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Biomedical subjects

P D Klein

Publications and source records attributed to P D Klein.

At least 73 records · Page 4Linked to original sources

Uniformly 13C-labeled algal protein used to determine amino acid essentiality in vivo.

The edible alga Spirulina platensis was uniformly labeled with 13C by growth in an atmosphere of pure 13CO2. The labeled biomass was then incorporated into the diet of a laying hen for 27 days. The isotopic enrichment of individual amino acids in egg white and yolk proteins, as well as in various tissues of the hen at the end of the feeding period, was analyzed by negative chemical ionization gas chromatography/mass spectrometry. The amino acids of successive eggs showed one of two exclusive enrichment patterns: complete preservation of the intact carbon skeleton or extensive degradation and resynthesis. The same observation was made in tissue proteins. These patterns were cleanly divided according to known nutritional amino acid essentiality/nonessentiality but revealed differences in labeling among the nonessential amino acids: most notable was that proline accretion was derived entirely from the diet. Feeding uniformly 13C-labeled algal protein and recovering and analyzing de novo-synthesized protein provides a useful method to examine amino acid metabolism and determine conditional amino acid essentially in vivo.

Journal Article↗

Effect of triple therapy (antibiotics plus bismuth) on duodenal ulcer healing. A randomized controlled trial.

OBJECTIVE: To determine whether antimicrobial therapy for Helicobacter pylori infection accelerates the healing of duodenal ulcers. DESIGN: Single-blind, randomized, controlled trial. SETTING: Veterans Affairs hospital. PARTICIPANTS: One hundred and five patients with endoscopically verified duodenal ulcers. INTERVENTION: Patients received either ranitidine, 300 mg/d, or ranitidine, 300 mg/d, plus "triple therapy" (2 g/d of tetracycline, 750 mg/d of metronidazole, and 5 or 8 bismuth subsalicylate tablets per day). Triple therapy was administered for only the first 2 weeks of ulcer treatment. MEASUREMENTS: Videoendoscopic assessment of ulcer status was done until ulcer healing was complete. Evaluations were done after 2, 4, 8, 12, and 16 weeks of therapy. MAIN RESULTS: Ulcer healing was more rapid in patients receiving ranitidine plus triple therapy than in patients receiving ranitidine alone (P less than 0.01). The cumulative percentages of patients with healed ulcers in the group receiving ranitidine plus triple therapy and in the group receiving ranitidine alone were as follows: 37% and 18% after week 2; 74% and 53% after week 4; 84% and 68% after week 8; 96% and 80% after week 12; and 98% and 84% after week 16. CONCLUSION: Combined therapy with anti-H. pylori agents and ranitidine was superior to ranitidine alone for duodenal ulcer healing. Our results indicate that H. pylori plays a role in duodenal ulcer disease.

Adult↗

Water source as risk factor for Helicobacter pylori infection in Peruvian children. Gastrointestinal Physiology Working Group.

Helicobacter pylori infection is widespread among Peruvian adults by the age of 30, but the age at which children become infected, the prevalence of disease, and the role of socioeconomic status in the epidemiology of infection are not known. We used the 13C-urea breath test to study the prevalence of infection in 407 Peruvian children from Lima, aged 2 months to 12 years, from families of low and high socioeconomic status. Peruvian children acquire H pylori early in life and the number of infected individuals increases rapidly with age; overall prevalence was 48%. H pylori infection was independent of sex, but was highly correlated with socioeconomic status; prevalence of infection was higher among children from low-income families than among those from high-income families (56% vs 32%, p = 0.001). Children whose homes had external water sources were three times more likely to be infected than were those whose homes had internal water sources. Among families with internal water sources, there was no difference in H pylori infection associated with income. Children from high-income families whose homes were supplied with municipal water were 12 times more likely to be infected than were those from high-income families whose water supply came from community wells. The findings show that the prevalence of H pylori infection is high among young Peruvian children and that the municipal water supply seems to be an important source of infection among Lima children from families of both low and high socioeconomic status.

Age Factors↗

Epidemiology of Helicobacter pylori in an asymptomatic population in the United States. Effect of age, race, and socioeconomic status.

A causative role is now accepted for Helicobacter (formerly Campylobacter) pylori in type B gastritis, and evidence is accumulating that H. pylori infection plays a major contributory role in peptic ulcer disease. Preliminary studies have reported that the prevalence of H. pylori infection increases with age, but detailed information on the prevalence of the bacteria in any defined population and on the factors that may influence the pattern of distribution remains scanty. In the present study, a sensitive enzyme-linked immunosorbent assay and a [13C] urea breath test were used to investigate the prevalence of H. pylori infection among 485 healthy asymptomatic volunteers between the ages of 15 and 80 residing in the Houston metropolitan area. H. pylori infection was present in 52%. The prevalence of H. pylori infection increased rapidly with age at 1%/yr for the overall population. The frequency of H. pylori infection was higher in blacks (70%) than whites (34%) (P less than 0.001); this difference remained after adjustments were made for age, gender, educational level, income, and use of tobacco or alcohol. H. pylori infection was independent of gender but was closely correlated with socioeconomic class. There were significant inverse correlations between age-adjusted frequency of H. pylori infection and income and between educational level and H. pylori infection. There was no association between H. pylori infection and consumption of alcohol or nonsteroidal antiinflammatory drug use or smoking. Having pets was associated with a lower frequency of H. pylori infection, but this was highly associated with higher socioeconomic status. The mode(s) of transmission of H. pylori is unknown, but the social patterns of H. pylori infection are consistent with fecal-oral transmission as one important pathway. Socioeconomic factors seem to determine the age of acquisition.

Adolescent↗

Helicobacter pylori-associated exaggerated gastrin release in duodenal ulcer patients. The effect of bombesin infusion and urea ingestion.

Recent studies have shown that the exaggerated meal-stimulated gastrin release in patients with duodenal ulcer abates after eradication of Helicobacter pylori infection. Bombesin-stimulated gastrin release was compared in 11 H. pylori-infected patients with chronic duodenal ulcer and 8 uninfected healthy volunteers both before and after therapy to eradicate H. pylori. Bombesin infusion significantly increased the gastrin release both in control subjects and in patients with duodenal ulcer. Antimicrobial therapy (bismuth, tetracycline, and metronidazole) to eradicate the H. pylori infection was associated with a significant reduction in bombesin-stimulated gastrin release in patients with duodenal ulcer (from 116.9 +/- 19 pg/mL to 69.5 +/- 7 pg/mL following 50 pmol.kg-1.h-1 bombesin; and from 158 +/- 29 to 83.4 +/- 10 following 200 pmol.kg-1.h-1 bombesin: P = 0.01 for each). Antimicrobial therapy had no effect on gastrin release in uninfected volunteers, thus excluding a nonspecific effect of antimicrobial therapy on antral G-cell function. Serum gastrin was also not increased by feeding 500 mg of urea to 5 H. pylori-infected volunteers. This suggests that access of hydrogen ion to the pH-sensitive sites governing gastrin release by mucosal ammonia produced by H. pylori urease is not a critical factor. These data suggest that exaggerated gastrin release present in patients with duodenal ulcer disease is secondary to H. pylori infection.

Adult↗

Long-term nonsteroidal antiinflammatory drug use and Helicobacter pylori infection.

This study investigates whether patients who take nonsteroidal antiinflammatory drugs are more likely to have Helicobacter pylori gastritis than age-matched individuals who do not take nonsteroidal antiinflammatory drugs, and whether patients who take nonsteroidal antiinflammatory drugs who are also infected with H. pylori are more likely to have dyspepsia, mucosal damage, or ulcers than those who are not infected. Two studies were performed, one serological and the other endoscopic, both in arthritis patients receiving nonsteroidal antiinflammatory drugs chronically. The presence of H. pylori was identified with a sensitive enzyme-linked immunosorbent assay test. One hundred eighty-three patients participated in the serological study and 75 patients in the endoscopic study. The frequency of H. pylori infection increased with age, independent of nonsteroidal antiinflammatory drug use; the age-adjusted frequency of H. pylori infection in arthritis patients paralleled that of 351 asymptomatic individuals without arthritis. The frequency of H. pylori infection increased from 30.7% in age group 21-30 years to 73.4% in age group 61-75 years. Nonsteroidal antiinflammatory drug-induced mucosal injury, either hemorrhages or erosions, was more frequent in those without H. pylori infection than with infection (61% vs. 32% for hemorrhages and 57% vs. 34% for erosions for those without and with H. pylori infection; only the difference in the frequency of hemorrhages was significant, P less than 0.05). No difference was observed in the presence of dyspeptic symptoms between those with and without H. pylori infection. These data suggest that nonsteroidal antiinflammatory drug-induced damage to the gastroduodenal mucosa does not increase the susceptibility to H. pylori infection.

Adult↗

Impact of dietary cereal on nutrient absorption and fecal nitrogen loss in formula-fed infants.

STUDY OBJECTIVE: To determine the capacity of infants to digest and absorb rice cereal and to determine the effect of cereal feeding on total energy and nitrogen absorption. SETTING: Subject's residences and the Texas Children's Hospital Clinical Research Center, Houston. PATIENTS: Eight healthy 1-month-old bottle-fed infants. INTERVENTIONS: Infants were fed their usual formula for 3 days. For the subsequent 6 days, they received 4 gm of rice cereal, labeled with carbon 13, per 30 ml of the formula. MEASUREMENTS AND MAIN RESULTS: Fecal balance studies were performed for a 72-hour period while the infants received only formula and again during the last 3 days of cereal feeding. Breath samples for hydrogen measurement were collected before and after the cereal feeding. Nutrient intake was measured and stools were analyzed for 13C abundance, energy, nitrogen, fiber content, and bacterial mass. Cereal absorption was 88 +/- 9% (mean +/- SD). Despite a significant increase in energy and nitrogen intake from cereal feeding, the coefficient of absorption fell (energy: 97% to 90%, p = 0.048; nitrogen: 94% to 74%, p = 0.009). Fecal dry weight increased after the cereal feeding (p = 0.004), primarily as a result of a sevenfold increase in fecal bacterial mass (p = 0.002). Fecal nitrogen increased primarily because of incorporation of nitrogen into bacteria. No differences were detected in breath hydrogen as a consequence of formula feeding versus formula-cereal feeding. CONCLUSIONS: Although cereal was relatively well-absorbed in this group of infants and increased their intakes of energy and nitrogen, it did not increase the coefficients of energy and nitrogen absorption. Cereal feeding increased fecal bacterial mass and bacterial nitrogen.

Bacteria↗

Absorption of carbon 13-labeled rice in milk by infants during acute gastroenteritis.

To determine whether rice cereal could be used to complement a cow milk-based diet in the nutritional management of infants with acute diarrhea, we assessed its digestion and absorption in eight affected male infants, 69 to 131 days of age. They received cow milk formula with 5.4% lactose (diluted 1:1 with water and precooked rice cereal) 5 to 22 hours after admission and rehydration. The first feeding consisted of milk diluted with carbon 13-enriched rice cereal. A 48-hour fecal collection and balance study was performed. Rice cereal was reasonably well absorbed (84.0% to 95.8%) by seven of the eight infants. The study was repeated in seven of the infants after they had recovered. Our results indicated that rice cereal is well absorbed by young infants with acute diarrhea and that it is an adequate nutrient supplement for this patient population.

Acute Disease↗

Measurement of milk intake: tracer-to-infant deuterium dilution method.

The tracer-to-infant deuterium dilution method for the measurement of milk intake was evaluated in twenty breast-fed and twenty formula-fed infants. The isotope method was compared with conventional direct-weighing techniques. Human milk intake was assessed by 5 d test-weighing. Intakes of formula, supplemental foods, and water were determined by pre- and post-weighing of feeding bottles. An oral dose of 200 mg 2H2O/kg body-weight was given to each infant, and urine was sampled daily for 14 d. 2H enrichment of the urine was measured by gas-isotope-ratio mass spectrometry. Milk intakes estimated from the deuterium dilution method were consistently higher than those from direct-weighing; the mean difference between methods was 106 (SD 47) g/d or 14% for the breast-fed group and 70 (SD 155) g/d or 8% for the formula-fed group. Estimates of intake for some infants varied substantially between the two methods of measurement. When the estimated values of human milk intake were corrected for environmental water influx and insensible water loss during breast-feeding, the relative bias decreased to 5%. Correction of the estimated values of formula intake for environmental water influx decreased the relative bias to 1-2%. The acceptability of the deuterium dilution method to determine milk intake depends on the goals and the tolerance for error in group and individual intake estimates of a given study.

Body Height↗

Energy requirements of breast-fed infants.

Current recommendations for energy intake are reviewed in light of emerging data on energy intakes of breast-fed infants and on total daily energy expenditure of infants. For determination of energy requirements, the historical approach based on observed intakes of healthy infants is compared with a newly proposed approach based on energy expenditure and deposition. A data set of exclusively breast-fed infants is used to illustrate the circuity of either approach. Energy intake, expenditure, and stores must be evaluated in an assessment of dietary adequacy. Ultimately, energy requirements of infants should reflect growth rate, body composition, and level of physical activity conducive to optimal health and neurobehavioral development.

Basal Metabolism↗

Rapid preparation of pyrogen-free 2H2(18)O for human-nutrition studies.

We describe a compact ultrafiltration system for the removal of pyrogens and bacteria from water labeled with the stable isotopes of deuterium and oxygen-18. The ultrafiltration system is constructed from readily available commercial components and can achieve complete removal of pyrogens and bacteria from 1L contaminated water within 30 min. By use of our procedure, loss of the isotopically labeled water by retention in the filtration system was minimal. The purified water is suitable for both oral and intravenous administration to healthy human subjects participating in nutrition studies.

Deuterium↗

The importance of alpha-lactalbumin in infant nutrition.

The ideal "humanization" of milk substitutes should include the creation of an amino acid pattern closely resembling that of human milk. Because the mixture of proteins in human milk is particularly rich in tryptophan and cysteine and low in methionine, this pattern is difficult to achieve with commercially available proteins. Even whey-predominant formulas only approximate human milk. Human milk has a high concentration of whey protein (70% of total protein). Of this, alpha-lactalbumin, a component of the lactase synthetase complex, accounts for 41% of the whey and 28% of the total protein. Only 3% of the protein in bovine milk is alpha-lactalbumin. Human and bovine alpha-lactalbumin share a 72% amino acid sequence homology. Both proteins contain (wt/wt) 6% tryptophan and 5% cysteine but only 0.9% methionine. Thus the differences in the amino acid compositions of bovine and human milks are largely attributable to differences in their alpha-lactalbumin contents. Commercial availability of bovine alpha-lactalbumin would allow the construction of infant formulas with amino acid compositions that are very close to that of human milk. alpha-Lactalbumin would also be a valuable constituent of diets for patients whose protein intake must be restricted.

Amino Acids↗

Transmission of Helicobacter pylori infection. Studies in families of healthy individuals.

Helicobacter pylori is accepted as the commonest cause of type-B gastritis. Detailed information about the mode of transmission remains scanty. We investigated the frequency of H. pylori infection within families, defined as consisting of a husband and wife with at least one biologic child, all living in the same household. Inclusion criteria required that both the parents and the children had been born in the United States, had used no antibiotic or bismuth for the previous 2 months, had no recent major illness or surgical operation, and had no symptoms referable to the upper gastrointestinal tract. H. pylori infection was identified with a 13C-urea breath test and an enzyme-linked immunosorbent assay for anti-H. pylori IgG. Forty-one families (151 healthy individuals) were enrolled. Before the results of the H. pylori tests were known, one parent was selected as the index subject. H. pylori infection clustered; that is, 68% of spouses of H. pylori-infected index subjects were also H. pylori-infected, compared with 9% of spouses of H. pylori-negative index subjects (p less than 0.0001). The children of infected index parents were also more likely to be infected than children of uninfected index parents--40% versus 3%, respectively (p less than 0.0001)--and the results in the children were independent of whether the father or the mother was the index subject. Clustering of H. pylori infection within families suggests person-to-person transmission or common source exposure. The high frequency of H. pylori infection in spouses suggests that genetic factors are less important than living conditions for transmission of H. pylori infection.

Adolescent↗

Simple noninvasive method to test efficacy of drugs in the eradication of Helicobacter pylori infection: the example of combined bismuth subsalicylate and nitrofurantoin.

Eradication of Helicobacter pylori infections has proved to be difficult. There is a need both for improved therapies and for ways to rapidly identify therapies that show sufficient promise to be worth pursuing. The objectives of this study were to evaluate the value of a therapeutic regimen of a bismuth salt plus nitrofurantoin for eradication of infection by H. pylori and to determine the validity/utility of the urea breath test in monitoring the progress of a clinical trial. We used an 80% eradication rule to define a promising therapeutic regimen, i.e., a regimen that eradicated the infection (no evidence of infection by H. pylori 4 wk after termination of therapy) in at least 80% of the individuals treated. Eighteen men (median age 38) with documented infection by H. pylori completed the study. At the end-of-study evaluation, H. pylori infection was eradicated (negative urea breath test, culture, and histology) in only one of 18 (5.5%) subjects; 15 were positive by the urea breath test, 16 by culture, 15 by Warthin-Starry stain, and 16 by the presence of acute-on-chronic inflammation. Using the 80% eradication rule, any one of these tests alone would have identified that the combination of antimicrobials tested was not effective in the eradication of the infection. We conclude that the urea breath test is a simple, noninvasive, cost-effective method to separate promising from unpromising candidate therapies and for the evaluation of new therapeutic concepts.

Adult↗

Lysine and protein metabolism in the young lactating woman.

Five lactating and five postpartum non-lactating women of similar ages, times postpartum, body weight and height consumed a liquid formula diet that supplied 1.3 g protein and 32 kcal/kg per day (lactating subjects) and 1.1 g protein and 26 kcal/kg per day (non-lactating subjects). Their last meal supplied 25% of the daily intake and was consumed 4 h before they received L-[13C1]lysine (27 mumol/kg) by a single intravenous injection and L-[15N2]lysine (27 mumol/kg) orally. Frequent plasma and breath samples were collected for 6 h during which time they consumed no food. On a separate day, subjects received NaH13CO3 (10 mumol/kg) as a single intravenous dose and breath samples were collected for 6 h. Plasma tracer lysine levels were determined by gas chromatography-mass spectrometry isotope ratiometry, and breath 13CO2 levels were measured by gas-isotope-ratio mass spectrometry. Averaged tracer data for the two groups were fitted to a multicompartmental model of lysine and protein metabolism that partitioned lysine kinetics between a central and two tissue compartments. The tissue compartments had characteristically fast and slow rates of lysine turnover. The results were compared with those previously obtained in nulliparous women. The postpartum state was associated with a reduction in protein turnover in a compartment with a rapid rate of protein turnover and postpartum women catabolized significantly less lysine than nulliparous controls. Lactating women catabolized slightly more lysine than the non-lactating postpartum subjects, especially when lysine catabolism was expressed as a proportion of lysine flux. Lactation was associated with smaller splanchnic and extracellular pools of free lysine and with an increase in the rate constant for absorption of orally administered lysine. Lysine flux was significantly lower in the lactating subjects and this was associated with a decrease in the rate of lysine turnover in the slowly turning over lysine compartment. The results suggest that lactation is associated with a slower rate of protein turnover in a peripheral tissue compartment. We conclude that an intake of 1.3 g protein/kg per day may be inadequate to support the protein needs of lactation and body protein metabolism and may result in metabolic adaptations that maintain lactation at the potential expense of other aspects of maternal protein turnover.

Adult↗

Measurement of in vivo cholesterol synthesis from 2H2O: a rapid procedure for the isolation, combustion, and isotopic assay of erythrocyte cholesterol.

A rapid preparative scale purification of erythrocyte free cholesterol has been developed for measurements of in vivo cholesterol synthesis from 2H2O. The quantity and purity of cholesterol obtained is suitable for combustion, zinc reduction of the water formed, and determination of deuterium isotopic content by gas isotope ratio mass spectrometry. The ability to detect and to quantitate a range of cholesterol synthesis rates is illustrated by measurements on young pigs receiving diets without and with added dietary cholesterol.

Animals↗

Incorporation of a stable isotopically labeled amino acid into multiple human apolipoproteins.

Procedures are presented for the separation and determination of the isotopic enrichment of multiple human apolipoproteins labeled in vivo with a stable isotope amino acid. The isotopic enrichments of plasma lysine and plasma apolipoproteins were monitored for 16 days after a single intravenous dose of [4,4,5,5-2H4]lysine (5 mg/kg body weight). The use of a multiply deuterated amino acid enabled the measurement of isotopic enrichments above background over the entire 16-day time course in all proteins. Individual apolipoproteins were separated on a specially designed gradient sodium dodecyl sulfate polyacrylamide gel electrophoresis system cast in a conventional slab gel apparatus which resolved apoB-100, apoE, apoA-I, apoA-II, apoC-I, apoC-II, apoC-III-1, and apoC-III-2 on a single gel. After staining with Coomassie blue, proteins bands (containing 5 to 30 micrograms of individual apolipoprotein) were excised from the gel. Amino acids were recovered from hydrolyzed gel slices, derivatized, and analyzed by gas chromatography-mass spectrometry for determination of lysine isotopic enrichments. The utility of the method is demonstrated using examples of apolipoproteins B-100, A-I, A-II, C-I, C-II, and C-III from either total plasma d less than 1.21 g/ml lipoproteins or selected lipoprotein subfractions. Lysine isotopic enrichments of proteins were generally determined with a precision of better than 5%. The isotopic enrichment profiles were consistent with literature reports of apolipoprotein metabolic kinetics based on the use of radioiodinated apolipoproteins. The procedures outlined can be used to separate and measure the isotopic enrichment of virtually any apolipoprotein from any chosen lipoprotein fraction. Thus, these procedures should find wide application in the study of apolipoprotein metabolic kinetics.

Adult↗