Identification of root canal microbiota profiles of periapical tissue diseases using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry.

OBJECTIVES: The purpose of this study was to identify microorganisms isolated from various periapical tissue diseases using Matrix-Assisted Laser Desorption Ionization Time-of-Flight Mass Spectrometry (MALDI-TOF-MS) and classify them via an unsupervised machine learning approach. METHODS: A total of 150 patients with various apical conditions and teeth in need of endodontic retreatment were divided into five groups, including Retreatment, Acute Apical Abscess, Chronic Apical Abscess, Acute Apical Periodontitis, and Chronic Apical Periodontitis. Samples were collected from root canals using paper points after agitating with a #10 K file then microorganisms were identified using MALDI-TOF-MS. Data were analyzed using a hierarchical clustering method. Quadruple clusters and dendrograms were formed according to similarities and dissimilarities. RESULTS: A total of 80 species were identified representative of six different phyla. The most similar microorganism species identified were: ''Enterococcus faecalis'' between 21 and 23-year-old female cases in Retreatment group; ''Lactobacillus rhamnosus'' between 20 and 18-year-old male cases in Symptomatic Apical Abscess cases; ''Lactobacillus paracasei'' between 26 and 40-year-old male cases in Asymptomatic Apical Abscess cases; ''Enterococcus faecalis'' between 48 and 50-year-old female cases in Symptomatic Apical Periodontitis cases; ''Lactobacillus rhamnosus'' between 48 and 60-year-old male cases in Asymptomatic Apical Periodontitis cases. CONCLUSIONS: MALDI-TOF MS can be considered a fast and high-throughput screening technique for microbial species identification in endodontics. Thus, it will provide valuable data for future research designs regarding periapical tissue diseases. As the MALDI-TOF MS database expands and comprehensive data becomes available, the relationship between microbial profiles and disease progression will become increasingly apparent.

Contemporary research trends on nanoparticles in endodontics: a bibliometric and scientometric analysis of the top 100 most-cited articles.

Objectives: Advancements in nanotechnology have led to the widespread usage of nanoparticles in the endodontic field. This bibliometric study aimed to determine and analyze the top 100 most-cited articles about nanoparticles in endodontics from 2000 to 2022. Materials and Methods: A detailed electronic search was conducted on the "Clarivate Analytics Web of Science, All Databases" to receive the most-cited articles related to the topic. Articles were ranked in descending order based on their citation counts, and the first 100 were selected for bibliometric analysis. Parameters such as citation density, publication year, journal, country, institution, author, study design, study field, evidence level, and keywords were analyzed. Results: The top 100 most-cited articles received 4,698 citations (16-271) with 970.21 (1.91-181) citation density in total. Among decades, citations were significantly higher in 2011-2022 (p < 0.001). Journal of Endodontics had the largest number of publications. Canada and the University of Toronto made the highest contribution as country and institution, respectively. Anil Kishen was the 1 who participated in the largest number of articles. The majority of the articles were designed in vitro. The main study field was "antibacterial effect." Among keywords, "nanoparticles" followed by "Enterococcus faecalis" were used more frequently. Conclusions: Developments in nanotechnology had an impact on the increasing number of studies in recent years. This bibliometric study provides a comprehensive view of nanoparticle advances and trends using citation analysis.

A G protein-coupled, IP3/protein kinase C pathway controlling the synthesis of phosphaturic hormone FGF23.

Dysregulated actions of bone-derived phosphaturic hormone fibroblast growth factor 23 (FGF23) result in several inherited diseases, such as X-linked hypophosphatemia (XLH), and contribute substantially to the mortality in kidney failure. Mechanisms governing FGF23 production are poorly defined. We herein found that ablation of the Gq/11α-like, extralarge Gα subunit (XLαs), a product of GNAS, exhibits FGF23 deficiency and hyperphosphatemia in early postnatal mice (XLKO). FGF23 elevation in response to parathyroid hormone, a stimulator of FGF23 production via cAMP, was intact in XLKO mice, while skeletal levels of protein kinase C isoforms α and δ (PKCα and PKCδ) were diminished. XLαs ablation in osteocyte-like Ocy454 cells suppressed the levels of FGF23 mRNA, inositol 1,4,5-trisphosphate (IP3), and PKCα/PKCδ proteins. PKC activation in vivo via injecting phorbol myristate acetate (PMA) or by constitutively active Gqα-Q209L in osteocytes and osteoblasts promoted FGF23 production. Molecular studies showed that the PKC activation-induced FGF23 elevation was dependent on MAPK signaling. The baseline PKC activity was elevated in bones of Hyp mice, a model of XLH. XLαs ablation significantly, but modestly, reduced serum FGF23 and elevated serum phosphate in Hyp mice. These findings reveal a potentially hitherto-unknown mechanism of FGF23 synthesis involving a G protein-coupled IP3/PKC pathway, which may be targeted to fine-tune FGF23 levels.

Comparison of Four Electronic Apex Locators in Detecting Working Length: An Ex Vivo Study.

AIM: To compare the accuracy of four different electronic apex locators (EALs) in detecting a position 0.5 mm short of the major foramen. MATERIALS AND METHODS: The actual working length of thirty-five extracted human teeth was determined visually as 0.5 mm short of the apical foramen. After actual working length measurements, electronic working length was measured with four different EALs (Apex Pointer+, Raypex 5, Apex ID, and Raypex 6). Measurements were repeated three times by different operators. The data were analyzed using the intraclass correlation coefficient (ICC), the repeated measure analysis of variance (rANOVA) and Bonferroni post hoc tests. The significance level was set at p < 0.05. RESULTS: The mean differences between electronic and actual working length were-0.305 mm, 0.098 mm, 0.037, and 0.144 mm for the Apex Pointer+, the Raypex 5, the Apex ID, and the Raypex 6, respectively. Multiple paired comparisons (Bonferroni test) also showed the Apex Pointer+ is significantly different from the Raypex 5, Apex ID and Raypex 6 (p = 0.000, p = 0.001, and p = 0.001 respectively). CONCLUSION: All EALs showed an acceptable determination of the working length between the ranges of ± 0.5mm except for the Apex Pointer+ device, which had the lowest accuracy. Further studies may be beneficial especially to better evaluate the accuracy of the Apex Pointer+. CLINICAL SIGNIFICANCE: This article shows that Apex ID, which has only recently been introduced into the market, showed an acceptable determination of the working length. Its accuracy was similar to that of Raypex 5 and 6.

Ablation of the Stimulatory G Protein α-Subunit in Renal Proximal Tubules Leads to Parathyroid Hormone-Resistance With Increased Renal Cyp24a1 mRNA Abundance and Reduced Serum 1,25-Dihydroxyvitamin D.

PTH regulates serum calcium, phosphate, and 1,25-dihydroxyvitamin D (1,25(OH)2D) levels by acting on bone and kidney. In renal proximal tubules (PTs), PTH inhibits reabsorption of phosphate and stimulates the synthesis of 1,25(OH)2D. The PTH receptor couples to multiple G proteins. We here ablated the α-subunit of the stimulatory G protein (Gsα) in mouse PTs by using Cre recombinase driven by the promoter of type-2 sodium-glucose cotransporter (Gsα(Sglt2KO) mice). Gsα(Sglt2KO) mice were normophosphatemic but displayed, relative to controls, hypocalcemia (1.19 ±0.01 vs 1.23 ±0.01 mmol/L; P < .05), reduced serum 1,25(OH)2D (59.3 ±7.0 vs 102.5 ±12.2 pmol/L; P < .05), and elevated serum PTH (834 ±133 vs 438 ±59 pg/mL; P < .05). PTH-induced elevation in urinary cAMP excretion was blunted in Gsα(Sglt2KO) mice (2- vs 4-fold over baseline in controls; P < .05). Relative to baseline in controls, PTH-induced reduction in serum phosphate tended to be blunted in Gsα(Sglt2KO) mice (-0.39 ±0.33 vs -1.34 ±0.36 mg/dL; P = .07). Gsα(Sglt2KO) mice showed elevated renal vitamin D 24-hydroxylase and bone fibroblast growth factor-23 (FGF23) mRNA abundance (∼3.4- and ∼11-fold over controls, respectively; P < .05) and tended to have elevated serum FGF23 (829 ±76 vs 632 ±60 pg/mL in controls; P = .07). Heterozygous mice having constitutive ablation of the maternal Gsα allele (E1(m-/+)) (model of pseudohypoparathyroidism type-Ia), in which Gsα levels in PT are reduced, also exhibited elevated serum FGF23 (474 ±20 vs 374 ±27 pg/mL in controls; P < .05). Our findings indicate that Gsα is required in PTs for suppressing renal vitamin D 24-hydroxylase mRNA levels and for maintaining normal serum 1,25(OH)2D.

Postnatal establishment of allelic Gαs silencing as a plausible explanation for delayed onset of parathyroid hormone resistance owing to heterozygous Gαs disruption.

Pseudohypoparathyroidism type-Ia (PHP-Ia), characterized by renal proximal tubular resistance to parathyroid hormone (PTH), results from maternal mutations of GNAS that lead to loss of α-subunit of the stimulatory G protein (Gαs) activity. Gαs expression is paternally silenced in the renal proximal tubule, and this genomic event is critical for the development of PTH resistance, as patients display impaired hormone action only if the mutation is inherited maternally. The primary clinical finding of PHP-Ia is hypocalcemia, which can lead to various neuromuscular defects including seizures. PHP-Ia patients frequently do not present with hypocalcemia until after infancy, but it has remained uncertain whether PTH resistance occurs in a delayed fashion. Analyzing reported cases of PHP-Ia with documented GNAS mutations and mice heterozygous for disruption of Gnas, we herein determined that the manifestation of PTH resistance caused by the maternal loss of Gαs, ie, hypocalcemia and elevated serum PTH, occurs after early postnatal life. To investigate whether this delay could reflect gradual development of paternal Gαs silencing, we then analyzed renal proximal tubules isolated by laser capture microdissection from mice with either maternal or paternal disruption of Gnas. Our results revealed that, whereas expression of Gαs mRNA in this tissue is predominantly from the maternal Gnas allele at weaning (3 weeks postnatal) and in adulthood, the contributions of the maternal and paternal Gnas alleles to Gαs mRNA expression are equal at postnatal day 3. In contrast, we found that paternal Gαs expression is already markedly repressed in brown adipose tissue at birth. Thus, the mechanisms silencing the paternal Gαs allele in renal proximal tubules are not operational during early postnatal development, and this finding correlates well with the latency of PTH resistance in patients with PHP-Ia.

The evaluation of two rotary instrumentation techniques under "operator-related variables" standardized conditions.

OBJECTIVE: We aimed here to evaluate the enlargement characteristics of two nickel-titanium rotary instrument systems that use two different preparation techniques, in simulated root canals under "operator-related variables" standardized conditions. MATERIALS AND METHODS: A total of 40 simulated canals in resin blocks were divided into two groups as the Mtwo group for use with a single-length technique and the ProTaper instrument system group for use with crown down technique. To standardize the operator-related variables, all preparations were carried out with a computer controlled device that was developed and used previously as described in a published study. The pre- and post-operative digital images of the canals were superimposed and measured at 11 levels from the apical tip. The distances between the pre- and post-operative outer lines of the root canals were measured at both the inner and outer sides of the curve. The amount of removed material and the symmetry of the preparations were evaluated. Statistical analyses were performed with Mann-Whitney test. RESULTS: Significantly more material was removed by Mtwo at 9 levels and by ProTaper at 2 levels (P < 0.05) while no significant difference was determined at 11 levels. The preparations made with ProTaper were more symmetrical at 4 levels while no significant difference was determined at 7 levels. CONCLUSION: Mtwo removed significantly more material than ProTaper at different levels of the curved root canals. Mtwo and ProTaper made symmetrical preparations and maintained the original shape of the root canal at different levels under controlled operator-related variables.

Solubility and apical sealing characteristics of a new calcium silicate-based root canal sealer in comparison to calcium hydroxide-, methacrylate resin- and epoxy resin-based sealers.

AIM: To assess and compare the water sorption, solubility and apical sealing ability of iRoot SP and three other widely used root canal sealers. MATERIALS AND METHODS: Solubility was assessed by immersing standardized samples of calcium silicate- (iRoot SP), calcium hydroxide- (Sealapex), methacrylate resin- (EndoREZ) and epoxy resin- (AH Plus) based sealers in distilled water and measuring weight gain and weight loss at 6 h, 24 h and daily for 14 days. Roots of extracted mandibular premolars (n = 80) were prepared with 0.04-taper nickel-titanium rotary files to a final size 40. Roots were then randomly divided into four experimental groups (n = 18) and two control groups (n = 4), root canal sealers were applied and apical leakage was assessed using the fluid filtration method. Data was analyzed using Kruskal Wallis analysis of variance and Mann-Whitney U-tests, with the level of significance set at p ≤ 0.05. RESULTS: EndoREZ exhibited the highest water sorption, followed by iRoot SP, Sealapex and AH Plus. Sealapex exhibited significantly higher solubility than the other sealers, whereas no significant differences in solubility levels were observed between the other three sealers tested. AH Plus exhibited significantly lower microleakage than Sealapex and EndoREZ, whereas no difference in microleakage was found between AH Plus and iRoot SP. CONCLUSIONS: In view of the study findings, all tested sealers except Sealapex met the ANSI/ADA's requirements for solubility and no difference was found between AH Plus and iRoot SP in terms of apical sealing ability.

A case of a benign cementoblastoma treated by enucleation and apicoectomy.

Cementoblastoma is a rare, benign, odontogenic neoplasm of ectomesenchymal origin, representing less than 6% of all odontogenic tumors. Despite its well-known typical features, there are still controversies regarding the management of the condition. This article presents the case of a benign cementoblastoma in a 17-year-old girl. The lesion was typical and associated with the mandibular right first molar. Endodontic treatment of the involved tooth, enucleation of the cementoblastoma, and apicoectomy of the affected roots were performed. Removal of the tumor while preserving the associated tooth resulted in normal osseous healing and no evidence of recurrence after one year. Based on the findings of the current report, it can be suggested that, in properly selected cases, it is possible to remove cementoblastomas that affect molars without extracting the involved teeth.

Comparison of cyclic fatigue resistance of three different rotary nickel-titanium instruments designed for retreatment.

INTRODUCTION: A number of rotary nickel-titanium (NiTi) systems have been developed to provide better, faster, and easier cleaning and shaping of the root canal system, and recently, rotary NiTi systems designed for root canal retreatment have been introduced. Because the main problem with the rotary NiTi files is fracture, the aim of this study was to compare the cyclic fatigue resistance of 3 different rotary NiTi systems designed for root canal retreatment. METHODS: Total of 60 instruments of 3 different rotary NiTi systems designed for root canal retreatment were used in this study. Twenty R-Endo R3, 20 ProTaper D3, and 20 Mtwo R (Retreatment) 25.05 instruments were tested. Cyclic fatigue testing of instruments was performed by using a device that allowed the instruments to rotate freely inside an artificial canal. Each instrument was rotated until fracture occurred, and the number of cycles to fracture for each instrument was calculated. Representative samples were also evaluated under a scanning electron microscope to confirm the fracture was flexural. Data were analyzed by using 1-way analysis of variance test. RESULTS: R-Endo R3 instruments showed better cyclic fatigue resistance than ProTaper D3 and Mtwo R 25.05 instruments, and the difference was statistically significant (P < .05). There was no significant difference between ProTaper D3 and Mtwo R 25.05 groups. CONCLUSIONS: The R-Endo R3 instruments were more resistant to fatigue failure than ProTaper D3 and Mtwo R 25.05.

Cyclic fatigue resistance of new and used Mtwo rotary nickel-titanium instruments in two different radii of curvature.

The fracture of nickel-titanium rotary instruments is a serious problem that can jeopardise the outcome of root canal treatment. The aim of this study was to compare the cyclic fatigue resistance of new and used Mtwo rotary nickel-titanium instruments in artificial canals with two different radii of curvature. Four hundred and twenty Mtwo instruments were used in the present study. Thirty new and 30 used instruments of each size were tested for cyclic fatigue in artificial canals with radii of curvature of 5 or 10 mm. Each instrument was rotated until fracture occurred and the number of cycles to fracture was calculated by multiplying time to fracture and rotational speed. Data were analysed using a t-test. According to the results of the study, radius of curvature was an important factor on cyclic fatigue resistance and clinical use significantly reduced the instrument lifespan.

Functional characterization of GNAS mutations found in patients with pseudohypoparathyroidism type Ic defines a new subgroup of pseudohypoparathyroidism affecting selectively Gsα-receptor interaction.

Pseudohypoparathyroidism type Ia (PHPIa) is caused by GNAS mutations leading to deficiency of the α-subunit of stimulatory G proteins (Gsα) that mediate signal transduction of G protein-coupled receptors via cAMP. PHP type Ic (PHPIc) and PHPIa share clinical features of Albright hereditary osteodystrophy (AHO); however, in vitro activity of solubilized Gsα protein is normal in PHPIc but reduced in PHPIa. We screened 32 patients classified as PHPIc for GNAS mutations and identified three mutations (p.E392K, p.E392X, p.L388R) in four unrelated families. These and one novel mutation associated with PHPIa (p.L388P) were introduced into a pcDNA3.1(-) expression vector encoding Gsα wild-type and expressed in a Gsα-null cell line (Gnas(E2-/E2-) ). To investigate receptor-mediated cAMP accumulation, we stimulated the endogenous expressed ß(2) -adrenergic receptor, or the coexpressed PTH or TSH receptors, and measured the synthesized cAMP by RIA. The results were compared to receptor-independent cholera toxin-induced cAMP accumulation. Each of the mutants associated with PHPIc significantly reduced or completely disrupted receptor-mediated activation, but displayed normal receptor-independent activation. In contrast, PHPIa associated p.L388P disrupted both receptor-mediated activation and receptor-independent activation. We present a new subgroup of PHP that is caused by Gsα deficiency and selectively affects receptor coupling functions of Gsα.

Transgenic overexpression of the extra-large Gsα variant XLαs enhances Gsα-mediated responses in the mouse renal proximal tubule in vivo.

XLαs, a variant of the stimulatory G protein α-subunit (Gsα), can mediate receptor-activated cAMP generation and, thus, mimic the actions of Gsα in transfected cells. However, it remains unknown whether XLαs can act in a similar manner in vivo. We have now generated mice with ectopic transgenic expression of rat XLαs in the renal proximal tubule (rptXLαs mice), where Gsα mediates most actions of PTH. Western blots and quantitative RT-PCR showed that, while Gsα and type-1 PTH receptor levels were unaltered, protein kinase A activity and 25-hydroxyvitamin D 1-α-hydroxylase (Cyp27b1) mRNA levels were significantly higher in renal proximal tubules of rptXLαs mice than wild-type littermates. Immunohistochemical analysis of kidney sections showed that the sodium-phosphate cotransporter type 2a was modestly reduced in brush border membranes of male rptXLαs mice compared to gender-matched controls. Serum calcium, phosphorus, and 1,25 dihydroxyvitamin D were within the normal range, but serum PTH was ∼30% lower in rptXLαs mice than in controls (152 ± 16 vs. 222 ± 41 pg/ml; P < 0.05). After crossing the rptXLαs mice to mice with ablation of maternal Gnas exon 1 (E1(m-/+)), male offspring carrying both the XLαs transgene and maternal Gnas exon 1 ablation (rptXLαs/E1(m-/+)) were significantly less hypocalcemic than gender-matched E1(m-/+) littermates. Both E1(m-/+) and rptXLαs/E1(m-/+) offspring had higher serum PTH than wild-type littermates, but the degree of secondary hyperparathyroidism tended to be lower in rptXLαs/E1(m-/+) mice. Hence, transgenic XLαs expression in the proximal tubule enhanced Gsα-mediated responses, indicating that XLαs can mimic Gsα in vivo.

Potent constitutive cyclic AMP-generating activity of XLαs implicates this imprinted GNAS product in the pathogenesis of McCune-Albright syndrome and fibrous dysplasia of bone.

Patients with McCune-Albright syndrome (MAS), characterized primarily by hyperpigmented skin lesions, precocious puberty, and fibrous dyslasia of bone, carry postzygotic heterozygous mutations of GNAS causing constitutive cAMP signaling. GNAS encodes the α-subunit of the stimulatory G protein (Gsα), as well as a large variant (XLαs) derived from the paternal allele. The mutations causing MAS affect both GNAS products, but whether XLαs, like Gsα, can be involved in the pathogenesis remains unknown. Here, we investigated biopsy samples from four previously reported and eight new patients with MAS. Activating mutations of GNAS (Arg201 with respect to the amino acid sequence of Gsα) were present in all the previously reported and five of the new cases. The mutation was detected within the paternally expressed XLαs transcript in five and the maternally expressed NESP55 transcript in four cases. Tissues carrying paternal mutations appeared to have higher XLαs mRNA levels than maternal mutations. The human XLαs mutant analogous to Gsα-R201H (XLαs-R543H) showed markedly higher basal cAMP accumulation than wild-type XLαs in transfected cells. Wild-type XLαs demonstrated higher basal and isoproterenol-induced cAMP signaling than Gsα and co-purified with Gß1γ2 in transduced cells. XLαs mRNA was measurable in mouse calvarial cells, with its level being significantly higher in undifferentiated cells than those expressing preosteoblastic markers osterix and alkaline phosphatase. XLαs mRNA was also expressed in murine bone marrow stromal cells and preosteoblastic MC3T3-E1 cells. Our findings are consistent with the possibility that constitutive XLαs activity adds to the molecular pathogenesis of MAS and fibrous dysplasia of bone.

Dislocation resistance of iRoot SP, a calcium silicate-based sealer, from radicular dentine.

INTRODUCTION: The aim of this study was to evaluate the push-out bond strength of iRoot SP (Innovative Bioceramix, Vancouver, Canada) and compare it with that of other widely used root canal sealers. METHODS: Sixty extracted human maxillary canines were sectioned transversally below the cement-enamel junction to obtain 120 4-mm-thick dentin disks that were randomly divided into four groups (n = 30) for treatment with one of four different root-canal sealers (iRoot SP, AH Plus [Dentsply DeTrey GmbH, Konstanz, Germany], Sealapex [SybronEndo Corporation, Orange, CA], EndoREZ [Ultradent Inc, South Jordan, UT]). Standardized cavities were prepared to simulate root canals, cavities were filled with sealer material, and push-out bond-strength testing was performed using a universal testing machine. Failure modes were assessed quantitatively under a stereomicroscope and morphologically under a scanning electron microscope. Statistical analysis was performed using one-way analysis of variance and the post hoc Tukey test, with the significance level set at 0.05. RESULTS: Bond strengths of iRoot SP and AH Plus were significantly higher than those of Sealapex and EndoREZ. There was no significant difference between the bond strength of iRoot SP and AH Plus (p = 0.274). CONCLUSIONS: iRoot SP and AH Plus performed similarly and better than EndoREZ and Sealapex in terms of bond strength.

Is endodontic treatment necessary during coronectomy procedure?

PURPOSE: Close proximity of the inferior alveolar nerve (IAN) to the third molar roots can result in nerve injuries during extraction of third molars. Consequently, it is necessary to determine the relationship of the nerve and roots to avoid damage to the IAN. Computed tomography scans are widely used to determine the correct relationship between the IAN and lower third molars. PATIENTS AND METHODS: The study consisted of 10 patients with 16 lower third molars in close relationship with the IAN who were divided into a study group and a control group. The patients in the study group were treated via coronectomies performed with endodontic treatments. The patients in the control group underwent coronectomies without endodontic treatment. The patients were followed up for at least 1 year. RESULTS: We had to extract 7 of the roots because of the infection in 8 patients belonging to the study group, which were treated endodontically. Moreover, there were 3 cases of IAN damage because of the extraction in the study group. However, in the control group, no infection was determined and IAN damage was absent. CONCLUSION: Coronectomy appears to be a reliable technique to protect the IAN from damage. This procedure has a low incidence of complications. Endodontic treatment does not affect the success of this method according to our results.

Comparison of cyclic fatigue resistance of used and new RaCe instruments.

OBJECTIVE: The aim of this study was to evaluate cyclic fatigue resistance of used and new RaCe rotary nickel-titanium instruments. STUDY DESIGN: Forty RaCe (FKG Dentaire, La-Chaux-de-Fonds, Switzerland) instruments of sizes 30/.06, 30/.02, 25/.04, and 25/.02 were selected and divided into 2 groups: group A = 20 new instruments; group B = 20 used instruments. In group B, each instrument was used for shaping 5 curved molar root canals. Cyclic fatigue testing of new and used instruments was performed using a device that allowed the instruments to rotate freely inside a stainless steel artificial canal. Data were analyzed using t test to determine any statistical significance between used and new instruments of the same file size. RESULTS: For all sizes, new instruments were more resistant to failure then the used ones (P < .05). A reduction in lifespan was observed ranging from 18% (30/.06) to 51% (25/.02). CONCLUSION: The results showed that clinical use significantly reduced cyclic fatigue resistance of RaCe rotary instruments compared with new ones.

Identification of a novel dentin matrix protein-1 (DMP-1) mutation and dental anomalies in a kindred with autosomal recessive hypophosphatemia.

An autosomal recessive form of hypophosphatemia (ARHP) was recently shown to be caused by homozygous mutations in DMP1, the gene encoding dentin matrix protein-1 (DMP-1), a non-collagenous bone matrix protein with an important role in the development and mineralization of bone and teeth. Here, we describe a previously not reported consanguineous ARHP kindred in which the three affected individuals carry a novel homozygous DMP-1 mutation. The index case presented at the age of 3 years with bowing of his legs and showed hypophosphatemia due to insufficient renal phosphate retention. Serum alkaline phosphatase activity was elevated, with initially normal PTH. FGF23 was inappropriately normal at an older age while being treated with oral phosphate and 1,25(OH)(2)D. Similar clinical and biochemical findings, except for elevated FGF23 levels, were present in his 16-month-old brother and his 12.5-year-old female cousin; the parents of the three affected children are first-degree cousins. Nucleotide sequence analysis was performed on PCR-amplified exons encoding DMP-1 and flanking intronic regions. A novel homozygous frame-shift mutation (c.485Tdel; p.Glu163ArgfsX53) in exon 6 resulting in a premature stop codon was identified in all effected individuals. The parents and available unaffected siblings were heterozygous for c.485Tdel. Tooth growth and shape were normal for the index case, his affected brother and cousin, but their permanent and deciduous teeth displayed enlarged pulp chambers. The identified genetic mutation underscores the importance of DMP-1 mutations in the pathogenesis of ARHP. Furthermore, DMP-1 mutations appear to contribute, through yet unknown mechanisms, to tooth development.

Extralarge XL(alpha)s (XXL(alpha)s), a variant of stimulatory G protein alpha-subunit (Gs(alpha)), is a distinct, membrane-anchored GNAS product that can mimic Gs(alpha).

GNAS gives rise to multiple imprinted gene products, including the alpha-subunit of the stimulatory G protein (Gs(alpha)) and its variant XL(alpha)s. Based on genomic sequence, the translation of XL(alpha)s begins from the middle of a long open reading frame, suggesting the existence of an N-terminally extended variant termed extralarge XLalphas (XXL(alpha)s). Although XXL(alpha), like Gs(alpha) and XL(alpha)s, would be affected by most disease-causing GNAS mutations, its authenticity and biological significance remained unknown. Here we identified a mouse cDNA clone that comprises the entire open reading frame encoding XXL(alpha)s. Whereas XXL(alpha)s mRNA was readily detected in mouse heart by RT-PCR, it appeared virtually absent in insulinoma-derived INS-1 cells. By Northern blots and RT-PCR, XXL(alpha)s mRNA was detected primarily in the mouse brain, cerebellum, and spleen. Immunohistochemistry using a specific anti-XXL(alpha)s antibody demonstrated XXL(alpha)s protein in multiple brain areas, including dorsal hippocampus and cortex. In transfected cells, full-length human XXL(alpha)s was localized to the plasma membrane and mediated isoproterenol- and cholera toxin-stimulated cAMP accumulation. XXL(alpha)s-R844H, which bears a mutation analogous to that in the constitutively active Gs(alpha) mutant Gs(alpha)-R201H (gsp oncogene), displayed elevated basal signaling. However, unlike Gs(alpha)-R201H, which mostly remains in the cytoplasm, both XXL(alpha)s-R844H and a constitutively active XL(alpha)s mutant localized to the plasma membrane. Hence, XXL(alpha)s is a distinct GNAS product and can mimic Gs(alpha), but the constitutively active XXL(alpha)s and Gs(alpha) mutants differ from each other regarding subcellular targeting. Our findings suggest that XXL(alpha)s deficiency or hyperactivity may contribute to the pathogenesis of diseases caused by GNAS mutations.

In vitro evaluation of matched-taper single-cone obturation with a fluid filtration method.

OBJECTIVE: Widespread use of rotary nickel-titanium systems has caused manufacturers to produce gutta-percha cones that match the taper of the canals prepared with these systems. Obturations with these cones may provide 3-dimensional obturation of the root canal in less time. The aim of this study was to use a fluid filtration method to compare the apical sealing ability of matched-taper single-cone, Thermafil and lateral condensation obturations. MATERIALS AND METHODS: Sixty-six extracted human lower premolar teeth were instrumented to size F3 with ProTaper instruments. The teeth were divided into 3 groups and obturated with matched-taper single-cone, Thermafil and lateral condensation techniques. AH Plus was used as a sealer. Six teeth were used as positive and negative controls. A fluid filtration system was used to evaluate the apical seal. Measurements were made at 2-minute intervals for 8 minutes and the average of these values was recorded. One-way analysis of variance test was used for statistical analysis. RESULTS: Mean leakage values for the lateral condensation, single-cone and Thermafil groups were 0.49 +/- 0.37 microL min(-1), 0.39 +/- 0.24 microL min(-1) and 0.44 +/- 0.25 microL min(-1), respectively. The difference between the groups was not statistically significant (p > 0.05). CONCLUSIONS: The apical sealing ability of matched-taper single-cone obturation was comparable with that of lateral condensation and Thermafil techniques.