Biomedical subjects

E M Heath

Publications and source records attributed to E M Heath.

14 recordsLinked to original sources

PubMed2001

Use of buccal cells collected in mouthwash as a source of DNA for clinical testing.

CONTEXT: To maximize the participation rate in population genetic studies, alternatives to invasive whole blood collection are increasing. One such alternative is buccal epithelial cell collection, which, in contrast to venipuncture and finger sticks, is painless. Buccal cells, if collected and purified efficiently, offer an acceptable source for DNA to be used in research and clinical applications. OBJECTIVE: To develop a noninvasive sampling method for collecting cells for routine DNA testing in a clinical laboratory setting. DESIGN: Five factors were used to evaluate several brands of mouthwash: (1) compatibility with the DNA purification chemistry, (2) DNA yield, (3) DNA quality, (4) DNA stability at room temperature, and (5) mouthwash taste. Next, an optimization study was undertaken to maximize DNA yield. Finally, a validation study was undertaken with the optimized protocol to test a panel of 14 donors for DNA yield and performance and to test for the stability of DNA held in mouthwash. SETTING: Industrial research and development laboratory. RESULTS: Of 5 mouthwashes tested, Scope brand mouthwash received the highest overall ranking. The addition of proteinase K and glycogen to the protocol significantly enhanced DNA yields, with a test panel (n = 14) giving a range of 12 to 60 microg of DNA per donor. In a 4-week room temperature stability study, the DNA in mouthwash samples was found to be stable for at least 2 weeks. CONCLUSION: A clinically validated DNA purification chemistry was adapted to a noninvasive specimen collection method. This method used a commercially available mouthwash, Scope, to collect buccal epithelial cells for the preparation of high-quality DNA in high yield.

Cheek↗

PubMed1999

Effect of grip span on maximal grip force and fatigue of flexor digitorum superficialis.

The aim of this study was to investigate the effect of grip span on isometric grip force and fatigue of the flexor digitorum superficialis (FDS) muscle during sustained voluntary contractions at 60-65% of the maximal voluntary contraction (MVC). Eighteen subjects performed isometric, submaximal gripping contractions using a grip dynamometer at four different grip span settings while the pronated forearm rested on a horizontal surface. Maximal absolute grip force and median power frequency of FDS surface electromyography (EMG) during the submaximal trials were analyzed. Fatigue of FDS, as inferred from EMG frequency shifts, did not change as a function of grip size. However, middle grip sizes allowed for greater absolute forces than the small or large size. When contractions are at 60-65% MVC and the muscle is allowed to fatigue, however, grip size may be less influential than when maximal absolute force is required.

Adult↗

PubMed1999

Optimization of an automated DNA purification protocol for neonatal screening.

CONTEXT: Collection of blood from newborns is a standard clinical procedure used for genetic screening. Typically, blood from a heel prick is absorbed onto standard collection paper and dried before analysis of metabolites, proteins, hormones, and more recently DNA. OBJECTIVE: To evaluate strategies to purify DNA for use with automated workstations. DESIGN: Two factors were used to evaluate several DNA purification protocols: residual heme contamination and amplification yield. The protocol that produced DNA with the lowest heme content and the highest amplification yield was selected. In combination with those two performance factors, the protocol with the fewest number of steps was chosen to reduce reagent use and processing time. SETTING: Industrial research and development laboratory. RESULTS: Robust amplification of DNA isolated from dried blood spots was demonstrated using both fluorescence and agarose gel-based detection methods. In addition, the samples had consistent DNA volumes and had no detectable cross-contamination. Suggested instrument settings, equipment, and supplies were included for automated processing of DNA from dried blood spots. CONCLUSION: A 4-step DNA processing protocol was developed for dried blood spots. The protocol could be performed in either a manual or automated format, making it possible to process hundreds of samples in 1 day.

Blood Chemical Analysis↗

PubMed1998

Bioelectric impedance and hydrostatic weighing with and without head submersion in persons who are morbidly obese.

OBJECTIVE: To compare hydrostatic weighing with and without head submersion and bioelectric impedance analysis (BIA) for measurement of body composition of persons who are morbidly obese. DESIGN: Body composition was determined using 3 methods: hydrostatic weighing with and without head submersion and BIA. Residual volume for the hydrostatic weighing calculation was determined by body plethysmography. SUBJECTS: Subjects were 16 morbidly obese men (142.5 kg mean body weight) and 30 morbidly obese women (125.9 kg mean body weight) living in the Salt Lake County, Utah, area. Morbid obesity was defined as 40 kg or more over ideal weight. STATISTICAL ANALYSIS: One-way, repeated-measures analysis of variance was followed by Scheffé post hoc tests; body-fat measurement method served as the repeated variable and percentage of body fat as the dependent variable. Men and women were analyzed separately. In addition, degree of agreement between the 3 methods of determining body composition was determined. A regression equation was used to calculate body density for hydrostatic weighing without head submersion. Two new BIA regression equations were developed from the data of the 16 men and 30 women. RESULTS: Values for percentage body fat from hydrostatic weighing with and without head submersion (41.8% vs 41.7%, respectively) were the same for men but differed for women (52.2% vs 49.4%, respectively, P < .0001). Values for body fat percentage measured by BIA were significantly lower for men (36.1%) and women (43.1%) (for both, P < .0001) compared with values from hydrostatic weighing methods. BIA underpredicted percentage body fat by a mean of 5.7% in men and 9.1% in women compared with the traditional hydrostatic weighing method. APPLICATIONS/CONCLUSIONS: BIA tended to underpredict the measurement of percentage body fat in male and female subjects who were morbidly obese. Hydrostatic weighing without head submersion provides an accurate, acceptable, and convenient alternative method for body composition assessment of the morbidly obese population in comparison with the traditional hydrostatic weighing method. In population screening or other settings where underwater weighing is impractical, population-specific BIA regression equations should be used because general BIA equations lead to consistent underprediction of percentage body fat compared with hydrostatic weighing.

Adult↗

PubMed1998

Low energy availability, not stress of exercise, alters LH pulsatility in exercising women.

We tested two hypotheses about the disruption of luteinizing hormone (LH) pulsatility in exercising women by assaying LH in blood samples drawn at 10-min intervals over 24 h from nine young, habitually sedentary, regularly menstruating women on days 8, 9, or 10 of two menstrual cycles after 4 days of intense exercise [E = 30 kcal.kg lean body mass (LBM)-1.day-1 at 70% of aerobic capacity]. To test the hypothesis that LH pulsatility is disrupted by low energy availability, we controlled the subjects' dietary energy intakes (I) to set their energy availabilities (A = I - E) at 45 and 10 kcal.kg LBM-1.day-1 during the two trials. To test the hypothesis that LH pulsatility is disrupted by the stress of exercise, we compared the resulting LH pulsatilities to those previously reported in women with similar controlled energy availability who had not exercised. In the exercising women, low energy availability reduced LH pulse frequency by 10% (P < 0.01) during the waking hours and increased LH pulse amplitude by 36% (P = 0.05) during waking and sleeping hours, but this reduction in LH pulse frequency was blunted by 60% (P = 0.03) compared with that in the previously studied nonexercising women whose low energy availability was caused by dietary restriction. The stress of exercise neither reduced LH pulse frequency nor increased LH pulse amplitude (all P > 0.4). During exercise, the proportion of energy derived from carbohydrate oxidation was reduced from 73% while A = 45 kcal.kg LBM-1.day-1 to 49% while A = 10 kcal.kg LBM-1.day-1 (P < 0.0001). These results contradict the hypothesis that LH pulsatility is disrupted by exercise stress and suggest that LH pulsatility in women depends on energy availability.

Adolescent↗

PubMed1996

Specimen stability for DNA-based diagnostic testing.

The use of molecular diagnostic testing is increasing in the clinical setting; therefore, data regarding DNA stability in clinical specimens are essential for correct test performance and interpretation. This study was designed to determine DNA stability in peripheral blood and solid tissue under different storage conditions. DNA quality and yield were assayed by spectrophotometric absorbance, gel electrophoresis, and suitability for Southern hybridization and polymerase chain reaction (PCR), the most widely employed clinical DNA analyses. A second goal of the study was to evaluate DNA stability during storage at 4 degrees C for 1 month to 3 years. The data show that freezing or refrigeration of separated leukocytes is preferable for short- to intermediate-term storage and freezing is preferable for solid tissue. DNA degradation varying from slight to severe is seen inconsistently with such specimens, probably due to sampling of unevenly frozen-tissue areas. Depending on the degree of DNA degradation, analysis may still be possible by PCR and in some cases even by Southern hybridization. Once isolated, DNA was stable at 4 degrees C for at least 3 years. These results suggest a more flexible approach to specimen requirements for molecular pathology, as some samples that would routinely be rejected gave interpretable results.

Blood Cells↗

PubMed1995

Pregnancy weight retention in morbid obesity.

Recent hypotheses suggest that for women who develop morbid obesity, increases in weight associated with pregnancy may represent a significant contribution to their obesity status. The effects of multiple pregnancies on weight gain were studied in 96 morbidly obese women (< 13.6 kg over ideal weight at ages 20-24 or before an earlier first pregnancy and currently > 44.5 kg over ideal weight) and 115 random control women from the Utah population. Self-reported weights for each pregnancy included: prepregnancy, greatest during pregnancy, and 6 weeks following delivery, which were validated against available hospital records. Mean number of pregnancies in each group were similar (4.2 and 4.3), ranging from 1 to 9. Mean current age was 46 and mean weight gain since ages 20-24 was 46.0 kg in the morbidly obese and 14.1 kg in controls. Regression of current weight on total number of pregnancies, adjusting for weight at ages 20-24, showed a 1.3 kg/pregnancy increase in current weight (p = 0.03) with no difference between groups (p = 0.6). Weight gain subsequent to the last pregnancy was not related to the number of pregnancies (p = 0.2). Morbidly obese women gained more weight during pregnancy than controls only for the first pregnancy. Gains were similar for all other pregnancies. Morbidly obese women had smaller weight losses after delivery than the controls, but these differences were not significant. For the first pregnancy, morbidly obese women had a net weight retention that was 4.0 kg greater than the controls at 6 weeks post-partum and an average of 1.6 kg/pregnancy greater retention for the remaining pregnancies. Pregnancy weight gains for each pregnancy subsequent to the first pregnancy were constant. These findings suggest: 1) women who develop morbid obesity have slightly less weight loss after delivery and greater between-pregnancy weight gains than controls; 2) the number of pregnancies does not affect the amount of weight gained after the last pregnancy; and 3) while multiparity may augment weight gain in morbidly obese women, it is probably not a primary factor in the later development of morbid obesity.

Adult↗

PubMed1994

Induction of low-T3 syndrome in exercising women occurs at a threshold of energy availability.

To investigate the relationship between energy availability (dietary energy intake minus energy expended during exercise) and thyroid metabolism, we studied 27 untrained, regularly menstruating women who performed approximately 30 kcal.kg lean body mass (LBM)-1.day-1 of supervised ergometer exercise at 70% of aerobic capacity for 4 days in the early follicular phase. A clinical dietary product was used to set energy availability in four groups (10.8, 19.0, 25.0, 40.4 kcal.kg LBM-1.day-1). For 9 days beginning 3 days before treatments, blood was sampled once daily at 8 A.M. Initially, thyroxine (T4) and free T4 (fT4), 3,5,3'-triiodothyronine (T3) and free T3 (fT3), and reverse T3 (rT3) were in the normal range for all subjects. Repeated-measures one-way analysis of variance followed by one-sided, two-sample post hoc Fischer's least significant difference tests of changes by treatment day 4 revealed that reductions in T3 (16%, P < 0.00001) and fT3 (9%, P < 0.01) occurred abruptly between 19.0 and 25.0 kcal.kg LBM-1.day-1 and that increases in fT4 (11%, P < 0.05) and rT3 (22%, P < 0.01) occurred abruptly between 10.8 and 19.0 kcal.kg LBM-1.day-1. Changes in T4 could not be distinguished. If energy deficiency suppresses reproductive as well as thyroid function, athletic amenorrhea might be prevented or reversed by increasing energy availability through dietary reform to 25 kcal.kg LBM-1.day-1, without moderating the exercise regimen.

Adult↗

PubMed1994

Dietary restriction reduces luteinizing hormone (LH) pulse frequency during waking hours and increases LH pulse amplitude during sleep in young menstruating women.

To determine the effect of dietary energy restriction on gonadotropins, we assayed LH and FSH in samples drawn at 10- and 60-min intervals, respectively, over 24 h from seven young women (mean +/- SE gynecological age, 7.7 +/- 1.2 yr) on day 9, 10, or 11 of two menstrual cycles. Cortisol was measured in samples collected at 30-min intervals. During the 4 previous days and the day of sampling, dietary energy intake was set at either 45 or 10 Cal/kg lean body mass.day in random order. Beginning 2 days before treatment, blood was sampled daily at 0800 h and assayed for T3, insulin-like growth factor-I, and insulin. Estradiol was measured in samples collected daily and at 6-h intervals on the day of frequent sampling. By the day of frequent sampling, dietary restriction had reduced T3 20% (P < 0.01), insulin-like growth factor-I 58% (P < 0.001), and insulin 54% (P < 0.001). Twenty-four-hour transverse means for LH (P = 0.3), FSH (P = 0.2), estradiol (P = 0.3), and cortisol (P = 0.13) were unaffected, but LH pulse frequency was reduced 23% (P < 0.01), especially during waking hours, whereas LH pulse amplitude was increased 40% (P = 0.05), especially during sleep. These results support the hypothesis that LH pulsatility depends upon energy availability in women, as it does in other mammalian species.

Adult↗

PubMed1993

Effects of nicotinic acid on respiratory exchange ratio and substrate levels during exercise.

The purpose of this study was to investigate the effects of pharmacologic treatment with nicotinic acid (NA) (3 g NA.d-1) on fuel utilization during exercise and on total and HDL-cholesterol levels of trained runners. Eight experienced male runners performed four 30-min submaximal treadmill runs (SUB) at approximately 60% of aerobic capacity: the first served as the control condition, whereas SUBs 2, 3, and 4 were conducted at the onset, midpoint, and conclusion of the 3-wk NA treatment period. A 1-g NA dose was ingested 1 h prior to SUBs 2-4, which were conducted in the morning following a 12-h fast. The respiratory exchange ratio (RER) was significantly higher during SUBs 2, 3, and 4 (0.919 +/- 0.009, 0.898 +/- 0.007, and 0.896 +/- 0.009, respectively) than during SUB1 (0.871 +/- 0.008); and the RER during SUB2 was also significantly higher than that during SUB3 and SUB4 (P < 0.001). Serum free fatty acid (FFA) and glycerol levels were significantly lower during SUBs 2, 3, and 4 as compared with SUB1 (P < 0.001). Total cholesterol showed a significant decrease (from 195.3 +/- 9.2 to 174.5 +/- 9.2 mg.dl-1) and HDL cholesterol showed a significant elevation (from 56.2 +/- 2.9 to 63.0 +/- 3.9 mg.dl-1) during the NA treatment period (P < 0.05). The RER, FFA, and glycerol data demonstrate a persistent inhibition of fat utilization during submaximal exercise due to NA treatment. The 3-wk NA treatment period also proved to be sufficient to effect changes in total and HDL-cholesterol levels of these runners.

Adult↗

PubMed1991

Conservative management of primary hyperparathyroidism.

The personal management of patients seen with primary hyperparathyroidism over the past 10 years has been reviewed; 248 cases were identified of which 122 have been managed conservatively. These patients were mainly asymptomatic. Apart from 4 who developed renal stones, which were spontaneously passed, there was no clinical or biochemical evidence that patients sustained adverse consequences by conservative management. These studies support the need for further studies of the conservative management of asymptomatic hyperparathyroidism.

Adult↗

PubMed1991

Genetic and molecular analysis of repression in the P-M system of hybrid dysgenesis in Drosophila melanogaster.

Twelve inbred lines derived from an M' strain of Drosophila melanogaster were used to study the repression of P-element-mediated hybrid dysgenesis. Initial assessments indicated that the lines differed in the ability to repress gonadal dysgenesis, and that this ability was highly correlated with the ability to repress snw hypermutability. Later assessments indicated that most of the lines with low or intermediate repression potential evolved to a state of higher repression potential; however, Southern analyses failed to reveal significant changes in the array of genomic P elements that could account for this evolution. In addition, none of the lines possessed the incomplete P element known as KP, which has been proposed to explain repression in some D. melanogaster strains. One of the lines maintained intermediate repression potential throughout the period of study (52 generations), indicating that the intermediate condition was not intrinsically unstable. Genetic analyses demonstrated that in some of the lines, repression potential was influenced by factors that were inherited maternally through at least two generations; however, these factors were not as influential as those in a classic P cytotype strain. Additional tests with a dysgenesis-inducing X chromosome called T-5 indicated that repression itself was mediated by a combination of maternal effects and paternally inherited factors that were expressed after fertilization. These tests also suggested that in some circumstances, the P transposase, or its message, might be transmitted through the maternal cytoplasm.

Animals↗

PubMed1990

Genetic basis of the difference in alcohol dehydrogenase expression between Drosophila melanogaster and Drosophila simulans.

Drosophila melanogaster and its sibling species, Drosophila simulans, differ in expression of the enzyme alcohol dehydrogenase (ADH). Adult melanogaster flies that are homozygous for the Slow allozyme have approximately twice the level of ADH activity and crossreacting material as simulans adults. There is no corresponding difference in ADH mRNA, however, so this difference in ADH protein level is evidently due to a difference in the rate of translation of the two RNAs and/or to a difference in protein stability. Here we report an interspecific gene-transfer experiment, using P-element transformation, to determine whether this expression difference is due to genetic background differences between the species (trans-acting modifiers) or to cis-acting factors within the Adh gene. When the Adh genes from D. melanogaster and D. simulans are put into the same genetic background, there is no detectable difference in their level of expression. The level is relatively high in the melanogaster background and relatively low in the simulans background. Therefore, the interspecific difference in Adh expression is due entirely to trans-acting modifiers, in spite of the many sequence differences between the Adh genes of the two species, which include two amino acid substitutions.

Alcohol Dehydrogenase↗

PubMed1983

Dose determinations for waterborne 2,5,2',5'-[14C]tetrachlorobiphenyl and related pharmacokinetics in two species of trout (Salmo gairdneri and Salvelinus fontinalis): a mass-balance approach.

A mass-balance study was undertaken to evaluate the accuracy of dose determinations of waterborne 2,5,2',5'-[14C]tetrachlorobiphenyl (TCB) made on transected brook trout (Salvelinus fontinalis) and rainbow trout (Salmo gairdneri) and to determine any pharmacokinetic differences between the two species. The total calculated [14C]TCB absorbed by brook (17.8 micrograms) and rainbow (24.5 micrograms) trout was compared to the actual body burden measurements of [14C]TCB for brook (17.4 micrograms) and rainbow (25.6 micrograms) trout; the latter measurements also included excretory losses through the urine, feces, and across the gill surface. Approximately 1% of the total dose was excreted of which 75% was in the urine and 25% in the feces. The agreement between the whole body burden measurements of [14C]TCB and the total calculated micrograms of [14C]TCB absorbed was within 10% in both species. Mass-balance measurements were converted to dose by dividing by fish weight. Mean calculated and measured doses were 31.4 and 30.4 micrograms/kg/48 hr for brook trout and 32.3 and 33.6 micrograms/kg/48 hr for rainbow trout. No species differences were seen in either calculated or measured doses or in total radioactivity excreted. Respiratory function between the two species was similar except for a significantly higher mean ventilation rate for brook trout (84 +/- 14/min) than for rainbow trout (65 +/- 4/min). There were also no fluctuations noted in either respiratory function of [14C]TCB uptake efficiency across the gills over the 48-hr exposure period. These studies demonstrated the ability to accurately calculate a water dose in micrograms per kilogram per hour for individual fish that could be directly compared to other fish species or to mammals.

Animals↗