Application of polyethyleneimine cellulose for the class separation of steroidal carboxylic acids from neutral steroids and pigments in urine

Metabolites of corticosteroids that contain the 21-oic acid moiety are found in human urine. The acids from neutral steroids and urinary pigments have been separated by passing the mixture through a column of polyethyleneimine cellulose. The acids adhering to the column are quantitatively eluted with dilute formic acid. The purified preparation is suitable for derivatization and chromatographic analysis.     

Identification of 5-desmethylubiquinone-9 as an intermediate in the biosynthesis of ubiquinone-9 by rat liver mitochondria

Radioactive 5-desmethylubiquinone-9 has been isolated from mitochondria synthesizing ubiquinone-9-¹⁴C from p-hydroxybenzoate-U-¹⁴C. By mass spectrometry, the natural 5-desmethylubiquinone-9 has been shown to be identical with that chemically synthesized from fumigatol and solanesol. Synthetic 5-desmethylubiquinone-9-³H can be methylated to ubiquinone-9-³H by S-adenosyl-L-methionine in submitochondrial particles.     

Characterization of B and H blood-group active glycosphingolipids from human B erythrocyte membranes

Two blood group B active glycosphingolipids (B-I and B-II) previously isolated and highly purified from human B erythrocytes [21] were analysed first by degradation with α-D-galactosidase from coffee beans, α-L-fucosidase from bovine kidney and with 0,1 N trichloracetic acid; the native B-glycolipids as well as their degradation products were then investigated by methylation analysis with combined gas chromatography-mass spectrometry, by thin layer chromatography, twodimensional immunodiffusion and by the hemagglutination inhibition technique. Together with the results obtained by mass spectrometry of permethylated glycolipids [26] the following structures were elucidated: α-D-galactopyranosyl-(1 → 3)-[α-L-fucopyranosyl-(1 → 2)]-D-galactopyranosyl-(1 → 4)-N-acetyl-D-glucosaminosyl-(1 → 3)-D-galactopyranosyl-(1 → 4)-D-glucopyranosyl-(1 → 1)-ceramide for the B-I glycosphingolipid and α-D-galactopyranosyl-(1 → 3)-[α-L-fucopyranosyl-(1 → 2)]-D-galactopyranosyl-(1 → 4)-N-acetyl-D-glucosaminosyl-(1 → 3)-D-galactopyranosyl-(1 → 4)-N-acetyl-D-glucosaminosyl-(1 → 3)-D-galactopyranosyl-(1 → 4)-D-glucopyranosyl-(1 → 1)-ceramide for the B-II glycosphingolipid. A H active glycolipid fraction from B erythrocytes further purified by thin layer chromatography was also investigated by methylation analysis. The pattern of its partially methylated alditol acetates was essentially the same as that of the α-galactosidase treated and permethylated B-I glycolipid. It also exhibited strongly precipitating and hemagglutination inhibiting H properties as well as the two α-galactosidase treated B-I and B-II glycosphingolipids. Based upon these data the following tentative structure was proposed: α-L-fucopyranosyl-(1 → 2)-D-galactopyranosyl-(1 → 4)-N-acetyl-D-glucosaminosyl-(1 → 3)-D-galactopyranosyl-(1 → 4)-D-glucopyranosyl-(1 → 1)-ceramide. Gas chromatographic analysis revealed sphingosine and lignoceric, nervonic and behenic acids to be the main components of the ceramide residues of the three glycosphingolipids. From the data presented the H active substance very probably can be regarded as the immediate precursor of the B-I glycosphingolipid from human B erythrocyte membranes.     

Dehydrophylloquinone, α-dehydrotocopherolquinone and dehydrotocopherols from etiolated maize and barley shoots

Dehydrophylloquinone, α-dehydrotocopherolquinone, α-dehydrotocopherol and γ-dehydrotocopherol have been isolated from etiolated maize and barley shoots and excised etiolated maize shoots that have been exposed to light. They are not present, however, in green maize shoots, spinach leaves and etiolated bean leaves. Demethylphylloquinone was not detected in any of the tissues analysed.     

16SrRNA sequencing of Dye decolorizing bacteria isolated from Soil

Dye׳s residues in textile effluents are hazardous for humans and animals health. Such pollutants can be degraded into non-harmful molecules using biological approaches that are considered cheaper and ecologically safer. Isolated 15 bacterial cultures from soil that could be used in biological system were showed decolorization capacity for Acid Green dye (33.9% to 94.0%) using thin layer chromatography and broth culture method. The most promising cultures (AMC3) to decolorize Acid green Dye (94.6%) was re-coded as NSDSUAM for submitting at IMTECH, Chandigarh for sequencing. The 16SrRNA sequencing suggested that it can be a variant of Pseudomonas geniculata (99.85% identical similarity) with difference of 2 base pairs to reference strain Pseudomonas geniculata ATCC 19374(T). Thus present study proposed dye decolorizing efficiency of the isolated strain of Pseudomonas geniculata that was previously unnoticed. The sequence is deposited in NCBI GenBank with the accession number {"type":"entrez-nucleotide","attrs":{"text":"KP238100","term_id":"745856746","term_text":"KP238100"}}KP238100.     

Validated TLC method for simultaneous quantitation of kutkoside and picroside‐I from Kutki extract

Introduction – The two iridoid glycosides kutkoside and picroside‐I are the active hepatoprotective principles of Picrorhiza kurroa Royle ex Benth (Scrophulariaceae), commonly known as Kutki. Quantitation of these phytoconstituents is important for the routine quality control of Kutki extract. Objective – To develop and validate a simple, precise and rapid thin‐layer chromatography (TLC) method for the simultaneous quantitation of kutkoside and picroside‐I in Kutki extract. Methodology – The analysis was performed on a TLC precoated silica gel 60 F₂₅₄ plate with ethyl acetate:methanol:glacial acetic acid:formic acid (25:5:1:1, v/v/v/v) as mobile phase. Densitometric evaluation of kutkoside and picroside‐I was carried out at 265 nm and the mobile phase showed good resolution with R_f values 0.42 ± 0.03 and 0.61 ± 0.03 for kutkoside and picroside‐I, respectively. The method was validated in terms of specificity, linearity, accuracy and precision. Results – The content of kutkoside and picroside‐I was found to be 2.18 and 1.90%, respectively, and was comparable with those obtained by HPLC. The linearity was found to be in the range of 80–480 ng/spot for both kutkoside and picroside‐I. The average recovery values were found to be 96.5 and 96.0% for kutkoside and picroside‐I, respectively. Conclusion – The developed method was found to be relatively simple, precise and reproducible for the simultaneous quantitation of kutkoside and picroside‐I. The method does not employ any derivatisation procedure and can be used as a quality control tool for the routine analysis of commercial Kutki extracts. Copyright © 2010 John Wiley & Sons, Ltd.     

肾衰康浓缩丸质量标准研究

目的：建立肾衰康浓缩丸质量控制方法。方法：采用TLC鉴别肾衰康浓缩丸中大黄、丹参;采用HPLC-ELSD对制剂中黄芪甲苷进行含量测定,流动相为乙腈-水（30：70）,流速：1 mL/min,柱温：28℃,漂移管温度：42℃,氮气：355kPa。结果：大黄、丹参的TLC鉴别斑点清晰、专属性强;黄芪甲苷在1.0μg-10.0μg之间呈良好的线性关系（r=0.9997,n=5）,平均回收率为99.81%,RSD为1.73%（n=6）。结论：所建立的方法简便准确、专属性强、重现性好,可作为控制肾衰康浓缩丸质量的方法。     

Bioassay guided fractionation of bioactive metabolite from Corynascus verrucosus inhabiting Croton bonplandianus Baill

Mycoendophytes have been a promising source of bioactive metabolites for new developments in pharmaceuticals and industrial relevance. During the screening of mycoendophytes diversity from C. bonplandianus, bioactive C. verrucosus CBF 22 was isolated from the leaf. The bioactive isolate was subjected to fermentation and the ethyl acetate fraction was evaluated for antimicrobial activity by disc diffusion and microbroth dilution methods, followed by cytotoxicity using MTT assay. The results revealed significant activity against Gram negative bacteria, Gram positive bacteria, and human pathogenic dermatophytic fungi, C. albicans. The TLC Bioautography-Guided approach employed for the detection of antimicrobial metabolite from bioactive fraction was depicted by a zone of inhibition. Antimicrobial screening of mycoendophytes C. bonplandianus could envision a unique potential niche for bioactive compounds of pharmaceutical importance. These findings suggest C. verrucosus as a potent candidate for producing natural bioactives, to combat multidrug-resistant pathogens. This work is the first report on the incidence of C. verrucosus as mycoendophyte and its bioactive potential of inhabiting C. bonplandianus Baill.     

Understanding the muscular dystrophy caused by deletion of choline kinase beta in mice

Choline kinase in mice is encoded by two genes, Chka and Chkb. Disruption of murine Chka leads to embryonic lethality, whereas a spontaneously occurring genomic deletion in murine Chkb results in neonatal bone deformity and hindlimb muscular dystrophy. We have investigated the mechanism by which a lack of choline kinase β, encoded by Chkb, causes hindlimb muscular dystrophy. The biosynthesis of phosphatidylcholine (PC) is impaired in the hindlimbs of Chkb^−/− mice, with an accumulation of choline and decreased amount of phosphocholine. The activity of CTP:phosphocholine cytidylyltransferase is also decreased in the hindlimb muscle of mutant mice. Concomitantly, the activities of PC phospholipase C and phospholipase A₂ are increased. The mitochondria in Chkb^−/− mice are abnormally large and exhibit decreased inner membrane potential. Despite the muscular dystrophy in Chkb^−/− mice, we observed increased expression of insulin like growth factor 1 and proliferating cell nuclear antigen. However, regeneration of hindlimb muscles of Chkb^−/− mice was impaired when challenged with cardiotoxin. Injection of CDP-choline increased PC content of hindlimb muscle and decreased creatine kinase activity in plasma of Chkb^−/− mice. We conclude that the hindlimb muscular dystrophy in Chkb^−/− mice is due to attenuated PC biosynthesis and enhanced catabolism of PC.