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Biomedical subjects

L J Langman

Publications and source records attributed to L J Langman.

18 recordsLinked to original sources
PubMed2003

Persistence of vitamin B12 insufficiency among elderly women after folic acid food fortification.

OBJECTIVE: To estimate the associated risk of folate and vitamin B12 (B12) insufficiency, as well as vitamin repletion, following folic acid food fortification. DESIGN: Retrospective cross-sectional study over a 5-year period. SETTING: Two large laboratory databases in the provinces of Ontario and British Columbia, Canada. PARTICIPANTS: Canadian women aged 65 years and over who underwent concomitant clinical testing of serum folate and B12 during the pre-fortification period of January 1996 to December 1997 in Ontario (n = 733) and British Columbia (n = 3839), and in the near-complete post-fortification period of January 1998 to December 2000 in Ontario (n = 4415) and British Columbia (n = 6677). MEASUREMENTS: Geometric mean concentrations of serum folate and B12 before and after folate fortification. Prevalence ratios (PR) were used to separately compare the post- and pre-fortification period rates of folate deficiency (below 6.0 nmol/L); B12 insufficiency (below 150 pmol/L); and B12 insufficiency in combination with supraphysiological concentrations of serum folate (above 45 nmol/L). RESULTS: The mean baseline folate and B12 concentrations were similar between provinces. Using the combined provincial data, the mean serum folate concentration increased by 64% after fortification, from 14.8 to 24.2 nmol/L (p < 0.001). The average B12 concentration increased from 280 to 300 pmol/L, which was more pronounced in BC (p < 0.001) than in Ontario (p = 0.16). The prevalence of folate deficiency declined from 6.3% to 0.88% after fortification (PR 0.14, 95% confidence interval [CI] 0.11-0.18), while the decline in B12 deficiency was less pronounced (PR 0.78, 95% CI 0.71-0.86). CONCLUSIONS: The prevalence of combined B12 insufficiency with supraphysiological concentrations of serum folate increased from 0.09% pre-fortification to 0.61% post (PR 7.0, 95% CI 2.6-19.2). The introduction of folic acid food fortification was associated with a substantial improvement in the folate status of Canadian women aged 65 years and older, paralleled by a large decline in the rate of folate deficiency. Improvement in the B12 status of these women was far less pronounced. Because the prevalence of combined B12 insufficiency and supraphysiological concentrations of serum folate may have increased with folic acid food fortification, consideration should be given to confirming this finding, and possibly, to the addition of B12 to folate fortified foods.

Aged↗
PubMed2000

Hyperhomocyst(e)inemia and the increased risk of venous thromboembolism: more evidence from a case-control study.

BACKGROUND: Elevation of plasma homocyst(e)ine level is an independent risk factor for arterial and venous thrombosis. We studied the degree to which hyperhomocyst(e)inemia contributes to the development of venous thromboembolism, using a retrospective case-control study design. METHODS: Cases were individuals with objectively confirmed venous thromboembolism and no history of atherosclerosis seen at the Toronto Hospital Thrombosis Clinic, Toronto, Ontario, between January 1, 1996, and July 31, 1998. Three controls were matched for every case according to sex and age within 5 years and were derived from a large community cohort. All subjects underwent assessment for fasting plasma homocyst(e)ine levels. Hyperhomocyst(e)inemia was defined as a fasting total homocyst(e)ine concentration above the 95th percentile control value. RESULTS: Seventy cases and 210 matched controls were included. Men and women were equally represented, and most were younger than 60 years. Among cases with venous thromboembolism, the mean (+/- SD) plasma homocyst(e)ine level was significantly higher than in controls (13.0 +/- 6.9 micromol/L vs 9.0 +/- 4.8 micromol/L, respectively; P<.001). Sixteen (23%) of 70 cases had hyperhomocyst(e)inemia compared with 10 (5%) of 210 controls (odds ratio, 5.9; 95% confidence interval [CI], 2.5-13.8). Among subjects aged 60 years or younger, the odds ratio was 4.9 (95% CI, 1.4-16.4), while for those aged 60 years or older, it was 7.3 (95% CI, 2.2-24.0). Even with the exclusion of cases showing abnormal renal function or low serum vitamin B12 or folate levels, the odds ratio remained significantly elevated at 3.3 (95% CI, 1.1-10.0). CONCLUSIONS: We found that fasting hyperhomocyst(e)inemia is a significant risk factor for venous thromboembolic disease in patients at a thrombosis clinic. Given the magnitude of effect and consistency across these studies, it is likely that homocyst(e)ine plays a causative role in the development of venous thrombosis, and it should be considered in the workup for venous thromboembolism.

Adult↗
PubMed1999

Homocysteine: cholesterol of the 90s?

Homocysteine is a sulfur-containing amino acid generated through the demethylation of methionine. It is largely catabolized by trans-sulfuration to cysteine but it may also be remethylated to methionine. Dubbed 'the cholesterol of the 90s' by the lay press, homocysteine is thought to be thrombophilic and to damage the vascular endothelium. Total plasma homocysteine (tHcy) is now established as a clinical risk factor for coronary artery disease, as well as other arterial and venous occlusive disease in adult populations. Regulation of homocysteine is dependent on nutrient intake, especially folate, vitamins B6 and B12. It is also controlled by common genetic variations (polymorphisms) in how vitamins are utilized as cofactors in the reactions controlling homocysteine metabolism. Moreover, concentrations are age- and sex-dependent and are altered by renal function, hormonal status, drug intake and a variety of other common clinical factors. Considerable care must be taken in assaying tHcy. Plasma should be separated shortly after collection, to avoid artifactual increases due to synthesis by blood cells in vitro. Reference methods have not been validated and criteria for establishing reference ranges should take into account the variable prevalence of physiological hyperhomocysteinemia. Determination of tHcy should probably be limited to centres with relevant expertise and ability to maintain the high degree of precision required for reliable interpretation. Molecular testing for the genetic polymorphisms is still in the research phase but the ease and reliability of molecular diagnosis will speed its introduction into clinical laboratory practice--particularly in relation to diagnosis of thrombophilic disorders. Clinical research initiatives are being driven by the benefit that should be achieved by correction with vitamin supplements, particularly folate and B vitamins, but it must be recognized that prospective controlled studies to validate clinical benefit are only now being initiated. At the moment, it is safe to say that hyperhomocysteinemia is one of the few prevalent biochemical risk factors for thromboembolic disease that might be corrected by vitamin supplements. Such a possibility lies behind the growing momentum to recommend increased supplements of folate and B vitamins to at-risk population and patient groups today.

Adult↗
PubMed1999

Homocysteine.

Homocysteine is a sulfur-containing amino acid generated through the demethylation of methionine. It is largely catabolized by trans-sulfuration to cysteine, but it may also be remethylated to methionine. Regulation of homocysteine is dependent on nutrient intake, especially folate, vitamins B6 and B12. It is also controlled by individual genetic differences in how vitamins are utilized as cofactors in the reactions controlling homocysteine metabolism. In excess quantities, homocysteine is thought to be thrombophilic and to damage the vascular endothelium. Total plasma homocysteine (tHcy) is now established as a clinical risk factor for coronary artery disease, as well as other arterial and venous occlusive disease in adult populations. These effects are probably related to its role as a teratogen in the pathogenesis of neural tube defects--genetic variants causing hyperhomocysteinemia are associated with both neural tube defects in susceptible pregnancies and with risks for vaso-occlusive disease in later years. Considerable care must be taken in assaying tHcy. Plasma should be separated shortly after collection to avoid artifactual increases due to synthesis by blood cells in vitro. tHcy concentrations must be interpreted in light of the fact that serum albumin, urate, creatinine, and vitamin concentrations may be important analytical covariates. Moreover, concentrations are age- and sex-dependent and are altered by renal function, hormonal status, drug intake, and a variety of other common clinical factors. Why then is homocysteine now of such great clinical and scientific interest? If the homocysteine moiety itself is important in the pathogenesis of vaso-occlusive disease, then simple treatment of hyperhomocysteinemia with vitamins should lead to a significant reduction in disease risk. Such a possibility lies behind the growing momentum to recommend increased supplements of folate and B vitamins to at-risk populations and patient groups today.

Amino Acid Metabolism, Inborn Errors↗
PubMed1998

Interference of IgM paraproteins in the Olympus AU800 uric acid assay.

INTRODUCTION: In the Olympus uric acid procedure, uric acid is converted by uricase to allantoin and hydrogen peroxide, which is reacted in a Trinder reaction to produce a chromophore read bichromatically at 520 and 660 nm. Repeated difficulty was encountered in obtaining uric acid results on samples from myeloma patients with known IgM paraproteins. Large absorbances in sample blanks were due to a visible precipitation observed in the reaction cuvettes. OBJECTIVE: To alter the Olympus method (OM) to eliminate the interference by IgM, and to verify the modified method (MM). METHODS: Dilution of the sample blank by saline was substituted for water in the MM, with small alterations in the reaction timing sequence necessary to accommodate the instrument requirements. RESULTS: A comparison of uric acid results obtained from nonmyeloma patient samples using the OM and the MM showed a good correlation (r = 0.970), and no statistical difference between the two means using a paired t-test. A similar comparison performed using the samples containing IgA and IgG paraproteins also revealed a good correlation (r = 0.981), and no statistical difference between the two means. Results on IgM containing specimens were assessed indirectly because the samples could not be assayed with the OM. First, removal of detectable levels of proteins using a 20% TCA solution did not affect the measurement of uric acid. Second, protein-free supernatants from IgM containing samples were measured by the OM and compared with the corresponding serum samples measured by the MM. There was good correlation between the two methods (r = 0.945), and no statistical difference between the means using a paired t-test. CONCLUSION: The modified method is satisfactory for routine analysis of samples, including those with IgM paraproteins.

Blood Chemical Analysis↗
PubMed1998

Correlation between total homocysteine and cyclosporine concentrations in cardiac transplant recipients.

Increased circulating total homocysteine (tHcy) has been implicated as an independent risk factor for atherosclerotic disease. In cardiac transplant patients, accelerated coronary atherosclerosis is an important cause of late allograft failure; however, studies of tHcy in this at-risk group are limited. We sampled a cohort of 72 subjects 3.95+/-3.14 (mean +/- SD) years after transplantation and found that all had tHcy concentrations above our upper reference limit (15.0 micromol/L). The mean tHcy in the transplant group (25.4+/-7.1 micromol/L) was significantly greater than in our reference group (9.0+/-4.3 micromol/L; n = 457; P <0.001). We also examined the effect of age, gender, time since transplant, serum folate and cobalamin, total protein, urate, creatinine, albumin, and trough whole blood cyclosporine concentrations. In a multiple linear regression model, only creatinine (mean 144+/-52 micromol/L; P = 0.021) and trough cyclosporine concentrations (191+/-163 microg/L; P = 0.015) were independent positive predictors of tHcy, whereas serum folate (8.35+/-7.43 nmol/L; P = 0.018) and time since transplant (P = 0.049) were significant negative predictors. We conclude that hyperhomocysteinemia is a common characteristic of cardiac transplant recipients. Our analysis suggests that folate and renal glomerular dysfunction are important contributory factors; however, whole blood cyclosporine concentrations may also predict the degree of hyperhomocysteinemia in this population and therefore influence interpretation of any apparent response to treatment.

Adult↗
PubMed1997

Pharmacodynamic monitoring of mycophenolic acid in rabbit heterotopic heart transplant model.

Pharmacodynamic (PD) monitoring of immunosuppressive drugs provides a novel approach to optimization of drug therapy in transplant recipients. We chose to investigate this using mycophenolic acid (MPA), an immunosuppressive drug that mediates its effect by the inhibition of inosine monophosphate dehydrogenase (IMPDH), a key enzyme in the de novo biosynthesis of purines. A comparison of the relationship between PD versus drug level monitoring was performed using a heterotopic cardiac transplant in New Zealand white rabbits. The animals were divided into four different treatment groups. Control animals were administered the drug vehicle, the treatment groups were administered mycophenolate mofetil (MMF) at doses of 40, 80, and 160 mg/kg/day. Statistically significant (p < 0.05) prolongation of graft survival was obtained at the 160 mg/kg/day dose group. The mean MPA concentration at this dose was approximately 2.5 mg/l, suggesting that this concentration may provide adequate immunosuppression. An increase in IMPDH activity appeared a few days prior to rejection, suggesting that measurement of enzyme activity may have potential for use as a marker of graft rejection. A significant (p < 0.05) relationship exists between MPA concentration and graft survival and the former with dose of MMF. There was a negative correlation (p = 0.17) between MPA concentration and IMPDH activity, while a trend (p = 0.37) to inverse relationship between graft survival and IMPDH activity was found. The data suggests that the measurement of the biological response may provide a useful adjunct to traditional therapeutic drug monitoring (TDM) for optimization of dosing of immunosuppressive drugs.

Animals↗
PubMed1996

Pharmacodynamic assessment of mycophenolic acid-induced immunosuppression in renal transplant recipients.

The combination of pharmacokinetic and pharmacodynamic monitoring of immunosuppressive drugs provides a novel method for the optimization of drug dosing. We chose to investigate this with the use of mycophenolic acid (MPA), an immunosuppressive drug that mediates its effect by the inhibition of inosine monophosphate dehydrogenase (IMPDH), a key enzyme in the de novo biosynthesis of purines. The relationship between MPA concentration in plasma, IMPDH activity in whole blood, and nucleotide concentration in lymphocytes was investigated in renal-transplant recipients, who were randomized to receive either mycophenolate mofetil (MMF) (n = 5) or azathioprine (AZA) (n = 7), in combination with cyclosporine and prednisone. Blood samples were collected throughout the dosing interval. Pharmacokinetic analysis revealed substantial variability among the patients in the absorption and clearance of MPA. An inverse relationship was found between the MPA concentration of IMPDH activity in whole blood. The peak concentration of MPA achieved at 1 hr after dosing resulted in approximately 40% inhibition of IMPDH activity. As the MPA concentration decreased throughout the dosing interval, there was a gradual restoration of IMPDH activity. The inhibition of IMPDH activity (P < 0.05) in MMF-treated patients as compared with the AZA-treated controls was maintained for approximately 8 hr after dosing. No statistically significant (P > 0.05) difference between the predose and the 12 hr postdose activity was observed. The concentrations of guanine nucleotides, GDP and GMP, were significantly lower than in the AZA-treated group at most of the time points after dosing; however, considerable variability was observed. The measurement of the pharmacodynamic response to immunosuppressive drugs may provide not only a mechanism to predict the most appropriate dosing regimen, but also a viable alternative to traditional therapeutic drug monitoring, by assessing the overall state of immunosuppression.

Azathioprine↗
PubMed1996

Pharmacodynamic assessment of mycophenolic acid-induced immunosuppression by measurement of inosine monophosphate dehydrogenase activity in a canine model.

The combination of pharmacokinetic and pharmacodynamic (measurement of the biological effect) monitoring of immunosuppressive drugs provides a method for the optimization of drug dosing. We chose to investigate this using mycophenolic acid (MPA), an immunosuppressive drug that mediates its effect by the inhibition of inosine monophosphate dehydrogenase (IMPDH), a key enzyme in the de novo biosynthesis of purines. Using an assay developed for measurement of IMPDH activity in whole blood, the concentration required for 50% inhibition of IMPDH activity was approximately 200 mg/L (58 +/- 8.3% for whole blood [n = 6] and 55 +/- 10.0% for isolated lymphocytes). To ascertain the relationship between MPA concentration and IMPDH inhibition in vivo, dogs were administered a single dose of mycophenolate mofetil, the pro-drug of MPA, at 20 or 40 mg/kg orally. Pharmacokinetic analysis revealed that the Cmax of the 40-mg/kg group was statistically greater than that of the 20-mg/kg group (P < 0.05). There were no statistical differences in the other parameters investigated (area under the curve, beta half-life, mean residence time, volume of distribution at steady state, and clearance) between the two treatment groups. The half-life was calculated at approximately 8 hr for both dose groups. There was also substantial variability among the dogs in the absorption and clearance of MPA. An inverse relationship was found between the MPA concentration and IMPDH. Maximal inhibition of IMPDH activity of 30-40% occurs approximately 2-4 hr after dosing, followed by a gradual restoration in enzyme activity. After 24 hr, there is an increase in IMPDH activity that exceeds the pre-dosing levels in some cases by 3-fold. Evaluation of the pharmacokinetic and the pharmacodynamic responses to MPA in the canine model suggests that the drug should be administered ever 8 hr to optimize its immunosuppressive efficacy. This combined approach can be used for optimization of doses of this and other immunosuppressive drugs.

Animals↗
PubMed1995

Pharmacodynamic assessment of mycophenolic acid-induced immunosuppression by measuring IMP dehydrogenase activity.

Pharmacodynamic monitoring of the biological effect of immunosuppressive drugs provides an alternative to traditional therapeutic drug monitoring. We chose this method to investigate mycophenolic acid (MPA), an immunosuppressive drug that mediates its effect by inhibition of IMP dehydrogenase (IMPDH), a key enzyme in the de novo biosynthesis of purines. Using an assay developed for measuring IMPDH activity in whole blood, we found the concentration of MPA required for 50% inhibition of enzyme activity to be in the range of 2.0-5.0 mg/L for both human and rabbit blood. The amount of enzyme activity in whole blood depended on the concentration of the leukocytes, was unaffected by the type of anticoagulant used, and was stable in blood specimens stored for as long as 48 h at 4 degrees C. An inverse relationship was found between plasma MPA concentrations and IMPDH activity in rabbits administered a single dose of RS-61443, the prodrug of MPA. Maximal inhibition of IMPDH activity (by approximately 60%) occurs at peak concentrations of MPA; as the concentration of the drug decreases postdose, the enzyme activity gradually increases with little or no inhibition being observed 24 h postdose.

Animals↗
PubMed1994

Comparison of the effects of cyclosporin G (OG37-325), cyclosporin A, and their metabolites on the release of endothelin and prostacyclin from primary renal and aortic endothelial cell lines.

Cyclosporin G (OG37-325) (CsG), an analogue of cyclosporin-A (CsA), has been reported in preliminary clinical trials to induce fewer renal side-effects than CsA. CsA and its metabolites have been shown to alter release of vasoactive substance both in vitro and in vivo, and this may be a contributing factor to the decreased renal blood flow and glomerular thrombosis associated with CsA-induced renal side-effects. However, the effects of CsG and its metabolites on release of such substances has not been investigated. In this study, the toxic effects of several CsG metabolites were investigated in primary cultures of rabbit endothelial and mesangial cells. The majority of the metabolites had little effect on cell growth and DNA synthesis. GM9 was the most cytotoxic metabolite, having a potency ratio of 0.18-0.36 for inhibition of cell growth. The effects of individual metabolites were examined on basal release of prostacyclin (PGI2) and endothelin (ET-1). The majority of metabolites had no effect on release of these analytes; however, GM1, at 5,000 micrograms/L resulted in a significant (p < 0.05) increase in ET-1, while GM9 at both 500 and 5,000 micrograms/L resulted in a significant (p < 0.01) decrease in PGI2 from mesangial cells. To more accurately compare the effects of CsA, CsG, and metabolites, parent drug and metabolites were added to cultures at concentrations comparable to those trough values observed at steady state in transplant recipients.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Blood distribution of mycophenolic acid.

RS-61443 (RS) a morpholinoethyl ester of mycophenolic acid (MPA), can be considered a prodrug, as immunosuppressive activity is expressed only after hydrolysis to MPA upon absorption. Little is known about the blood distribution of MPA; such information would have an impact on the medium used for analysis of the drug in clinical trials. This was investigated by spiking whole blood having an initial temperature of either 4 degrees or 22 degrees C with increasing amounts of MPA ranging from 100 to 10,000 micrograms/L. These drug concentrations span the range seen when immunosuppressive doses of the RS are administered. This was followed by incubation of the blood at 37 degrees C for 0-120 min prior to separation of the cells. The drug concentration was measured in the plasma and whole blood fractions by high-performance liquid chromatography. MPA was almost exclusively found in the plasma fraction and did not exhibit any temperature or concentration dependence. The free or unbound fraction of MPA over the same concentration range was determined by ultracentrifugation and demonstrated a concentration dependence ranging from 7.2 to 16.5% of total drug for a concentration range spanning 500-10,000 micrograms/L. The drug was found to be primarily associated with the non-albumin proteins in the plasma. Less than 10% of the drug was found to be bound to lipoproteins. The data suggest that from an analytical standpoint, plasma, rather than whole blood, would be the most suitable medium for analysis because of the higher concentrations of the drug found in this fraction.

Chromatography, High Pressure Liquid↗
PubMed1994

Steady-state concentration of cyclosporin G (OG37-325) and its metabolites in renal transplant recipients.

The steady-state concentrations of cyclosporin G (OG37-325) (CsG) and six of its metabolites (GM1, GM9, GM4N, GM1c, GM1c9, GM19) were measured throughout the 12-h dosing interval in six renal transplant recipients receiving CsG as prophylaxis against acute cellular rejection. The mean 12-h whole-blood trough concentrations (micrograms/L) were CsG, 131 +/- 26; GM1, 79 +/- 55; GM9, 110 +/- 114; GM4N, 28 +/- 18; GM1c, 31 +/- 18; GM1c9, 216 +/- 145; and GM19, 303 +/- 217. The relative concentration of the primary metabolites (GM1, GM9, GM4N) remained stable with respect to CsG throughout the dosing interval, whereas that of the secondary metabolites increased. The secondary metabolites GM19 and GM1c9 exhibited extensive between-patient variation. We investigated the effect of these metabolites on commercially available monoclonal antibody-based fluorescence polarization immunoassays (FPIA) and RIAs adapted for measurement of CsG. The 12-h whole-blood trough concentrations measured by FPIA and RIA exceed those measured by HPLC by 19% and 36%, respectively. These measured biases corresponded closely with the calculated biases (FPIA 19%, RIA 28%) based on the known cross-reactivities of CsG metabolites and their concentrations. These results suggest that cross-reactivity with metabolites account for a large part of the bias observed in immunoassays of CsG.

Antibodies, Monoclonal↗
PubMed1993

Cross-reactivities of cyclosporin G (NVa2 cyclosporin) and metabolites in cyclosporin A immunoassays.

Immunoassays of cyclosporin A (CsA) have been routinely used to measure CsG. We investigated the cross-reactivities of CsG and its metabolites, as well as the proportion CsG constitutes in relation to total drug measured, for six CsG metabolites (GM1, GM9, GM4N, GM1c, GM1c9, GM19) in the following CsA assays: Sandimmune selective RIA (SS), Sandimmune nonselective RIA (NS), Cyclotrac SP-RIA (CT), fluorescence polarization immunoassay (FPIA), and enzyme immunoassay (EMIT). The cross-reactivity of CsG in these assays was as follows: SS, FPIA, CT, approximately 100%; NS, approximately 40%; EMIT, < 2%. The cross-reactivities of CsG metabolites were investigated in all assays except EMIT and varied among metabolites and assays. The most significant variance was found with the NS assay, where most of the metabolites exhibited cross-reactivities of > 40%. In contrast, in the SS, FPIA, and CT assays, cross-reactivities of < 5% were observed for most of the metabolites. The ranking of cross-reactivities of CsG metabolites in the assays is SS = CT < FPIA < NS. The degree of cross-reactivity did not change significantly when the SS, CT, and FPIA assays were calibrated with CsG instead of CsA--whether parent CsG was present or not. The data suggest that the SS, CT, and FPIA methods would be suitable for the routine monitoring of CsG.

Cross Reactions↗