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PubMed2003

Effect of linseed oil supplementation on ruminal digestion in dairy cows fed diets with different forage:concentrate ratios.

The effect of linseed oil (LSO) supplementation on total-tract and ruminal nutrient digestibility, N metabolism, and ruminal fluid characteristics was investigated in dairy cows fed diets containing different forage to concentrate ratios (F:C). The experimental design was a 4 x 4 Latin square with 2 x 2 factorial arrangement of treatments. Four lactating Holstein cows were fed a forage-rich diet without LSO (F; F:C = 65:35, dry matter basis), a forage-rich diet with LSO (FO; F:C = 65:32, 3% LSO), a concentrate-rich diet without LSO (C; F:C = 35:65), or a concentrate-rich diet with LSO (CO; F:C = 35:62, 3% LSO). Total-tract digestibility of DM and OM was greater with supplemental LSO. A tendency for greater total-tract digestibility of NDF and ADF also was observed in cows fed LSO. Ruminal digestibility of NDF or ADF decreased when CO was fed compared with C. In contrast, feeding FO increased NDF or ADF digestibility compared with F. Although ruminal starch digestion was nearly complete with all diets, digestibility was greater when cows were fed C or CO compared with F or FO. Bacterial N flow to the duodenum decreased when FO was fed compared with F. In contrast, feeding CO increased bacterial-N flow compared with C. Neither F:C nor LSO supplementation affected ruminal pH or total VFA concentration in ruminal fluid. However, molar proportion of propionate was greater with C or CO compared with F or FO and increased with LSO supplementation regardless of F:C. Molar proportion of n-butyrate decreased with LSO supplementation. Total protozoal numbers in ruminal fluid decreased markedly only when CO was fed. Overall, data show that feeding LSO had no negative effects on total-tract digestion in dairy cows but may decrease ruminal fiber digestibility when fed with high-concentrate diets. The widely spread idea that LSO decreases digestibility, arising from studies with sheep, did not seem to apply to lactating cows fed 3% LSO.

Animals↗
PubMed2005

Digestible reactive lysine in selected milk-based products.

Reactive lysine contents, true ileal reactive lysine digestibility, and true ileal digestible reactive lysine contents were determined in a wide range of processed milk products. A previously validated assay based on determining reactive lysine in both food and ileal digesta, after reaction of these materials with O-methylisourea, was applied. Semisynthetic diets containing milk products as the sole sources of protein and including chromic oxide as an indigestible marker were fed to growing rats. Digesta from the terminal ileum were collected posteuthanasia and, with samples of the diets, analyzed for reactive lysine (homoarginine) contents. True reactive lysine digestibility was determined after correcting for endogenous lysine loss at the terminal ileum of rats fed an enzyme hydrolyzed casein-based diet, followed by ultrafiltration (5000 Da) of the digesta. Digestible total lysine (determined using conventional methods) was also determined. The true ileal reactive lysine digestibility was high (>91%) in all the milk products tested, but was highest in the UHT milk (100%) and lowest in the infant formulas (91 to 93%). Total lysine digestibility (conventional measurement) significantly underestimated reactive lysine digestibility for all the products tested. The mean underestimation ranged from 1.3 to 7.1% units. The mean digestible total lysine content was significantly different from the available lysine content for most of the products examined. In some cases this difference was small (<3%), but for a number of the products (evaporated milk, whole milk protein, lactose hydrolyzed milk powder, and a sports formula) the difference was greater (6.5 to 14%). This would suggest firstly that total lysine and total lysine digestibility determined using conventional methods were inaccurate when applied to some milk-based foods, and secondly that some of the milk products have undergone lysine modification. In general, milk proteins are a highly digestible source of amino acids and lysine.

Amino Acids↗
PubMed1987

Characterization of in situ dry matter and nitrogen digestion of various corn grain forms.

Corn was harvested and stored as either dried shelled, dried ear, high moisture shelled, or high moisture ear. Shelled corns were rolled and ear corns were ground through a forage harvester. High moisture corns were stored in sealed drums. Each corn type was subjected to in situ ruminal digestion with incubation times through 100 h for both original forms and ground at 5 mm. Dry matter, N, and nonprotein DM digestion rates were determined. Unground dried shelled corn possessed the fastest and high moisture ear corn the slowest rate of DM digestion; however, extent of digestion was greatest for high moisture corns within 16 h of ruminal exposure. Grinding increased the rate of shelled corn digestion. Nitrogen digestion rate was fastest for high moisture ear corn and slowest for dried shelled corn. Grinding increased the rate of nitrogen digestion for all corn forms except high moisture shelled corn. Generally, correction for bacterial nitrogen contamination resulted in increased rate of nitrogen digestion. Dried shelled corn possessed the fastest rate of nonprotein DM digestion. Ratio of nitrogen to nonprotein DM rates ranged from .56 to 1.24 for dried shelled corn and high moisture ear corn, respectively. Grinding narrowed this range. Digestion rates of corn forms may not necessarily reflect initial (1 to 16 h) extent of ruminal nutrient availability.

Animal Feed↗
PubMed1989

Nutrient digestion in the large intestine as influenced by forage to concentrate ratio and forage physical form.

Digestion in the large intestine was investigated in four growing steers fitted with duodenal and ileal cannulas. Diets assigned within a 4 x 4 Latin square were: 20% long alfalfa hay and 80% grain; 15% pelleted alfalfa, 5% hay, and 80% grain; 80% hay and 20% grain; and 60% pellets, 20% hay, and 20% grain. Intake of DM was not affected by diet. Organic matter digestion in the large intestine averaged 9, 3, 1, and -4% of total tract digestion for the 20% hay, 20% pellet, 80% hay, and 80% pellet diets, respectively, with significant increases due to high grain diets and long hay. Digestion of CP in the large intestine as a percentage of total tract digestion was unaffected by diet but averaged 3, -1, .3, and -6% for the respective diets. Percentage of total tract starch digestion occurring in the large intestine increased with grain feeding and averaged 6, 3, 1, and 1%, respectively. Digestion of ADF and NDF in the large intestine as a percentage of total tract was unaffected by diet; however, the respective means were 16 and 15, 7 and 7, 5 and 15, and 1 and 1%. In a second trail the same animals were fed a more typical dairy cow diet consisting of 50% grain, 25% corn silage, and 25% long alfalfa hay for a single 18-d period. Digestion in the large intestine accounted for 2.1, .2, 3.6, and 6.4% of total tract digestion of organic matter, starch, ADF, and NDF, respectively; however, feed intake was not as great as in the first trail. Although conditions of these studies differed from those commonly experienced by lactating cows, the underlying principals should apply. The large intestine would be expected to make an even greater contribution to total tract digestion in lactating cows consuming proportionally more DM.

Animal Feed↗
PubMed1994

Influence of corn and sorghum starch on the in vitro kinetics of forage fiber digestion.

In vitro digestion experiments were conducted to examine the interactions of starch source and pH on kinetics and apparent extent of ruminal NDF digestion. Alfalfa hay, bromegrass hay, and each hay with either raw sorghum, raw corn, or pure corn starch in ratios to simulate a 28% NDF diet were incubated at pH 5.5, 6.2, or 6.8. Ash-free NDF was measured at 0, 6, 12, 18, 24, 30, 36, 48, 72, and 96 h of fermentation; logarithmic transformation and linear regression were used to estimate digestion parameters. Significant forage by starch interactions on lag and rate of NDF digestion and significant forage and starch by pH interactions on rate and potential extent of NDF digestion indicated that forage type must be considered when the effect of pH and starch source on NDF digestion is predicted. For both forages, raw or pure corn starch increased lag more than sorghum starch, especially at low pH. Raw sorghum starch decreased the rate of NDF digestion more for bromegrass hay than for alfalfa hay, and pure corn starch had the largest negative impact on fiber digestion, especially at low pH. Sources of starch influenced lag and rate of NDF digestion differently at pH from 6.8 to 5.5, which led to dramatic differences in apparent extent of ruminal NDF digestion.

Animal Feed↗
PubMed1999

Comparison of methods of in vitro dry matter digestibility for ten feeds.

The objectives were to determine if the in vitro dry matter (DM) digestibility was similar by using a traditional method compared to the new DAISYII system and to determine if in vitro DM digestibility was similar for sources of inoculum from two different donor cow diets, all forage or total mixed ration. Ten feeds were digested by the traditional method, the DAISYII method with same feeds in a digestion vessel, and the DAISYII method with different feeds in a digestion vessel. The study used a 2 x 3 factorial arrangement of treatments with two sources of inoculum and three methods. The study was replicated. Source of inoculum affected in vitro DM digestibility; the grass hay donor cow diet resulted in lower digestibility values in six of the 10 feeds tested. Method did not significantly affect the digestibility values, and there were no significant source by method interactions. The DAISYII method can be used to increase labor efficiency in the in vitro DM digestibility analysis, and forages and grains can be analyzed together in a single digestion vessel.

Animal Feed↗
PubMed2000

Effects of pH variation and NaCl on in vitro digestibility of cow's milk proteins in commercially available infant formulas.

Allergenicity and antigenicity of food proteins are generally dependent on their heat-stability and resistance to digestive enzymes. Using the methods of SDS-PAGE and immunoblots, we assessed peptic-digestibility of major milk proteins (casein, beta-lactoglobulin and alpha-lactalbumin) in commercially available infant formulas in acidic pH range (pH 1.5 to 4.0), and we also investigated the effect of NaCl on peptic-digestibility of cow's milk proteins at pH 2.0. The profile of peptic-digestion of casein was similar at pH range from 1.5 to 3.5 where the intact protein of casein rapidly disappeared, whereas slow digestibility of the protein was found at pH 4.0. Beta-lactoglobulin and alpha-lactalbumin were digested at pH range from 1.5 to 2.5 and were almost entirely resistant to peptic digestion at over pH 3.0. In the presence of NaCl at pH 2.0, casein showed a good digestibility by pepsin similar to that in the absence of NaCl, in contrast to beta-lactoglobulin and alpha-lactalbumin both having their peptic-digestibility decreased in the presence of 0.2 M NaCl. As the state of the stomach in new born infants shows low amounts of secretary pepsin and out of optimum pH of peptic activity, low digestibility of beta-lactoglobulin and alpha-lactalbumin in cow's milk based infant formulas at over pH 3.0 is supposed to be responsible for their allergenicity.

Animals↗
PubMed1995

Dietary formulation with cottonseed meal on a total amino acid versus a digestible amino acid basis.

The results of previous experiments indicated that performance of chicks fed corn-cottonseed meal (CSM) diets was inferior to performance of chicks fed a corn-soybean meal (SBM) diet and that true digestibilities of amino acids (AA) in CSM were lower than those in SBM. Therefore, the current study evaluated the formulation of diets containing expander-solvent CSM on an equivalent total AA basis vs an equivalent digestible AA basis compared to a corn-SBM diet. True digestibility of AA in the CSM, corn, and SBM were determined using the precision-feeding assay, with cecectomized roosters and TMEn was determined using conventional roosters. One-week-old male chicks were then fed diets containing corn and SBM only or diets containing increasing levels of CSM from 5 to 40% that were formulated to be equal in total or digestible AA to the corn-SBM diet. All diets contained 20.7% CP and 3,200 kcal TMEn/kg. Weight gain and feed efficiency of chicks fed 20% or more CSM on a total AA basis were less (P < .05) than those of chicks fed the corn-SBM diet. Growth performance of chicks fed up to 20% CSM on a digestible basis was equivalent to that of chicks fed the corn-SBM diet, but dietary levels of 30 or 40% CSM depressed performance even on a digestible basis. Increasing most essential AA by 15% in the 40% CSM (digestible basis) diet had no effect on performance. True digestible AA and ME values determined in chicks were similar to those determined in roosters. These results indicate that formulation of diets containing CSM on a digestible AA basis is superior to formulation on a total AA basis and that at least 20% CSM could be fed to chicks if the diet contains adequate levels of digestible AA and Fe to bind gossypol. The results also indicated that the depression in performance from feeding 30 or 40% CSM diets was not associated with dietary AA levels.

Aging↗
PubMed1996

Bioavailability of the digestible lysine and vilaine in cottonseed and soybean minerals.

Previous research showed that formulation of chick diets containing cottonseed meal (CSM) using true digestible amino acid (AA) values was superior to formulation based on total AA; however, optimum growth performance could not be obtained from high CSM diets even when formulated on a digestible AA basis. Therefore, two experiments were conducted to determine whether the digestible AA in CSM were totally bioavailable or utilizable for protein synthesis. Soybean meal (SBM) was also evaluated for comparison. True digestibility of Lys and Val in CSM and SBM were determined using the precision-fed cecectomized rooster assay. Bioavailability was then assessed by chick growth assay using purified crystalline AA diets deficient in Lys or Val that were supplemented with varying levels of the test AA, CSM,SBM, or AA mixtures simulating the digestible AA composition of CSM and SBM. Multiple regression slope-ratio methodology indicated that bioavailability of the digestible Lys in CSM and the digestible Lys and Val in SBM did not differ (P > 0.05) from 100% for most response parameters. Bioavailability of the digestible Val in CSM ranged from 78 to 96% depending on response parameter. Bioavailabilities of crystalline Lys and Val in the AA mixtures simulating CSM and SBM were generally lower than the bioavailabililties of the digestible Lys and Val in CSM and SBM. The results of this study indicated that the digestible Lys and Val in SBM and CSM as measured by the precision-fed cecectomized rooster assay are totally or almost totally bioavailable for protein synthesis.

Amino Acids↗
PubMedDate not supplied

Survey of public knowledge about digestive health and diseases: implications for health education.

Increasing emphasis in recent years has been placed on health promotion, prevention, and the self-management of health care. These strategies presume the public has sufficient levels of relevant health information, as well as necessary attitudes and skills for the effective use of this information in the management of their own health care. This study tests this assumption as it relates to the level of public knowledge of digestive health and disease, a major health concern affecting an estimated 1 in 10 Americans. This paper reports results of a telephone survey of a representative national sample administered to 1,250 Americans in May 1983 that was designed to assess their level of information about digestive health and disease, comfort in communicating about digestive problems, and preference for health information sources. The results indicate that the American public is largely uninformed and misinformed about digestive health and disease, and they underscore the need for disseminating relevant health information about digestive health and disease to the public to facilitate prevention of digestive health problems and self-management of digestive health care. Health information dissemination is severely complicated by the widespread stigma associated with digestive topics, manifested in the American public's general discomfort in communicating with others about digestive health. These factors necessitate development of sensitive and pervasive digestive health promotion and education programs in the United States.

Attitude to Health↗
PubMedDate not supplied

[The apparent prececal and fecal crude protein digestibility and amino acid absorption of coarse animal feed in fattening pigs].

Digestibility experiments were made with nearly full-grown female fattening pigs supplied with large-scale T-cannulae implanted in the distal ileum to collect the digesta in order to test 11, mainly dried and pelleted, kinds of forage plants (9 samples of lucerne cut at different times as well as green rye and whole plants of maize) as supplements to a basic diet. All samples were analysed to their content of HCl-insoluble ash as marker and their content of amino acids (AA). The calculation of the apparent precaecal and faecal crude protein (CP) digestibility and of the absorption values of the essential AA lysine, methionine, cystine, isoleucine, threonine and tryptophan as well as of glutamic acid was carried out according to the difference method. CP digestibility of the experimental diets was precaecally reduced by 5 to 20 and faecally by 3 to 13 units by supplements of at least 25% of the dry matter; for some of the AA the reduction was even bigger and dispersion between the values of the individual animals was essentially increased in comparison with the basic ration. Faecally, the precaecal differences of CP digestibility and AA absorption caused by the various alfalfa samples, are largely blurred. The CP and AA digestibility of the forage plants partly showed very big individual differences between the animals. Precaecally, a CP digestibility of more than 48% could not be ascertained. Similarly to the lysine content, precaecal lysine absorption from the kinds of roughage showed big differences. Whereas the absorption values for methionine of between 60 and 87% were relatively high, those for cystine were mostly very low, often in the negative range. This underlines that normal digestion processes are strongly influenced by high supplements of forage plants so that the constancy of the digestibility of the basic ration must be doubted under these conditions. The bacterial processes in the large intestine greatly level out the differences found precaecally between the various kinds of feedstuffs. The faecal digestibility and absorption values were considerably above those measured precaecally; methionine digestibility only was mostly considerably diminished in comparison with precaecal values due to synthesis in the large intestine. A significant positive correlation could be calculated between the lysine content of the 9 alfalfa samples (4.0 to 6.6 g/16 g N) and the apparent precaecal values of lysine absorption (2.2 to 67.6%).(ABSTRACT TRUNCATED AT 400 WORDS)

Amino Acids↗
PubMed2001

Investigation of the applicability of a sequential digestion protocol using trypsin and leucine aminopeptidase M for protein identification by matrix-assisted laser desorption/ionization--time of flight mass spectrometry.

An investigation of the applicability of a sequential digestion procedure involving endo- and exoprotease digestion of proteins is reported. The procedure involves the digestion of a protein sample with trypsin, yielding peptide fragments characteristic of the protein. The resulting mixture of peptide fragments is then subjected to N-terminal sequencing with leucine aminopeptidase M (LAP), with matrix-assisted laser desorption ionization-time of flight mass spectrometric analysis of the various digestion products. Several proteins in solution, as well as gel extracted proteins were subjected to this sequential enzyme digestion procedure. The results of these experiments reveal that LAP will preferentially cleave specific peptides of the trypsin digested sample with high efficiency, while leaving other peptides undigested. Also, the length of the amino acid sequence tags that can be generated with this method is limited; the longest sequence tag generated from a single tryptic peptide was four amino acids, even though the digestion was allowed to proceed for long times. In the experiments, N-terminal digestion products were detected as early as two minutes, or as late as 90 minutes, following the addition of LAP to the sample. The method was shown to be effective for subpicomole starting quantities of protein, although with some loss in digestion efficiency at lower concentrations of starting material. This method is useful in providing additional sequence information to increase the level of confidence in protein identification, as illustrated in the identification of bacterial proteins fractionated by high-performance liquid chromatography. In some instances, this method can provide additional sequence information where post source decay and nanospray mass spectrometry failed to generate fragment ion spectra. This is illustrated in an example where the procedure was applied to a membrane protein, CD9, that had been isolated by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Although the sequential digestion procedure requires more human intervention, it is a straightforward method and can be readily implemented.

Cell Line↗
PubMed2000

Microwave digestion of "residual fuel oil" (NIST SRM 1634b) for the determination of trace elements by inductively coupled plasma-mass spectrometry.

A microwave procedure for the digestion of the NIST 1634b reference material "residual fuel oil" in closed pressurized vessels was developed in an attempt to facilitate routine analysis and obtain reproducible conditions or comparable results. The influence of sample size, reagent composition and volume, microwave power, and duration of heating on the digestion procedure was studied. Pressure and temperature inside the reaction vessels were monitored to determine the progression of the reaction and to develop optimal conditions. A nine-step heating program requiring 36.5 min with microwave power not exceeding 450 W in the pulsed mode was found suitable for the digestion of approximately 250 mg fuel oil with a mixture of nitric acid (5.0 mL) and hydrogen peroxide (2.0 mL). The reproducibility of microwave power was determined in terms of the relative standard deviations (n = 3) for temperature (2.7%) and pressure (4.9%) data. The vapor pressures obtained with 5.0 mL Milli-Q water (heated) in an 80-mL digestion vessel showed good agreement with literature data. The excess acid in the resulting digests was removed by evaporation and the concentrations of 24 elements (Ag, Al, As, Ba, Bi, Ca, Cd, Co, Cr, Cu, Fe, Hg, Mg, Mo, Ni, Pb, Sb, Sn, Sr, Ti, Tl, V, U, and Zn) were determined in the diluted digests by inductively coupled plasma mass spectrometry (ICP-MS). The experimental results were in good agreement with the certified and recommended concentrations for eight elements (Al, As, Co, Cr, Ni, Pb, V, Zn) in solutions obtained after one digestion step. An additional digestion step, consisting of intermediate cooling and venting stages, was required for the accurate determination of Fe. No agreement was reached for Ca and Ba even after two-step digestion. The proposed method of digestion provided precise results with relative standard deviations generally less than 5% for most of the elements determined.

Journal Article↗
PubMed1996

Microwave digestion methods for the determination of trace elements in brain and liver samples by inductively coupled plasma mass spectrometry.

Two microwave digestion systems (open-focused and closed-pressurized) were tested for the mineralization of human brain and bovine liver (NIST SRM 1577a) as dissolution steps prior to the determination of 16 trace elements (Bi, Cd, Co, Cs, Cu, Fe, Hg, Mn, Mo, Pb, Rb, Sb, Sn, Sr, Tl, and Zn) by inductively coupled plasma mass spectrometry (ICP-MS). Digestion parameters (mass of sample, digestion mixture, and power/time steps) were optimized using temperature and pressure sensors. Digestions with the open-focused microwave system require larger volumes of conc. HNO(3) and 30% H(2)O(2) than digestions with the closed-pressurized system. Both systems produce correct results for the bovine liver samples. The concentrations obtained for the digests of the open-focused system tend to be less precise than the concentrations from the "closed-pressurized" digests. Because the "open-focused" digests must be diluted to 50 mL to bring the acid concentration to 0.7-2.0 mol/L required by the ICP-MS (closed-pressurized digests need to be diluted to only 20 mL), the detection limits for the system with the open-focused digestion are higher than for the system with the closed-pressurized digestor. The open-focused digestor cannot handle more than 150 mg brain tissue, whereas the closed-pressurized system can mineralize 470 mg. The latter method gave better results with brain tissue than the open-focused system. The preparation of brain tissue as reference material for the determination of trace elements in brain samples is described.

Journal Article↗
PubMed2005

Symbiotic Chlorella sp. of the ciliate Paramecium bursaria do not prevent acidification and lysosomal fusion of host digestive vacuoles during infection.

Each symbiotic Chlorella sp. of the ciliate Paramecium bursaria is enclosed in a perialgal vacuole derived from the host digestive vacuole, and thereby the alga is protected from digestion by lysosomal fusion. Algae-free cells can be reinfected with algae isolated from algae-bearing cells by ingestion into digestive vacuoles. To examine the timing of acidification and lysosomal fusion of the digestive vacuoles and of algal escape from the digestive vacuole, algae-free cells were mixed with isolated algae or yeast cells stained with pH indicator dyes at 25+/-1 degrees C for 1.5 min, washed, chased, and fixed at various time points. Acidification of the vacuoles and digestion of Chlorella sp. began at 0.5 and 2 min after mixing, respectively. All single green Chlorella sp. that had been present in the host cytoplasm before 0.5 h after mixing were digested by 0.5 h. At 1 h after mixing, however, single green algae reappeared in the host cytoplasm, arising from those digestive vacuoles containing both nondigested and partially digested algae, and the percentage of such cells increased to about 40% at 3 h. At 48 h, the single green algae began to multiply by cell division, indicating that these algae had succeeded in establishing endosymbiosis. In contrast to previously published studies, our data show that an alga can successfully escape from the host's digestive vacuole after acidosomal and lysosomal fusion with the vacuole has occurred, in order to produce endosymbiosis.

Animals↗
PubMed1987

Carboxypeptidase Y digestion of band 3, the anion transport protein of human erythrocyte membranes.

The exposure of the carboxyl-terminal of the Band 3 protein of human erythrocyte membranes in intact cells and membrane preparations to proteolytic digestion was determined. Carboxypeptidase Y digestion of purified Band 3 in the presence of non-ionic detergent released amino acids from the carboxyl-terminal of Band 3. The release of amino acids was very pH dependent, digestion being most extensive at pH 3, with limited digestion at pH 6 or above. The 55,000 dalton carboxyl-terminal fragment of Band 3, generated by mild trypsin digestion of ghost membranes, had the same carboxyl-terminal sequence as intact Band 3, based on carboxypeptidase Y digestion. Treatment of intact cells with trypsin or carboxypeptidase Y did not release any amino acids from the carboxyl-terminal of Band 3. In contrast, carboxypeptidase Y readily digested the carboxyl-terminal of Band 3 in ghosts that were stripped of extrinsic membrane proteins by alkali or high salt. This was shown by a decrease in the molecular weight of a carboxyl-terminal fragment of Band 3 after carboxypeptidase Y digestion of stripped ghost membranes. No such decrease was observed after carboxypeptidase Y treatment of intact cells. In addition, Band 3 purified from carboxypeptidase Y-treated stripped ghost membranes had a different carboxyl-terminal sequence from intact Band 3. Cleavage of the carboxyl-terminal of Band 3 was also observed when non-stripped ghosts or inside-out vesicles were treated with carboxypeptidase Y. However, the digestion was less extensive. These results suggest that the carboxyl-terminal of Band 3 may be protected from digestion by its association with extrinsic membrane proteins. We conclude, therefore, that the carboxyl-terminal of Band 3 is located on the cytoplasmic side of the red cell membrane. Since the amino-terminal of Band 3 is also located on the cytoplasmic side of the erythrocyte membrane, the Band 3 polypeptide crosses the membrane an even number of times. A model for the folding of Band 3 in the erythrocyte membrane is presented.

Amino Acid Sequence↗
PubMed1986

Studies of the digestion of bradykinin, Lys-bradykinin, and des-Arg9-bradykinin by angiotensin converting enzyme.

We have studied the degradation of bradykinin, lysyl bradykinin and des-Arg9-bradykinin by the angiotensin converting enzyme. Bradykinin was cleaved at two sites to produce the pentapeptide Arg-Pro-Pro-Gly-Phe plus dipeptides Ser-Pro and Phe-Arg. Lysyl bradykinin was cleaved similarly to release the same dipeptides plus the hexapeptide Lys-Arg-Pro-Pro-Gly-Phe. The tripeptidase activity of ACE was observed when des-Arg9-bradykinin was digested. A single cleavage yielded the above pentapeptide plus Ser-Pro-Phe. Although des-Arg9-bradykinin was the most rapidly digested, when mixtures of des-Arg9-bradykinin and bradykinin or lysyl bradykinin were tested, virtually all of the bradykinin and most of the lysyl bradykinin was digested prior to the onset of digestion of des-Arg9-bradykinin. This was shown to be due to inhibition of des-Arg9-bradykinin cleavage by kinins and kinin-degradation products. The order in terms of potency was bradykinin greater than lysyl bradykinin greater than Ser-Pro much greater than Phe-Arg greater than Arg-Pro-Pro-Gly-Phe. The concentration of chloride ion was an important parameter which affected the rate of digestion of each substrate examined. des-Arg9-bradykinin was not digested by ACE in the absence of sodium chloride and the rate of digestion increased as the chloride concentration was increased to 100-150 mM. On the other hand, increasing NaCl concentration was inhibitory for bradykinin digestion. The rate of Lys-bradykinin digestion was increased from 0 to 1 mM NaCl and decreased thereafter up to physiologic concentration. A half-maximal rate was seen at 100-150 mM NaCl compared to no salt. Of the divalent cations examined, cupric ion inhibited further digestion of des-Arg9-bradykinin at physiologic concentrations. Our data indicate that the rate of degradation of kinins and the nature of the stable final cleavage products in plasma or serum (studied in vitro) are dependent upon the effects of chloride ion, metal ions, and the kinetic effects of multiple metabolites produced by at least two kininases.

Bradykinin↗
PubMed2003

Anaerobic digestion of polyelectrolyte flocculated waste activated sludge.

This work examined how adding one of three polyelectrolyte flocculants (T3052: cationic, T2000: non-ionic, and T1052: anionic) affected the anaerobic digestion of wastewater sludge. Methane production, floc characteristics (morphology and zeta-potential) and process parameters (soluble chemical oxygen demands (SCODs) and reductive potentials) were monitored along the digestion tests. The digestion rates of T2000- and T1052-conditioned sludge resembled that for original sludge. The T3052-flocculated sludge generated methane at a higher rate during the first 6 days of digestion than did the original one. In the following stage, the digestion rate of sludge flocculated with T3052 at dosage exceeding 15 g/kg dried solids declined. For example, at 40 days of digestion the methane production amounts for original, 15 g/kg DS flocculated, and 40 g/kg flocculated sludge were of 136, 105, and 85 g/kg DS, respectively. The role of flocculants could change in different stages of digestion. The dosed polymers had no apparent toxicity to the inoculum used. The changes in SCOD, adenosintriphosphate concentrations, oxidative and reductive potential, and zeta-potentials did not correlate with the noted hindered digestion for T3052-conditioned sludge. Microphotographic observation revealed that the flocs of T3052-conditioned sludge were not only of a large size, but also were resistant to structural deterioration during digestion. Therefore, mass transfer resistance was proposed to account for the hindered digestion efficiency observed for T3052-conditioned sludge.

Anaerobiosis↗