Effect of whitening products on sound enamel and on artificial caries lesions during a cariogenic challenge.

OBJECTIVE: To investigate the effect of fluoride-containing whitening products on sound enamel and on artificial caries lesions during a cariogenic challenge. MATERIALS AND METHODS: Bovine enamel specimens (n = 120) with three areas [non-treated sound enamel (NSE), treated sound enamel (TSE), and treated artificial caries lesion (TACL)] were randomly assigned to the four groups: whitening mouthrinse (WM: 2.5% hydrogen peroxide-100 ppm F-), placebo mouthrinse (PM: 0% hydrogen peroxide-100 ppm F-), whitening gel (WG: 10% carbamide peroxide-1130 ppm F-), and deionized water (negative control; NC). The treatments (2 min for WM, PM, and NC, and 2 h for WG) were carried out during a 28-day pH-cycling model (6 × 60 min demineralization/day). Relative surface reflection intensity (rSRI) and transversal microradiography (TMR) analyses were performed. Fluoride uptake (surface and subsurface) was measured in additional enamel specimens. RESULTS: For TSE, a higher value of rSRI was observed in WM (89.99% ± 6.94), and a greater decrease in rSRI was observed for WG and NC, and no sign of mineral loss was verified for all groups (p > 0.05). For TACL, rSRI significantly decreased after pH-cycling for all experimental groups with no difference between them (p < 0.05). Higher amounts of fluoride were found in WG. WG and WM exhibited intermediate values of mineral loss, similar to PM. CONCLUSIONS: The whitening products did not potentialize the enamel demineralization under a severe cariogenic challenge, and they did not exacerbate mineral loss of the artificial caries lesions. CLINICAL RELEVANCE: Low concentrated hydrogen peroxide whitening gel and mouthrinse containing fluoride do not intensify the progression of caries lesions.

Optical Property Stability of Light-Cured versus Precured CAD-CAM Composites.

Objective: The aim of this study was to evaluate the optical property changes after staining of precured (PC) and light-cured (LC) composites. Materials and Methods: Specimens were prepared using different LC composites (GrandioSO-Voco, Filtek Z350-3M/ESPE, Opallis-FGM, and Kalore-GC) and four PC blocks (Grandio Blocs-Voco, Lava Ultimate-3M ESPE, Brava Block-FGM, and Cerasmart-GC) from the same manufacturers (n = 20). Baseline color, gloss, translucency, and fluorescence were evaluated. The staining protocol was performed for 15 days, and the final optical properties were reevaluated. Results: The changes in each property were calculated (ΔGloss, ΔTranslucency, ΔFluorescency, ΔE ∗ 00). Data were analyzed by ANOVA and Tukey's test (α = 5%). Changes in all properties were observed after staining for all materials, with darkening and reduction of gloss, fluorescence, and translucency. Nonsignificant differences were observed between the light-cured and precured materials of the same manufacturer for ΔG and ΔT, but significant differences existed for ΔF and ΔE ∗ 00. For ΔF, the only significant differences were observed between Brava Block and Opallis (smaller). For ΔE ∗ 00, only the light-cured composites GrandioSO and Z350 showed significantly less change than the corresponding blocks. Precured composites were affected the same way as light-cured ones by the staining in relation to the reduction of gloss and translucency. Conclusion: A higher reduction in fluorescence was observed for only one brand of block and was similar for the others. The two brands of light-cured materials showed less staining, while for the others, the staining was similar. The effects of staining vary according to the composite formulation.

Toothbrushing abrasion susceptibility of enamel and dentin bleached with calcium-supplemented hydrogen peroxide gel.

UNLABELLED: The objective of this study was to evaluate enamel and dentin susceptibility to toothbrushing abrasion, after bleaching with 7.5% hydrogen peroxide (HP) gel supplemented or not with 0.5% calcium gluconate (Ca). Toothbrushing was performed immediately and 1h after bleaching, with two suspensions (high and low abrasivity). Bovine enamel and dentin specimens were divided into 12 groups (n=10) according to the bleaching gel (with and without Ca), slurry abrasivity (high or low) and elapsed time after bleaching (immediately and after 1h). As control, a group was not bleached, but abraded. The treatment cycle (7 d) consisted of bleaching (1h) and toothbrushing (135 strokes/day) immediatelly or after 1h of artificial saliva exposure. Surface roughness and surface loss (µm) were measured by profilometry and analysed by three-way ANOVA (5%). Surface roughness means were significantly influenced by slurry abrasivity (p<0.0001). For enamel loss, significant triple interaction was observed (p<0.0001). HP-bleached groups and immediately brushed with high-abrasive slurry exhibited increased loss (1.41±0.14) compared to other groups (µm). Control and HP+Ca-bleached groups brushed after 1h with low abrasive slurry presented the lowest loss (0.21±0.03/0.27±0.02). For dentin loss, significant interaction was observed for bleaching and interval factors (p<0.001). 7.5%HP-bleached groups and immediately brushed showed significantly higher loss (8.71±2.45) than the other groups. It was concluded that surface roughness increased when high abrasive was used, independently of bleaching. 7.5%HP increased enamel and dentin loss, mainly with high abrasive slurries. Calcium supplementation of bleaching gel reduced surface loss. Additionally, in order to minimize tooth wear susceptibility, it is recommended to delay brushing after bleaching. CLINICAL RELEVANCE: After bleaching gel application, postponing toothbrushing is recommended, as well as brushing with low abrasive dentifrices. Additionally, supplementation of hydrogen peroxide gel with calcium-based remineralizing agent potentially reduces tooth loss after abrasion.

Longitudinal Evaluation of Bond Strength to Enamel of Dental Adhesive Systems Associated with Nd:YAG Laser.

OBJECTIVES: This study evaluated the durability of bond strength to enamel using total-etch (Single Bond/SB) and self-etch (Clearfil SE Bond/CSEB) adhesives associated with neodymium:yttrium-aluminu-garnet (Nd:YAG) laser irradiation through the uncured adhesives. METHODS: Bovine incisors were worn to expose an area of enamel and were divided into four groups: group 1 (control) SB + polymerization; group 2 (control) CSEB + polymerization; group 3 (laser) - SB + Nd:YAG laser (174.16 J/cm(2)) + polymerization; and group 4 (laser) CSEB + Nd:YAG (174.16 J/cm(2)) + polymerization. Blocks of composite were fabricated and stored for 24 hours or 12 months, sectioned into beams, and submitted to microtensile tests. Results were analyzed by three-way analysis of variance (ANOVA) (adhesive, technique, and storage time) and Tukey tests. RESULTS: ANOVA revealed significant differences for adhesive × technique and technique × storage time (p<0.05). The mean values (MPa) for interaction adhesive × technique (standard deviation) were as follows: SB/control = 35.78 (6.04)a; SB/laser = 26.40 (7.25)b, CSEB/control = 26.32 (5.71)b, CSEB/laser = 23.90 (7.49)b. For interaction technique × storage time the mean values were as follows: control/24 hours = 32.58 (6.49)a; control/12 months = 29.52 (8.38)a; laser/24 hours = 29.37 (5.71)a; laser/12 months = 20.92 (6.5)b. Groups with the same letters showed no statistically significant differences. CONCLUSION: Scanning electron microscope analysis showed evident areas of micromorphological alterations in lased samples after 12 months of water storage. Nd:YAG laser irradiation of enamel through unpolymerized total-etch adhesive significantly reduced bond strength compared with the control. Bond strength decreased when enamel samples irradiated with Nd:YAG laser through unpolymerized adhesives were stored in water for 12 months.

Effect of hydrogen peroxide concentration on enamel color and microhardness.

OBJECTIVES: The aim of this study was to investigate the effect of hydrogen peroxide gels with different concentrations (20%, 25%, 30%, and 35%) on enamel Knoop microhardness (KNH) as well as on changes in dental color (C). METHODS: Cylindrical specimens of enamel/dentin (3-mm diameter and 2-mm thickness) were obtained from bovine incisors and randomly divided into six groups (n=20), according to the concentration of the whitening gel (20%, 25%, 30%, 35%, control, thickener). After polishing, initial values of KNH0 and color measurement, assessed by spectrophotometry using the CIE L*a*b* system, were taken from the enamel surface. The gels were applied on the enamel surface for 30 minutes, and immediate values of KNHi were taken. After seven days of being stored in artificial saliva, new measures of KNH7 and color (L7* a7* b7*, for calculating ΔE, ΔL, and Δb) were made. Data were submitted to statistical analysis of variance, followed by Tukey test (p<0.05). RESULTS: Differences in gel concentration and time did not influence the microhardness (p=0.54 and p=0.29, respectively). In relation to color changes, ΔE data showed that the 35% gel presented a higher color alteration than the 20% gel did (p=0.006). CONCLUSION: Bleaching with 35% hydrogen peroxide gel was more effective than with the 20% gel, without promoting significant adverse effects on enamel surface microhardness.

Color masking of developmental enamel defects: a case series.

Developmental defects involving color alteration of enamel frequently compromise the esthetic appearance of the tooth. The resin infiltration technique represents an alternative treatment for color masking of these lesions and uniformization of tooth color. This technique is considered relatively simple and microinvasive, since only a minimal portion of enamel is removed. This article illustrates the color-masking effect with resin infiltration of fluorosis and traumatic hypomineralization lesions with a case series. The final esthetic outcomes demonstrated the ability of the resin infiltrant to mask the color of white developmental defect lesions, resulting in satisfactory clinical esthetic improvements. However, in more severe cases, the color-masking effect was not complete.

Influence of pH on the effectiveness of hydrogen peroxide whitening.

OBJECTIVE: To evaluate the influence of pH on the bleaching effect of hydrogen peroxide on chromogen agents. METHOD: Hydrogen peroxide 50% was mixed with red wine or with an alcoholic solution of tobacco in glass cuvettes, resulting in final peroxide concentrations of 16.97% and 21.12%, respectively. The pH of this mixture was measured and adjusted with 3.3 M HCl solution or 2.5 M NaOH solution to obtain the final pH values of 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, and 9.0. After mixing, the color of these solutions was evaluated in a reflectance spectrophotometer; readings were repeated after 10 minutes for the wine solution and 20 minutes for the tobacco solution. Ten samples were prepared for each solution at each pH. Color changes (Delta E) were calculated. The data were statistically analyzed using analysis of variance one-way and Tukey tests, with a significance level of 5%. RESULTS: There were significant differences among the different pH values for the wine and tobacco solutions (p=0.0001). The Tukey test showed that for both solutions, pH 9.0 resulted in a significantly greater bleaching effect than the other values tested. CONCLUSION: The efficacy of hydrogen peroxide bleaching is directly proportional to the increase in its pH.

The effect of three desensitizing agents on dentin hypersensitivity: a randomized, split-mouth clinical trial.

The aim of this study was to evaluate the efficacy of three desensitizing agents to provide relief to dentin hypersensitivity after one session in a four-week follow-up. Forty selected patients participated in a double-blind study following a split-mouth model. One application of the desensitizing agents (A, Admira Protect [Voco]; B, Bifluorid 12 [Voco]; and C, Colgate Pro-Relief in office [Colgate Palmolive]) was performed in three different quadrants for each patient. Each tooth was evaluated by tactile and evaporative stimuli, and the sensitivity response was measured using the Visual Analogue Scale. Evaluations were performed at baseline, immediately after treatment, and after one, two, three, and four weeks. The application of Kruskal-Wallis and Dunn multiple comparisons tests (5%) for both tactile and evaporative stimuli showed that all agents presented a significant desensitizing effect. In groups A and B this relief was maintained for four and three weeks, respectively, as measured by tactile stimulus and for four weeks with evaporative stimulus. The desensitizing effect for group C was maintained for two weeks for both tactile and evaporative stimuli. It is concluded that all desensitizing agents tested were effective in reducing sensitivity compared to baseline values. One application of Admira Protect and Bifluorid 12 presented a longer-lasting desensitizing effect than did Colgate Pro-Relief (applied in the office) on both tactile and evaporative stimuli.

Ultrasonic activation of internal bleaching agents.

AIM: To evaluate the effectiveness of ultrasonic activation of bleaching agents during ex vivo internal bleaching. METHODOLOGY: Fifty canine human teeth were artificially stained, root filled and divided into five groups (n = 10) that received SP - sodium perborate plus deionized water (control group), CP - 37% carbamide peroxide gel, CPUS - 37% carbamide peroxide gel plus ultrasonic application, HP - 35% hydrogen peroxide gel or HPUS - 35% hydrogen peroxide gel plus ultrasonic application. In groups CP and HP, the bleaching agent was left inside the pulp chamber for three applications of 10 min. In groups CPUS and HPUS, the same process was performed, but ultrasonic vibration was applied to the bleaching agent by an alloy tip for 30 s, with 30 s intervals. Two sessions were performed. The colour was measured initially and after each session by an intraoral dental spectrophotometer. The variation (Δ) of the colour parameters based on the CIELab system L*, a* and b*, and the colour alteration ΔE* were calculated after first and second section. Data were analysed by one-way anova and Tukey's test. RESULTS: There was no significant difference amongst groups for ΔL*, Δa* and ΔE*, but there was a significant difference for Δb* in the first and second sessions (P = 0.0006 and 0.0016, respectively). After the first session, Δb* was significantly greater for groups HP and HPUS, without a significant difference between them. For the second session, group HPUS had the greatest Δb* values, but they were similar to groups HP and SP; group CP had the lowest values, which were similar to groups CPUS and SP. CONCLUSION: Ultrasonic activation of bleaching agents during ex vivo internal bleaching was no more effective than conventional internal bleaching procedures, without activation.

Efficacy of mouth rinses and toothpaste on tooth whitening.

OBJECTIVES: People increasingly desire tooth whitening. Considering the wide range of whitening products on the market, this study evaluated the efficacy of whitening toothpastes and mouth rinses compared with the 10% carbamide peroxide (CP) whitening gel. METHODS: We obtained 120 cylindrical specimens from bovine teeth, which were darkened for 24 hours in a coffee solution. The color measurement was performed by a spectrophotometer using the CIE L*a*b* system, and specimens were divided into six groups according to the use of the following agents: group 1, conventional fluoridated toothpaste; group 2, Close Up White Now; group 3, Listerine Whitening; group 4, Colgate Plax Whitening; group 5, experimental mouth rinse with Plasdone; and group 6, 10% CP Whiteness Perfect. After the simulation of 12 weeks of treatment for groups 1 to 5 and 14 days of treatment for group 6, the specimens were subjected to a new color reading. RESULTS: Data were subjected to one-way analysis of variance (α=0.05), which showed significant differences among groups after 12 weeks for ΔE (p=0.001). Results of the Tukey test revealed that groups 3, 4, and 6 presented significantly higher color alteration than groups 1, 2, and 5. CONCLUSIONS: The whitening toothpaste Close Up White Now and the experimental mouth rinse with Plasdone showed similar color alteration as conventional toothpaste after a 12-week treatment simulation. These groups presented significantly lower color alteration compared with whitening mouth rinses Listerine and Colgate Plax Whitening, which showed similar results to those observed after 14 days of bleaching with 10% CP treatment.

Two-year clinical performance of self-etching adhesive systems in composite restorations of anterior teeth.

OBJECTIVE: The aim of this study was to evaluate the two-year clinical performance of Class III, IV, and V composite restorations using a two-step etch-and-rinse adhesive system (2-ERA) and three one-step self-etching adhesive systems (1-SEAs). MATERIAL AND METHODS: Two hundred Class III, IV, and V composite restorations were placed into 50 patients. Each patient received four composite restorations (Amaris, Voco), and these restorations were bonded with one of three 1-SEAs (Futurabond M, Voco; Clearfil S3 Bond, Kuraray; and Optibond All-in-One, Kerr) or one 2-ERA (Adper Single Bond 2/3M ESPE). The four adhesive systems were evaluated at baseline and after 24 months using the following criteria: restoration retention, marginal integrity, marginal discoloration, caries occurrence, postoperative sensitivity and preservation of tooth vitality. After two years, 162 restorations were evaluated in 41 patients. Data were analyzed using the χ(2) test (p<0.05). RESULTS: There were no statistically significant differences between the 2-ERA and the 1-SEAs regarding the evaluated parameters (p>0.05). CONCLUSION: The 1-SEAs showed good clinical performance at the end of 24 months.

Influence of Nd:YAG or Er:YAG laser surface treatment on microtensile bond strength of indirect resin composites to resin cement. Lasers surface treatment of indirect resin composites.

This study evaluated the influence of the surface pretreatment of indirect resin composite (Signum, Admira Lab and Sinfony) on the microtensile bond strength of a resin cement. Sixty samples made of each brand were divided into 6 groups, according to surface treatment: (1) control; (2) controlled-air abrasion with Al2O3; (3) Er:YAG Laser 200 mJ, 10 Hz, for 10s; (4) Er: YAG Laser 300 mJ, 10 Hz, for 10 s; (5) Nd:YAG 80 mJ, S15Hz for 1 min; (6) Nd:YAG 120mJ, 15 Hz for 1 min. After treatments, all the groups received an application of 37% phosphoric acid and adhesive. The pair of blocks of the same brand were cemented to each other with dual resin cement. The blocks were sectioned to obtain resin-resin sticks (1 x1 mm) and analyzed by microtensile bond testing. The bond strength values were statistically different, irrespective of the surface treatment performed, with highest values for Sinfony (43.81 MPa) and lowest values for Signum (32.33 MPA). The groups treated with the Nd:YAG laser showed the lowest bond strength values and power did not interfere in the results, both for Nd:YAG laser and Er:YAG. Controlled-air abrasion with Al203 is an efficient surface treatment method and the use of the Nd:YAG and Er:YAG lasers reduced bond strength, irrespective of the intensity of energy used.

Influence of hydrogen peroxide bleaching gels on color, opacity, and fluorescence of composite resins.

The aim of the present study was to evaluate the effect of 20% and 35% hydrogen peroxide bleaching gels on the color, opacity, and fluorescence of composite resins. Seven composite resin brands were tested and 30 specimens, 3-mm in diameter and 2-mm thick, of each material were fabricated, for a total of 210 specimens. The specimens of each tested material were divided into three subgroups (n=10) according to the bleaching therapy tested: 20% hydrogen peroxide gel, 35% hydroxide peroxide gel, and the control group. The baseline color, opacity, and fluorescence were assessed by spectrophotometry. Four 30-minute bleaching gel applications, two hours in total, were performed. The control group did not receive bleaching treatment and was stored in deionized water. Final assessments were performed, and data were analyzed by two-way analysis of variance and Tukey tests (p<0.05). Color changes were significant for different tested bleaching therapies (p<0.0001), with the greatest color change observed for 35% hydrogen peroxide gel. No difference in opacity was detected for all analyzed parameters. Fluorescence changes were influenced by composite resin brand (p<0.0001) and bleaching therapy (p=0.0016) used. No significant differences in fluorescence between different bleaching gel concentrations were detected by Tukey test. The greatest fluorescence alteration was detected on the brand Z350. It was concluded that 35% hydrogen peroxide bleaching gel generated the greatest color change among all evaluated materials. No statistical opacity changes were detected for all tested variables, and significant fluorescence changes were dependent on the material and bleaching therapy, regardless of the gel concentration.

Effect of caries infiltration technique and fluoride therapy on microhardness of enamel carious lesions.

Enamel white spot subsurface lesions compromise esthetics and precede cavitation; therefore, they must be halted. The aim of this study was to evaluate the effect of a caries infiltration technique and fluoride therapy on the microhardness of enamel carious lesions. Subsurface carious lesions were produced in 60 bovine specimens with polished enamel surfaces. The specimens were divided into four groups (n=15), according to the treatment used: CON, control-immersion in artificial saliva; DF, daily 0.05% fluoride solution; WF, weekly 2% fluoride gel; and IC, resin infiltration (Icon). The specimens were kept in artificial saliva and evaluated for microhardness at five points: baseline, after caries production, after four and eight weeks of treatment, and a final evaluation after being submitted to a new acid challenge. The repeated-measures analysis of variance showed significant differences according to the type of treatment (TREAT; p=0.001) and time of evaluation (EV; p=0.001). The results of the Tukey test were TREAT: CON = 45.18 (±29.17)a, DF = 107.75 (±67.38)b, WF = 83.25 (±51.17)c, and IC = 160.83 (±91.11)d. Analysis of correlation between the TREAT and EV factors showed no significant differences for DF (138.63 ± 38.94) and IC (160.99 ± 46.13) after the new acid challenge. The microhardness results in decreasing order after eight weeks were IC > DF > WF > CON. It was concluded that the microhardness of carious lesions increased with the infiltration of resin, while the final microhardness after a new acid challenge was similar for DF and IC.

Tribulations and achievements: the early history of Olympism in Argentina.

By the end of the nineteenth century, modern sport had enchanted the people of Argentina. At that time the nation enjoyed a remarkable degree of economic prosperity and embarked on increasing political democratization. These circumstances, along with the fact that the nation was represented from the beginning, in 1894, on the International Olympic Committee seemed to favour Argentina as the spearhead of the diffusion of Olympism throughout South America. However, the country only enjoyed its first official Olympic participation in the Paris Games of 1924 - a few months after the establishment of the Argentine Olympic Committee. This essay explores the reception and diffusion of Olympism in Argentina. It reveals a process of gradual adoption including conflicting views on the relationship between the state and sport, several attempts at institutionalization, international misunderstandings and the role of politics and class.

Inhibition of the ecto-ATPdiphosphohydrolase of Schistosoma mansoni by thapsigargin.

ATPdiphosphohydrolases (ATPDases) are ubiquitous enzymes capable of hydrolyzing nucleoside di- and triphosphates. Although a number of possible physiological roles have been proposed for ATPDases, detailed studies on structure-function relationships have generally been hampered by the lack of specific inhibitors of these enzymes. We have previously characterized a Ca2+-activated ATPDase on the external surface of the tegument of Schistosoma mansoni, the etiologic agent of human schistosomiasis. In the present work, we have examined the effects of thapsigargin, a sesquiterpene lactone known as a high affinity inhibitor of sarco-endoplasmic reticulum calcium transport (SERCA) ATPase, on ATPDase activity. Whereas other lactones tested had little or no inhibitory action, thapsigargin inhibited ATP hydrolysis by the ATPDase (K(i) approximately 20 microM). Interestingly, hydrolysis of ADP was not inhibited by thapsigargin. The lack of inhibition of ATPase activity by orthovanadate, a specific inhibitor of P-type ATPases, and the inhibition of the Mg2+-stimulated ATP hydrolysis by thapsigargin ruled out the possibility that the observed inhibition of the ATPDase by thapsigargin could be due to the presence of contaminating SERCA ATPases in our preparation. Kinetic analysis indicated that a single active site in the ATPDase is responsible for hydrolysis of both ATP and ADP. Thapsigargin caused changes in both Vmax and Km for ATP, indicating a mixed type of inhibition. Inhibition by thapsigargin was little or not affected by changes in free Ca2+ or Mg2+ concentrations. These results suggest that interaction of thapsigargin with the S. mansoni ATPDase prevents binding of ATP or its hydrolysis at the active site, while ADP can still undergo catalysis.

Divalent cation dependence and inhibition of Schistosoma mansoni ATP diphosphohydrolase by fluorosulfonylbenzoyl adenosine.

Hydrolysis of ATP or ADP catalyzed by the ATP diphosphohydrolase of Schistosoma mansoni tegument was measured in the presence of different cations. ATP diphosphohydrolase was stimulated by micromolar concentrations of either Ca2+ or Mg2+, Ca2+ producing threefold higher maximal activities than Mg2+. Kinetic studies indicated that Ca2+ and Mg2+ compete for the same binding site on the enzyme. The effect of covalent labeling of ATP diphosphohydrolase with the ATP analog fluorosulfonylbenzoyl adenosine (FSO2BzAdo) was studied. Schistosome tegument was passed through with Sephadex G-50 filtration centrifugation columns to remove endogenous nucleotides, and this was followed by labeling with FSO2BzAdo. Incubation of ATP diphosphohydrolase with 1 mM FSO2BzAdo for 1 h inhibited ATPase or ADPase activities by 60% and 50%, respectively. Addition of ATP together with FSO2BzAdo provided greater than 90% protection against FSO2BzAdo inactivation, indicating that FSO2BzAdo binds to an ATP-binding site on the ATP diphosphohydrolase. Furthermore, addition of FSO2BzAdo to a medium containing intact worms caused 30% and 50% inhibition of ATPase and ADPase activities, respectively, indicating that the ATP-binding site of diphosphohydrolase is accessible to FSO2BzAdo from the external surface of S. mansoni worms.

In vitro development to blastocysts of early porcine embryos produced in vivo or in vitro.

The objective of this study was to compare the development of porcine embryos from the 2- and 4-cell stages to the blastocyst stage after in vivo or in vitro fertilization and in vivo or in vitro culture. Early-stage embryos were collected either from superovulated gilts 36 h after the second mating or after in vitro fertilization (IVF) of in vivo-matured oocytes, both followed by in vitro culture to the blastocyst stage. Blastocysts collected from superovulated donors served as controls. In the first experiment, a total of 821 2- and 4-cell embryos derived from in vivo-fertilized oocytes was cultured either in medium NCSU 23, modified Whittens' medium or modified KRB for 5 d. Significantly (P < 0.05 and P < 0.001) more embryos overcame the 4-cell block and developed to the blastocyst stage in medium NCSU 23 than in the 2 other culture media. Hatching was only observed in medium NCSU 23. In the second experiment, embryos derived from in vivo-matured oocytes fertilized in vitro were cultured in medium NCSU 23. Of 1869 mature oocytes 781 (41.8%) cleaved within 48 h after in vitro fertilization. A total of 715 embryos was cultured to the morula and blastocyst stages, and 410 (57.3%) overcame the developmental block stage, with 358 embryos (50.1%) developing to the morula and blastocyst stages. None of the embryos hatched, and the number of nuclei was significantly (P < 0.05) lower compared with that of in vivo-fertilized embryos (18.9 +/- 9.8 vs 31.2 +/- 5.8). In the third experiment, 156 blastocysts derived from in vitro fertilization and 276 blastocysts derived from in vivo fertilization and in vitro culture were transferred into synchronized recipients, while 164 blastocysts were transferred immediately after collection into 6 recipients, resulting in a pregnancy rate of 83.3%, with 35 piglets (on average 7.0) born. From the in vitro-cultured embryos, 58.3% (7/12) of the recipients remained pregnant at Day 35 after transfer, but only 33.3% maintained pregnancy to term, and 14 piglets (on average 3.5) were born. In contrast, the transfer of embryos derived from in vitro-fertilized oocytes did not result in pregnancies. It is concluded that 1) NCSU 23 is superior to modified Whittens' medium and modified KRB and 2) blastocysts derived from in vitro fertilization have reduced viability as indicated by the lower number of nuclei and failure to induce pregnancy upon transfer into recipients.

Erythrocytes contain cytoplasmic glycoproteins. O-linked GlcNAc on Band 4.1.

Previously we reported that the novel protein-saccharide linkage, O-linked N-acetylglucosamine (GlcNAc), is found in abundance on proteins associated with the cytoplasmic and nucleoplasmic faces of the nuclear pore complex. Here we demonstrate that O-GlcNAc moieties are also added to human erythrocyte cytoplasmic proteins. Intact or permeabilized erythrocytes, as well as subcellular fractions, were labeled with bovine milk galactosyltransferase and UDP-[3H] galactose. The proportion of the incorporated label found on O-GlcNAc was determined by a variety of chemical and enzymatic techniques. The bulk of the O-GlcNAc residues are found in the cytoplasm of erythrocytes, the majority of which are on an as yet unidentified 65-kDa protein. In addition, we have determined that Band 4.1, a protein which serves as a bridge joining the cytoskeleton to the inner surface of the plasma membrane in erythrocytes, also contains O-GlcNAc moieties. One of the sites of O-GlcNAc addition has been localized to the last 117 amino acids of the carboxy terminus of Band 4.1.