Gene/Protein Disease Symptom Drug Enzyme Compound
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Query: UMLS:C0040822 (tremor)
 18,428 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

The frozen tissue was sliced and then homogenized at 20 degree C in LiCl, 2 M; lauryl-trimethyl ammonium chloride (K & K No. 4484), 5%; pronase, B grade (calbiochem), 1 mg/ml and Tris-HCl 10 mM pH 7.5. The homogenate was left to stand 15 min at 20 degree C with occasional shaking. After centrifugation at 35 000 X g for 30 min at 0 degree C, the supernatant containing the crude DNA was purified by filtration on Ultrogel A 2 (LKB, Sweden). The Ultrogel A 2 column (2.5 X 45 cm) was equilibrated with a solution containing NaCl 2 M, EDTA 2.5 mM, Tris-HCl 10 mM pH 7.5. The flow rate was 3 ml cm-2 h-1. Five ml of the supernatant were placed on the column. The first peak contained highly polymerized (as demonstrated by ultracentrifugation) pure DNA (A260/A230 = 3.19; A260/A280 = 1.82). The yield was 2.26 mg of DNA/g of fresh liver.
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PMID:[Simple method for preparation of rat liver DNA]. 617 Dec 20

The ciliated protozoan Colpoda cucullus has been cultivated at 27 degrees C with gentle shaking in a baked lettuce infusion supplemented with Klebsiella suspensions. Under these conditions cells had a mean generation time of about 7 hours and could attain densities up to 20,000/ml and 45,000/ml in the log and stationary phase of growth, respectively. Nuclear preparations obtained from exponentially growing cells by the gum arabic-octanol method showed a satisfactory degree of purity and integrity. They consisted primarily of the large macronuclei attached to which the small micronuclei were sometimes visible. Upon incubation at 27 degrees C in conventional reaction mixtures nuclear preparations actively incorporated 3H-UTP and 3H-dTTP into acid-insoluble material. alpha-amanitin caused a 50% inhibition of RNA synthesis whereas aphidicolin did not affect at all DNA synthesis.
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PMID:[Synthesis of RNA and DNA in isolated nuclei of Colpoda cucullus (Protozoi Ciliati)]. 619 3

A technique is described for the growth of pure gametocyte cultures of Plasmodium falciparum. Using the classification of Hawking, Wilson & Gammage (1971) these cultures contain gametocytes of stages III, IV and V alone. Routine sexual cultures, varying from 0.35 ml static cultures to 500 ml shaking cultures, are exposed to an inhibitor of DNA synthesis (mitomycin C at 10 micrograms/ml) on the 11th day of culture, and the culture is harvested on the 14th day when capacitated stage V gametocytes are present. All other stages are killed by the drug and are morphologically degenerate within 2 days of the addition of the inhibitor.
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PMID:Gametocytogenesis of Plasmodium falciparum in vitro: a simple technique for the routine culture of pure capacitated gametocytes en masse. 637 71

Aqueous acetic acid was used to fix and store specimens of tissue prior to dissociation into nuclear suspensions for flow cytometric quantitation of DNA. The optimum concentration was 20 volumes of glacial acetic acid in 80 volumes of distilled water. Both neoplastic and benign nuclei were easily released from the fixed tissue blocks by slicing and shaking. Residual undissociated tissue was suitable for microscopic examination to confirm its identity. The nuclei fluoresced brightly after staining with propidium iodide, and yielded histograms similar to those from unfixed samples. Acetic-acid fixation resulted in slightly broader G1 and G0 peaks in the DNA histograms in comparison to unfixed cells, but fluorescent debris was less and correlation between the flow cytometric S-phase fraction (SPF) and in vitro thymidine labelling index (TLI) was better than with unfixed cells. Twenty-one of thirty-nine acetic-acid-fixed breast carcinomas had DNA indices in excess of 1.0 (increased nuclear DNA content in comparison to benign cells), and eighteen had DNA indices of 1.0 (normal or near-normal). The SPF was usually in excess of the TLI, but the two were significantly correlated (r = 0.72, P less than 0.0001). However, a significant correlation of SPF with TLI held only for the group with DNA index greater than 1.0. DNA indices greater than 1.0 were associated with high SPF and TLI, and high SPF and TLI each associated with low content of estrogen receptor.
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PMID:DNA flow cytometry of breast carcinoma after acetic-acid fixation. 669 73

Because cytokinetic studies of the human bone marrow aspirate as a prognostic factor and as a monitor of drug perturbation are frequently inconsistent, we investigated reproducibility of DNA distribution measured by flow cytometry of DNA content in patients with morphologically normal bone marrow. In 15 patients, correlation was noted between DNA distributions simultaneously obtained on right and left iliac crest bone marrow aspirates (r = .588), although considerable variation in individuals was encountered. Much better reproducibility (r = .879) was achieved using bilateral core biopsy of bone marrow in these same patients. In 60 samples, comparison of DNA distribution between bone marrow aspirate and simultaneously obtained biopsy revealed higher relative proportions of S and G2 + M phase cells in biopsies (p less than 0.001), suggesting peripheral blood contamination of aspirate material. Brisk shaking of biopsy specimens in saline expelled a representative sample in the supernatant that could be subjected to simultaneous cytomorphological and cytokinetic analysis. To improve reproducibility of DNA content determinations in normal human bone marrow, bone marrow biopsy should be utilized.
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PMID:Flow cytometry of DNA content in human bone marrow: a critical reappraisal. 676 7

Asynchronously growing Chinese hamster cells (M3-1) were UV-irradiated (lambda = 254 nm) and then incubated with/without caffeine (2 mM) for 20 h. Microscopic evaluation of metaphase spreads revealed that after UV-irradiation alone (5.0 J/m2) appearing fragmented and/or pulverized ('GCS-like' cells; GCS, Generalized Chromosome Shattering) was very low while it was high following the combined treatment. Cytogenic and flow cytometric analysis of cells obtained by mechanical shaking cultures treated with UV and caffeine indicated that 'GCS-like' cells have the same DNA content as untreated cells in G2 phase and mitosis.
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PMID:DNA content of cells with generalized chromosome shattering induced by ultraviolet light plus caffeine. 709 88

Chinese hamster ovary (CHO) cell populations were exposed independently to thymidine (TdR) or 5-bromodeoxyuridine (5-BUdR) for either one cell generation (approximately 18 h) or two (36 h) and subsequently synchronized in mitosis either by chemical arrest with colcemid or by mechanical shaking. Upon subsequent X-irradiation at various stages in the cell cycle, the synchronized populations were analyzed for cellular lethality and chromosomal damage. This investigation confirmed that each radiation response was "age-dependent" and essentially similar for both methods of synchrony. Cells irradiated in mitosis were found to be most radio-sensitive while cells irradiated during the DNA synthesis stage were most radio-resistant. A comparative analysis revealed that the two responses of radiation were more pronounced for cells labelled with 5-BUdR than they were for TdR-labelled cells. A further enhancement of radiation responses was noted for cells that were grown in the presence of the halogenated pyrimidine analog for two cell generations, thus re-affirming that the nucleus is a primary target for radiation damage. The comprehensive conclusion sought with regard to the degree of correlation between chromosomal damage and loss of cellular reproduction integrity awaits such information as the molecular organization of mammalian chromosomes and the mechanisms by which aberrations are produced within these chromosomes.
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PMID:The effects of pretreatment of synchronized CHO cells with 5-BUdR and TdR on chromosomal damage and cell death. 733 73

A suction blister technique on the abdominal skin to separate pure human epidermis from its basement membrane is described. After incubation with proteolytic enzymes (trypsin, pepsin) and mechanical shaking, isolated cells and nuclei were stained with ethidium bromide. DNA-specific fluorescence was measured in a flow cytometer using UV excitation light. Cell-cycle phase distributions were obtained from the resulting histograms using a planimetric method. The proportion of cells with an S-phase DNA content showed circadian variations with peak values shortly before noon, and lower values during the rest of the 24 h period.
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PMID:Flow cytometry (FCM) of human epidermal cells. A preparation method for epidermal cells and demonstration of circadian variations in the proportion of S-phase cells. 745 5

Paralytic tremor (pt) is a sex-linked mutation in rabbit that affects myelination of the CNS. Myelin in the pt brains represents approximately 30% of the normal levels. Previously we showed that the pt mutation affects primarily proteolipid protein (Plp) gene expression. In the present study we investigated the relative effect of the pt mutation on two distinctive Plp gene products, PLP- and DM-20-specific messenger RNAs. Our results showed that both PLP and DM-20 are affected and that the ratio DM-20/PLP was higher in pt rabbits than in age-matched controls. We sequenced normal rabbit PLP cDNA and characterized pt mutation at the DNA level. Rabbit PLP sequence, deduced from cDNA, differs from the human protein only at Thr198. Sequence analysis of the mutant cDNA revealed a transversion T-->A in exon 2 of the Plp gene. This point mutation, which is placed at the end of the first potential transmembrane domain, results in a substitution of His36 by a glutamine. This transversion abolishes a restriction site that enabled us to screen a large number of animals and observe a perfect correlation between the pt allele and the abnormal phenotype.
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PMID:Paralytic tremor (pt): a new allele of the proteolipid protein gene in rabbits. 752 75

Inherited neurological diseases in animals are of interest to a wide range of scientific disciplines, particularly because such animals may be suited as genetic animal models for respective human disorders. Because the pig has a number of anatomic and physiologic features similar to those of human beings, this species is becoming increasingly popular in biomedical research. The usefulness of pigs as genetic models of neurological diseases is illustrated by the porcine model of malignant hyperthermia (MH), i.e., a frequently fatal myopathic disease in both pigs and humans. In the present study, we describe a new hereditary movement disorder in Pietrain pigs, which may represent a useful genetic animal model of high-frequency tremor. Because the disorder was first detected in the offspring of a boar named "Campus," we use the term "Campus syndrome" in this respect. Segregation analysis of breeding studies indicates that the syndrome is inherited as a monogenic dominant trait. DNA-based testing of the mutation involved in MH myopathy showed that expression of the Campus syndrome in pigs is not dependent on homozygosity for the MH mutation. In affected pigs, the Campus syndrome develops at an average age of 27 days. The syndrome is characterized by muscular weakness and a very intense tremor of the legs when standing and walking but not when at rest in a lying position. The intensity of tremor and muscular weakness progressively increases with age, resulting in pronounced postural instability. Despite these neurological abnormalities, body weight gain in affected pigs does not differ from that in unaffected siblings.(ABSTRACT TRUNCATED AT 250 WORDS)
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PMID:The "campus syndrome" in pigs: neurological, neurophysiological, and neuropharmacological characterization of a new genetic animal model of high-frequency tremor. 755 40


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