A high-performance liquid chromatographic method for the determination of monofluoroacetate.

A simple isocratic high-performance liquid chromatographic (HPLC) method for the quantitative analysis of monofluoroacetic acid (MFA), the toxic substance of Dichapetalum cymosum, in plant material, rumen contents (gastric contents), and liver samples is described. A suitable HPLC column that gives optimum sensitivity, accuracy, precision, and separation of MFA is identified. A C-610 organic acid analysis column at ambient temperature with 0.02M H3PO4 as an eluent and ultraviolet detection at 210 nm is utilized to quantitate MFA. Using this method, the average percentage recovery in plant material, bovine liver, and rumen samples is 94.8%, and a detection limit of 12 microg/L is achievable.

High-performance liquid chromatographic analysis with diode-array detection of bradykinin, neuropeptide K, and substance P in human plasma.

A method is described for the determination of bradykinin, neuropeptide K (NPK), and substance P in patients with atypical carcinoid syndrome. The developed method uses a combination of conventional and solid-phase extraction as well as high-performance liquid chromatographic techniques. A narrow-bore C18 column with ultraviolet detection is used (200 nm). The technique recovers bradykinin at a level of 98%, NPK at 96%, and substance P at 98% (when pure standards are dissolved) at concentration levels relevant to the atypical carcinoid syndrome. In biological samples, the recovery rate of bradykinin, NPK, and substance P drops to 88, 86, and 88% respectively. The overall analysis time is 150 min from receipt of samples. This method proves to be a valuable tool in the identification of neuropeptides and thus the diagnosis of atypical carcinoid syndrome, especially in puzzling cases with nonspecific symptoms.

Transforming growth factor alpha distribution in rectal crypts as a biomarker of decreased colon cancer risk in patients consuming cellulose.

Data from rat experimental carcinogenesis studies indicate that supplemental dietary cellulose reduces the incidence of colon cancer. Epidemiology studies also indicate that high dietary fiber reduces the risk of colorectal cancer in humans. Patients diagnosed with sporadic adenomas were entered into a randomized clinical trial to determine if supplemental dietary cellulose would reduce the patients' risk for colon cancer. Immunohistochemical staining for transforming growth factor alpha (TGF-alpha) was done on biopsies of rectal mucosa taken from patients at the time of initial polypectomy and 1 year later. Results were evaluated for utility as a surrogate end point biomarker for reduction in colon cancer risk. There was a significant decrease in the fraction of the rectal crypt cells that stained for TGF-alpha in six of seven of the patients given the cellulose supplements but in only one of six of the patients not given cellulose. Thus, whether evaluated as a group or in individual patients, there was a significant decrease in TGF-alpha in rectal crypts due to cellulose intervention, which correlated with the expected ability of supplemental dietary cellulose to decrease the risk for colon cancer. Long-term testing of the ability of dietary cellulose to reduce adenoma recurrence is under way to validate the use of TGF-alpha as a surrogate end point biomarker.

Variation in total energy expenditure in young healthy free-living men.

Interindividual and intraindividual variation in total energy expenditure (TEE) were examined in 17 healthy, free-living men (weight, 56.4 to 82.4 kg; age, 18 to 30 years). TEE over 14 days, resting metabolic rate (RMR), and body composition were measured two or three times during 77 days of fixed caloric intake using doubly labeled water, respiratory gas analysis, and isotope dilution, respectively. When individual data were averaged, TEE was most significantly related to fat-free mass ([FFM] r = .73, P = .001), body mass (r = .70, P = .002), and RMR (r = .63, P = .006). After adjusting TEE for BM, a significant inverse relation with age was found (partial r = -.52, P = .032). Stepwise regression analysis showed that 69% of individual variation in TEE was explained by BM, age, and fasting respiratory exchange ratio (RER). TEE/RMR averaged 1.73 +/- 0.25 (range, 1.38 to 2.32), and was independent of age and body composition. In 10 subjects in whom triplicate observations of TEE were performed, the average experimental variation for TEE was +/- 11.9% (range, 6.1% to 19.6%) compared with a theoretical estimate of precision of +/- 5.9% based on the reported isotope dose and analytical uncertainty. The difference between theoretical estimates of precision and observed experimental variation suggests that inherent random variation in free-living TEE is +/- 10% (ie, square root of 12(2)-6(2)) in subjects maintained on fixed caloric intake. We conclude that in young free living men (1) BM, age, and RER are important determinants of TEE; and (2) intraindividual variation in TEE is approximately +/- 10% due to fluctuations in physical activity levels within individuals over time.

Human placental transfer of zinc: normal characteristics and role of ethanol.

The fetal alcohol syndrome is primarily an impairment of growth and development. Zinc deficiency also causes abnormal fetal growth. Moreover, alcohol has been shown in some rodent studies to impair placental transport of zinc. The purpose of this investigation was to define better normal human placental zinc transport and the effects of alcohol on this process. To do this we employed the isolated perfused single cotyledon human term placental model, as well as the cultured human cytotrophoblast. In the perfused placental studies, it was shown that zinc is transferred by the placenta very slowly, about 6% of the rate of transport of antipyrine, a freely diffusible marker. The transfer is comparable in both directions, maternal to fetal and the reverse. Zinc does not cross the placenta against a zinc concentration gradient, in either direction. Rather there is good evidence of significant uptake (storage) of the zinc by the placenta on the recirculating compartment side of gradient studies. Moreover, when the perfusion fluid was low (0.2 g/100 ml) in albumin, about twice as much zinc accumulated in the perfused cotyledon and there was less zinc in the maternal compartment, as compared to perfusion with ten-fold higher (2.0 g/100 ml) albumin concentrations. Thus, ligand binding in the perfusate importantly influences placental zinc uptake. Interestingly, however, the increased placental binding of zinc did not translate into greater transfer of zinc to the fetal compartment. Thus, normal zinc transfer is slow, equal bidirectionally, and dependent on ligand binding in perfusate and placenta.(ABSTRACT TRUNCATED AT 250 WORDS)

A long-term metabolic study to assess the nutritional value of and immunological tolerance to two soy-protein concentrates in adult humans.

Seventeen healthy young adult men participated in a long-term metabolic study (11 wk) to evaluate the tolerance to and protein nutritional value of two commercially produced soy-protein concentrates. Danpro-S (nine subjects) and Danprotex-H 40 (eight subjects) (Aarhus Oliefabrik A/S, Aarhus, Denmark). Each test protein (0.8 g.kg-1.d-1) served as the sole source of dietary nitrogen. Subjects remained healthy, no problems of clinical or metabolic significance appeared, body weight remained constant, and body composition indices (lean body mass by H2(18)O dilution and creatinine excretion) and basal metabolic rate did not change. N balances fluctuated around body N equilibrium. Immunological studies confirmed the absence of any allergic responses in these subjects. It is concluded that these protein concentrates can be consumed as the sole source of dietary protein for protein nutritional maintenance and with excellent tolerance.

Clinical studies of intestinal folate conjugases.

Clinical differences between the two human intestinal mucosal folate conjugases were assessed by measurement of their activities in normal individuals and in patients with chronic diarrhea of differing causes. Intracellular folate conjugase (ICFC) was 15-fold more active than brush border folate conjugase (BBFC) in jejunal mucosa from seven obese patients undergoing elective gastric bypass surgery. The activity of ICFC was similar among normal volunteers and patients with diarrhea of unknown origin (DUO), gluten-sensitive enteropathy (GSE), inflammatory bowel disease (IBD), and the short bowel syndrome (IBD-SBS). By contrast, BBFC, sucrase, and lactase were decreased significantly in GSE, and BBFC was increased in IBD-SBS. The activity of BBFC correlated with lactase and with sucrase in the normal subjects and in patients with DUO, whereas no correlations were found with the activity of ICFC in any group. Our clinical studies confirm that ICFC and BBFC are different enzymes. ICFC is not affected by intestinal disease, whereas the activity of jejunal BBFC, like that of other brush border enzymes, is decreased by mucosal injury and is also capable of adapting to distal small intestinal disease or surgical resection.

Clinical and nutritional implications of radiation enteritis.

The clinical and nutritional significance of radiation enteritis was assessed in eight patients with chronic diarrhea which followed curative doses of radiotherapy for pelvic malignancies. Steatorrhea, found in seven malnourished patients, was ascribed to ileal disease or previous surgery, or to bacterial contamination of the small intestine. Lactose intolerance, assessed by breath hydrogen excretion after oral lactose and by jejunal lactase levels, was found in six patients. In a subgroup of five patients, the administration of two different defined formula liquid diets by nasoduodenal infusion decreased fecal fluid and energy losses by about one-half. Compared to Vivonex-HN, the infusion of Criticare-HN was associated with greater likelihood of intestinal gas production but a three-fold greater utilization of protein. Intestinal malabsorption and malnutrition in radiation enteritis has diverse etiologies. Whereas nutritional support by liquid diet limits fecal fluid and energy losses, these diets differ significantly in clinical tolerance and biologic value.

Experience with home parenteral nutrition.

Twenty-nine patients underwent courses of home parenteral nutrition therapy ranging from 1.5 to 52 months (mean 14.5 months). The primary diagnoses responsible for the requirement of home parenteral nutrition were radiation enteritis in seven patients, short bowel syndrome secondary to resection in seven, Crohn's disease in two, malabsorption states in six, and other reasons in seven. Seventeen patients (57 percent) had at least one complication. Nine patients had intravenous catheter complications and nine had metabolic complications. Nine patients have died, 11 have been successfully weaned from home parenteral nutrition, and 9 continue to receive home parenteral nutrition. Home parenteral nutrition is useful in a variety of catastrophic clinical situations, including preparation for subsequent operative therapy, temporary malabsorption, and permanent disability of the gastrointestinal tract. Complications are frequent and may be life-threatening.