Soluble ST2 regulation by rhinovirus and 25(OH)-vitamin D3 in the blood of asthmatic children.

Paediatric asthma exacerbations are often caused by rhinovirus (RV). Moreover, 25(OH)-vitamin D3 (VitD3) deficiency during infancy was found associated with asthma. Here, we investigated the innate immune responses to RV and their possible modulation by 25(OH)-VitD3 serum levels in a preschool cohort of children with and without asthma. The innate lymphoid cell type 2 (ILC2)-associated marker, ST2, was found up-regulated in the blood cells of asthmatic children with low serum levels of 25(OH)-VitD3 in the absence of RV in their airways. Furthermore, in blood cells from control and asthmatic children with RV in their airways, soluble (s) ST2 (sST2) protein was found reduced. Asthmatic children with low 25(OH)-VitD3 in serum and with RV in vivo in their airways at the time of the analysis had the lowest sST2 protein levels in the peripheral blood compared to control children without RV and high levels of 25(OH)-VitD3. Amphiregulin (AREG), another ILC2-associated marker, was found induced in the control children with RV in their airways and low serum levels of 25(OH)-VitD3. In conclusion, the anti-inflammatory soluble form of ST2, also known as sST2, in serum correlated directly with interleukin (IL)-33 in the airways of asthmatic children. Furthermore, RV colonization in the airways and low serum levels of 25(OH)-VitD3 were found to be associated with down-regulation of sST2 in serum in paediatric asthma. These data indicate a counter-regulatory role of 25(OH)-VitD3 on RV-induced down-regulation of serum sST2 in paediatric asthma, which is relevant for the therapy of this disease.

Development of a DNA Macroarray for the Detection and Identification of Fungal Pathogens Causing Decline of Young Grapevines.

Young vine decline (YVD) is a complex disease caused by at least 51 different fungi and responsible for important economic losses to the grapevine industry worldwide. YVD fungi are known to occur in planting material. Hence, detection prior to planting is critical to assure longevity of newly established vineyards. A DNA macroarray based on reverse dot-blot hybridization containing 102 oligonucleotides complementary to portions of the ß-tubulin region was developed for detection of YVD fungi. Specificity of the array was first evaluated against 138 pure fungal cultures representing 72 different taxa from nine genera, including 37 YVD species. In total, 61 species, including 34 YVD pathogens, were detected and identified by the array. The detection limit of the array was below 0.1 pg of genomic DNA. The array was validated against artificially inoculated canes and soil and commercial planting material, with the latter showing a high incidence of YVD fungi in nursery plants otherwise not detected by traditional plating and culturing. This DNA array proved to be a rapid and specific tool to simultaneously detect and identify most YVD fungi in a single test, which has the potential to be used in commercial diagnostics or by the grapevine nursery industry to determine the health status of the planting material.

Harnessing the lysosome-dependent antitumor activity of phenothiazines in human small cell lung cancer.

Phenothiazines are a family of heterocyclic compounds whose clinical utility includes treatment of psychiatric disorders as well as chemotherapy-induced emesis. Various studies have demonstrated that these compounds possess cytotoxic activities in tumor cell lines of different origin. However, there is considerable confusion regarding the molecular basis of phenothiazine-induced cell death. Lung cancer (LC) remains one of the most prevalent and deadly malignancies worldwide despite considerable efforts in the development of treatment strategies, especially new targeted therapies. In this work, we evaluated the potential utility of phenothiazines in human LC. We show that phenothiazines as single treatment decreased cell viability and induced cell death preferentially in small cell lung carcinoma (SCLC) over non small cell lung carcinoma (NSCLC) cell lines. Sensitivity to phenothiazines was not correlated with induction of apoptosis but due to phenothiazine-induced lysosomal dysfunction. Interestingly, the higher susceptibility of SCLC cells to phenothiazine-induced cell death correlated with an intrinsically lower buffer capacity in response to disruption of lysosomal homeostasis. Importantly, this effect in SCLC occurred despite mutation in p53 and was not influenced by intrinsic sensitivity/resistance toward conventional chemotherapeutic agents. Our data thus uncovered a novel context-dependent activity of phenothiazines in SCLC and suggest that phenothiazines could be considered as a treatment regimen of this disease, however, extended cell line analyses as well as in vivo studies are needed to make such conclusion.

Grapevine Trunk Diseases in British Columbia: Incidence and Characterization of the Fungal Pathogens Associated with Black Foot Disease of Grapevine.

Black foot disease of grapevines, caused by several fungal species in the genera Campylocarpon, Cylindrocarpon, Cylindrocladiella, and Ilyonectria, causes significant economic losses to the grapevine industry worldwide. This study represents the first attempt to identify and characterize the fungal pathogens associated with black foot disease of grapevines in British Columbia (BC). Field surveys conducted throughout all grape-growing regions in BC that included assessment of foliar symptomatology and isolations from symptomatic vines showed Cylindrocarpon/Ilyonectria spp. occurred in 32 of 90 (35.5%) young vineyards surveyed (≤8 year old) and in 41 of 215 (19%) samples collected. In 20 of the 41 (48.8%) samples, Cylindrocarpon/Ilyonectria spp. were the sole fungi isolated from symptomatic tissue. In the rest of the samples, black foot fungi were found to primarily coexist with fungal taxa associated with Petri disease of grapevines. Colony and conidia phenotypical characterization, along with DNA analyses of the internal transcribed spacer region (ITS1-5.8S-ITS2) of the rDNA, and part of the ß-tubulin and translation elongation factor 1-α genes, revealed five different black foot fungi occurring in declining young vines in BC, namely Cylindrocarpon pauciseptatum, Ilyonectria liriodendri, Ilyonectria macrodidyma, Ilyonectria robusta, and Ilyonectria torresensis. Pathogenicity studies showed all five species to be highly virulent in the grapevine rootstock cultivar 3309C. Overall, I. liriodendri and I. macrodidyma were the most virulent species when inoculated in Vitis vinifera 'Chardonnay' and rootstock 3309C.

Grapevine Trunk Diseases in British Columbia: Incidence and Characterization of the Fungal Pathogens Associated with Esca and Petri Diseases of Grapevine.

Esca and Petri disease are two economically important grapevine diseases worldwide. This study reports for the first time the occurrence of both diseases on grapevines in British Columbia (BC) and subsequently in Canada. Visual assessment of 55,699 vines in 118 vineyards revealed a low incidence of esca with only 104 (0.2%) vines showing foliar symptoms. Young vine decline (YVD) was observed in 1,910 (7.8%) of 24,487 monitored young vines and in 52 (8%) of 654 young vines used as re-plants in mature vineyards. In 8 of 51 monitored young vineyards, YVD-affected vines ranged between 15 and 55%. Morphological studies along with DNA analyses of the ITS1-5.8S-ITS2, and part of the ß-tubulin, actin, and translation elongation factor 1-α gene regions, allowed us to identify Cadophora luteoolivacea, Phaeomoniella chlamydospora, Phaeoacremonium iranianum, Togninia fraxinopennsylvanica, Togninia minima, and the novel species Phaeoacremonium canadense and Phaeoacremonium roseum from esca and Petri disease infected vines in BC. This study includes for the first time the EF1-α DNA marker in Phaeoacremonium spp. delineation. Pathogenicity studies showed all seven fungi to cause vascular symptoms similar to those observed in esca and Petri disease infected vines. Additionally, the "tiger-stripes" foliar symptom of esca was successfully reproduced when healthy potted vines were inoculated with BC isolates of Pa. chlamydospora, Pm. canadense, Pm. iranianum, T. fraxinopennsylvanica, and T. minima.

Resistance to DNA-damaging treatment in non-small cell lung cancer tumor-initiating cells involves reduced DNA-PK/ATM activation and diminished cell cycle arrest.

Increasing evidence suggests that tumor-initiating cells (TICs), also called cancer stem cells, are partly responsible for resistance to DNA-damaging treatment. Here we addressed if such a phenotype may contribute to radio- and cisplatin resistance in non-small cell lung cancer (NSCLC). We showed that four out of eight NSCLC cell lines (H125, A549, H1299 and H23) possess sphere-forming capacity when cultured in stem cell media and three of these display elevated levels of CD133. Indeed, sphere-forming NSCLC cells, hereafter called TICs, showed a reduced apoptotic response and increased survival after irradiation (IR), as compared with the corresponding bulk cell population. Decreased cytotoxicity and apoptotic signaling manifested by diminished poly (ADP-ribose) polymerase (PARP) cleavage and caspase 3 activity was also evident in TICs after cisplatin treatment. Neither radiation nor cisplatin resistance was due to quiescence as H125 TICs proliferated at a rate comparable to bulk cells. However, TICs displayed less pronounced G2 cell cycle arrest and S/G2-phase block after IR and cisplatin, respectively. Additionally, we confirmed a cisplatin-refractory phenotype of H125 TICs in vivo in a mouse xenograft model. We further examined TICs for altered expression or activation of DNA damage repair proteins as a way to explain their increased radio- and/or chemotherapy resistance. Indeed, we found that TICs exhibited increased basal γH2AX (H2A histone family, member X) expression and diminished DNA damage-induced phosphorylation of DNA-dependent protein kinase (DNA-PK), ataxia telangiectasia-mutated (ATM), Krüppel-associated protein 1 (KAP1) and monoubiquitination of Fanconi anemia, complementation group D2 (FANCD2). As a proof of principle, ATM inhibition in bulk cells increased their cisplatin resistance, as demonstrated by reduced PARP cleavage. In conclusion, we show that reduced apoptotic response, altered DNA repair signaling and cell cycle perturbations in NSCLC TICs are possible factors contributing to their therapy resistance, which may be exploited for DNA damage-sensitizing purposes.

Chemosensitization by phenothiazines in human lung cancer cells: impaired resolution of γH2AX and increased oxidative stress elicit apoptosis associated with lysosomal expansion and intense vacuolation.

Chemotherapy resistance poses severe limitations on the efficacy of anti-cancer medications. Recently, the notion of using novel combinations of 'old' drugs for new indications has garnered significant interest. The potential of using phenothiazines as chemosensitizers has been suggested earlier but so far our understanding of their molecular targets remains scant. The current study was designed to better define phenothiazine-sensitive cellular processes in relation to chemosensitivity. We found that phenothiazines shared the ability to delay γH2AX resolution in DNA-damaged human lung cancer cells. Accordingly, cells co-treated with chemotherapy and phenothiazines underwent protracted cell-cycle arrest followed by checkpoint escape that led to abnormal mitoses, secondary arrest and/or a form of apoptosis associated with increased endogenous oxidative stress and intense vacuolation. We provide evidence implicating lysosomal dysfunction as a key component of cell death in phenothiazine co-treated cells, which also exhibited more typical hallmarks of apoptosis including the activation of both caspase-dependent and -independent pathways. Finally, we demonstrated that vacuolation in phenothiazine co-treated cells could be reduced by ROS scavengers or the vacuolar ATPase inhibitor bafilomycin, leading to increased cell viability. Our data highlight the potential benefit of using phenothiazines as chemosensitizers in tumors that acquire molecular alterations rendering them insensitive to caspase-mediated apoptosis.

Bond strength of luting materials to ceramic crowns after different surface treatments.

The aim of present study was to evaluate the effect of different pre-treatments of alumina and zirconia copings on the bond strength of different luting materials. Pull out tests was performed on 60 alumina and 80 zirconia copings. Randomly selected, copings were divided in groups of i) un-treated alumina and zirconia copings, (n=20) ii) alumina and zirconia copings sandblasted with 50 or 110 microm alumina particles respectively (n=20), iii) zirconia copings treated with monolayer of glass pearls fused to the inner surface (n=20), iv) zirconia copings treated with silanized glass pearls (n=10). Zinc phosphate, Panavia 21 and VarioLink II were used for cementation. Pull out tests were done in an Instron universal testing machine with a speed of 0.5 mm/min and fracture loads was measured in N. Untreated zirconia copings cemented with zinc phosphate showed significantly higher bond strength values compared to those with sandblasted surfaces. No difference was seen between untreated alumina copings and those with sandblasted surfaces. Sandblasting decreased bond strength of zirconia and alumina copings. Glass pearls increased bond strength of zirconia copings, which was even better after silanization. Variolink II in combination with alumina gave significantly lower bond strength.

Fracture strength of two oxide ceramic crown systems after cyclic pre-loading and thermocycling.

The aim of the present study was to investigate the fracture resistance of zirconia crowns and to compare the results with crowns made of a material with known clinical performance (alumina) in away that reflects clinical aspects. Sixty crowns were made, 30 identical crowns of alumina and 30 of zirconia. Each group of 30 was randomly divided into three groups of 10 crowns that were to undergo different treatments: (i) water storage only, (ii) pre-loading (10 000 cycles, 30-300 N, 1 Hz), (iii) thermocycling (5-55 degrees , 5000 cycles) + pre-loading (10 000 cycles, 30-300 N, 1 Hz). Subsequently, all 60 crowns were subjected to load until fracture occurred. There were two types of fracture: total fracture and partial fracture. Fracture strengths (N) were: group 1, alumina 905/zirconia 975 (P = 0.38); group 2, alumina 904/zirconia 1108 (P < 0.007) and group 3, alumina 917/zirconia 910 (P > 0.05). Total fractures were more frequent in the alumina group (P < 0.01). Within the limitations of this in vitro study, it can be concluded that there is no difference in fracture strength between crowns made with zirconia cores compared with those made of alumina if they are subjected to load without any cyclic pre-load or thermocycling. There is, however, a significant difference (P = 0.01) in the fracture mode, suggesting that the zirconia core is stronger than the alumina core. Crowns made with zirconia cores have significantly higher fracture strengths after pre-loading.

Shade selection performed by novice dental professionals and colorimeter.

The objective of this study was to test inter-observer variability in shade selection for porcelain restorations, using three different shade guides: Vita Lumin Vacuum, Vita 3D-Master and Procera. Nineteen young dental professionals acted as observers. The results were also compared with those of a digital colorimeter (Shade Eye Ex; Shofu, Japan). Regarding repeatability, no significant differences were found between the three shade guides, although repeatability was relatively low (33-43%). Agreement with the colorimetric results was also low (8-34%). In conclusion, shade selection shows moderate to great inter-observer variation. In teaching and standardizing the shade selection procedure, a digital colorimeter may be a useful educational tool.

The use of generic failure frequencies in QRA: the quality and use of failure frequencies and how to bring them up-to-date.

Quantitative Risk Assessment (QRA) is a method which is often used in the chemical industry and, in some countries, also in land-use planning. In QRA calculations the frequency of an accident scenario is most often assessed by a generic failure frequency approach. The credibility and validity of the failure frequencies used in the Netherlands for land-use planning is evaluated by means of an historical review. Furthermore, the possibility is presented how these generic data can be revised and updated.

Genes differentially expressed in prostate cancer.

Because of the heterogeneity of prostate cancer knowledge about the genes involved in prostate carcinogenesis is still very limited. Previously, the use of novel high-throughput technologies offered the possibility to investigate broad gene expression profiles and thus helped to improve understanding of the molecular basis of prostate disease. Many candidate genes have been identified so far which have a more or less strong effect on prostate cancer. This vast number of gene expression changes show that it is unlikely that only one gene promotes prostate cancer. Conversely, it seems more likely that a broad network of molecular changes is involved in the complex cascade of events which lead to tumour formation and progression, respectively. A few of these novel molecular targets are currently under clinical evaluation. This paper gives an overview of several interesting candidate genes which may be useful as improved biomarkers for diagnosis or as targets for developing novel treatment methods.

First Report of Brown Rot in Wine Grapes Caused by Monilinia fructicola in Canada.

A survey was conducted in 2001 and 2002 to determine incidence of fruit pathogens in wine grapes (Vitis vinifera), an important crop in the southern interior of British Columbia (BC), Canada. Grape clusters were sampled every 2 weeks from June to October at eight vineyard sites located from Osoyoos in the south to Kelowna, approximately 100 km to the north. In the laboratory, the berry clusters were surface disinfested for 0.5 min in 70% ethanol, followed by 1 min in 0.5% sodium hypochlorite, and rinsed twice in sterile distilled water. The berries were placed on potato dextrose agar (PDA) amended with 15 ml/liter of 85% lactic acid and incubated at 20°C for 1 week. During the 2002 survey, a fungus resembling Monilinia fructicola (G. Wint.) Honey was observed sporulating on immature 'Pinot noir' grapes from Kelowna that were sampled on 14 August. Later in the growing season, a similar fungus was detected on 'Riesling' grapes from Summerland sampled on 11 September. There was no evidence of brown rot near the vineyard in Kelowna, but diseased stonefruit were present near the vineyard in Summerland. Subsequent identification of the fungus from 'Riesling' as M. fructicola was based on morphological characters and DNA sequence data for the internal transcribed spacer (ITS) regions of the nuclear ribosomal rRNA genes. The sequenced isolate was deposited in the Canadian Collection of Fungus Cultures as DAOM 231119, and the ITS sequence was accessioned in GenBank as AY289185. Colony growth on PDA was rapid and in concentric rings with the colony margin complete, microconidia abundant, and macroconidia 12 to 13 µm long. Macroconidia germinated with a long germ tube before branching. These characteristics distinguished this fungus from M. laxa, a closely related species that is slow growing with lobed colony margins, produces few microconidia, and germ tubes that branch close to the conidium (1). The complete ITS sequence for DAOM 231119 was a 100% match to other sequences deposited for M. fructicola (Z73777, AF010500, and U21815). On the basis of comparisons of available data, ITS sequences for M. fructicola (three complete ITS, seven partial ITS) and M. laxa (8 complete ITS, 10 partial ITS) differed consistently at four nucleotide positions. The fungus identified as M. fructicola was tested for pathogenicity on mature surface-sterilized 'Pinot noir' and 'Riesling' grapes. Under humid conditions, buff-colored sporodochia bearing conidia developed over the surface of the infected berries. This indicates that M. fructicola can cause decay of wine grapes and could be confused with bunch rot caused by Botrytis cinerea. Previously, M. fructicola was reported on grapes in Oklahoma, but likely these grapes were not Vitis vinifera (2). To our knowledge, this is the first report of brown rot caused by M. fructicola on wine grapes in North America. References: (1) L. R. Batra. World Species of Monilinia (Fungi): Their Ecology, Biosystematics and Control. Mycologia Memoir No. 16. Gerbrüder Borntraeger, Berlin/Stuttgart, 1991. (2) D. A. Preston. Host Index of Oklahoma Plant Diseases, Tech. Bull. No. 21. Oklahoma Agricultural and Mechanical College, Agricultural Experiment Station, Stillwater, 1945.

Human eIF5A2 on chromosome 3q25-q27 is a phylogenetically conserved vertebrate variant of eukaryotic translation initiation factor 5A with tissue-specific expression.

Eukaryotic translation initiation factor 5A (eIF5A) is an essential protein tightly linked to cellular polyamine homeostasis. It receives the unique spermidine-derived posttranslational modification hypusine that is necessary for eIF5A's biochemical activity and cellular proliferation. The eIF5A protein stimulates ribosomal peptidyl-transferase and may be involved in nucleocytoplasmic mRNA transport. Little is known about the molecular genetics of eIF5A. Here we report on the sequence and molecular characterization of human EIF5A2, a novel phylogenetically conserved gene for eIF5A. EIF5A2 stretches over 17 kb and consists of five exons and four introns. It is localized at 3q25-q27, often noted for chromosomal instability in cancers. EIF5A2 is highly expressed in testis and colorectal adenocarcinoma and at moderate levels in the brain, in contrast to the ubiquitously expressed EIF5A1 gene. Two EIF5A2 mRNAs share a 129-nt 5' UTR and a coding sequence for the 153-amino-acid eIF5AII protein, but possess two alternative 3' UTRs of 46 and 890 nt that arise through differential polyadenylation. The protein is 84% identical and 94% similar to eIF5AI. Both EIF5A genes are conserved in vertebrates. Our findings lend further support for a specialized gene expression program of polyamine metabolic proteins and regulators that function to maintain polyamine homeostasis at elevated levels during spermatogenesis.

Evaluating the risk from depleted uranium after the Boeing 747-258F crash in Amsterdam, 1992.

On 4 October 1992, a large cargo plane crashed into an apartment building in the Bijlmermeer quarter of Amsterdam. In the years following the accident, an increasing number of people started reporting health complaints, which they attributed to exposure to dangerous substances after the crash. Since the aircraft had been carrying depleted uranium as counterbalance weights and about 150 kg uranium had been found missing after clearance of the crash site, exposure to uranium oxide particles was pointed out as the possible cause of their health complaints. Six years after the accident, a risk analysis was therefore carried out to investigate whether the health complaints could be attributed to exposure to uranium oxide set free during the accident. The scientific challenge was to come up with reliable results, knowing that - considering the late date - virtually no data were available to validate any calculated result. The source term of uranium was estimated using both generic and specific data. Various dispersion models were applied in combination with the local setting and the meteorological conditions at the time of the accident to estimate the exposure of bystanders during the fire caused by the crash. Emphasis was given to analysing the input parameters, inter-comparing the various models and comparing model results with the scarce information available. Uranium oxide formed in the fire has a low solubility, making the chemical toxicity to humans less important than the radiotoxicity. Best-estimate results indicated that bystanders may have been exposed to a radiation dose of less than 1 microSv, whereas a worst-case approach indicated an upper limit of less than 1 mSv. This value is considerably less than the radiation dose for which acute effects are to be expected. It is therefore considered to be improbable that the missing uranium had indeed led to the health complaints reported.

Effect of propentofylline (HWA 285) on focal ischemia in rats: effect of treatment and posttreatment duration on infarct size.

UNLABELLED: BACKGROUND AND PUROSEe: in this study we tested the potentially neuroprotective properties of propentofylline in a model of focal ischemia with long-term, repeated treatment. METHODS: 37 male Wistar rats (280-300 g) underwent permanent occlusion of the middle cerebral artery (MCA). Infusion was started 30 min after occlusion of the MCA over a period of 2 h with a dosage of 0.01 mg/kg body weight. Immediately after the termination of infusion repetitive intraperitoneal injections were started. Animals were assigned to four groups: continuous treatment for a period of 12 h with 24-h survival (group A, n=9) or 48-h survival (group B, n=10), continuous treatment for a period of 48 h with 48-h survival (group C, n=9) and placebo (group D, n=9). Infarct size was calculated from brain slices stained with 2,3,5-triphenyltetrazolium chloride. RESULTS: the infarct size was significantly reduced in group C (treatment for 48 h) (163.9+/-30.5 mm(3)) compared to the placebo group (297.4+/-17. 7 mm(3)). No effect on infarct size was observed in group A (196. 8+/-37.3 mm(3)) and group B (239.6+/-42.9 mm(3)) compared to placebo. CONCLUSION: continuous i.p. injections of propentofylline over a period of 48 h significantly reduces infarct size in an animal model of focal cerebral ischemia. With shorter periods of continuous administration of the drug and delayed postmortem analysis, reductions in the infarct size did not reach a level of significance. These data show the importance of continuous long-term administration after ischemic stroke in clinical trials to achieve the beneficial effects of neuroprotection by propentofylline.

[Late myelopathy after chemonucleolysis. Case report and review of the literature]. / Spätmyelopathie nach Chemonukleolyse. Fallbericht und Ubersicht über die Literatur.

Chemonucleolysis is a debated therapeutic method for herniated lumbar disc. We report a patient who suffered a sequence of characteristic sequels cumulating in late-onset myelopathy with persistent spastic paraplegia, sensory loss below T8 and bladder incontinence. Complications of chemonucleolysis are less frequent as compared to herniated disc surgery, but may cause severe impediment. Serious complications are anaphylactic shock, intracranial or spinal hemorrhage and transverse myelitis. This has to be taken into account for indication and patient information.

Two years of clinical experience with Procera titanium crowns.

PURPOSE: The purpose of the study were to evaluate the clinical performance of the Procera porcelain-fused-to-titanium crown system in general practice during a 2-year period, and to evaluate the performance of a new low-fusing porcelain as a veneering material on titanium. MATERIALS AND METHODS: A number of consecutive complete-coverage crowns (40) in 25 patients (14 women and 11 men) with a mean age of 53.1 years (range 35 to 79 years) were made according to the Procera (Nobel Biocare) technique in a 3-month period. The titanium copings were fabricated from solid rods of pure titanium using spark erosion and copy-milling technique, whereafter they were veneered with a new type of low-fusing ceramic material. The crowns were evaluated using the CDA criteria at baseline and after 2 years. RESULTS: The general failure rate was low and was restricted to one carious lesion, one porcelain fracture, and one loss of a crown resulting from failure of retention of a post and core. The most frequent single reason that the "excellent" color level was not recorded was a "too-high value." A slightly dull or granular porcelain surface was observed both at baseline and after 2 years. Overall, the responses of the patients were positive. CONCLUSION: Within the limitations of the study it can be concluded that porcelain-veneered Procera titanium crowns can be used as an alternative to other porcelain-fused-to-metal systems. However, conclusions should be made with caution from the results of this study because of the limited number of patients and crowns and the short observation period.