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Biomedical subjects

J W Finley

Publications and source records attributed to J W Finley.

At least 37 records · Page 2Linked to original sources
PubMed1999

The chemical form of selenium influences 3,2'-dimethyl-4-aminobiphenyl-DNA adduct formation in rat colon.

There is increasing evidence that selenium can protect against tumorigenesis or preneoplastic lesion development induced by chemical carcinogens. This study examined whether selenite, selenate or selenomethionine would be protective against 3, 2'-dimethyl-4-aminobiphenyl (DMABP)-DNA adduct formation in the liver and colon of rats and sought to delineate the mechanism for the protective effects of the different chemical forms of selenium against aberrant crypt formation, a preneoplastic lesion for colon cancer. After injection of DMABP, two DNA adducts were identified in the liver and colon of rats. Supplementation with either 0.1 or 2.0 mg selenium/kg diet as either selenite or selenate but not selenomethionine resulted in significantly fewer (53-70%; P < 0.05) N-(deoxyguanosin-8-yl)-(deoxyguanosin-8-yl)-3, 2'-dimethyl-4-aminobiphenyl (C8-DMABP)-DNA adducts in the colon but not the liver than in rats fed a selenium-deficient diet. Rats supplemented with selenomethionine had greater (P < 0.05) plasma and liver selenium concentrations and glutathione peroxidase activity than those supplemented with selenite or selenate; however, they also had more DMABP-DNA adducts. The protective effect of selenite and selenate against DMABP-DNA adduct formation apparently is not a result of alterations in plasma or liver selenium concentrations or altered glutathione peroxidase or glutathione transferase activities but may be related to differences in the metabolism of the different forms of selenium.

Aminobiphenyl Compounds↗
PubMed1999

The retention and distribution by healthy young men of stable isotopes of selenium consumed as selenite, selenate or hydroponically-grown broccoli are dependent on the isotopic form.

Twenty-seven healthy young men were randomly assigned to diets that supplied low (32.6 microg/d) or high (226.5 microg/d) levels of selenium for a 105-d study. After consuming the diets for 85 d, subjects were fed a test meal that contained 74Se in the form of selenite or selenate and 82Se incorporated into hydroponically-raised broccoli. Urine, fecal and blood samples were collected daily. Isotope absorption was not different (P > 0.05) for selenate and Se in broccoli; Se absorption from selenite was highly variable and was not included in statistical analyses. Significantly more isotope was absorbed by subjects fed the high Se diet (P = 0. 015). Urinary isotope excretion was greater when selenate was fed than when broccoli was fed (P = 0.0001), and consequently more Se from broccoli (as compared to selenate) was retained (59.2 +/- 2.4 and 36.4 +/- 4.6% for Se in broccoli and selenate, respectively; P = 0.0001). Despite the higher retention, less isotope from broccoli than from selenate was present in the plasma. Plasma proteins separated by gel permeation chromatography showed that most of the isotopes were distributed between two medium molecular weight peaks. Less isotope was found in plasma proteins of subjects fed the high Se diet, but the form of Se had no effect on isotope distribution. These results show that dietary Se intake alters the retention of stable isotopes of Se and that humans retain and distribute Se from broccoli in a different manner than Se from inorganic salts.

Adult↗
PubMed1998

Effect of biliary ligation on manganese accumulation in rat brain.

Neurologic and radiologic disorders have been reported to occur in miners inhaling manganese (Mn)-laden dust and in humans receiving long-term parenteral nutrition. These abnormalities have been attributed to Mn intoxication because of elevated serum Mn concentrations. Because the liver, by way of the bile, is the major route of Mn excretion, it is possible that anything that decreases biliary excretion could increase accumulation of Mn in the brain. The purpose of this study was to determine whether biliary ligation would increase Mn accumulation in the brain of rats that were exposed to deficient or adequate amounts of dietary manganese. The first experiment had a 2 x 3 factorial design, two levels of Mn (0 or 45 microg/g diet) and three surgical treatments (control, sham, or bile-ligation). Animals were sacrificed 10 d after being fed 54Mn. In experiment 2, animals that had a sham operation or bile-ligation were sacrificed at 8 time points after being injected intraportally with 54Mn complexed to albumin. The biliary-ligated animals had a significantly (p < 0.001) smaller percentage of the 54Mn in their brains (when expressed as a percentage of whole animal 54Mn) than the sham-operated animals. Mn deficiency had a similar effect. However, we did observe an increased accumulation of the radioisotope in the brain over time. Therefore, in short-term studies, biliary-ligated rats do not appear to be a good model for Mn accumulation in the brains of people with cholestatic liver disease.

Animals↗
PubMed1998

Effect of selenium supplements on the distribution of selenium among serum proteins in cattle.

The objective of this study was to determine the effects of the amount and chemical form of dietary Se on the distribution of Se among serum proteins. Six growing calves were assigned in a completely randomized design to receive diets containing either adequate (0.41 microgram/g) or excess (0.73 microgram/g) dietary Se. Proteins in serum collected from the calves were separated into albumin, glutathione peroxidase, and selenoprotein P fractions, and the concentration of Se in each was determined. The concentration of Se within serum was elevated by dietary Se supplementation. The selenoprotein P fraction within serum contained the largest percentage of Se among the serum proteins. In a second study, 12 mature cows were assigned to receive one of four experimental salt mixes containing 20, 60, or 120 micrograms of Se as sodium selenite/g of salt mix; the fourth treatment was 60 micrograms of Se as selenized yeast/g of salt mix. Cows given salt with 120 micrograms of Se as selenite or 60 micrograms of Se as selenized yeast had the highest concentrations of Se in whole blood; however, concentrations of Se in serum did not differ among treatments. Concentrations of Se in the protein fractions within serum were not affected by treatment. Within serum, the highest concentration of Se was in the selenoprotein P fraction (31.6 ng/ml), the smallest concentration was in the glutathione peroxidase fraction (4.7 ng/ml), and an intermediate amount of Se was obtained from the albumin fraction (8.5 ng/ml). In conclusion, selenized yeast and selenite as sources of Se for supplementation of cattle resulted in similar patterns of Se distribution among proteins in serum. The greatest concentration of Se was found in the selenoprotein P fraction, which may contribute to Se transportation or function as an antioxidant.

Animals↗
PubMed1998

Manganese uptake and release by cultured human hepato-carcinoma (Hep-G2) cells.

The liver is the primary organ involved in manganese (Mn) homeostasis. The human hepato-carcinoma cell line, Hep-G2, shows many liver specific functions. Consequently, Hep-G2 cells were investigated as a possible model of hepatic metabolism of Mn. Initial experiments showed that the concentration of Mn in the diet, or culture medium, similarly affected the retention of Mn by isolated rat hepatocytes and Hep-G2 cells. Manganese uptake by Hep-G2 cells suggested that uptake was followed by release from the cell. Uptake was saturable and half-maximal at 2.0 micromol Mn/L, and was inhibited by iodoacetate, vanadate, cold, and bepridil. The cations Fe2+, Cu2+, Ni2+, Cd2+, and Zn2+ decreased Mn uptake. Uptake was dependent on Calcium (Ca) concentration in a manner that resembled saturation kinetics. Cells that were pulsed with 54Mn and then placed into nonradioactive medium quickly released a large portion of their internalized Mn. Release of internalized Mn could be inhibited by low temperature, nocodozole, quinacrine and sodium azide. These data show that Hep-G2 cells are a potentially good model of hepatic Mn metabolism. Mn is taken up by a facilitated process that may be related to Ca uptake. Release apparently is an active, controlled process, that may involve microtubules and lysosomes.

Animals↗
PubMed1997

A surgical model for determination of true absorption and biliary excretion of manganese in conscious swine fed commercial diets.

Some trace elements, such as Mn, Cu and Zn, are absorbed and quickly resecreted into the gut through the bile. When this occurs, the unabsorbed nutrient and the absorbed and resecreted nutrient may mix in the gut, preventing quantitative calculation of either. We have developed a surgical model that prevents this complication. Pigs (20-40 kg) were fitted with cannulas in the bile duct, lumen of the duodenum, portal vein, ileocolic vein and jugular vein. After recovery for 6-8 d, pigs were given an oral dose of 9.25 mBq of 54Mn. The flow rate of blood past the portal vein was determined by infusion of P-amino hippuric acid into the ileocolic vein. Absorption was quantified by multiplying the concentration of 54Mn in the portal blood by the flow rate. Biliary excretion was determined by quantitative collection of bile, and previously collected bile was reinfused into the gut lumen. Urine and feces were also quantitatively collected. A postoperative time of 6-8 d was sufficient for pigs to recover from the effects of surgery and anesthesia, as assessed by several measures of metabolic function and food and water intake. True absorption was calculated to be 0.5%. 54Mn in the urine and bile began to increase after 4 d. When the pigs were killed after 12 d, only 0.5% of the 54Mn remained in the carcass. Results of this study show that pigs surgically modified by the described procedure can recover fully and can serve as a model to study intestinal absorption and biliary excretion of nutrients. Furthermore, initial studies using 54Mn showed that the model is applicable to studying Mn metabolism and suggest the need for a more detailed study of Mn absorption and biliary excretion.

Animals↗
PubMed1997

The influence of culture time and passage number on the morphological and physiological development of Caco-2 cells.

The Caco-2 cell line is used by many investigators as a model of the intestinal epithelium to study nutrient uptake and transport. Our goal was to create an awareness of inherent variabilities in the Caco-2 cell line which may influence their suitability as a model or their application to specific problems. To study the influence of passage on the model, cultures were monitored from passage 20 to 109. Transepithelial electrical resistance (TEER) and sucrase activity (measured in 21-day-old cultures) increased through about passage 36. TEER values declined after about passage 60; sucrase remained elevated but variable. Cells at passage 22, 33, and 72 were grown simultaneously for 24 days. Older-passaged cells reached plateau phase sooner. Before Day 15, passage 72 cells had higher TEER and lower permeability to 14C-mannitol than passages 22 and 33; however, after Day 15 all passages showed similar permeability. On Day 21, passage 72 cells had significantly lower alkaline phosphatase activity than did the other passages. Electron microscopy did not reveal any major morphological differences between the passages; however, it did show that some areas of cells grown on membranes were not monolayers but were several cells thick with varied morphology. Investigation of the formation of these multilayered areas showed them to be an inherent part of cell growth under the conditions used. These results emphasize the inherent variability in Caco-2 cell models and emphasize the need to monitor closely the culture characteristics during growth and differentiation under specific experimental conditions.

Alkaline Phosphatase↗
PubMed1995

Use of stable isotopic selenium as a tracer to follow incorporation of selenium into selenoproteins.

Stable isotopes of selenium (Se) have been used in human studies to measure Se absorption, retention and excretion. The purpose of this study was to examine whether stable Se could also be used to follow the incorporation of Se into selenoproteins and whether selenoproteins are labeled with stable isotopes the same way they are with radioactive Se. Rats fed either a Se-deficient or a high-Se diet were injected with either a radioactive (75Se) or a stable isotope of Se (77Se), and the liver cytosol was chromatographed on Sephadex G-200. Compared with 75Se, a greater percentage of 77Se was incorporated into cytosol, but the distribution and the effect of dietary Se was similar for both isotopes. New Zealand long-eared rabbits were also injected with either 77Se or 75Se, and the plasma was chromatographed. More of the 75Se was incorporated into the plasma, but again the patterns of incorporation were similar for both isotopes. Plasma from a male subject who ingested 60 micrograms of 77Se was chromatographed, and the stable Se was detected in column fractions and showed a distribution similar to that observed for rabbit plasma. Finally, a polyacrylamide gel electrophoresis (PAGE) method was developed that allowed loading of sufficient protein to analyze for 77Se in individual protein fractions. The distribution of 77Se and 75Se in rabbit plasma was similar. Human plasma was electrophoresed by a similar method and peaks of 56 and 23 kDa were detected. These data show that stable isotopes of Se can be used for selenoprotein production in the same way as radioactive isotopes. They also show that, when physiological amounts of stable Se are ingested by humans, the isotope can be detected in blood-borne proteins separated by column chromatography and PAGE.

Adult↗
PubMed1994

Effect of bile/pancreatic secretions on absorption of radioactive or stable zinc. In vivo and in vitro studies.

Biliary/pancreatic (B/P) secretions are a major component of endogenous secretions, and endogenously secreted Zn is a primary means of Zn homeostasis. This study examined whether B/P fluid alters the absorption/reabsorption of Zn and, in doing so, whether this contributes to homeostatic control of Zn. Animal experiments utilized rats fed 10 or 300 micrograms Zn/kg diet. An open-ended gut perfusion study in which 65Zn-labeled B/P fluid or 67Zn-labeled and digested diet found significantly decreased Zn absorption from B/P fluid. Although Zn absorption from both sources was less in animals fed diets higher in Zn, there was no interaction of treatment and diet. Further studies utilizing cultured human colon carcinoma cells (CACO-2) as in vitro models of gut enterocytes found that the presence of B/P fluid significantly decreased Zn retention and/or transport and resulted in a redistribution of cellular Zn after 1200 min of incubation. These studies show that a substance in B/P fluid can decrease the absorption of Zn and also suggest that dietary Zn and Zn associated with B/P secretions are absorbed from distinct pools. However, the lack of an interactive effect with diet, and the amount of time required to see differences in CACO-2 cells, suggest that differences in absorption are not a major contributor to Zn homeostasis.

Absorption↗
PubMed1994

Characterization of biosynthetic IgG oligomers resulting from light chain variable domain duplication.

We have characterized a human IgG1 monoclonal antibody composed of altered light chains. Each light chain consists of two identical variable domains and a kappa constant domain, in association with a normal gamma chain. This antibody assembled biosynthetically into a mixture of stable oligomers and monomers. Employing gel filtration, PAGE, and electron microscopy, we examined the antibody and the nature of the associations involved in oligomer formation. By engineering a protease factor Xa site between the duplicated light chain variable domains and examining the fragments produced following factor Xa cleavage, we demonstrated the association of the IgG monomers occurred through their duplicated VL domains. Electron microscopy showed the oligomeric antibody to be predominantly dimers and trimers in which the monomeric units were associated through the tips of the Fab portion of the antibody, presumably through the protruding N-terminal VL domains. Similar examination of monomers demonstrated several molecular forms, including individual molecules with self-crosslinked Fab arms and others displaying the open Y and T shapes typically observed for IgG antibodies. The monomers also displayed distally protruding domain-like structures. The oligomers produced by this cell line therefore occurred through the noncovalent interaction between the extra light chain variable domains.

Amino Acid Sequence↗
PubMed1994

Sex affects manganese absorption and retention by humans from a diet adequate in manganese.

Men (n = 20) and women (n = 20) consuming a diet adequate in manganese were fed 0.037 mBq 54Mn in a test meal. Subjects were counted in a whole-body counter for 70 d to determine whole-body retention of 54Mn. Data from days 10 to 20 and from days 19 to 70 were analyzed by linear regression to calculate absorption and biological half-life. Men absorbed significantly less 54Mn than women, but the 54Mn absorbed had a longer half-life in men. Estimates of absorption were higher, and estimates of half-life were lower, when data from days 10 to 20 were used compared with days 19 to 70. There was a significant association between manganese absorption and plasma ferritin concentrations and between manganese absorption and biological half-life. We conclude that men and women differ in manganese metabolism and that such differences may be related to iron status. We also conclude that regression estimates of absorption determined by using whole-body retention curves depend on the portion of the data used.

Adolescent↗
PubMed1991

Effect of light chain V region duplication on IgG oligomerization and in vivo efficacy.

A human immunoglobulin G1 (IgG1) antibody oligomer was isolated from a transfected myeloma cell line that produced a monoclonal antibody to group B streptococci. Compared to the IgG1 monomer, the oligomer was significantly more effective at protecting neonatal rats from infection in vivo. The oligomer was also shown to cross the placenta and to be stable in neonatal rats. Immunochemical analysis and complementary DNA sequencing showed that the transfected cell line produced two distinct kappa light chains: a normal light chain (Ln) with a molecular mass of 25 kilodaltons and a 37-kilodalton species (L37), the domain composition of which was variable-variable-constant (V-V-C). Cotransfection of vectors encoding the heavy chain and L37 resulted in production of oligomeric IgG.

Animals↗
PubMed1991

Effect of sex and time of sampling on selenium and glutathione peroxidase activity in tissues of mature rats.

Sprague-Dawley rats were used to investigate variations in measures of glutathione peroxidase (GSH-Px) and selenium (Se) concentration resulting from diurnal cycles and sex. Mature rats (equal numbers of males and females) were killed at 4 h intervals over a 48 h period (0200, 0600, 1000, 1800 and 2200 h each day). Selenium and GSH-Px were measured in plasma, erythrocytes, and liver and kidney cytosols. Selenium concentrations did not vary diurnally, but plasma GSH-Px activities were higher during the light than dark periods. Males had greater plasma GSH-Px activities and Se concentrations (42 EU and .45 mg/kg, respectively) than females (35 EU and .41 mg/kg respectively). GSH-Px activities were also higher in male kidney cytosols than females (117 and 76 EU, respectively). Selenium and GSH-Px activities, however, were lower in male liver cytosols (.48 mg/kg and 272 EU) than females (1.19 mg/kg and 795 EU, respectively). These data suggest that Se is distributed differently in male and female rats and the difference in Se distribution is accomplished by differences in GSH-Px activities.

Animals↗
PubMed1989

Improved wheat for baking.

To bakers, wheat quality means the performance characteristics of the flour milled from the wheat when used in specific wheat products. The tremendous increase in the number of wheat cultivars grown in the U.S. in recent years, along with the unusual climate, new advances in milling technology, and increased automation of baking lines, have resulted in bakery production problems partly attributed to wheat flour quality. In this review various factors affecting wheat quality are explained. Concerns of bread and cookie/cracker manufacturers on deterioration of the wheat quality are discussed, and, finally, some solutions are proposed.

Bread↗