[DISCUSSION ON A SIMPLE METHOD OF ANALYZING METANEPHRINE PLUS NORMETANEPHRINE IN THE DIFFERENTIAL ANALYSIS OF PHEOCHROMOCYTOMA].
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Body Fluids↗
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Cell cycle withdrawal without concomitant differentiation: analysis of a G1-specific temperature-sensitive murine myoblast cell line.
Skeletal muscle differentiation is accompanied by the withdrawal of the proliferating myoblasts from the cell cycle in the G1 phase. We showed earlier that the length of G1 and the timing of the differentiative transition could be controlled in large part by the composition of the culture medium. In this study we have asked whether a G1 arrest imposed independently of the culture medium is sufficient to elicit the differentiative response. To examine this possibility we have characterized a new G1-specific ts murine myoblast line. This line, ts-36, was identified as a G1-specific mutant on the basis of four criteria: prolonged viability at the nonpermissive temperature (npt), the kinetics of cell cycle withdrawal and reentry in temperature shift experiments, the ability of the cells to differentiate at the npt in low-growth medium, and, finally, the observation that, by the criterion of flow microfluorometry, the mutant cells block at the G1 landmark in the cell cycle. A ts-imposed G1 arrest of up to 96-hr duration is by itself insufficient to activate the differentiative program in ts-36 cells cultured in complete growth medium. The differentiated phenotype is expressed, however, in temperature-arrested cells cultured either in low-growth (conditioned) medium or in a medium from which mitogens have been removed by ultrafiltration. Differentiation can be reversed by refeeding with complete growth medium. The effects of growth medium can be mimicked by FGF to the extent of inhibiting activation of the differentiative program in temperature-arrested ts-36 cells and in eliciting downregulation of muscle-specific contractile protein synthesis. Extrapolating from these observations suggests that growth factors may have more than one role in myogenesis in vitro. They not only stimulate proliferation, but also inhibit differentiation in the absence of proliferation. Examining the kinetics of withdrawal from the cell cycle indicates that ts-36, cultured in conditioned medium blocks at the npt restriction point rather than the conditioned medium block. Our results suggest that two conditions must be met to trigger myogenic differentiation in vitro. Withdrawal from the cell cycle in G1 alone is not sufficient. Reduction of the mitogen level in the medium below a threshold level is an obligate condition for phenotypic expression.
Animals↗
[Cloning, location and differential analysis of transcription factor Relish gene from Anopheles stephensi under different feeding conditions].
OBJECTIVE: To clone, locate and differentially analyze the transcription factor Relish gene from Anopheles stephensi, and to examine its signals-modulating action on prophenoloxidase cascade and melanization of Plasmodium yoelii oocysts. METHODS: Relish cDNA of total mosquitoes was amplified by RT-PCR with degenerated primers. Target PCR product was purified, cloned, sequenced and identified. Special Relish gene was amplified with specific primers from hemocytes or midgut, respectively. Semi-quantitative analysis was made under different feeding conditions. Relish message ribonucleic acid was identified with hybridization in situ. RESULTS: One cDNA segment of Relish similar to An. gambiae was acquired from An. stephensi. The same Relish gene was also manifested in the hemocytes and midgut. Marked up-regulation expression of Relish was observed at 6, 12, 24 or 48 h of Plasmodium yoelii infection and at 12 and 24 h after sucking nitroquine-acetate sucrose solution, that was before inducible oocyst melanization. Relish was also expressed in the hemocytes and midguts by ISH. CONCLUSION: Transcription factor Relish of An. stephensi might play a role in signal modulation of Plasmodium yoelii infection and oocyst melanization.
Amino Acid Sequence↗
[Differential analysis of the binding forms of iron in various plant foodstuffs].
In fourteen plant foodstuffs of various composition a differentiation of iron contents according to water soluble, in ethylacetate extractable, in labile contents and iron(II) or Fe(III) species in the water extract are carried out by the spectrophotometric Ferrozine method. High water-soluble labile iron parts and high iron(II) contents were found in fruit, high complexed parts in the water extract and ethylacetate extractable iron contents in protein rich foodstuffs. The iron total contents are situated between 1 and 90 ppm, the water soluble parts amount to 6 up to 60% of the total content according to the foodstuffs.
Acetates↗
CSF analysis differentiates multiple-system atrophy from idiopathic late-onset cerebellar ataxia.
BACKGROUND: Differentiating idiopathic late-onset cerebellar ataxia (ILOCA) from ataxia due to the cerebellar subtype of multiple-system atrophy (MSA-C) can be difficult in the early stages of the disease METHODS: The authors analyzed the levels of various CSF biomarkers in 27 patients with MSA-C and 18 patients with ILOCA and obtained cut-off points for each potential biomarker to differentiate MSA-C from ILOCA. RESULTS: Increased levels of neurofilament light chain (NFL) and neurofilament heavy chain (NFHp35) and decreased levels of the neurotransmitter metabolites homovanillic acid (HVA), 5-hydroxyindoleaceticacid (5-HIAA), and 3-methoxy-4-hydroxyphenylethyleneglycol (MHPG) were observed in MSA-C compared with ILOCA patients. Receiver operating characteristic analysis showed high sensitivity and specificity levels for NFL, NFHp35, and MHPG analysis. At a cut-off of 24.4 ng/L for the NFL analysis, a sensitivity of 79% and a specificity of 94% were obtained for differentiating MSA-C from ILOCA. At a cut-off point for NFHp35 of 129.5 ng/L, sensitivity was 87% and specificity 83%. Analysis of MHPG levels (cut-off 42.5 nM) resulted in a sensitivity of 86% with a specificity of 75%. A multivariate logistic regression model selected NFL, MHPG, and tau as independent predictive biomarkers that separated the MSA-C and ILOCA groups. CONCLUSIONS: Increased levels of neurofilament light chain and tau and decreased levels of 3-methoxy-4-hydroxyphenylethyleneglycol were associated with high accuracy levels in differentiating the cerebellar subtype of multiple-system atrophy from idiopathic late-onset cerebellar ataxia (LOCA). CSF analysis may thus serve as a useful tool in early diagnostic differentiation of LOCA.
Age of Onset↗
[Differential thermal analysis and differential scanning calorimetry: basis, methods and use].
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Calorimetry↗
[Study of the binary mixtures, khellin-sulfacetamide and khellin-sulfanilamide, by thermomicroscopy, differential thermal analysis, and differential calorimetry].
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Calorimetry, Differential Scanning↗
[The importance of differential analysis of formal thought disorders in confusion psychosis and cataphasia. Incoherence of thematic choice and incoherence with logical and linguistic errors].
Formal thought disorders are usually regarded as a central symptom of schizophrenic psychoses, but nevertheless as diagnostically non-specific. Karl Leonhard, therefore, differentiated in his classification of endogenous psychoses between a whole lot of diagnostically specific types of formal thought disorders. He described two psychoses, of which formal thought disorder is the main psychopathological feature, i.e. confusion psychosis and cataphasia. By means of an analysis of tape-recorded utterances of patients with such psychoses we demonstrate the phenomenological differentiation of the distinct types of thought disorder in these psychoses. In confusion psychosis the disturbance concerns the thematic organisation and target-oriented direction of the train of thought. Characteristically, an "incoherence of thematic choice" results, reflecting a disturbance on the level of organisation of discourse. The concepts, ideas and themes itself are organised in a logically correct manner and are comprehensible. The thought disorder in cataphasia, however, shows severe logical faults in the connection of concepts and ideas, as a result of which single themes and ideas become a logical and incomprehensible. Moreover, cataphasic thought disorder goes beyond the level of discourse organisation and extends to disturbances of linguistic functions including mistakes on a semantical and syntactic level. These observations correspond well with the widely accepted assumption of a modular organisation of cognitive and linguistic processes. The thought disorder in prognostically unfavourable cataphasia concerns "basal" logical, semantic and syntactic systems, which remain uninfluenced in the prognostically favourable confusion psychosis. In the latter, therefore, the disorder is restricted to the higher functions of organisation of discourse.
Humans↗
Real-time pneumolysin polymerase chain reaction with melting curve analysis differentiates pneumococcus from other alpha-hemolytic streptococci.
The majority of pneumococcal isolates can be identified by the conventional methods based on phenotypic characteristics. Occasionally, however, the differentiation of alpha-hemolytic streptococci from pneumococci, especially those isolated from nasopharynx, is problematic due to the discrepant results obtained by the conventional identification methods. Several gene technological methods based on the amplification of genes encoding pneumococcal virulence factors, such as pneumolysin (ply) and autolysin, have been used as additional identification methods. Recent studies have shown that especially the ply gene is frequently also present in nonpneumococcal alpha-hemolytic streptococci. In this study, we compared the commonly used phenotypic identification methods with nucleic acid-based methods, commercial AccuProbetrade mark, conventional pneumolysin polymerase chain reaction (Ply-PCR), and real-time Ply-PCR in the identification of alpha-hemolytic streptococcal strains isolated from 100 consecutive nasopharyngeal specimens. We also studied if melting curve analysis and sequencing of the amplification products of a ply gene fragment could be helpful in the identification. Our results suggest that the ply gene present in alpha-hemolytic streptococci differs from that present in pneumococcus, and that melting curve analysis would prove useful in the differentiation of these bacteria.
Bacterial Proteins↗
[Differential analysis of two-dimension gel electrophoresis profiles of human lung squamous carcinoma and tumor-adjacent tissue].
BACKGROUND & OBJECTIVE: Carcinogenesis of lung squamous carcinoma is a complex process involving multiple events and steps. Though some molecular pathogenesis studies on human lung cancer have been undertaken successfully in gene (DNA) and transcription (mRNA) levels, the carcinogenic mechanism is still unclear. At present, there is no special molecular marker for early-stage diagnosis and prognosis evaluation. The objective of this study was to establish two-dimensional electrophoresis profiles with high resolution and reproducibility from human lung squamous carcinoma tissue and paired normal tumor-adjacent bronchial epithelial tissue, and to identify differential expression proteins. METHODS: The total proteins of human lung squamous carcinoma tissue and paired normal tumor-adjacent bronchial epithelial tissue were separated by immobilized pH gradient (IPG)-based two-dimensional gel electrophoresis (2-DE). The differential expression proteins were analyzed using image analysis software, then identified using mass spectrometry and database searching. RESULTS: (1) For tumor tissues, the average protein spots of 3 gels were 1349+/-67; and 1235+/-48 spots were matched with the average matching rate of 91.5%. For control, the average protein spots of 3 gels were 1297+/-73; and 1183+/-56 spots were matched with the average matching rate of 91.2%. The average position deviation of matched spots was 0.873+/-0.125 mm in IEF direction, and 1.025+/-0.213 mm in SDS-PAGE direction. (2) A total of 1069+/-45 spots were matched between the electrophoretic maps of 15 human lung squamous carcinoma tissue and paired normal tumor-adjacent bronchial epithelial tissue. Forty differential proteins were identified by peptide mass fingerprint(PMF), some proteins were the products of oncogenes, and the others involve in the regulation of cell cycle and signal transduction. CONCLUSION: In this study, the well-resolved, reproducible 2-DE patterns of human lung squamous carcinoma and adjacent normal bronchial epithelial tissues were established and certain differential proteins were characterized. These data will be helpful for screening the biomarker to further study on human lung squamous carcinoma.
Amino Acid Sequence↗
Do CNS anlagen have plasticity in differentiation? Analysis in quail-chick chimera.
Heterotopic transplantations of brain vesicles of a quail embryo into a chick embryo were carried out in order to elucidate if CNS anlagen have plasticity in differentiation at the 7-10 somite stage. Quail cells are distinguished from chick cells due to the difference in nuclear morphology. The prosencephalon did not differentiate into the cerebellum when transplanted into the metencephalon, although previous study showed that the prosencephalon has the capacity to differentiate into the optic tectum. The mesencephalon differentiated as an optic tectum when transplanted into the prosencephalon or into the rhombencephalon. The metencephalon differentiated as a cerebellum in the telencephalon. It is concluded that only the prosencephalon has limited plasticity, but the mesencephalon and rhombencephalon are determined by the 7-10 somite stage.
Animals↗
X-ray elemental analysis differentiates blood vessels and lymphatic vessels in the chick choroid.
PURPOSE: Chick choroidal lymphatics swell rapidly during recovery from form-deprivation occlusion, leading to the question of functional significance. To date, analysis of lymph in initial lymphatics has been problematic because of the difficulties of access. However, elemental composition can be determined using scanning electron microscope (SEM) X-ray microanalysis. This study investigated whether cryo-fixation would permit vascular fluids to be analysed in situ. METHODS: Two chicks were raised normally and seven were raised with monocular occlusion for 12 days before varying periods of normal visual experience. The eyes were rapidly frozen, fractured and X-ray spectra obtained from the lumina of lymphatic and blood vessels in a Cambridge S360 SEM. RESULTS: The elemental signatures of Na/Cl/K/P/S distinguished the two vessel types. CONCLUSIONS: These results suggest this bulk-frozen tissue technique can quantify relative changes in elemental species present in various ocular compartments in response to light-induced changes in the retina.
Angiography↗
Differentiation analysis of bovine T-lymphosarcoma.
The tumor cells from three different types (calf, skin and thymic) of bovine lymphosarcoma identified as sporadic bovine leukosis (SBL) were transplanted to nude mice and established as lymphoid tumor lines. To clarify the T-cell lineage, clonality and differentiation stage, these cell lines were examined for the immunophenotypic profile, the expression and the gene rearrangement of T-cell receptor (TCR) genes. The tumor cells from all three types showed alpha-naphthyl acetate esterase-positive staining and the absence of surface immunoglobulin (sIg). Moreover, the tumor cells appeared to be all BoCD5+ BoCD6+ BoCD4- BoCD8- cells in the FACS analysis with mAbs. Rearrangement of TCR-beta and -delta, but not of TCR-alpha and -gamma, genes was found to be unique for each tumor line showing T-cell clonality. Mature or truncated TCR-beta, -gamma and -delta transcripts were detected in the tumor cells, but all of them were found to be non-functional in cDNA cloning analysis. These results suggested that tumor cells from SBL are immature double-negative T-cells expressing non-functional TCR gamma delta transcripts to somewhat differentiated degrees.
Amino Acid Sequence↗
Differential analysis of animal bone marrow by flow cytometry.
A simple procedure was developed for rapid analysis of animal bone marrow by flow cytometry using the lipophilic cationic dye 3,3'-dihexyloxacarbocyanine iodide [DiOC6(3)]. The batch process allows differentiation of bone marrow cells into lymphoid, erythroid, and myeloid populations and enables classification of erythroid and myeloid cells into proliferating and maturing subpopulations. From these data, myeloid:erythroid (M:E) ratios and maturation indices for erythroid and myeloid cells (EMI and MMI, respectively) can be derived. This procedure provides the opportunity to analyze bone marrow quantitatively and offers distinct advantages to current manual methods in terms of simplicity, throughput, and reproducibility. The method has been tested successfully using marrow from Wistar rats, B6C3F1 mice, beagle dogs, and cynomolgus monkeys. This technique facilitates the evaluation of bone marrow samples taken from preclinical safety studies or from animal colonies of large size.
Animals↗
T cell receptor-CD3 complex during early T cell differentiation. Analysis of immature T cell acute lymphoblastic leukemias (T-ALL) at DNA, RNA, and cell membrane level.
T cell acute lymphoblastic leukemias (T-ALL) can be regarded as the malignant counterparts of cells in various T cell differentiation stages. To study the expression of the human T cell receptor (TcR)-CD3 complex during the early stages of T cell differentiation, we have analyzed 22 T-ALL at the cell membrane level and the DNA level and 12 of them at the RNA level. According to their immunologic phenotype, the T-ALL could be divided into three main groups: 10 immature T-ALL (CD1-/CD3-), seven common thymocytic T-ALL (CD1+/CD3-or+), and five mature T-ALL (CD1-/CD3+). Among the 10 immature T-ALL three appeared to express the immunologic phenotype of the putative prothymocyte (TdT+/HLA-DR+/CD7+/CD2+/CD5-/CD1-/CD3-), whereas the other seven T-ALL appeared to be immature thymocytic (TdT+/HLA-DR-/CD7+/CD2+/CD5+/CD1-/CD3-). Transcripts of the CD3-delta and CD3-epsilon genes were present in all CD3- and CD3+ T-ALL tested, including prothymocytic T-ALL. However, prothymocytic T-ALL had germline TcR-beta genes and were not rearranged to the characterized TcR-gamma joining regions. The presence of CD3 transcripts and absence of TcR gene rearrangements in prothymocytic T-ALL supports their immature T cell character. Two immature thymocytic T-ALL also had germline TcR-gamma genes and one of them had germline TcR-beta genes. In all other T-ALL the TcR-gamma and TcR-beta genes were rearranged. The presumptive functional 1.3-kilobase TcR-beta transcripts were detected in the majority of T-ALL with rearranged TcR-beta genes. Distinct levels of TcR-gamma transcripts appeared to be present only in some thymocytic T-ALL, i.e., some immature thymocytic T-ALL and common thymocytic T-ALL. TcR-alpha mRNA could only be detected in CD3+ mature T-ALL, but was absent in all CD3+ common thymocytic T-ALL tested. Our data indicate that CD3 gene transcription is one of the earliest events during T cell differentiation and already occurs in prothymocytes. The TcR-gamma and TcR-beta genes rearrange early during thymocytic differentiation and can subsequently be transcribed. High levels of TcR-gamma gene transcription may only occur in a part of the T cells during thymic differentiation, while TcR-beta gene transcription continues during further differentiation. TcR-alpha gene transcription may be the final step in the production of the complete set of TcR and CD3 proteins, resulting in the expression of the TcR alpha beta-CD3 complex at the cell surface of mature T cells.(ABSTRACT TRUNCATED AT 400 WORDS)
Adolescent↗
Pig genome analysis: differential distribution of SINE and LINE sequences is less pronounced than in the human and mouse genomes.
The distribution of SINE and LINE sequences in the pig genome was examined by fluorescence in situ hybridization (FISH), interspersed repeat PCR, and restriction analysis of high molecular weight DNA. FISH revealed a largely uniform hybridization to the euchromatic chromosome regions with both interspersed repeats, although a bias toward the G-bands was observed for the LINE probe. Southern blots of inter-SINE and inter-LINE PCR products showed strong hybridization to LINE and SINE probes, respectively. High molecular weight DNA derived from a pig x hamster hybrid cell line was cut with a panel of G + C and A + T rich rare cutter restriction enzymes, then run on a pulsed field gel and Southern blotted. Sequential hybridization with SINE and LINE probes showed that SINE hybridization was to relatively low molecular weight fragments with the G + C rich enzymes, whereas the LINE probe gave hybridization to significantly larger fragments produced by these enzymes. DNA samples digested with A + T rich enzymes gave essentially similar patterns with SINE and LINE probes. We conclude that the pattern of differential distribution of SINEs and LINEs, which has been described in man and mouse, does exist in the pig but is much less pronounced.
Animals↗
Differential analysis of DNA microarray gene expression data.
Here, we review briefly the sources of experimental and biological variance that affect the interpretation of high-dimensional DNA microarray experiments. We discuss methods using a regularized t-test based on a Bayesian statistical framework that allow the identification of differentially regulated genes with a higher level of confidence than a simple t-test when only a few experimental replicates are available. We also describe a computational method for calculating the global false-positive and false-negative levels inherent in a DNA microarray data set. This method provides a probability of differential expression for each gene based on experiment-wide false-positive and -negative levels driven by experimental error and biological variance.
Analysis of Variance↗
Iron-regulated proteins in Phanerochaete chrysosporium and Lentinula edodes: differential analysis by sodium dodecyl sulfate polyacrylamide gel electrophoresis and two-dimensional polyacrylamide gel electrophoresis profiles.
Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional gel electrophoresis (2-DE) were used to identify iron-responsive proteins in the white-rot species (Phanerochaete chrysosporium and Lentinula edodes), by comparing the differential patterns of cellular and membrane proteins obtained from iron-sufficient and iron-deficient mycelia. Six cellular proteins induced by iron restriction have been observed in SDS-PAGE for P. chrysosporium and twelve for L. edodes. In 2-DE, the numbers of iron-restricted induced proteins were 12 and 9, respectively, in a resolution range of 15-60 kDa and pI 4.5-8.1. SDS-PAGE for the plasma membrane protein did not show differences, whereas the outer-membrane protein profile showed 6 and 5 proteins induced by iron depletion in P. chrysosporium and L. edodes, respectively. The results presented here are important data to unravel mechanisms of biosynthesis and/or transport of the iron-complexing agents in ligninolytic fungi and to further correlate them to the ligninolytic processes.
Chrysosporium↗