Investigation of active matrix- metaloproteinase-8 (aMMP-8) as a reference parameter for path control in antimicrobial photothermal therapy (aPTT) using a split-mouth design.

OBJECTIVES: This retrospective data-collection study aims to explain how the active matrix-metalloproteinase-8-titer (aMMP-titer) influences the immune response of the subject. This is done through monotherapy scaling and root planing (SRP) which is then compared to SRP combined with antimicrobial photothermal therapy (aPTT, Emundo®). METHODS: Data collection was monocentric, randomized and split-mouth based. A study group of twenty patients with chronic periodontal disease with a periodontal pocket depth (PPD) 4 mm ≤ PPD ≤8 mm, a periodontal screening index (PSI: > 3), and a gingival recession ≤2 mm were selected.A diode laser, manufactured by A.R.C. Laser GmbH, with 810 nm wavelength was used. This device implemented three different light transmission systems for transgingival and intra-gingival irradiation. Power settings between 200 and 300 mW were deployed for 10 s during all treatment steps. The photothermic dye of EmunDo® system (A.R.C. Laser GmbH) was infracyaningreen.The adjuvant effect of the antimicrobial photothermal therapy (aPTT) with EmunDo® in combination with conventional SRP on the teeth 15 and 35 was compared with the results of monotherapy SRP on teeth 25 and 45. RESULTS: A reduction of the aMMP-8-titer in gingival crevicular fluid (GCF) was observed in both groups (follow up group and control group) after one month. However; the decrease in the follow up group under SRP in combination with aPTT was significantly more pronounced. The periodontal pocket depths was reduced in both treatment groups. The periodontal probing depth (in mm) shows a larger decrease of the periodontal pocket depth within the follow up group (SPR with aPTT) compared with the control group (SRP). CONCLUSION: The aMMP-8-titer showed differences in both groups prior to and after treatment. Active matrix-metalloproteinase-8 (aMMP-8) as a reference parameter for path control in antimicrobial photothermal therapy (aPTT) seems acceptable.

Comparison of two interdental cleaning aids: A randomized clinical trial.

AIM: The aim of this study was a comparison of the handling and acceptance of two kinds of interdental brushes (interdental brush with a handle [HB] and a newly invented interdental cleaning device [NB]). METHODS AND MATERIALS: In a randomized crossover trial, 40 test subjects with an average number of 23.5 interdental areas were examined. At two appointments with a "washout" period of one week, the volunteers tried out both cleaning tools (HB and NB). They were asked to clean as many interdental spaces as possible. The percentage of spaces, which could be reached, is the IRI (Interdental Reachability Index). Furthermore, subjective impressions were determined. RESULTS: The average IRI using HB was 64% compared to 80% using of NB (P<.001); 62.5% of the volunteers found the cleaning with HB painful and 15% the cleaning with NB. The subjective feeling after cleaning was 1.75 with NB compared to 2.2 with HB (P=.015), grading the feeling, from 1 for "very good" to 5 for "poor." The acceptance of regular interdental hygiene was rated 1.95 in the case of the NB and 2.85 in the case of the HB. After both appointments, the test subjects declared that they would use the NB 3.05 times and the HB 1.78 times a week. CONCLUSION: This study shows that the test subjects reached more interdental spaces with NB than with the HB. NB can improve the usage of interdental brushes. Regarding subjective comfort, participants also favoured NB over HB.

Response of MG63 osteoblasts on bacterial challenge is dependent on the state of differentiation.

The present in vitro study examines molecular processes that are relevant during bone homeostasis after Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis infection with a focus on the differentiation level of osteoblasts. Regenerative processes are often hindered by the recurrence of bacterial infections, which can ultimately provoke a severe destruction of bone tissue. To obtain more detailed insights into such a complex scenario, we have used undifferentiated MG63 osteoblast-like cells as an experimental paradigm to examine the impact of two oral pathogens, A. actinomycetemcomitans and P. gingivalis, on proliferation, cytotoxicity and osteogenic differentiation. Cell culture experiments were performed to analyze cellular behavior. The level of genes interfering with bone tissue integrity (matrix metalloproteinases and their tissue inhibitors) and osteogenic markers (alkaline phosphatase, Runx2, human ß-defensin-2) was compared in undifferentiated versus differentiated MG63 cells using real-time polymerase chain reaction. Functional activity of matrix metalloproteinases was quantified by zymography. Western blot analysis was used to verify the phosphorylation state of mitogen-activated protein kinases p38 and extracellular-signal-regulated kinases 1/2. When co-cultured with undifferentiated MG63 cells, oral pathogens provoked distinct cellular effects. Only A. actinomycetemcomitans reduced cell proliferation, increased cell death, and induced osteogenic differentiation. A comparison of matrix metalloproteinase network stability in the presence of oral pathogens revealed a partial sensitivity towards P. gingivalis but not A. actinomycetemcomitans. So, beside the proof of concept that MG63 cells co-cultured with oral pathogens can serve as an in vitro model for mimicking destructive and regenerative events after bacterial infections, our data indicate that double infections might counterbalance otherwise positive effects.

Oral pathogens change proliferation properties of oral tumor cells by affecting gene expression of human defensins.

The impact of oral pathogens onto the generation and variability of oral tumors has only recently been investigated. To get further insights, oral cancer cells were treated with pathogens and additionally, as a result of this bacterial cellular infection, with human defensins, which are as anti-microbial peptide members of the innate immune system. After cell stimulation, proliferation behavior, expression analysis of oncogenic relevant defensin genes, and effects on EGFR signaling were investigated. The expression of oncogenic relevant anti-microbial peptides was analyzed with real-time PCR and immunohistochemistry. Cell culture experiments were performed to examine cellular impacts caused by stimulation, i.e., altered gene expression, proliferation rate, and EGF receptor-dependent signaling. Incubation of oral tumor cells with an oral pathogen (Porphyromonas gingivalis) and human α-defensins led to an increase in cell proliferation. In contrast, another oral bacterium used, Aggregatibacter actinomycetemcomitans, enhanced cell death. The bacteria and anti-microbial peptides exhibited diverse effects on the transcript levels of oncogenic relevant defensin genes and epidermal growth factor receptor signaling. These two oral pathogens exhibited opposite primary effects on the proliferation behavior of oral tumor cells. Nevertheless, both microbe species led to similar secondary impacts on the proliferation rate by modifying expression levels of oncogenic relevant α-defensin genes. In this respect, oral pathogens exerted multiplying effects on tumor cell proliferation. Additionally, human defensins were shown to differently influence epidermal growth factor receptor signaling, supporting the hypothesis that these anti-microbial peptides serve as ligands of EGFR, thus modifying the proliferation behavior of oral tumor cells.

Diagnostic accuracy of CBCT for periodontal lesions.

OBJECTIVES: The present study compares the diagnostic value of periodontal bone defect images using conventional two-dimensional single-tooth radiographs and three-dimensional cone beam computed tomography (CBCT) images. MATERIALS AND METHODS: Classified periodontal bone defects were prepared on pig mandibles and presented radiographically. Fifteen dentists were instructed to make a diagnosis based on these x-rays, regarding the type and the extent of the bone defects. Subsequently, the results were evaluated and compared to the morphology of the surgically prepared defects as the gold standard. RESULTS: On average, the diagnosis of infrabony defects were 21 %, dehiscence 25 %, and fenestration 33 % more accurate using the three-dimensional projection than with the single-tooth radiograph. Furthermore, the CBCT allows grade II furcation to be captured more accurately. CONCLUSIONS: The results of this study indicate that a considerably more precise analysis of periodontal defects is possible due to the third dimension. Particularly, in the oro-vestibular orientation, defects could be detected significantly more accurate. CLINICAL RELEVANCE: CBCT images offer an advantageous alternative to the conventional single-tooth radiograph while taking the higher exposure of radiation into account.

3D analyses of interface voids in root canals filled with different sealer materials in combination with warm gutta-percha technique.

OBJECTIVES: The aim of the present study was to analyze the formation of voids and gaps in root canals obturated with different sealer materials in combination with warm gutta-percha vertical compaction technique by using BeeFill® 2in1. MATERIALS AND METHODS: Twenty-four single-rooted teeth were collected, and root canals were prepared by using rotary files. All teeth were randomly allocated into three groups. Each group was obturated by using the BeeFill® 2in1 system in combination with Sealapex (non-eugenol, calcium hydroxide polymeric root canal sealer; Kerr Sybron, USA), RoekoSeal (polydimethylsiloxane-based sealer; Roeko, Germany), or 2Seal (epoxy-amine resin-based sealer; VDW, Germany). Following preparation, all teeth were scanned with a micro-computed tomography (CT) scanner, and a three-dimensional reconstruction of the obturated root canals was performed to analyze the volume of interface voids and gaps in the obturated teeth. RESULTS: Statistical analysis demonstrated that the silicon-based sealer RoekoSeal induced significantly less voids and gaps than other tested materials. The amount of voids and gaps significantly was higher in the apical region. CONCLUSIONS: These data indicate that none of the root canal-filled teeth were free of gaps. Teeth obturated with RoekoSeal demonstrated to have the highest quality in terms of voids and gaps formation in combination with the BeeFill® 2in1 obturation system. CLINICAL RELEVANCE: These findings point to the potential benefit of micro-CT analyses for in vitro evaluation of root canal obturation systems and provide further information about sealer materials used in combination with a warm gutta-percha vertical compaction technique.

Defective complex I assembly due to C20orf7 mutations as a new cause of Leigh syndrome.

BACKGROUND: Leigh syndrome is an early onset, progressive, neurodegenerative disorder with developmental and motor skills regression. Characteristic magnetic resonance imaging abnormalities consist of focal bilateral lesions in the basal ganglia and/or the brainstem. The main cause is a deficiency in oxidative phosphorylation due to mutations in an mtDNA or nuclear oxidative phosphorylation gene. METHODS AND RESULTS: A consanguineous Moroccan family with Leigh syndrome comprise 11 children, three of which are affected. Marker analysis revealed a homozygous region of 11.5 Mb on chromosome 20, containing 111 genes. Eight possible mitochondrial candidate genes were sequenced. Patients were homozygous for an unclassified variant (p.P193L) in the cardiolipin synthase gene (CRLS1). As this variant was present in 20% of a Moroccan control population and enzyme activity was only reduced to 50%, this could not explain the rare clinical phenotype in our family. Patients were also homozygous for an amino acid substitution (p.L159F) in C20orf7, a new complex I assembly factor. Parents were heterozygous and unaffected sibs heterozygous or homozygous wild type. The mutation affects the predicted S-adenosylmethionine (SAM) dependent methyltransferase domain of C20orf7, possibly involved in methylation of NDUFB3 during the assembly process. Blue native gel electrophoresis showed an altered complex I assembly with only 30-40% of mature complex I present in patients and 70-90% in carriers. CONCLUSIONS: A new cause of Leigh syndrome can be a defect in early complex I assembly due to C20orf7 mutations.

Penetration depth of a dye marker into dentine using a novel hydrodynamic system (RinsEndo).

AIM: To investigate the efficiency of a hydrodynamic irrigation system compared with conventional cleansing techniques in root canals. METHODOLOGY: Forty-five freshly extracted single-rooted teeth were de-coronated and their root canals were enlarged to size 30 at the apex. The teeth were randomly divided into three groups (n = 15) for the final rinsing sequence using 2% NaOCl plus acid fuchsin: group I: static application of irrigant, 3 min; group II: flushing with a syringe; 5-mL NaOCl, 1 min; group III: RinsEndo-system; 5-mL NaOCl, 50 s. Apical extrusion was documented photographically. The roots were sectioned at 2, 4, 6 and 8 mm from their apices and the penetration depths of dye into dentine measured, using a stereomicroscope. Wilcoxon's test and Pearson's chi-squared test were employed to prove statistic relevance. RESULTS: Greater dye penetration depth into the dentinal tubules was achieved when employing hydrodynamic rinsing procedures. Using this technique, 23% of the specimens were penetrated for more than 50% of their dentine thickness, whereas the results for flushing with a syringe were 12% (static application, 7%). No penetration of dentine occurred in 63% of specimens with static application, 39% flushing with a syringe and 15% using the hydrodynamic system (P < 0.05 Pearson's chi-squared test). Apical extrusion occurred more frequently after hydrodynamic rinsing (extruded specimens: RinsEndo = 80%; static application/flushing with a syringe = 13%; P < 0.05 Pearson's chi-squared test). CONCLUSIONS: Hydrodynamic rinsing demonstrated an improvement over conventional methods in terms of dentine penetration of a dye marker. A higher risk of apical extrusion with the RinsEndo-system was evident.

Clinical and microbiological effects of topical subgingival application of hyaluronic acid gel adjunctive to scaling and root planing in the treatment of chronic periodontitis.

BACKGROUND: Hyaluronic acid (HA) has shown anti-inflammatory effects in gingivitis therapy. The potential benefits of local subgingival application of HA adjunctive to scaling and root planing (SRP) were evaluated in this study. METHODS: Twenty patients with chronic periodontitis were included in this split-mouth study. Sulcus fluid flow rate (SFFR) and sulcus bleeding index were monitored at baseline and after 1, 2, 3, 4, 5, 6, and 12 weeks; probing depth and clinical attachment level were monitored at baseline and 6 and 12 weeks. Subgingival plaque samples were also taken at these same three appointments to determine the presence of Actinobacillus actinomycetemcomitans, Porphyromonas gingivalis, Prevotella intermedia, Tannerella forsythensis, and Treponema denticola. All patients were treated with full-mouth scaling and root planing (SRP); in addition, an HA gel was administered subgingivally in the test sites every week for 6 weeks. RESULTS: An improvement of all clinical variables was observed (P < 0.05) in both groups. Clinically, no difference between test and control sites could be found. No difference between test and control sites was seen in the tested microorganisms. CONCLUSIONS: No clinical or microbiological improvement was achieved by the adjunctive use of HA gel compared to SRP alone. Only SFFR was affected by the use of HA in terms of a more rapid reduction of SFFR in the test sites.

Effect of diode laser and fluoride varnish on initial surface demineralization of primary dentition enamel: an in vitro study.

INTRODUCTION: Previous investigations have demonstrated improved enamel caries resistance after laser irradiation. The purpose of this in vitro study was to assess the caries-preventive potential of 809 nm diode laser treatment of the enamel of primary teeth compared to topical fluoride application. METHODS: Eighty samples of sound primary teeth were embedded in plastic and stored in saline solution. The enamel surface of 80 samples was polished in an area of 2 x 2 mm. These tooth specimens were randomly assigned to one control and three test groups: (1) no treatment/control; (2) application of 0.1 mg of fluoride varnish (Duraphat) for 6 h; (3) diode laser application (809 nm, 140 mJ, 50 Hz, Ø 600 micro m fibre, contact mode, absorber, 1 min; ORA-LASER01 I.S.T.); and (4) combined application - laser/fluoride varnish. Caries-like lesions were created by pH-cycling. After lesion formation, longitudinal sections were taken and examined by polarized light microscopy. RESULTS: In the control group, all samples showed lesions up to 30 micro m in depth. After laser application, lesions could be identified in 15 out of 20 samples. Topical fluoride treatment in groups 2 (varnish) and 4 (laser/varnish) completely inhibited the development of caries-like lesions in all samples. CONCLUSION: In this in vitro investigation, topical fluoride treatment enhances the resistance of sound enamel of primary teeth more effectively than diode laser application.

[Clinical study of hyaluronic acid in the treatment of chronic periodontitis].

OBJECTIVE: Clinical effects in local application of hyaluronic acid (HA) adjunctive to scaling and root planing (SRP) were evaluated. METHODS: In this cross over design study 20 patients with chronic periodontitis were included. Plaque index, sulcus-fluid-flow-rate (SFFR), sulcus bleeding index, probing pocket depth and attachment level were monitored. All patients were treated with full mouth SRP, in addition a HA gel was administered subgingivally in the test site every week for 6 weeks. RESULTS: A statistically significant improvement of all clinical test parameters was observed in both groups (P < 0.05). Clinically, no significant difference between test and control group could be found. However, the SFFR decreased significantly faster in the test group. CONCLUSION: No post-inflammatory tissue regeneration could be achieved by the adjunctive use of HA gel to SRP in the patients with chronic periodontitis. Due to SFFR a control of local inflammation can be achieved quickly.

Osteotomy with 80-micros CO2 laser pulses--histological results.

Haemostatic and aseptic effects and intricate cut geometry are beneficial aspects of non-contact laser osteotomy. Collateral thermal damage, however, has severely limited the use of conventional lasers. The purpose of this study was to test the side effects on bone after cutting it with short CO2 laser pulses and simultaneous application of a fine air-water spray. The 10.6 microm CO2 laser emitted 80 micros pulses of 46 mJ energy, f=100 Hz, focused to a spot diameter of 130 ìm. Scan rate amounted to 40 mm/s. To approximate live conditions 10 samples of cortical bone and 10 rib segments were prepared immediately after sacrificing of pigs. A reference cut with a bandsaw and three laser cuts with an increasing number of beam passes (4, 16, 64) were performed on each sample. Half of the samples were decalcified in EDTA. The others were embedded in plastic to cut non-decalcified sections. The laser incisions were not accompanied by carbonisation. The incisions with slightly convergent walls were 150 ìm wide. The depths of the cavities increased with the number of the beam passes from approximately 0.5 mm (4 passes) to 3 mm (64 passes). At the border of the incisions two narrow zones of damage were noted: an amorphous intensively stained zone of 1-3 microm width and a wider, also sharply demarcated but faintly stained zone of 7-10 microm. A broader zone of about 50 microm was characterised by empty lacunae and osteocyte damage. These effects were not predictable; intact osteocytes were also observed near to the cut surface. Polarised light microscopy showed no alterations in the inorganic structure of the bone at the cut borders. The histological results indicated only minimal damage to bone ablated at the specified parameters. The described laser procedure might have advantages over mechanical instruments.

Endodontic and surgical treatment of a geminated maxillary incisor.

AIM: A geminated maxillary incisor required complex multidisciplinary treatment to preserve health and restore aesthetics. This report describes the coordination of coronal division, root extraction, bone grafting, endodontic treatment and orthodontics in achieving treatment goals. It is the intention of this report to show how a difficult case could be managed by properly coordinated multidisciplinary care. SUMMARY: Pulp testing of the geminated central right maxillary incisor gave a normal response. Radiographic investigation indicated a connection of the pulp chambers. Both the mesial and distal root were filled with gutta-percha. The connection between the two root canals was sealed with a flowable dentine-bonded resin. After dividing the crown with a diamond bur, the mesial part of the tooth was removed and the extraction socket was filled with beta-tricalcium phosphate ceramic (Cerasorb). Radiographs taken immediately after surgery and after 6 months showed no periodontal or periapical lesions. No signs of external resorption were identified. The diastema between the central incisors was closed by orthodontic treatment. KEY LEARNING POINTS: Geminated teeth may present aesthetic and functional problems which require multidisciplinary care. Careful clinical and radiographic examination is essential to decide on the fate of the coronal and root halves involved. A proper coordination between endodontic and surgical treatment may result in maintaining one tooth half, even if a midroot connection between the pulp chambers becomes evident. Orthograde endodontic treatment, hemisection and orthodontics may solve the aesthetic problem of a geminated tooth.

The possibility of detecting subgingival calculus by laser-fluorescence in vitro.

The aim of the present study was to investigate laser-fluorescence effects on the root surface. The possibility of detecting subgingival calculus should be evaluated to control complete calculus removal after scaling and root planing. Twenty freshly extracted human teeth, partially covered with calculus on the root surface, were fixed on a translation stage. The specimens were irradiated by light from a laser source (diode laser, <1 mW) with a wavelength of 655 nm using the Diagnodent trade mark -system. To simulate the conditions in a periodontal pocket during the measurement procedure, the teeth were covered by a layer of physiological saline solution or blood in order to determine the influence of different inflammatory fluids. The root surface was scanned on a line of about 1 cm in steps of 0.5 mm. The start and end was marked by a notch. The results of the laser-fluorescence detection were compared to clinical and histological findings. Clinically apparent calculus on the root surface was always accompanied with a statistically significant increase of laser-fluorescence values ( p < 0.05). These values were not influenced by the different fluids ( p < 0.05). The laser-fluorescence values were reproducible ( p > 0.05). After scaling, when no debris remained at the root surface, laser-fluorescence values decreased significantly ( p < 0.05). The histological findings supported the clinical results. In conclusion, the present study indicates that laser-fluorescence values on the root surface are strongly correlated to the presence of calculus. This fluorescence effect can basically be used to develop a new detecting system for subgingival calculus in periodontal pockets.

Clinical and microbiological effects of local chlorhexidine applications.

AIM: To study the effects of an application of chlorhexidine varnish (40%) on dental plaque. DESIGN: Randomised controlled clinical trial. SUBJECTS: 40 patients (50% females), aged 25-34 years after undergoing any necessary restorative treatment. Exactly one half of these individuals practiced good oral hygiene, the efforts of the others were poor. METHOD: Test groups received a chlorhexidine varnish application, the control group received a placebo varnish. Initially and after 2 and 6 weeks, a modified Dentocult SM-test, bleeding on probing, and a plaque index (Quigley-Hein) were recorded. RESULTS: In contrast to the control group, improvements in plaque index and bleeding on probing scores were found in patients with poor oral hygiene. The results of the Dentocult SM-tests showed a considerable reduction of streptococcus colonisation. The test group with good oral hygiene showed only slight improvement of the examined indices after chlorhexidine application. The index parameters of all patients of the placebo groups remained unchanged over the examination time period. CONCLUSION: The application of a chlorhexidine varnish reduced the quantity of the Streptococcus mutans colonies significantly and improved clinical parameters in patients with elevated plaque accumulation.

Cavity preparation using a superpulsed 9.6-microm CO2 laser--a histological investigation.

BACKGROUND AND OBJECTIVES: The superpulsed 9.6-microm CO(2) laser is an effective laser for ablating dental tissues and decay. This histological study compares laser class V preparations with conventional treatment to evaluate the resulting formation at the cavity walls. STUDY DESIGN/MATERIALS AND METHODS: Four class V preparations (one made with a diamond drill and three with the CO(2) laser (9.6 microm, 60 microseconds pulse width, 40 mJ pulse energy, 100 Hz, integrated scanner system, water cooling) were performed on ten extracted teeth. The cavities were filled with a composite resin partly including enamel and dentine conditioning. RESULTS: After laser preparation, no cracks or signs of carbonisation were detected. The results were comparable to those attained with conventional treatment. Following cavity filling without prior conditioning, gaps were noted at the cavosurface indicating a lack of adhesion. Dentinal bonding decreased gap formation significantly. CONCLUSION: The 9.6-microm CO(2) laser is an effective tool for cavity preparation.

Er:YAG laser scaling of diseased root surfaces.

BACKGROUND: The removal of calculus and plaque is an essential component of a therapeutic approach to control periodontal disease. Er:YAG laser scaling was recently introduced as an alternative to conventional scaling procedures. In this histological study, the effects of laser instrumentation of diseased root surfaces are compared to mechanical removal of plaque and calculus with ultrasonic instruments and scalers. METHODS: Areas of subgingival calculus were identified on 40 freshly extracted human teeth. Each of these areas was randomly divided into 2 equal parts. The control site was treated either with scaling and root planing or with an ultrasonic instrument. The test site was cleaned using an Er:YAG laser according to the manufacturer's instructions. The end point of debridement was the inability to mechanically or visibly detect any remaining calculus. After pre- and postsurgical photographs and impressions for scanning electronic microscopic investigation, a plastic embedding technique was used to cut the undecalcified roots into 15 microm thick sections. RESULTS: Clinically and histologically, scaling resulted in complete debridement at all samples, producing a smooth root surface. At the test sites, laser scaling was accompanied by an increased removal of tissue and roughened surfaces. CONCLUSIONS: Laser scaling results in an increased loss of cementum and dentin, which should be taken into account in clinical situations.

Phosphatidylglycerophosphate synthases from Arabidopsis thaliana.

Two Arabidopsis thaliana genes were shown to encode phosphatidylglycerophosphate synthases (PGPS) of 25.4 and 32.2 kDa, respectively. Apart from their N-terminal regions, the two proteins exhibit high sequence similarity. Functional expression studies in yeast provided evidence that the 25.4 kDa protein is a microsomal PGPS while the 32.2 kDa protein represents a preprotein which can be imported into yeast mitochondria and processed to a mature PGPS. The two isozymes were solubilized and purified as fusion proteins carrying a His tag at their C-terminus. Enzyme assays with both membrane fractions and purified enzyme fractions revealed that the two A. thaliana isozymes have similar properties but differ in their CDP-diacylglycerol species specificity.

Functional characterization of beta-ketoacyl-CoA synthase genes from Brassica napus L.

Seed-specifically expressed beta-ketoacyl-CoA synthase genes of Brassica napus (Bn-FAE1.1 genes) were cloned from two cultivars, namely Askari, a high-erucic-acid type, and Drakkar, a low-erucic-acid type. The genes from the two cultivars were found to be nearly identical. They encode proteins of 507 amino acids, the sequences of which differ only at position 282. The Bn-FAE1.1 gene of Askari, unlike that of Drakkar, was functionally expressed in yeast cells suggesting that the single amino acid exchange effects the low erucic acid phenotype at the E1 gene locus. In yeast cells the beta-ketoacyl-CoA synthase of Askari elongated not only oleoyl but also palmitoleoyl groups as well as saturated acyl groups in such a way that monounsaturated acyl groups of 22 carbons and saturated ones of 26 carbons were formed as main products. A reporter gene fused to the promoter region of the Bn-FAE1.1 gene from Askari showed seed-specific expression in transgenic rapeseed plants. Over-expression of the coding region of the Askari gene in developing seeds of transgenic Drakkar plants resulted in a significant increase in the levels of eicosenoic acid and erucic acid esterified in the seed oil. On the other hand, in transgenic high-erucic-acid rapeseed plants the increase in erucic acid level was at most 60% although the chimeric Bn-FAE1.1 gene was co-expressed with an erucoyl-CoA-specific lysophosphatidate acyltransferase gene enabling trierucoyl glycerol to accumulate in the seed oil.

In vitro analysis of the effects of acid or laser etching on microleakage around composite resin restorations.

OBJECTIVES: The aim of the present study was to determine the effect of two conditioning methods, phosphoric acid and XeCl laser application on microleakage at composite resin restorations. METHODS: Class II MOD (mesio-occluso-distal) cavities were prepared on 90 extracted human wisdom teeth with one proximogingival margin on enamel and the other on dentin. Phosphoric acid (37%) and a XeCl excimer laser system were used for the etching procedure. The prepared teeth were randomly assigned to six groups: In group 1, acid-etching of the enamel margins was followed by application of enamel bonding, in group 2 additionally a dentin bonding system was used. In group 3, acid-etching of the enamel and dentin (total etch) was followed by the application of dentin bonding. In group 4, laser-etching of enamel margins was followed by the application of enamel bonding, in group 5 dentin bonding was used additionally. In group 6, laser-etching of enamel and dentin margins was followed by the application of dentin bonding. All restorations were placed incrementally and polymerized with a halogen light source. After restoration, all samples were thermocycled and stained in 0.5% basic fuchsin and sectioned longitudinally in a mesio-distal plane. Microleakage was scored by means of a stereomicroscope. The margins of the restorations at tooth-resin interface were examined with a scanning electron microscope. RESULTS: In all groups, marginal leakage was more extensive at the gingival margin that was in dentin than at the gingival margin in enamel. Microleakage scores and the gap at the tooth-resin interface were significantly lower in all acid-etched groups than in laser-etched groups. CONCLUSION: Acid-etch technique is more effective than the investigated laser-etch technique at 308nm concerning the reduction of marginal leakage in composite restorations.