Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “HOMOGENATES”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 325 records · Page 18Linked to original sources
PubMed1983

Beta-oxidation of C-6-C-10 fatty acids in rat liver homogenates measured by selected ion monitoring: effects of cyanide and clofibrate.

The beta-oxidation of C-6-C-10-fatty acids/acyl CoA/acylcarnitines in whole rat liver homogenates was determined by specific and simultaneous measurements of the C-6-C-10-fatty acids, i.e. hexanoic, octanoic and decanoic acids, in hydrolysed homogenates in relation to time in assays incubated with the above-mentioned substrates. Measurements were performed by a combined gas chromatographic mass spectrometric technique, i.e. selected ion monitoring. The rate of beta-oxidation of the C-6-C-10-fatty acids/acyl CoA/acylcarnitines were registered as the consumption rate of the added substrate. The conversion to the C-2-shortened beta-oxidation products was illustrated simultaneously. The rate of beta-oxidation of C-6-C-10-fatty acids was several times higher in homogenates from clofibrate-treated rats than in control rats, both in the presence and absence of 2.0 mmol l-1 cyanide. Cyanide caused a minor but significant decrease in the beta-oxidation rate in both control and clofibrate-treated rats. No differences were found between the beta-oxidation of decanoic acid and decanoyl CoA in homogenates from clofibrate-treated rats, whereas the degree of beta-oxidation of DL-decanoylcarnitine was halved compared with decanoic acid and decanoyl CoA.

Animals↗
PubMed2006

Continuous homogeneous asymmetric transfer hydrogenation of ketones: lessons from kinetics.

Is polymer enlargement of homogeneous catalysts a tedious task? Is not batch operation with homogeneous catalysts the optimum performance point for homogeneous catalysis? Is kinetic modelling relevant to more than academic questions in homogeneous catalysis? Can all answers for a given system be answered satisfactory? In the authors' view, answers to these questions are no, no, yes, and depends. Polymer enlargement allowed the continuous operation of transfer hydrogenation in a chemical membrane reactor with total turnover numbers of up to 2.6 x 10(3) and a space-time yield of 0.58 kg L(-1) d(-1) with an enantiomeric ratio of 26.8 (enantiomeric excess 92.8 %) for a conversion level of 80 %. This was predicted from simulation conducted with a model from kinetic batch experiments adopted for continuous application. These simulations for the polymer-enlarged and the unmodified catalyst show that achieving comparable performance cannot be obtained by batch operation.

Alcohols↗
PubMed1992

The terminator, an apparatus for simultaneous homogenization of 96 small seeds individually.

An apparatus was developed to homogenize 96 individual seeds of tomato, pepper, cabbages, and other seeds of similar size in a 96-well tissue culture plate in 45 s. The apparatus, named the terminator, consists of two parts: a base plate that can hold the 96-well tissue culture plate, and a vibrating part that is attached to a variable transformer. The seeds are homogenized by the vibrating action of 96 small homogenizers that are attached to the vibrating part and that fit perfectly into the wells of the tissue culture plate. The seed homogenates made in this way can be used for electrophoretic analysis of genetic variations of seed proteins.

Brassica↗
PubMed2003

New homogeneous HDL-cholesterol assay without the influence of high TG sample using the selective detergent to lipoproteins.

Homogeneous HDL-cholesterol assays have been developed and used widely in routine analysis, but they have been reported to give inaccurate results in patients with hypertriglyceridemia. Recently, a new assay based on a new principle without the influence of triglycerides has also been developed and commercialized. We evaluated the basic performance of this new homogeneous HDL-cholesterol assay and compared it with the conventional polyethylene glycol/cyclodextrin-modified enzyme (PEGME) method using high-triglyceride (TG) samples (TG>8000 mg/L). For samples showing a discrepancy with the conventional method, other precipitation and ultracentrifugation (UC) methods were also used to confirm the values. This new homogeneous assay is based on the selective solubilizing effect of detergent on the different lipoproteins. First, non-HDL free cholesterol is consumed by enzyme and is cleared as a colorless reactant. Then. HDL-cholesterol is selectively solubilized by lipoprotein-specific detergent and reacted with the enzyme. As a result, the precision of this new homogeneous assay was good (CV<2%) over the wide range, and the measurement range was 0 to 2000 mg/L. This method correlated well with the PEGME method, which is a conventional method for normolipidemic samples (y=0.97x-3.1, r=0.994, n=424). It also correlated well with the UC method (y=0.99x+0.3, r=0.989, n=53). Fourteen high-TG samples showed different results from those obtained by the PEGME method. Among these samples, one contained abnormal lipoproteins (probably due to the influence of drug therapy) and gave a significantly different result from that obtained by the PEGME method. However, the values obtained by other methods (precipitation and ultracentrifugation) agreed well with those obtained by this new method. In conclusion, this method shows a good basic performance and is useful for high-TG samples without any interference. Therefore, it is considered to be very practical for a routine test.

Cholesterol, HDL↗
PubMed1995

Presystemic bradykinin metabolism in sheep and rat nasal homogenates.

Bradykinin (BK) and its fragments BK(1-8), BK(1-7), and BK(1-5) were incubated with sheep nasal homogenates to investigate the extent of peptide metabolism within the nasal mucosa. The products for both bradykinin and BK(1-8) degradation were found to be BK(1-7) and BK(1-5). BK(1-7) was metabolized to BK(1-5) alone. The patterns of degradation suggest that the Pro7-Phe8 bond of bradykinin was hydrolyzed first, then BK(1-7) was further hydrolyzed to form BK(1-5). The metabolism of bradykinin in rat nasal homogenates and plasma was also investigated. BK(1-5) was the only metabolite measurable in the rat nasal homogenates, likely due to the activity of an endopeptidase. The reduction in the bradykinin degradation rate resulting from the inhibition of angiotensin converting enzyme (ACE) or carboxypeptidase N indicates that these enzymes participate in mucosal bradykinin metabolism to some degree. In comparison, the products of bradykinin hydrolysis in rat plasma were found to be BK(1-8), BK(1-7), and BK(1-5). These results indicate that the enzyme populations or/and activities vary significantly between different species and between different tissues within the same species. Although significant aminopeptidase activities were detected in the sheep nasal homogenates, bradykinin was not affected by their presence, since the N-terminal sequence of bradykinin is not susceptible to hydrolysis by most aminopeptidases.

Animals↗
PubMed1975

Oxidation of lipids and membranes II: Effects of oxidants on cerebral white matter homogenates.

Bovine cerebral white matter homogenates were oxidized by various oxidizing solutions of equal molarity and subsequently extracted with water. Most of the oxidants tested (K-dichromate, FeCL3, H2O2, O2, and chloroperbenzoic, ascorbic, performic, and periodic acids) rendered the various myelin constituents less extractable than the constituents of unoxidized control homogenates. KMnO4, and to a lesser degree hemoglobin, rendered myelin constituents more extractable with water than those of the control homogenates. The findings indicate that most of the oxidants produced stabilization of the lamellar pattern, probably through cross-linking and polymerization. KMnO4 and hemoglobin caused labilization and breakdown of the membranous structure. Proof that stabilization of membranes is caused by some oxidants and that fragmentation occurs by the action of KMnO4 and hemoglobin was obtained by the light scattering technique and by electron microscopy of the oxidized homogenates. Evidence obtained indicated that formation of hydrophobic end groups during oxidation favors polymerization, while prevalence of hydrophilic groups is associated with fragmentation.

Animals↗
PubMed1989

Prevalence of nucleoside diphosphate kinase autophosphorylation in human colon carcinoma versus normal colon homogenates.

The G-regulatory proteins of adenylate cyclase, tubulin, and the ras oncogene protein product require the production of GTP from ATP in order to exert their effects within the cell. This implies that the activity of nucleoside diphosphate kinase (NDPK) plays a major role in the regulation of cellular events requiring GTP and that the level of activity of this enzyme is critical. This report presents a simple method for trapping a specific isozyme of NDPK in its high-energy phosphorylated form (NDPK approximately P) using EDTA and demonstrates that this NDPK approximately P is tenfold higher in malignant colon tumor tissue than in normal colon tissue. This autophosphorylation of the 21,000 and 24,000 Mr subunits of NDPK occurs rapidly at 0 degrees C, will use either [gamma-32P]ATP, [gamma-32P]GTP, or the corresponding 8-azidopurine photoprobes, is intramolecular, displays saturation effects, and is prevented from forming if GTP gamma S is added. Dephosphorylation in the homogenate occurs rapidly upon addition of Mg2+ or any nucleoside-5'-diphosphate. The subunits autophosphorylated in the homogenates are mostly in the soluble phase, and they comigrate with the subunits of pure NDPK from human erythrocytes. Cross-addition of normal and malignant homogenates does not decrease the level of autophosphorylation of NDPK, which indicates that the level of NDPK approximately P may be a quantitative measure of the level of this specific NDPK isozyme form. Assays for NDPK activity show correspondingly elevated levels in the malignant homogenates. Using western blot and photoaffinity labeling techniques, we distinguished the NDPK approximately P subunits from two closely migrating GTP-binding proteins. These were identified as the ras gene protein product and a 20,000 Mr protein, which comigrates identically with ADP-ribosylating factor (ARF). The ARF also comigrates in a tight band that is phosphorylated by [gamma 32P]ATP or [gamma-32P]GTP when Mg2+ is present.

Amino Acids↗
PubMed2002

Homogeneous preparation encoding (HoPE) in multislice imaging.

Fast magnetization preparation techniques acquire a series of echoes after a single magnetization preparation. If these echoes are acquired from different slices using a multislice technique the change in the preparation state of the echoes due to relaxation effects leads to different contrast modification for each slice. Encoding different preparation states along the phase-encoding direction of each slice instead of acquiring each slice in a different preparation state is introduced as a general concept to obtain images of identical contrast and point-spread function. This can be realized either by cycling the slice excitation order several times over the total number of repetitions or by moving the point of time at which the preparation is applied within each repetition. One possible application of this method is chemical shift selective fat saturation imaging. A homogeneous fat suppression across a multislice volume could be achieved using a FLASH sequence at a repetition time of TR = 145 ms, including a single fat saturation preparation. Conventional fat saturated spin-echo imaging at any TR can be accelerated significantly by reducing the number of applied preparations per repetition. A further application of the homogeneous preparation encoding (HoPE) method is described that encodes the spatial self-saturation of the multislice excitation order homogeneously in all slices. Only a reduced number of slices of the total volume are excited in each repetition and the slice excitation order is continuously moved along the imaging volume. This method is applied for time of flight (TOF) imaging. Using a TONE-like series of flip angles for the slice excitations of each repetition homogeneous TOF images can be obtained on the basis of a multislice acquisition.

Abdomen↗
PubMed2004

Effects of end-ring/shield configuration on homogeneity and signal-to-noise ratio in a birdcage-type coil loaded with a human head.

We modeled four different end-ring/shield configurations of a birdcage coil to examine their effects on field homogeneity and signal-to-noise ratio (SNR) at 64 MHz and 125 MHz. The configurations are defined as: 1) conventional: a conventional cylindrical shield; 2) surrounding shield: a shield with annular extensions to closely shield the end rings; 3) solid connection: a shield with annular extensions connected to the rungs; and 4) thin wire connection: a shield with thin wires connected to the rungs. At both frequencies, the coil with conventional end-ring/shield configuration produces the most homogeneous RF magnetic (B1) field when the coil is empty, but produces the least homogeneous B1 field when the coil is loaded with a human head. The surrounding shield configuration results in the most homogeneous B1 and highest SNR in the coil loaded with the human head at both frequencies, followed closely by the solid connection configuration.

Equipment Design↗
PubMed2000

Characterization of a selective protein kinase C substrate derived from the MARCKS phosphorylation site domain for use in brain tissue homogenates.

Protein kinase C (PKC) isozymes play crucial roles in neuronal signal transduction and can regulate transmitter release, ion channels, neural development, and plasticity. In vitro assays of PKC are frequently used to associate PKC activity with cellular function, and the availability of selective PKC substrates can facilitate such studies. We have characterized a commercially available 12 amino acid peptide derived from the myristoylated alanine-rich C kinase substrate (MARCKS-PSD, Calbiochem) for use in crude rat brain homogenates. Assays were performed at 25 degrees C for 10 min (linear up to 12 min) using optimal concentrations of calcium and lipid cofactors. Kinetic analysis of MARCKS-PSD phosphorylation by PKC purified from rat brain gave a K(m) of 2.3 microM, which was similar to the K(m) of 2.8 microM obtained using rat brain cortical homogenates. The selective PKC inhibitor bisindolylmaleimide reduced phosphorylation of MARCKS-PSD in a concentration-dependent manner, with greater than 95% inhibition at 1.0 microM. MARCKS-PSD was more potent than another widely used selective PKC substrate (neurogranin((28-43)) and was a good substrate for human recombinant PKC alpha, delta, and epsilon but not zeta. The ontogeny of PKC activity was examined in the cortex and cerebellum. PKC activity was low at birth and reached adult levels by 21 days of age in both regions. Calcium-independent PKC activity in brain homogenates could be measured with MARCKS-PSD and accounted for approximately 25 and 10% of total activity in 1-day-old and adult rat cortex, respectively. These results suggest that the MARCKS-PSD peptide can be used as a selective PKC substrate in rat brain homogenates.

Animals↗
PubMed2001

Green fluorescent protein mutant as label in homogeneous assays for biomolecules.

The green fluorescent protein (GFP) and its mutants have been extensively used to study various cellular processes and, more recently, as labels in binding assays. We have employed a mutant of GFP, an enhanced GFP (EGFP), in the development of homogeneous assays for biotin and cortisol. To demonstrate the feasibility of using EGFP as a label with different kinds of binders in the development of homogeneous assays, we employed the biotin-avidin and an antigen-antibody as the binding pairs. Biotin and cortisol were chemically conjugated to EGFP. A quenching of fluorescence intensity of EGFP was observed upon binding of avidin to the EGFP-biotin conjugate. The percentage fluorescence quenching observed decreased as the concentration of free biotin in the sample increased due to the fewer binding sites on avidin available for binding to the EGFP-biotin conjugate. A dose-response curve for biotin was generated by relating percentage fluorescence quenched with free biotin in the sample. To determine whether EGFP can undergo a similar type of homogeneous change when used as a label for antigen-antibody type of binding, cortisol was selected as a model analyte. In the presence of an anti-cortisol antibody the fluorescence signal of the EGFP-cortisol conjugate was quenched. A dose-response curve for cortisol was generated by relating the quenching in the fluorescence signal with varying amounts of free cortisol in the sample. This is the first time that GFP or one of its mutants has been employed as a label in homogeneous assays, thus enhancing the versatility of employing GFP or its mutants in a number of bioanalytical applications, such as clinical analysis and high-throughput screening systems.

Biotin↗
PubMed2001

Pellet pestle homogenization of agarose gel slices at 45 degrees C for deoxyribonucleic acid extraction.

A simple method for extracting DNA from agarose gel slices is described. The extraction is rapid and does not involve harsh chemicals or sophisticated equipment. The method involves homogenization of the excised gel slice (in Tris-EDTA buffer), containing the DNA fragment of interest, at 45 degrees C in a microcentrifuge tube with a Kontes pellet pestle for 1 min. The "homogenate" is then centrifuged for 30 s and the supernatant is saved. The "homogenized" agarose is extracted one more time and the supernatant obtained is combined with the previous supernatant. The DNA extracted using this method lent itself to restriction enzyme analysis, ligation, transformation, and expression of functional protein in bacteria. This method was found to be applicable with 0.8, 1.0, and 2.0% agarose gels. DNA fragments varying from 23 to 0.4 kb were extracted using this procedure and a yield ranging from 40 to 90% was obtained. The yield was higher for fragments 2.0 kb and higher (70-90%). This range of efficiency was maintained when the starting material was kept between 10 and 300 ng. The heat step was found to be critical since homogenization at room temperature failed to yield any DNA. Extracting DNA with our method elicited an increased yield (up to twofold) compared with that extracted with a commercial kit. Also, the number of transformants obtained using the DNA extracted with our method was at least twice that obtained using the DNA extracted with the commercial kit.

DNA↗
PubMed2000

A Model for Simultaneous Homogeneous and Heterogeneous Nucleation in the Case of Slow Reaction Rate.

Our previous model for simultaneous homogeneous and heterogeneous nucleation, which was successfully tested for silver particle generation by a chemical reduction method, is extended to the case of slow reaction rate. Nucleation in a system containing seed particles in the case where the reaction rate is slow occurs according to the following rule: homogeneous nucleation is predominant when n(p)/n(0)(*),>1 (n(p) is the seed number concentration, and n(0)(*) the number concentration of homogeneously nucleated particles without seed particles), and heterogeneous nucleation is predominant when n(p)/n(0)(*)<1. This theoretical prediction is in good agreement with the experimental results obtained for zinc sulfide particles generated by homogeneous precipitation. Copyright 2000 Academic Press.

Journal Article↗
PubMed1996

Ryanodine perfusion decreases cardiac mechanical function without affecting homogenate sarcoplasmic reticulum Ca2+ uptake: comparison with the stunned heart.

This study tested the hypothesis that perfusion with low concentrations of ryanodine, which opens the sarcoplasmic reticulum (SR) Ca2+ channel in a sub-conducting state, could mimic the effects of stunning on both mechanical and SR activity. Perfusion of isolated rat hearts with 10-160 nM ryanodine progressively decreased left ventricular developed pressure (LVDP) and increased end-diastolic pressure (EDP), but LVDP decreased more and EDP increased less than in the stunned heart. The effect of ryanodine perfusion on LVDP and EDP is consistent with the opening of the SR Ca2+ channel by high-affinity ryanodine binding, reducing SR Ca2+ content and interfering with mechanical function. In contrast to stunning, ryanodine perfusion did not affect the homogenate Ca2+ uptake rates measured in the presence or absence of high [ryanodine]. Perfusion with 80 nM 3H-ryanodine resulted in a large decline in LVDP, but only a small degree of ryanodine binding. Thus, prolonged opening of only a few channels affects the SR in situ, whereas this is undetectable in the homogenate. Higher levels of ryanodine binding (0.3 pmol/mg) to the in vitro homogenate also did not affect the homogenate Ca(2+)-uptake rate in the presence or absence of high [ryanodine], whereas it reduced the stimulation of Ca2+ uptake by ruthenium red. High-affinity ryanodine binding to the SR Ca2+ channel, either by perfusion or by binding after homogenisation, did not duplicate the increased Ca2+ efflux observed in the stunned heart, suggesting that the SR defect in the stunned heart is not a prolonged opening of a sub-conducting state of the SR Ca2+ channel.

Animals↗
PubMed1995

Differential homogenization and amplification of two satellite DNAs in the genus Cucurbita (Cucurbitaceae).

Two different satellite DNAs exist in the genus Cucurbita which are different with respect to repeat length (350 bp and 170 bp), array size, and sequence homogenization. Whereas the 350-bp satellite DNA is prominent and very homogeneous in all species investigated except for C. maxima and C. lundelliana, the 170-bp satellite is rather evenly distributed in all species. In C. maxima and C. lundelliana the 350-bp satellite is present only in small amounts, but detectable by the sensitive PCR method. These repeats are also very homogeneous, reflecting a silent stage of satellite DNA. In contrast, the 170-bp satellite DNA is intra- and interspecifically heterogeneous. It is striking that the species with no detectable amount of 350-bp satellite contain 170-bp satellite DNA clusters with the highest degree of homogeneity. The evolution of satellite DNA repeats within cultivated and wild species in the genus Cucurbita is elucidated using the sequence data of both satellite DNAs from all species investigated. The value of satellite DNA for phylogenetic analysis between closely related species is discussed.

Base Sequence↗
PubMed1977

Adenosine triphosphate catabolism in homogenates of rat secretory enamel organs incubated in histochemical lead media.

To investigate how lead, when used as trapping agent, influences the ATP hydrolysis and to study how ATP is catalyzed in histochemical systems, homogenized secretory enamel organs were incubated in histochemical [3H]-ATP media. Aliquots from the media were taken after 3, 10, 20 and 30 min, the ATP and formed metabolites were separated by electrophoresis and radiometrically quantitated. In media lacking both lead and homogenate 2% of the ATP was spontaneously hydrolyzed during 30 min incubation at room temperature. The presence of lead caused an additional 8% hydrolysis at pH 7.2 and an additional 20% hydrolysis at pH 9.4. In the presence of homogenate, however, lead caused a net decrease of the hydrolysis of ATP as well as of ADP and AMP. This enzyme inhibition varied from around zero to some 80%, depending on pH and substrated involved. In homogenate-containing lead media, at both pH 7.2 AND 9.4, ATP was rapidly hydrolyzed primarily to ADP and subsequently to AMP and adenosine and/or inosine. After 5--10 min ADP constituted the predominant substrate at both pH:s. At pH 7.2 ADP remained so for the rest of the incubation, whereas at pH 9.4 AMP was predominant substrate at the end of the incubation. AMP was the finan catabolic product in experiments at pH 7.2, and adenosine and/or inosine at pH 9.4. Inorganic phosphate was liberated almost linearly during the whole incubation period. The results indicate that histochemical studies of substrate specific ATP-ases should be performed with short incubation times and, when high specific activities are present, in large quantities of incubation media to reduce interference by ADP and AMP hydrolyzing enzymes.

Adenosine Diphosphate↗
PubMed1985

Theoretical consideration of drug distribution kinetics in a noneliminating organ: comparison between a "homogeneous (well-stirred)" model and "nonhomogeneous (tube)" model.

Drug distribution kinetics in a noneliminating organ or tissue has been mathematically examined. The homogeneous (well-stirred) model regards the noneliminating organ or tissue as a homogeneous compartment in which the drug is equilibrated with that in the blood leaving the organ or tissue. The nonhomogeneous (tube) model views the noneliminating organ or tissue as comprising a number of parallel cylindrical tubes containing binding sites distributed homogeneously along these tubes. These two models are examined, considering the pseudo-distribution equilibrium phase after bolus injection and a linear binding condition. Although both models predict a similar tissue distribution under a variety of conditions, significant differences exist in the predictions of various pharmacokinetic parameters as a function of the drug distribution, such as blood flow, organ volume, slope of the terminal phase, and the tissue-to-blood partition coefficients. The predictability and limitations of these two models are explored. Distribution characteristics of the two models are also examined for adriamycin, actinomycin D, tetrachlorobiphenyl, hexachlorobiphenyl, digoxin, and ethoxybenzamide; no difference is observed. It is concluded that the assumption of a homogeneous (well-stirred) compartment is suitable for describing the drug distribution kinetics of these drugs.

Kinetics↗
PubMed1997

Biological activation of N(G)-nitro-D-arginine by kidney homogenate .

N(G)-nitro-L-arginine (L-NNA) and D-NNA have been shown to inhibit endothelium-dependent relaxation. This study examined if the inhibitory effect of L-NNA or D-NNA on relaxation is increased following incubation of the drug with the supernatant of tissue homogenates. Acetylcholine (ACh) caused concentration-dependent relaxation of pre-constricted rat aortic rings with maximum relaxation of 95%. Maximum relaxations to ACh were reduced to 71 and 37% in the presence of D-NNA (40 microM) and L-NNA (1 microM), respectively. Relaxation to ACh was further reduced to 18% in the presence of D-NNA that was incubated for 1 h with the supernatant of kidney homogenate, but unaffected by D-NNA incubated with the supernatant of trichloroacetic acid-denatured kidney homogenate. Incubation of L-NNA (1 microM) with either kidney supernatant or denatured kidney supernatant for 1 h did not affect its inhibitory effect on ACh-induced relaxation. Neither 1 h's incubation with plasma, or supernatants of liver, lungs or aorta homogenates affected the inhibitory action of D-NNA (40 or 120 microM) on ACh-induced relaxation. After D-NNA was incubated in kidney supernatant, its inhibitory effect on ACh-induced relaxation of the aorta was abolished by pretreatment of the aorta with L-arginine (L-Arg) but not D-Arg suggesting involvement of the L-Arg pathway. The results suggest that D-NNA is converted by the kidney to a compound that acts similar to L-NNA. There appears to be little conversion of L-NNA to D-NNA.

Acetylcholine↗