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Bile salt and non-bile salt components in bile affecting micellar cholesterol uptake by rat intestine in vitro.

1. The uptake of micellar cholesterol was measured in sacs of the upper half of everted rat intestine. Sacs of 20 cm length were incubated 1 hr in 25 ml. phosphate buffer containing fatty acid, monoglyceride and (3)H-labelled cholesterol in micellar form with the bile salt, sodium taurocholate, as the dispersing agent.2. Sacs obtained from bile fistula rats (bile duct cannulated 48 hr previously) took up more than twice as much cholesterol as did sacs obtained from untreated control rats.3. In experiments utilizing bile-deficient sacs increasing the sodium taurocholate concentration caused an increase in cholesterol uptake. Conversely, adding a small amount of whole bile caused a decrease in cholesterol uptake.4. The inhibitory effects of the bile were greatly enhanced if the bile was pre-treated with cholestyramine to remove the bile salts.5. It is concluded that bile has a variable effect on intestinal cholesterol absorption depending upon its relative concentration of bile salt and a non-bile salt component having opposite actions.6. It is suggested that the variable effect may be related to the physico-chemical dispersion of cholesterol and that the non-bile salt component may be lecithin.

Animals↗

Vectorial transport of bile salts across MDCK cells expressing both rat Na+-taurocholate cotransporting polypeptide and rat bile salt export pump.

Bile salts are predominantly taken up by hepatocytes via the basolateral Na(+)-taurocholate cotransporting polypeptide (NTCP/SLC10A1) and secreted into the bile by the bile salt export pump (BSEP/ABCB11). In the present study, we transfected rat Ntcp and rat Bsep into polarized Madin-Darby canine kidney cells and characterized the transport properties of these cells for eight bile salts. Immunohistochemical staining demonstrated that Ntcp was expressed at the basolateral domains, whereas Bsep was expressed at the apical domains. Basal-to-apical transport of taurocholate across the monolayer expressing only Ntcp and that coexpressing Ntcp/Bsep was observed, whereas the flux across the monolayer of control and Bsep-expressing cells was symmetrical. Basal-to-apical transport of taurocholate across Ntcp/Bsep-coexpressing monolayers was significantly higher than that across monolayers expressing only Ntcp. Kinetic analysis of this vectorial transport of taurocholate gave an apparent K(m) value of 13.9 +/- 4.7 microM for cells expressing Ntcp alone, which is comparable with 22.2 +/- 4.5 microM for cells expressing both Ntcp and Bsep and V(max) values of 15.8 +/- 4.2 and 60.8 +/- 9.0 pmol.min(-1).mg protein(-1) for Ntcp alone and Ntcp and Bsep-coexpressing cells, respectively. Transcellular transport of cholate, glycocholate, taurochenodeoxycholate, chenodeoxycholate, glycochenodeoxycholate, tauroursodeoxycholate, ursodeoxycholate, and glycoursodeoxycholate, but not that of lithocholate was also observed across the double transfectant. This double-expressing system can be used as a model to clarify vectorial transport of bile salts across hepatocytes under physiological conditions.

Adenoviridae↗

Determination of the faecal excretion of labelled bile salts after i.v. administration of 14C-cholic acid. An evaluation of the bile salt malabsorption before and after surgery in patients with Crohn's disease.

By measuring total faecal radioactivity, correlated to 24-hour enterohepatic circulation, following i.v. administration of 14C-cholic acid, bile salt malabsorption was evaluated before and/or after surgery in 80 patients with Crohn's disease localized to the ileum and/or the colon and the results related to the length of ileum diseased or resected. Before operation bile salt malabsorption was observed only in patients with inflammation of the terminal ileum, but no significant correlation was found between bile salt excretion and the extent of ileal disease. In patients subjected to ileal resection with sacrifice of the ileocaecal valve, bile salt malabsorption correlated strongly to the length of ileum resected. This correlation was about the same in ileostomy patients and in patients subjected to restorative operation. We concluded that determination of 14C in faeces is a more sensitive test than the Schilling test and the faecal fat excretion test in reflecting ileal dysfunction, at least in patients with ileal resections.

Adult↗

Enzymatic sulphation of bile salts in man. Bile salt sulphotransferase activity in percutaneous liver biopsy specimens from patients with liver disease.

Bile salt sulphation in liver disease was investigated by measuring the bile salt sulphotransferase level in percutaneous liver biopsy specimens from 27 patients. The same magnitude of mean specific enzyme activity was found in patients with cholestatic and non-cholestatic liver disease. No significant difference of the mean bile salt sulphotransferase activity was found when patients with and without reduced liver function as evidence from the intravenous galactose tolerance test were compared. The present results indicate that induction of liver bile salt sulphotransferase does not occur to a significant extend in clinical conditions with cholestasis.

Alanine Transaminase↗

Measurement of bile salt hydrolase activity from Lactobacillus acidophilus based on disappearance of conjugated bile salts.

Bile salt hydrolase activity of Lactobacillus acidophilus was measured based on the disappearance of sodium glycocholate and sodium taurocholate from the reaction mixture using HPLC. The amount of sodium glycocholate and sodium taurocholate that disappeared was proportional to the amount of sodium cholate that appeared in the mixture as detected by HPLC. Sodium glycocholate did not precipitate at the enzyme reaction conditions (37 degrees C and pH 5.4) for determining bile salt hydrolase activity. The bile salt hydrolase assay was insensitive to low oxidation-reduction potential when measuring bile salt hydrolase from L. acidophilus, an intestinal microorganism. However, EDTA and freezing temperatures were necessary to maintain stability of the partially purified enzyme during storage.

Amidohydrolases↗

[Introduce Tagsk1 into salt-sensitive callus to improve the capacity of salt-tolerance by micropartical bombardment].

The Tagsk1 (Triticum asetium L. glycogen synthase kinase 1) gene derived from the genome of wheat salt-tolerance mutant RH8706-49 was cloned by PCR. The special primers designed according to full length cDNA sequence of Tagsk1 (AF525086). A binary expression vector pBI121-gsk1 containing Gus and Tagsk1 was constructed. And pBI121-gsk1 was introduced into the callus induced from mature embryos of salt-sensitive wheat H8706-34 and cv. China Spring by particle bombardment. The transformed callus were screened by Kanamycin and 0.5% NaCl. The salt-tolerance callus were obtained, which showed higher ability of salt-tolerance and could diffirentiate roots and buds on the medium containing 0.5% NaCl.

Adaptation, Physiological↗

[Ion exchange interactions on silica gel layers. III. Salts of organic bases with organic acids. Organic acids and sodium salts].

By thin-layer chromatographic and spectroscopic (UV, IR) methods it has been proved that bass and acid are formed from salts of organic bases with organic acids during primary interaction because of the ion exchange behaviour of the silica gel layer. The organic acids formed have interacted with metals on the layer during secondary ion exchange and they migrate further as salts again. It has also been established that the extent of secondary ion exchange between metal silanate groups and acids depends on pKS values of acids. On the basis of investigations it can be ascertained with great probability that organic acids of pKS greater than 7 do not interact with metal silanate groups of silica gel layer. Sodium salts of organic acids of pKS less than 3 values similarly do not interact on the other hand sodium salts of organic acids of pKS greater than 3 values are able to interact with silanol groups. It has been established by flame atomic emission method that salicylic acid interacts with sodium ion and 86% of it migrates further as sodium salicylate.

Carboxylic Acids↗

Electrospray ionization tandem mass spectrometric study of salt cluster ions. Part 1--investigations of alkali metal chloride and sodium salt cluster ions.

Salt cluster ions of alkali metal chlorides ACl (A = Li(+), Na(+), K(+), Rb(+) and Cs(+)) and sodium salts NaB (B = I(-), HCOO(-), CH(3)COO(-), NO(2)(-), and NO(3)(-)), formed by electrospray ionization, were studied systematically by mass spectrometry. The influences on the total positive ion and negative ion currents of variation of solvent, solution concentration, desolvation temperature, solution flow-rate, capillary voltage and cone voltage were investigated. Only cone voltage was found to influence dramatically the distribution of salt cluster ions in the mass spectra observed. Under conditions of normal cone voltage of approximately 70 V, cluster ions having magic numbers of molecules are detected with high relative signal intensity. Under conditions of low cone voltage of approximately 10 V, the distribution of cluster ions detected is characterized by a relatively low average mass/charge ratio due to the presence of multiply charged cluster ions; in addition, there is a marked reduction in cluster ions having a magic number of molecules. Product ion mass spectra obtained by tandem mass spectrometry of cluster ions are characterized by a base peak having a magic number of molecules that is less than and closest to the number of molecules in the precursor ion. Structures have been proposed for some dications and some quadruply charged ions. At pH 3 and 11, the mass spectra of NaCl clusters show the presence of mixed clusters of NaCl with HCl and NaOH, respectively. The effects of ionic radius on 20 distributions of cluster ions for 10 salts were investigated; however, the fine structure of these effects is not readily discerned.

Journal Article↗

Electrospray ionization tandem mass spectrometric study of salt cluster ions: part 2--salts of polyatomic acid groups and of multivalent metals.

Salt cluster ions formed from 0.05 M solutions of CaCl(2), CuCl(2) and Na(A)B (where A = 1 or 2 and B = CO(3)(2-), HCO(3)(-), H(2)PO(4)(-) and HPO(4)(2-)) were studied by electrospray ionization tandem mass spectrometry. The effects on salt cluster ions of droplet pH and of redox reactions induced by electrospray provide information on the electrospray process. CaCl(2) solution yielded salt cluster ions of the form (CaCl(2))(n)(CaCl)(x)(x+) and (CaCl(2))(n)(Cl)(y)(y-), where x, y = 1-3, in positive- and negative-ion modes, respectively. Upon collision induced dissociation (CID), singly charged CaCl(2) cluster ions fragmented, doubly charged cluster ions generated either singly or both singly and doubly charged fragment ions, depending on the cluster mass, and triply charged clusters fragmented predominantly by the loss of charged species. CuCl(2) solution yielded nine series of cluster ions of the form (CuCl(2))(n)(CuCl)(m) plus Cu(+), CuCl(+), or Cl(-). CuCl, the reductive product of CuCl(2), was observed as a neutral component of positively and negatively charged cluster ions. Free electrons were formed in a visible discharge that bridged the gap between the electrospray capillary and the sampling cone brought about the reduction of Cu(2+) to Cu(+). Upon CID, these cluster ions fragmented to lose CuCl(2), CuCl, Cl, and Cl(2). Na(2)CO(3) and NaHCO(3) solutions yielded cluster ions of the form (Na(2)CO(3))(n) plus Na(+) or NaCO(3)(-). Small numbers of NaHCO(3) molecules were found in some cluster ions obtained with the NaHCO(3) solution. For both Na(2)HPO(4) and NaH(2)PO(4) solutions, ions of the form (Na(2)HPO(4))(h), (NaH(2)PO(4))(i), (Na(3)PO(4))(j), (NaPO(3))(k) plus Na(+), PO(3)(-) or H(2)PO(4)(-) were observed. In addition, ions having one or two phosphoric acid (H(3)PO(4)) molecules were observed from the NaH(2)PO(4) solution while ions containing one sodium hydroxide (NaOH) molecule were observed from the Na(2)HPO(4) solution. The cluster ions observed from these four salts of polyatomic acid groups indicate that changes in pH occur in both directions during the electrospray process principally by solvent evaporation; the pH value of the acidic solution became lower and that of the basic solution higher.

Journal Article↗

Noise power spectra and recovery rates obtained with different nebulizer systems in ICP atomic emission spectrometric analyses in the case of different types of salts and salt contents.

The features of different nebulizer systems (cross-flow-, Meinhard- and GMK-system) in the ICP atomic emission spectrometric analyses of solutions with different contents of different salts were evaluated. As basis for the comparison of the nebulizer systems the recovery rates and noise power spectra were used. Both showed that the GMK-system could be used for the widest diversity of salts (sodium chloride, ammonium sulfate and sodium tetraborate) and concentration range (0-10%, (m/v)) followed by the cross-flow-system and the Meinhard-system in ICP-OES using a 1 kW argon ICP. Of all nebulizer systems the nebulizer according to Meinhard has the lowest performance for the salts investigated. The noise power spectra of all nebulizer systems are dominated by interference noise from the sample introduction system. This noise increases with the salt concentration of the solution and the GMK-system shows the lowest increase, followed by the cross-flow- and Meinhard-system.

Journal Article↗

Identification of genes essential for growth at high salt concentrations using salt-sensitive mutants of the cyanobacterium Synechocystis sp. strain PCC 6803.

A collection of 17 salt-sensitive mutants of the cyanobacterium Synechocystis sp. strain PCC 6803 was obtained by random cartridge mutagenesis. The genes coding for proteins essential for growth at high salt concentrations were mapped on the completely known genome sequence of this strain. The two genes coding for enzymes involved in biosynthesis of the osmolyte glucosylglycerol were affected in nine mutants. Two mutants defective in a glycoprotease encoding gene gcp showed a reduced salt resistance. Four genes were identified not previously known to be essential for salt tolerance in cyanobacteria. These genes (slr1799, slr1087, sll1061, and sll1062) code for proteins not yet functionally characterized.

Cyanobacteria↗

A new adenosine subtype-1 receptor antagonist, FK-838, attenuates salt-induced hypertension in Dahl salt-sensitive rats.

We investigated the effects of the adenosine type-1 receptor antagonist FK-838 on salt-induced hypertension in Dahl-Sea salt-sensitive (Dahl S) rats. Dahl S rats fed a high-salt (4% NaCl) diet for 4 weeks were treated with FK-838 or hydrochlorothiazide for 4 weeks and alterations in kidney function and morphologic changes were assessed. FK-838 attenuated the development of hypertension in Dahl S rats, and caused a decrease in aortic weight in a dose dependent fashion. The adenosine antagonist did not produce any detectable metabolic disturbance. The blood pressure reduction by FK-838 was associated with attenuation of glomerular and arterial injury in the kidney. The renal protective effect of FK-838 treatment was associated with a reduction of plasma renin activity and plasma aldosterone concentration. In contrast, the thiazide diuretic, which produced almost the same blood pressure reduction as FK-838, did not attenuate renal damage. These data indicate that adenosine A1 receptor antagonism reduces salt-induced hypertension and the consequent renal injuries.

Animals↗

Reduction of development of left ventricular hypertrophy in salt-loaded Dahl salt-sensitive rats by angiotensin II receptor inhibition.

To determine the effect of the angiotensin II AT1 receptor antagonist losartan (DuP753) on echocardiographic left ventricular (LV) anatomy in Dahl rats on high sodium diet, 27 Dahl salt-sensitive (Dahl-S, 13 on drug and 14 receiving tap water) and 27 Dahl salt-resistant rats (Dahl-R, 13 on drug and 14 receiving tap water) were studied by M-mode echocardiography during 8 weeks of 8% NaCl diet. At the endpoint (after 8 weeks or the last echocardiogram for animals who died earlier), Dahl-S receiving losartan had lower LV mass (1.6 +/- 0.4 g/kg 0.59) than Dahl-S receiving tap water (2.2 +/- 0.7 g/kg 0.59; P < .005), although blood pressure was only partially reduced (167 +/- 29 v 195 +/- 52; P = .05). This difference was mainly due to lower LV wall thickness (P < .02), with a less consistent decrease in LV chamber size in Dahl-S receiving losartan. Blood pressure was normal in Dahl-R (tap water group = 116 +/- 11 mm Hg; losartan group = 115 +/- 13 mm Hg) and losartan had no effect on LV mass (1.6 +/- 0.4 g/kg 0.59) in both groups). In the majority of rats, echocardiographic measurements were compared between the end of second or third week and the last available study: LV mass increased in salt-loaded Dahl-S receiving tap water (1.6+/- 0.6 to 2.1 +/- 0.7 g/kg 0.59, P < .04) and was stable in Dahl-S receiving losartan (1.5 +/- 0.1 to 1.5 +/- 0.3 g/kg 0.59), paralleling changes in LV chamber dimension. Thus, a high salt diet leads to hypertension and eccentric LV hypertrophy in Dahl-S but not in Dahl-R. Inhibition of angiotensin II AT1 receptors reduces the development of LV hypertrophy in Dahl-S rats despite lack of efficient control of blood pressure.

Analysis of Variance↗

Difference in water intake but not in renal sympathetic nerve activity in response to central salt-loading or angiotensin II in awake Dahl salt-sensitive and -resistant rats.

Experiments were conducted to examine whether renal sympathetic nerve activity (RSNA) and water intake in response to central salt-loading or angiotensin II (A II) differ between freely-moving Dahl salt-sensitive (DS) and -resistant (DR) rats maintained on a low-salt diet. Intracerebroventricular (i.c.v.) administration of hypertonic saline (0.3 M, 1 microl/min, 20 min) or A II (100 ng/1 microl) evoked water intake, pressor response and suppression of RSNA in both strains. The cumulative water intake in DS rats over a 60-min period after i.c.v. infusion of hypertonic saline or A II was significantly attenuated compared with that in DR rats. The RSNA response did not show a significant difference between the strains. These results demonstrate that water intake, but not RSNA response to acute central salt-loading or A II differ between awake DR and DS rats.

Angiotensin II↗

Dahl salt-sensitive and salt-resistant rats: examination of learning and memory performance, blood pressure, and the expression of central nicotinic acetylcholine receptors.

Substantial human and animal data suggest a correlation between hypertension and memory impairment that may appear prior to overt manifestations of cerebrovascular pathology. It is unclear, however, whether hypertension plays a causal role in these memory deficits, whether hypertension and cognitive impairment are each based in family history and not interdependent, or whether a combination of these factors is important. The purpose of this study was to assess whether deficits in memory performance and nicotinic acetylcholine receptors were present in Dahl salt-sensitive rats (as observed previously in spontaneously hypertensive rats) and whether the presence of hypertension per se (induced with an 8% Na(+) diet) contributed to the deficits. Memory was assessed in a passive avoidance task, an eight-arm radial arm maze and in a water maze task, and nicotinic receptors were measured via quantitative receptor autoradiography utilizing [125I]alpha-bungarotoxin and [3H]epibatidine. Salt-sensitive rats exhibited impaired performance in both spatial learning tasks, but not the passive avoidance task, compared to controls (salt-resistant strain) and they exhibited reductions in nicotinic receptors labeled by [125I]alpha-bungarotoxin but not [3H]epibatidine in some brain regions, including some areas important for memory (e.g. the hippocampus and amygdala). In the radial arm maze, the degree of memory impairment and in binding studies the reduced expression of nicotinic receptors each failed to correlate with the highest blood pressures, and the salt-sensitive animals were impaired relative to controls whether or not the high Na(+) diet was administered. In contrast, higher blood pressures did correlate with inferior task performance in the water maze. These findings may suggest that the genetics of the subjects were critical for performance when appetitive drives were involved, but diet (and perhaps hypertension) were key to performance when memory did not involve appetitive drives or mechanisms. Overall, the data obtained from Dahl rats appear to support the role of family history (selective breeding in rats) as underlying the reductions in central nicotinic acetylcholine receptors, whereas both family history and hypertension may contribute to poor cognitive performance.

Animals↗

Optical sensors and the salt effect: a dual-transducer approach to acidity determination in a salt-containing concentrated strong acid.

A dual-transducer approach has been developed to decompose the optical signals of acid sensors in salt-containing concentrated acid solutions and to give acid and salt concentrations in concentrated LiCl-HCl, CaCl2-HCl, and AICl3-HCl solutions, respectively. The optical acid sensors in this approach are films of porous sol-gel SiO2 or SiO2-Nafion composite doped with low-pKa indicators. A novel linear relationship (dA/dCsalt)cCacid = beta x (dA0/dCacid)Csalt = 0 (A = absorbance of the sensor in a salt-containing HCl solution; A0 = absorbance of the sensor in a salt-free acid solution) was found, and the current approach is based on a set of nonlinear equations derived from this relationship.

Journal Article↗

Association products and conformations of salt-dissociated and acid-extracted histones. A two-phase procedure for isolating salt-dissociated histones.

We present an extremely rapid and efficient method for the separation of salt-dissociated histones from DNA in which the macromolecular components of chicken erythrocyte chromatin are partitioned in a two-phase system of the water-soluble, nonionic polymers, poly(ethylene glycol) and dextran. We have compared the association products and conformations of salt-dissociated histones purified with the two-phase procedure and histones that had been extracted with 0.4 M H2SO4. In the gel chromatography system of D. R. vander Westhuyzen and C. von Holt (1971), FEBS Lett. 14, 333-337] the association products of salt-dissociated and acid-extracted histones are indistinguishable. Furthermore, the circular dichroism spectra of histones prepared with the two methods are identical within experimental error. These results indicate that histones extracted, with sulfuric acid can adopt conformations at least very similar to those of salt-dissociated preperties of total erythrocyte histones are the same in 2 M NaCl as those of these histones bound to DNA in chromatin in 1 mM Tris-Cl (pH 7.5). This result and the studies of Weintraub et al. [Weintraub, H., Palter, K., and Van Lente, F. (1975), Cell 6, 68-110] on the patterns of tryptic digest products of histones strongly suggest that in 2 M NaCl the histones exist in conformations very similar to their conformations when bound to DNA. The concept of native histone conformations is discussed in light of our results.

Animals↗

Electron-transfer salts of 1,2,3,4,5-pentamethylferrocene, Fe(II)(C5Me5)(C5H5). Structure and magnetic properties of two 1:1 and two 2:3 Fe(C5Me5)(C5H5) electron-transfer salts of tetracyanoethylene.

The reaction of Fe(II)(C5Me5)(C5H5), FeCpCp, with percyano acceptors, A [A = C4(CN)6 (hexacyanobutadiene), TCNQF4 (perfluoro-7,7,8,8-tetracyano-p-quinodimethane), and DDQ (2,3-dichloro-5,6-dicyanobenzoquinone)], results in formation of 1:1 charge-transfer salts of [Fe(III)CpCp]*]*+[A]*- composition. With A = TCNQ (7,7,8,8-tetracyano-p-quinodimethane) a 1:2 electron-transfer salt with FeCpCp forms. With A = TCNE (tetracyanoethylene) a pair of 1:1 salts as well as a pair of 2:3 salts of [FeCpCp]2[TCNE]3.S (S = CH2Cl2, THF) have been isolated and characterized by single-crystal X-ray diffraction. [FeCpCp][TCNE] consists of parallel 1-D.D(*+)A(*-)D(*+)A(*-)D(*+)A(*-). chains, while [FeCpCp][TCNE].MeCN has a herringbone array of D(*+)A2(2-)D(*+) dimers separated by solvent molecules. Although each [TCNE](-) is disordered, the diamagnetic [TCNE]2(2-) dimer is structurally different from those observed earlier with an intradimer separation of 2.79 A. The [TCNE](-) in the 2:3 [FeCpCp]2[TCNE]3.S exists as an eclipsed diamagnetic [TCNE]2(2-) dimer with an intradimer ethylene C.C separation of 2.833 and 2.903 A for the CH2Cl2- and THF-containing materials, respectively. The bond distances and angles for all the cations are essentially equivalent, and the distances are essentially equivalent to those previously reported for [FeCp2](*+) and [FeCp2](*+) cations. The average Fe-C5H5-ring and Fe-C5Me5-ring centroid distances are 1.71 and 1.69 A, respectively, which are 0.05 A longer than reported for Fe(II)CpCp. The one-electron reduction potential for Fe(II)CpCp is 0.11 V (vs SCE). The 5 K EPR of [FeCpCp](*+)[BF4](-) exhibits an axially symmetric powder pattern with g(parallel) = 4.36 and g(perpendicular) = 1.24, and the EPR parameters are essentially identical to those reported for ferrocenium and decamethylferrocenium. The high-temperature magnetic susceptibility for polycrystalline samples of these complexes can be fit by the Curie-Weiss law, chi = C/(T - theta), with low theta values and mu(eff) values from 2.08 to 3.43 mu(B), suggesting that the polycrystalline samples measured had varying degrees of orientation. [FeCpCp][TCNE] exhibits the highest effective moment of 3.43 mu(B)/Fe and weak ferromagnetic coupling, as evidenced from the theta of 3.3 K; however, unexpectedly, it does not magnetically order above 2 K. The formation of the four phases comprising FeCpCp and TCNE emphasizes the diversity of materials that may form and the present inability to predict neither solid-state compositions nor structure types.

Journal Article↗