In vitro digestibility of beta-casein and beta-lactoglobulin under simulated human gastric and duodenal conditions: a multi-laboratory evaluation.

Initially the resistance to digestion of two cow's milk allergens, beta-casein, and beta-lactoglobulin (beta-Lg), was compared using a "high-protease assay" and a "low-protease assay" in a single laboratory. The low-protease assay represents an alternative standardised protocol mimicking conditions found in the gastrointestinal tract. For the high-protease assay, both proteins were incubated with either pepsin or pancreatin and digestion monitored by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and reverse phase-high performance liquid chromatography. The low-protease assay involved gastroduodenal digestion in the presence or absence of phosphatidylcholine (PC). Both beta-casein and beta-Lg were susceptible to hydrolysis by pepsin and pancreatin in the high-protease assay. In contrast, the kinetics of beta-casein digestion in the low-protease assay were slower, beta-Lg being pepsin resistant. During duodenal digestion, beta-Lg was gradually degraded and addition of PC slowed digestion. Subsequently, the reproducibility of the low-protease assay was assessed in 12 independent laboratories by visual assessment of the gels and densitometric analysis: the inter- and intra-laboratory variability was affected by sampling and electrophoresis method employed. The low-protease assay was shown to be reproducible. Future studies will extend these findings using a broader panel of proteins.

The biosynthesis of benzoic acid glucosinolate esters in Arabidopsis thaliana.

The siliques and seeds of Arabidopsis thaliana accumulate a series of glucosinolates containing an alkyl side chain of varying length with a terminal benzoate ester function. The biosynthesis of these unusual nitrogen- and sulfur-containing natural products was investigated by feeding isotopically-labeled precursors to detached flowering stems. Glucosinolates were extracted, purified and analyzed by tandem mass spectrometry. Phenylalanine and benzoic acid were incorporated into the benzoate ester function, and methionine and acetate were incorporated into the aliphatic portion of the side chain in a position-specific manner. The labeling patterns observed were consistent with the chain extension of methionine by a three-step elongation cycle which begins with the condensation of acetyl-CoA with a 2-oxo acid derived from methionine and ends with an oxidative decarboxylation forming a new 2-oxo acid with an additional methylene group. Incorporation of desulfo-4-methylthiobutyl glucosinolate into 4-benzoyloxybutyl olucosinolate suggested chain-extended methionine derivatives are first converted to their corresponding methylthioalkyl glucosinolates with further side chain modification occurring later. Transformation of the methylthiol function to a hydroxyl group is followed by esterification with benzoic acid. The siliques appear to possess the complete machinery for carrying out all of the reactions in the biosyntheis of these complex glucosinolates.

Effect of soldering on metal-porcelain bond strength in repaired porcelain-fused-to-metal castings.

STATEMENT OF PROBLEM: Chemical bonding plays a major role in the adherence between metal and porcelain. The formation of an oxide layer on solder material has not been described in the literature. It is unknown whether the application of solder negatively affects the bond strength between porcelain and metal. PURPOSE: This in vitro study assessed the effect of solder on the bond strength between metal and porcelain. MATERIAL AND METHODS: Forty 20 x 6 x 0.5 mm patterns were divided into test (20) and control (20) groups. Test samples were perforated and repaired with solder, and 2 layers of opaque and dentin porcelain subsequently were applied on all samples. The samples were subjected to a 3-point flexural test on a screw-driven mechanical testing machine at a crosshead speed of 0.5 mm/min. Failure type (adhesive vs cohesive) was quantified by digitizing photographs of test and control samples. Three samples in each group also were examined with an SEM coupled with an x-ray energy-dispersive spectroscopy apparatus (SEM/EDS). Means and standard deviations of loads at failure, sample thickness, and surface area covered with porcelain were calculated, and data were analyzed with Student t test (P < or = 0.05). RESULTS: The mean fracture load for test samples was significantly greater than for control samples (P = 0.0038). Test samples also were significantly thicker (mean thickness difference 0.14 mm) (P = 0.0001). When the data were controlled for thickness by using a multiple linear regression analysis, no significant difference was found (P = 0.68). Test samples had a greater surface area covered with opaque porcelain (P = 0.0006) as determined by visual inspection. CONCLUSION: In this study, soldered and nonsoldered samples did not show any significant difference in porcelain-to-metal bond strength. Visual analysis revealed a significant difference in the amount of porcelain remaining on the fracture surface of the test and control samples; a complete quantitative elemental analysis with SEM/EDS is in progress.

Influence of metal surface finishing on porcelain porosity and beam failure loads at the metal-ceramic interface.

STATEMENT OF THE PROBLEM: Investigators suggest that metals should be finished in 1 direction before porcelain application to minimize metal irregularities and trapped contaminants. These irregularities are thought to be focal points for porosity and crack propagation. PURPOSE: This study investigated the influence of metal finishing and sandblasting on (1) porosity production at the porcelain-metal interface, and (2) porcelain-metal beam failure load. MATERIAL AND METHODS: Eighty cast metal samples were divided into 4 test groups: (A) bidirectional finish/sandblasting; (B) unidirectional finish/sandblasting (C) bidirectional finish only; and (D) unidirectional finish only. The porcelain applied was 1.5 mm thick. Samples were sectioned longitudinally. Half of the samples were subjected to a 3-point flexural test. The remaining samples were sectioned into 4 slices and were examined with a light microscope (x500). Number and diameter of porosities at the metal-porcelain interface were recorded. RESULTS: Mean loads at failure (lbs) were as follows: A, 11.1 +/- 1.3 (5.03 +/- 0.58 Kg); B, 11.2 +/- 1.7 (5. 08 +/- 0.77 Kg); C, 4.0 +/- 1.8 (1.81 +/- 0.81 Kg); and D, 5.0 +/- 2. 1 (2.26 +/- 0.95 Kg). Groups A and B were significantly different from groups C and D (P<.0001). Nonsandblasted samples (C and D) exhibited a separation at the ceramometal interface, which prevented quantification of porosity size and number. Average interface porosity sizes (microm) (A, 8.99 +/- 1.92; B, 10.03 +/- 1.86) showed no significant difference. The mean interface porosity number (A, 62. 3 +/- 16.02; B, 67.4 +/- 10.01) showed no significant difference. CONCLUSION: Direction of metal finishing did not affect the porosity number and size at the ceramometal interface or the beam failure loads. Sandblasting increased the beam failure loads. Nonsandblasted samples showed detachment of the porcelain from the metal.

Biosynthesis of spermidine, a direct precursor of pyrrolizidine alkaloids in root cultures of Senecio vulgaris L.

The polyamine spermidine is an essential biosynthetic precursor of pyrrolizidine alkaloids. It provides its aminobutyl group which is transferred to putrescine yielding homospermidine, the specific building block of the necine base moiety of pyrrolizidine alkaloids. The enzymatic formation of spermidine was studied in relation to the unique role of this polyamine as an alkaloid precursor. S-adenosylmethionine decarboxylase (SAMDC, EC 4.1.1.50) and spermidine synthase (SPDS, EC 2.5.1.16) from root cultures of Senecio vulgaris were partially purified and characterized. The SAMDC-catalyzed reaction showed a pH optimum of 7.5, that of SPDS an optimum of 7.7. The Km value of SAMDC for its substrate S-adenosylmethionine (SAM) was 15 microM, while the apparent Km values of SPDS for its substrates decarboxylated SAM (dSAM) and putrescine were 4 microM and 21 microM, respectively. The relative molecular masses of the two enzymes, determined by gel filtration, were 29000 (SAMDC) and 37000 (SPDS). Studies with various potential inhibitors revealed, for most inhibitors, profiles that were similar to those established with the respective enzymes from other plant sources. However, putrescine which is not known to be an inhibitor of plant SAMDC, strongly inhibited the enzyme from S. vulgaris roots. Spermidine synthase was sensitive to inhibition by its product spermidine. In the presence of the stationary tissue concentrations of the two polyamines (ca. 0.1 mM each) the activities of SAMDC and SPDS would be inhibited by >80%. The results are discussed in relation to the role of spermidine in primary and secondary metabolism of alkaloid-producing S. vulgaris root cultures.

Effect of ring liner and casting ring temperature on the dimension of cast posts.

STATEMENT OF PROBLEM: If a post does not fit passively, binding with the radicular tooth structure occurs, possibly increasing the risk for root fracture. Therefore, it is necessary to fabricate slightly undersized cast posts to allow for passive fit and cement placement. Shrinkage of the mold cavity may be a desired effect during the casting of posts. PURPOSE: This study evaluated the influence of: (1) a cellulose ring liner, and (2) a lower casting temperature of the metal ring, on the dimensions of a cast post. MATERIAL AND METHODS: Eighty plastic posts were divided in 4 equal groups (group 1, 815 degrees C, liner; group 2, 815 degrees C, no liner; group 3, 600 degrees C, liner; and group 4, 600 degrees C, no liner), invested in phosphate-bonded investment and cast with a noble alloy. Posts were measured at 2 locations, the tip and base, before and after casting. RESULTS: Investing in a lined metal ring and using a casting ring temperature of 815 degrees C (group 1) resulted in markedly oversized posts (tip: +25.8 microm; base: +20.3 microm). The absence of the cellulose liner in the casting ring (group 2) decreased the expansion of the investment producing slightly oversized castings (tip: +9.75 microm; base: +4.52 microm). Lined rings that were heated to a final temperature of 600 degrees C (group 3) produced posts that were oversized (tip: +16.65 microm; base: +11.05 microm). Final casting ring temperature of 600 degrees C and the absence of a cellulose liner (group 4) resulted in posts slightly undersized (tip: -4.1 microm; base: -2.2 microm). CONCLUSION: The use of 2 casting ring temperatures (815 degrees C or 600 degrees C) and the influence of a ring liner were investigated. Decreasing the casting ring temperature from 815 degrees C to 600 degrees C and the absence of a ring liner significantly decreased the dimensions of a cast post.

The methionine chain elongation pathway in the biosynthesis of glucosinolates in Eruca sativa (Brassicaceae).

Glucosinolates are nitrogen- and sulfur-containing plant natural products that have become increasingly important in human affairs as flavor precursors, cancer-prevention agents, and crop protectants. While many glucosinolates are biosynthesized from common amino acids, the major glucosinolates in economically important species of the Brassicaceae, such as Brassica napus (oilseed rape), are thought to be formed from chain-elongated derivatives of methionine or phenylalanine. We investigated the chain elongation pathway for methionine that is involved in glucosinolate biosynthesis in Eruca sativa. Isotopically labeled methionine and acetate were administered to cut leaves and the major product, 4-methylthiobutylglucosinolate (isolated as its desulfated derivative), was analyzed by MS and NMR. Administration of ¿U-(13)Cmethionine showed that the entire carbon skeleton of this amino acid, with the exception of the COOH carbon, is incorporated as a unit into 4MTB. Administration of ¿(13)C- and ¿(14)Cacetate gave a labeling pattern consistent with the operation of a three-step chain elongation cycle which begins with the condensation of acetyl-CoA with a 2-oxo acid derived from methionine and ends with an oxidative decarboxylation forming a new 2-oxo acid with one additional methylene group. Administration of ¿(15)Nmethionine provided evidence for the transfer of an amino group to the chain-elongated 2-oxo acid, forming an extended amino acid which serves as a substrate for the remaining steps of glucosinolate biosynthesis. The retention of a high level of (15)N in the products suggests that the amino transfer reactions and the chain elongation cycle are localized in the same subcellular compartment.

Effects of two oral contraceptives on plasma levels of insulin-like growth factor I (IGF-I) and growth hormone (hGH).

UNLABELLED: In 18 healthy women, the effect of two oral contraceptives (OCs) on insulin-like growth factor (IGF-I) and its binding protein-3 (IGFBP-3) and growth hormone (hGH) plasma level were studied before and after intake of either of two OC formulations over 21 days, one containing 2 mg dienogest and 0.03 mg ethinylestradiol (group A) and the other 0.125 mg levonorgestrel and 0.03 mg ethinylestradiol (group B). There was a reduction of the mean IGF-I concentration of 30% (p = 0.008) in the women receiving dienogest-containing pills and 12% (p = 0.006) in women taking the levonogestrel-containing preparation. This difference between drug groups was statistically significant (p = 0.002). A correlation between the control values and the basal-treatment difference (r = 0.945; p = 0.000) was observed only in women of group A. Between basal and treatment cycles the mean plasma levels of hGH remained unchanged in both groups tested, but the 23.5-h integrated mean hGH plasma concentrations (AUC(0-23.5h)) were significantly elevated by 36% (p = 0.016) in comparison to basal values before treatment only in women receiving the levonorgestrel-containing pills. Also, in the women who received the dienogest-containing preparation, the changes of integrated mean plasma level were inversely associated with the control values (r = -0.723; p = 0.025). Neither in group A nor in group B was the mean plasma level of IGFB-3 changed. IN CONCLUSION: the results of the present analysis indicate that hormonal contraceptives can modulate the GH and IGF-I-axis in the reproductive age. Probably the androgenic progestogen levonorgestrel (0.125 mg/day) opposes the estrogen-induced action. In the women who took the dienogest-containing formulations (anti-androgenic progestogen-group A), the extent of individual changes (hGh and IGF-I) depends on the basal level prior to pill intake. Further studies, especially of long-term intake of OCs, are necessary to confirm these results and to assess the practical relevance for possible effects on connective tissue and bone.

Face-bow record without a third point of reference: theoretical considerations and an alternative technique.

Accurate mounting of dental casts is achieved by transferring the tridimensional spatial relationship of the maxillary arch to an articulator. A face-bow is used to transfer this relationship to the articulator, usually by relating the face-bow to a plane of reference. The most common reference plane is the Frankfort plane, which has been assumed to be horizontal when the patient is in the natural head position. The axis-orbitale plane has also been considered horizontal and used as reference. However, it has been shown that both planes are not horizontal, and mounting a maxillary cast according to these planes can result in an inaccurate mounting. This article describes an alternative procedure for face-bow transfer without a plane of reference, and uses the angular relationship between the occlusal plane and the condylar path to mount the maxillary cast on the articulator. The elimination of a reference plane, to which relate the functional determinants of occlusion, avoids an additional source of error during the mounting procedure.

Alternative procedure for making a metal suprastructure in a milled bar implant-supported overdenture.

The predictability of implant-supported prostheses has been established. Although the original Brånemark design has been successfully used in the mandible, esthetic, speech, and hygiene-related problems have been reported in maxillary fixed prostheses. Implant-overdentures can overcome some of the problems encountered in maxillary fixed prostheses. Milled-bar implant-supported overdentures fabricated by electric discharge machining are characterized by stability similar to a fixed prostheses and are removable for hygiene procedures. However, the procedure is costly and requires highly trained technicians. An alternative procedure to produce an accurately fitting metal suprastructure is presented. This procedure does not require additional technical skills and uses instruments and materials that are readily available and relatively inexpensive. The use of simple and easy to replace attachments allows repairs to be performed in the dental office, thus reducing maintenance cost. The overall result is a prosthesis that incorporates the features of a spark erosion overdenture at a fraction of the cost and that is available to a broader patient population.

Long-term reinforced fixed provisional restorations.

Extensive prosthodontic treatment often requires fabrication of long-term provisional restorations. Numerous materials and techniques have been described for prolonged insertion of interim restorations. This article describes a procedure for fabrication of long-term reinforced heat-processed provisional restorations based on a diagnostic wax-up. Reinforced heat-processed provisional restorations reduced flexure, which minimizes progressive loss of cement and diminished the possibility of recurrent decay. Occlusal stability and vertical dimension were maintained because of greater wear resistance. Occlusion, tooth contours, and pontic design developed in the provisional restoration were duplicated in the definitive restoration. The use of a matrix from a diagnostic wax-up facilitated fabrication of the prosthesis, and made the procedure less time-consuming and more predictable.

Immediate implantation of pure titanium implants into extraction sockets of Macaca fascicularis. Part II: Histologic observations.

The purpose of this study was to evaluate immediate implants with implants placed in ossified extraction (control) sites. Histologic observation of immediate and control implants showed similar levels of bone integration at the light microscopic level using ground-section and decalcified thin-section histologic methods. Both histologic methods provided similar information in relation to determination of hard and soft tissue components measured at the interface. The percentage of bone integration was similar in different anatomic regions of the oral cavity. Bone patterns peripheral to the interface region differed, suggesting that special consideration be given to implants placed in the posterior regions of the maxilla and mandible.

Immediate implantation of pure titanium implants into extraction sockets of Macaca fascicularis. Part I: Clinical and radiographic assessment.

Immediate implants have the advantages of few surgical exposures, short treatment time, and maintenance of alveolar bone height and width. The purpose of this study was to compare immediate implants with conventional implants (implants placed into ossified extraction sites) in adult monkeys. Forty-eight implants were placed and allowed to heal for a 6-month period. Following a 7-month loading period, the monkeys were sacrificed, and implant sections were evaluated histologically. Clinical and radiographic measurements showed few significant differences between immediate and control implants.

Fracture strength of full-contoured ceramic crowns and porcelain-veneered crowns of ceramic copings.

This is vitro study compared the fracture strength of Dicor ceramic crowns with a 1 mm axial wall thickness and a 2 mm occlusal thickness with Dicor copings veneered with Vitadur-N or Dicor-Plus porcelain to create similar contours. Tooth preparations for complete crowns were made on human molars with a 10-degree total convergence and a 1 mm shoulder. The artificial crowns were internally etched, given an application of silane, and cemented with a dual-cured resinous cement. After thermocycling, each sample was loaded to failure with a vertical load on the occlusal surface. There were no significant differences in fracture strength between full-contour Dicor crowns and crowns of copings veneered with either Vitadur-N or Dicor-Plus porcelain.

Considerations for planning for and fabricating complete dentures.

During the past year, studies have been completed to advance our knowledge of the diagnosis and treatment of the edentulous patient. The need for pretreatment radiographs on every edentulous patient is not always indicated. Advances in preprosthetic surgery help to improve the predictability of these procedures. New techniques for establishing the vertical dimension of occlusion may allow this to become a more objective measurement. Evaluation of dental materials for use with complete dentures continues, with soft liners and the use of microwave energy for curing denture resin just two areas of interest. A recent study showed a very high prevalence of signs and symptoms of craniomandibular disorders among complete denture wearers. Methods for its treatment are discussed. The use of increased infection control procedures in the dental office and laboratory have created a need to evaluate the effects of disinfecting agents on the dental prosthesis. The removable complete denture remains an effective alternative for the replacement of missing teeth.

Control of microbial contamination with commercially available cleaning solutions.

The elimination of cross-contamination from operatory to laboratory is required for effective infection control. This study determined whether microbial cross-contamination occurs during cleaning of dental prostheses with an ultrasonicator and examined ways to reduce or eliminate the contamination that might occur. The antimicrobial activities of a temporary cement remover and tartar and stain remover were compared with deionized water and a known strong antimicrobial agent. All solutions were assessed without and with ultrasonication. The microbicidal strains were cultured in brain-heart infusion broth, with and without acrylic resin slabs, and the organisms were killed either without or with sonication and without or with acrylic resin slabs. Further testing with natural plaque-contaminated denture materials is warranted.

Oral yeasts, mucosal health, and drug use in an elderly denture-wearing population.

Drug use among the elderly may be a factor in oral mucosal pathology and the prevalence of oral yeasts. This study was designed to evaluate the relationship between oral yeast, denture-bearing mucosal health and drug use, especially drugs with known hyposalivatory side effects. There was an inverse relationship between denture-bearing mucosal health, and hyposalivatory drug use r = -0.41 P less than 0.04. Use of drugs with a hyposalivatory side effect was related to increased numbers of Candida glabrata but not Candida albicans in saliva and at denture stomatitis and denture sore sites.

Dimensional accuracy of denture resin cured by microwave energy.

Microwave processing has several time-saving advantages over conventional processing of denture base resins. Little is known about the adaptation of bases processed by microwave radiation to the cast and the mouth. Comparisons were made in five regions of the palate and ridge. The microwave-processed denture bases had equal or better dimensional accuracy than conventionally processed bases.