Canal-Centering and Apical Transportation Ability of Similar Cross-Section NiTi Instruments Working with Different Kinematics-Micro-CT-based In Vitro Analysis.

Objective: Evaluating the canal-centering and apical transportation ability of endodontic file systems working with different kinematics but of similar cross section. Materials and Methods: Sixty human maxillary first molar mesiobuccal (MB) roots were assigned to three experimental groups based on instrumentation techniques: Reciproc Blue (RB), Mtwo (M2), and OneShape (OS). Pre- and post-instrumentation micro-computed tomographic analysis was performed. Centering ability and apical transportation were analyzed at 3 mm, 6 mm, and 9 mm short of the apex. Statistical analysis was conducted using Mann-Whitney U test and Kruskal-Wallis test. Results: OS showed better canal-centering ability than RB and M2 at 3 mm, 6 mm, and 9 mm. No significant difference among the tested groups was observed during the assessment of apical transportation (P < 0.05). Conclusion: The systems evaluated safely prepared curved MB canals with minimal canal transportation. OS showed superior canal-centering ability compared to the other two groups.

Effect of a novel quaternary ammonium silane based cavity cleanser FiteBac 2% K21 QAS in comparison with other cavity disinfectants on the bond strength of resin-modified glass ionomer cement.

Background: The application of cavity cleansers for cavity disinfection can be a crucial step in the longevity of restorations. The objective of the present study was to compare the effect of the application of a new quaternary ammonium silane (QAS)-based cavity cleanser (2% K21 QAS), with other commercially available cavity disinfectants on the bond strength of resin-modified glass ionomer cement (RMGIC). Materials and Methods: The buccal surfaces of 40 extracted premolars were trimmed to obtain a flat dentinal surface and were randomly divided into four experimental groups depending on the cavity cleansers used before restoration. Group 1: 2% chlorhexidine (CHX), Group 2: QAS (FiteBac 2% K21 QAS), Group 3: silver diamine fluoride-potassium iodide (Riva Star, SDF-KI), and Group 4: 3% hydrogen peroxide (H2O2). Then, a predetermined dimension of RMGIC restoration was bonded to the treated dentin surfaces. Following this, each sample was tested for shear bond strength (SBS) using a universal testing machine at a crosshead speed of 0.5 mm/min. Results: Among the experimental groups, SDF-KI has shown the highest mean SBS, followed by 2% K21 QAS, and 2% CHX, which have shown almost comparable results. The 3% H2O2 group has shown the lowest values. Conclusion: According to the results of the present study, 2% K21 QAS has the potential to be used as an effective cavity cleanser before the placement of RMGIC restorations. Since its application does not affect the bond strength of restoration, it can be successfully used as an alternative to CHX and SDF-KI.

Targeted Endodontic Microsurgery: A Guided Approach - A Report of Two Cases.

Targeted endodontic microsurgery combines a precisely designed three-dimensional (3D)-printed surgical guide in which the osteotomy site and angulation is defined preoperatively to avoid damaging anatomically important structures. The current endodontic microsurgical procedures have been progressing in pace with technological advances as a predictable alternative to nonsurgical treatment of persistent and recurrent apical periodontitis. The 3D-printed template has been used earlier in the guided endodontic procedure (access openings). The endodontic microsurgery utilizes the surgical microscope and microsurgical instruments which help in enhanced magnification, illumination, and visualization compared to conventional endodontic surgery. Cone beam computed tomography (CBCT) plays an important role in surgical endodontics as it helps in measuring the distance between the cortical plate and the apex, position of the roots within the bone, and the proximity of vital structures can be assessed. The true size, location, and extent of the periapical lesion can also be appreciated preoperatively. In the present cases, the guide allowed the clinicians to precisely reach the targeted tissues in a faster and more accurate manner with a more conservative and less traumatic treatment procedure. A 1-year CBCT follow-up of both cases showed complete 3D healing of the surgical site.

Transition of Methotrexate Polyglutamate Drug Monitoring Assay from Venipuncture to Capillary Blood-Based Collection Method in Rheumatic Diseases.

OBJECTIVE: Methotrexate (MTX) polyglutamate (MTXPG3) levels from isolated red blood cells (RBCs) collected by venipuncture have clinical utility in guiding MTX dosing for patients with rheumatoid arthritis (RA). Our objective was to transition this RBC-based therapeutic drug monitoring (TDM) assay to dried capillary blood collected by fingerstick. METHODS: Patients with RA treated with MTX were enrolled. Specimens were collected by fingerstick (volumetric absorptive microsampler) and venipuncture to measure MTXPG3 from dried capillary blood, total venous blood, and isolated RBCs. MTXPG3 levels from dried capillary blood were measured using LC-MS/MS, converted to RBC equivalent (nmol/L), and compared with those from isolated RBCs (reference method). Following transition to fingerstick collection, comparability in the distributions of dried capillary and venipuncture-based RBC MTXPG3 levels was assessed using the Kolmogorov-Smirnov (K-S) test. RESULTS: Intraday and interday precision ranged from 2.0% to 10.9% and 3.1% to 10.8%, respectively, at MTXPG3 concentrations ranging from 5 to 100 nmol/L. In 106 participants treated with MTX, MTXPG3 levels from total venous and dried capillary blood were comparable [slope = 0.97 (95% CI, 0.92-1.03); R 2 = 0.92]. Dried capillary blood MTXPG3 converted to RBC equivalent was similar to levels from isolated RBCs (30 ± 18 nmol/L vs 33 ± 19 nmol/L; n = 106). After implementation in the clinical laboratory, RBC equivalents MTXPG3 from the fingerstick method were similar to levels from venipuncture [39 ± 22 nmol/L (n = 825) vs 39 ± 24 nmol/L (n = 47935)] (K-S test P = 0.09). Underexposure to MTX (MTXPG3 ≤5 nmol/L RBCs) was detected in 7.0% and 8.5% patient specimens collected using the fingerstick and venipuncture methods, respectively. CONCLUSION: Capillary blood MTXPG3 levels can be used to guide MTX dosing in TDM practice.

A clinical correlation between stature and posterior tooth length.

INTRODUCTION: Exploration and determination of the relationship between stature and length of tooth is essential in Paleontology, Forensic Odontology and Endodontology. This study aimed to determine any association between stature and posterior tooth length in a group of patients who required root canal treatment. METHODS: Age, sex and standing height of adults were considered for posterior tooth length measurement. Molars and Premolars of apparently normal males (n=115 for molars, n= 75 for premolars) and females (n=124 for molars, n=80 for premolars), aged 20-50 years with intact cuspal morphology, which required RCT were selected for this study. Females and males were divided into 2 groups each based on their heights females > 155 cm and ≤ 155 cm, males > 165.10 and ≤ 165.10cm. The tooth length of permanent molars and premolars in both groups were measured using RVG and Electronic apex locator. Measurements obtained were compared separately for males and females using descriptive statistics and Pearson correlation coefficient. RESULTS: In females MB, ML, D roots of molar showed significant association (P=0.021), (P=0.027), (P=0.010) and roots of premolars showed significant association (P=0.002), (P=0.006) between both the groups respectively In males MB, ML, D roots of molar showed significant association (P=0.009), (P=0.004), (P=0.015) and roots of premolars showed significant association (P=0.006), (P=0.020) between both the groups respectively. CONCLUSION: The present clinical study reveals that there is a positive association between stature and posterior tooth length in both males and females.

Capillary blood collected on volumetric absorptive microsampling (VAMS) device for monitoring hydroxychloroquine in rheumatoid arthritis patients.

A novel technique for collection of capillary blood, termed volumetric absorptive microsampling (VAMS), has been recently cleared by the FDA for collection of human blood. VAMS absorbs a fixed volume of blood (10µl) and overcomes area bias and homogeneity issues associated with dried blood spot (DBS). This study is the application of VAMS for therapeutic drug monitoring (TDM) in human capillary blood. A liquid chromatography-tandem mass spectrometry (LC-MS/MS) workflow for analysis of VAMS sample was developed and validated. Concentration of hydroxychloroquine (HCQ) and its metabolites, desethylhydroxychloroquine (DHCQ), desethylchloroquine (DCQ), and bisdesethylchloroquine (BDCQ), in capillary blood on VAMS sampler were compared to those in venous blood in rheumatoid arthritis patients. Feasibility of capillary blood collected on both VAMS and DBS card were evaluated on patients. Stability of dried capillary blood on VAMS was also examined. Our results established that VAMS is a simple and accurate sampling technique that delivers the benefits of DBS sampling while overcoming the issues associated with hematocrit and homogeneity. It requires a small blood volume, simplifies sample logistics management, and may allow sample collection in the patient's home setting.

Resin bond strength to water versus ethanol-saturated human dentin pretreated with three different cross-linking agents.

CONTEXT: Resin-dentin bonds are unstable owing to hydrolytic and enzymatic degradation. Several approaches such as collagen cross-linking and ethanol-wet bonding (EWB) have been developed to overcome this problem. Collagen cross-linking improves the intrinsic properties of the collagen matrix. However, it leaves a water-rich collagen matrix with incomplete resin infiltration making it susceptible to fatigue degradation. Since EWB is expected to overcome the drawbacks of water-wet bonding (WWB), a combination of collagen cross-linking with EWB was tested. AIM: The aim of this study was to compare the effect of pretreatment with different cross-linking agents such as ultraviolet A (UVA)-activated 0.1% riboflavin, 1 M carbodiimide, and 6.5 wt% proanthocyanidin on the immediate and long-term bond strengths of an etch and rinse adhesive system to water- versus ethanol-saturated dentin within clinically relevant application time periods. SETTINGS AND DESIGN: Long-term in vitro study evaluating the microtensile bond strength of adhesive-dentin interface after different surface pretreatments. SUBJECTS AND METHODS: Eighty freshly extracted human molars were prepared to expose dentin, etched with 37% phosphoric acid for 15 s rinsed, and grouped randomly. They were blot-dried and pretreated with different cross-linkers: 0.1% riboflavin for 2 min followed by UVA activation for 2 min; 1 M carbodiimide for 2 min; 6.5 wt% proanthocyanidin for 2 min and rinsed. They were then bonded with Adper Single Bond Adhesive (3M ESPE), by either WWB or EWB, followed by resin composite build-ups (Filtek Z350, 3M ESPE). Bonded specimens in each group were then sectioned and divided into two halves. Microtensile bond strength was tested in one half after 24 h and the other after 6 months storage in artificial saliva. STATISTICAL ANALYSIS USED: Analysis was done using SPSS version 18 software (SPSS Inc., Chicago, IL, USA). Intergroup comparison of bond strength was done using ANOVA with post hoc Tukey's test, and intragroup comparison was done using paired t-test. RESULTS: The microtensile bond strength of cross-linked groups was higher compared to control group (P < 0.001). EWB showed much higher bond strength values on cross-linked dentin compared to noncross-linked dentin. UVA-activated riboflavin group exhibited highest bond strengths followed by carbodiimide and proanthocyanidin groups, respectively, on both water- as well as ethanol-saturated dentin. Even after 6 months storage, cross-linked groups showed significantly higher values compared to initial bond strength values of control group (P < 0.001). CONCLUSIONS: 0.1% riboflavin pretreatment of dentin followed by UVA activation for 2 min exhibited highest increase in bond strength values at 24 h and least reduction in bond strength values after 6 months storage compared to other groups. Biomodification of dentin using collagen cross-linking followed by EWB exhibited a synergistic effect in improving the resin-dentin bond durability.

Endodontic management of a mandibular first molar with six root canal systems.

Internal anatomy of pulp is complex. The first mandibular molars typically have two roots, one mesial with two root canals and another distal root, which contains one or two canals. A 20-year-old female patient reported with intermittent pain and incomplete root canal treatment in left lower back region since 1-week. Refined access cavity revealed initially two canals in mesial and two canals in the distal root. With operating microscope and cone beam computerized tomography, two additional canals (L-mesio-buccal and B-mesio-lingual) were identified in mesial root. One-year follow-up showed patient was asymptomatic and complete healing of periapical radiolucency.

Combining anti-ERBB3 antibodies specific for domain I and domain III enhances the anti-tumor activity over the individual monoclonal antibodies.

BACKGROUND: Inappropriate signaling through the epidermal growth factor receptor family (EGFR1/ERBB1, ERBB2/HER2, ERBB3/HER3, and ERBB4/HER4) of receptor tyrosine kinases leads to unregulated activation of multiple downstream signaling pathways that are linked to cancer formation and progression. In particular, ERBB3 plays a critical role in linking ERBB signaling to the phosphoinositide 3-kinase and Akt signaling pathway and increased levels of ERBB3-dependent signaling is also increasingly recognized as a mechanism for acquired resistance to ERBB-targeted therapies. METHODS: We had previously reported the isolation of a panel of anti-ERBB3 single-chain Fv antibodies through use of phage-display technology. In the current study scFv specific for domain I (F4) and domain III (A5) were converted into human IgG1 formats and analyzed for efficacy. RESULTS: Treatment of cells with an oligoclonal mixture of the A5/F4 IgGs appeared more effective at blocking both ligand-induced and ligand-independent signaling through ERBB3 than either single IgG alone. This correlated with improved ability to inhibit the cell growth both as a single agent and in combination with other ERBB-targeted therapies. Treatment of NCI-N87 tumor xenografts with the A5/F4 oligoclonal led to a statistically significant decrease in tumor growth rate that was further enhanced in combination with trastuzumab. CONCLUSION: These results suggest that an oligoclonal antibody mixture may be a more effective approach to downregulate ERBB3-dependent signaling.

Tailor-made endodontic obturator for the management of Blunderbuss canal.

The complex anatomy of the blunderbuss root canal often poses a major challenge to accomplish adequate obturation for a biological seal. Moreover, the roll-cone, Gutta-percha obturation technique, which is routinely practiced, also results in a mismatch and failure to configure to the canal volume in the absence of an apical barrier. Hence, an attempt has been made to tailor-make a heat polymerized polymethyl methacrylate resin as an endodontic obturator, to match the canal volume, which has been ascertained by Spiral computed tomography and mathematical integration. A one-year follow-up examination has revealed that the tooth is asymptomatic, with the repair of the lesion evident radiographically.

Evaluation of the anti-HER2 C6.5 diabody as a PET radiotracer to monitor HER2 status and predict response to trastuzumab treatment.

PURPOSE: The rapid tumor targeting and pharmacokinetic properties of engineered antibodies make them potentially suitable for use in imaging strategies to predict and monitor response to targeted therapies. This study aims to evaluate C6.5 diabody (C6.5 db), a noncovalent anti-HER2 single-chain Fv dimer, as a radiotracer for predicting response to HER2-targeted therapies such as trastuzumab. EXPERIMENTAL DESIGN: Immunodeficient mice bearing established HER2-positive tumor xenografts were injected with radioiodinated C6.5 db and imaged by PET/CT. Radiotracer biodistribution was quantified by biopsied tumor and normal tissues. Potential competition between trastuzumab and C6.5 db was examined in vitro by flow cytometry and coimmunoprecipitations. RESULTS: Biodistribution analysis of mice bearing xenografts with varying HER2 density revealed that the tumor uptake of (125)I-C6.5 db correlates with HER2 tumor density. In vitro competition experiments suggest that the C6.5 db targets an epitope on HER2 that is distinct from that bound by trastuzumab. Treatment of mice affected with SK-OV-3 tumor with trastuzumab for 3 days caused a 42% (P = 0.002) decrease in tumor uptake of (125)I-C6.5 db. This is consistent with a dramatic decrease in the tumor PET signal of (124)I-C6.5 db after trastuzumab treatment. Furthermore, mice affected with BT-474 tumor showed an approximately 60% decrease (P = 0.0026) in C6.5 db uptake after 6 days of trastuzumab treatment. Immunohistochemistry of excised xenograft sections and in vitro flow cytometry revealed that the decreased C6.5 db uptake on trastuzumab treatment is not associated with HER2 downregulation. CONCLUSIONS: These studies suggest that (124)I-C6.5 db-based imaging can be used to evaluate HER2 levels as a predictor of response to HER2-directed therapies.

Immuno-positron emission tomography in cancer models.

Positron emission tomography (PET) is playing an increasingly important role in the diagnosis, staging, and monitoring response to treatment in a variety of cancers. Recent efforts have focused on immuno-PET, which uses antibody-based radiotracers, to image tumors based on expression of tumor-associated antigens. It is postulated that the specificity afforded by antibody targeting should both improve tumor detection and provide phenotypic information related to primary and metastatic lesions that will guide therapy decisions. Advances in antibody-engineering are providing the tools to develop antibody-based molecules with pharmacokinetic properties optimized for use as immuno-PET radiotracers. Coupled with technical advances in the design of PET scanners, immuno-PET holds promise to improve diagnostic imaging and to guide the use of targeted therapies. An overview of the preclinical immuno-PET studies in cancer models is reviewed here.

In vivo role of a potassium channel-binding protein in regulating neuronal excitability and behavior.

Molecular details of ion channel interactions with modulatory subunits have been investigated widely in transfected cells, but the physiological roles of ion channel modulatory protein complexes in native neurons remain largely unexplored. The Drosophila large-conductance calcium-activated potassium channel (dSlo) binds to and is modulated by its binding partner Slob. We have constructed flies in which Slob expression is manipulated by P-element mutagenesis, or by transgenic expression of Slob protein or Slob-RNAi. In vivo recordings of both macroscopic and single dSlo channel currents in identified neurosecretory neurons in the pars intercerebralis (PI) region of the Drosophila brain reveal that whole-cell potassium current and properties of single dSlo channels are modulated by Slob expression level. Furthermore, Slob genotype influences action potential duration in vivo. This unprecedented combination of current-clamp, macroscopic-current, and single-channel recordings from neurons in brains of living flies defines a critical role for an ion channel modulatory protein complex in the control of neuronal excitability. We show further that Slob-null flies exhibit significantly longer lifespan than controls under conditions of complete food deprivation. Crosses with deficiency lines demonstrate that this enhanced resistance to starvation-induced death maps close to the slob locus. Together, these results indicate that Slob may serve a novel regulatory function in feeding behavior, possibly by influencing the excitability of the PI neurons.

Monomeric 14-3-3 protein is sufficient to modulate the activity of the Drosophila slowpoke calcium-dependent potassium channel.

Drosophila 14-3-3zeta (D14-3-3zeta) modulates the activity of the Slowpoke calcium-dependent potassium channel (dSlo) by interacting with the dSlo binding protein, Slob. We show here that D14-3-3zeta forms dimers in vitro. Site-directed mutations in its putative dimerization interface result in a dimerization-deficient form of D14-3-3zeta. Both the wild-type and dimerization-deficient forms of D14-3-3zeta bind to Slob with similar affinity and form complexes with dSlo. When dSlo and Slob are expressed in mammalian cells, the dSlo channel activity is similarly modulated by co-expression of either the wild-type or the dimerization-deficient form of D14-3-3zeta. In addition, dSlo is still modulated by wild-type D14-3-3zeta in the presence of a 14-3-3 mutant, which does not itself bind to Slob but forms heterodimers with the wild-type 14-3-3. These data, taken together, suggest that monomeric D14-3-3zeta is capable of modulating dSlo channel activity in this regulatory complex.