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AIM To determine whether irisin, a newly discovered myokine that links exercise-induced and metabolic homeostasis, is able to promote odontogenic differentiation and angiogenesis in human dental pulp cells (HDPCs). METHODOLOGY Cell viability in the presence of irisin was measured. Real-time PCR and Western blot analysis were performed to evaluate the expression levels of irisin, odontogenic and angiogenic markers. The involvement of mitogen-activated protein kinase (MAPK) and the protein kinase B (Akt) signalling pathway was evaluated by Western blot. To evaluate mineralization nodule formation, alkaline phosphatase (ALP) staining and alizarin red S staining were performed. Scratch wound assays were performed to evaluate the effects of irisin on cell migration. The data were analysed using one-way analysis of variance (anova) followed by Tukey post hoc test and Student's t-test. Statistical significance was considered at P

Objective: The aim of this study was to compare the characteristics of periodontal ligament stem cells derived from supernumerary teeth (sPDLSCs), cultured using a three-dimensional (3D) method and a conventional two-dimensional (2D) method. Methods: The morphology, viability, and osteogenic differentiation of the cells were analyzed. In addition, gene expression was analyzed by RNA sequencing, to characterize the functional differences. Results: The diameter of the 3D-cultured sPDLSCs decreased over time, but the spheroid shape was maintained for 7 days. The osteogenic differentiation was similar in the 2D and 3D. The gene expression related to the extracellular matrix (7.3%), angiogenesis (5.6%), cell proliferation (4.6%), inflammatory response (3.7%), and cell migration (3.5%) differed (p &lt, 0.05). Conclusions: Within the limitations of this study, sPDLSCs varied in formation and function, depending on the culture method. In future, it is necessary to study tissue engineering using the advantages of 3D culture and the fewer ethical problems of supernumerary teeth.

INTRODUCTION Leptin is secreted as a peptide hormone from adipose tissues. The aim of this study was to evaluate the effects of leptin on reparative dentin formation and angiogenesis in the pulp tissue of teeth in vivo. METHODS Twenty-four 7-week-old male rats were anesthetized. Cavities were prepared in maxillary first molars. Pulp cappings were performed with collagen scaffold (Col) with a phosphate-buffered saline (PBS) vehicle (Col + PBS), leptin 1 μmol/L with Col (L1 + Col), or leptin 10 μmol/L with Col (L10 + Col). For the negative control group (no pulp capping), pulp capping was not performed. All cavities were sealed with resin-modified glass ionomer followed by a micro-computed tomographic scan, histologic examination, and immunohistochemical analysis. RESULTS The volume of newly formed mineralized tissue in the leptin group was significantly (P < .01) higher than that in the control group based on micro-computed tomographic analysis. In histologic examination, hard tissue formation was rarely shown in the no pulp capping and Col + PBS groups. However, significantly (P < .01) larger amounts of newly mineralized tissue deposition were observed in the leptin groups. In immunohistochemical analysis, reparative dentin and new vessels formed in the pulp cavity of the leptin groups. Vascular endothelial growth factor, dentin sialoprotein, and dentin sialophosphoprotein were expressed around the newly formed mineralized tissue area. CONCLUSIONS Leptin showed the ability to induce angiogenesis, odontogenic differentiation, and mineralization in exposed rat pulps. Leptin also exhibited favorable inflammatory responses in the pulp tissue. Not only osteodentin but also tubular dentin and new vessels were observed in the pulp cavity.

Introduction. Elastin-like polypeptide (ELP) supplementation was previously reported to enhance the physical properties of mineral trioxide aggregate (MTA). The aim of this study was to investigate the effect of ELP supplementation on the bonding properties of MTA to dentin. Methods. Two types of ELPs were synthesized and mixed with MTA in a 0.3 liquid/powder ratio. The push-out bond strength test and interfacial observation with scanning electron microscopy were performed for ELP-supplemented MTA. The porosity of MTA fillings in the cavity was observed with microcomputed tomography. The stickiness, flow rate, and contact angle were additionally measured for potential increased bonding properties. Results. ELP supplementation improved the bond strength of MTA to dentin. MTA supplemented by a specific ELP exhibited a less porous structure, higher stickiness, and higher flow rate. ELPs also decreased the contact angle to dentin. Conclusions. This research data verifies that ELP improves the bonding properties of MTA to a tooth structure. The sticky and highly flowable characteristics of ELP-supplemented MTA may provide intimate contact with dentin and supply a less porous cement structure, which might improve the bonding properties of MTA.

The aim of this study was to compare chemical properties and bioactivities of with mineral trioxide aggregate (MTA) and EndoSequence Root Repair Material (ERRM). After setting, surfaces of test materials were observed by scanning electron microscope (SEM). The pH and cell viability of materials were tested. Osteoblastic differentiation and alkaline phosphatase (ALP) activity were measured by quantitative real-time PCR and ALP staining. MTA showed spindle-shaped crystals while ERRM showed round-shaped crystals of various sizes. ERRM presented lower pH than MTA. Both materials showed good cell viabilities compared to the control. Expression levels of osteoblastic genes and ALP staining were increased significantly (p

An ideal root canal sealer creates a bacteria-resistant seal and exhibits antimicrobial activity, biocompatibility, and osteoconductivity. The aim of this study was to assess the effects of 3 root canal sealers on cell viability, inflammatory response, and osteogenic potential in MC3T3-E1 cells.AH Plus (Dentsply Caulk, Milford, DE), MTA Fillapex (Angelus Solucxoes Odontologicas, Londrina, Brazil), and EndoSequence BC (Brasseler, Savannah, GA) were mixed according to the manufacturer's instructions, and samples were prepared as extraction media (final dilution: 1/10). Lipopolysaccharide (LPS) (100 ng/mL) treatment was used to induce an inflammatory response in this study. Cell viability was evaluated using the Water soluble tetrazolium-1 (WST-1) assay. The levels of inflammatory mediators (interleukin 6 and tumor necrosis factor alpha) and osteogenic marker genes (ALP and OCN) were measured by reverse-transcription polymerase chain reaction and real-time polymerase chain reaction. Osteogenic potential was evaluated by alkaline phosphatase staining and alizarin red staining.Calcium silicate-based sealers such as MTA Fillapex and EndoSequence BC showed strong cell viability compared with AH Plus. AH Plus, MTA Fillapex, and EndoSequence BC decreased the levels of LPS-induced inflammatory mediators (P .05). The expression of osteogenic marker genes, alkaline phosphatase activity, and mineralized nodule formation decreased with LPS treatment. However, AH Plus and calcium silicate-based sealers increased the osteogenic potential reduced by LPS treatment (P .05).Calcium silicate-based sealers exhibit anti-inflammatory effects and induce osteogenic differentiation in MC3T3-E1 cells.

This study compared the characteristics of periodontal ligament stem cells (PDLSCs) cultured using 3-dimensional (3D) versus conventional 2-dimensional (2D) methods. PDLSCs were cultured in either a 3D culture with a non-adhesive culture plate (Stemfit 3D®) or a conventional 2D culture using a 6-well plate. Morphology, viability, proliferation ability, and osteogenic differentiation were analyzed to characterize the differences induced in identical PDLSCs by 3D and 2D culture environments. In addition, gene expression was analyzed using RNA sequencing to further characterize the functional differences. The diameter and the viability of the 3D-cultured PDLSCs decreased over time, but the shape of the spheroid was maintained for 20 days. Although osteogenic differentiation occurred in both the 2D- and 3D-cultured PDLSCs, compared to the control group it was 20.8 and 1.6 higher in the 3D- and 2D-cultured cells, respectively. RNA sequencing revealed that PDLSCs cultured using 2D and 3D methods have different gene expression profiles. The viability of the 3D-cultured cells was decreased, but they showed superior osteogenic differentiation compared to 2D-cultured cells. Within the limitations of this study, the results demonstrate that the structure and function of PDLSCs are influenced by the cell culture method.

Although mineral trioxide aggregates (MTA) have been adopted as an endodontic sealer because of excellent sealing effect and bioactive property and been modified with improvement of its characteristics, the developed MTA sealers have not yet satisfied all the ideal requirements of endodontic sealers. The aim of this study was to assess the characteristics of elastin-like polypeptide (ELP)-incorporated MTA for use as an endodontic sealer and compare them with those of commercial MTA sealers. Two commercial MTA sealers and three experimental ELP-incorporated MTA sealers with 0.3, 0.4, and 0.5 liquid/powder (L/P) ratio for 10 wt% ELP liquid were evaluated. The push-out bond strength, flow rate, sealer penetrability and wash-out resistance were tested and the sealer-dentin interface was observed using a scanning electron microscope (SEM). Our study revealed the ELP-incorporated MTA sealer, especially in 0.4 L/P ratio, exhibited the higher push-out bond strength and flow rate (P

Root canal treatment of mandibular incisor is difficult because of the narrow pulp space and apical curvature. The aim of this study was to measure the anatomical indicators of the mandibular incisors in Koreans using micro-computed tomography (MCT) with novel software (Kappa 2). The MCT-scanned data from 27 mandibular incisors were reconstructed and analyzed. For each canal, 3-dimensional (3D) surface models were re-sliced at 0.1 mm intervals perpendicular to the central axis of the root canal. Root canal width, dentine thickness, and direction and degree of root canal curvatures were measured automatically on each slice. Measurements were analyzed statistically with Bhapkar test, Friedman test, and Wilcoxon signed rank test. Labial and lingual dentine thicknesses were significantly larger than mesial and distal thicknesses (p &lt, 0.001). The thinnest dentine was mainly located on the mesio-lingual side of the canals in the apical third. The mean narrowest and widest canal width in the apical sixth were 0.22 mm and 0.40 mm, respectively. The canal curvature abruptly increased in the apical 0.5-mm portion. MCT with novel software provided useful anatomical information for root canal instrumentation.

Various three-dimensional (3D) culture methods have been introduced to overcome the limitations of in vitro culture and mimic in vivo conditions. This study aimed to evaluate two microsphere-forming culture methods and a monolayer culture method. We evaluated cell morphology, viability, osteo-, adipo-, and chondrogenic differentiation potential of dental pulp stem cells (DPSCs) cultured in 3D culture plates: ultra-low attachment (ULA) and U-bottomed StemFit 3D (SF) plates, and a two-dimensional (2D) monolayer plate. RNA sequencing (RNA-seq) revealed differentially expressed gene (DEG) profiles of the DPSCs. In contrast to an increasing pattern in the 2D group, cell viability in 3D groups (ULA and SF) showed a decreasing pattern, however, high multilineage differentiation was observed in both the 3D groups. RNA-seq showed significantly overexpressed gene ontology categories including angiogenesis, cell migration, differentiation, and proliferation in the 3D groups. Hierarchical clustering analysis revealed a similar DEG regulation pattern between the 3D groups, however, a comparatively different DEG was observed between the 2D and 3D groups. Taken together, this study shows that DPSCs cultured in microsphere-forming plates present superior multilineage differentiation capacities and demonstrate higher DEG expression in regeneration-related gene categories compared to that in DPSCs cultured in a conventional monolayer plate.

Introduction Few studies have reported direct pulp capping in inflamed pulp conditions. The purpose of this study was to investigate the in vitro and in vivo responses of dental pulp during direct pulp capping using various pulp capping materials in inflamed conditions. Methods Human dental pulp cells were treated with lipopolysaccharide (LPS) and cultured with Dycal (Dentsply Caulk, Milford, DE), ProRoot MTA (Dentsply Maillefer, Ballaigues, Switzerland), and Endocem MTA (Maruchi, Wonju, South Korea). The expressions of interleukin (IL)-1β, IL-6, dentin matrix protein 1, and dentin sialophosphoprotein were analyzed through real-time polymerase chain reaction. The maxillary molars of Sprague-Dawley rats were exposed for 2 days. The exposed pulps were capped with Dycal, ProRoot MTA, and Endocem MTA and sealed with resin-modified glass ionomer followed by histologic and immunohistochemical analyses. Results The expression of IL-1β and IL-6 was increased with LPS and decreased by Dycal, ProRoot MTA, and Endocem MTA. Dentin matrix protein 1 and dentin sialophosphoprotein levels were decreased with LPS and increased after treatment with pulp capping materials.In the in vivo study, inflammation associated with Dycal was higher than that associated with ProRoot MTA and Endocem MTA at week 1, without any significant difference between the 2. At 4 weeks, inflammation was decreased, and mineralization was increased compared with week 1 in all 3 of the materials. At week 1, IL-6 immunoreactivity was strongly expressed. Dycal exhibited stronger immunoreactivity than ProRoot MTA and Endocem MTA. However, the immunoreactivity was decreased in all groups at week 4. Conclusions Successful direct pulp capping requires more effective pulp capping materials for the treatment of inflamed pulps.

Premature surgical menopause (PSM) without subsequent hormone replacement therapy (HRT) can lead to morbidity and mortality. Our objective was to describe the use of HRT following PSM and identify variables associated with HRT use based on prescription records from a population-based cohort.A population-based retrospective cohort of women in British Columbia, Canada who underwent PSM between the ages of 19 and 49 years. Women were identified using surgical data from the Discharge Abstract Database and linked to HRT prescription histories from the BC PharmaNet database for the period of 2004 to 2014. HRT prescription rates were calculated, and factors associated with postoperative HRT use were identified.A total of 12 837 women were included, with a median age of 43 years. They had undergone BSO with concurrent hysterectomy (49.9%). bilateral salpingo-oophorectomy (BSO) alone (42.1%), or bilateral oophorectomy (BO) (8%). The most common indications for surgery were endometriosis (17.9%), benign adnexal neoplasm (17.2%), and abnormal bleeding (14.0%). Only 55.3% of women ever used HRT, and 47.9% of these women used HRT for less than 1 year. HRT use was higher among women who underwent concurrent hysterectomy (60.7% vs. 50%, P0.001). This association remained significant after multivariate adjustment (aOR 1.64; 95% CI 1.50-1.79). Women with a known BRCA mutation were also more likely to use HRT postoperatively (aOR 3.73; 95% CI 2.14-6.81).In this large population-based study, HRT use after PSM was 50%. Our study highlights the need for education of both health care providers and patients, and for ongoing follow-up in this young population.

Background/purpose: Matrix metalloproteinases (MMPs) play a crucial role in the pathogenesis of dental caries, collapse of adhesive interface, and chemical erosion of teeth. The objective of this study was to investigate the inhibitory effect of zinc on collagen degradation. Materials and methods: Human dentin was ground and demineralized by citric acid (pH 2.0). The demineralized ground dentin was incubated in six different media: artificial saliva (AS); 5 mg/ml doxycycline in AS; 3.33, 6.82, 13.63, and 27.26 mg/ml of zinc chloride (Zn) in AS. Each group was divided into two subgroups, and active MMP-2 was incorporated into one subgroup. Specimens were incubated for 24 h, 1 week, and 2 weeks. Collagen degradation product was assessed using ELISA. The results were analyzed using repeated measured ANOVA and Duncan's post hoc analysis (α = 0.05). Results: The amount of collagen degradation was the lowest in Doxy group. Zn groups showed a significant inhibitory effect in collagen degradation for all concentrations (P

Purpose This study aims to compare the volumetric change, degree of conversion (DOC), and cytotoxicity of 3D-printed restorations post-cured under three different conditions. Materials and methods 3D-printed interim restorations were post-cured under three different conditions and systems: 5 min, 30 min, and 24 h. Three-unit and six-unit fixed dental prostheses (n = 30 for each case) were printed; ten specimens from each group were post-cured and then scanned to compare their volumetric changes. Root-mean-squared (RMS) values of the data were acquired by superimposing the scanned files with original files. Thirty disk-shaped specimens were printed to evaluate the DOC ratio. Fourier transform infrared spectroscopy was used to compare the DOCs of 10 specimens from each group. Human gingival fibroblasts were used to measure the cell viability of every specimen (n = 7). The data from this experiment were employed for one-way analysis of variance and Tukey's post-hoc comparisons. Results Differences between the three-unit restorations were statistically insignificant, regardless of the post-curing conditions. However, for the six-unit restorations, a high RMS value was acquired when the post-curing duration was 30 min. The average DOC was approximately 56 - 62%; the difference between each group was statistically insignificant. All the groups exhibited cell viability greater than 70%, rendering them clinically acceptable. Conclusion The post-curing conditions influenced the volume when the length of the restoration was increased. However, this deviation was found to be clinically acceptable. Additionally, post-curing did not significantly influence the DOC and cytotoxicity of the restorations.

Uterine serosal pregnancy is an extremely rare form of ectopic pregnancy. This is a report of a 35-year-old primigravida woman who was diagnosed with uterine serosal pregnancy via laparoscopic intervention. A 35-year-old woman (gravida 1, para 0) was referred from a local clinic for a ruptured left tubal pregnancy at amenorrhea 5+0 weeks with elevated serum beta human chorionic gonadotropin (16,618 mIU/mL). A pregnancy on the left posterior wall of the uterine serosa was diagnosed during the operation and successfully treated with laparoscopic surgery as a conservative management strategy to enable fertility preservation. With the advantages of ultrasonography and laparoscopy, an early diagnosis of a primary abdominal pregnancy located on the left posterior wall of the uterine serosa was made, prior to the occurrence of severe intra-abdominal massive hemorrhage, which was then treated laparoscopically as a conservative management strategy enabling the preservation of fertility.

SUMMARY The aim of this randomized controlled clinical trial was to compare the clinical effectiveness of different polishing systems and self-etch adhesives in class V composite resin restorations. A total of 164 noncarious cervical lesions (NCCLs) from 35 patients were randomly allocated to one of four experimental groups, each of which used a combination of polishing systems and adhesives. The two polishing systems used were Sof-Lex XT (Sof), a multistep abrasive disc, and Enhance/Pogo (EP), a simplified abrasive-impregnated rubber instrument. The adhesive systems were Clearfil SE bond (CS), a two-step self-etch adhesive, and Xeno V (XE), a one-step self-etch adhesive. All NCCLs were restored with light-cured microhybrid resin composites (Z250). Restorations were evaluated at baseline and at 6, 12, 18, and 24 months by two blinded independent examiners using modified FDI criteria. The Fisher exact test and generalized estimating equation analysis considering repeated measurements were performed to compare the outcomes between the polishing systems and adhesives. Three restorations were dislodged: two in CS/Sof and one in CS/EP. None of the restorations required any repair or retreatment except those showing retention loss. Sof was superior to EP with regard to surface luster, staining, and marginal adaptation (p0.05). Sof is clinically superior to EP for polishing performance in class V composite resin restoration. XE demonstrates clinically equivalent bonding performance to CS.

Objectives To aid primary care physicians, emergency medicine physicians, and gynaecologists in the initial investigation of adnexal masses, defined as lumps that appear near the uterus or in or around ovaries, fallopian tubes, or surrounding connective tissue, and to outline recommendations for identifying women who would benefit from a referral to a gynaecologic oncologist for further management. Intended Users Gynaecologists, obstetricians, family physicians, general surgeons, emergency medicine specialists, radiologists, sonographers, nurses, medical learners, residents, and fellows. Target Population Adult women 18 years of age and older presenting for the evaluation of an adnexal mass. Options Women with adnexal masses should be assessed for personal risk factors, history, and physical findings. Initial evaluation should also include imaging and laboratory testing to triage women for management of their care either by a gynaecologic oncologist or as per SOGC guideline no. 404 on the initial investigation and management of benign ovarian masses. Evidence A search of PubMed, Cochrane Wiley, and the Cochrane systematic reviews was conducted in January 2018 for English-language materials involving human subjects published since 2000 using three sets of terms: (i) ovarian cancer, ovarian carcinoma, adnexal disease, ovarian neoplasm, adnexal mass, fallopian tube disease, fallopian tube neoplasm, ovarian cyst, and ovarian tumour; (ii) the above terms in combination with predict neoplasm staging, follow-up, and staging; and (iii) the above two sets of terms in combination with ultrasound, tumour marker, CA 125, CEA, CA19-9, HE4, multivariable-index-assay, risk-of-ovarian-malignancy-algorithm, risk-of-malignancy-index, diagnostic imaging, CT, MRI, and PET. Relevant evidence was selected for inclusion in descending order of quality of evidence as follows: meta-analyses, systematic reviews, guidelines, randomized controlled trials, prospective cohort studies, observational studies, non-systematic reviews, case series, and reports. Additional articles were identified through cross-referencing the identified reviews. The total number of studies identified was 2350, with 59 being included in this review. Validation Methods The content and recommendations were drafted and agreed upon by the authors. The Executive and Board of the Society of Gynecologic Oncology of Canada reviewed the content and submitted comments for consideration. The Board of Directors of the Society of Obstetricians and Gynaecologists of Canada approved the final draft for publication. The quality of evidence was rated using the criteria described in the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) methodology framework (Table A1 of Online Appendix A). See Table A2 of Online Appendix A for the interpretation of strong and weak recommendations. The summary of findings is available upon request. Benefits, Harms, Costs Adnexal masses are common, and guidelines on how to triage them and manage the care of patients presenting with adnexal masses will continue to guide the practice of primary care providers and gynaecologists. Ovarian cancer outcomes are improved when initial surgery is performed by a gynaecologic oncologist, likely as a result of complete surgical staging and optimal cytoreduction. Given these superior outcomes, guidelines to assist in the triage of adnexal masses and the referral and management of the care of patients with an adnexal mass are critical. SUMMARY STATEMENTS (GRADE ratings in parentheses) 1Information from a detailed personal and family history can guide decisions on further testing and evaluation for women with adnexal masses (low). 2The majority of women with ovarian cancer have symptoms in the year prior to their diagnosis, although these symptoms can be vague (low). 3Adnexal masses are best evaluated through subjective pattern recognition by an expert sonographer in order to distinguish benign adnexal masses from malignant adnexal masses (moderate). 4Ultrasound evaluation of adnexal masses with risk prediction models such as the simple rules, developed by the International Ovarian Tumor Analysis group, allows sonographers with varying degrees of expertise to use uniform terminology and accurately classify these masses as "likely malignant," "benign," or "indeterminate" (moderate). 5Laboratory testing can aid in the differential diagnosis of an adnexal mass. Testing for sexually transmitted infection with results showing leukocytosis can help identify tubo-ovarian abscesses, while a positive pregnancy test can lead to diagnosis of possible ectopic pregnancies (low). 6Cancer antigen 125 is a non-specific glycoprotein that can be elevated in benign and malignant gynaecologic conditions and in non-gynaecologic conditions (moderate). 7Only half of all early stage ovarian cancers and 80% of advanced stage ovarian cancers show elevated cancer antigen 125 levels (moderate). 8The sensitivity and specificity of cancer antigen 125 as a tumour marker are higher in women who are postmenopausal because many of the benign clinical conditions that can increase the level of cancer antigen 125 occur in the premenopausal population, while most cases of epithelial ovarian, fallopian tube, and peritoneal cancer occur in the postmenopausal population (moderate). 9Despite having gone through several iterations, the risk of malignancy index, which uses cancer antigen 125 and sonographic features along with the patient's menopausal status, is outperformed by the more recent International Ovarian Tumor Analysis group's logistic regression model 2 and simple rules. The sensitivity and specificity of version 2 of the risk of malignancy index for distinguishing malignant from benign masses are only 75% and 87%, respectively (moderate). RECOMMENDATIONS (GRADE ratings in parentheses) 1Take a detailed history for a woman presenting with an adnexal mass. For a woman with a personal history of infertility, endometriosis, or cancer or a family history of cancer, refer the patient to a gynaecologic oncologist for further evaluation if possible (strong, low). 2Physical examination should include lymph node survey, respiratory examination to rule out pleural effusion or consolidation, breast and axillary examination to rule out breast malignancy, and an abdominal examination to assess for ascites, omental caking, and organomegaly as well as a pelvic examination, including a bimanual and rectovaginal examination, to assess for mass size, contour, mobility, and parametrial, bladder, and rectal abnormality (strong, low). 3If a woman presents with an adnexal mass, request an initial ultrasound using either pattern recognition or the risk prediction model developed by the International Ovarian Tumour Analysis group (strong, moderate). 4Promptly refer to a gynaecologic oncologist any patient who presents with a mass with any of the following sonographic features, suggestive of malignancy: (i) solid component with strong or central colour flow, (ii) ≥4 papillary projections (defined as >3 mm in height), (iii) thick multiple irregular septations, or (iv) ascites and peritoneal nodularity. While awaiting a gynaecologic oncologist consult, where resources permit, pursue further investigations, including tumour marker levels and computed tomography scan of the chest, abdomen, and pelvis, as appropriate (strong, moderate). 5Women with adnexal masses with indeterminate features should be evaluated by ultrasound conducted by an expert sonographer (if available) or by magnetic resonance imaging or be referred to a gynaecologic oncologist (strong, low). 6Do not use cancer antigen 125 testing as a screening tool in asymptomatic women without a pelvic or adnexal mass (strong, moderate). 7The initial ultrasound characterization can be done using either pattern recognition (best in the hands of an expert sonographer) or risk prediction algorithms, such as simple rules, which have been demonstrated to work well for practitioners with varying degrees of expertise. If the lesion is suspicious for malignancy, prompt referral to a gynaecologic oncologist is recommended (strong, moderate). 8In women 9Further tumour marker testing, including carcinoembryonic antigen, cancer antigen 19-9, and cancer antigen 15-3, along with referral to a gynaecologic oncologist are recommended for women presenting with bilateral masses with features of malignancy (strong, moderate).

Nickel-titanium (NiTi) rotary files have enabled efficient root canal preparations that maintain the canal center with fewer aberrations compared to hand files. However, NiTi rotary files are susceptible to fracture, which can thereby compromise root canal treatment. Therefore, NiTi files have been developed to enhance fracture resistance by modifying design and thermal treatment. The objective of this study was to compare the torsional fatigue resistance and bending resistance of NiTi files manufactured from different alloys and treatments. ProTaper NEXT X2 (PTN; M-wire), V taper 2H (V2H; controlled memory wire), NRT (heat-treated), and One Shape (OS; conventional alloy) instruments of tip size #25 were compared. Torsional fatigue was evaluated by embedding the 3 mm tip of each instrument (N = 10/brand) in resin and the repetitive application of torsional stress (300 rpm, 1.0 N·cm) by an endodontic motor with autostop when the file fractured. The number of loading cycles to fracture was recorded and analyzed by Kruskal–Wallis and Mann–Whitney U tests with Bonferroni’s correction. Bending resistance of the instruments was tested using a cantilever bending test to the 3 mm point from the tip (N = 10/brand). The stress was measured when deflection of 3 mm was subjected and statistically analyzed with a one-way analysis of variance and Tukey’s honest significance difference test (α = 0.05). V2H withstood the highest number of load applications during torsional fatigue testing (p<0.05), followed by NRT, PTN, and OS, where the differences between NRT and PTN (p=0.035) and between PTN and OS (p=0.143) were not statistically significant. V2H showed the lowest bending stiffness, followed by NRT, PTN, and OS (p<0.001). Thermal treatment of NiTi wire resulted in improved mechanical properties, and controlled memory wire provided improved flexibility and torsional fatigue resistance.

Angiotensin II (AngII) triggers a transient contraction of pulmonary arteries (PAs) followed by protracted desensitization. Based on the unconventional eNOS expression in PA smooth muscle cells (PASMCs), we hypothesized that activation of smooth muscle eNOS by AngII might be responsible for fast relaxation and tachyphylaxis. Using dual-wire myograph, mechanically endothelium-denuded rat PA [E(−)PA] showed AngII concentration–dependent transient contractions (ΔTAngII, 95% decay within 1 min), which were abolished by losartan (AT1R antagonist). Neither PD123319 (AT2R antagonist) nor A779 (MasR antagonist) affected ΔTAngII. When the vessels were pretreated with L-NAME (NOS inhibitor), ODQ (guanylate cyclase inhibitor), or KT5823 (PKG inhibitor), ΔTAngII of E(−)PA became larger and sustained, whereas nNOS or iNOS inhibitors had no such effect. Immunoblotting of human PASMCs (hPASMCs) also showed eNOS expression, and AngII treatment induced activating phosphorylations of Ser1177 in eNOS and of Ser473 in Akt (Ser/Thr protein kinase B), an upstream signal of eNOS phosphorylation. In addition, L-NAME co-treatment promoted AngII-induced Ser19 phosphorylation of myosin light chain. In hPASMCs, AngII abolished plasma membrane expression of AT1R, and recovery by washout took more than 1 h. Consistent with the data from hPASMCs, the second application of AngII to E(−)PA did not induce contraction, and significant recovery of ΔTAngII required prolonged washout (> 2 h) in the myography study. L-NAME treatment before the second application facilitated recovery of ΔTAngII. Muscular eNOS plays an auto-inhibitory role in ΔTAngII of PAs. The molecular changes investigated in hPASMCs revealed eNOS phosphorylation and internalization of AT1R by AngII. We propose that the rat PA smooth muscle eNOS-induced lusitropy and slow recovery of AT1R from tachyphylaxis might counterbalance the excessive contractile response to AngII, contributing to the distinctive low-pressure pulmonary circulation.

The purpose of this study is to assess the effect of autoclave sterilization on the cyclic fatigue and torsional fracture resistance of ProTaper Universal (PTU), K3XF, HyFlex EDM (EDM), and TF adaptive (TFA). Sixty instruments from each file type were divided into two categories for cyclic fatigue group (CGr) and torsional fracture group (TGr). CGr and TGr were divided into three subgroups, respectively, consisting of ten instruments from each file type. Cyclic fatigue fracture test was performed using artificial canal made of stainless steel, and the mean number of cycles to failure (NCF) were determined. CGr1, the files were tested to establish baseline for NCF; CGr2, the files were tested cyclic fatigue after 10 cycles of autoclave; CGr3, instruments were autoclaved after being cycled to 25, 50, and 75% of corresponding NCF determined in CGr1, followed by cyclic fatigue test. The torsional fracture test was performed without autoclave (TGr1), after 3-cycle autoclave (TGr2), and 7-cycle autoclave (TGr3), respectively, which evaluated maximum torque and angular deflection. NCF, maximum torque and angular deflection were compared using one-way ANOVA with Bonferroni test. Two-way ANOVA was performed to determine the interaction between 'autoclave treatment' and 'type of NiTi file'. EDM showed highest NCF within the same autoclave treatment. TFA presented the lowest maximum torque and the highest angular deflection, and PTU presented the lowest angular deflection. Within the same NiTi file systems, most of NCF, maximum torque and angular deflection of tested files were not significantly influenced by autoclave condition.

Mucopolysaccharidosis (MPS) is an inherited metabolic disorder caused by a deficiency in enzymes that participate in the degradation of glycosaminoglycans (GAGs) such as heparin sulfate and dermatan sulfate. Left untreated, patients show progressive mental and physical deterioration due to deposition of GAGs in organs. Death often occurs due to cardiac or respiratory failure before patients reach their early twenties. MPS has several oral and dental manifestations. An enlarged head, short neck, and open mouth associated with a large tongue are major characteristics of MPS patients. Dental complications can be severe, including unerupted dentition, dentigerous cyst-like follicles, malocclusions, condylar defects, and gingival hyperplasia. A 21-year-old female patient with MPS was described in this article, with special emphasis on oral manifestations and dental treatment.

Objectif Cette directive passe en revue l'evaluation clinique et la prise en charge des maladies gestationnelles trophoblastiques, notamment les traitements chirurgicaux et medicamenteux des tumeurs benignes, premalignes et malignes. L'objectif de la presente directive clinique est d'aider les fournisseurs de soins de sante a rapidement diagnostiquer les maladies gestationnelles trophoblastiques, a normaliser les traitements et le suivi et a assurer des soins specialises precoces aux patientes dont l'atteinte est maligne ou metastatique. PROFESSIONNELS CONCERNeS: Gynecologues generalistes, obstetriciens, medecins de famille, sages-femmes, urgentologues, anesthesistes, radiologistes, anatomopathologistes, infirmieres autorisees, infirmieres praticiennes, residents, gynecologues-oncologues, oncologues medicaux, radio-oncologues, chirurgiens, omnipraticiens en oncologie, infirmieres en oncologie, pharmaciens, auxiliaires medicaux et autres professionnels de la sante qui traitent des patientes atteintes d'une maladie gestationnelle trophoblastique. La presente directive vise egalement a fournir des renseignements aux parties interessees qui prodiguent des soins de suivi a ces patientes apres le traitement. Population cible Femmes en âge de procreer atteintes d'une maladie gestationnelle trophoblastique. Options Les femmes ayant recu un diagnostic de maladie gestationnelle trophoblastique doivent etre orientees vers un gynecologue afin qu'il realise une evaluation initiale, envisage une intervention chirurgicale primaire (evacuation ou hysterectomie) et effectue un suivi. Il y a lieu d'orienter les femmes ayant recu un diagnostic de tumeur trophoblastique gestationnelle vers un gynecologue-oncologue afin qu'il effectue la stadification tumorale, etablisse le score de risque et envisage l'intervention chirurgicale primaire ou un traitement systemique (mono- ou polychimiotherapie) et la necessite d'eventuels traitements supplementaires. Il est recommande de discuter de chaque cas de neoplasie gestationnelle trophoblastique lors d'une reunion multidisciplinaire de cas oncologiques et de l'inscrire dans une base de donnees centralisee (regionale et/ou nationale). DONNeES PROBANTES: Des recherches ont ete effectuees au moyen des bases de donnees Embase et MEDLINE, du Cochrane Central Register of Controlled Trials et de la Cochrane Database of Systematic Reviews afin de trouver les etudes publiees depuis 2002 utilisant un ou plusieurs des mots cles suivants : trophoblastic neoplasms, choriocarcinoma, trophoblastic tumor, placental site, gestational trophoblastic disease, hydatidiform mole, drug therapy, surgical therapy, radiotherapy, cure, complications, recurrence, survival, prognosis, pregnancy outcome, disease outcome, treatment outcome et remission. La recherche initiale a ete effectuee en avril 2017; une mise a jour a ete faite en mai 2019. Les donnees probantes pertinentes ont ete selectionnees aux fins d'inclusion selon l'ordre suivant : meta-analyses, revues systematiques, directives cliniques, essais cliniques randomises, etudes de cohortes prospectives, etudes observationnelles, revues non systematiques, etudes de series de cas et rapports. D'autres articles pertinents ont ete trouves en recoupant les revues repertoriees. Le nombre total d'etudes relevees etait de 673, dont 79 etudes sont citees dans la presente revue. MeTHODES DE VALIDATION: Le contenu et les recommandations ont ete rediges et acceptes par les auteurs. La direction et le conseil d'administration de la Societe de gyneco-oncologie du Canada ont passe en revue le contenu de la version preliminaire et ont soumis des commentaires a prendre en consideration. Le conseil d'administration de la Societe des obstetriciens et gynecologues du Canada a approuve la version definitive aux fins de publication. La qualite des donnees probantes a ete evaluee au moyen des criteres de l'approche GRADE (Grading of Recommendations Assessment, Development and Evaluation). Consulter les tableaux dans l'annexe en ligne pour connaitre les criteres de notation et d'interpretation des recommandations. BeNeFICES, RISQUES, COuTS: Les presentes recommandations aideront les medecins a diagnostiquer rapidement les maladies gestationnelles trophoblastiques et a orienter de facon urgente les patientes ayant recu un diagnostic de maladie gestationnelle trophoblastique en gynecologie oncologique pour une prise en charge specialisee. Le traitement des neoplasies gestationnelles trophoblastiques en centre specialise combine a l'utilisation de bases de donnees centralisees permet de recueillir et de comparer des donnees sur les resultats therapeutiques des patientes atteintes de ces tumeurs rares et d'optimiser les soins aux patientes. DECLARATIONS SOMMAIRES (CLASSEMENT GRADE ENTRE PARENTHeSES): RECOMMANDATIONS (CLASSEMENT GRADE ENTRE PARENTHeSES).

Objective This guideline reviews the clinical evaluation and management of gestational trophoblastic diseases, including surgical and medical management of benign, premalignant, and malignant entities. The objective of this guideline is to assist health care providers in promptly diagnosing gestational trophoblastic diseases, to standardize treatment and follow-up, and to ensure early specialized care of patients with malignant or metastatic disease. Intended users General gynaecologists, obstetricians, family physicians, midwives, emergency department physicians, anaesthesiologists, radiologists, pathologists, registered nurses, nurse practitioners, residents, gynaecologic oncologists, medical oncologists, radiation oncologists, surgeons, general practitioners in oncology, oncology nurses, pharmacists, physician assistants, and other health care providers who treat patients with gestational trophoblastic diseases. This guideline is also intended to provide information for interested parties who provide follow-up care for these patients following treatment. Target population Women of reproductive age with gestational trophoblastic diseases. Options Women diagnosed with a gestational trophoblastic disease should be referred to a gynaecologist for initial evaluation and consideration for primary surgery (uterine evacuation or hysterectomy) and follow-up. Women diagnosed with gestational trophoblastic neoplasia should be referred to a gynaecologic oncologist for staging, risk scoring, and consideration for primary surgery or systemic therapy (single- or multi-agent chemotherapy) with the potential need for additional therapies. All cases of gestational trophoblastic neoplasia should be discussed at a multidisciplinary cancer case conference and registered in a centralized (regional and/or national) database. Evidence Relevant studies from 2002 onwards were searched in Embase, MEDLINE, the Cochrane Central Register of Controlled Trials, and Cochrane Systematic Reviews using the following terms, either alone or in combination: trophoblastic neoplasms, choriocarcinoma, trophoblastic tumor, placental site, gestational trophoblastic disease, hydatidiform mole, drug therapy, surgical therapy, radiotherapy, cure, complications, recurrence, survival, prognosis, pregnancy outcome, disease outcome, treatment outcome, and remission. The initial search was performed in April 2017 and updated in May 2019. Relevant evidence was selected for inclusion in the following order: meta-analyses, systematic reviews, guidelines, randomized controlled trials, prospective cohort studies, observational studies, non-systematic reviews, case series, and reports. Additional significant articles were identified through cross-referencing the identified reviews. The total number of studies identified was 673, with 79 studies cited in this review. Validation methods The content and recommendations were drafted and agreed upon by the authors. The Executive and Board of Directors of the Society of Gynecologic Oncology of Canada reviewed the content and submitted comments for consideration, and the Board of Directors for the Society of Obstetricians and Gynaecologists of Canada approved the final draft for publication. The quality of evidence was rated using the criteria described in the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) methodology framework. See the online appendix tables for key to grading and interpretation of recommendations. Benefits These guidelines will assist physicians in promptly diagnosing gestational trophoblastic diseases and urgently referring patients diagnosed with gestational trophoblastic neoplasia to gynaecologic oncology for specialized management. Treating gestational trophoblastic neoplasia in specialized centres with the use of centralized databases allows for capturing and comparing data on treatment outcomes of patients with these rare tumours and for optimizing patient care. Summary statements (grade ratings in parentheses) RECOMMENDATIONS (GRADE RATINGS IN PARENTHESES).

Objectives This randomized double-blinded clinical trial evaluated the bleaching efficacy and incidence of contact hypersensitivity of three kinds of bleaching toothpaste. Methods Forty-nine participants above A2 shade on the maxillary central incisor (#11) and canine (#13) were randomized into three groups: TW group (n = 15), 0.75 % HP-containing toothpaste (Toothwhole white); VL group (n = 15), 0.75 % HP-containing toothpaste (Vussen 7); and VH group (n = 17), 2.8 % of HP-containing toothpaste (Vussen 28). Participants were instructed to manually brush their teeth for 3 min, 3 times per day for 12 weeks. They were followed-up after 4 and 12 weeks. Shade measurements were performed using a spectrophotometer (SP), and data were calculated with CIELab ( Δ E a b * ) and CIEDE2000 ( Δ E 00 ) formula. Additionally, visual inspection (VI) using the Vitapan classical shade guide was also performed, and the correlation between the two measurements was analyzed by comparing the CIELab ( Δ E a b * ) values. The incidence of contact hypersensitivity at each follow-up was recorded. A mixed-effect model was performed to assess shade changes and chi-square tests for the incidence of contact hypersensitivity, respectively. Results At 12 week follow-up, all groups showed Δ E a b * above 3.46, and Δ E 00 above 2.25 for tooth #11. For tooth #13, the Δ E a b * and Δ E 00 of VH and TW groups only were above those thresholds. Shade change varied according to toothpaste and follow-up points. SP and VI showed a moderate positive correlation for L*, a*, and b*, respectively (P 0.05). Conclusions Bleaching toothpaste with higher HP yields a better shade change than other toothpaste after 12 weeks. Clinical significance The use of bleaching toothpaste with a higher HP concentration results in a better shade improvement.

Objectives In this study, we characterized human dental pulp cells (HDPCs) obtained by different culture methods to establish the most suitable methodology for dental tissue engineering and regenerative endodontic applications. Materials and Methods HDPCs were isolated by the outgrowth method (HDPCs-OG), the enzymatic digestion method (collagenase/dispase/trypsin, HDPCs-ED), or the combination of both methods (HDPCs-Combined). The expression of mesenchymal stem cell markers (CD105, CD90, and CD73) was investigated. In vitro differentiation capacities of HDPCs into adipogenic, osteogenic, and chondrogenic lineages were compared. Differentiation markers were analyzed by quantitative reverse-transcription polymerase chain reaction (RT-PCR) and western blotting. Results Our data indicated that whole HDPCs-ED, HPDCs-OG, and HDPCs-Combined could be differentiated into adipogenic, chrondrogenic, and osteogenic cell types. However, we found that the methods for isolating and culturing HDPCs influence the differentiation capacities of cells. HDPCs-OG and HDPCs-ED were preferably differentiated into adipogenic and osteogenic cells, respectively. Differentiation markers shown by RT-PCR and western blotting analysis were mostly upregulated in the treated groups compared with the control groups. Conclusions Our findings confirmed that cell populations formed by two different culture methods and the combined culture method exhibited different properties. The results of this study could provide an insight into regenerative endodontic treatment using HDPCs.

Pulmonary arteries (PAs) have high compliance, buffering the wide ranges of blood flow. Here, we addressed a hypothesis that PA smooth muscle cells (PASMCs) express nitric oxide synthases (NOS) that might be activated by mechanical stress and vasoactive agonists. In the myograph study of endothelium-denuded rat PAs, NOS inhibition (L-NAME) induced strong contraction (96 % of 80 mM KCl-induced contraction (80K)) in the presence of 5 nM U46619 (thromboxane A2 (TXA2) analogue) with relatively high basal stretch (2.94 mN, S(+)). With lower basal stretch (0.98 mN, S(-)), however, L-NAME application following U46619 (TXA2/L-NAME) induced weak contraction (27 % of 80K). Inhibitors of nNOS and iNOS had no such effect in S(+) PAs. In endothelium-denuded S(+) mesenteric and renal arteries, TXA2/L-NAME-induced contraction was only 18 and 21 % of 80K, respectively. Expression of endothelial-type NOS (eNOS) in rat PASMCs was confirmed by RT-PCR and immunohistochemistry. Even in S(-) PAs, pretreatment with H2O2 (0.1-10 μM) effectively increased the sensitivity to TXA2/L-NAME (105 % of 80K). Vice versa, NADPH oxidase inhibitors, reactive oxygen species scavengers, or an Akt inhibitor (SC-66) suppressed TXA2/L-NAME-induced contraction in S(+) PAs. In a human PASMC line, immunoblot analysis showed the following: (1) eNOS expression, (2) Ser(1177) phosphorylation by U46619 and H2O2, and (3) Akt activation (Ser(473) phosphorylation) by U46619. In the cell-attached patch clamp study, H2O2 facilitated membrane stretch-activated cation channels in rat PASMCs. Taken together, the muscular eNOS in PAs can be activated by TXA2 and mechanical stress via H2O2 and Akt-mediated signaling, which may counterbalance the contractile signals from TXA2 and mechanical stimuli.

This study evaluates the effects of various agitation methods on the adhesive layer formation of a new HEMA-free universal dentin adhesive. The µTBS of the universal adhesive, G-Premio BOND in the self-etch mode was evaluated using three agitation methods [passive agitation (PA), active agitation (AA), ultrasonic agitation (UA)], with and without aging treatment. Two-way analysis of variance revealed that aging treatment was not a statistically significant factor. Tukey's HSD test showed significant differences based on the application method, UA>AA>PA. TEM images of the PA group revealed multiple water blisters in the adhesive layers; AA and UA groups presented significantly less or no blisters within the adhesive layers; thus, AA and UA groups exhibited better bonding performance for the HEMA-free universal adhesive. It is assumed that the entrap ped blisters can be reduced with the active application of dentin adhesive, and thus improving the bonding performance.

This case report describes an innovative virtual simulation method using a computer-aided rapid prototyping (CARP) model and a computer-aided design (CAD) program for autotransplantation of an immature third molar.A compromised left mandibular second molar (#18) was extracted and replaced by autotransplantation using an immature left mandibular third molar (#17). In order to minimize the surgical time and injury to the donor tooth, a virtual 3-dimensional (3D) rehearsal surgery was planned. Cone-beam computed tomographic images were taken to fabricate the 3D printing CARP model of the donor tooth and tentative extraction socket. Subsequently, both CARP models were scanned with an intraoral scanner (CEREC Omnicam; Dentsply Sirona, Bensheim, Germany) followed by superimposition and virtual simulation of osteotomy preparation of the recipient alveolus using the CAD analysis program. During the surgery, the extraction socket was precisely prepared according to the predetermined location and dimensions via virtual simulation rehearsal surgery using CAD analysis. The donor tooth was atraumatically transplanted into the prepared socket. The follow-up examination revealed that the root developed with a normal periodontal ligament and lamina dura.Virtual simulation using a 3D printing CARP model and a CAD program could be clinically useful in autotransplantation of an immature third molar by ensuring an atraumatic and predictable surgery.

Objectives The purpose of this study was to evaluate the effect of bioactive glass (BAG)-containing composite on dentin remineralization. Methods Sixty-six dentin disks with 3 mm thickness were prepared from thirty-three bovine incisors. The following six experimental groups were prepared according to type of composite (control and experimental) and storage solutions (simulated body fluid [SBF] and phosphate-buffered saline [PBS]): 1 (undemineralized); 2 (demineralized); 3 (demineralized with control composite in SBF); 4 (demineralized with control composite in PBS); 5 (demineralized with experimental composite in SBF); and 6 (demineralized with experimental composite in PBS). BAG65S (65% Si, 31% Ca, and 4% P) was prepared via the sol-gel method. The control composite was made with a 50:50 Bis-GMA:TEGDMA resin matrix, 57 wt% strontium glass, and 15 wt% aerosol silica. The experimental composite had the same resin and filler, but with 15 wt% BAG65S replacing the aerosol silica. For groups 3–6, composite disks (20 × 10 × 2 mm) were prepared and approximated to the dentin disks and stored in PBS or SBF for 2 weeks. Micro-hardness test, attenuated total reflection Fourier-transform infrared spectroscopy (ATR-FTIR) and field-emission scanning electron microscopy (FE-SEM) was investigated. Results The BAG-containing composite significantly increased the micro-hardness of the adjacent demineralized dentin. ATR-FTIR revealed calcium phosphate peaks on the surface of the groups which used BAG-containing composite. FE-SEM revealed surface deposits partially occluding the dentin surface. No significant difference was found between SBF and PBS storage. Clinical Significance Bioactive glass is a potentially useful material for remineralization of tooth structure. When incorporated into a resin composite, it may aid in remineralizing the adjacent demineralized dentin, thus preventing further destruction of the tooth. Conclusion BAG-containing composites placed in close proximity can partially remineralize adjacent demineralized dentin.

This study compared the dentin bond strength of a new universal adhesive with that of contemporary multi-step dentin adhesives. Six experimental groups were prepared according to the adhesives used and their application modes: Optibond FL (OB), Adper Single Bond Plus (SB), One-Step Plus (OS), Clearfil SE Bond (CS), All-Bond Universal using etch-and-rinse mode (ABE), and AllBond Universal using self-etch mode (ABS). Micro-tensile bond strength (µTBS) and failure mode were evaluated for each group. The bonded interface was analyzed using transmission electron microscopy (TEM). As a result, µTBS of 6 experimental groups was followed as: OB=ABE=SE=ABS>SB>OS group. TEM micrographs of ABE and ABS groups revealed a homogenous adhesive layer formation. In conclusion, a new universal adhesive can make reliable bond to dentin, regardless of the application mode.

Heart failure and cardiac arrhythmias are the leading causes of mortality and morbidity worldwide. However, the mechanism of pathogenesis and myocardial malfunction in the diseased heart remains to be fully clarified. Recent compelling evidence demonstrates that changes in the myofilament Ca2+ sensitivity affect intracellular Ca2+ homeostasis and ion channel activities in cardiac myocytes, the essential mechanisms responsible for the cardiac action potential and contraction in healthy and diseased hearts. Indeed, activities of ion channels and transporters underlying cardiac action potentials (e.g., Na+, Ca2+ and K+ channels and the Na+-Ca2+ exchanger) and intracellular Ca2+ handling proteins (e.g., ryanodine receptors and Ca2+-ATPase in sarcoplasmic reticulum (SERCA2a) or phospholamban and its phosphorylation) are conventionally measured to evaluate the fundamental mechanisms of cardiac excitation-contraction (E-C) coupling. Both electrical activities in the membrane and intracellular Ca2+ changes are the trigger signals of E-C coupling, whereas myofilament is the functional unit of contraction and relaxation, and myofilament Ca2+ sensitivity is imperative in the implementation of myofibril performance. Nevertheless, few studies incorporate myofilament Ca2+ sensitivity into the functional analysis of the myocardium unless it is the focus of the study. Here, we describe a protocol that measures sarcomere shortening/re-lengthening and the intracellular Ca2+ level using Fura-2 AM (ratiometric detection) and evaluate the changes of myofilament Ca2+ sensitivity in cardiac myocytes from rat hearts. The main aim is to emphasize that myofilament Ca2+ sensitivity should be taken into consideration in E-C coupling for mechanistic analysis. Comprehensive investigation of ion channels, ion transporters, intracellular Ca2+ handling, and myofilament Ca2+ sensitivity that underlie myocyte contractility in healthy and diseased hearts will provide valuable information for designing more effective strategies of translational and therapeutic value.

PURPOSE We investigated the prognostic factors for distant metastasis (DM) in patients with locally advanced oropharyngeal cancer (OPC) treated with surgery and adjuvant radiotherapy with or without concurrent chemotherapy. MATERIALS AND METHODS Eighty-five patients treated between January 1995 and August 2014 were evaluated retrospectively. Data regarding the pathological tumour and nodal status, human papillomavirus (HPV) status, treatment characteristics, and pretreatment maximum standardized uptake value (SUVmax) of 18-fluoro-2-deoxyglucose positron emission tomography-computed tomography scan (18F-FDG PET-CT) were evaluated, and their influence on DM and survival outcomes were analyzed. RESULTS Median follow-up period was 48.0 months. Recurrence was observed in 20 patients, including locoregional recurrence and DM. DM was observed in 13 patients. A multivariate analysis confirmed that the presence of lymphovascular invasion (p=0.031), lower neck lymph node (LN) involvement (p=0.006), SUVmax ≥ 9.7 (p=0.014), and tumour size ≥ 3 cm (p=0.037) significantly affected DM. HPV status was not associated with DM. Perineural invasion (p=0.048), lower neck LNinvolvement (p=0.008), SUVmax ≥ 9.7 (p=0.019), and tumour size ≥ 3 cm (p=0.033) were also significant factors for the DM-free survival rate. CONCLUSION Lower neck LN involvement, high SUVmax in pretreatment 18F-FDG PET-CT, and large tumour size were predictive factors for DM in patients of OPC.