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PubMed1997

Combinations of the alpha-helix-turn-alpha-helix motif of TetR with respective residues from LacI or 434Cro: DNA recognition, inducer binding, and urea-dependent denaturation.

We constructed 10 different variants of TetR by substituting all or some of the residues in the alpha-helix-turn-alpha-helix (HTH) operator binding motif with the respective amino acids from LacI or 434Cro. The variants were soluble, negative transdominant over tetR in vivo, and as active as wild-type TetR in tetracycline binding in vitro. The urea-induced denaturation of the 10 variants occurs in single reversible transitions, which are centered around 4.3 M urea. Denaturation is concentration-dependent, supporting a simple two-state mechanism in which the folded dimeric protein is in equilibrium with unfolded monomers. An analysis according to the two-state model yields a Gibbs free energy of stabilization (at 0 M urea, 25 degrees C) of about 75 kJ/mol, typical for dimeric proteins of this size. Even a deletion of 24 residues from the reading head decreased the stability by only 2.7 kJ/mol. These results suggest that the DNA reading head of Tet repressor is a thermodynamically independent domain and that the thermodynamic stability of the Tet repressor dimer is determined by the association of the dimerization domains of the individual monomers. Variants containing replacements in the first alpha-helix of HTH did not show any DNA binding activity whatsoever. We attribute this to the alteration of the two N-terminal residues in this alpha-helix. TetR variants were active in nonspecific DNA binding, when either all or only the solvent-exposed residues in the recognition alpha-helix of HTH were exchanged to the respective LacI sequence. Replacement of the same residues by the respective amino acids from 434Cro yielded hybrid proteins that specifically recognize tetO in vitro. Taken together, these results establish that the similarity of operator recognition between 434Cro and TetR is greater than between TetR and LacI and confirm that prediction of the recognized DNA sequence is not obvious from the sequence of the respective HTH or recognition alpha-helix.

Amino Acid Sequence↗
PubMed2002

Novel method for selection of tRNA-driven ribozymes with enhanced stability in mammalian cells.

Intracellular stability is a critical determinant of the activity of a ribozyme in vivo. In previous studies, we succeeded in constructing an effective system for the expression of ribozymes using the promoter of a human gene for tRNA(Val). The resultant tRNA(Val)-driven ribozymes (tRNA-ribozymes) had a half-life of approximately 100 minutes. In the present study, we established a novel system for the selection of tRNA-ribozymes that were more stable than a previously generated optimally designed tRNA-ribozyme, and we confirmed that the newly selected tRNA-ribozymes worked well. Selective pressure was applied by treating cells that expressed tRNA-ribozymes with actinomycin D, and the system yielded tRNA-ribozymes with enhanced stability. The sequences isolated after selection exhibited some similarities. Furthermore, some selected tRNA-ribozymes had almost the same activity as or higher activity than that of the optimally designed tRNA-ribozyme despite the fact that the selective pressure was not aimed at enhancing the cleavage activity. Our approach might be very useful for selection not only of ribozymes with enhanced stability but also of other functional nucleic acids in vivo.

Base Sequence↗
PubMed1994

The stability and hydrophobicity of cytosolic and mitochondrial malate dehydrogenases and their relation to chaperonin-assisted folding.

mMDH and cMDH are structurally homologous enzymes which show very different responses to chaperonins during folding. The hydrophilic and stable cMDH is bound by cpn60 but released by Mg-ATP alone, while the hydrophobic and unstable mMDH requires both Mg-ATP and cpn10. Citrate equalises the stability of the native state of the two proteins but has no effect on the co-chaperonin requirement, implying that hydrophobicity, and not stability, is the determining factor. The yield and rate of folding of cMDH is unaffected while that of mMDH is markedly increased by the presence of cpn60, cpn10 and Mg-ATP. In 200 mM orthophosphate, chaperonins do not enhance the rate of folding of mMDH, but in low phosphate concentrations chaperonin-assisted folding is 3-4-times faster.

Adenosine Triphosphate↗
PubMed1981

Molecular properties of lamprey fibrinogen.

Application of conventional methods not previously used for preparing lamprey fibrinogen has yielded preparations of very high purity and stability. These preparations made possible a detailed evaluation of molecular weights in normal and chaotropic buffers. The native molecule repeatedly gave a molecular weight of 352,000 to 358,000 via sedimentation equilibrium in citrate buffer; a molecular weight of 354,500 was obtained in guanidine buffer. An anomalous increase of the value of the measured apparent partial specific volume was observed in guanidine buffer. Molecular weight data of the reduced and alkylated subunits of lamprey fibrinogen, obtained by four different methods, have led us to assign molecular weights of 110,000 for (A)alpha, 72,000 for (B) beta, and 50,000 for gamma. Based upon these molecular weights obtained for the subunits, as well as that of the native fibrinogen molecule, the subunit composition can best be fitted to the formulation [(A)n alpha, (B) beta 2, gamma 2] rather than the conventional[(A) alpha 2, (B) beta 2, gamma 2] which would yield a molecular weight of 464,000. Analysis of a stabilized clot induced by Ca2+ showed only gamma dimers; alpha subunit polymerization was undetectable. Cross-linking of lamprey fibrin in the presence of dansylcadaverine and Ca2+ results in fluorescent labeling of the gamma chains and to a lesser extent the gamma dimer. Differing from other reported vertebrate cross-linking systems, the lamprey fibrin alpha subunit appears essentially unreactive in both polymer formation and dansylcadaverine incorporation. These distinct molecular properties may be reasonably attributed to the existence of the single (A)n alpha subunit in the molecular structure of the molecule.

Amino Acids↗
PubMed2001

Optimization of production and downstream processing of the almond beta-glucosidase-mediated glucosylation of glycerol.

This article describes the synthesis of glyceryl glucoside from glycerol and glucose with almond beta-glucosidase as the catalyst. A yield of 54% (0.45 mmol/g) was obtained. The influence of the enzyme stability, the water concentration, and the water activity on the glucoside yield were determined. A molar fraction-based equilibrium constant of 2.4 +/- 0.6 was found, with which the glucoside yield could be calculated for all possible combinations of initial substrate and water fractions in the reaction mixture. A model was used to optimize the glucoside yield while minimizing one of the substrate concentrations at equilibrium. This straightforward model gives a good prediction of the measured glucoside yield, according to a parity plot.

Glucose↗
PubMed2003

Fascinating transformations of donor-acceptor complexes of group 13 metal (Al, Ga, In) derivatives with nitriles and isonitriles: from monomeric cyanides to rings and cages.

Formation of the donor-acceptor complexes of group 13 metal derivatives with nitriles and isonitriles X(3)M-D (M = Al,Ga,In; X = H,Cl,CH(3); D = RCN, RNC; R = H,CH(3)) and their subsequent reactions have been theoretically studied at the B3LYP/pVDZ level of theory. Although complexation with MX(3) stabilizes the isocyanide due to the stronger M-C donor-acceptor bond, this stabilization (20 kJ mol(-1) at most) is not sufficient to make the isocyanide form more favorable. Relationships between the dissociation enthalpy DeltaH degrees (298)(diss), charge-transfer q(CT), donor-acceptor bond energy E(DA), and the shift of the vibrational stretching mode of the CN group upon coordination Deltaomega(CN) have been examined. For a given metal center, there is a good correlation between the energy of the donor-acceptor bond and the degree of a charge transfer. Prediction of the DeltaH degrees (298)(diss) on the basis of the shift of CN stretching mode is possible within limited series of cyanide complexes (for the fixed M,R); in contrast, complexes of the isocyanides exhibit very poor Deltaomega(CN) - DeltaH degrees (298)(diss) correlation. Subsequent X ligand transfer and RX elimination reactions yielding monomeric (including donor-acceptor stabilized) and variety of oligomeric cage and ring compounds with [MN]n, [MC]n, [MNC]n cores have been considered and corresponding to thermodynamic characteristics have been obtained for the first time. Monomeric aluminum isocyanides X(2)AlNC are more stable compared to Al-C bonded isomers; for gallium and indium situation is reversed, in qualitative agreement with Pearson's HSAB concept. Substitution of X by CN in MX(3) increases the dissociation enthalpy of the MX(2)CN-NH(3) complex compared to that for MX(3)-NH(3), irrespective of the substituent X. Mechanisms of the initial reaction of the X transfer have been studied for the case X = R = H. The process of hydrogen transfer from the metal to the carbon atom in H(3)M-CNH is thermodynamically favorable and is likely to be intramolecular. By contrast, intramolecular hydrogen transfer in H(3)M-NCH has been definitely ruled out. Head-to-tail dimeric species [H(3)M-(NC)H](2) are formed exothermically and exhibit low H.H distances, which can assist in hydrogen transfer, and are likely to be the starting point for H(2) elimination. Elimination of H(2), CH(4), and C(2)H(6) from X(3)M-(NC)R adducts is very favorable thermodynamically; by contrast, elimination of HCl and CH(3)Cl is highly unfavorable even if formation of oligomer species takes place. Thus, high-temperature generation of gas-phase rings and clusters has been predicted viable in the cases X = H,CH(3) and their presence in the reactor media should not be neglected. Moderate stability of [HMCH(2)NH](4) clusters (especially in the cases M = Ga, In) makes these species viable intermediates of gas-phase reactions. Their formation may be responsible for the carbon contamination in the course of metal organic chemical vapor deposition processes of group 13 binary nitrides.

Journal Article↗
PubMed2002

Intercalators. 1. Nature of stacking interactions between intercalators (ethidium, daunomycin, ellipticine, and 4',6-diaminide-2-phenylindole) and DNA base pairs. Ab initio quantum chemical, density functional theory, and empirical potential study.

Properties of isolated intercalators (ethidium (E), daunomycin (D), ellipticine (EL), and 4,6'-diaminide-2-phenylindole (DAPI)) and their stacking interactions with adenine...thymine (AT) and guanine...cytosine (GC) nucleic acid base pairs were investigated by means of a nonempirical correlated ab initio method. All intercalators exhibit large charge delocalization, and none of them (including the DAPI dication) exhibits a site with dominant charge. All intercalators have large polarizability and are good electron acceptors, while base pairs are good electron donors. MP2/6-31G*(0.25) stabilization energies of intercalator...base pair complexes are large (E...AT, 22.4 kcal/mol; D...GC, 17.8 kcal/mol; EL...GC, 18.2 kcal/mol; DAPI...GC, 21.1 kcal/mol) and are well reproduced by modified AMBER potential (van der Waals radii of intercalator atoms are enlarged and their energy depths are increased). Standard AMBER potential underestimates binding, especially for DAPI-containing complexes. Because the DAPI dication is the best electron acceptor (among all intercalators studied), this difference is explained by the importance of the charge-transfer term, which is not included in the AMBER potential. For the neutral EL molecule, the standard AMBER force field provides correct results. The Hartree-Fock and DFT/B3LYP methods, not covering the dispersion energy, fail completely to reveal any energy minimum at the potential energy curve of the E...AT complex, and these methods thus cannot be recommended for a study of intercalation process. On the other hand, an approximate version of the DFT method, which was extended to cover London dispersion energy, yields for all complexes very good stabilization energies that are well comparable with referenced ab initio data. Besides the vertical dependence of the interaction, an energy twist dependence of the interaction energy was also investigated by a reference correlated ab initio method and empirical potentials. It is concluded that, despite the cationic (E +1, D +1, DAPI +2) or polar (EL) character of the intercalators investigated, it is the dispersion energy which predominantly contributes to the stability of intercalator...base pair complexes. Any procedure which does not cover dispersion energy is thus not suitable for studying the process of intercalation.

Adenine↗
PubMed1991

Effects of spirometric administration of tobacco smoke containing varying amounts of nicotine on human punished and non-punished operant responding.

Lever pulling of male tobacco smokers was maintained by a variable interval 20sec schedule (VI 20) of point presentation. In experiment 1 the rate of lever pulling was suppressed by a punishment contingency which stipulated that lever pulls would produce point subtractions on a variable ratio 30 schedule (VR 30). In experiment 2 the punishment contingency was omitted. Each subject participated in eight sessions each day (Mon. through Fri.). Each block of two 20-min sessions was separated by a period during which tobacco smoke containing varying amounts of nicotine was administered by the spirometry method, which ensured a constant puff volume and deep inhalation. The tobacco smoke conditions were: (1) 0.3mg nicotine yield cigarettes (baseline condition), (2) 1.2mg nicotine yield cigarettes, (3) 2.7mg nicotine yield cigarettes and (4) a condition in which room-temperature air was administered. Subjects remained at baseline conditions until responding stabilized and were then exposed to higher nicotine yield smoke or air for an entire day. Subjects were then returned to baseline conditions before administration of higher nicotine yield smoke or air. In experiment 1 (punishment contingency) lever pulling decreased as a function of increasing nicotine content in tobacco smoke. This effect upon responding was similar to the effects of CNS stimulants on punished responding in non-human subjects. In experiment 2 (no punishment contingency), no consistent effect of tobacco smoke or non-punished responding was observed.

Journal Article↗
PubMed2002

Development of an ultrahigh-temperature process for the enzymatic hydrolysis of lactose. IV. Immobilization of two thermostable beta-glycosidases and optimization of a packed-bed reactor for lactose conversion.

Recombinant hyperthermostable beta-glycosidases from the archaea Sulfolobus solfataricus (Ss beta Gly) and Pyrococcus furiosus (CelB) were covalently attached onto the insoluble carriers chitosan, controlled pore glass (CPG), and Eupergit C. For each enzyme/carrier pair, the protein-binding capacity, the immobilization yield, the pH profiles for activity and stability, the activity/temperature profile, and the kinetic constants for lactose hydrolysis at 70 degrees C were determined. Eupergit C was best among the carriers in regard to retention of native-like activity and stability of Ss beta Gly and CelB over the pH range 3.0-7.5. Its protein binding capacity of approximately 0.003 (on a mass basis) was one-third times that of CPG, while immobilization yields were typically 80% in each case. Activation energies for lactose conversion by the immobilized enzymes at pH 5.5 were in the range 50-60 kJ/mol. This is compared to values of approximately 75 kJ/mol for the free enzymes. Immobilization expands the useful pH range for CelB and Ss beta Gly by approximately 1.5 pH units toward pH 3.5 and pH 4.5, respectively. A packed-bed enzyme reactor was developed for the continuous conversion of lactose in different media, including whey and milk, and operated over extended reaction times of up to 14 days. The productivities of the Eupergit C-immobilized enzyme reactor were determined at dilution rates between 1 and 12 h(-1), and using 45 and 170 g/L initial lactose. Results of kinetic modeling for the same reactor, assuming plug flow and steady state, suggest the presence of mass-transfer limitation of the reaction rate under the conditions used. Formation of galacto-oligosaccharides in the continuous packed-bed reactor and in the batch reactor using free enzyme was closely similar in regard to yield and individual saccharide components produced.

Animals↗
PubMedDate not supplied

An in vitro biomechanical investigation of an equine interlocking nail.

OBJECTIVE: To determine the mechanical properties of Equine Interlocking Nail (EIN; JD Wheat Veterinary Orthopedic Research Laboratory, University of California, Davis) stabilized osteotomized tibiae and compare these variables with estimated in vivo loads. STUDY DESIGN: In vitro biomechanical investigation. ANIMALS: Twelve adult equine cadaveric tibiae. SAMPLE POPULATION: EIN-stabilized tibiae were tested monotonically under compression, 3- and 4-point bending, and torsion. Mechanical properties were compared with estimated in vivo loads. RESULTS: EIN-tibial composite mean compressive yield load (11 kN) and bending moment (216 Nm) were greater than loads expected postoperatively in vivo; however, the mean torsional yield load (156 Nm) was less than that expected in vivo. CONCLUSIONS: EIN-stabilized tibiae had compressive and bending strengths greater than those expected to maintain stability during walking in adult horses. Torsional yield strength did not appear sufficient to provide stability during walking in vivo. CLINICAL RELEVANCE: The EIN is not a feasible method of fracture repair for adult equine tibial fractures at this time, because its mechanical properties appear inadequate to withstand the postoperative torsional loads encountered during walking. Because this method of fracture repair may offer biological advantages, further modification of an interlocking nail for adult horses appears warranted.

Animals↗
PubMed1999

Development of sensors for direct detection of organophosphates. Part I: Immobilization, characterization and stabilization of acetylcholinesterase and organophosphate hydrolase on silica supports.

Biosensors for organophosphates in solution may be constructed by monitoring the activity of acetylcholinesterase (AChE) or organophosphate hydrolase (OPH) immobilized to a variety of microsensor platforms. The area available for enzyme immobilization is small (< 1 mm2) for microsensors. In order to construct microsensors with increased surface area for enzyme immobilization, we used a sol-gel process to create highly porous and stable silica matrices. Surface porosity of sol-gel coated surfaces was characterized using scanning electron microscopy; pore structure was found to be very similar to that of commercially available porous silica supports. Based upon this analysis, porous and non-porous silica beads were used as model substrates of sol-gel coated and uncoated sensor surfaces. Two different covalent chemistries were used to immobilize AChE and OPH to these porous and non-porous silica beads. The first chemistry used amine-silanization of silica followed by enzyme attachment using the homobifunctional linker glutaraldehyde. The second chemistry used sulfhydryl-silanization followed by enzyme attachment using the heterobifunctional linker N-gamma-maleimidobutyryloxy succinimide ester (GMBS). Surfaces were characterized in terms of total enzyme immobilized, total and specific enzyme activity, and long term stability of enzyme activity. Amine derivitization followed by glutaraldehyde linking yielded supports with greater amounts of immobilized enzyme and activity. Use of porous supports not only yielded greater amounts of immobilized enzyme and activity, but also significantly improved long term stability of enzyme activity. Enzyme was also immobilized to sol-gel coated glass slides. The mass of immobilized enzyme increased linearly with thickness of coating. However, immobilized enzyme activity saturated at a porous silica thickness of approximately 800 nm.

Acetylcholinesterase↗
PubMed2002

Factors involved in the formation of amorphous and crystalline calcium carbonate: a study of an ascidian skeleton.

The majority of invertebrate skeletal tissues are composed of the most stable crystalline polymorphs of CaCO(3), calcite, and/or aragonite. Here we describe a composite skeletal tissue from an ascidian in which amorphous and crystalline calcium carbonate coexist in well-defined domains separated by an organic sheath. Each biogenic mineral phase has a characteristic Mg content (5.9 and 1.7 mol %, respectively) and concentration of intramineral proteins (0.05 and 0.01 wt %, respectively). Macromolecular extracts from various biogenic amorphous calcium carbonate (ACC) skeletons are typically glycoproteins, rich in glutamic acid and hydroxyamino acids. The proteins from the crystalline calcitic phases are aspartate-rich. Macromolecules extracted from biogenic ACC induced the formation of stabilized ACC and/or inhibited crystallization of calcite in vitro. The yield of the synthetic ACC was 15-20%. The presence of Mg facilitated the stabilization of ACC: the protein content in synthetic ACC was 0.12 wt % in the absence of Mg and 0.07 wt % in the presence of Mg (the Mg content in the precipitate was 8.5 mol %). In contrast, the macromolecules extracted from the calcitic layer induced the formation of calcite crystals with modified morphology similar to that expressed by the original biogenic calcite. We suggest that specialized macromolecules and magnesium ions may cooperate in the stabilization of intrinsically unstable amorphous calcium carbonate and in the formation of complex ACC/calcite tissues in vivo.

Animals↗
PubMed2001

Synthesis of meso-substituted chlorins via tetrahydrobilene-a intermediates.

Chlorin building blocks incorporating a geminal dimethyl group in the reduced ring and synthetic handles in specific patterns at the perimeter of the macrocycle are expected to have utility in biomimetic and materials chemistry. A prior route employed condensation of a dihydrodipyrrin (Western half) and a bromodipyrromethane-monocarbinol (Eastern half), followed by oxidative cyclization of the putative dihydrobilene-a to form the meso-substituted zinc chlorin in yields of approximately 10%. The limited stability of the dihydrodipyrrin precluded study of the chlorin-forming process. We now have refined this methodology. A tetrahydrodipyrrin Western half (2,3,4,5-tetrahydro-1,3,3-trimethyldipyrrin) has been synthesized and found to be quite stable. The condensation of the Western half and an Eastern half (100 mM each) proceeded smoothly in CH(3)CN containing 100 mM TFA at room temperature for 30 min. The resulting linear tetrapyrrole, a 2,3,4,5-tetrahydrobilene-a, also is quite stable, enabling study of the conversion to chlorin. Refined conditions for the oxidative cyclization were found to include the following: the tetrahydrobilene-a (10 mM), AgTf (3-5 molar equiv), Zn(OAc)(2) (15 molar equiv), and 2,2,6,6-tetramethylpiperidine (15 molar equiv) in CH(3)CN at reflux exposed to air for 4-6 h, affording the zinc chlorin. The chlorin-forming process could be implemented in either a two-flask process or a one-flask process. The two-flask process was applied to form six zinc chlorins bearing substituents such as pentafluorophenyl, 3,5-di-tert-butylphenyl, TMS-ethyl benzoate, iodophenyl, or ethynylphenyl (deprotection of the TMS-ethynyl group occurred during the oxidative cyclization process). The stepwise yields (isolated) for the condensation and oxidative cyclization processes forming the tetrahydrobilene and zinc chlorin were 32-72% and 27-62%, respectively, giving overall yields of zinc chlorin from the Eastern and Western halves of 12-45%. Taken together, the refinements introduced enable 100-mg quantities of chlorin building blocks to be prepared in a facile and rational manner.

Photochemistry↗
PubMed1997

Stability of the Professional Development Assessment.

The Professional Development Assessment was constructed and pilot-tested with 76 students in three occupational therapy programs. A comparison of pretest and posttest scores yielded a significant correlation of .48, supporting the stability of responding over 1 to 2 years and suggesting usefulness of further development for evaluation of professional behaviors in students.

Adult↗
PubMed1985

Upper critical temperatures and forced ventilation effects for high-yielding dairy cows in a subtropical climate.

Upper limit of thermal stability and subsequent rise of thermoregulatory functions as affected by forced ventilation were examined. Rectal temperature, respiratory frequency, ear skin temperature, body weight, and milk yield were recorded biweekly July to March over 2 yr for 170 Israeli-Holstein cows (305-day milk yield 9000 kg/cow) at air temperatures 10 to 36 degrees C. Cows were in an open shelter. One side was force ventilated over 2.5 m along the stanchions (air velocity 1.5 to 3 m/s) from 0500 to 2200 h. Control side mean air velocity was .5 m/s. Within the 10 to 24 degrees C range, rectal temperature was not affected by air temperature or forced ventilation but increased by .02 degrees C/kg fat-corrected milk in animals producing above 24 kg/day. Between 26 and 36 degrees C rectal temperature increased with air temperature in both groups; rate of rise was halved by forced ventilation. In this range of air temperature, rectal temperature increased with rising milk yield, as in the lower air temperature range, in both high-producing and lower-producing cows in forced ventilation. Body weight or parity did not have significant effects. Mean ear skin temperature was higher for control animals, but its rate of increase with air temperature was similar in both groups. Forced ventilation reduced mean respiratory rate. An upper critical temperature is 25 to 26 degrees C and is independent of milk yield or acclimatizational state of cows exposed to the natural sequence of climate.

Animals↗
PubMed2005

[Can the cutting performance of trephines still be improved?].

PURPOSE: Today, for the preparation of donor and recipient cornea during keratoplasty either single-use or reusable metal trephines are commonly used although excimer and femtosecond laser systems are available. Due to the surgical techniques as well as the configuration of metal trephine's cutting edge irregular corneal surfaces may be induced. These will result in a postoperative astigmatism. Contemporary manufacturing processes do allow for the production of minimal cutting edge diameters (400 nm -- 1 microm), however, this is related with a reduced mechanical stability of such delicate cutting edges. It has been observed that the cutting edge of metal blades is immediately bent with the first exposure to tissue. As a result, the cutting forces are increasing and the wound configuration is negatively effected. Due to the complex geometry of trephine cutting edges, no diamond trephines are currently available. We are investigating the cutting performance of conventional trephines that were modified using nanotechnology. METHOD: Different metal trephines were characterised using a scanning electron microscope (SEM). The diameter of the cutting edge was measured. Pig cornea were studied histologically after trepanation. Selected trephines were ion-forged in a modified PVD (physical vapour deposition) reactor using highly accelerated ions which are concentrated by magnetic fields at low temperature. The consecutive processing steps were controlled by SEM analysis of the cutting edge diameters. Randomly chosen areas of treated trephines were analysed. After processing the trephines, pig cornea were cut and analysed by histological and SEM examination. Additionally, the relevant mechanical cutting parameters of untreated as well as treated trephines were measured when penetrating into a PU (polyurethane) foil. RESULTS: Preliminary microscopic analysis already reveals differences in the quality of the cutting edges. The nanotechnological modification of the cutting edges does not result in larger diameters. Compared with the initial cutting edges, the optical analysis of the modified trephine cutting edges reveals subjectively smoother surfaces. The force measurements prove a reduction of cutting forces for the treated trephines compared to the untreated ones. The SEM analysis show for the cornea treated with the modified trephines a subjectively smoother surface. Histology reveals that the surfaces of the treated cornea do not exhibit a smooth and unidirectional structure. This divergence became larger when untreated trephines were used. The mechanical characterisation of the treated trephines resulted in harder and longer-lasting surfaces. DISCUSSION: The newly developed ion-forging of trephines in combination with nanostructured carbon coatings yield complex cutting edges of higher stability with diamond-like properties. This study shows that the performance of laser trepanation cannot be reached; however, the use of nanotechnology can further improve manual trephine techniques.

Animals↗
PubMed1984

Polysaccharide storage in different streptococci.

Calf serum, pooled human plasma, polyvinylpyrollidones (PVP) and polyethylene glycols (PEG) were required as medium components by many cultures of group A.S. pyogenes for intracellular polysaccharide accumulation. These supplements produced a stabilizing effect, probably at the cell surface, and were not associated with nutrition during intracellular iodophilic polysaccharide (IPS) storage. Medium stabilization and the presence of maltose or maltose yielding polysaccharides was found to be a property unique to the group A streptococci. Cariogenic streptococci and Streptococcus agalactive did not require medium stabilization for IPS storage although addition of PVP or PEG was stimulatory. Different degrees of IPS storage in group A.S. pyogenes was found to be strain related and probably dependent on the size and degree of branching of the storage polymer.

Culture Media↗
PubMed2000

Chemical and enzymatic stability as well as transport properties of a Leu-enkephalin analogue and ester prodrugs thereof.

The Leu-enkephalin analogue (Tyr-D-Ala-Gly-Phe-Leu-NH(2)) was synthesized together with three esters prodrugs hereof. The prodrugs synthesized were the O-acetyl, O-propionyl and O-pivaloyl99%) and in good yields (60-75%). The chemical and enzymatic stability of the prodrugs has been investigated in detail. The prodrugs studied are quite chemically stable and the degradation of the prodrugs follows the pattern previously shown for similar esters (U-shaped pH-profile; maximal stability at pH 4-5). The prodrugs are degraded quantitatively in plasma to the parent peptide with half-lives in the range 2.9 min-2.6 h. Type B esterases were shown to be involved in the degradation as the half-lives increased in the presence of paraoxon. No significant stabilization was seen in 10% porcine gut homogenate. Half-lives in the same order were seen for the analogue and the prodrugs in pure Leucine aminopeptidase solution. The analogue was stable in Carboxypeptidase A solution whereas a faster degradation of the prodrugs was seen in this media. Furthermore the transport properties of the compounds has been studied. A P(app) value of 0.284x10(-6) cm/s for the analogue was obtained for the transport across Caco-2 cell monolayers in the BL-AP direction. The P(app) values were increased by a factor of 2, 7 and 18 for the acetyl-, propionyl- and pivaloylprodrug. The increase could be explained by higher lipofilicities of the prodrugs compared to the analogue.

Biological Transport↗