Search PubMed⌕ Search

Biomedical subjects

A Jonas

Publications and source records attributed to A Jonas.

At least 55 records · Page 3Linked to original sources
PubMed1996

Colchicine enhances intestinal permeability in patients with familial Mediterranean fever.

OBJECTIVE: Colchicine therapy is complicated by frequent gastrointestinal adverse effects. METHODS: We compared intestinal permeability in 21 patients with familial Mediterranean fever on long-standing colchicine therapy (mean 5.8 years) and significant gastrointestinal complaints and 12 untreated patients and 14 healthy volunteers. The double probe (lactulose/mannitol) permeability test was performed using a hyperosmolar test solution (1580 mosmol) and the differential urinary recovery ratios were calculated. RESULTS: Familial Mediterranean fever patients on colchicine therapy had significantly higher lactulose/mannitol urinary excretion ratios (0.073) compared to untreated patients (0.035) and to healthy controls (0.021). Untreated familial Mediterranean fever patients had significantly greater urinary lactulose/mannitol recovery ratios than controls (P < 0.02). No correlation was found between the degree of enhanced permeability and the length of exposure to the drug or the severity of clinical symptoms. CONCLUSIONS: Intestinal permeability was significantly enhanced in patients with familial Mediterranean fever treated with colchicine.

Adolescent↗
PubMed1996

Serum pepsinogen I in childhood Helicobacter pylori gastritis: its relation to mucosal peptic activity.

Serum pepsinogen I (PG I) levels are raised in children with Helicobacter pylori gastritis. To ascertain if this is due to increased production or to increased secretion of pepsin by chief cells, we measured mucosal peptic activity in antrum and gastric body mucosal homogenates and correlated it to serum PG I levels in 122 children with and without H. pylori gastritis. In patients infected with H. pylori, mucosal peptic activity was decreased when compared to control and to children with non H. pylori gastritis. Serum PG I levels were increased (P < 0.001) and were inversely related to mucosal peptic activity (P < 0.01). These findings suggest that H. pylori can promote a leakage of pepsinogen into the circulation.

Adolescent↗
PubMed1996

High yield overexpression and characterization of human recombinant proapolipoprotein A-I.

Human apolipoprotein A-I (apoA-I) is the major protein component of high density lipoproteins (HDL) where it defines the particle structure and stability and functions as the main activator of the enzyme lecithin:cholesterol acyltransferase (LCAT). ApoA-I is expressed in the liver as a preproprotein that is targeted to the endoplasmic reticulum for secretion; in plasma, an unknown protease removes the six amino acid long propeptide. In this study, the cDNA coding the human proapoA-I was cloned into an Escherichia coli vector; the overexpressed protein was purified to 99% homogeneity and was extensively characterized together with mature apoA-I purified from plasma. SDS-PAGE, mass spectrometry, and Edman sequence analysis showed that the initial Met residue needed for translation in E. coli is posttranslationally removed from the N-terminal sequence of the proapoA-I. The structural and functional analyses were carried out on the lipid-free and the lipid-bound proteins. ProapoA-I self associated, interacted with dimyristoyl phosphatidylcholine vesicles, and formed secondary structures very similar to the lipid-free apoA-I. Reconstituted HDL particles made with two initial molar ratios of palmitoyloleoyl phosphatidylcholine/cholesterol/apolipoprotein/Na-cholate had identical particle sizes and distributions when apoA-I or proapoA-I were used. Particles having diameters of 79 A and 98 A, containing two apoA-I or proapoA-I molecules per particle, were isolated and characterized. The particles contained the same amounts of alpha-helical structure, had very similar fluorescence properties, and activated LCAT equally well. We conclude that proapoA-I expressed and purified from E. coli is functionally and structurally indistinguishable from mature apoA-I purified from plasma when analyzed in vitro. Therefore, this recombinant proapoA-I and mutants derived from it will be important sources of protein for analyzing apoA-I structure and function, as well as for studies of proapoA-I processing.

Apolipoprotein A-I↗
PubMed1995

High-pressure proton NMR study of lateral self-diffusion of phosphatidylcholines in sonicated unilamellar vesicles.

Effects of pressure on the lateral diffusion of phospholipid molecules in sonicated pure 1,2-dipalmitoylphosphatidylcholine (DPPC) and 1-palmitoyl-2-oleoylphosphatidylcholine (POPC) vesicles (15 wt%) in D2O were examined using the high-pressure proton NMR rotating frame spin-lattice relaxation time (T1rho) method. Proton T1rho were measured at pressures from 1 bar to 5000 bar and at temperatures of 50 degrees C to 70 degrees C for DPPC and 5 degrees C to 35 degrees C for POPC. The T(-1)1rho values were plotted as a function of the square root of the spin-locking field angular frequency (omega1(1/2) and the lateral diffusion coefficient (D) calculated from the slope. Pressure effects on lateral diffusion were observed in the liquid-crystalline (LC) phase. The lateral diffusion coefficient exhibited sharp decreases in response to the various pressure-induced phase transitions encountered. However, pressure had little, if any, effect on lateral diffusion in the pressure-induced gel I (GI) phase and pressure-induced interdigitated gel (Gi) phase. The activation volumes for diffusion were calculated from the slopes from plots of In D versus pressure for both DPPC (37 ml/mol at 50 degrees C, 34 ml/mol at 60 degrees C and 25 ml/mol at 70 degrees C) and POPC (16 ml/mol at 5 degrees C, 9 ml/mol at 20 degrees C and 6 ml/mol at 35 degrees C) sonicated vesicles in the LC phase. The activation energy for diffusion (Ea) was calculated using the slopes from plots of In D versus the inverse of the temperature (1/T) for both DPPC and POPC in the LC phase (3.5 kcal/mol and 3.9 kcal/mol, respectively) and for both DPPC and POPC in the GI phase (6.0 kcal/mol and 4.4 kcal/mol, respectively). From the lateral diffusion coefficient and line width data pressure-temperature phase diagrams for sonicated pure DPPC and POPC vesicles were constructed. The values of the temperature to pressure equivalence of DPPC (dTm/dP) were estimated to be 22.1 degrees C/kbar for the LC to GI phase transition and 28.6 degrees C/kbar for the GI to Gi phase transition. The value of the temperature to pressure equivalence of POPC for the LC to GI phase transition was estimated to be 19.0 degrees C/kbar.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1995

NMR study of the cold, heat, and pressure unfolding of ribonuclease A.

The reversible cold, heat, and pressure unfolding of RNase A and RNase A--inhibitor complex were studied by 1D and 2D 1H NMR spectroscopy. The reversible pressure denaturation experiments in the pressure range from 1 bar to 5 kbar were carried out at pH 2.0 and 10 degrees C. The cold denaturation was carried out at 3 kbar, where the protein solution can be cooled down to -25 degrees C without freezing. Including heat denaturation experiments, the experimental data obtained allowed us to construct the pressure--temperature phase diagram of RNase A. The experimental results suggest the possibility that all three denaturation processes (cold, heat, and pressure) lead to non-cooperative unfolding. The appearance of a new histidine resonance in the cold-denatured and pressure-denatured RNase A spectra, compared to the absence of this resonance in the heat-denatured state, indicates that the pressure-denatured and cold-denatured states may contain partially folded structures that are similar to that of the early folding intermediate found in the temperature-jump experiment reported by Blum et al. [Blum, A. D., et al. (1978) J. Mol. Biol. 118, 305]. A hydrogen-exchange experiment was performed to confirm the presence of partially folded structures in the pressure-denatured state. Stable hydrogen-bonded structures protecting the backbone amide hydrogens from solvent exchange were observed in the pressure-denatured state. These experimental results suggest that the pressure-denatured RNase A displays the characteristics of a the inhibitor 3'-UMP show that the RNase A-inhibitor complex is more stable than RNase without the inhibitor.

Animals↗
PubMed1995

Cholesterol transfer from low density lipoproteins to reconstituted high density lipoproteins is determined by the properties and concentrations of both particles.

Cholesterol spontaneously transfers from low density lipoproteins (LDL) to high density lipoproteins (HDL). This transfer is important physiologically as it supplies the major portion of cholesterol for the lecithin:cholesterol acyltransferase reaction and is one mechanism for the reduction of atherogenic LDL cholesterol. The objective of this work was to examine the properties of both HDL and LDL which modulate cholesterol transfer, as well as to obtain the relevant kinetic constants for the transfer at concentrations of lipoproteins approaching those existing in vivo. To examine the effects of HDL structural parameters on cholesterol transfer, we prepared reconstituted HDL particles with saturated or unsaturated phospholipid, with apolipoprotein AI or apolipoprotein AII, with increasing size and phospholipid content, and with increasing initial contents of cholesterol. We also prepared five LDL subfractions of variable density and size. The kinetics of cholesterol mass transfer were measured by incubating LDL with rHDL at 37 degrees C, separating the lipoproteins by dextran sulfate/Mg2+ precipitation of LDL at timed intervals, and analyzing rHDL cholesterol content. The cholesterol content of rHDL at equilibrium, Ceq, and the half-time for transfer, t1/2, as well as the ratio of the lipid surface areas of LDL to rHDL were used in the analysis of the kinetic data by the aqueous diffusion model for lipid transfer developed by Nichols and Pagano [(1982) Biochemistry 21, 1720-1726]. The only variables that significantly affect the C(eq) and/or t1/2 are the phospholipid content and composition of the rHDL and the size or density of the LDL particles.(ABSTRACT TRUNCATED AT 250 WORDS)

Cholesterol↗
PubMed1995

Properties of an N-terminal proteolytic fragment of apolipoprotein AI in solution and in reconstituted high density lipoproteins.

Limited proteolysis was used to study the domain structure and to produce a large N-terminal fragment of human apolipoprotein AI (apoAI). Digestion of reconstituted high density lipoprotein (rHDL) prepared with apoAI and dipalmitoyl phosphatidylcholine or palmitoyloleoyl phosphatidylcholine by chymotrypsin, trypsin, elastase, and subtilisin generated a major fragment of 22 kDa. Under milder conditions proteolysis of lipid-free apoAI produced a fragment of similar size. The fragments shared the same N terminus as intact apoAI, and the chymotryptic fragment had a molecular weight of 22,384 as determined by electrospray ionization mass spectrometry. Thus the fragment consists of the N-terminal 192 amino acid residues of apoAI, and the region around Tyr192 seems to be especially accessible to proteases. In aqueous solution the fragment, apoAI-(1-192), had an alpha-helix content similar to that of apoAI (approximately 52%) but existed only as monomers and dimers. ApoAI-(1-192) lysed dimyristoyl phosphatidylcholine liposomes slowly compared with apoAI but did form rHDL complexes with palmitoyloleoyl phosphatidylcholine or dipalmitoyl phosphatidylcholine when prepared by the sodium cholate dialysis method. ApoAI-(1-192) rHDL exhibited sizes and size distributions distinct from apoAI rHDL but displayed similar stability against denaturation. The isolated apoAI-(1-192) rHDLs retained a high ability to activate lecithin-cholesterol acyltransferase, comparable with the most effective apoAI rHDL. The results suggest that the C-terminal domain of apoAI is crucial for self-association and initial lipid binding but is not involved in specific lecithin-cholesterol acyltransferase activation.

Amino Acid Sequence↗
PubMed1995

One and two dimensional 1H-NMR studies of pressure and tetracaine effects on sonicated phospholipid vesicles.

One and two dimensional 1H-NMR experiments have been performed to study the molecular order and dynamics of 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) unilamellar vesicles under the influence of high pressure and the local anesthetic, tetracaine (TTC), which can also be considered as a model-charged amphiphile. The TTC molecules have an ordering effect on the headgroups but a disordering effect on the acyl chains, whereas, pressure has an ordering effect on the acyl chains. The results from 2D NOESY experiments on pure DPPC vesicles show that the intensities of NMe3/CH3, NMe3/(CH2)n, and CH3/(CH2)n cross-peaks increase with increasing pressure in the liquid-crystalline phase but decrease with pressure in the gel phase, further suggesting that the appearance of the cross-peaks between the two extremes of the DPPC molecules is due to spin-diffusion. For the DPPC/TTC vesicles, the 2D NOESY experiments confirm that charged TTC molecules are located in the headgroup region of the DPPC bilayers and, in the gel phase, also suggest the formation of an interdigitated gel phase.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1995

High pressure 2H-NMR study of the order and dynamics of selectively deuterated dipalmitoyl phosphatidylcholine in multilamellar aqueous dispersions.

High pressure 2H multipulse NMR techniques were used to investigate the effects of pressure on the structure and dynamics of selectively deuterated 1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine (DPPC) multilamellar aqueous dispersions. The samples were deuterated on both chains at positions 2, 9, or 13. The deuterium lineshapes, the spin-lattice relaxation times, T1, and the spin-spin relaxation times, T2, were measured as a function of pressure from 1 bar to 5 kbar at 50 degrees C for the three deuterated DPPC samples. This pressure range permitted us to explore the phase behavior of DPPC from the liquid-crystalline (LC) phase through various gel phases such as the Gel I (P beta), Gel II (L beta), Gel III, Gel X, and the interdigitated, Gel i, gel phase. Pressure had an ordering effect on all chain segments both in the LC phase and various high pressure gel phases as indicated by the increase in SCD bond order parameter and the first moment, M1, with pressure. Compared with the adjacent gel phases, the Gel i phase had the highest order. Also, in all gel phases the carbon-9 segment of the chains had the most restricted motions in contrast to the LC phase, where the carbon-2 segment was the most restricted. In the LC phase, T1 and T2 values for all segments decreased with pressure, indicative of the fast correlation time regime. Similarly, T1 decreased with pressure in the Gel I and the interdigitated Gel i gel phases but changed to the slow correlation time regime at the Gel i/Gel II phase transition. For T2, which reflects slow motions, the transition to the slow correlation time regime occurred already at LC/Gel I phase transition. Considering the various motions which contribute to relaxation, the behavior of T1 and T2 in the Gel 11 through Gel X phases showing discontinuities and slope changes at the phase transitions was, as expected, quite complex.In addition we found a straight line relationship for T-1 vs. S2D, and T-1 vs. S2CD for the deuterons in the 9 and 13 positions in the LC phase in the pressure range investigated.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1995

Colchicine induces enhanced intestinal permeability in the rat.

Intestinal permeability was determined in rats receiving colchicine 0.5 +/- 0.15 mg day-1 in drinking water (30 mg L-1) for periods up to 23 days. The lactulose/mannitol method was used to determine whole gut permeability before and on days 2, 4, 8, 18 and 23 of colchicine administration. The 8-h urinary lactulose excretion following the test meal increased significantly in rats receiving colchicine, compared with the pretreatment value. Increased lactulose permeability was present after 2 days and remained stable throughout the experimental period. Mannitol urinary excretion was not changed. Colchicine increases intestinal tight junction permeability by an as yet undetermined mechanism.

Animals↗
PubMed1995

Colchicine-induced lactose malabsorption in patients with familial Mediterranean fever.

Abdominal pain and diarrhea are frequent side effects of chronic colchicine therapy. Drug-induced lactose deficiency has been demonstrated in the experimental animal. Lactose malabsorption was assessed by the lactose breath test in 23 patients with familial Mediterranean fever (FMF) receiving colchicine for 0.25-15 years (mean 3.16). Twenty FMF patients not receiving colchicine and 38 non-FMF lactose malabsorbers served as controls. Patients receiving colchicine had a significantly higher percentage of lactose malabsorption (20/23, 87%) versus nontreated FMF patients (13/20, 65%; P < 0.05). Lactose intolerance was also more prevalent in colchicine-treated patients (17/23, 74%) versus nontreated FMF (5/20, 25%; P < 0.0005) and control lactose malabsorbers (16/38, 42%; P < 0.01). Of the 12 patients investigated before and 3 months after colchicine administration, 7 showed induction or aggravation of lactose malabsorption. The lactose-free diet resulted in partial improvement of symptoms. Colchicine induces significant lactose malabsorption in FMF patients and this is partially responsible for the gastrointestinal side effects of the drug.

Abdominal Pain↗
PubMed1994

Native-like structure and self-association behavior of apolipoprotein A-I in a water/n-propanol solution.

The effect of n-propanol on the secondary and tertiary structure of human apolipoprotein A-I (apoA-I), an interfacial protein, was investigated using near and far ultraviolet (UV)-circular dichroism (CD) and fluorescence spectroscopy, as well as limited proteolytic digestion with trypsin, and cross-linking with bis(sulfosuccinimidyl) suberate. The structure of apoA-I in n-propanol (30%, v/v) was compared with that in Tris buffer and in reconstituted, spherical or discoidal, high density lipoproteins (rHDL). Addition of n-propanol to apoA-I in Tris buffer induces major changes in its near and far CD spectra: alpha-helical content increases by 27% and the near UV-CD spectrum becomes very similar to that of apoA-I in rHDL particles. Fluorescence spectral, lifetime, and polarization results, and quenching by KI confirm that major structural changes occur in the N-terminal half of apoA-I as n-propanol is added: the Trp residues become more exposed to solvent than in buffer alone or in rHDL. Higher concentrations of guanidine hydrochloride or urea are required to denature apoA-I in n-propanol than in buffer alone, but a similar free energy of unfolding is observed. The N-terminus of apoA-I is relatively resistant to trypsin digestion and the C-terminus has equivalent digestion sites for apoA-I in the three states, but the kinetics of digestion are much slower in n-propanol and in rHDL compared to apoA-I in Tris buffer. Cross-linking experiments reveal that dimers of apoA-I exist in n-propanol, in contrast to dimers plus multimeric aggregates in Tris buffer. From these results we conclude that in 30% n-propanol the structure of apoA-I approaches that of 'native' lipid-bound apoA-I, in contrast to its structure in the aqueous Tris buffer.

1-Propanol↗
PubMed1994

Binding of lecithin:cholesterol acyltransferase to reconstituted high density lipoproteins is affected by their lipid but not apolipoprotein composition.

The reaction of lecithin:cholesterol acyltransferase (LCAT) with high density lipoproteins (HDL) is of critical importance in reverse cholesterol transport. We studied the relationship between LCAT binding and HDL composition using two assays to determine LCAT binding affinity for discoidal reconstituted HDL (rHDL). We prepared rHDL with egg phosphatidylcholine (egg-PC), cholesterol, and either apolipoprotein (apo) A-I or apoA-II. The rHDL, identical in lipid content, had sizes of 96 and 100 A, respectively. Binding constants (Kd) determined by the activity-inhibition method were 2.2 +/- 0.3 x 10(-7) and 1.1 +/- 0.3 x 10(-16) M, whereas those determined by a solid-phase method were 3.3 +/- 0.9 x 10(-7) and 3.7 +/- 0.9 x 10(-7) M for apoA-I and apoA-II rHDL, respectively. The stoichiometry was 1 LCAT bound/rHDL. The appVmax/appKm for apoA-I was 80-fold higher than for apoA-II rHDL. The large difference between LCAT-binding constants and enzymatic activity measurements for the two particles suggests that LCAT binding and activation by apolipoproteins are independent events. To determine the effects of phospholipid headgroup on LCAT binding affinity, we tested rHDL containing up to 16 mol % of egg phosphatidylethanolamine (egg-PE), egg phosphatidic acid (egg-PA), or bovine phosphatidylserine (PS). Kd was unchanged for PS rHDL, but increased 6-fold with increasing PE content. AppVmax increased with PE content, but decreased with PS or PA relative to egg-PC controls. AppKm increased in PE rHDL, but remained unchanged in rHDL with PA or PS. Fluorescence characterization of the lipid domains of rHDL shows small differences in the polarity of the headgroup region of PE rHDL. Thus, LCAT binding is influenced by the lipid, but not the protein composition of rHDL. AppVmax values reflect active site preferences, while appKm values reflect interfacial binding affinity.

Apolipoproteins↗
PubMed1994

Acute acalculous cholecystitis in an infant after cardiac surgery.

Acute acalculous cholecystitis is a virulent disease that occurs most commonly in critically ill patients. Imaging studies may facilitate earlier diagnosis and may decrease the morbidity and mortality rates. The authors report on a 30-month-old child with acute acalculous cholecystitis after surgical repair of tetralogy of Fallot. A review of the clinical pathogenesis, diagnosis, and treatment is included.

Acute Disease↗
PubMed1994

Transfer of cholesterol from Ob1771 cells or LDL to reconstituted, defined high density lipoproteins.

We used defined, reconstituted high density lipoproteins (rHDL) to study the effects of structure and composition of these particles on their role as cholesterol acceptors from cell membranes or from low density lipoproteins (LDL). Three discoidal rHDL and one spherical rHDL with distinct apolipoprotein A-I conformations, diameters and compositions were used in conjunction with Ob1771 cells to measure the rate of [3H]cholesterol efflux from the cells, direct binding to the cells, and competition with native HDL3 for binding. In addition, the same rHDL particles were used to study the kinetics of cholesterol mass transfer from LDL. The results show that the rates of cholesterol transfer depend on the nature of the donor (t1/2 11-19 min from LDL, and t1/2 5 h from the cells), on the phosphatidylcholine/cholesterol ratio in the acceptors (the closer this ratio is to the equilibrium value, the slower is the rate), and on the diameter of the acceptors (the smallest particles have the lowest t1/2 for cholesterol uptake from LDL, and are the most effective acceptors of [3H]cholesterol from cells after their phospholipid content is taken into account). The cholesterol uptake by the rHDL, both from the cells and from LDL, is determined mostly by the phospholipid pool available in the acceptors. Binding to the cells was equivalent for all the rHDL (Kd = 38-67 micrograms/ml) and comparable to HDL3, suggesting that the differences in apoA-I conformation have no effect on the binding to cells. Finally we observed that exposure of rHDL to cells may lead to remodeling of some of the lipoprotein particles.

Adipocytes↗
PubMed1993

Bordetella bronchiseptica pneumonia in a koala.

A 3-year-old male koala (Phascolarctos cinereus) was evaluated because of severe acute respiratory distress. Despite medical treatment, the koala died. Gross and histologic lesions consistent with acute bacterial pneumonia were identified at necropsy and Bordetella bronchiseptica was isolated from pulmonary tissue. There are 2 prior reports from Australia of fatal B bronchiseptica pneumonia in captive koalas.

Acute Disease↗
PubMed1993

Structural properties of high density lipoprotein subclasses homogeneous in protein composition and size.

We isolated native high density lipoprotein (HDL) subclasses homogeneous in size and in their protein content with the objective of investigating the differences and similarities in their apolipoprotein AI (apoA-I) structures. Defined particles were isolated from ultracentrifugally prepared HDL by immunoaffinity and gel-filtration chromatography. The isolated 88-A LpAI, 106-A LpAI, 96-A LpAI/AII particles (LpAI, particles contain only apoA-I; LpAI/AII, particles contain apoA-I and apoA-II), together with a 93-A reconstituted HDL were analyzed for purity, composition, and content of apolipoprotein molecules per particle, and were examined by far and near circular dichroism and intrinsic fluorescence spectroscopic methods, as well as by reaction kinetics with lecithin:cholesterol acyltransferase. The spectroscopic analyses indicated that the secondary structures and three-dimensional arrangements of apoA-I in all these particles are remarkably similar: their tryptophan residues are located in similar nonpolar environments and become exposed to increasing concentrations of guanidine hydrochloride in comparable denaturation steps; the 60-65% alpha-helical structures in apoA-I are denatured in similar patterns with 0-5 M denaturant concentrations. However, increasing surface lipid contents and the presence of apoA-II stabilize apoA-I on the HDL particles. The reaction kinetics with lecithin:cholesterol acyltransferase are similar and slow for the isolated HDL particles, reflecting product inhibition, and/or an apoA-I conformation that is unfavorable for the activation of the lecithin:cholesterol acyltransferase reaction.

Apolipoprotein A-I↗