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 investigational use of prostaglandins to establish a safe, alternative method for the termination of pregnancy has shown significant development in the United States. The introduction of second generation compounds was initiated by chemically attaching a methyl group in the 15 carbon position of prostaglandins E2 and F2alpha. These compounds prevented enzymatic degradation by the enzyme prostaglandin 15 dehydrogenase. (15S)-15 methyl prostaglandin E2 methyl ester administered by intramuscular injection has been used successfully to therapeutically terminate pregnancy in 208 women of gestational age six through 20 weeks. Side effects, not major and considered acceptable by the investigator, were vomiting, diarrhea and temperature elevations associated with shaking and chills. (15S)-15 methyl prostaglandin F2alpha (THAM), administered by intramuscular injection, has been used to terminate pregnancy in 283 women. Efficacy rates under optimal dosage regimens have reached 100% with a complete abortion rate of 96%. Gastrointestinal side effects of vomiting and diarrhea occurred, but temperature elevations with associated shaking and chills were infrequent. The mean time from initial therapy to abortion with both compounds has remained under 16 hours. A route of drug therapy for therapeutic termination of human pregnancy has been explored and developed which avoids invasion of the uterus.
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PMID:The termination of human pregnancy with prostaglandin analogs. 121 55

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 aim of this work was to increase sensitivity in the detection of antigens from HIV-infected patients, through a process of immune complex dissociation without loss of antigenicity. 500 microliters of sera were mixed with 100 microliters of PEG 12%, stored one night in refrigerator, and centrifuged at 2000 g during 20 minutes. 200 microliters of buffer AcH/Ac- (pH 3.5) were added to the sediment, and incubated at 37 degrees C during one hour with periodic shaking. This was neutralized with 100 microliters of buffer TRIS/CIH (pH 8.6). The antigen was investigated in the original sample, supernatant and sediment. Samples of 105 patients with positive serology, confirmed by Western Blot following CDC criteria, were processed. The antigen was detected in 62 (59%) samples precipitated with PEG, but only 35 (33%) when conventional methods were used. Applying statistics X2: 13.97, P < 0.001, a highly significant association can be observed between PEG dissociation treatment and antigen detection. 27 negative sera by the standard method became positive in the whole sediment, and only 8 in the supernatant. In addition, 40 negative sera were processed, which had not become positive for the antigen by PEG treatment.
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PMID:[Detection of human immunodeficiency virus antigen both free and in immune complexes]. 858 50

Hair pigmentation is a critical factor in the interpretation of the concentration of certain compounds and their metabolites incorporated into hair. Melanin is responsible for the pigmentation. The color and the melanin content of human hair samples differs over a wide range. Once deposited into hair, drug may remain detectable for a period of months to years. However, if drug disposition into hair is influenced by those properties attributed to hair color, then certain persons may test positive more frequently than other persons. Removal of the melanin from hair digests prior to drug analysis may reduce the effect of melanin on the total drug concentration by excluding the drug bound to the pigment. In this study, the effect of melanin removal by centrifugation of hair digests on cocaine concentrations was investigated. Two sets of hair samples from five cocaine users were analyzed for cocaine and metabolites. A solution consisting of 10 mL of 0.5M Tris buffer (pH 6.4) to which is added 60 mg D,L-dithiothreitol, 200 mg SDS, and 200 U Proteinase K, was used to digest the hair. Two milliliters of this solution was added to 20 mg of hair and incubated at 37 degrees in a shaking water bath (90 oscillations/min) overnight. The samples were removed from the water bath and mixed. One set was centrifuged at 2000 rpm and divided into supernatant and melanin pellet. The other set was not centrifuged. Internal standards were added to all tubes. The samples were further extracted, derivatized, and analyzed by gas chromatography-mass spectrometry. A mean of 8.8% (standard deviation [SD] 7.0%) of the total cocaine concentration (supernatant and pellet) was left behind in the pellet. The same experiment was repeated except that the melanin pellet was redigested with 0.1 N HCl. After redigestion of the melanin pellet, the mean cocaine concentration in the pellet was 3.8% +/- 4.0% (mean +/- SD) of the total cocaine concentration in hair. These data demonstrate that removal of melanin from hair digests by centrifugation does not eliminate hair color bias when interpreting cocaine concentrations.
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PMID:Quantitation of cocaine in human hair: the effect of centrifugation of hair digests. 978 14

Glutathione peroxidase (GSH-Px), from commercial bovine erythrocytes or ammonium sulfate fractionations (30-45%, 45-60%, 60-75% and 75-90% saturations) of ginger rhizome, was detected on polyacrylamide gels after native polyacrylamide gel electrophoresis (PAGE) or sodium dodecyl sulfate (SDS)-PAGE. The gel was submerged in a 50 mM Tris-HCl buffer (pH 7.9) containing 13 mM glutathione and 0.004% hydrogen peroxide with gentle shaking for 10-20 min. The GSH-Px activity was stained with a solution containing 1.2 mM 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and 1.6 mM phenazine methosulfate (PMS) for 10 min. The clear zone of GSH-Px activity on a purple background was found in both native and SDS-PAGE gels. This fast and sensitive method can be used in the process of enzyme purification and characterization of mammalian or plant cells.
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PMID:Activity staining of glutathione peroxidase after electrophoresis on native and sodium dodecyl sulfate polyacrylamide gels. 1187 Jul 57

A liquid chromatography method was developed for the determination of antifungal/antimicrobial proteins Rs-AFP1 and Dm-AMP1 in sandy loam soils. The extraction of these highly basic proteins was achieved by mechanical shaking with aqueous Tris buffer pH 9 containing guanidinium thiocyanate salt (4.1 M), EDTA and nonionic polyoxyethylene 20 cetyl ether, Brij-58 detergent. The extracts were cleaned up on Oasis HLB polymer solid-phase extraction cartridges and quantified by liquid chromatography fluorescence detection based on the fluorescence properties of the tryptophan content of these proteins. The detector response was linear for 0.3-10 microg mL(-1). Procedural recoveries were tested in the range 10-100 mg kg(-1). The limit of quantification was 10 mg kg(-1 )protein in the soil sample representing the lowest validated fortification level. The antifungal proteins were found to be stable in soil extract tested up to 9 days when stored at 4 degrees C.
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PMID:Determination of antifungal proteins in soil by liquid chromatography. 1276 64

A series of experiments on various factors which induce optimal in vitro excystation of the metacercariae of Metagonimus yokogawai isolated from the fish, Plecoglossus altivelis was conducted and the following results were obtained. 1)The metacercariae used in this experiment were isolated by the digestion technique therefore all of them were pretreated with the acid-pepsin solution before being applied to the various tests. 2)No excystation occurred when the metacercariae were placed in a salt solutions such as physiological saline, Tyrode solution and Veronal, Tris buffers alone or in combination. 3)The metacercariae underwent complete excystation in the trypsin and pancreatin solution in Tris buffer within an hour at 38 degrees C. The best results were obtained in 0.8-0.9% trypsin solutions, pH 8.0-8.6 and at 38-40 degrees C, approximately one hundred per cent excystation occurred in 40 minutes. Not only temperature but also hydrogen ion concentration played an important role causing excystation of the metacercariae in trypsin-Tris buffer solution. However, bile salts were not responsible for the excystation. 4)Agitation effect on the excystation was tested as a mechanical stimulus and it was found that the shaking stimulus accelerated the excysting mechanism, compared with the metacercariae on which it was not imposed. It is concluded that the metacercariae pretreated in the acid pepsin solution demonstrates an essential requirement for the enzyme solution such as trypsin or pancreatin, provided with the optimum conditions of temperature and hydrogen ion concentration in excysting medium.
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PMID:Study On Metagonimus Yokogawai (Katsurada, 1912) In Korea: II. The In Vitro Excystation Of Metacercariae. 1291 12

We attempted to cryopreserve spermatozoa from closed colonies (Jcl:SD and Jcl:Wistar), and inbred (BN/Crj, F3441 DuCrj, LEW/Crj, Long-Evans and WKY/NCrj), mutant (Zitter [WTC.ZI-zi] and Tremor [TRM]), transgenic (human A-transferase [A], and green fluorescent protein [GFP]) strains of rats. Rat epididymal spermatozoa suspended in cryopreservation solution (23% egg yolk, 8% lactose monohydrate, and 0.7% Equex Stm, pH 7.4, adjusted with 10% Tris [hydroxymethy] aminomethane) were frozen and stored at -196 degrees C. After thawing at 37 degrees C, the spermatozoa were instilled into the tip of each uterine horn of the recipients. A total of five recipient females for each strain were inseminated with cryopreserved spermatozoa, and normal live offspring of all strains (Jcl:SD: 11, Jcl:Wistar: 13, BN/Crj: 9, F344/DuCrj: 28, LEW/Crj: 4, Long-Evans: 6, WKY/NCrj: 8, TRM: 24, WTC.ZI-zi: 27, A: 30 and GFP: 20) were obtained.
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PMID:Cryopreservation of spermatozoa from closed colonies, and inbred, spontaneous mutant, and transgenic strains of rats. 1472 12

With the new concept of 'individualized treatment and targeted therapies', tumour tissue-associated biomarkers have been given a new role in selection of cancer patients for treatment and in cancer patient management. Tumour biomarkers can give support to cancer patient stratification and risk assessment, treatment response identification, or to identifying those patients who are expected to respond to certain anticancer drugs. As the field of tumour-associated biomarkers has expanded rapidly over the last years, it has become increasingly apparent that a strong need exists to establish guidelines on how to easily disintegrate the tumour tissue for assessment of the presence of tumour tissue-associated biomarkers. Several mechanical tissue (cell) disruption techniques exist, ranging from bead mill homogenisation and freeze-fracturing through to blade or pestle-type homogenisation, to grinding and ultrasonics. Still, only a few directives have been given on how fresh-frozen tumour tissues should be processed for the extraction and determination of tumour biomarkers. The PathoBiology Group of the European Organisation for Research and Treatment of Cancer therefore has devised a standard operating procedure for the standardised preparation of human tumour tissue extracts which is designed for the quantitative analysis of tumour tissue-associated biomarkers. The easy to follow technical steps involved require 50-300 mg of deep-frozen cancer tissue placed into small size (1.2 ml) cryogenic tubes. These are placed into the shaking flask of a Mikro-Dismembrator S machine (bead mill) to pulverise the tumour tissue in the capped tubes in the deep-frozen state by use of a stainless steel ball, all within 30 s of exposure. RNA is isolated from the pulverised tissue following standard procedures. Proteins are extracted from the still frozen pulverised tissue by addition of Tris-buffered saline to obtain the cytosol fraction of the tumour or by the Tris buffer supplemented with the non-ionic detergent Triton X-100, and, after high-speed centrifugation, are found in the tissue supernatant. The resulting tissue cell debris sediment is a rich source of genomic DNA.
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PMID:European Organisation for Research and Treatment of Cancer (EORTC) Pathobiology Group standard operating procedure for the preparation of human tumour tissue extracts suited for the quantitative analysis of tissue-associated biomarkers. 1732 Nov 28

The measurement of airborne endotoxins is thus far not standardized. Earlier studies reported higher endotoxin yields when Tween 20 was added to the media used for filter extraction and in the Limulus amebocyte lysate (LAL) assay. This study compared four common media and assessed the effects of Tween during extraction and analysis separately. Parallel airborne dust samples from five work environments (n = 250) were used to compare the four media (pyrogen-free water [PFW], PFW-Tween 20, PFW-Tris, and PFW-triethylamine-phosphate [TAP]) and an extraction time of 10 or 60 min. A subset of the extracts in PFW or PFW-Tween (n = 40) were analyzed in parallel LAL assays with PFW or PFW-Tween as the assay medium. The results produced by a shorter extraction time or the presence of Tris were similar to the results for the reference procedure (PFW and 60 min of shaking). The use of PFW-TAP showed overall lower yields and a deviant calibration curve. The presence of Tween in the extraction medium resulted in significantly (P < 0.05) higher endotoxin yields from all dust types, independent of the effect of Tween in the assay. Tween in the LAL assay, however, also strongly inhibited the reactivity of the lipopolysaccharide (LPS) standard, thus shifting the calibration curve to higher values. The inhibition of LPS in test samples was less pronounced and varied between dust sources, resulting in enhanced calculated concentrations. This assay effect could be circumvented by diluting extracts at least 50-fold before the LAL assay. In conclusion, of the media tested, only Tween enhances the efficiency of endotoxin extraction from airborne dust samples in a consistent manner. We recommend extraction in PFW-Tween combined with dilution and LAL analysis in PFW.
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PMID:Effect of extraction and assay media on analysis of airborne endotoxin. 1844 Nov 12


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