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Compositional changes of fatty acids in the 1(1")-and 2(2")-positions of cardiolipin from liver, heart, and kidney mitochondria of rats fed a low-fat diet.

Cardiolipins from liver, heart and kidney mitochondria of rats fed a fat-free diet for 66 days have been analyzed for their fatty acid composition and positional distribution. The main effect was a dramatic decrease of linoleic acid which was counterbalanced by increases in the levels of palmitoleic, oleic and cis-vaccenic acids. Linoleic acid remains asymmetrically distributed between positions 1(1") and 2(2") with a positive selectivity for positions 1(1"). Its decrease is considerably faster in positions 2(2") than in positions 1(1"), which would suggest different rates of fatty acid turnover. Fat deficiency induces the appearance of 18:2(n-7) and a significant increase of 20:3(n-6) (dihomo-gamma-linolenic acid) in liver and kidney cardiolipins. In contrast, 20:3(n-6) level remains unchanged in other mitochondrial phospholipids. 18:2(n-7) and 20:3(n-6) are almost evenly distributed between both pairs of positions. Both acids have a common structural feature, that is double bonds in positions 8 and 11. 20:3(n-9) accumulates in large amounts in other mitochondrial phospholipids, but not in cardiolipins. Although surprising, 20:3(n-6) has thus to be considered as a specific marker of deficiency for cardiolipins when it is esterified to positions 1(1"). Taking into account various analytical data, it would appear that positions 1(1") of cardiolipins can only incorporate unsaturated fatty acids containing at least one cis double bond in position 8 or 9, with no other double bond between these positions and the carboxylic group.

Animals↗

Occurrence of a furan fatty acid in marine bacteria.

A fatty acid containing a furan ring was detected in the cellular lipids of marine bacteria, Shewanella putrefaciens, Marinomonas comunis, Enterobacter agglomerans, Pseudomonas fluorescens, etc., which were isolated from the intestinal liquor of fishes. Analytical data indicated that the fatty acid was 10,13-epoxy-11-methyloctadeca-10, 12-dienoic acid. Therefore, we propose that furan fatty acids detected in marine fish are derived not only from marine plants but also from intestinal bacteria of fishes.

Animals↗

Effects of phenytoin on the in vivo kinetics of thiamine and its phosphoesters in rat nervous tissues.

The in vivo effects of chronic (30 days) and subchronic (10 days) intragastric treatment with phenytoin (PHT) (500 mg/kg) b.wt., suspended in 10% arabic gum water solution) on the uptake and metabolism of thiamine (T), T monophosphate (TMP) and T pyrophosphate (TPP) were evaluated in rat nervous regions (cerebral cortex, brainstem, cerebellum and sciatic nerve) by determining the radioactivity of T and its phosphoesters in plasma and tissues at fixed time intervals (0.25-240 h) after an i.p. injection of thiazole-[2-14C]thiamine (30 micrograms: 1.25 microCi). A nutritionally adequate diet containing T in excess was given to the animals in order to produce a virtually stable content of T compounds in the tissues. Analytical data were processed by using a compartmental model which allowed the calculation of fractional rate constants (FRC), turnover rates (TR) and turnover times. Compared with vehicle-treated controls, animals treated chronically with PHT exhibited lower levels of radiolabelled T compounds in all nervous regions except for the cerebral cortex. These alterations were not found in animals receiving subchronic treatment. Evaluation of FRC values indicated that PHT-induced effects on T metabolism differed depending on the length of PHT treatment and the nervous region considered. Overall, PHT appeared to interfere mainly with T and TMP uptake, TPP dephosphorylation to TMP and TPP turnover times, these effects being particularly prominent in the cerebellum and in the brainstem of chronically treated animals. Since all changes in T uptake and metabolism were observed in the absence of overt behavioural toxicity, these findings may have potential clinical relevance in highlighting possible mechanisms by which PHT therapy can alter brain metabolism.

Animals↗

Polysaccharides from Peptostreptococcus anaerobius and structure of the species-specific antigen.

The cell-envelope antigens of Peptostreptococcus anaerobius were extracted from intact cells by autoclave or alkaline treatment. The purified species-specific antigen (G) was identified among several polysaccharides obtained from the extracts by successive treatments with ribonuclease and pronase followed by ion-exchange and gel-filtration chromatography. G was investigated by 13C- and 31P-n.m.r. spectroscopy, titrimetry, elemental analysis, and gas-liquid chromatography. Oxidation of G with NaIO4 followed by reduction with NaBH4 and mild acid hydrolysis yielded the Smith degradation product of G (GS). Treatment of G and GS with 48% HF gave the respective dephosphorylated products GF and GSF. The structures of GS, GF, and GSF were investigated by 13C-n.m.r. spectroscopy, methylation analysis, and gas-liquid chromatography-mass spectrometry. The principal constituents of G were 2-acetamido-2-deoxy-D-glucose (D-GlcNAc), D-glyceric acid, and phosphate as a diester, in the ratio 2:1:1, and a minor amount of D-glucose (beta-D-Glcp). GS contained D-GlcNAc, D-glyceric acid, glycerol, and phosphate in a 1:1:1:1 ratio. GF and GSF contained D-GlcNAc and D-glyceric acid in the ratios 2:1 and 1:1, respectively. A structure for the principal repeating unit of polymeric G compatible with the analytical data consists of alpha-D-GlcpNAc-(1----3)-alpha-D-GlcpNAc-(1----2)-D-glyceric acid units linked through C-6'-C-6" phosphate diester bridges. This structure is novel for two reasons: (a) unsubstituted glyceric acid residues occur as aglycons in the repeating structure, and (b) phosphate diester bridges link nonanomeric glycose carbons in a non-nucleic acid polymer. The structural role of the minor amount of beta-D-Glcp in G remains unknown.

Antigens, Bacterial↗

Measurement by radioimmunoassay of unconjugated estriol in pregnancy serum.

Antisera to estriol 6--onjugates were tested for suitability in the direct radioimmunoassay of unconjugated estriol in extracts of pregnancy serum. Assessment of specificity through titration with competing steroids allowed a selection to be made within the group of antisera. The measurement was then standardized by checking the absence of analytical blank (solvent and sample blank) and the extent of binding variability associated with the use of charcoal-dextran as a separating agent. Validation was made by means of the usual recovery and dilution tests, and by a cross-comparison of analytical data obtained with different antisera; reproducibility of calibration curve and within-and between-assay variability was evaluated under routine conditions. The validity of the clinical information was assessed by assaying 202 samples randomly collected throughout normal pregnancy from the 16th week to term: both trend and levels of unconjugated estriol concentration were found to be in good agreement with the literature data.

Animals↗

The use of the simplex method for the optimisation of non-linear functions on a laboratory microcomputer.

The use of the microcomputer for the processing of analytical data in the laboratory is currently increasing. However, computational methods which have been used successfully on minicomputers may not be suitable for microcomputer implementation due to the lack of available precision. This paper describes a technique, namely the simplex method, which may be successfully applied to the processing of laboratory data on an 8-bit microcomputer. The method is used to optimise a non-linear function in order to fit a curve to radioimmunoassay (RIA) and immunoradiometricassay (IRMA) data. The microcomputer package described has been tested on laboratory data and is currently in use in a clinical biochemistry laboratory.

Biometry↗

The CuAsite of the caa3-type oxidase of Bacillus subtilis is a mixed-valence binuclear copper centre.

A copper-containing domain of the caa3-type oxidase from Bacillus subtilis has been expressed as a water-soluble protein in the cytoplasm of Escherichia coli. Electron paramagnetic resonance (EPR) spectra of this purple domain show well-resolved lines in the gz resonance, both at X-band and S-band frequencies. Interpretation of EPR spectra and analytical data indicate a binuclear copper site consisting of one Cu2+ and one Cu1+. This copper site closely resembles CuA in subunit II of cytochrome c oxidase and is shown here to be a mixed-valence [Cu2+-Cu1+] binuclear centre.

Bacillus subtilis↗

Photoionization assessment of C3-C5 alkadienes and alkenes in urban air.

Hydrocarbons from samples of traffic-polluted urban air were separated by gas chromatography on an aluminium oxide column and assessed simultaneously by photoionization detection (PID) and flame ionization detection (FID) after effluent splitting. The 10.2 eV photoionization detector selectively detects alkadienes and alkenes but not alkanes and alkynes in the C3-C5 region. The maximum PID/FID response ratio for alkadienes and alkenes is also obtained in this region. The analytical system as a whole is particularly favourable for the C3-C5 alkenes. Analytical data are given for propadiene, 1,3-butadiene, propene, butenes and pentenes.

Air Pollutants↗

Purification and analytical characterization of an anti-CD4 monoclonal antibody for human therapy.

A purification process for the monoclonal anti-CD4 antibody MAX.16H5 was developed on an analytical scale using (NH4)2SO4 precipitation, anion-exchange chromatography on MonoQ or Q-Sepharose, hydrophobic interaction chromatography on phenyl-Sepharose and gel filtration chromatography on Superdex 200. The purification schedule was scaled up and gram amounts of MAX.16H5 were produced on corresponding BioPilot columns. Studies of the identity, purity and possible contamination by a broad range of methods showed that the product was highly purified and free from contaminants such as mouse DNA, viruses, pyrogens and irritants. Overall, the analytical data confirm that the monoclonal antibody MAX.16H5 prepared by this protocol is suitable for human therapy.

Animals↗

Nucleotide sequences of immunoglobulin-epsilon pseudogenes in man and apes and their phylogenetic relationships.

To understand the phylogenetic relationships between hominoids, the nucleotide sequences of immunoglobulin-epsilon processed pseudogenes from chimpanzee, gorilla and orangutan were determined. The basic structures of these processed pseudogenes agreed with their human counterpart. Although the degrees of nucleotide differences between man and the African apes had no statistical significance, all the analytical data examined supported the theory that chimpanzee is the closest relative of man. This result was consistent with that deduced by our recent qualitative study. Studies on the nucleotide sequences of globin genes have suggested that the molecular clock runs more slowly in hominoids than in non-hominoid primates. According to the present data, however, further retardation of the evolutionary rate was not observed in the human lineage. Assuming that orangutan diverged 14 million years ago and that the evolutionary rate between the orangutan lineage and the lineage leading to the other three species is constant, the divergence dates of chimpanzee and gorilla were estimated to be 4.9(+/- 0.9) and 5.9(+/- 0.9) million years ago, respectively.

Animals↗

Toward a biochemical classification of depressive disorders--VIII. Platelet monoamine oxidase activity in subtypes of depressions.

Platelet monoamine oxidase (MAO) activity was examined in 77 depressed patients (40 males and 37 females) and 28 controls (14 males and 14 females). Patients were compared across increasingly specific diagnostic groupings in a four-step data analytic procedure. In step 1, MAO activity in the total sample of depressed patients was compared with that of control subjects. In step 2, Unipolar depressed patients were compared with Bipolar (Bipolar I and Bipolar II) depressed patients. In step 3, Unipolar depressed patients with and without schizotypal features were compared. In step 4, both the nonschizotypal Unipolar patients and compared. In step 4, both the nonschizotypal Unipolar patients and nonschizotypal Bipolar patients were separated into those who met RDC criteria for a definite Endogenous depression and those who did not; and platelet MAO activity was compared in the resulting four groups. Results indicated significantly higher platelet MAO activity in nonschizotypal Unipolar Endogenous depressed patients than in nonschizotypal Bipolar Endogenous depressed patients or nonschizotypal Unipolar Other patients. In addition, the presence of a definite Endogenous depressive syndrome was associated with greater overall symptom severity in both Unipolar and Bipolar depressed patients. Findings are discussed with respect to the conflicting results reported in previous studies of MAO activity in patients with depressive disorders.

Adolescent↗

Interacting neurotransmitter systems. A non-experimental approach to the 5HIAA-HVA correlation in human CSF.

The repeatedly observed strong positive correlation between 5-hydroxyindoleacetic acid (5HIAA) and homovanillic acid (HVA) in human cerebrospinal fluid (CSF) prompted an investigation to see if conclusions concerning possible interactions between brain serotonin and dopamine turnover could be reached from human CSF concentrations of these acid metabolites. CSF data from patients with depressive disorders diagnosed according to the RDC from Sweden (n = 140) and from the National Institute of Mental Health (n = 35) were used to test structural hypotheses by two statistical approaches--LISREL analysis and logistic regression. Results from both men and women were unequivocal: 5HIAA "controls" HVA, interpretable as a regulatory action of serotonin turnover on dopamine turnover. In women, only 5HIAA was affected by age, height and body size (higher in elderly, short and stout women); no similar relationships were seen in males. The concept of a serotonergic regulation of dopamine turnover was tested on brain punch analyses of serotonin and dopamine and their metabolites in two sets of dogs in a large number of brain areas. Results confirm a facilatory effect of serotonin on indices of dopamine turnover in many brain regions, especially brain stem and hypothalamus. The animal data validate the data analytic approach in humans.

Adolescent↗

Quantitative analytical mass spectrometry of endogenous neuropeptides in human pituitaries.

This manuscript reviews state-of-the-art mass spectrometric (MS) methodology for the qualitative (amino acid sequence determination) and quantitative analysis of opioid neuropeptides in human pituitary tissue. Those analytical data are required for the elucidation of the basic molecular mechanisms involved in tumor formation and to test the hypothesis that metabolic defects in neuropeptidergic system processing is a contributing factor to human anterior pituitary tumor formation. Several different neuropeptide products that derive metabolically from the proenkephalin A and proopiomelanocortin (POMC) precursors have been analyzed separately and together in human pituitaries, including post-mortem controls and post-surgical tumors. The quantification, with optimal molecular specificity, of a peptide in a tissue is an important measurement because the amount of an endogenous peptide reflects the ratio of its synthesis to its degradation and thus, any defects in those processes may be reflected in the amount of a peptide.

Amino Acid Sequence↗

Dimerisation of levonorgestrel in solid state ultraviolet light irradiation.

UV-irradiation of levonorgestrel (1) in the crystalline state under a nitrogen atmosphere yielded its dimer, [17 alpha[1R-(1 alpha,2 beta,4a beta,4b alpha,10a alpha)]]-13-ethyl-17- [4- (2-ethyl-1,2,3,4a,4b,5,6,7,9,10,10a-dodecahydro-7-oxo-1-phenant renyl)-1- methylen-2-oxo-butoxy]-18,19-dinorpregna-4-en-20-in-3-one(3) , as the principal photoproduct. It was characterized from its spectral and analytical data. The single crystal X-ray crystallographic data of 1 indicated the possibility of its photochemical dimerisation.

Crystallography, X-Ray↗

Two-eyed versus one-eyed salamanders: does binocularity enhance the optically evoked skin blanching reactions of Ambystoma larvae?

A wide variety of visual functions show increases attributable to binocularity, and the question pursued here was whether a second eye enhances the visually stimulated skin blanching reaction of the larval salamander. Dermal melanin spots (produced by the aggregations of melanosomes within dermal melanophores and which contract or expand to lighten or darken the skin) were measured in eyeless (controls), one-eyed and two-eyed Ambystoma punctatum larvae after chronic adaptation of the subjects to a white background (i.e., stimulus conditions for maximum blanching). The eyeless subjects showed no blanching (thus remained dark) in white cups, and they exhibited melanin spots 7 or 8 times the size of those of the other two groups. All one-eyed or two-eyed subjects exhibited blanching reactions; planometric comparison revealed a significantly larger melanin spot area for one-eyed than for two-eyed animals; i.e., the binocular condition permitted greater contraction of the pigment spots than did the monocular condition. Analytical data compared favorably with independently ascertained pigmentation indices. The results indicate that a second eye quantitatively elevates the blanching maximum of a larval salamander.

Ambystoma↗

Composition of the gum from Combretum paniculatum and four other gums which are not permitted food additives.

Only three gum exudates are permitted for pharmaceutical and food use by international regulatory authorities, viz. gum tragacanth (Asiatic Astragalus spp.), gum karaya (Sterculia spp.) and gum arabic [Acacia senegal (L.) Willd.], but a wide range of other tree exudates is used for a variety of uses in their countries of origin. This paper presents analytical data for the gum exudates from Atalaya hemiglauca, Cassine aethiopica, Combretum paniculatum, Sclerocarya birrea, and Pseudocedrela kotschyi. These gums may have local technological applications, but are not recommended for addition to foodstuffs.

Amino Acids↗

Lisianthus flavonoid pigments and factors influencing their expression in flower colour.

NMR, MS and analytical data are cited in support of the newly defined complete structures of the major flavonoid pigments and copigments in lisianthus flowers. The copigments newly characterized and found in flowers of all colours are kaempferol-3-O-beta-D-[6-O-rhamnopyranosyl-4-O-E-p- coumaroylgalactopyranoside]-7-O-alpha-L-rhamnopyranoside, its Z-isomer and by analogy, the lesser isorhamnetin and myricetin equivalents. Purple flower pigments with newly defined structures are: delphinidin-3-O-beta-D-[6-O-alpha- L-rhamnopyranosylgalactopyranoside] 5-O-beta-D-[6-E-p- coumaroylglucopyranoside], its Z-isomer, and by analogy the lesser cyanidin equivalent, together with delphinidin-3-O-beta-D-galactopyranoside-5-O-beta-D-[6-E-p- coumaroylglucopyranoside], its Z-isomer, and by analogy the lesser cyanidin equivalent. Different pigment/copigment compositions are shown to account for the basic colour differences between white, cream, pink, mauve and purple flowers, but other factors involved in stabilizing and fine-tuning the colours are pigment concentration, copigmentation and pH control.

Anthocyanins↗

Nucleoside conjugates. 8. The preparation of 5-fluoro-2'-deoxyuridine conjugates of corticosteroids.

Three 5'-(steroid-21-phosphoryl)-5-fluoro-2'-deoxyuridines (VI-VIII) have been prepared and characterized by uv, ir, 1H-nmr, elemental analysis, chemical and enzymatic hydrolyses. These new compounds are 5-fluoro-2'-deoxyuridine conjugates of cortisol (VI), cortico-sterone (VII), and prednisolone (VIII). Besides the physical and analytical data, all of the conjugates were demonstrated to be enzymatically hydrolyzed to the corresponding steroid and 5-fluoro-2'-deoxyuridine 5'-monophosphate (III), and the latter was further shown to be hydrolyzed to 5-fluoro-2'-deoxyuridine (II) by phosphodiesterase I, 5'-nucleotidase, and acid phosphatase. However, they were shown to be resistant to hydrolysis by bacterial alkaline phosphatase.

Antineoplastic Agents↗