PubMed Health⌕ Search

Biomedical subjects

N N Rehak

Publications and source records attributed to N N Rehak.

At least 19 recordsLinked to original sources

Biochemical composition and electrolyte balance of "unstimulated" whole human saliva.

The biochemical composition of "unstimulated" whole saliva was determined in healthy adult subjects. Based on their relative concentration, salivary analytes could be classified into three arbitrary categories: concentration lower than in serum (saliva/serum ratio < 0.5; 12 analytes), similar to serum (ratio = 0.5-1.5; five analytes), and higher than in serum (ratio > 1.5; five analytes). Consistent with local production, an elevated lactate dehydrogenase (LDH) activity in the saliva was associated with a non-serum like LDH isoenzyme pattern: LDH5 >> LDH4 > LDH3 >> LDH2 > LDH1. Compared with serum, the concentrations of hydrogen (as reflected in the pH), potassium and inorganic phosphorus were much higher (saliva/serum ratio > or = 3), whereas that of sodium, total magnesium, chloride, and total carbon dioxide were lower (saliva/serum ratio < or = 0.3). The concentration of ionized calcium was similar in saliva and serum (saliva/serum ratio = 0.8), while ionized magnesium was unmeasurable in saliva. The salivary ionized calcium fraction was higher (0.76) than previously suggested (0.51). The difference between the main salivary cations (potassium, sodium), and anions (phosphate, chloride) was similar to serum (anion gap: 4 vs. 11 meq/l). Highly significant (p < or = 0.012) correlations occurred among salivary pH, dihydrophosphate, total calcium, and potassium. Our data suggest that calcium, potassium, chloride and phosphates are the major salivary complex-forming ions. The major compositional differences between serum and saliva show that saliva is not a passive "ultrafiltrate" of serum and salivary constituents may play a distinct physiological role.

Adult↗

Serum ionized magnesium: comparison of results obtained with three ion-selective analyzers.

In a two-center (Academic Medical Center, The Netherlands, and National Institutes of Health, USA) study, we compared ionized magnesium (iMg2+) results in serum determined with the AVL 988/4, KONE Microlyte 6 and NOVA CRT, which are the currently available analyzers equipped with a magnesium ion-selective electrode. The comparison was performed with frozen serum samples from normal individuals and patients. Imprecision and reference intervals were established. We found the best agreement between the KONE(x) and AVL(y) magnesium ion-selective electrodes (y= 0.972x-0.013; n=138) with samples from patients. With samples from normals, all three analyzers reported significantly different results (p<0.05). Best precision was found using the NOVA; coefficients of variation established at three levels were all < 4.0%. Coefficients of variation for the AVL and KONE were <5% at normal and high iMg2+, but 10.7 and 9.4%, respectively, at iMg2+ approximately 0.30 mmol/l. The reference intervals (mean+/-standard deviation) based on measurements in fresh serum samples were different for each analyzer: 0.55-0.63 mmol/l for AVL, 0.470.57 mmol/l for KONE and 0.43-0.55 mmol/l for NOVA. Thus, significant differences among the ionized magnesium concentration obtained with the three analyzers, limit comparison of results in clinical practice, and need to be resolved (e.g. by improvement of specificity and standardization of calibrators).

Adult↗

Clinically important intermethod differences for physiologically abnormal ionized magnesium results.

We compared physiologically abnormal low and high ionized magnesium (iMg) results determined with the AVL 988-4 (AVL, Graz, Austria) and Nova CRT (Nova Biomedical, Waltham, Mass) ion-selective electrodes (ISEs) in serum samples from randomly selected patients. A result of < 0.39 mmol/L with either ISE constituted the low magnesium group and of > or = 0.65 mmol/L the high magnesium group. Within each group we found significant differences between the iMg results. Major intermethod differences were found for samples with physiologically normal total magnesium concentration: most of the samples in the low magnesium group (83%) had abnormally low results with the Nova ISE, whereas most of the results with the AVL ISE (83%) were normal. In contrast, all results with the AVL ISE for the high magnesium group were abnormally high, but 67% of the results with the Nova ISE were normal. The agreement for the clinical interpretation of iMg results based on the reference interval for each method was only 32%. The differences in iMg results between the two analyzers must be resolved before using the iMg test as measured with ISE for patient care.

Blood Chemical Analysis↗

Serum ionized magnesium in chronic alcoholism: is it really decreased?

Chronic alcoholism is associated with a marked deficit in total magnesium (tMg). However, little is known about the status of the physiologically active form, ionized magnesium (iMg). We assessed serum iMg (measured with two ion-selective electrodes, AVL 988-4 and NOVA CRT) and tMg concentrations in chronic alcoholics at admission (n = 31) and after abstinence (n = 13) and compared these results with those for a control group (n = 40). At admission, the tMg and NOVA iMg concentrations in alcoholics (0.78 +/- 0.020 and 0.38 +/- 0.016 mmol/L, respectively) were significantly less (P <0.001) than in the controls (0.85 +/- 0.008 and 0.50 +/- 0.006 mmol/L). The AVL iMg results, however, did not differ significantly between the two groups: 0.53 +/- 0.013 vs 0.56 +/- 0.006 mmol/L, respectively (P >0.05). The mean iMg between the two analyzers differed significantly in both groups (P <0.001). After 3 weeks of abstinence, the alcoholics showed a significant increase in tMg (P <0.001) and in both NOVA and AVL iMg values (P <0.01 for each). tMg concentrations were positively correlated with the AVL iMg values in both alcoholics and controls but correlated positively with the NOVA iMg results only in the controls. Thus, the altered status of iMg is instrument-dependent, and the usefulness of the measurement in alcoholics is yet to be determined.

Adult↗

Thiocyanate in smokers interferes with the Nova magnesium ion-selective electrode.

Thiocyanate found in serum ordinarily is the metabolite of cyanide that is inhaled with tobacco smoke and ingested with cyanogenic foods. We investigated the effect of the thiocyanate ion (SCN-) on the ionized magnesium (iMg) and ionized calcium (iCa) results determined with the AVL and Nova magnesium and calcium ion-selective electrodes (ISEs). We analyzed saline and pooled serum with added SCN-, and serum from apparently healthy nonsmokers (n = 20) and smokers (n = 20). The mean (and range) of the measured serum SCN- concentration was 0.019 (0.008-0.046) mmol/L for nonsmokers and 0.077 (0.020-0.138) mmol/L for smokers. Only the Nova iMg results decreased with increasing SCN- concentration, and the change was dependent on the baseline iMg concentration. In the absence of Mg, SCN- decreased the voltage response of the Nova Mg ISE to calcium ions. At apparently normal serum iMg and iCa concentrations, the interference by SCN- appeared to be equimolar (iMg = -1.04 x SCN- + 0.52). Thus, the serum SCN- commonly found in smokers causes a significant (P < 0.0001) decrease in the Nova iMg results.

Artifacts↗

The effect of smoking on the serum ionized magnesium concentration is method-dependent.

OBJECTIVE: To investigate the effect of smoking on serum ionized magnesium concentration ([Mg2+]) determined by the NOVA and AVL Mg ion-selective electrodes (Mg ISEs). METHODS: Subjects were apparently healthy smokers (n = 30) and nonsmokers (n = 30). We determined NOVA and AVL [Mg2+] in their serum and in test solutions containing compounds increased by smoking. We also determined subjects' white blood cell and differential counts. RESULTS: For smokers, the mean values for NOVA and AVL [Mg2+] differed significantly (0.41 vs 0.52 mmol/L, respectively). We found a significant intramethod difference in NOVA [Mg2+] (0.11 mmol/L, P < .0001) between smokers and nonsmokers. A dose-dependent decrease in NOVA [Mg2+] was observed with an increase in cigarettes/day. NOVA [Mg2+] inversely correlated with white blood cell counts. There was no interference by the test compounds with either Mg ISE. CONCLUSION: Smoking may induce a serum factor, possibly related to white blood cells, that negatively interferes with the response of the NOVA Mg ISE.

Arachidonic Acid↗

Falsely high ionized magnesium results by an ion-selective electrode method in severe hypomagnesemia.

Changes in serum total and ionized magnesium (Mg and Mg2+) and calcium (Ca and Ca2+) were monitored in three patients who transiently developed severe (total Mg < 0.50 mmol/l) to profound hypomagnesemia (total Mg < 0.35 mmol/l) due to cisplatin or interleukin-2 therapies. Mg2+ and Ca2+ were measured with the Nova ion-selective electrodes at 37 degrees C and all results were normalized to pH 7.40. Independent of the etiology, the Mg2+ fraction (Mg2+/total Mg) increased as the concentration of the serum total Mg decreased in all three patients. When the total Mg was around or below 0.35 mmol/l the Mg2+ approached or exceeded total Mg, suggesting an error in the measurement of Mg2+. The findings were extended by including a group of 31 additional patients whose serum total Mg, Mg2+, total Ca, and Ca2+ concentrations varied from abnormally low to above normal. The serum total and ionized concentrations strongly correlated for both Mg (r2 = 0.88) and Ca (r2 = 0.92). The Mg2+ fraction rapidly increased with a fall in the total Mg concentration (r2 = 0.76) and total Mg/total Ca ratio (r2 = 0.71). In fact, with decreasing total Mg concentrations or total Mg/total Ca ratios, the Mg2+ fraction progressively increased to 93-128% of the total, confirming an error in the Mg2+ determinations. The Ca2+ fraction showed a slight and insignificant decrease with falling total Ca concentrations and total Mg/total Ca ratios. The Mg2+ concentration was directly related (r2 = 0.62), whereas the Ca2+ concentration showed a complex relationship to the total Mg/total Ca ratio. Whether this latter relationship represents a technical artifact or a true biological phenomenon requires further study. The apparent overestimation of Mg2+ at very low total Mg concentrations, and in the presence of a very low total Mg/total Ca ratio, could be due to improper chemometric correction of the Ca effect on the Mg electrode, non-linearity, and inadequate calibration. Whatever the mechanism, the failure of this method to correctly measure very low serum Mg2+ concentrations in the sera of patients with severe hypomagnesemia, or likely in any patient with an unusually low total Mg/total Ca ratio, erodes its diagnostic usefulness.

Adult↗

Comparison of precision and effect of pH and calcium on the AVL and NOVA magnesium ion-selective electrodes.

We compared the precision of the AVL 988-4 and NOVA CRT instruments for determining ionized magnesium (iMg) and assessed the effect of pH and ionized calcium (iCa) concentration on the results Within-run and day-to-day precision for the iMg electrodes were determined using three levels of control material supplied by each manufacturer. The effect of pH on iMg results was assessed by analyzing anaerobic serum samples from patients, reanalyzing those same samples after pH was increased by in vitro loss of CO2 and comparing the results. To assess the effect of iCa concentration on the iMg results, we added CaCl2 to aqueous standards from both manufacturers and to a normal serum pool. The results show comparable coefficients of variation for the two iMg electrodes both within-run (0.68-2.05 for NOVA; 0.77-2.60 for AVL) and day-to-day (2.90-6.48 for NOVA; 1.71-4.93 for AVL). The AVL results were not affected by the increase in serum pH and agreed with the NOVA results that were adjusted to a pH of 7.4 (paired t-test; p > 0.2). There was a significant direct relationship between the iCa and iMg results for both analyzers, but the AVL slopes were smaller (0.026, 0.083) than the NOVA slopes (0.129, 0.165). Thus, these two iMg electrodes have comparable precision but differ in response to an increase in pH and iCa.

Calcium↗

Linearity and stability of the AVL and Nova magnesium and calcium ion-selective electrodes.

We studied the stability and linearity of the AVL and Nova Mg and Ca ion-selective electrodes and the relation between the ionized Ca and ionized Mg results reported by each analyzer. The response of the electrodes to different concentrations of Mg and Ca was determined for saline solutions, aqueous solutions, and serum samples. The electrodes from both manufacturers demonstrated acceptable stability for the time of the study. The response of the electrodes was linear within the range specified by each manufacturer, but relative nonlinearity and the values for the linear limits differed between the AVL and Nova analyzers. The ionized Mg results varied with the concentration of Ca. The relation between ionized Ca and ionized Mg results was nonlinear and differed between the AVL and Nova electrodes. Intermethod comparison between the electrodes showed poor agreement for ionized Mg results, especially at low and high concentrations of total Ca and total Mg.

Calcium↗

Analyzer-dependent differences in results for ionized calcium, ionized magnesium, sodium, and pH.

We compared two ion-selective analyzers (AVL 988-4 and NOVA CRT) for determining ionized calcium (iCa2+), ionized magnesium (iMg2+), sodium (Na+), and pH in serum specimens from healthy and diseased individuals. For assays of three levels of protein-based control materials, total imprecision (CV) was < 3% for all analytes except iMg2+ (< or = 6.5% on NOVA, and < or = 4.9% on AVL). We found a significant difference between the analyzers (P < 0.001) for the mean iMg2+ concentration in patients but no significant correlation (r = 0.253) between the analyzers for iMg2+ in specimens from healthy volunteers, even though the mean iMg2+ concentration did not differ significantly between these groups. The reference interval (central 95 percentiles) for iMg2+ with AVL (0.44-0.60 mmol/L) was contained within that of NOVA (0.39-0.64 mmol/L). The AVL gave higher values for iCa2+ (P < 0.001) and lower values for pH (P < 0.001) in specimens from normal volunteers and patients. The mean value for Na+ in patients' samples was significantly higher by the NOVA (P < 0.01) than by the AVL analyzer. Thus, we found significant differences between these two analyzers for all four analytes.

Calcium↗

Biochemical analysis of thyroid cyst fluid obtained by fine-needle aspiration.

Despite the relatively ready availability of thyroid cyst fluid specimens, little has been published on their biochemical composition. We measured the concentrations of 18 analytes in thyroid cyst fluid specimens from benign (n = 17) and malignant (n = 3) lesions and in homogenates of normal thyroid tissue (n = 5). The concentrations of an additional five analytes were measured in selected cyst fluid specimens only. Compared with normal human serum specimens, we found that in thyroid cyst fluid specimens the activities of acid phosphatase, aspartate aminotransferase, amylase, and lactate dehydrogenase, and the concentrations of iron and total bilirubin were highly increased. The concentration of glucose was low. The gross appearance of the fluids and the presence of certain analytes were consistent with a hemorrhagic origin of most of the benign and malignant cyst fluid specimens. Other biochemical markers, however, indicated colloidlike features and/or an admixture of thyroid tissue components to the cyst fluid. Although we have limited data for cyst fluid specimens from malignant thyroid lesions, we found no evidence that the results of any of the common biochemical tests would distinguish benign from malignant lesions.

Biopsy, Needle↗

Modified magnesium method in the aca III: elimination of interference by bilirubin.

Bilirubin interferes with the Du Pont magnesium method in the aca at 510 nm. We determined that bilirubin concentrations in serum samples up to 380 mg/L did not affect the absorbance measured in the aca III at 540 nm, and therefore we modified the Du Pont setting of spectrophotometer for the magnesium method to 540 and 600 nm. Accuracy and precision of the modified method were comparable with the unmodified method and with atomic absorption spectrophotometry. For comparison, we analyzed serum samples with normal (n = 37) and increased (n = 22) bilirubin concentration with the modified method in the aca III (y) and in the atomic absorption spectrophotometer (x). The results (range 0.56-1.25 mmol/L) were in good agreement (x = y = 0.84 mmol/L, D(x - y) = -0.001, SD = 0.004, t = 0.238, P greater than 0.50) and bilirubin did not interfere with the modified method.

Bilirubin↗

Storage of whole blood: effect of temperature on the measured concentration of analytes in serum.

We measured the concentrations of 29 commonly measured analytes in fresh sera and in sera that had been stored as whole blood at seven different temperatures for 24 h. We determined the effect of storage temperature and prolonged contact with cell clot on the measured concentration of each analyte, with fresh serum as the control. Significant differences were observed for concentrations of creatinine, glucose, inorganic phosphorus, potassium, and both aminotransferases. The extent of these differences was temperature dependent. Values for the remaining 23 analytes examined were essentially unaffected by the storage.

Blood Chemical Analysis↗

Comparison of the Du Pont aca and Dow methods for determination of high-density lipoprotein cholesterol.

We compared the Du Pont aca (phosphotungstate-enzymic cholesterol) and the Dow (dextran sulfate/Mg2+-enzymic cholesterol) methods for the determination of high-density lipoprotein cholesterol (HDLC) and total cholesterol in serum from 113 patients. The aca results for both total cholesterol and HDLC were significantly greater (p less than 0.0001) than the Dow results, the aca method overestimating the HDLC concentration (mean recovery 107.2% in serum samples with values assigned by the Centers for Disease Control). The precision of the aca method for HDLC was essentially the same as that of the Dow method. Bilirubin (up to 0.17 g/L), hemoglobin (up to 4 g/L), and slight lipemia (triglycerides up to 5.4 g/L) did not interfere with the aca method.

Autoanalysis↗

Enzymic determination of free and esterified cholesterol in serum by microcalorimetry.

Concentrations of free and esterified cholesterol in serum can be determined simultaneously by measuring, with a batch-type microcalorimeter, the heat released during the coupled cholesterol esterase/cholesterol oxidase/catalase enzymic reaction. To differentiate the two forms of cholesterol, we used kinetic calorimetry: the rate of heat output due to enzymic hydrolysis of esterified cholesterol (the rate-determining reaction) was subtracted from the measured heat, the difference being the heat released during the enzymic oxidation of free cholesterol (the fast reaction). Results obtained by the kinetic calorimetric method agreed with those obtained by separate sequential end-point calorimetric determinations of free and total cholesterol. We also compared the kinetic calorimetric method with the cholesterol method of Abell and Kendall and a continuous-flow modification of the Liebermann-Burchard method (Technicon SMAC). De-biased linear-regression analysis of the data indicates acceptable agreement between the calorimetric and the Abell-Kendall methods (y = 0.98x + 11.5). The correlation between results by calorimetric and SMAC methods shows a significant proportional error (y = 1.17x - 159.4). Bilirubin (up to 200 mg/L) does not interfere with the calorimetry.

Calorimetry↗

Prospective applications of calorimetry in the clinical laboratory.

Calorimetric analysis depends on the direct proportionality between the heat changes that occur during chemical reactions and the amount of reacting substances. Potential uses of calorimetry in the clinical laboratory are discussed, with examples. The calorimetric technique does not require optically clear specimens, and if the specificity of the measured reaction is assured, calorimetry can be used for quantitative determinations of components that are present in a complex matrix system such as body fluids. Specific enzymic reactions have been used to measure substrates and enzyme activities in biological specimens calorimetrically, with precision, sensitivity, and accuracy comparable to routine photometric techniques. The application of calorimetry in the clinical laboratory is limited now by its slowness, but development of automated instruments may enable the technique to become competitive with conventional analytical techniques in the clinical laboratory.

Calorimetry↗