-dihydroxybenzoic acid (11), ferulic acid (12), 2,3-dihydroxybenzaldehyde (13), arbutin (14), isoferulic acid (15), guanosine (16), and ellagic acid (17) were isolated, and their structures were characterized based on spectroscopic data. All compounds except 3, 6, 7 and 16 were isolated for the first time from P. linteus. Compounds 3, 4, 6-8, 10-15, and 17 showed potent scavenging activity on $ONOO^-$, with $IC_{50}$ values of $2.06\;{\pm}\;0.10$, $3.45\;{\pm}\;0.57$, $0.71\;{\pm}\;0.05$, $2.78\;{\pm}\;0.36$, $5.42\;{\pm}\;0.26$, $1.13\;{\pm}\;0.02$, $1.82\;{\pm}\;0.17$, $0.91\;{\pm}\;0.19$, $1.59\;{\pm}\;0.09$, $1.88\;{\pm}\;0.07$, $1.22\;{\pm}\;0.37$, and $2.01\;{\pm}\;0.02\;{\mu}M$, respectively, as compared to the positive control, DL-penicillamine, with an $IC_{50}$ value of $5.04\;{\pm}\;0.06\;{\mu}M$.

• Archives of Pharmacal Research
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• v.28 no.5
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• pp.534-540
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• 2005

Activity-guided fractionation of the CH$_2Cl_2$-soluble fraction of the roots of Sophora flavescens furnished five 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scaveng ers: trans-hexadecyl ferulic acid (1) cis-octadecyl ferulic acid (2), trans-hexadecyl sinapic acid (3), (-)-4-hydroxy-3-methoxy-(6aR,11aR)-8, 9-methylenedioxypterocarpan (4) and desmethylanhydroicaritin (8), along with nine known inactive compounds: (-)-maackiain (5), xanthohumol (6), formononetin (7), (2S)-2'-methoxykurarinone (9), (2S)-3${\beta}$,7,4'-trihydroxy-5-methoxy-8-(${\gamma},{\gamma}$- imethylallyl )-flavanone (10), (2S)-7,4'-dihydroxy-5-methoxy-8- (${\gamma},{\gamma}$-dimethylallyl ) -flavanone (11), umbelliferone (12), kuraridin (13), and trifolirhizin (14). Compounds 1-4 and 8 exhibited DPPH free radical scavenging effects at IC$_{50}$ values of 33.01 ${\pm}$ 0.20, 57.06 ${\pm}$ 0.16, 39.84 ${\pm}$ 0.36, 35.83 ${\pm}$ 0.47, and 18.11 ${\pm}$ 0.04${\mu}$M, respectively. L-Ascorbic acid, when used as a positive control, exhibited an IC$_{50}$ value of 7.39 ${\pm}$ 0.01 ${\mu}$M. Compounds 1-4 and 8 also appeared to exert significant scavenging effects on authentic ONOO-, with IC$_{50}$ values of 5.76 ${\pm}$ 1.19, 15.06 ${\pm}$ 1.64, 8.17 ${\pm}$ 4.97, 1.95 ${\pm}$ 0.29 and 4.06 ${\pm}$ 2.41 ${\mu}$M, respectively. Penicillamine (IC$_{50}$= 2.36 ${\pm}$ 0.79${\mu}$M) was used as a positive control. In addition, compounds 2,4,6,8, and 10 were isolated from this plant for the first time.

• Korean Journal of Plant Resources
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• v.29 no.1
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• pp.20-25
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• 2016

Five caffeoylquinic acids of Aster altaicus var. uchiyamae Kitamura (Compositae) leaves were identified using standard compounds by HPLC and determined as follows: 3,4-di-O-caffeoylquinic acid (4.92 ± 0.06 ㎎/g dried weight), 3,5-di-O-caffeoylquinic acid (3.95 ± 0.13 ㎎/g), 4,5-di-O-caffeoylquinic acid (1.39 ± 0.10 ㎎/g), 5-O-caffeoylquinic acid (chlorogenic acid, 8.05 ± 0.21 ㎎/g), 3-O-caffeoylquinic acid (4.97 ± 0.18 ㎎/g). The total content of five caffeoylquinic acids were calculated as 26.73 ± 0.26 ㎎/g dried weight while the percentage of the five compounds in the MeOH extract was calculated as 25.22 ± 0.25%. The IC₅₀ value of the MeOH extract scavenging peroxynitrite (ONOO⁻) was shown as 5.16 ± 0.15 ㎍/㎖.