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At least 235 records · Page 13Linked to original sources
PubMed2000

Impact of phosphorothioate substitutions on the thermodynamic stability of an RNA GAAA tetraloop: an unexpected stabilization.

This study analyzes the impact of phosphorothioate substitutions on the thermodynamic stability of a 12-nt RNA hairpin containing a (5')GAAA(3') tetraloop. The thermodynamic consequences of stereospecific phosphorothioate substitutions 5' to each adenosine in the loop region are measured using optical melting and calorimetry experiments. Surprisingly, a single stereospecific phosphorothioate substitution 5' to the second adenosine of the tetraloop, R(p)-A7, results in a stabilization corresponding to a Delta(DeltaG(37)(degrees)(C)) of approximately -2.9 kcal mol(-1) (0.1 M NaCl) when compared with that of an unmodified sample. Five other phosphorothioate-substituted samples did not show significant thermodynamic differences in comparison with the unsubstituted samples. Addition of Mg(2+) to all of the hairpins studied results in increased t(m's) that are fit with a general electrostatic model to a dissociation constant of K(d)(Mg(2+)) approximately 2-3 mM (0.1 M NaCl). The R(p)-A7 phosphorothioate-substituted hairpin showed an unusual decrease in t(m) and apparent increase in enthalpy of unfolding upon addition of Cd(2+). These results may impact the interpretation of interference mapping experiments that use phosphorothioate substitutions to characterize RNAs in solution.

Cadmium↗
PubMed1994

A model for the stabilities of RNA hairpins based on a study of the sequence dependence of stability for hairpins of six nucleotides.

Thermodynamic parameters are reported for hairpin formation in 1 M NaCl by RNA sequences of the type GGCXUAAUYGCC, where XY is the set of 10 possible mismatch base pairs. A nearest neighbor analysis of the data indicates the free energy for loop formation at 37 C varies from 2.9 to 4.5 kcal/mol. Thermodynamic parameters are also reported for hairpin formation by RNA sequences of the type GGXGUAAUAYCC (where XY are CG, GC, AU, UA, GU, and UG), with the common naturally occurring GA first mismatch (45% of small and large subunit rRNA loops of six). These results allow the development of a model to predict the stability of RNA hairpin loops. The model includes the size of the loop, the identity of the closing base pair, the free energy increment (delta G zero 37MM) for interaction of the closing base pair with the first mismatch, and an additional stabilization term for GA and UU first mismatches. delta G zero 37L(n) = delta G zero 37i(n) + delta G zero 37MM + 0.4 (if closed by AU or UA) -0.7 (if first mismatch is GA or UU). Here delta G zero 37i(n) is the free energy for initiating a loop of n nucleotides. delta G zero 37i(n) for n = 4-9 is 4.9, 4.4, 5.0, 5.0, 5.1, and 5.2 kcal/mol, respectively. The delta G zero 37MM is derived from measurements of model duplexes with terminal mismatches. The model gives good agreement when tested against four naturally occurring hairpin sequences.

Base Sequence↗
PubMed1998

Sequence requirements for stabilization of a peptide reverse turn in water solution--proline is not essential for stability.

To probe the sequence requirements for stabilization of a reverse turn conformation in a short peptide in water solution, the behavior of two series of peptides was investigated by nuclear magnetic resonance (NMR) spectroscopy. The peptides have the general sequences XPGDV and AXGDV, where X is a representative subset of all 20 naturally occurring amino acids. The residues chosen at positions 3 and 4, Gly and Asp, respectively, were shown to give the greatest population of reverse turns in a previous study [Dyson, H. J., Rance, M., Houghten, R. A., Lerner, R. A. & Wright, P. E. (1988) J. Mol. Biol. 201, 161-200]. Within this framework, the identity of the first residue of the turn (X in XPGDV) does not greatly influence the turn population, although a small but significant increase is observed for residues such as Ala which have a preference for backbone conformations in the alpha region of (phi,psi) space. The series AXGDV was initially studied for completeness only, since it was expected that the turn would not be stabilized in such a small linear peptide in the absence of proline. In contrast, it appears that a significant population of type II turn conformations is to be found in peptides in the series AXGDV, although proline remains one of the most favorable residues at position 2. These results indicate that while residues at all positions within the turn can influence the turn population, the presence of Gly-Asp as the third and fourth members of the sequence gives a strong bias towards type II turn formation regardless of the residues at positions 1 and 2. Our results give a final prediction that the sequence with the highest intrinsic propensity for turn formation is APGD.

Amino Acid Sequence↗
PubMed2006

Factors affecting stability of z-ligustilide in the volatile oil of radix angelicae sinensis and ligusticum chuanxiong and its stability prediction.

The purpose of this investigation is to obtain a suitable vehicle for Z-ligustilide in the volatile oil of Radix Angelicae Sinensis and Ligusticum Chuanxiong in which it is stable enough for the application in pharmaceutics, to investigate its degradation laws, and to predict its shelf-life at 25 degrees C. Factors including temperature, light, pH value, co-solvents and antioxidants can all influence the stability of Z-ligustilide, thereinto antioxidants could markedly improve its stability in aqueous solution by almost 35%. The suitable vehicle for Z-ligustilide contains 1.5% tween-80, 0.3% Vitamin C, and 20% propylene glycol (PG). Furthermore, the degradation rates of Z-ligustilide were found to conform to a rate equation following Weibull probability distribution within a range of degradation ratio, and the equation could be expressed as follow: ln ln (1/1-alpha) = ln k + m ln t. Where alpha is degradation ratio; t is time; m and k are constants relating to the degradation rate. The degradation rate will get greater as the increasing of parameter k. According to the degradation law obtained from the equation, the drug shelf-life (10% of active ingredient degraded, T90) in this vehicle was predicted to be more than 1.77 years at 25 degrees C through Arrehenius equation and accelerating experiments. The present investigation was undertaken to propose a kinetic treatment that may be applicable to any type of degradation of the active ingredient of pharmaceutical formulation, and also could provide a good foundation for the new drug development of Z-ligustilide, especially for injection formulation.

4-Butyrolactone↗
PubMed2003

Polymer-stabilized emulsion systems: structural characteristics and physical stability evaluation.

In the course of investigation of the effects of polymeric emulsifier (Simulgel A) on the structural characteristics and physical stability (during storage time) of semisolid o/w emulsion systems with different emulsifier concentrations, dispersed droplets of an internal oil phase (4.5-11.5 microm size) were discovered. The degree of dispersity was increased along with increasing polymeric emulsifier concentration (from 1 to 5% m/m), which was shown by microscopic analysis. Experiments were also carried out using pH, electrical conductivity, and rheological (rotational and oscillatory) measurements. Emulsion samples expressed a mild acid value acceptable for topical preparations during a 6-month period. Electrical conductivity of fresh samples ranged from 0.423 to 2.009 mS/cm and after a 6-month period this slightly increased to 0.663-2.890 mS/cm. All emulsion samples revealed plastic flow behavior in fresh state, with changes in flow pattern during storage time. Viscoelastic properties defined by the oscillatory rheometry results were in good correlation with parameters of rotational rheometry and degree of internal phase dispersity. High yield stress values (flow tests) and favorable low values of damping factor (oscillatory studies) both indicated good internal structuring and long-term stability of sample containing 3% m/m of polymeric emulsifier.

Acrylic Resins↗
PubMed1995

Relative stabilities of DNA three-way, four-way and five-way junctions (multi-helix junction loops): unpaired nucleotides can be stabilizing or destabilizing.

Competition binding and UV melting studies of a DNA model system consisting of three, four or five mutually complementary oligonucleotides demonstrate that unpaired bases at the branch point stabilize three- and five-way junction loops but destabilize four-way junctions. The inclusion of unpaired nucleotides permits the assembly of five-way DNA junction complexes (5WJ) having as few as seven basepairs per arm from five mutually complementary oligonucleotides. Previous work showed that 5WJ, having eight basepairs per arm but lacking unpaired bases, could not be assembled [Wang, Y.L., Mueller, J.E., Kemper, B. and Seeman, N.C. (1991) Biochemistry, 30, 5667-5674]. Competition binding experiments demonstrate that four-way junctions (4WJ) are more stable than three-way junctions (3WJ), when no unpaired bases are included at the branch point, but less stable when unpaired bases are present at the junction. 5WJ complexes are in all cases less stable than 4WJ or 3WJ complexes. UV melting curves confirm the relative stabilities of these junctions. These results provide qualitative guidelines for improving the way in which multi-helix junction loops are handled in secondary structure prediction programs, especially for single-stranded nucleic acids having primary sequences that can form alternative structures comprising different types of junctions.

Autoradiography↗
PubMed1990

Correlation between stability of a protein and its dipeptide composition: a novel approach for predicting in vivo stability of a protein from its primary sequence.

Statistical analysis of 12 unstable and 32 stable proteins revealed that there are certain dipeptides, the occurrence of which is significantly different in the unstable proteins compared with those in the stable ones. Based on the impact of these dipeptides on the unstable proteins over the stable ones, a weight value of instability is assigned to each of the dipeptides. For a given protein the summation of these weight values normalized to the length of its sequence helps to distinguish between unstable and stable proteins. Results suggest that the in vivo instability of proteins is possibly determined by the order of certain amino acids in its sequence. An attempt is made to correlate metabolic stability of proteins with features of their primary sequence where weight values of instability for a protein of known sequence could thus be used as an index for predicting its stability characteristics.

Amino Acid Sequence↗
PubMed1995

The effect of lysine, a water-structure breaker, on the stability of phospholipid-stabilized emulsions.

Phospholipid-stabilized emulsion properties were studied in the presence of lysine, a water-structure breaker, using two unrelated procedures, photon correlation spectroscopy and a light obscuration instrument. Commercial Intralipid was used as a control. Lysine 0.125, 0.25 and 0.5 M induced changes in the size distribution of a non-heated model emulsion system, irrespective of any changes produced by environmental pH. Some of the laboratory-prepared emulsions containing lysine were more stable than the corresponding commercial heat-sterilized product Intralipid, once heated. The results suggest that lysine is producing an effect on the nascent oil-water interface that controls the physical stability of the system. Once the heat-induced interfacial rearrangement of the individual phospholipid molecules occurs, the influence of lysine becomes diminished.

Centrifugation↗
PubMed2006

Mitogen-activated protein kinase-activated protein kinase 2 regulates tumor necrosis factor mRNA stability and translation mainly by altering tristetraprolin expression, stability, and binding to adenine/uridine-rich element.

The mitogen-activated protein kinase (MAPK) p38/MAPK-activated protein kinase 2 (MK2) signaling pathway plays an important role in the posttranscriptional regulation of tumor necrosis factor (TNF), which is dependent on the adenine/uridine-rich element (ARE) in the 3' untranslated region of TNF mRNA. After lipopolysaccharide (LPS) stimulation, MK2-deficient macrophages show a 90% reduction in TNF production compared to the wild type. Tristetraprolin (TTP), a protein induced by LPS, binds ARE and destabilizes TNF mRNA. Accordingly, macrophages lacking TTP produce large amounts of TNF. Here, we generated MK2/TTP double knockout mice and show that, after LPS stimulation, bone marrow-derived macrophages produce TNF mRNA and protein levels comparable to those of TTP knockout cells, indicating that in the regulation of TNF biosynthesis TTP is genetically downstream of MK2. In addition, we show that MK2 is essential for the stabilization of TTP mRNA, and phosphorylation by MK2 leads to increased TTP protein stability but reduced ARE affinity. These data suggest that MK2 inhibits the mRNA destabilizing activity of TTP and, in parallel, codegradation of TTP together, with the target mRNA resulting in increased cellular levels of TTP.

3' Untranslated Regions↗
PubMedDate not supplied

Effects of diethylenetriamine and Aerosol OT on the stability of oil-in-water emulsion stabilized by interfacial polyurea film.

The stability of oil-in-water emulsions containing a triisocyanate soluble in the oil phase and an amine in the water phase was investigated. The oil component was di-n-butyl phthalate (DBP) containing Aerosol OT as an emulsifier. The time required for the average size parameter to reach a constant value was studied. It was found that the polyurea film produced by an interfacial polymerization reaction between the amine and the triisocyanate contributed to forming a stable emulsion at a lower Aerosol OT concentration, but at a higher Aerosol OT concentration the amine did not show any effect on the emulsion stability.

Chemical Phenomena↗
PubMed2003

International Conference on Harmonisation; Stability Data Package for Registration Applications in Climatic Zones III and IV; Stability Testing of New Drug Substances and Products; availability. Notice.

The Food and Drug Administration (FDA) is announcing the availability of two guidances prepared under the auspices of the International Conference on Harmonization of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH). The first is a guidance entitled "Q1F Stability Data Package for Registration Applications in Climatic Zones III and IV'' (the Q1F guidance). The second is a revised guidance entitled "Q1A(R2) Stability Testing of New Drug Substances and Products'' (the Q1A guidance). The Q1F guidance, which is an annex to the Q1A guidance, defines an approach for broader use of the Q1A guidance for territories in climatic zones III and IV. The revised Q1A guidance incorporates relevant Q1F recommendations.

Climate↗
PubMed2004

Applicational possibilities of linear and non-linear (polynomial) regression and analysis of variance. III. Stability determination of pharmaceutical preparations: stability of diclofenac-sodium in Diclofen injections.

This paper presents the application of the regression analysis program and the program for comparing linear regressions (modified method for one-way, analysis of variance), writtens in BASIC program language, for instance, determination of content of Diclofenac-Sodium (active ingredient in DIKLOFEN injections, ampules á 75 mg/3 ml). Stability testing of Diclofenac-Sodium was done by isothermic method of accelerated aging at 4 different temperatures (30 degrees, 40 degrees, 50 degrees and 60 degrees C) as a function of time (4 different duration of treatment: (0-155, 0-145, 0-74 and 0-44 days). The decrease in stability (decrease in the mean value of the content of Diclofenac-Sodium (in %), at different temperatures as a function of time, is possible to describe by, linear dependance. According to the value for regression equation values, the times are assessed in which the content of Diclofenac-Sodium (in %) will decrease by 10%, of the initial value. The times are follows at 30 degrees C 761.02 days, at 40 degrees C 397.26 days, at 50 degrees C 201.96 days and at 60 degrees C 58.85 days. The estimated times (in days) in which the mean value for Diclofenac-Sodium content (in %) will by 10% of the initial values, as a junction of time, are most suitably described by 3rd order parabola. Based on the parameter values which describe the 3rd order parabola, the time was estimated in which Diclofenac-Sodium content mean value (in %) will fall by 10% of the initial one at average ambient temperatures of 20 degrees C and 25 degrees C. The times are: 1409.47 days (20 degrees C) and 1042.39 days (25 degrees C). Based on the value for Fischer's coefficien (F), the comparison of trenf of Diclofenac-Sodium content (in %) shows that, under the influence of different temperatures as a function of time, among them, depending on temperature value, there is: statistically very significant difference (P << .05) at 50 degrees C and lower toward 60 degrees C, i.e. statistically probably significant difference (P > 0.01) at 40 degrees C and lower towards 50 degrees C and there is no statistically significance difference (P >> 0.05) at 30 degrees C towards 40 degrees C.

Algorithms↗
PubMedDate not supplied

Thermal stability of penicillin G acylase : effect of stabilizing agents.

The role of sugars, polyhydroxy compounds, phenylacetic acid and 6-aminopenicillanic acid in stabilization of immobilized penicillin G acylase (IMPGA) was studied. The loss in the activity of IMPGA at 50 degrees C, 2 h, after incorporation of sucrose and mannitol at 0.1 M concentration was 16 and 18% respectively; the loss in the activity of the enzyme under these conditions in the absence of stabilizing agents was 40%.

Carbohydrates↗
PubMed1989

[Effect of chemical structure of various penicillins on the stability of beta-lactam group in their molecules. VI. Effects of relative humidity and heat on the stability of amoxicillin trihydrate and amoxicillin sodium in the solid phase].

Dependence of stability of Amoxycillin trihydrate (AMOXY. 3H2O) and Amoxycillin-Natrium (AMOXY-Na) upon relative humidity (RH) and temperature was investigated using the iodometric method for the determination of Amoxycillin. The first order rate constants and activation parameters have been determined at controlled moisture conditions. The straightline relationship lg k = f(RH) at T = const. has been stated, which however is fulfilled by AMOXY-Na only at RH greater than or equal to 50%. The calculated factors of the equation lg k = a + b.1/T + c.RH proved to enable estimation of the stability of the studied antibiotics at any climate conditions. The performed kinetic studies have shown that AMOXY-Na undergoes degradation about 1.5 x 10(3) times faster than AMOXY.3H2O, at RH greater than or equal to 50%.

Amoxicillin↗
PubMed1986

[The stability of hydrochlorothiazide and cyclopenthiazide in various dosage forms. 3. Stability of hydrochlorothiazide injection solutions].

Studies of the stability of hydrochlorothiazide (1) in N-methylacetamide prove that a temperature dependent equilibrium is found in the isothermic short-term test as well as in the longterm stability test. Whereas the kinetic reaction evaluation after Arrhenius doesn't allow a forecast, the 25 degrees C-values, extrapolated after the Van't Hoff-equation, were confirmed using the long-term test. By adding the decomposition product aminodisulfamide (2) the hydrolysis is restricted.

Cyclopenthiazide↗
PubMed1983

[Analysis and stability of suxamethonium chloride. 2: Study of various factors in the stability of suxamethonium chloride injection solutions].

The packing material (glass ampoules and plastic disposable syringes) and the degree of purity of the active agent do not affect the stability of injectable suxamethonium chloride solutions. In contrast to this, the stability was markedly improved by lowering the temperature of storage. Injectable suxamethonium chloride solutions may be stored in plastic disposable syringes.

Chemistry, Pharmaceutical↗
PubMed1983

[Analysis and stability of atropine sulfate and scopolamine hydrobromide. 3: Studies on the stability of tropane alkaloids; 80: contribution to problems in the use of plastic containers for liquid pharmaceuticals].

On evaluating their investigations, the authors discuss the degradation mechanism of tropane alkaloids in aqueous solution. As evidenced by competitive and consecutive reactions (hydrolysis, dehydration, dimerization), the degradation mechanism is characterized by a complicated course. Apart from temperature, a series of other factors, above all the pH value, exerts a considerable effect on the stability of both the alkaloids. Unbuffered solutions are more stable than buffered ones. The isothermal short-time test is not suitable for stability prediction. In the long-time test it was found that unbuffered aqueous atropine sulphate solutions still contained 90% of their initial drug content after 18 months' storage in low-density polyethylene containers at 20 degrees C. Scopolamine hydrobromide solutions showed an almost analogous behaviour. Of the container materials tested, low-density polyethylene bottles have been found to be best suited.

Atropine↗
PubMed1995

In vivo stability and disposition of a self-stabilized oligodeoxynucleotide phosphorothioate in rats.

The use of antisense oligonucleotides represents a novel, genetically based therapy. The biostability and pharmacokinetics of a 33-mer self-stabilized oligodeoxynucleotide with significant anti-HIV activity was determined in rats after intravenous administration of [35S]oligodeoxynucleotide. Plasma disappearance of the labeled oligodeoxynucleotide could be described by a two-compartment model, with half-lives of 0.54 and 41.44 h. The oligodeoxynucleotide in plasma remained mainly intact. Urinary excretion represented the major elimination pathway, with approximately 27% of the administered dose excreted within 24 h and 57% over 240 h. The majority of radioactivity in urine was attached to degradative products. Fecal excretion was a minor elimination pathway. A wide tissue distribution of the oligonucleotide was observed, with the majority of radioactivity in most tissues being intact. Compared with other linear oligonucleotide phosphorothioates, the self-stabilized oligonucleotide was more stable in vivo, which may be important in development of antisense oligonucleotides as therapeutic agents.

Animals↗