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M Charlton

Publications and source records attributed to M Charlton.

At least 19 recordsLinked to original sources
PubMed2001

Insulin's effect on synthesis rates of liver proteins. A swine model comparing various precursors of protein synthesis.

Insulin's effect on the synthesis of liver proteins remains to be fully defined. Previous studies using various surrogate measures of amino acyl-tRNA have reported variable results of insulin's effect on liver protein synthesis. We determined the effect of insulin with or without amino acid supplementation on the synthesis rates of liver proteins (tissue, albumin, and fibrinogen) using L-[1-13C]Leu as a tracer in 24 male miniature swine. In addition, we compared the isotopic enrichment of different precursors of liver proteins with that of amino acyl-tRNA using L-[1-13C]Leu and L-[15N]Phe as tracers. Although liver tissue fluid enrichment of [13C]Leu and [15N]Phe and that of plasma [13C]ketoisocaproatic acid (KIC) were very similar to that of tRNA, plasma isotopic enrichment of both Leu and Phe were substantially higher (P < 0.01) and VLDL apolipoprotein-B100 enrichment was lower (P < 0.01) than the respective amino acyl-tRNA enrichment. Plasma KIC enrichment most accurately predicted leucyl-tRNA enrichment, whereas plasma Leu enrichment was best correlated with that of tRNA. Neither insulin alone nor insulin plus amino acid infusion had an effect on liver tissue protein synthesis. In contrast, insulin alone decreased the albumin synthesis rate, and insulin with amino acids maintained the albumin synthesis rate. Insulin with or without amino acids inhibited the fibrinogen synthesis rate. These results, based on synthetic rates using amino acyl-tRNA, were consistent with those obtained using KIC or tissue fluid Leu or Phe as precursor pools. These studies demonstrated that plasma KIC enrichment is a convenient and reliable surrogate measure of leucyl-tRNA in liver. We also concluded that insulin has differential effects on the synthesis rates of liver proteins. Whereas insulin with or without amino acid supplement has no acute effect on the synthesis of liver tissue protein, insulin has a substantial inhibitory effect on fibrinogen synthesis. In contrast, insulin administration along with amino supplement is necessary to maintain albumin synthesis rate.

Amino Acids↗
PubMed2000

Impact of nutritional status on outcomes after liver transplantation.

BACKGROUND: Poor preoperative nutritional status has been reported to be associated with adverse outcomes after liver transplantation. Published data are, however, conflicting, with methods of preoperative nutritional assessment and postoperative outcomes varying between studies. METHODS: We prospectively studied the predictive value of preoperative nutritional status for adverse outcomes after liver transplantation. Assessment of preoperative nutritional status included: body cell mass determination, subjective global assessment, anthropometry, handgrip dynamometry, biochemical and amino acid profile, Child's score, and dual-energy x-ray absorptiometry. Death, intensive care unit (ICU) length of stay > or =4 days, hospital length of stay > or =15 days, blood usage > or =36 U of blood products, infection, rejection, and global resource utilization (an index of cost) greater than the median were considered poor outcomes. RESULTS: Fifty-three patients were studied. Longer ICU stay was associated with lower handgrip strength (P<0.01) and lower aromatic amino acid levels (P<0.01). Longer total hospital stay and the development of infections were associated with lower branched chain amino acid levels (P<0.01 and <0.001, respectively). Acute cellular rejection was associated with lower total body fat (P<0.001) and higher triglyceride levels (P<0.02). Neither death nor higher global resource utilization was associated with any preoperative nutritional parameter. CONCLUSIONS: Lower preoperative handgrip strength and branched chain amino acid levels are associated with longer ICU stays and increased likelihood of posttransplant infections. In our program, in which nutritional support was provided to potential recipients exhibiting malnourishment, none of the measured nutritional parameters were associated with mortality or greater global resource utilization.

Adult↗
PubMed2000

The effect of insulin on human small intestinal mucosal protein synthesis.

BACKGROUND & AIMS: Insulin deficiency was recently shown to stimulate splanchnic protein synthesis in vivo, whereas insulin enhances small intestinal mucosal cell proliferation in vitro. Because insulin is a postprandial hormone, it was hypothesized that it has an important role in regulating small intestinal protein synthesis in humans. METHODS: Small intestinal mucosal protein synthesis was measured in C-peptide-negative patients with type 1 diabetes mellitus during insulin deprivation (n = 6) and during insulin treatment (n = 6) and in nondiabetic control subjects (n = 6). Mucosal protein synthesis was measured from the increment of [(13)C]leucine enrichment in endoscopically obtained duodenal mucosa samples during a primed continuous infusion of L-[1-(13)C]leucine. RESULTS: During insulin treatment, the rate of mucosal protein synthesis in patients with type 1 diabetes was similar (1.32% +/- 0.05%/h) to that of nondiabetic controls (1.33% +/- 0.06%/h). However, during insulin deprivation, the mucosal protein synthesis rate in patients with type 1 diabetes was significantly lower (1.15% +/- 0.33%/h) than during either insulin treatment (P = 0.01) or in nondiabetic controls (P = 0.04). CONCLUSIONS: These studies show that insulin is required for the maintenance of normal rates of protein synthesis in small intestinal mucosa. Because protein synthesis is an essential component of the remodeling process of this fast turning over tissue, the decline in the synthesis rate of small intestinal mucosa during insulin deprivation may be a contributing factor in the development of gastrointestinal complications that occur in poorly controlled type 1 diabetic patients.

Adult↗
PubMed2000

A prospective analysis of 1,930 patients with hematuria to evaluate current diagnostic practice.

PURPOSE: The commonly accepted diagnostic algorithm for hematuria includes excretory urography (IVP) and cystoscopy. Some have suggested that ultrasound of the upper urinary tract is adequate and that cystoscopy is not necessary in younger patients with microscopic hematuria. We ascertain whether a less intensive algorithm could be adopted while retaining diagnostic efficacy. MATERIALS AND METHODS: A total of 1,930 patients were enrolled prospectively in the study at a hematuria clinic between October 1994 and March 1997. Evaluation consisted of basic demographics, history and examination, routine blood tests, urinalysis and cytology. All patients underwent plain abdominal radiography, renal ultrasound, IVP and flexible cystoscopy. RESULTS: A total of 1,194 males and 736 females with a mean age of 58 years (range 17 to 96) were included in the study. Overall, 61% of patients had no basis found for hematuria, 12% had bladder cancer, 13% had urinary tract infection and 2% had stones. Kidney and upper tract tumors were noted in 14 patients (0.7%), including 4 who presented with microscopic hematuria. If only ultrasound or IVP had been performed 4 of these cases would have been missed. Of 982 patients presenting with microscopic hematuria 51 had cancer. Bladder cancer was found in 7 patients younger than 40 years. CONCLUSIONS: Our findings suggest that cystoscopy cannot be safely avoided even in younger patients with microscopic hematuria. Only a combination of ultrasound and IVP detected all upper tract tumors.

Adolescent↗
PubMed1999

Impact of immunosuppression and acute rejection on recurrence of hepatitis C: results of the National Institute of Diabetes and Digestive and Kidney Diseases Liver Transplantation Database.

Whereas the impact of early (first 6 postoperative weeks) acute cellular rejection on patient survival among liver transplant recipients as a whole has been reported to be favorable, we hypothesized treatment for acute cellular rejection may have differing impacts on patient and graft survival in hepatitis C virus (HCV)-infected and HCV-negative transplant recipients. We studied the impact of immunosuppression and rejection on patient and graft survival among the 166 HCV-infected and 602 HCV-negative transplant recipients enrolled onto the National Institute of Diabetes and Digestive and Kidney Diseases Liver Transplantation Database. All data were collected prospectively. The association of early acute cellular rejection with mortality was determined using a Cox proportional hazards model with a time-dependent covariate. Median follow-up was 5.0 years for HCV-infected and 5.2 years for HCV-negative transplant recipients. HCV-infected transplant recipients experienced similar frequencies of acute cellular and steroid-resistant rejection as patients undergoing liver transplantation for most other indications. The mortality risk was significantly increased (relative risk = 2.4; P =.03) for HCV-infected transplant recipients who developed early acute cellular rejection compared with HCV-negative transplant recipients. None of the HCV-infected transplant recipients developed allograft failure secondary to chronic rejection. The choice of calcineurin inhibitor did not affect posttransplantation outcomes. Early acute cellular rejection occurs at similar frequencies in HCV-infected and HCV-negative transplant recipients. Although an episode of early acute cellular rejection is associated with a lower cumulative mortality among HCV-negative transplant recipients, the opposite is true for HCV-infected transplant recipients, who experience an increased risk for mortality after an episode of early acute cellular rejection. The adverse impact of early acute cellular rejection on patient survival should be considered in developing primary immunosuppression and acute cellular rejection treatment protocols for HCV-infected transplant recipients.

Databases, Factual↗
PubMed1998

Predictors of patient and graft survival following liver transplantation for hepatitis C.

End-stage liver disease secondary to hepatitis C virus (HCV) infection is the leading indication for liver transplantation in the United States. Recurrence of HCV infection is nearly universal. We studied the patients enrolled in the National Institute of Diabetes and Digestive and Kidney Diseases Liver Transplantation Database to determine whether pretransplantation patient or donor variables could identify a subset of HCV-infected recipients with poor patient survival. Between April 15, 1990, and June 30, 1994, 166 HCV-infected and 509 HCV-negative patients underwent liver transplantation at the participating institutions. Median follow-up was 5.0 years for HCV-infected and 5.2 years for HCV-negative recipients. Pretransplantation donor and recipient characteristics, and patient and graft survival, were prospectively collected and compared. Cumulative patient survival for HCV-infected recipients was similar to that of recipients transplanted for chronic non-B-C hepatitis, or alcoholic and metabolic liver disease, better than that of patients transplanted for malignancy or hepatitis B (P = .02 and P = .003, respectively), and significantly worse than that of patients transplanted for cholestatic liver disease (P = .001). Recipients who had a pretransplantation HCV-RNA titer of > or = 1 x 10(6) vEq/mL had a cumulative 5-year survival of 57% versus 84% for those with HCV-RNA titers of < 1 x 10(6) vEq/mL (P = .0001). Patient and graft survival did not vary with recipient gender, HCV genotype, or induction immunosuppression regimen among the HCV-infected recipients. While long-term patient and graft survival following liver transplantation for end-stage liver disease secondary to HCV are generally comparable with that of most other indications, higher pretransplantation HCV-RNA titers are strongly associated with poor survival among HCV-infected recipients.

Adult↗
PubMed1998

TT-virus infection in North American blood donors, patients with fulminant hepatic failure, and cryptogenic cirrhosis.

A novel DNA virus, TT-virus (TTV), has been reported in patients with non-A-G posttransfusion hepatitis in Japan. We sought to determine whether TTV infection occurs in North American blood donors and to further determine the prevalence of TTV infection in several groups of patients with liver disease, including patients with cryptogenic cirrhosis and idiopathic fulminant hepatic failure. TTV infection was sought by detection of TTV DNA in serum by polymerase chain reaction (PCR) using primers generated from a conserved region of the TTV genome. Blood donors, patients with cryptogenic cirrhosis, idiopathic fulminant hepatic failure, and patients with other forms of advanced liver disease with and without a history of parenteral exposures were studied. TTV infection was present in 1% (1 of 100) of blood donors, 15% (5 of 33) of patients with cryptogenic cirrhosis, 27% (3 of 11) of patients with idiopathic fulminant hepatic failure, 18% (2 of 11) of patients with a history of exposure to blood products, and 4% (1 of 25) of patients without parenteral risk factors. For all patients tested, a history of prior exposure to blood products was associated with an increased risk of TTV infection (relative risk, 4.5; 90% confidence intervals, 0.6-43.9). We conclude that TTV infection is present among North American blood donors and is common in patients with liver disease, including cryptogenic cirrhosis and fulminant hepatic failure. Further studies are required to determine the role of TTV in the pathogenicity of acute and/or chronic liver disease.

Adult↗
PubMed1998

Pyrithione biocide interactions with bacterial phospholipid head groups.

Sodium pyrithione and zinc pyrithione (NaPT and ZnPT, respectively) are antimicrobial agents widely used in both the cosmetics and fuel industries. They are also utilized in the mining industry because of their metal chelating properties. They have been shown to depolarize membrane electropotential in fungi and are also known to inhibit fungal and bacterial substrate transport processes. Recent work has shown that both pyrithiones cause the leakage of intracellular material (potassium ions and O.D.260 nm absorbing material) from exposed bacterial cells. The work here reports studies on the interactions between the pyrithiones and the bacterial phospholipid head group structures, at both a practical and a theoretical level, utilizing tube dilution neutralizer studies, scanning spectrophotometry and molecular modelling. The tube dilution neutralizer studies exhibited a decrease in minimum inhibitory concentration (MIC) for both pyrithiones in the presence of extracellular phosphatidyl-ethanolamine and EDTA. Scanning spectrophotometry exhibited the chelation of the central zinc atom from the ZnPT chelate by the addition of EDTA. Molecular modelling studies exhibited the chelation of the phosphatidyl-ethanolamine head group by ZnPT. Zinc pyrithione also exhibited an interaction with the ammonium tail of the head group structures. Sodium pyrithione exhibited electrostatic interactions with the phospholipid head groups in the molecular modelling studies.

Antifungal Agents↗
PubMed1998

Protein metabolism in insulin-dependent diabetes mellitus.

Patients with insulin-dependent diabetes are in a catabolic state without insulin replacement. The mechanism of insulin's anticatabolic effect has been investigated in whole-body and regional tracer kinetic studies. Whole-body studies have demonstrated that there are increases in both protein breakdown and protein synthesis during insulin deprivation. Because the magnitude of the increase in protein breakdown is greater than the magnitude of the increase in protein synthesis, there is a net protein loss during insulin deprivation. Regional studies have shown that insulin replacement inhibits protein breakdown and synthesis in splanchnic tissue but only inhibits protein breakdown in skeletal muscle. Because the increase in protein synthesis in splanchnic tissues is greater than the increase in protein breakdown, insulin deprivation results in a net accretion of protein in the splanchnic bed. In contrast, in skeletal muscle, there is a net increase in protein breakdown during insulin deprivation, resulting in a net release of amino acids. There are no human data concerning the site of protein accretion in the splanchnic bed or the specific protein whose synthesis is increased during insulin deprivation. It appears that insulin exerts its overall anticatabolic effect in insulin-dependent diabetes mainly through the inhibition of muscle protein breakdown.

Diabetes Mellitus, Type 1↗
PubMed1995

Biochemical actions of chronic ethanol exposure in the mesolimbic dopamine system.

In previous studies, we have demonstrated that chronic administration of morphine or cocaine produces some common biochemical adaptations in the ventral tegmental area (VTA) and nucleus accumbens (NAc), components of the mesolimbic dopamine system implicated in the reinforcing actions of these and other drugs of abuse. Since this neural pathway is also implicated in the reinforcing actions of ethanol, it was of interest to determine whether chronic ethanol exposure results in similar biochemical adaptations. Indeed, as seen for chronic morphine and cocaine treatments, we show here that chronic ethanol treatment increased levels of tyrosine hydroxylase and glial fibrillary acidic protein immunoreactivity, and decreases levels of neurofilament protein immunoreactivity, in the VTA. Also like morphine and cocaine, ethanol increases levels of cyclic AMP-dependent protein kinase activity in the NAc. These actions of ethanol required long-term exposure to the drug, and were in most cases not seen in the substantia nigra or caudateputamen, components of the nigrostriatal dopamine system studied for comparison. Altered levels of tyrosine hydroxylase in catecholaminergic cells frequently reflect altered states of activation of the cells. Moreover, increasing evidence indicates that ethanol produces many of its acute effects on the brain by regulating NMDA glutamate and GABAA receptors. We therefore examined the influence of chronic ethanol treatment on levels of expression of specific glutamate and GABA receptor subunits in the VTA. It was found that long-term, but not short-term, ethanol exposure increased levels of immunoreactivity of the NMDAR1 subunit, an obligatory component of NMDA glutamate receptors, and of the GluR1 subunit, a component of many AMPA glutamate receptors; but at the same time, long-term ethanol exposure decreased immunoreactivity levels of the alpha 1 subunit of the GABAA receptor complex. These changes are consistent with an increased state of activation of VTA neurons inferred from the observed increase in tyrosine hydroxylase (TH) expression. These results demonstrate that chronic ethanol exposure results in several biochemical adaptations in the mesolimbic dopamine system, which may underlie prominent changes in the structural and functional properties of this neural pathway related to alcohol abuse and alcoholism.

Adenylyl Cyclases↗
PubMed1995

Potentiation of a slow Ca(2+)-dependent K+ current by intracellular Ca2+ chelators in hippocampal CA1 neurons of rat brain slices.

1. In hippocampal CA1 neurons of rat brain slices, a Ca(2+)-dependent slow afterhyperpolarization (sAHP) and underlying K+ current (IsAHP) are activated by Ca2+ influx and presumably reflect the time course of the intracellular Ca2+ signal produced by neuronal stimulation. We tested the hypothesis that when exogenous Ca2+ chelators become the predominant mobile Ca2+ buffer in the neuron, they alter the shape of intracellular Ca2+ signals responsible for IsAHP. The nature of this alteration provides insight into the mechanism of IsAHP generation. 2. Derivatives of 1,2-bis-[2-amino phenoxy] ethane N,N,N',N' tetraacetic acid (BAPTA) with different dissociation constants (KDS) for Ca2+ ranging from 0.15 to 7,000 microM were used to test this hypothesis. We also examined the effects of ethylene glycolbis (beta-aminoethyl either)-N,N,N',N'-tetraacetic acid (EGTA), which has a KD similar to that of BAPTA for Ca2+, but which binds and releases Ca2+ 100 times more slowly. When delivered to the cytoplasm by dialysis from a patch pipette, these chelators potentiated, inhibited, or had no effect on IsAHP depending on their concentration, affinity, and binding kinetics. 3. IsAHP decayed exponentially over much of its time course, with a half-decay time of 0.9 +/- 0.1 s (mean +/- SE, n = 22). Immediately after breakthrough into the whole cell configuration, there was an initial period of approximately 5 min during which IsAHP magnitude increased approximately 3.5-fold with no change in time course. Thereafter, the time course and amplitude of IsAHP were stable for > 45 min. 4. Addition of 1 mM of the high-affinity chelators 5,5'-dimethyl BAPTA or BAPTA to the pipette solution first increased the decay time of IsAHP 1.5-fold. However, within 10-15 min after break-through, the current was abolished. Addition of Ca2+ (0.1-1.0 mM) to the patch pipette containing the BAPTA derivatives reduced the ability of a given concentration of high-affinity chelator to inhibit IsAHP and also prolonged the period of IsAHP enhancement. A similar prolongation of the period of enhancement with even less attenuation of IsAHP was apparent with 0.1 mM 5,5'-dimethyl BAPTA and 0.1 mM Ca2+. 5. The intermediate-affinity chelator 4.4'-difluoro BAPTA (1 mM) prolonged the decay phase of the sAHP/IsAHP without attenuating the current. A twofold prolongation of IsAHP also was observed in neurons dialyzed with internal solution containing 3 mM EGTA and 0.3 mM Ca2+. Dialysis with 1 mM of the low-affinity chelators 2-amino-5-fluorophenol-N,N,O-triacetic acid (5-fluoro APTRA) or 5,5'-dinitro BAPTA had no apparent effect on IsAHP. All of the chelators that prolonged the decay phase of IsAHP also induced a rising phase such that a well-defined peak of IsAHP could be discerned at approximately 0.6 s after the end of the stimulus used to evoke the current. 6. Weak stimulation of muscarinic receptors selectively inhibits IsAHP. Thus the uncontaminated time course of IsAHP can be deduced by subtracting currents recorded before and after such muscarinic stimulation. With minimal exogenous buffer in the pipette (0.1 mM EGTA), the muscarinic-receptor-sensitive current exhibited a rising phase lasting approximately 300 ms and then decayed with a half-time of approximately 1 s. Both the rising and decay phases of the muscarinic-receptor-sensitive current were prolonged at least twofold by dialysis with BAPTA or 4,4'-difluoro BAPTA. Thus the effect of the chelators on the time course of IsAHP is not simply and artifact of inhibition of early components of the outward current. 7. The effects of BAPTA analogues on the time course of IsAHP are not due to changes in mobilization of intracellular Ca2+. External application of caffeine (10 mM), ryanodine (20 microM), dantrolene (20 microM), or thapsigargin (100 microM) had no effect on IsAHP recorded with the standard pipette solution or

Adaptation, Physiological↗