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Biomedical subjects

L Charles

Publications and source records attributed to L Charles.

At least 19 recordsLinked to original sources
PubMed2002

Conversion of dehydroepiandrosterone sulfate at physiological plasma concentration into estrogens in MCF-7 cells.

Metabolism of dehydroepiandrosterone (DHEA), its sulfate (DHEAS), and androstene-3,17-dione (delta(4)) was performed at their physiological plasma concentrations in MCF-7 cell cultures (1 microM, 10 and 2 nM, respectively). Final metabolic products of these steroids were separated by HPLC-radioactive flow detection and identified by LC/MS or MS/MS. Typical and specific mass fragmentation spectra identified the presence of estrone (E(1)), 17beta-estradiol (E(2)), delta(4), DHEA, 5-androstene-3beta,17beta-diol (delta(5)), and testosterone as principal DHEAS metabolites. Other steroids, such as androstenedione, androsterone, and DHEA fatty acid esters at very low concentrations (from pM to nM), were also obtained after steroid incubation. This highly specific method allowed us to conclude whether a metabolite and enzymatic activity of interest were present in MCF-7 cells or not. We also showed that DHEAS at its physiological plasma concentration may be converted into estrogens and estrogen-like compounds in breast cancer cells. The estrogenic action of DHEAS on breast cancer cells was also measured by bioluminescence in a stably transfected human breast cancer MCF-7 cell line with a reporter gene that allowed expression of the firefly luciferase enzyme under the control of an estrogen regulatory element.

Breast Neoplasms↗
PubMed2001

Direct analysis of semivolatile organic compounds in air by atmospheric pressure chemical ionization mass spectrometry.

Atmospheric pressure chemical ionization is employed for direct air analysis, without ion source modification, by using the sheath gas as the sample transport agent. A simple modification of the sheath gas inlet line allows introduction of gaseous samples into a commercial atmospheric pressure chemical ionization source. Optimization and testing of this novel air sampling method are described and detection of semivolatile compounds is shown. The analytical performance of the technique is established with methyl salicylate, including a limit of quantification of 100 pptr, a limit of detection of 50 pptr, a linear response from 100 pptr to 20 ppb, and rise and fall times of 12 and 20 s, respectively. Using reagent ion monitoring, it is shown that the protonated methanol dimer is the principal CI reagent ion leading to protonated dimethyl methylphosphonate, while the monomer is mainly responsible for protonating methyl salicylate. Since the formation of the CI reagent (methanol clusters) can be controlled by simple variation of experimental parameters, the selectivity of the method can be easily adjusted to suit the targeted analyte. Performance is found to be independent of the choice of air or nitrogen as the sheath gas (and thus as the sample matrix) and this, together with the sensitivity and speed of the technique, make it promising for field studies.

Journal Article↗
PubMed2001

External interface for trap-and-release membrane introduction mass spectrometry applied to the detection of inorganic chloramines and chlorobenzenes in water.

Construction and evaluation of an external configuration trap-and-release membrane introduction system for mass spectrometry is described. This novel interface allows independent control of the temperature of the membrane and eliminates the dependence of membrane heating efficiency on its position in the ion source. The external trap-and-release MIMS configuration is successfully applied to detection of inorganic chloramines and chlorobenzenes. The method is shown to give temporal resolution of volatile vs. semi-volatile compounds, which increases its sensitivity for semi-volatiles in the presence of volatiles and provides an additional selectivity parameter. Further selectivity is provided by tandem mass spectrometry.

Chloramines↗
PubMed2001

Effects of liquid phase composition on salt cluster formation in positive ion mode electrospray mass spectrometry: implications for clustering mechanism in electrospray.

Potassium bromate salt clusters, [KBrO3]nKx(x+), formed by electrospray ionization were studied as a function of solution properties. Clusters with up to 4 positive charges were observed. Their abundance, charge state and distribution were shown to vary with the organic solvent in solution. The effects of 7 solvents, including methanol, ethanol, isopropanol, acetonitrile, acetone, pyridine, and 1,4-dioxane, were thoroughly investigated. Solvents with a low dielectric constant and a high viscosity seem to favor clustering in solution but do not systematically allow high charge state ion formation. On the other hand, cluster charge reduction during desolvation was not correlated with solvent cation affinity over the range of solvents examined. However, ion distribution in mass spectra could be rationalized as a combination of these two competing phenomena. Charge state increases with the cluster size but may be reduced during ion desolvation when high cation affinity solvent molecules are actually involved in the ion solvation shell. This assumption could be envisaged in either Iribarne or Dole mechanisms of ion release in the gas phase. However, intensity profiles of multiply charged clusters could only be understood in terms of the ion evaporation mechanism.

Journal Article↗
PubMed2001

[Laparoscopic management of ovarian borderline tumors].

The objective of this communication is to present a patient case with ovarian borderline tumor. The conservative management for laparoscopy, and we performed an analysis of the mundial literature in which respect to the conservative and laparoscopy management, in spite of the laparoscopy management are of controversy.

Adult↗
PubMed1998

Analysis of oxyhalides in water by ion chromatography-ionspray mass spectrometry.

A sensitive method for analyzing chlorite, chlorate, bromate and iodate in water by ion chromatography (IC) coupled with ionspray tandem mass spectrometry (IS-MS-MS) has been developed. Prior to analysis, samples were subjected to off-line sample clean-up with Ba, Ag and H-form resins to remove sulfate, chloride and hydrogencarbonate, respectively. Oxyhalides in the purified samples were concentrated and separated on a short, high-performance anion-exchange column. An eluent consisting of ammonium nitrate in methanol-water (9:1, v/v) was found to be suitable for separating the analytes, while providing enhanced detector sensitivity. The coupling of IC with IS-MS-MS allows for the identification of the four oxyhalides mentioned above in a single run with very high specificity and sensitivity.

Drinking↗
PubMed1997

Choosing the right macrolide antibiotic. A guide to selection.

Macrolide antibiotics have proven to be valuable alternatives to penicillins and cephalosporins for the treatment of a number of infections. Currently, a number of macrolides are available. When choosing a particular macrolide, the types of organisms causing the infection, the tolerability of the drug, convenience of dosing and possible drug interactions all must be taken into account. Erythromycin, azithromycin and clarithromycin are equally effective against most gram-positive organisms. However, clarithromycin and azithromycin have much better activity against Haemophilus influenza and Moraxella catarrhalis. Thus, these 2 drugs are better choices for the treatment of community-acquired pneumonia. However, the low serum concentrations of azithromycin may be a problem in patients with bacteraemia associated with with community-acquired pneumonia. Clarithromycin appears to be effective for the treatment and prophylaxis of Mycobacterium avium complex (MAC) in patients with AIDS, while azithromycin appears to be effective for prophylaxis. Treatment of MAC with azithromycin is currently undergoing study. Although clarithromycin is the macrolide of choice for the treatment of Helicobacter pylori, azithromycin is the preferred macrolide for the treatment of Chlamydia trachomatis infections. The major factor limiting the use of azithromycin and clarithromycin has been their cost. However, these drugs may be cost effective if compliance is improved due to better tolerability and more convenient dosing regimens.

Acute Disease↗
PubMed1996

Laparoscopic Hysterectomy

Retrospective analysis of 200 laparoscopic hysterectomies performed from June 1991 to July 31, 1995, showed the main indications to be symptomatic uterine fibroids, advanced endometriosis, chronic pelvic inflammatory disease, and endometrial hyperplasia with atypia. Some women had more than one indication, and pathology reports contained more than one diagnosis. Three cases were converted to abdominal procedures. Operating room time was under 2 hours in general, and blood loss was around 120 ml. Most of the initial cases were performed with endostapling. This technique was changed in favor of bipolar and unipolar coagulation and suturing. Laparoscopic hysterectomy is a safe, superior technique that should be an integral part of gynecologic practice.

Journal Article↗
PubMed1996

Logical observation identifier names and codes (LOINC) database: a public use set of codes and names for electronic reporting of clinical laboratory test results.

Many laboratories use electronic message standards to transmit results to their clients. If all laboratories used the same "universal" set of test identifiers, electronic transmission of results would be greatly simplified. The Logical Observation Identifier Names and Codes (LOINC) database aims to be such a code system, covering at least 98% of the average laboratory's tests. The LOINC database should be of interest to hospitals, clinical laboratories, doctors' offices, state health departments, governmental healthcare providers, third-party payors, organizations involved in clinical trials, and quality assurance and utilization reviewers. The fifth release of the LOINC database, containing codes, names, and synonyms for approximately 6300 test observations, is now available on the Internet for public use. Here we describe the LOINC database, the methods used to produce it, and how it may be obtained.

Clinical Laboratory Information Systems↗
PubMed1994

Regulation of CTP: phosphocholine cytidylyltransferase in HepG2 cells: effect of choline depletion on phosphorylation, translocation and phosphatidylcholine levels.

We studied the effect of choline depletion on the biosynthesis of phosphatidylcholine (PC) and the distribution and phosphorylation of cytidylyltransferase (CT) in HepG2 cells. Phosphocholine concentrations decreased within 24 h of choline depletion to values less than 2% of controls. The incorporation of [3H]glycerol into PC was reduced in choline-depleted (CD) cells. The apparent turnover of PC was similar in CD and choline-supplemented (CS) cells (T1/2 = 20 h). The methylation pathway for PC synthesis increased nearly 10-fold in CD cells. Cell growth was similar in CD and CS cells. Over 95% of CT activity in CS cells was in the soluble pool. Choline depletion resulted in a progressive decrease in CT activity and immunodetected enzyme in the soluble pool and a corresponding increase in membrane CT over a 48-h period. Choline supplementation of CD cells caused a rapid release of membrane CT (complete release by 3 h). Two phosphorylated forms of CT were identified. One form contained a higher level of phosphorylation (HPCT) than the other form (LPCT). HPCT migrated slightly slower than LPCT on SDS gels. CD cells contained only LPCT in both soluble and membrane pools. CS cells contained only HPCT. During choline depletion PC content decreased nearly 20% but CT binding did not occur until LPCT was generated in cytosol. Conversely, choline supplementation released LPCT into cytosol and HPCT was formed only after the release. We conclude that both the induction of binding sites, perhaps by depletion of PC and dephosphorylation of HPCT to LPCT, are required for CT translocation to membranes. The release of CT from membranes is initiated by changes in membrane binding sites followed by trapping of the CT in the soluble pool by phosphorylation of LPCT to HPCT.

Binding Sites↗
PubMed1993

The occurrence of the transposable element pogo in Drosophila melanogaster.

We examined the genomic occurrence of the transposable element pogo in over 120 strains of Drosophila melanogaster, from around the world and from different eras. All had multiple copies of a 2.1 kilobase (kb) pogo element, and multiple copies of several size classes between 1.0 and 1.8 kb. There were differences between strains in intensities or presences of deletion-derivative size classes, suggesting current or recent mobility in the species. We were unable to find any pogo-hybridization in eight other species in the genus, in three subgenera, or in the related Scaptomyza pallida. The pogo element may be a 'middle-aged' element in the genome of D. melanogaster, having entered the species since its divergence from its sibling species, but long before the P and hobo elements.

Animals↗
PubMed1991

Control of phosphatidylcholine synthesis in Hep G2 cells. Effect of fatty acids on the activity and immunoreactive content of choline phosphate cytidylyltransferase.

We examined the effect of fatty acids on phosphatidylcholine synthesis and cytidylyltransferase activity in Hep G2 cells. Treatment of Hep G2 cells with oleic acid caused an increase in the incorporation of [methyl-14C]choline into phosphatidylcholine and a corresponding decrease in radioactivity in choline phosphate using a pulse-chase procedure. This result is consistent with a fatty acid-induced increase in the cytidylyl-transferase step in the choline pathway. We measured cytidylyltransferase activity in membrane fractions and in cytosol (100,000 x g supernatant or soluble enzyme released by digitonin). The activity increased in both membrane and cytosol. Thus, an increase in total activity occurred. Cytidylyltransferase protein determined by Western blot immunoassay increased after oleic acid treatment. Immunotitration of cytidylyltransferase protein also indicated that an increase in enzyme protein resulted from oleic acid treatment. Cycloheximide did not prevent the oleic acid-induced increase in cytidylyltransferase activity. The increase in enzyme activity was apparent when we measured the activity in the presence or absence of lipid activators. Separation of cytosolic cytidylyltransferase into H- and L-forms showed that the increase in cytosolic activity was due to an increase in H-form. The amount of L-form did not change. We interpret these results to suggest that fatty acid treatment of Hep G2 cells promoted the formation of active cytidylyltransferase (H-form) from a preexisting inactive form. The increased activity was distributed between membranes and the lipoprotein form in cytosol (H-form).

Blotting, Western↗
PubMed1990

CTP:phosphocholine cytidylyltransferase in rat lung: relationship between cytosolic and membrane forms.

The purpose of these studies was to determine the properties of the membrane-bound cytidylyltransferase in adult lung and to assess the relationship between the microsomal enzyme and the two forms of cytidylyltransferase in cytosol. Microsomes, isolated by glycerol density centrifugation, contained significantly less cytidylyltransferase than microsomes isolated by differential centrifugation (11.6 +/- 3.2 vs. 30 +/- 11 nmol/min per g lung). The released activity was recovered as H-form cytidylyltransferase. Cytidylyltransferase activity was not removed from microsomes by washing of the microsomal pellet with homogenizing buffer. Triton X 100 extracted all of the cytidylyltransferase from microsomes. The extracted activity was similar to H-form. Chlorpromazine dissociated microsomal enzyme to L-form. Chlorpromazine has been shown previously to dissociate H-form to L-form. These results suggested that microsomal cytidylyltransferase existed in a form similar if not identical to cytosolic H-form. In vitro translocation experiments demonstrated that the L-form of cytidylyltransferase was the species which binds to microsomal membranes. Triton X 100 extraction of microsomes from translocations experiments removed the bound enzyme activity. Glycerol density fractionation indicated that the activity in the Triton extract was H-form cytidylyltransferase. We concluded that the active lipoprotein form of cytidylyltransferase (H-form) is the membrane-associated form of cytidylyltransferase in adult lung; that it is formed after the L-form binds to microsomal membranes and that cytosolic H-form is released from the membrane.

Animals↗
PubMed1989

Characterization of cytosolic forms of CTP: choline-phosphate cytidylyltransferase in lung, isolated alveolar type II cells, A549 cell and Hep G2 cells.

The subcellular forms of cytidylyltransferase (EC 2.7.7.15) in rat lung, rat liver, Hep G2 cells, A549 cells and alveolar Type II cells from adult rats were separated by glycerol density centrifugation. Cytosol prepared from lung, Hep G2 cells, A549 cells and alveolar Type II cells contained two forms of the enzyme. These species were identical to the L-Form and H-Form isolated previously from lung cytosol by gel filtration. Liver cytosol contained only the L-Form. Rapid treatment of Hep G2 cells with digitonin released all of the cytoplasmic cytidylyltransferase activity. The released activity was present in both H-Form and L-Form. The molecular weight of L-Form was determined from sedimentation coefficients and Stokes radius values to be 97,690 +/- 10,175. Thus, the L-Form appears to be a dimer of the Mr 45,000 catalytic subunit. The f/f degrees value of 1.5 indicated that the protein molecule has an axial ratio of 10, assuming a prolate ellipsoid shape. The estimated molecular weight of the H-Form was 284,000 +/- 25,000. The H-Form was dissociated into L-Form by incubation of cytosol at 37 degrees C. Triton X-100 (0.1%) and chlorpromazine (1.0 mM) also dissociated the H-Form into L-Form. Western blot analysis indicated that both forms contained the catalytic subunit. An increase in Mr 45,000 subunit coincided with the increase in cytidylyltransferase activity in L-Form, which resulted from the dissociated of H-Form. The L-Form was dependent on phospholipid for activity. The H-Form was active without lipid. Phosphatidylinositol was present in the H-Form isolated from Hep G2 cells. The phosphatidylinositol dispersed when the H-Form was dissociated into L-Form. Phosphatidylinositol and phosphatidylglycerol cause L-Form to aggregate into a form similar to H-Form. Phosphatidylcholine/oleic acid (1:1 molar ratio) and oleic acid also aggregated the L-Form. Phosphatidylcholine did not produce aggregation. We conclude that the H-Form is the active form of cytidylyltransferase in cytoplasm. The H-Form appears to be a lipoprotein consisting of an apoprotein (L-Form dimer of the Mr 45,000 subunit) complexed with lipids. A change in the relative distribution of H-Form and L-Form in cytosol would alter the cellular activity and thus may be important in the regulation of phosphatidylcholine synthesis.

Adenoma↗
PubMed1987

Role of a guanine nucleotide regulatory protein in the activation of phospholipase C by different chemoattractants.

It is well established that formyl peptide chemoattractants can activate a phospholipase C in leukocytes via a pertussis toxin (PT)-sensitive guanine nucleotide regulatory (G) protein. Whether this pathway is similarly used by chemoattractant receptors as a class has been unclear. We now report that lipid and peptide chemoattractants in direct comparative studies induced similar amounts of initial (less than or equal to 15 sec) inositol trisphosphate (IP3) release in human polymorphonuclear leukocytes, but the response to lipid chemoattractants was more transient. Production of IP3 by all chemotactic factors was inhibited by treatment of the cells with PT, indicating that chemotactic factor receptors as a class are coupled to phospholipase C via a G protein that is a substrate for ADP ribosylation by PT. The peptide and lipid factors had comparable chemotactic activity, which was also inhibitable by PT. However, transient activation of phospholipase C is apparently an insufficient signal for full cellular activation, since the lipid chemotactic factor leukotriene B4 and platelet-activating factor were poor stimuli for O2- production and lysosomal enzyme secretion compared with N-formyl-methionyl-leucyl-phenylalanine (fMet-Leu-Phe). Nonetheless, treatment with PT inhibited O2- production and enzyme secretion in response to all chemoattractants, but as previously noted, did not affect Ca2+ ionophores, lectins, or phorbol myristate acetate. Formyl peptide and lipid chemotactic factors induced similar levels of Ca2+ mobilization when monitored by Quin 2 or chlortetracycline (CTC) fluorescence. Although these responses to fMet-Leu-Phe were blocked by PT, the Quin 2 and initial CTC response to the lipid factors were only partially susceptible. Thus, the lipid factors apparently utilize an additional PT-resistant mechanism for redistributing intracellular Ca2+. This latter process requires extracellular Ca2+ and may be independent of the PT-sensitive G protein.

Calcium↗
PubMed1981

A four-year follow-up study of the effects of methylphenidate on the behavior and academic achievement of hyperactive children.

Sixty-two children were evaluated 4 years after their initial referral for symptoms of hyperactivity. Behavioral measures included parent and teacher judgments of behavior and social adjustment. Academic achievement was assessed by teachers' reports, number of failed grades, special education services, and two individually administered achievement tests. Data were analyzed for the total group to determine both the extent to which presenting problems diminished over time and the major problems still present. The effects of stimulant drug therapy on outcome were assessed by dividing the children into groups according to the duration of time they had taken stimulants. Total duration of time on stimulants ranged from less than 6 months (group 1) to 4 years (group 5). Results indicated that the symptoms of hyperactivity significantly lessened, but remained higher in these children than in normal peers. Behavioral and social problems were less pervasive than academic underachievement. There were no group differences, indicating that the duration of stimulant intervention did not have a significant effect on outcome. The clinical implication of this study is that the beneficial effects of stimulant drug intervention occur within the first months after initiation of therapy. Long-term treatment does not appear to be of value in producing better outcome.

Achievement↗