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[Hydrolysis of primary amide groups in Asn/Gln-containing peptides].

Peptides Boc-Ala-Asn/Gln-OH and Boc-Asn/Gln-Ala-OH were saponified with barium hydroxide to corresponding Asp/Glu-containing peptides. Under the conditions of saponification, Boc-Asn-Ala-OH additionally afforded Boc-Asp-OH, isopeptide Boc-Asp(Ala)-OH, and Boc-NHSuc > Ala-OH, with the third being the key intermediate in these transformations. Boc-Asp(OMe)-Ala-OMe underwent similar transformations under treatment with diazomethane or triethylamine. Saponification with barium hydroxide was accompanied by a high epimerization of N-terminal amino acid residues, whereas the products of the diazomethane treatment of Boc-Asp(OMe)-Ala-OMe had a low degree of epimerization.

Amides↗

Block of inwardly rectifying K+ currents by extracellular Mg2+ and Ba2+ in bovine pulmonary artery endothelial cells.

Using whole-cell patch clamp technique, we investigated the blocking effects of extracellular Ba2+ and Mg2+ on the inwardly rectifying K+ (KIR) currents of bovine pulmonary artery endothelial cells (BPAEC). The BPAEC KIR channel has recently been identified as Kir2.1 of the Kir2.0 subfamily. Block of KIR currents by Mg2+ (3-30 mM) was instantaneous, and increased with hyperpolarization slightly (Kd at -160 and 0 mV was 9.5 and 23.2 mM, respectively). The apparent fractional electrical distance (delta) of the Mg2+ binding site is calculated to be 0.07 from the outer mouth of the channel pore. Ba2+ (0.3-10 microM) time-dependently blocked the KIR currents with a much higher potency and stronger voltage-dependence (Kd at -160 and 0 mV was 1.0 and 41.6 microM, respectively). The Ba2+ binding site had a delta value of 0.34. Our data suggest that Mg2+ binds to a very superficial site of the KIR channel, while Ba2+ binds to a much deeper site, sensing much more of the membrane electric field. Thus, the BPAEC Kir2.1 appears to be pharmacologically different from the Kir2.1 reported before in bovine aortic endothelial cells (BAEC), which has 2 sites for Mg2+ block (a deep site in addition to a shallow one), and a superficial and low-sensitivity site for Ba2+ block.

Animals↗

Tamoxifen acutely relaxes coronary arteries by an endothelium-, nitric oxide-, and estrogen receptor-dependent mechanism.

In epidemiological studies tamoxifen has been associated with a reduction in the incidence of fatal myocardial infarction in women. However, the effects of tamoxifen on coronary artery reactivity are unknown. We hypothesized that tamoxifen would relax precontracted isolated rabbit coronary arteries. Rings of coronary artery from adult male and nonpregnant female New Zealand White rabbits were suspended in organ baths containing Krebs' solution; isometric tension was measured. Tamoxifen (0.1-100 microM) induced significant endothelium-dependent relaxation in precontracted rabbit coronary arteries. This relaxation was inhibited by N(omega)-nitro-L-arginine methyl ester and the estrogen receptor antagonist ICI 182,780. There was no significant effect on calcium concentration-dependent contraction curves. These data suggest that tamoxifen has beneficial effects on coronary reactivity that could, at least in part, account for the reduction in risk of fatal myocardial infarction in women taking tamoxifen.

Animals↗

Class V lesions restored with flowable composite and added surface sealing resin.

This in vitro study evaluates the marginal sealing ability of three different flowable composite systems at enamel and dentin margins, using sealants. Class V preparations were made on the facial surfaces in enamel and dentin/cement of 60 freshly extracted teeth, which were allotted randomly into six groups of ten. The control groups (I, II, and III) showed leakage in the majority of cases. However, Groups IV and V had very limited microleakage scores (2 of 30) in comparison to Group VI (p < 0.001, Mann-Whitney Test).

Barium Compounds↗

Microleakage at the cervical margin of composite Class II cavities with different restorative techniques.

This study evaluated the microleakage at the cervical margins of Class II composite resin restorations restored with different techniques. Slot cavities were prepared on both proximals of 40 unerupted third molars with one cervical margin located above and the other below the cementoenamel junction. The prepared teeth were randomly assigned to four groups and restored using the following techniques: (I) 3-Sited, (II) Directed-Shrinkage, (III) Resin-Modified Glass-Ionomer Cement or a (IV) Flowable Composite as the gingival increment. All restorations were placed with the same bonding agent and composite resin. The difference among the groups was on the first increment and on its insertion and polymerization techniques. After restoration, the teeth were thermocycled and exposed to a dye. Results showed that all the techniques worked well for enamel, with almost no leakage. However, on cementum, all techniques demonstrated moderate to severe leakage.

Barium Compounds↗

Thyrotropin-releasing hormone activates KCa channels in gastric smooth muscle cells via intracellular Ca2+ release.

Thyrotropin-releasing hormone (TRH) is released in high concentrations into gastric juice, but its direct effect on gastric smooth muscles has not been studied yet. We undertook studies on TRH effect on gastric smooth muscle using contraction and patch clamp methods. TRH was found to inhibit both acetylcholine- and BaCl2-induced contractions of gastric strips. TRH, applied to single cells, inhibited the voltage-dependent Ca2+ currents and activated the whole-cell K+ currents. The TRH-induced changes in K+ currents and membrane potential were effectively abolished by inhibitors of either intracellular Ca2+ release channels or phospholipase C. Neither activators, nor blockers of protein kinase C could affect the action of TRH on K+ currents. In conclusion, TRH activates K+ channels via inositol-1,4,5-trisphosphate-induced release of Ca2+ in the direction to the plasma membrane, which in turn leads to stimulation of the Ca2+-sensitive K+ conductance, membrane hyperpolarization and relaxation. The data imply that TRH may act physiologically as a local modulator of gastric smooth muscle tone.

Acetylcholine↗

Cholinergic modulation of stellate cells in the mammalian ventral cochlear nucleus.

The main source of excitation to the ventral cochlear nucleus (VCN) is from glutamatergic auditory nerve afferents, but the VCN is also innervated by two groups of cholinergic efferents from the ventral nucleus of the trapezoid body. One arises from collaterals of medial olivocochlear efferents, and the other arises from neurons that project solely to the VCN. This study examines the action of cholinergic inputs on stellate cells in the VCN. T stellate cells, which form one of the ascending auditory pathways to the inferior colliculus, and D stellate cells, which inhibit T stellate cells, are distinguished electrophysiologically. Whole-cell recordings from stellate cells in slices of the VCN of mice demonstrate that most T stellate cells are excited by cholinergic agonists through three types of receptors, whereas all D stellate cells tested were insensitive to cholinergic agonists. Nicotinic excitation in T stellate cells has two components. The faster component was blocked by alpha-bungarotoxin and methyllycaconitine, suggesting that receptors contained alpha7 subunits; the slower component was insensitive to both. Muscarinic receptors excite T stellate cells by blocking a voltage-insensitive, "leak" potassium conductance. Our results suggest that cholinergic efferent innervation enhances excitation by sounds of T stellate cells, opposing the inhibitory action of cholinergic innervation in the cochlea that is conveyed indirectly through the glutamatergic afferents. The inhibitory action of D stellate cells on their targets is probably not affected by cholinergic inputs. Excitation of T stellate cells by cholinergic efferents would be expected to enhance the encoding of spectral peaks in noise.

Acetylcholine↗

[Observation of antiarrhythmic effects of Cinnamomum migao H. W. Li on experimental arrhythmia].

OBJECTIVE: To investigate the effects of Cinnamomum migao on experimental arrhythmia. METHOD: Arrhythmic models of mice, rabbits, guinea pigs and rats were built using chloroform(Chl), adrenalin(Adr), strophanthin-K (Spt-K) and barium chloride (BaCl2). The affected animals were divided randomly into three groups: control group, Cinnamomum migao (CV-3) group and mexiletine (MXL) group, so as to observe and compare the antiarrhythmic effects. RESULT: CV-3 could reduce the incidence of ventricular fibrillation caused by ch1 in mice and the ventricular tachycardia induced by Adr in rabbits, delay the onset time of this arrhythmia, increase the arrhythmic doses of Spt-K in guinea pigs, reduce the incidence of some arrhythmia caused by BaCl2 in rats and slow down their heart rate. CONCLUSION: CV-3 has obvious antiarrhythmic effects on experimental arrhythmia. The mechanism of these effects is probably related to the arrest of the intraflow of Na+, Ca2+ in the cardiac cells and the depression of their cardiac autoarrhythmicity and conductivity.

Animals↗

Effect of dentin desensitizers and dentin bonding agents on dentin permeability.

PURPOSE: To determine the effect of dentin desensitizing agents (oxalates and glutaraldehyde) and dentin bonding agents, with and without resin-based composite (RBC) on dentin permeability and morphology of the dentin surface. The influence of saliva and toothbrushing on these agents was also evaluated. MATERIALS AND METHODS: Radicular dentin discs +/-1 mm thick were obtained from extracted human teeth. The thickness of the discs was standardized by grinding them with 600 grit silicon carbide powder. Both pulpal and periodontal surfaces of the discs were treated with 6% citric acid for 2 min prior to the baseline measurement of dentin permeability. Permeability was measured for the discs, using a split-chamber apparatus, at four different time points: at baseline, after treatment with each desensitizing modality, after 24 hrs of saliva immersion and after simulated toothbrushing equivalent to 3 weeks of normal brushing. The treatment modalities were: Sensodyne Dentin Desensitizer, Gluma Dentin Desensitizer, All-Bond DS (primers without etching), Etch+Primers (All-Bond 2 system), Etch+Primers+Adhesive and Etch+Primers+Adhesive+RBC (AElite Flo). Radicular dentin discs were prepared separately for scanning electron microscopy (SEM). One group was left untreated as the control group. Discs in the other groups were treated with the modalities listed above. One disc from each group was viewed under the SEM after treatment with the desensitizing modality. The second disc in each group was viewed after 24 hrs of saliva immersion following desensitizing treatment and the third disc after 3 weeks of simulated toothbrushing following desensitizing treatment and saliva immersion. RESULTS: The greatest reduction in permeability was caused by All-Bond DS followed by Sensodyne Dentin Desensitizer and Etch+Primers+Adhesive+RBC. Gluma Dentin Desensitizer, Etch+Primers, and Etch+Primers+Adhesive did not reduce permeability by a significant amount. Saliva immersion increased permeability significantly only for the Gluma and Etch+Primers+Adhesive groups. There was a trend towards increase in permeability after toothbrushing; however, this increase was not statistically significant for any group (P > 0.05). Thus, placement of primers without etching (All-Bond DS) reduced permeability more than any other treatment modality. These data indicate that etching a sensitive dentin area may be appropriate only if RBC is to be placed in the area. Exposure to saliva and toothbrushing countered the decrease in permeability caused by dedicated desensitizers and dentin bonding agents. The SEM micrographs revealed that in most cases, form corroborated function and the morphology of the dentin surface corresponded to the dentin permeability values.

Acid Etching, Dental↗

Effect of stepped light exposure on the volumetric polymerization shrinkage and bulk modulus of dental composites and an unfilled resin.

PURPOSE: To compare the volumetric polymerization shrinkage and dynamic bulk modulus of five resins and an experimental unfilled resin polymerized with a stepped power light and with a conventional one step curing light. MATERIALS AND METHODS: Parallel sided glass rings 5 mm in diameter and 2 mm high were etched for 5 min with hydrofluoric acid and silanated. The density and volume of the rings were calculated before they were filled with either SureFil, AEliteflo, Pertac II, Z100, or an experimental unfilled resin. When cured, the resin was bonded to the glass ring which produced a mold with a configuration C-factor of approximately 0.8. The resin was cured for 40 s on each side using either a one step curing light or a stepped power curing light. After curing, the specimens were weighed in air and in water to calculate their final density and volumetric shrinkage. Ten samples were made using each light system and for each resin (total of 100 samples). The bulk modulus was determined for each resin using an ultrasonic method, and the percentage filler contents were determined by thermogravimetric analysis. RESULTS: The mean +/- SD volumetric polymerization shrinkage values ranged from 3.10 +/- 1.19% for SureFil using the one step light to 12.35 +/- 1.38% for the unfilled resin using the stepped power curing light. Student-Newman-Keuls post hoc test separated the 10 different materials into three significant groups at P= 0.05. The unfilled resin exhibited the highest volumetric shrinkage followed by the flowable composite (AEliteflo). For all materials tested, there was no significant difference in the volumetric shrinkage values when the resin was cured with the one step or the stepped power curing light. There was no significant difference in the volumetric shrinkage values for the hybrid composites (Pertac II, Z100, and SureFil). The volumetric shrinkage values were highly sensitive to filler weight (P < 0.001). Except for Z100, there was no difference in the specific gravity values and velocities of the longitudinal and shear waves and bulk modulus when the one step and stepped power curing lights were used. The bulk modulus values ranged from 6.30 +/- 0.19 GPa for unfilled resin, to 17.10 +/- 1.09 GPa for Z100. Curing with the one step or the stepped curing light had no significant effect on the modulus values for any resin with the exception of Z100, P = 0.05. The bulk modulus values were highly sensitive to filler weight, and higher modulus values were associated with increased filler weight (P < 0.001). There was no significant difference between the bulk modulus of Pertac II and SureFil (P = 0.05).

Acid Etching, Dental↗

Microleakage of Class II resin-based composite restorations with flowable composite in the proximal box.

PURPOSE: To compare microleakage of Class II hybrid resin-based composite (RBC) restorations with and without low viscosity (flowable) RBC in the proximal box. MATERIALS AND METHODS: Conservative Class II cavity preparations were made in 50 extracted premolars randomly assigned to five groups (n = 10). OptiBond FL dentin bonding agent, and Prodigy hybrid RBC were used to restore all cavity preparations. Preparations in four of the five groups were restored with flowable RBC as the initial increment in the proximal box. The four flowable RBCs tested were: AEliteFlo, Revolution, UltraSeal XT and Flow-It. In the control group, the entire preparation was restored only with hybrid RBC. Specimens were stored in water at 37 degrees C for 24 hrs after restoration, thermocycled 1,000x, immersed in 2% basic fuchsin, sectioned and evaluated for microleakage. Microleakage was scored on a 0-4 scale. RESULTS: Non-parametric Kruskal-Wallis one-way ANOVA followed by Wilcoxon signed ranks test was used for statistical analysis. The use of any of the four flowable RBCs tested did not influence microleakage significantly (P > 0.05) in Class II RBC restorations.

Analysis of Variance↗

Fracture strength of fragment-bonded teeth. Effect of calcium hydroxide lining before bonding.

PURPOSE: To measure, in vitro the strength of intact teeth, fracture-bonded teeth, and fracture-bonded teeth bonded after a preliminary calcium hydroxide lining of the dentin part of the remaining tooth. MATERIALS AND METHODS: The study used 30 young, similarly aged sheep incisors. Twenty incisors were fractured 2.5 mm from the incisal edge and another 10 incisors remained non-fractured and served as control (Group 1). Group 2 comprised 10 fractured incisors, which after 3 weeks water storage were bonded by use of acid etching and a dentin bonding agent. In Group 3, the root part of 10 fractured incisors were lined with calcium hydroxide and kept in water for 3 weeks. Then the lining was removed with a hand instrument before bonding as above. The strength of the teeth in Groups 1-3 was measured and the mean fracture strengths representing the three groups were compared by Newman-Keuls' multiple range tests at a 5% level of significance. RESULTS: The mean fracture strength of the lined teeth (9.6 MPa) was significantly different from that of intact teeth (17.8 MPa) as well as from that of the bonded but unlined teeth (15.8 MPa). The latter two means were not significantly different. It is concluded that the use of calcium hydroxide on the fracture surfaces of fractured teeth before bonding lowers the fracture strength of the restored teeth.

Acid Etching, Dental↗

Evaluation of barium hydroxide treatment efficacy on a dolomitic marble.

The Arch of Peace, by Luigi Cagnola, is one of the most famous neoclassical monuments in Milan. It has been subjected to conservative intervention in 1998. In the present paper the efficacy of the consolidation by means of barium hydroxide has been evaluated. The stone material showed severe degradation phenomena as: erosion, pulverisation, exfoliation. The analytical data acquired through X-ray diffraction (XRD), infrared spectrophotometry (FTIR) and scanning electron microscopy (SEM-EDX), allowed to compare the conditions of stone before and after the treatment with barium hydroxide. The presence of barium has been put in evidence mainly on the surface as barium sulphate, whereas barium is only sporadically present within the thickness of the decayed material. The treatment was judged not satisfying and its inefficacy is, most probably, due to a not suitable cleaning procedure carried out before the consolidation.

Barium Compounds↗

Effect of cyclical lateral forces on microleakage in cervical resin composite restorations.

OBJECTIVE: The aim of this study was to assess the effect on microleakage of cyclic lateral fatigue forces in different types of cervical resin composite restorations. METHOD AND MATERIALS: V-notch cavities were prepared at the cementoenamel junction in 60 extracted human premolars and restored with a bonding system along with one of the following resin composites: hybrid resin composite, microfilled resin composite, and flowable resin composite (20 each). Ten specimens from each group of 20 were subjected to 8,400 cycles of lateral fatigue forces (44 N) at the occlusal half of the clinical crown in a machine specifically developed for this purpose. The other 10 specimens of each group served as controls, representing the microleakage that occurred without lateral fatigue forces. All specimens were stained with a 50% silver nitrate solution and sectioned sagittally. A scoring system of 0 to 6 was used to record the amount of die penetration along the restoration-tooth interface. RESULTS: There was leakage in both the fatigued specimens and the nonfatigued control specimens. There were significant differences in the mean values of microleakage for the restorative materials studied. In the nonfatigued groups, hybrid resin specimens exhibited significantly more microleakage at the dentin-resin interface than did microfilled or flowable resin specimens. In the fatigued groups, hybrid resin specimens exhibited significantly more leakage at the dentin-resin interface than did microfilled resin specimens. CONCLUSION: The least elastic resin composite tested, the hybrid material, generally showed the most leakage. This suggests that the elasticity of these resin composite restorative materials may be a factor in the amount of microleakage in cervical restorations.

Barium Compounds↗

Flow characteristics and film thickness of flowable resin composites.

Flowable resin composites have been recommended for many clinical uses and have been formulated in a variety of compositions and viscosities to meet various uses. This study compared the variation in viscosity of flowable resin composites using the ADA Flow Test and measured film thickness with a test to simulate flow during cementation. The film thickness test for the flowable resin composites was performed at three different seating pressures to simulate pressure variation during seating of porcelain veneers, one of the potential uses of flowable resin composites that may favor a lower viscosity. The following flowable resin composites were evaluated: Revolution, StarFlow, Aeliteflo LV, Aelite, Flow-It, FloRestore, Versaflo, Durafill Flow and Tetric Flow, with Nexus2, a composite luting resin used as a control. Flow characteristic measurements suggest that resin composites may be divided into high flow (StarFlow, Revolution, Aeliteflo LV), medium flow (FloRestore, Durafill Flow, Flow-It) and low flow (Tetric Flow, Versaflo, Nexus2, Aelite) groupings. The film thickness measurements agreed with the ADA flow test, except for two exceptions. Durafill Flow resin composite had a higher film thickness than expected based on the ADA flow test. Also, Aeliteflo unexpectedly had a lower film thickness. At the lowest seating pressure (.016MPa) tested, eight out of the nine resin composites tested as well as the control luting resin had a film thickness greater than the 25 microns used as the clinical standard for cement film thickness. However, at the highest seating pressure (.038 MPa) tested, only two and the control, Versaflo, Durafill Flow and Nexus2, had film thicknesses significantly greater than 25 microns. The highly filled Nexus2 luting resin had the highest film thickness at all seating pressures.

Analysis of Variance↗

Effect of interfacial bond quality on the direction of polymerization shrinkage flow in resin composite restorations.

Shrinkage in light curing resin composites is assumed to be directed toward the light source. However, the strong bond at the dentin-resin interface achieved by newer generation dentin bonding systems may affect the direction of polymerization shrinkage. In this study, various curing modes of adhesive resin simulating different bond qualities were applied to determine the extent of interfacial gap formation with a scanning electron microscope. We also measured the free surface depression with a profilometer. The direction of polymerization shrinkage was inferred from the ratio of the interfacial gap measurement at the floor to the free surface depression. Various curing modes used in this study include Group 1: light curing of resin composite without the bonding agent as the negative control; Group 2: simultaneous light curing of the bonding agent and resin composite; Group 3: start of the chemical cure of the dual-cured bonding agent before light curing the resin composite; Group 4: curing the light-initiated bonding agent before insertion and light curing of the resin composite. When the bonding agent was light cured prior to inserting the resin composite (Group 4), the free surface depression was the greatest and the interfacial gap smallest among those in all groups. Therefore, if a good bond between dentin and resin composite can be established, the shrinkage flow will be directed toward a center located near the bonded interface rather than toward the incident light, thus reducing detrimental shrinkage stress.

Acid Etching, Dental↗

Calcium oscillations in freshly isolated neonatal rat cortical neurons.

AIM: To investigate the mechanisms of intracellular calcium concentration ([Ca2+]i) oscillations in freshly isolated neonatal rat cortical neurons. METHODS: Cortical neurons were isolated from neonatal rats 6-7 d after birth by enzymatic digestion. [Ca2+]i changes were measured in a microscopic calcium measurement system with Fura-2 as indicator. RESULTS: In a total of 82 neurons recorded, 47 showed spontaneous [Ca2+]i oscillations. The spontaneous [Ca2+]i oscillations were dependent on [Ca2+]o. Removal of [Ca2+]o completely abolished spontaneous oscillations. Tetraethylammonium 1 mmol/L increased both the amplitude and frequency of calcium oscillations, whereas the frequency was increased by Cs+ 1 mmol/L. Ba2+ 1 mmol/L, in contrast, induced [Ca2+]i oscillations superimposed on a sustained phasic increase. CONCLUSION: Spontaneous [Ca2+]i oscillation is an intrinsic property of neonatal rat cortical neurons. Potassium channels play an important role in the control of both the amplitude and frequency of [Ca2+]i oscillations in cortical neurons.

Animals↗