Comparative Evaluation of Ion Release in Bonded and Nonbonded Stainless Steel Brackets with Use of Different Mouthwashes: An In vitro Study.

BACKGROUND: Orthodontic treatment is a long-term procedure that involves the usage of brackets and archwires which are mainly metallic in nature. This study evaluated the levels of metal ions release from bonded and nonbonded orthodontic brackets after immersion in distilled deionized water and three different types of mouthwash. MATERIALS AND METHODS: Eighty premolar stainless steel brackets (3M, Unitek) were divided into Group A (bonded brackets) and Group B (nonbonded brackets). Each group was further subdivided into four subgroups to analyze the release of ions from three different types of mouthwash along with the control group. All the samples were incubated at 37°C for 45 days, and immersion solutions were tested in inductively coupled plasma-optical emission spectrometer for the release of free metal ions. RESULTS: Mean ion release in the bonded bracket group was less than that of nonbonded bracket group. Ion release in control subgroup of both groups was 0.18 ± 0.08 µg/dl (A1) and 0.17 ± 0.06 µg/dl (B1); in Phos-Flur mouthwash subgroup was 0.12 ± 0.06 µg/dl (A2) and 0.13 ± 0.05 µg/dl (B2); in chlohex mouthwash subgroup was 0.13 ± 0.06 µg/dl (A3) and 0.14 ± 0.06 µg/dl (B3); in Hiora mouthwash subgroup was 0.10 ± 0.06 µg/dl (A4) and 0.12 ± 0.05 µg/dl (B4). CONCLUSIONS: The mean ion release was the highest in deionized water (control group) followed by chlohex, Phos-Flur, and Hiora in both Group A and Group B. Ion leaching from bonded brackets was less compared to nonbonded brackets in all different mouthwashes except in the control group. However, this difference was statistically insignificant (P > 0.05) between all the groups.

An esthetic treatment outcome of orthognathic surgery and dentofacial orthopedics in class II treatment: a cephalometric study.

AIM: The main objective of any orthodontic treatment is to achieve well-established stable occlusal relationship with a definite positive change in facial profile. The purpose of this study was to determine, if such a goal is achievable for patients who could be classified as borderline surgical cases without the invasive use of the actual surgery or, with the use of the recently developed and rapidly spreading fixed functional appliance system (Forsus) and a comparison of the esthetic treatment outcome with the two systems. MATERIALS AND METHODS: Twelve postadolescent borderline skeletal class II patients with a deficient mandible. All the patients used in the study were treated by a preadjusted edgewise appliance for presurgical decompensation with or without extractions and for postsurgical finishing and detailing. Out of the 12 patients six were treated with bilateral saggital split osteotomy (BSSO) and six were treated with fixed functional appliance (Forsus). RESULTS: The results suggested that although surgical patients had a better mandibular advancement, profile reduction, and marked improvements in soft tissue structures, the patients who had undergone fixed functional therapy also had comparable improvement in the above aspects. In the maxilla there was no change in cases treated with surgery but in case of Forsus some retraction of anterior dental segment was evident. CONCLUSION: In surgical group, class II malocclusion correction was more skeletal than dental, whereas in functional group class II malocclusion correction was more dental than skeletal. CLINICAL SIGNIFICANCE: Looking at the common surgical risks, cost-effective and postsurgical problems and patients with borderline class II malocclusion, fixed functional therapy is a valuable adjunct in the management of class II malocclusion.

Oxyanion-mediated inhibition of serine proteases.

Novel aryl derivatives of benzamidine were synthesized and tested for their inhibitory potency against bovine trypsin, rat skin tryptase, human recombinant granzyme A, human thrombin, and human plasma kallikrein. All compounds show competitive inhibition against these proteases with Ki values in the micromolar range. X-ray structures were determined to 1.8 A resolution for trypsin complexed with two of the para-substituted benzamidine derivatives, 1-(4-amidinophenyl)-3-(4-chlorophenyl)urea (ACPU) and 1-(4-amidinophenyl)-3-(4-phenoxyphenyl)urea (APPU). Although the inhibitors do not engage in direct and specific interactions outside the S1 pocket, they do form intimate indirect contacts with the active site of trypsin. The inhibitors are linked to the enzyme by a sulfate ion that forms an intricate network of three-centered hydrogen bonds. Comparison of these structures with other serine protease structures with noncovalently bound oxyanions reveals a pair of highly conserved oxyanion-binding sites in the active site. The positions of noncovalently bound oxyanions, such as the oxygen atoms of sulfate, are distinct from the positions of covalent oxyanions of tetrahedral intermediates. Noncovalent oxyanion positions are outside the "oxyanion hole." Kinetics data suggest that protonation stabilizes the ternary inhibitor/oxyanion/protease complex. In sum, both cations and anions can mediate Ki. Cation mediation of potency of competitive inhibitors of serine proteases was previously reported by Stroud and co-workers [Katz, B. A., Clark, J. M., Finer-Moore, J. S., Jenkins, T. E., Johnson, C. R., Ross, M. J., Luong, C., Moore, W. R., and Stroud, R. M. (1998) Nature 391, 608-612].

Novel peptidyl alpha-keto amide inhibitors of calpains and other cysteine proteases.

A series of new dipeptidyl alpha-keto amides of the general structure R1-L-Leu-D,L-AA-CONH-R2 were synthesized and evaluated as inhibitors for the cysteine proteases calpain I, calpain II, and cathepsin B. They combine 10 different N-protecting groups (R1), 3 amino acids residues in P1 (AA), and 44 distinct substituents on the alpha-keto amide nitrogen (R2). In general, calpain II was more sensitive to these inhibitors than calpain I, with a large number of inhibitors displaying dissociation constants (Ki) in the 10-100 nM range. Calpain I was also effectively inhibited, but very low Ki values were observed with a smaller number of inhibitors than with calpain II. Cathepsin B was weakly inhibited by most compounds in this study. The best inhibitors for calpain II were Z-Leu-Abu-CONH-CH2-CHOH-C6H5 (Ki = 15 nM), Z-Leu-Abu-CONH-CH2-2-pyridyl (Ki = 17 nM), and Z-Leu-Abu-CONH-CH2-C6H3(3,5(OMe)2) (Ki = 22 nM). The best calpain I inhibitor in this study was Z-Leu-Nva-CONH-CH2-2-pyridyl (Ki = 19 nM). The peptide alpha-keto amide Z-Leu-Abu-CONH-(CH2)2-3-indolyl was the best inhibitor for cathepsin B (Ki = 31 nM). Some compounds acted as specific calpain inhibitors, with comparable activity on both calpains I and II and a lack of activity on cathepsin B (e.g., 40, 42, 48, 70). Others were specific inhibitors for calpain I (e.g., 73) or calpain II (e.g., 18, 19, 33, 35, 56). Such inhibitors may be useful in elucidating the physiological and pathological events involving these proteases and may become possible therapeutic agents.

Mammalian tissue trypsin-like enzymes: substrate specificity and inhibitory potency of substituted isocoumarin mechanism-based inhibitors, benzamidine derivatives, and arginine fluoroalkyl ketone transition-state inhibitors.

Amino acid and peptide thioesters which contained Arg or Lys in the P1 position were tested as substrates for rat skin tryptase, and the kinetic constants Kcat/KM for the better substrates such as Z-Aba-Arg-SBzl, and Z-Gly-Arg-SBzl were over 5,000,000 M-1 s-1. The inhibitory potency of arginine fluoroalkyl ketones, benzamidine derivatives, and substituted isocoumarins containing basic functional groups was studied with rat skin tryptase, human lung tryptase, human skin tryptase, and bovine trypsin. 1-Naphthoyl-Arg-CF3 was the best arginine fluoroalkyl ketone reversible inhibitor for rat skin tryptase with a KI of 0.9 microM. 1-(4-Amidino-phenyl)-3-(4-phenoxyphenyl) urea showed competitive inhibition against bovine trypsin and rat skin tryptase with KI values of 2 and 4 microM, respectively. Isocoumarin derivatives with isothioureidoalkoxy substituents at the 3 position were potent irreversible inhibitors of these three tryptases with Kobs/[I] values of 10(4)-10(5) M-1 s-1. 4-Chloro-3-(2-isothioureido)ethoxy-7-phenylcarbamoylaminoisocou marin and 7-benzylcarbamoylamino-4-chloro-3-(3-isothioureido)propox yisocoumarin inactivated trypsin and formed stable trypsin-inhibitor complexes which regained less than 8% of activity upon standing in the pH 7.5 buffer and regained 30-75% of activity in the presence of 0.3 M NH2OH after 1 day. In contrast, the complexes with rat skin tryptase regained activity rapidly, indicating differences in the inhibition mechanism and active site structures of these related enzymes.

Peptide alpha-keto ester, alpha-keto amide, and alpha-keto acid inhibitors of calpains and other cysteine proteases.

A series of dipeptidyl and tripeptidyl alpha-keto esters, alpha-keto amides, and alpha-keto acids having leucine in the P2 position were synthesized and evaluated as inhibitors for the cysteine proteases calpain I, calpain II, cathepsin B, and papain. In general, peptidyl alpha-keto acids were more inhibitory toward calpain I and II than alpha-keto amides, which in turn were more effective than alpha-keto esters. In the series Z-Leu-AA-COOEt, the inhibitory potency decreased in the order: Met (lowest KI) > Nva > Phe > 4-Cl-Phe > Abu > Nle (highest KI) with calpain I, while almost the reverse order was observed for calpain II. Extending the dipeptide alpha-keto ester to a tripeptide alpha-keto ester yielded significant enhancement in the inhibitory potency toward cathepsin B, but smaller changes toward the calpains. Changing the ester group in the alpha-keto esters did not substantially decrease KI values for calpain I and calpain II. N-Monosubstituted alpha-keto amides were better inhibitors than the corresponding alpha-keto esters. alpha-Keto amides with hydrophobic alkyl groups or alkyl groups with an attached phenyl group had the lower KI values. N,N-Disubstituted alpha-keto amides were much less potent inhibitors than the corresponding N-monosubstituted peptide alpha-keto amides. The peptide alpha-keto acid Z-Leu-Phe-COOH was the best inhibitor for calpain I (KI = 0.0085 microM) and calpain II (KI = 0.0057 microM) discovered in this study. It is likely that the inhibitors are transition-state analogs and form tetrahedral adducts with the active site cysteine of cysteine proteases and form hydrogen bonds with the active site histidine and possibly another hydrogen bond donor in the case of monosubstituted amides. Several inhibitors prevented spectrin degradation in a platelet membrane permeability assay and may be useful for the treatment of diseases which involve neurodegeneration.

Surface-active substances of the guinea pig tubotympanum: a chemical and physical analysis.

An attempt to describe the nature of the surface-active substances of the eustachian tube lining layer that influence normal tubal function was undertaken. Under sterile conditions, guinea pig tubotympanic washings were collected, centrifuged, and pooled. Analysis of the pooled lavages using standard surface chemistry techniques confirmed the presence of significant surface-tension-lowering activity in the mucous lining layer of the eustachian tube, but the surface pressure obtained is neither as great nor displays the same degree of hysteresis as pulmonary surfactant. Following separation into aqueous and lipid fractions, measurable amounts of surface activity can be found in both isolates. The chemical composition and concentration of the lipid fraction, and its relative contribution to the surface activity of the tubotympanic washings, however, is smaller and radically different from the phospholipids found in surfactant. A significantly higher concentration of protein was recovered in comparison with the lipid portion, and it was observed that the surface activity of the total washings and the aqueous phase bore remarkable similarities. Although the surface-tension-lowering properties of the tubal lining layer may be the consequence of a combined synergistic action of the lipid and protein moieties, we believe that the current evidence points toward the proteins as being the primary tubal surface-tension-lowering substances.

Correlations between surface area and the rate of enzymatic desaturation with methyl branched 8, 11, 14-eicosatrienoic acid.

Methyl-branched derivatives of methyl 8,11,14-eicosatrienoate form stable liquid-expanded monolayers. Surface areas are expanded by the methyl branch. The expansion effect is a function of surface pressure. At high surface pressure, the greatest expansion occurs with a mid-point methyl branch. At low surface pressure, surface area increases continuously as the methyl group is moved along the carbon chain from carbon 19 to carbon 5. Desaturase activity varies inversely with surface area, and a linear correlation exists between surface area at low surface pressure and the desaturation rate. These data support the concept that lipid structure and its effect on short range forces between molecules is an important factor in desaturase activity.

Effects of ionization and counterion binding on the surface areas of phosphatidic acids in monolayers.

At 24-26 degrees C, force-area isotherms show that unionized dipalmitoyl phosphatidic acid forms a solid-condensed film while unionized egg and dioleoyl phosphatidic acids form liquid-expanded films. Surface area is a characteristic feature of a specific phosphatidic acid and the purity of a phosphatidic acid preparation can be established by the surface area of the unionized phosphatidic acid (acid subphase) at 17 dynes/cm (castor oil piston). Ionized dipalmitoyl phosphatidic acid desorbs from a monolayer at a measurable rate while ionized egg and dioleoyl phosphatidic acids desorb too slowly for rate studies. The apparent surface pK(2) for dipalmitoyl phosphatidic acid, calculated from desorption rates, is 9.4. Surface areas of the phosphatidic acids expand with ionization. Solid dipalmitoyl phosphatidic acid films expand only in the pK(2) region, showing one inflection point which indicates that the K(1)/K(2) ratio is less than 100 and that, as a consequence of this ratio, the apparent surface pK(1) is greater than 7.4. Liquid egg and dioleoyl phosphatidic acid films have two inflection points, expanding in both the pK(1) and pK(2) regions. The apparent surface pK(1) and pK(2) values, calculated from inflection points in surface area data, are 3.5 and 8.0, respectively. Film expansion with phosphatidate anions is less than anticipated, showing the presence of weak transient hydrogen bonds. Expanded phosphatidate anion films are condensed by alkaline earth cations. The Ca(2+) and Ba(2+) salts of completely ionized phosphatidic acids collapse from monolayers, showing that the phosphatidate anion may function as an ionophore for the transport of alkaline earth ions.-Patil, G. S., N. J. Dorman, and D. G. Cornwell. Effects of ionization and counterion binding on the surface areas of phosphatidic acids in monolayers.

Interfacial oxidation of alpha-tocopherol and the surface properties of its oxidation products.

dl-alpha-Tocopherol spread on an acidic subphase as a gaseous monolayer was oxidized slowly to a derivative that was identified by thin-layer chromatography as alpha-tocopherylquinone. The derivative generated the same II-A isotherm as alpha-tocopherylquinone. When the subphase contained gold chloride, alpha-tocopherol was oxidized rapidly and quantitatively to alpha-tocopherylquinone. dl-alpha-Tocopherol spread on a basic subphase as a gaseous monolayer was oxidized slowly to a mixture that contained alpha-tocopherol, a quinone, and a nonpolar derivative. The mixture generated a II-A isotherm with an inflection point below the equilibrium spreading pressure of either alpha-tocopherol or alpha-tocopherylquinone. When potassium ferricyanide was added to the alkaline subphase, alpha-tocopherol was oxidized rapidly to a mixture that contained both the nonpolar derivative (major product) and the quinone (minor product). The nonpolar derivative was isolated by thin-layer chromatography and identified as the spirodienone ether by ultraviolet, infrared, and chemical ionization mass spectra. The spirodienone ether had a low equilibrium spreading pressure that explained the inflection point in the II-A isotherm generated by alpha-tocopherol on an alkaline subphase. Surface area data showed that dl-alpha-tocopherol formed immiscible films with stearyl alcohol and miscible films with oleyl alcohol. II-A isotherms showed that alpha-tocopherol in both immiscible and miscible mixtures was oxidized rapidly on an alkaline potassium ferricyanide subphase to the spirodienone ether. Collapse pressure data showed that the spirodienone ether formed an immiscible film with stearyl alcohol and a miscible film with oleyl alcohol. Interfacial oxidation experiments showed that alpha-tocopherol is oxidized either to tocopherylquinone (acidic subphase) or to the spirodienone ether (alkaline subphase). The natural occurrence of both tocopherylquinone and the spirodienone ether suggests that several types of oxidant stress are found in biological systems. One type of oxidant stress may involve the peroxy radical generating tocopherylquinone; a second type may involve hydroxyl radical-hydroxide ion generating the spirodienone ether.

Surface phase separation and collapse of the stearate anion--alkaline earth cation complex.

The surface properties of fatty acid and fatty acid-alcohol mixtures were examined at 22-24 degrees C. At pH 12, sodium stearate forms a rigid surface film that generates an equilibrium spreading pressure of 16.5 dynes/cm. At pH 12, stearate-alkaline earth cation films collapse at the air-water interface and do not generate significant equilibrium spreading pressures. The rate of film collapse depends on the counterion decreasing in the sequence Ba2+ greater than Sr2+ greater than Ca2+. Stearate-stearyl alcohol mixtures form solid (condensed) films that are relatively stable and behave initially as homogeneous surfaces in their selectivities for counterions. Stearate-oleyl alcohol mixtures form fluid (expanded) films that are unstable. Lateral phase separations occur rapidly in fluid films and the stearate-alkaline earth cation phase collapses. The rate of film collapse in the fluid mixtures also depends on the counterion decreasing in the sequence Ba2+ greater than Ca2+. These surface properties suggest how a lipid anion may function as an ionophore in the translocation of alkaline earth cations.

Estimation of surface area and counterion binding characteristics in fatty amine monolayers from desorption kinetics.

The surface area per molecule of an un-ionized fatty amine is very similar to the surface area per molecule of an un-ionized fatty acid. Surface area increases with ionization in both fatty amine and fatty acid films. However, fatty amino cations have much smaller surface areas than the corresponding fatty acid anions. Thus counterion binding is stronger with fatty amine cations than with fatty acid anions. Surface area data show that counterion binding affinities for fatty amine cations decrease in the strong field sequence Cl- greater than Br- greater than I- greater than SCN-. Furthermore, surface areas in the presence of the most strongly bound counterions, Cl- and Br-, increase significantly with an increase in subphase ionic strength. These data are consistent with the formation of strong ion-pair bonds and their disruption with an increase in ionic strength. Fatty amine cations desorb as micelles with much lower relative diffusion coefficients than the corresponding fatty acid anions. Furthermore, relative diffusion coefficients for fatty amine cations are strongly dependent on the specific cation. These data show that fatty amine cations form larger micelles when they desorb in the presence of strongly bound counterions. Anions enhance the solubility of a fatty acid anion in the sequence Cl- less than I- less than SCN-, which is characteristic of chaotropic anions that disrupt water structure.

Kinetics of the processes of desorption from fatty acid monolayers.

The surface area, A, of contracting fatty acid monolayers was measured as a function of time, t, at constant surface pressure. In the initial temporal phase, ln A was linear with radical t. In a subsequent steady-state phase, ln A was linear with t. The initial desorption coefficient for sodium palmitate, K(i), and the steady-state desorption coefficient, K(s), varied directly with surface pressure and subphase pH, and these desorption coefficients also varied with the composition of the subphase buffer. However, the K(s)/K(i) ratio was independent of these variables. The diffusion coefficient, D(25), for sodium palmitate calculated from desorption coefficient ratios was 4.8 +/- 0.6 x 10(-6) cm(2)/sec. This value was in reasonable agreement with D(25) for sodium palmitate measured by time-lag diffusion, 3.7 +/- 0.6 x 10(-6) cm(2)/sec. D(25) values obtained for a series of fatty acids suggested that higher members of the series diffused as small aggregates averaging two to four molecules in size. Kinetic and diffusion data both supported a model for the desorption process described by Ter Minassian-Saraga.

Surface areas of naturally occurring lipid classes and the quantitative microdetermination of lipids.

The surface area (A) of a lipid was directly proportional to the amount of lipid in a surface film (A = k micro moles) measured at constant surface pressure, temperature, and subphase composition. A surface area of 2300 cm(2)/ micro mole was obtained for cholesterol isolated from human adrenal and aorta and for cholesterol from hydrolysates of cholesteryl esters isolated from the same tissues. Unsaturated methyl esters that contained from one to four cis double bonds had the same surface area, 39.4 A(2)/molecule. As a consequence, naturally occurring triglyceride mixtures which had similar saturated-unsaturated fatty acid ratios had the same surface area, 6090 cm(2)/ micro mole. Naturally occurring phospholipid mixtures had the same surface area, 4590 cm(2)/ micro mole, and it appeared that the composition of these mixtures was regulated to control the physical properties of the mixtures. Surface area was much more sensitive than colorimetric procedures for the estimation of cholesterol and triglycerides. The surface area/molecule was a criterion of purity and an expanded surface area/molecule was an indication of autoxidation. Thus, surface area measurements were valuable for both the microdetermination and the characterization of lipid classes.

Effect of ionization and cation selectivity on the expansion of stearic acid monolayers.

Force-area isotherms of stearic acid and stearic acid-stearyl alcohol mixtures were investigated on alkaline subphases that contained Tris, Na(+), or K(+) cations and that varied in pH and ionic strength. The monolayer behaved as though ionization was effectively complete in the expanded region of the force-area isotherm. Surface pressure in this region was independent of pH and varied inversely with ionic strength as predicted by the Davies equation. The monolayer behaved as a partially ionized film in the plateau region of the force-area isotherm. Surface pressure in this region varied directly with pH and ionic strength as predicted by a modified Davies equation for partially ionized monolayers. The neutral molecule, stearyl alcohol, exerted a large condensing effect on the ionized film at pH 12.8, and this condensing effect also supported the concept that a partially ionized stearic acid film existed in the plateau region of the force-area isotherm. A greater binding affinity for Na(+) than for K(+) showed that the stearate anion surface behaved as a strong field at pH 10 and above, and a greater binding affinity for K(+) than for Na(+) showed that the stearate anion surface behaved as a weak field at pH 9. The weak field explained in part the anomalous binding affinity of the large Tris cation for the stearate monolayer at pH 9.