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Letteratura scientifica selezionata sul tema "Isoenzymes Analysis"

Autore: Grafiati
Pubblicato: 4 giugno 2021
Ultima modifica: 28 gennaio 2023

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Articoli di riviste sul tema "Isoenzymes Analysis"

1

Tillyer, C. R., S. Rakhorst e C. M. Colley. "Multicomponent analysis for alkaline phosphatase isoenzyme determination by multiple linear regression". Clinical Chemistry 40, n. 5 (1 maggio 1994): 803–10. http://dx.doi.org/10.1093/clinchem/40.5.803.

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Abstract Alkaline phosphatase (EC 3.1.3.1) isoenzymes in serum may be determined by multicomponent analysis of the enzyme activities in the presence of multiple inhibitors. To determine inhibition coefficients of the isoenzymes, we used multiple linear regression analysis to compare alkaline phosphatase activities in the presence of known inhibitors with electrophoretically determined isoenzyme activities in plasma and serum samples. All possible combinations of exactly determined and overdetermined linear systems of inhibitors were ranked according to their prediction error to select an optimum set. The best multicomponent system for prediction included the use of levamisole, phenylalanine, and heat inhibition at 56 degrees C and 65 degrees C to determine bone, hepatic, intestinal, and placental isoenzymes. Consideration of the hepatic isoenzyme as liver and macromolecular fractions resulted in significantly worse predictions. Error analysis involving repeat determinations and a simplex optimization of the inhibition coefficients indicated that the inaccuracy of the comparison electrophoretic method may have been a major factor affecting poor isoenzyme prediction in some samples.
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2

Dunaway, G. A., T. P. Kasten, T. Sebo e R. Trapp. "Analysis of the phosphofructokinase subunits and isoenzymes in human tissues". Biochemical Journal 251, n. 3 (1 maggio 1988): 677–83. http://dx.doi.org/10.1042/bj2510677.

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The 6-phosphofructo-1-kinase (PFK) subunits and isoenzymes were studied in human muscle, heart, brain, liver, platelets, fibroblasts, erythrocytes, placenta and umbilical cord. In each tissue, the subunit types in the native isoenzymes were characterized by immunological titration with subunit-specific antibodies and by column chromatography on QAE (quaternary aminoethyl)-Sephadex. Further, the subunits of the partially purified native isoenzymes were resolved by SDS/polyacrylamide-gel electrophoresis, identified by immunoblotting, and quantified by scanning gel densitometry of silver-stained gels and immunoblots. Depending on the type of tissue, one to three subunits were detected. The Mr values of the L, M and C subunits regardless of tissue were 76,700 +/- 1400, 82,500 +/- 1640 and 86,500 +/- 1620. Of the tissues studied, only the muscle PFK isoenzymes exhibited one subunit, which was the M-type subunit. Of the other tissues studied, the PFK isoenzymes contained various amounts of all three subunits. Considering the properties of the native PFK isoenzymes, it is clear that, in human tissues, they are not simply various combinations of two or three homotetrameric isoenzymes, but complex mixtures of homotetramers and heterotetramers. The kinetic/regulatory properties of the various isoenzyme pools were found to be dependent on subunit composition.
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Yoshikuni, Keiko, Takashi Matsuda, Janka Poracova, Akemi Sakai, Keiko Shimada, Noriko Tabuchi e Kiyoh Tanishima. "Phylogenic Study of Denaturation of Lactate Dehydrogenase Isoenzymes from Different Species by High and Low Temperature". Annals of Clinical Biochemistry: International Journal of Laboratory Medicine 38, n. 5 (settembre 2001): 548–53. http://dx.doi.org/10.1177/000456320103800513.

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We studied the influence of storage at different temperatures on lactate dehydrogenase (EC 1.1.1.27; LD) isoenzymes from different tissues and different species, and analysed biochemical and biophysical mechanisms of denaturation during storage. Isoenzymes obtained from tissue extracts of mammals, poultry, reptiles, amphibians and fish were shown to have their own denaturation ranges at low temperatures by post-treatment assays and transition temperature analysis. These ranges were between − 10 and − 20°C for most vertebrate LD isoenzymes. Circular dichroism analysis indicated that the denaturation of LD isoenzymes was probably caused by a change in the hydrophobic interactions in the molecule. At higher temperatures, LD-1 isoenzyme was more thermostable than LD-5 from the same animal species, except for rats, the LD-5 activity of which was more thermostable than the LD-1 activity. These findings indicate that variable effects of storage of samples and reference materials at low temperatures should be considered, and that it is necessary to establish LD isoenzyme standards for animal clinical laboratory investigations.
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4

Giannoulaki, E. E., D. L. Kalpaxis, C. Tentas e P. Fessas. "Lactate dehydrogenase isoenzyme pattern in sera of patients with malignant diseases." Clinical Chemistry 35, n. 3 (1 marzo 1989): 396–99. http://dx.doi.org/10.1093/clinchem/35.3.396.

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Abstract Total lactate dehydrogenase (LDH; EC 1.1.1.27) activity and the percentage distribution of LDH isoenzymes were determined in 127 patients with malignant diseases. A shift in the isoenzyme patterns was observed toward the M-type, with an increase in the percentage of LDH-4 and LDH-5 isoenzymes and a slight increase in total LDH activity of all patients. Serum samples from 68 of the patients contained an abnormal isoenzyme of LDH, "LDH-1 ex," that, on agarose gel electrophoresis at pH 8.6, migrated between albumin and LDH-1 isoenzyme. Chemotherapy, radiotherapy, or surgical removal of the tumor was accompanied by disappearance of this abnormal isoenzyme. The heat stability of LDH-1 ex isoenzyme appears to be similar to that of LDH-1 but greater than that of the other LDH isoenzymes. Statistical analysis of these data demonstrated a significant correlation between malignancy and the appearance of LDH-1 ex isoenzyme (P less than 0.001). In contrast, the relationship between LDH-1 ex isoenzyme and metastasis or anatomical location of the malignancy is not statistically important (P less than 0.1).
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Bar, Jair, Stuart Spencer, Shethah Morgan, Laura Pike, David Cunningham, Jane D. Robertson, Juliane M. Jürgensmeier e Glenwood D. Goss. "Correlation of lactate dehydrogenase (LDH) isoenzyme profile with outcome in advanced colorectal cancer (CRC) patients (pts) treated with chemotherapy and bevacizumab (BEV) or cediranib (CED)." Journal of Clinical Oncology 30, n. 15_suppl (20 maggio 2012): e13541-e13541. http://dx.doi.org/10.1200/jco.2012.30.15_suppl.e13541.

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e13541 Background: There is a clinical need for predictive biomarkers of efficacy of VEGF signaling inhibitors (VSIs). LDH is a tetramer that can include any combination of the M and H subunits. LDH4 and 5 isoenzymes are composed mostly of the M subunit and are more abundant in hypoxic conditions. We speculated that the levels of LDH isoenzymes in pts serum may correlate with outcome of pts treated with VSIs. HORIZON I trial enrolled pts that were randomized to mFOLFOX6 with BEV or CED. A retrospective exploratory analysis was performed on baseline serum samples of these pts. Methods: Total serum LDH was tested on fresh samples during the conduct of the trial. About 2 years after the trial, frozen samples stored at -70 degrees were tested for total serum LDH and LDH isoenzymes, measured by agarose electrophoresis and a colorimetric enzymatic assay. Relative levels of M and H subunits were derived based on the known structure of each isoenzyme. Progression free survival (PFS) and overall survival (OS) were compared by subgroups of total LDH and LDH isoenzyme levels. P values were not calculated for these exploratory analyses. Results: Baseline total LDH levels from fresh serum were available for 207 pts. Total and isoenzyme LDH levels were available for frozen serum samples of 189 pts. Total LDH in the frozen and fresh samples correlated (R=0.9). Distant isoenzymes (e.g. LDH1 and 5) were negatively correlated. High M/H subunits ratio correlated with poor OS (HR=1.804, 95%CI 1.24-2.620). A non-significant trend for better OS to CED-treated vs. BEV-treated pts was seen in pts with high M/H ratio (e.g., CED 30mg vs BEV HR=0.685, 95%CI 0.382-1.23). Conclusions: Evaluation of LDH isoenzymes is feasible using serum samples kept frozen for 2 years. A negative correlation is seen between hypoxic-related and oxic-related isoenzymes. In CRC pts treated with a VSI, LDH isoenzyme hypoxia-associated profile correlates with poorer outcome. LDH isoenzyme profile as a possible predictive biomarker for benefit from CED vs. BEV requires further investigation.
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6

Pompeu, Georgia Bertoni, Priscila Lupino Gratão, Victor Alexandre Vitorello e Ricardo Antunes Azevedo. "Antioxidant isoenzyme responses to nickel-induced stress in tobacco cell suspension culture". Scientia Agricola 65, n. 5 (2008): 548–52. http://dx.doi.org/10.1590/s0103-90162008000500015.

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Exposure to nickel (Ni) at high concentrations can lead to production of reactive oxygen species (ROS) resulting in oxidative damage at the cellular level. We investigated the antioxidative responses of Nicotiana tabacum cv BY-2 cell suspension to Ni stress (0.075 and 0.75 mM NiCl2) over a 72 h period with special attention to potential alterations in isoenzymes of superoxide dismutase (SOD), catalase (CAT) and glutathione reductase (GR). Two main SOD isoenzymes were observed, a Mn-SOD (band I) and a Fe-SOD (band II), as well as one CAT isoenzyme and four GR isoenzymes. Activity staining analysis revealed that CAT activity plays a major role in the early response to Ni-induced oxidative stress, particularly when the Ni concentration used was low, whilst a specific GR isoenzyme appears to respond to the Ni-induced oxidative stress when a much higher Ni concentration was used to induce the stress for the same period of treatment. These results illustrate the importance and advantages of determining individual isoenzyme activities.
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7

Kochhar, Sunita, Christopher B. Watkins, Patricia L. Conklin e Susan K. Brown. "A Quantitative and Qualitative Analysis of Antioxidant Enzymes in Relation to Susceptibility of Apples to Superficial Scald". Journal of the American Society for Horticultural Science 128, n. 6 (novembre 2003): 910–16. http://dx.doi.org/10.21273/jashs.128.6.0910.

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The activities and isoenzyme patterns of guaiacol-dependent peroxidase (POX), ascorbate peroxidase (APX), superoxide dismutase (SOD) and catalase (CAT) were studied in yellow- and red-fruited crab apple [Malus (L.) Mill.] selections from a `White Angel' × `Rome Beauty' cross that show differential susceptibility to the physiological storage disorder, superficial scald. There were no consistent relationships between total enzyme activities and scald incidence, high activities of the enzymes being detected in selections with both high and low susceptibilities to scald. However, additional individual isoforms of some antioxidant enzymes were detected in the scald-resistant selections when compared with scald-susceptible selections. In a native gel system, four guaiacol-dependent POX isoenzymes were detected in both yellow and red scald-resistant selections compared with only two in scald-susceptible selections. Similarly, for anodic acidic POX assayed using benzidine, six isoenzymes were detected in both yellow and red scald-resistant selections compared with five in yellow and four in red susceptible selections. Ten SOD isozymes were detected in scald-resistant yellow-fruited selections compared with only five faint bands in scald-susceptible selections, but similar patterns were not detectable for red-fruited selections. Differences in the presence of various isoenzymes for CAT and APX were also detected among the selections, but associations with scald susceptibility were also affected by fruit color or were inconsistent. The presence or absence of individual isoenzymes may be a better indication of scald resistance or susceptibility than the total enzyme activities. Isoenzyme analyses, especially of POX, could be useful to breeders for the early detection of scald resistance/susceptibility in apples.
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8

Singh, S. V., A. Kurosky e Y. C. Awasthi. "Human liver glutathione S-transferase ψ. Chemical characterization and secondary-structure comparison with other mammalian glutathione S-transferases". Biochemical Journal 243, n. 1 (1 aprile 1987): 61–67. http://dx.doi.org/10.1042/bj2430061.

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The isolation and chemical characterization of the anionic human liver glutathione S-transferase (GST) psi (pI 5.5) are described and compared with other GST isoenzymes reported for rat and human. Amino acid compositional analysis, substrate specificity and isoelectric focusing indicated that GST psi is a unique isoenzyme form of GST. Strikingly, however, amino acid sequence analysis of the N-terminal region indicated that GST psi was identical with GST mu in the first 23 amino acid residues reported. It is likely that these two enzyme forms are at least partially structurally related. In order to investigate further the genetic relationship of GST psi to other reported GST isoenzymes, secondary-structure analysis was performed. Despite substantial differences in the N-terminal-region amino acid sequences of some of the GST isoenzymes, the secondary structure of all the isoenzymes is highly conserved at their N-termini. The general uniformity of the secondary structure of this enzyme class at their N-termini strongly indicated that the observed diversity of these isoenzymes probably occurred as a result of a mechanism of gene duplication followed by divergence rather than a mechanism of convergent evolution.
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9

Maekawa, M., K. Sudo, K. Iwahara e T. Kanno. "Lactate dehydrogenase inhibition by immunoglobulin G in human serum." Clinical Chemistry 32, n. 7 (1 luglio 1986): 1347–49. http://dx.doi.org/10.1093/clinchem/32.7.1347.

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Abstract (sommario):
Abstract Low lactate dehydrogenase (LD; EC 1.1.1.27) activity and an abnormal LD pattern in electrophoretograms of LD isoenzymes in the sera of two patients were caused by inhibition of LD by immunoglobulin G. One of these showed inhibitor activity in the serum upon direct analysis, while the other showed activity only after the immunoglobulin was stripped from the LD. As judged from the LD isoenzyme patterns in serum, the LD inhibitor appeared to act against M subunits. However, quantification of binding affinities to each isolated isoenzyme showed that the LD inhibitor had a stronger effect on LD isoenzymes 2 and 3 (H3M1 and H2M2, respectively).
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10

Segil, N., A. Shrutkowski, M. B. Dworkin e E. Dworkin-Rastl. "Enolase isoenzymes in adult and developing Xenopus laevis and characterization of a cloned enolase sequence". Biochemical Journal 251, n. 1 (1 aprile 1988): 31–39. http://dx.doi.org/10.1042/bj2510031.

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Abstract (sommario):
As part of a study of glycolysis during early development we have examined the pattern of expression of enolase isoenzymes in Xenopus laevis. In addition, the nucleotide sequence of a cDNA clone coding for the complete amino acid sequence of one enolase gene (ENO1) in X. laevis was determined. X. laevis ENO1 shows highest homology to mammalian non-neuronal enolase. Analysis of enolase isoenzymes in X. laevis by non-denaturing electrophoresis on cellulose acetate strips revealed five isoenzymes. One form was present in all tissues tested, two additional forms were expressed in oocytes, embryos, adult liver and adult brain, and two further forms were restricted to larval and adult muscle. Since enolase is a dimer, three different monomers (gene products) could account for the observed number of isoenzymes. This pattern of enolase isoenzyme expression in X. laevis differs from that of birds and mammals. In birds and mammals the most acidic form is neuron-specific and there is only one major isoenzyme expressed in the liver. RNAase protection experiments showed the presence of ENO1 mRNA in oocytes, liver and muscle, suggesting that it codes for a non-tissue-restricted isoenzyme. ENO1 mRNA concentrations are high in early oocytes, decrease during oogenesis and decrease further after fertilization. Enolase protein, however, is maintained at high concentrations throughout this period.
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Tesi sul tema "Isoenzymes Analysis"

1

Rena, Neil Graham. "Identification and analysis of phosphodiesterase isoenzymes". Thesis, University of Glasgow, 1998. http://ethos.bl.uk/OrderDetails.do?uin=uk.bl.ethos.390767.

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MEJIA, DE LEON LUIS. "THE GENETICS OF ESTERASE ISOZYMES IN LETTUCE CULTIVARS". Diss., The University of Arizona, 1986. http://hdl.handle.net/10150/183964.

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Abstract (sommario):
Lettuce, in its cultivated form (Lactuca sativa L.), is a crop of economic importance with several characteristics which make it well suited for biochemical and genetic studies. Biochemical traits such as proteins and enzymes have been studied extensively in many plant species and constitute an important experimental approach to physiological, evolutionary, taxonomic and breeding studies. Seed proteins were extracted from an array of lettuce cultivars. Polyacrylamide gel electrophoresis was employed in a comparative analysis of the soluble protein and isozyme characteristics of seed from each cultivar. Some fall-desert, winter-desert, and coastal cultivars were distinguishable based upon variation in soluble proteins or esterase isozymes. Multiple forms of the carboxylic ester hydrolases or esterases have been shown to occur in a wide variety of plants; their role in the plant cell is still however poorly understood. The inheritance of the esterase isozymes was analyzed by a microelectrophoresis technique which enabled the analysis of individual seeds in the progeny from a cross involving two winter-desert cultivars of contrasting banding phenotypes. Two distinct banding patterns were observed in single seeds of these cultivars; their F₁ hybrid showed a summation of parental patterns, the F₂ segregated in a 1:2:1 phenotypic ratio for these esterase patterns, and backcross segregation ratios were 1:1. Banding pattern differences could be accounted for by the segregation of a single gene with codominant action. Quantitative as well as qualitative differences in esterase activity were observed between the parental lines from two different years of production. Some esterase isozymes were developmentally regulated. Molecular weight determination experiments verified the presence of two gene products of 56 and 62.5 Kd.
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陳堅峰 e Kin-fung Chan. "Phylogenetic relationships and genetic diversity detected by rapd and isozyme analysis of crop and weedy species of amaranthus". Thesis, The University of Hong Kong (Pokfulam, Hong Kong), 1996. http://hub.hku.hk/bib/B29803846.

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Chan, Kin-fung. "Phylogenetic relationships and genetic diversity detected by rapd and isozyme analysis of crop and weedy species of amaranthus /". Hong Kong : University of Hong Kong, 1996. http://sunzi.lib.hku.hk/hkuto/record.jsp?B17665450.

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5

Ashari, Ir Sumeru. "Discrimination between citrus genotypes". Title page, contents and summary only, 1989. http://web4.library.adelaide.edu.au/theses/09A/09aa819.pdf.

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Granger, Andrew. "Genetic relationships and pollination studies in sweet cherry (Prunus avium L) /". Title page, table of contents and abstract only, 1995. http://web4.library.adelaide.edu.au/theses/09PH/09phg758.pdf.

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7

King, Timothy L. (Timothy Lee). "Stock and Species Identification of Selected Marine Fishes and Shellfishes Using Allozyme Analysis and Isoelectric Focusing: Implications for Texas Fisheries Management". Thesis, University of North Texas, 1992. https://digital.library.unt.edu/ark:/67531/metadc277919/.

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Allozyme frequencies and general protein patterns were surveyed among selected Texas marine fishes and shellfishes to illustrate the application of biochemical genetic techniques to stock and species identification in fisheries management.
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Mauricio, Isabel Larguinho. "Genetic diversity in the Leishmania donovani complex". Thesis, London School of Hygiene and Tropical Medicine (University of London), 2000. http://researchonline.lshtm.ac.uk/682281/.

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The Leishmania donovani complex comprises four described species: L. donovani, L. archibaldi, L. infantum and L. chagasi. L. chagasi is the only New World species and has been considered similar to L. infantum, although some authors insist on maintenance of its independent species status. L. donovani has at least two major epidemiological subgroups whose relationships are poorly understood. In this thesis, molecular biological techniques were used to investigate the taxonomy and phylogenetic relationships within the L. donovani complex, with isoenzyme analysis (lEA) as reference technique. Random amplification of polymorphic DNA (RAPD) was used to provide anonymous genetic markers which allowed overall comparisons of genomes. Selected target genes and intergenic regions were also amplified by the polymerase chain reaction (PCR), namely the major surface protease (msp or gp63), the mini-exon and the ribosomal internal transcribed spacer (ITS). PCR products of intergenic regions between msp genes (ITG/CS and ITG/L), mini-exon and ITS were analysed by restriction fragment length polymorphism (RFLP). Phylogenies generated from each of the methods were compared with that of IEA. L. infantum and L. chagasi were found to be synonymous, whilst L. donovani was found to be more polymorphic than L. infantum and a fourth possible species in the complex, L. archibaldi, was not supported. Six genetic groups of strains were identified in the L. donovani complex, based on all DNA based analyses, which agreed with IEA typing. Pooled data from RFLP and RAPD analyses generated robust phylogenies which were congruent with ITG/CS RFLP and msp DNA sequence based phylogenies, but not with lEA phylogenies. The evolutionary history of the L. donovani complex is analysed in the light of the present results. The diverse typing methods were also evaluated and genetic markers suggested, that are applicable to classification and typing of L. donovani species and strains.
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Rowe, Daniel C. "Analysis of Toll-Like Receptor 4 Signal Transduction and IRF3 Activation in the Innate Immune Response: A Dissertation". eScholarship@UMMS, 2006. https://escholarship.umassmed.edu/gsbs_diss/163.

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Over the last decade, the innate immune system has been the subject of extensive research. Often overlooked by the robustness and specificity of the adaptive immune system, the innate immune system is proving to be just as complex. The identification of several families of pattern recognition receptors (PRRs) has revealed an ancient yet multifaceted system of proteins that are responsible for initiating host defense. A wide array of pathogens, from virus to bacteria, is detected using this assortment of receptors. One such family, the Toll-like receptors (TLRs), has been at the forefront of this research. To date, 10 TLRs have been described in the human genome. Activation of TLRs leads to the induction of immune-related genes that ultimately control the response of the host. However, the signaling pathways emanating from activated TLRs and other PRRs are not fully understood. In particular, the pathway leading to the activation of interferon regulatory factor 3 (IRF3), a transcription factor crucial for the induction of type I interferon, remains undefined. IRF3 activation occurs as the consequence of viral infection and through the activation of TLRs 3 and 4 by dsRNA and lipopolysaccharide (LPS), respectively. The focus of this research is to describe components of the IRF3 activation pathway, partly through the analysis of TLR signal transduction. IRF3 normally resides in the cytoplasm of cells. Upon infection with certain viruses and bacteria, IRF3 is activated though phosphorylation at its C-terminus. Phosphorylated IRF3 homodimerizes and associates with co-activators CBP-p300. After translocating to the nucleus, the activate IRF3 complex induces the activation of type 1 interferon and interferon related genes. Little is known about the pathways that lead to the activation of IRF3, especially the kinases involved. In this study we report that the non-canonical IкB kinase homologues, IкB kinase epsilon (IKKε) and TANK-binding kinase-1 (TBK1), which were previously implicated in NF-кB activation, are also essential components of the IRF3 signaling pathway. In particular, mouse embryonic fibroblasts from TBK1 deficient mice fail to activate IRF3 in response to both viral infection and stimulation with LPS or poly (IC), a dsRNA analog. Thus, both IKKε and TBK1 play a critical role in innate immunity and host defense. In addition to viral infection, IRF3 activation also occurs via the activation of TLR3 and 4. TLRs signal through a subfamily of Toll-IL-1-Resistance (TIR) domain containing adapter molecules. One such adapter, MyD88, is crucial for all TLRs, with the exception of TLR3. MyD88 participates in a signal transduction pathway culminating in the activation of the transcription factor NF-кB. Studies from MyD88-deficient mice reveal that both TLR3 and 4 still are capable of activating NF-кB, although with slightly delayed kinetics. Another aspect of the MyD88-independent signal transduction pathway is the activation of IRF3. A second TIR domain containing adapter molecule called Mal/Tirap was discovered and originally thought to mediate the MyD88-independent pathway. However, Mal-deficient mice were found to be defective in both TLR2 and 4 mediated NF-кB activation. We hypothesized that other TIR domain containing adapters could mediate this MyD88-independent pathway of TLR3 and 4 leading to the activation of IRF3. Two additional TIR adapters were discovered, TRIF and TRAM. TRIF was shown to mediate TLR3 signal transduction. In this study, we report that both TRIF and TRAM mediate the activation of the MyD88-independent pathway in response to LPS/TLR4 activation. Unlike any of the other known TIR domain containing adapters, TRAM appears to be restricted to the LPS/TLR4 activation pathway while TRIF plays a role in both TLR3 and TLR4 pathways leading to IRF3 target gene expression. Our studies revealed that TRAM could be acting upstream of TRIF in the LPS/TLR4 pathway. To this end, we sought to determine the localization of TRAM within the cell. We found that TRAM localizes to the plasma membrane. TRAM localization is the result of myristoylation since mutation of the predicted myristoylation site (G2A) resulted in the re-distribution of TRAM from the membrane into the cytoplasm. Reconstitution of TRAM-deficient macrophages with TRAM G2A is unable to rescue LPS/TLR4 signal transduction. Thus, myristoylation and membrane association of TRAM are critical for LPS/TLR4 signal transduction. The data generated in this dissertation extends our understanding of the signaling pathways of the innate immune system. Indeed, the molecules and pathways described herein could prove to be beneficial targets for ameliorating symptoms of disease, both autoimmune and pathogen-associated. Finally, the research described here will spur further insight into the complex signaling pathways of a once ignored arm of the immune system.
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Kwitshana, Zilungile L. "In vitro culture and isoenzyme analysis of giardia lamblia". Thesis, 1999. http://hdl.handle.net/10413/8226.

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Abstract (sommario):
Giardia lamblia, an enteric protozoan parasite, infects a large number of individuals worldwide. In South Africa prevalences ranging between 4 and 63% are documented, however, the impact of giardiasis is underreseached in this country. Giardia infections vary from asymptomatic carriage or a self-limiting acute symptomatic illness to chronic, debilitating malabsorption syndrome. The factors responsible for development of symptomatic versus asymptomatic infection are poorly understood. It is believed by some that host factors determine the clinical outcome of infection. On the other hand, the possibility of the existence of pathogenic and non-pathogenic strains (a situation akin to Entamoeba spp.) remains to be explored. One requirement for investigation of the potential contribution of strain differences to pathogenecity of infection is establishment of laboratory cultures of different strains isolated from symptomatic and asymptomatic patients. The present study was undertaken to develop and modify existing methods for: (i) establishment of laboratory cultures of Giardia trophozoites from excystation of faecal cysts, (ii) long-term maintenance and cryopreservation of the cultures and (iii) preliminary characterisation methodology. One thousand and twenty-three stool specimens were collected from day care centres, hospital wards and Hlabisa hospital laboratory. A further 6246 were retrieved from the Microbiology Laboratory at King Edward VIII Hospital and screened by direct wet preparation. Giardia was detected by light microscopy following formol-ether concentration (127 of 1023 samples) or direct examination of wet preparations (78 of 6246 samples). Cysts were purified from the positive specimens by sucrose gradient separation. Viability was assessed by a dye-exclusion method (eosin). Three in vitro excystation techniques were employed in an attempt to obtain trophozoites for initiation and establishment of viable cultures thereof. Culture conditions were optimised using two reference strains of Giardia, WB & H7 (obtained from the National Institutes of Health, USA). The percentage excystation ranged between 0-42% with all the in vitro methods of excystment. Excysted trophozoites remained viable in TYI-S-33 culture medium for periods ranging between 12-72 hours or up to 9 days, and gradually died, hence viable trophozoite cultures could not be established. Some culture initiates (overall 65%) were lost through overwhelming bacterial and!or fungal contaminants. An animal model was subsequently set up in which C57BL/6 and Praomys (Mastomys) coucha mice were used for in vivo excystation experiments. 1-3 day old suckling mice were intragastrically injected with 10,5 -cysts/ ml in 0,1 ml distilled water. Trophozoites were retrieved from the stomachs of infected mice 7-10 days after inoculation and cultivated in TYI-S-33 medium. Six local isolates were axenised using the in vivo excystation method. They have been maintained for more than 15 months in culture after stabilates and Iysates of confluent growths had been cryopreserved in Liquid Nitrogen. Successful (100%) retrieval of the cryopreserved cultures has been achieved. Seven isoenzyme electrophoresis systems have been set up and optimised. Reproducible results were obtained in six of the enzymes. Some differences in banding patterns of the enzymes were demonstrated.
Thesis (M.Med.Sc.)-University of Natal, Durban, 1999.
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Libri sul tema "Isoenzymes Analysis"

1

Izmenchivostʹ izofermentnykh spektrov peroksidazy u sosny obyknovennoĭ. Sverdlovsk: Akademii͡a︡ nauk SSSR, Uralʹskiĭ nauch. t͡s︡entr, 1985.

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2

Interpretation of protein and isoenzyme patterns in body fluids. New York: Igaku-Shoin, 1991.

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3

Menendez, Ricardo A. Identification of apple (Malus domestica Borkh.) clones based on isozymic diversity. 1985.

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4

Practical isozyme genetics. Chichester: Ellis Horwood, 1988.

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5

Biochemical and immunological analyses of creatine kinase isoenzymes from human heart and skeletal muscle. 1988.

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Biochemical and immunological analyses of creatine kinase isoenzymes from human heart and skeletal muscle. 1988.

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Biochemical and immunological analyses of creatine kinase isoenzymes from human heart and skeletal muscle. 1988.

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Biochemical and immunological analyses of creatine kinase isoenzymes from human heart and skeletal muscle. 1986.

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Serum creatine kinase (CK) and CK-MB isoenzyme responses to acute and prolonged swimming in trained athletes. 1985.

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Capitoli di libri sul tema "Isoenzymes Analysis"

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Millar, Thomas J., e Harry Koutavas. "Analysis of Phosphodiesterase Isoenzymes in the Ocular Glands of the Rabbit". In Lacrimal Gland, Tear Film, and Dry Eye Syndromes 2, 153–56. Boston, MA: Springer US, 1998. http://dx.doi.org/10.1007/978-1-4615-5359-5_21.

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2

Katoch, Rajan. "Isoenzyme Analysis". In Analytical Techniques in Biochemistry and Molecular Biology, 227–31. New York, NY: Springer New York, 2011. http://dx.doi.org/10.1007/978-1-4419-9785-2_11.

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3

Kuan, Shia S., e George G. Guilbault. "Immunochemical Determination of Creatine Kinase Isoenzyme-MB". In Electrochemical Sensors in Immunological Analysis, 145–65. Boston, MA: Springer US, 1987. http://dx.doi.org/10.1007/978-1-4899-1974-8_11.

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4

Dardé, M. L., B. Bouteille e M. Pestre-Alexandre. "Study of Genetic Polymorphism of Toxoplasma Gondii Through Isoenzyme Analysis". In Toxoplasmosis, 9–17. Berlin, Heidelberg: Springer Berlin Heidelberg, 1993. http://dx.doi.org/10.1007/978-3-642-78559-7_2.

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Sen, Asok K. "Control Analysis of Isoenzymic Pathways: Application of Directed Graphs and Electrical Analogs". In Modern Trends in Biothermokinetics, 237–42. Boston, MA: Springer US, 1993. http://dx.doi.org/10.1007/978-1-4615-2962-0_38.

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Gramiccia, M., L. Gradoni e M. C. Angelici. "Epidemiology of Mediterranean Leishmaniasis Caused by Leishmania Infantum: Isoenzyme and kDna Analysis for The Identification of Parasites from Man, Vectors and Reservoirs". In Leishmaniasis, 21–37. Boston, MA: Springer US, 1989. http://dx.doi.org/10.1007/978-1-4613-1575-9_3.

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Dumas-Gaudot, E., B. Dassi, S. Slezack, V. Gianinazzi-Pearson e S. Gianinazzi. "Targeted Approaches for Detecting Changes in Protein Expression with Mycorrhiza Development: Hydrolytic Isoenzyme Analyses and Immunological Detection of Known Proteins in Root Extracts". In Mycorrhiza Manual, 289–309. Berlin, Heidelberg: Springer Berlin Heidelberg, 1998. http://dx.doi.org/10.1007/978-3-642-60268-9_19.

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Gardner, Natwaine Sherune, Kedon J. S. Luke, Andrew O. Wheatley, Winston De La Haye, Perceval Steven Bahado-Singh, Lowell L. Dilworth, Donovan A. McGrowder et al. "Plasma Cocaine Metabolite Levels and Liver CYP450 3A4 Isoenzyme Activity as Indicators of Cocaine Metabolism in Rats Treated With Salako Supplements". In Strategic Applications of Measurement Technologies and Instrumentation, 1–21. IGI Global, 2019. http://dx.doi.org/10.4018/978-1-5225-5406-6.ch001.

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The effects of Salako nutritional supplements on cocaine-dependent Sprague Dawley rats was investigated. Rats were made cocaine-dependent using conditioned place preference (CPP) where craving was analyzed regularly. Cocaine metabolite levels were determined from blood samples. CYP450 3A4 isoenzyme activities were obtained using liver homogenate. Statistical analysis was done using SPSS one-way ANOVA and Duncans multiple range test. Results show that when cocaine use was discontinued, the supplements reduced craving of cocaine significantly. Blood plasma results showed higher benzoylecgonine equilibrium possibly indicating that the supplements aided the removal of stored cocaine metabolites which may have contributed to better management of craving in the rats. CYP450 3A4 isoenzyme activity was further enhanced by the supplements and is indicative of increased cocaine metabolism. The results indicate that the Salako nutritional supplements reduce craving caused by chronic cocaine administration by increasing the liver CYP450 3A4 isoenzyme activity, resulting in better plasma clearance.
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K., Malgorzata. "Isoenzyme Analyses Tools Used Long Time in Forest Science". In Electrophoresis. InTech, 2012. http://dx.doi.org/10.5772/45756.

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Tenor, Hermann, e Christian Schudt. "Analysis of PDE Isoenzyme Profiles in Cells and Tissues by Pharmacological Methods". In Phosphodiesterase Inhibitors, 21–40. Elsevier, 1996. http://dx.doi.org/10.1016/b978-012210720-7/50004-6.

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Atti di convegni sul tema "Isoenzymes Analysis"

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Staykova, Teodora, Panomir Tzenov, Yolanda Vasileva, Dimitar Grekov e Krasimira Avramova. "POPULATION GENETIC ANALYSIS OF SILKWORM BREEDS BASED ON ISOENZYME MARKERS". In XIth International Congress of Geneticists and Breeders from the Republic of Moldova. Scientific Association of Geneticists and Breeders of the Republic of Moldova, Institute of Genetics, Physiology and Plant Protection, Moldova State University, 2021. http://dx.doi.org/10.53040/cga11.2021.111.

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Rapporti di organizzazioni sul tema "Isoenzymes Analysis"

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Watkins, Chris B., Susan Lurie, Amnon Lers e Patricia L. Conklin. Involvement of Antioxidant Enzymes and Genes in the Resistance Mechanism to Postharvest Superficial Scald Development. United States Department of Agriculture, dicembre 2004. http://dx.doi.org/10.32747/2004.7586539.bard.

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The objective of this research project was to evaluate the involvement of antioxidant enzymes and genes in the resistance mechanism to postharvest superficial scald development using two primary systems: 1. Resistant and susceptible progenies of an apple cross between a scald resistant crab apple, ‘White Angel’ and a scald susceptible cultivar, ‘Rome Beauty’; 2. Heat-treatment of ‘Granny Smith’, which is known to reduce scald development in this cultivar. In 2002 we asked for, and received (October 14), permission to revise our initial objectives. The US side decided to expand their results to include further work using commercial cultivars. Also, both sides wanted to include an emphasis on the interaction between these antioxidant enzymes and the á-farnesene pathway, with the cooperation of a third party, Dr. Bruce Whitaker, USDA-ARS, Beltsville. Background: Superficial scald is a physiological storage disorder that causes damage to the skin of apple and pear fruit. It is currently controlled by use of an antioxidant, diphenylamine (DPA), applied postharvest by drenching or dips, but concern exists about such chemical usage especially as it also involves application of fungicides. As a result, there has been increased emphasis on understanding of the underlying mechanisms involved in disorder development. Our approach was to focus on the oxidative processes that occur during scald development, and specifically on using the two model systems described above to determine if the levels of specific antioxidants and/or antioxidant enzyme activities correlated with the presence/absence of scald. It was hoped that information about the role of antioxidant-defense mechanisms would lead to identification of candidate genes for future transgenic manipulation. Major conclusions, solutions, achievements: Collectively, our results highlight the complexity of superficial scald developmental processes. Studies involving comparisons of antioxidant enzyme activities in different crab apple selection, commercial cultivars, and in response to postharvest heat and 1-methylcyclopropene (1-MCP) treatments, show no simple direct relationships with antioxidant contents and susceptibility of fruit to scald development. However, a correlative relationship was found between POX activity or isoenzyme number and scald resistance in most of the studies. This relationship, if confirmed, could be exploited in breeding for scald resistance. In addition, our investigations with key genes in the á-farnesenebiosynthetic pathway, together with antioxidant processes, are being followed up by analysis of exposed and shaded sides of fruit of cultivars that show different degrees of scald control by 1-MCP. These data may further reveal productive areas for future research that will lead to long term control of the disorder. However, given the complexity of scald development, the greatest research need is the production of transgenic fruit with down-regulated genes involved in á- farnesene biosynthesis in order to test the currently popular hypothesis for scald development.
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