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Balancing selection is frequently invoked as a mechanism that maintains variation within and across populations. However, there are few examples of balancing selection operating on loci underpinning complex traits, which frequently display high levels of variation. We investigated mechanisms that may maintain variation in a focal polymorphism—leaf chemical profiles of a perennial wildflower (Boechera stricta, Brassicaceae)—explicitly interrogating multiple ecological and genetic processes including spatial variation in selection, antagonistic pleiotropy, and frequency-dependent selection. A suite of common garden and greenhouse experiments showed that the alleles underlying variation in chemical profile have contrasting fitness effects across environments, implicating two ecological drivers of selection on chemical profile: herbivory and drought. Phenotype-environment associations and molecular genetic analyses revealed additional evidence of past selection by these drivers. Together, these data are consistent with balancing selection on chemical profile, likely caused by pleiotropic effects of secondary chemical biosynthesis genes on herbivore defense and drought response., Editor’s Summary: Evidence for balancing selection acting on loci that control complex traits is limited. Here, the authors show evidence for past selection on chemical profile in a perennial wildflower by two ecological drivers, herbivory and drought, consistent with balancing selection on this trait.

The use of zirconia for implants and abutments has become more prevalent in implant dentistry as an alternative to the commonly used titanium implants, and peri-implant disease can still affect them. The erbium, chromium-doped:yttrium-scandium-gallium-garnet (Er, Cr:YSGG) laser has emerged as a promising treatment modality. The purposes of this in vitro study were to (1) determine the effects of the laser on the surface roughness of zirconia discs; (2) determine the extent of removal of a single species biofilm, E. coli, on the zirconia discs after applying the laser; (3) determine the amount of cell adhesion and proliferation utilizing fibroblasts on zirconia discs after treatment with the laser. All treatments will be compared with the commonly used ultrasonic instrumentation and hand scalers. For the first aim, gross examination revealed noticeable surface damage on the discs when using ultrasonic and scalers but not for the laser group. For surface roughness, the mean roughness was Pa= 0.623±0.185 μm, 0.762±0.421 μm, 0.740±0.214 μm, and 0.724±0.168 μm for control discs, and discs treated with either the Er,Cr:YSGG laser, ultrasonic instrumentation, and hand scalers respectively. There was no statistical significance among the groups (p=0.628). For bacteria decontamination, there was a statistical significance among the groups (p< 0.0001). Statistical significance was seen between the control group and each of the three treatment groups, favoring the treatment groups (p< 0.0001). Statistical significance was seen when comparing ultrasonic instrumentation and hand scalers (p= 0.000) as well as when comparing the Er,Cr:YSGG laser to hand scalers (p= 0.007), favoring both the ultrasonic instrumentation and Er,Cr:YSGG laser. No significance between the Er,Cr:YSGG laser group and the ultrasonic instrumentation group was noted (p =0.374). When comparing the cell attachment following treatment in each of the three groups and also without treatment (control), there was a statistical significance among the groups (p

There has been an increase in utilizing 3D printers in dental restorations. The purpose of the study is to compare mechanical properties of 3D-printed prostheses to those of self-cured and/or computer-aided design-computer-aided manufacturing (CAD-CAM) restorations. A metal master typodont was prepared for the mandibular left sextant with implant analogs embedded at the first premolar and first molar positions with a missing second premolar. Three-unit provisional fixed dental prosthesis (FDP) was designed utilizing the 3Shape tooth library and forty-five uniform specimens were fabricated with different materials: self-cured poly(methyl methacrylate) (PMMA) (N = 15), milled PMMA CAD-CAM blocks (N = 15) and 3D-printed resin (N = 15). All specimens were tested using an Instron machine at a crosshead speed of 0.5 mm/min by an axial load on the occlusal surface of the second premolar pontic site. Statistical analysis was completed with Shapiro-Wilk, ANOVA and Tukey post-hoc tests. Mean fracture force was 300.61 N, 294.64 N and 408.49 N for self-cured PMMA, milled PMMA and 3D-printed resin, respectively. Mean force at FDP fracture of 3D-printed resin was significantly greater than the mean fracture force of either self-cured (p = 0.016, 95% CI [17.86, 197.91]) or milled (p = 0.010, 95% CI [23.83, 203.88]) PMMA.

This investigation was designed to evaluate the reestablishment of bone-to-implant contact on infected dental implant surfaces following decontamination with an erbium, chromium:yttrium-scandium-gallium-garnet (Er,Cr:YSGG) laser and reconstructive therapy. Three patients presenting with at least one failing implant each were enrolled and consented to treatment with the Er,Cr:YSGG laser surface decontamination and reconstruction with a bone replacement allograft and a collagen membrane. The laser treatment was carried out at a setting of 1.5 W, air/water of 40%/50%, and pulse rate of 30 Hz. At 6 months, all three patients returned for the study. En bloc biopsy samples of four implants were obtained and analyzed. Two patients had excellent clinical outcomes, while one patient with two adjacent failing implants experienced an early implant exposure during the follow-up period. There was histologic evidence of new bone formation with two implant specimens and less bone gain with the others. Despite the small sample size, these were optimistic findings that suggested a positive role of Er,Cr:YSGG laser in debridement of a titanium implant surface to facilitate subsequent regenerative treatment. This investigation provides histologic evidence as well as encouraging clinical results that use of the Er,Cr:YSGG laser can be beneficial for treatment of peri-implantitis, but further long-term clinical studies are needed to investigate the treatment outcome obtained.

Acinetobacter baumannii is a prevalent pathogen that can rapidly acquire resistance to antibiotics. Indeed, multidrug-resistant A. baumannii is a major cause of hospital-acquired infections and has been recognised by the World Health Organization as one of the most threatening bacteria to our society. Resistance-nodulation-division (RND) type multidrug efflux pumps have been demonstrated to convey antibiotic resistance to a wide range of pathogens and are the primary resistance mechanism employed by A. baumannii. A component of an RND pump in A. baumannii, AdeT1, was previously demonstrated to enhance the antimicrobial resistance of Escherichia coli. Here, we report the results of experiments which demonstrate that wild-type AdeT1 does not confer antimicrobial resistance in E. coli, highlighting the importance of verifying protein production when determining minimum inhibitory concentrations (MICs) especially by broth dilution. Nevertheless, using an agar-based MIC assay, we found that propionylation of Lys280 on AdeT1 renders E. coli cells more resistant to erythromycin.

The objective of this study is to evaluate titanium decontamination after different protocols while assessing changes in surface roughness, chemical composition, and wettability.Ninety-six smooth (S) and 96 minimally rough (R) titanium microimplants were used. Pristine microimplants were reserved for negative control (S-nC/R-nC, n = 9), while the remaining microimplants were incubated in Escherichia coli culture. Non-decontaminated microimplants were used as positive control (S-pC/R-pC, n = 3). The other microimplants were divided into seven different decontamination protocols (12 S/R per group): 24% EDTA, 2% chlorhexidine (CHL), gauze soaked in 2% chlorhexidine (GCHL), gauze soaked in ultrapure water (GMQ), scaling (SC), titanium brush (TiB), and implantoplasty (IP). Contaminated areas were assessed by scanning electron microscope images, chemical composition by energy dispersive X-ray spectroscopy, wettability by meniscus technique, and roughness by an optical profiler.Higher residual bacteria were observed in R-pC compared with S-pC (P 0.0001). When comparing S and R with their respective pC groups, the best results were obtained with GCHL, SC, TiB, and IP, with no difference between these protocols (P 0.05). Changes in surface roughness were observed after all treatments, with S/R-IP presenting the smoother and a less hydrophilic surface (P 0.05). Apart from IP protocol, all the other groups presented a more hydrophilic surface in R than in S microimplants (P 0.003). All decontamination protocols resulted in a lower percentage of superficial Ti when compared with S/R-nC (P 0.002).All decontamination protocols resulted in changes in roughness, wettability, and chemical composition, but GCHL, SC, TiB, an IP presented the best decontamination outcomes.

This investigation was designed to evaluate the healing response of 9.3-μm CO2 laser-assisted periodontal therapy. Five patients presenting with moderate to severe periodontitis, with an initial pocket depth (PD) ≥ 5 mm and with teeth predetermined to be surgically extracted, were enrolled and consented to treatment with full-mouth CO2 laser-assisted therapy. The laser treatment was carried out in the Ultraguide Mode at a setting of 0.25-mm spot size, with an average power of 0.65 to 1.15 watts and 20% mist. The laser tip was passed from the gingival margin and down apically to the base of the pocket with a sweeping L motion. The teeth were intensely scaled with piezoultrasonic instrumentation afterwards. A second pass of the laser tip was performed for the study teeth. At 9 months, all patients were clinically reevaluated. For sites with an initial pocket depth of ≥ 7 mm, a mean PD reduction of 3.97 ± 1.36 mm and a mean clinical attachment level gain of 3.54 ± 1.54 mm were achieved, resulting in a mean PD of 3.91 ± 0.77 mm. En bloc biopsy samples of four teeth were obtained and analyzed; two demonstrated histologic evidence of new bone formation while the other two healed with a long junctional epithelium with minimal inflammatory infiltrate. Further long-term clinical studies are needed to investigate the treatment stability obtained with a 9.3-μm CO2 laser compared to conventional surgical therapy. Nevertheless, the encouraging clinical results indicated that adjunctive use of the 9.3-μm CO2 laser-assisted periodontal therapy can be beneficial for treatment of periodontally compromised patients.

Background/Aim: Casein kinase 1 epsilon (CK1ε) is a member of the casein kinase 1 family, which includes highly conserved and ubiquitous serine/threonine protein kinases. Recent research has revealed that CK1ε plays an important role in a variety of human cancer types; however, its role in human melanoma remains unclear. The aim of this study was to elucidate the clinical role of CK1ε in patients with melanoma. Patients and Methods: Samples from 34 patients with melanoma were analyzed by immunohistochemical staining. Formalin-fixed paraffin-embedded tissue microarrays were also examined by two histopathologists to assess CK1ε protein expression in humans. Results: Cytoplasmic CK1ε protein expression was significantly lower in tumor tissue than in normal tissue. Lack of cytoplasmic CK1ε protein was significantly correlated with distant metastasis (p=0.022) and poorer survival (p=0.030). However, Kaplan-Meier survival analysis revealed that elevated expression of cytoplasmic CK1ε protein was not significantly associated with the overall survival of patients with melanoma. Univariate and multivariate analyses demonstrated that lack of cytoplasmic CK1ε protein expression was related to distant metastasis (p

Background: Antinuclear antibody pattern recognition is vital for autoimmune disease diagnosis but labor-intensive for manual interpretation. To develop an automated pattern recognition system, we established machine learning models based on the International Consensus on Antinuclear Antibody Patterns (ICAP) at a competent level, mixed patterns recognition, and evaluated their consistency with human reading. Methods: 51,694 human epithelial cells (HEp-2) cell images with patterns assigned by experienced medical technologists collected in a medical center were used to train six machine learning algorithms and were compared by their performance. Next, we choose the best performing model to test the consistency with five experienced readers and two beginners. Results: The mean F1 score in each classification of the best performing model was 0.86 evaluated by Testing Data 1. For the inter-observer agreement test on Testing Data 2, the average agreement was 0.849 (κ) among five experienced readers, 0.844 between the best performing model and experienced readers, 0.528 between experienced readers and beginners. The results indicate that the proposed model outperformed beginners and achieved an excellent agreement with experienced readers. Conclusions: This study demonstrated that the developed model could reach an excellent agreement with experienced human readers using machine learning methods.

Irinotecan inhibits cell proliferation and thus is used for the primary treatment of colorectal cancer. Metabolism of irinotecan involves incorporation of β-glucuronic acid to facilitate excretion. During transit of the glucuronidated product through the gastrointestinal tract, an induced upregulation of gut microbial β-glucuronidase (GUS) activity may cause severe diarrhea and thus force many patients to stop treatment. We herein report the development of uronic isofagomine (UIFG) derivatives that act as general, potent inhibitors of bacterial GUSs, especially those of Escherichia coli and Clostridium perfringens. The best inhibitor, C6-nonyl UIFG, is 23,300-fold more selective for E. coli GUS than for human GUS (Ki = 0.0045 and 105 μM, respectively). Structural evidence indicated that the loss of coordinated water molecules, with the consequent increase in entropy, contributes to the high affinity and selectivity for bacterial GUSs. The inhibitors also effectively reduced irinotecan-induced diarrhea in mice without damaging intestinal epithelial cells., Hsien-Ya Lin, Chia-Yu Chen, Ting-Chien Lin and colleagues perform structure-guided modifications of the compound uronic isofagomaine in order to engineer a highly specific and potent inhibitor of gut bacterial β-glucuronidases (GUSs). The authors present eight crystal structures and demonstrate in vivo efficacy of the optimised C6-alkyl derivative inhibitor in mice models. This study may enhance the development of inhibitors of microbial GUS for use in colorectal cancer therapy to minimize the undesired side effects of irinotecan treatment.

Preterm labor is associated with inflammation and infection. The mechanisms underlying the role of omega-3 fatty acid in inflammasome activation and prevention of preterm labor remain unknown. We hypothesized that omega-3 fatty acid can reduce the rate of preterm birth induced by infection and trophoblast inflammation. In the present study, we found that inflammasome-related molecules and IL-1β in trophoblasts were activated by TNF-α derived from lipopolysaccharide (LPS)-stimulated THP-1 cell-conditioned medium (CM) and recombinant TNF-α protein. The results demonstrated that stimulation with TNF-α caused lysosomal rupture in trophoblasts, which accelerated cathepsin S (CTSS) diffusion from lysosomes to the cytosol and activated NLRP1 (nacht domain-leucine-rich repeat, and pyd-containing protein 1) and absent in melanoma 2 (AIM2) inflammasomes, thereby increasing IL-1β secretion. Moreover, in response to LPS challenge, TNF-α increased trophoblast cell death and decreased cell viability through inflammasome and CTSS activation. Stearidonic acid (SDA; 18:4n−3) and docosahexaenoic acid (DHA; 22:6n−3) inhibited inflammasome-related molecule synthesis and CTSS and caspase-1 activation, which further reduced the preterm delivery rate of pregnant mice induced by LPS (92.9 compared with 69.7% (DHA); 92.9 compared with 53.5% (SDA)). Higher expression of TNF-α, IL-1β, prostaglandin E2, and CTSS, but lower resolvin D1 expression, was observed in preterm pregnant mice than in controls. Similarly, resolvin D1 was highly expressed in women with term delivery compared with women with preterm delivery. Thus, SDA and DHA may attenuate macrophage-derived TNF-α inducing CTSS and inflammasome activation, IL-1β secretion, and placental trophoblast cell death. These functions are implicated in the preventive effects of SDA and DHA on preterm labor.

The goal of this multicenter randomized controlled study was to evaluate the effectiveness of a newly developed ionic-sonic electric toothbrush in terms of plaque removal and reduction of gingival inflammation. A total of 78 subjects from three dental centers were invited to join the study. They were randomized to receive either a manual toothbrush (control group) or an ionic-sonic electric brush (test group). Full-mouth prophylaxis and oral hygiene instructions based on the stationary bristle technique were provided 1 week prior to the baseline visit. At baseline and at each follow-up appointment, Plaque Index (PI) and Gingival Index (GI) were recorded. In addition, probing depth (PD) and bleeding on probing were recorded at baseline and at the last appointment (week 5). At completion of the study, subjects in the test group were given a questionnaire regarding their satisfaction with the toothbrush. Sixty-four subjects completed the study (control: 28; test: 36). The mean age of the subjects was 36.90 ± 12.19 years. No significant difference between the baseline and 5-week PD was found. Plaque removal efficacy and reduction in gingival inflammation were more significant for the test group at week 2. Both the control and test groups showed statistically significant improvement in PI and GI from baseline to week 5. The ionic-sonic toothbrush was more effective than manual toothbrush after a 1-week application.

In vitro evaluations are essential to gaining a better understanding of re-osseointegration, while reducing animal use and the overall costs of peri-implantitis studies. This pilot study evaluated preosteoblast migration from 3-D-printed scaffolds to decontaminated titanium microimplants, creating a system that tries to mimic the bone–implant interface. Smooth (S) and minimally rough (R) titanium microimplants were incubated in Escherichia coli cultures and divided into six groups according to the decontamination protocol applied: EDTA gel (EDTA); chlorhexidine (CHL); chlorhexidine-soaked gauze (GCHL); scaling (SC); titanium brush (TiB); and implantoplasty (IP). Pristine S and R microimplants were used as the controls (C). After the decontamination procedures, the microimplants were inserted in 3-D-printed polyurethane-based scaffolds previously inoculated with preosteoblast cell cultures. Cellular migration was assessed after 24, 72 and 120 hours by ATP quantification. At the 120-hour time point, there was no statistically significant difference between S-C, S-EDTA, S-CHL, S-GCHL and S-SC ( p > 0.05), and between R-C, R-EDTA and R-GCHL ( p > 0.05). The in vitro model developed in this pilot study successfully demonstrated cell migration on the different decontaminated surfaces. This methodology suggests that on smooth microimplants, EDTA, GCHL, SC and TiB decontamination may have a reduced impact on preosteoblast migration, while on minimally rough microimplants, EDTA and GCHL decontamination affected cell migration the least. However, when selecting a decontamination protocol, the effectiveness of the decontamination per se must also be considered.

The aim of this study was to evaluate the biological efficacy of a unique perpendicular protrusion of type-I collagen (Col-I) from TiO2 nanotubes (NT-EPF surface). We hypothesized that the NT-EPF surface would play bifunctional roles in stimulating platelet-mediated fibroblast recruitment and anchoring fibroblast-derived Col-I to form a perpendicular collagen assembly, mimicking the connective tissue attachment around natural teeth for the long-term maintenance of dental implants. Ti surface modification was accomplished in two steps. First, TiO2 nanotubes (NT) array was fabricated via anodization. Diameters and depths of NTs were controlled by applied voltage and duration. Subsequently, an electrophoretic fusion (EPF) method was applied to fuse Col-I into nanotube arrays in a perpendicular fashion. Surface wettability was assessed by contact angle measurement. The bioactivity of modified TiO2 surfaces was evaluated in terms of NIH3T3 fibroblast attachment, platelet activation, and collagen extension. Early attachment, aggregation, and activation of platelets as well as release of platelet-related growth factors were demonstrated on NT-EPF surfaces. Platelet-mediated NIH3T3 cells migration toward NT-EPF was significantly increased and the attached cells showed a typical fibrous morphology with elongated spindle shape. A direct linkage between pseudopod-like processes of fibroblasts to NT-EPF surfaces was observed. Furthermore, the engineered EPF collagen protrusion linked with cell-derived collagen in a perpendicular fashion. Within the limitation of this in vitro study, the TiO2 nanotube with perpendicular Col-I surface (NT-EPF) promoted better cell attachment, induced a strong platelet activation which suggested the ability to create a more robust soft tissue seal.

The goal of the present study was to evaluate human histologic healing of dental implants with a unique triangular neck design that is narrower than the implant body. Four patients in need of full-mouth reconstruction were recruited and received several implants to support a full-arch prosthesis. In each patient, two additional customized reduced-diameter implants were placed, designated to be harvested after 6 months of submerged healing. The eight harvested implants were all placed in healed edentulous maxillary or mandibular ridges. These implants were O 3.5 × 8 mm in size, and the final osteotomy drill allowed for the creation of a gap up to 0.2 mm in size between the coronal aspect of the triangular implant neck and the surrounding bone. At the end of the healing period, the implants were retrieved with the surrounding bone. Microcomputed tomography (μCT) was performed before processing the biopsy samples for undecalcified histologic exampination. Bone-to-implant contact (BIC) was measured from the μCT data and from buccolingual/buccopalatal and mesiodistal central histologic sections. All implant gaps were filled by mature remodeled bone. The mean BICs of the BL/BP and MD sections were 64.45% ± 6.86% and 65.39% ± 10.44%, respectively, with no statistically significant difference. The mean 360-degree 3D BIC measured all over the implant surface was 68.58% ± 3.76%. The difference between the BIC measured on the μCT and on the histologic sections was not statistically significant. The positive histologic results of the study confirmed the efficacy of this uniquely designed dental implant.

Slit proteins have been reported to act as axonal repellents in Drosophila; however, their role in the placental microenvironment has not been explored. In this study, we found that human placental multipotent mesenchymal stromal cells (hPMSCs) constitutively express Slit2. Therefore, we hypothesized that Slit2 expressed by hPMSCs could be involved in macrophage migration during placental inflammation through membrane cognate Roundabout (Robo) receptor signaling. In order to develop a preclinical in vitro mouse model of hPMSCs in treatment of perinatal infection, RAW 264.7 cells were used in this study. Slit2 interacted with Robo4 that was highly expressed in RAW 264.7 macrophages: their interaction increased the adhesive ability of RAW 264.7 cells and inhibited migration. Lipopolysaccharide (LPS)-induced CD11bCD18 expression could be inhibited by Slit2 and by hPMSC-conditioned medium (CM). LPS-induced activation of p38 and Rap1 was also attenuated by Slit2 and by hPMSC-CM. Noticeably, these inhibitory effects of hPMSC-CM decreased after depletion of Slit2 from the CM. Furthermore, we found that p38 siRNA inhibited LPS-induced Rap1 expression in RAW 264.7 cells, indicating that Rap1 functions downstream of p38 signaling. p38 siRNA increased cell adhesion and inhibited migration through reducing LPS-stimulated CD11bCD18 expression in RAW 264.7 cells. Thus, hPMSC-derived Slit2 may inhibit LPS-induced CD11bCD18 expression to decrease cell migration and increase adhesion through modulating the activity and motility of inflammatory macrophages in placenta. This may represent a novel mechanism for LPS-induced placental infection.

Gingival hyperpigmentation in the anterior region in people with a high smile line is an esthetic concern. Gingival depigmentation can be achieved through various procedures. Most techniques have shown successful short-term results; however, recurrence is observed in 50% of patients after 2 to 4 years. A 46-year-old woman presenting with a large black zone of gingival hyperpigmentation in the anterior maxilla was treated with scalpel blade excision. At 25 years after the depigmentation procedure, the patient displayed no recurrence.

The transcription factor glial cells missing 1 (GCM1) regulates trophoblast differentiation and function during placentation. Decreased GCM1 expression is associated with pre-eclampsia, suggesting that abnormal expression of GCM1 target genes may contribute to the pathogenesis of pregnancy complications. Here we identified a novel GCM1 target gene, synapse defective 1 (SYDE1), which encodes a RhoGAP that is highly expressed in human placenta, and demonstrated that SYDE1 promotes cytoskeletal remodelling and cell migration and invasion. Importantly, genetic ablation of murine Syde1 results in small fetuses and placentas with aberrant phenotypes in the placental–yolk sac barrier, maternal–trophoblast interface, and placental vascularization. Microarray analysis revealed altered expression of renin-1, angiotensin I converting enzyme 2, angiotensin II type 1a receptor, and membrane metalloendopeptidase of the renin–angiotensin system in Syde1-knockout placenta, which may compensate for the vascular defects to maintain normal blood pressure. As pregnancy proceeds, growth restriction of the Syde1−/− fetuses and placentas continues, with elevated expression of the Syde1 homologue Syde2 in placenta. Syde2 may compensate for the loss of Syde1 function because SYDE2, but not the GAP-dead SYDE2 mutant, reverses migration and invasion activities of SYDE1-knockdown JAR trophoblast cells. Clinically, we further detected decreased SYDE1 expression in preterm and term IUGR placentas compared with gestational age-matched controls. Our study suggests a novel mechanism for GCM1 and SYDE1 in regulation of trophoblast cell migration and invasion during placental development and that decreased SYDE1 expression is associated with IUGR. Copyright © 2016 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

BACKGROUND The aim of this systematic review and meta-analysis was to compare the clinical efficacy of the early dental implant placement protocol with immediate and delayed dental implant placement protocols. METHODS An electronic and manual search of literature was made to identify clinical studies comparing early implant placement with immediate or delayed placement. Data from the included studies were pooled and quantitative analyses were performed for the implant outcomes reported as the number of failed implants (primary outcome variable) and for changes in peri-implant marginal bone level, peri-implant probing depth, and peri-implant soft tissue level (secondary outcome variables). RESULTS Twelve studies met the inclusion criteria. Significant difference in risk of implant failure was found neither between the early and immediate placement protocols (risk difference = -0.018; 95% confidence interval [CI] = -0.06, 0.025; P = 0.416) nor between early and delayed placement protocols (risk difference = -0.008; 95% CI = -0.044, 0.028; P = 0.670). Pooled data of changes in peri-implant marginal bone level demonstrated significantly less marginal bone loss for implants placed using the early placement protocol compared with those placed in fresh extraction sockets (P = 0.001; weighted mean difference = -0.14 mm; 95% CI = -0.22, -0.05). No significant differences were found between the protocols for the other variables. CONCLUSIONS The available evidence supports the clinical efficacy of the early implant placement protocol. Present findings indicate that the early implant placement protocol results in implant outcomes similar to immediate and delayed placement protocols and a superior stability of peri-implant hard tissue compared with immediate implant placement.

Background In pancreatic cancer, toxicities associated with current chemotherapeutic regimens remain concerning. A modified combination of gemcitabine, S-1, and leucovorin (GSL) was used as the first-line treatment for newly diagnosed locally advanced or metastatic pancreatic adenocarcinoma patients. Methods GSL was administered every 2 weeks—intravenous gemcitabine 800 mg/m2 at a fixed-dose rate of 10 mg/m2/min on day 1 and oral S-1 (80–120 mg/day) plus leucovorin 30 mg twice daily on days 1–7. We retrospectively analyzed the feasibility of GSL and patient outcomes in three medical centers in Taiwan. Results Overall, 49 patients received GSL with a median follow-up of 24.9 months from May 2015 to March 2019. The median patient age was 68 years (range, 47–83 years), with a marginally higher number of females (57.1%). Among the 44 patients who underwent image evaluation, 13 demonstrated a partial response (29.5%) and 17 presented with stable disease (38.6%). The partial response rate and stable disease rate was 26.5% and 34.7%, respectively, in the intent-to-treat analysis. The median time-to-treatment failure was 5.79 months (95% C.I., 2.63–8.94), progression-free survival was 6.94 months (95% C.I., 5.55–8.33), and overall survival time was 11.53 months (95% C.I., 9.94–13.13). For GSL treatment, the most common grade 3 or worse toxicities were anemia (18.3%), neutropenia (6.1%), nausea (4.1%), and mucositis (4.1%). Treatment discontinuation was mostly due to disease progression (65.3%). Conclusions The modified GSL therapy can be a promising and affordable treatment for patients with advanced and metastatic pancreatic cancer in Taiwan. A prospective trial of modified GSL for elderly patients is currently ongoing in Taiwan.

The objective of this study was to investigate whether human placental multipotent mesenchymal stromal cell (hPMSC)-derived Slit2 and endothelial cell Roundabout (Robo) receptors are involved in placental angiogenesis. The hPMSC-conditioned medium and human umbilical vein endothelial cells were studied for Slit2 and Robo receptor expression by immunoassay and RT-PCR. The effect of the conditioned medium of hPMSCs with or without Slit2 depletion on endothelial cells was investigated by in vitro angiogenesis using growth factor-reduced Matrigel. hPMSCs express Slit2 and both Robo1 and Robo4 are present in human umbilical vein endothelial cells. Human umbilical vein endothelial cells do not express Robo2 and Robo3. The hPMSC-conditioned medium and Slit2 recombinant protein significantly inhibit the endothelial cell migration, but not by the hPMSC-conditioned medium with Slit2 depletion. The hPMSC-conditioned medium and Slit2 significantly enhance endothelial tube formation with increased cumulated tube length, polygonal network number and vessel branching point number compared to endothelial cells alone. The tube formation is inhibited by the depletion of Slit2 from the conditioned medium, or following the expression of Robo1, Robo4, and both receptor knockdown using small interfering RNA. Furthermore, co-immunoprecipitation reveals Slit2 binds to Robo1 and Robo4. Robo1 interacts and forms a heterodimeric complex with Robo4. These results suggest the implication of both Robo receptors with Slit2 signaling, which is involved in endothelial cell angiogenesis. Slit2 in the conditioned medium of hPMSCs has functional effect on endothelial cells and may play a role in placental angiogenesis.

Background: W e investigated the combined impact of abdominal obesity and low skeletal muscle mass on cardiovascular and total mortality in an elderly Asian population. Methods: A total of 1,485 elderly individuals (≥65 years) from Elderly Nutrition and Health Survey in Taiwan (1999–2000) were enrolled, and their survival status was followed using data from the National Death Registry. Skeletal muscle mass index (SMMI) was calculated by dividing skeletal muscle mass (kg) by height squared (m 2 ). Low skeletal muscle mass was defined as the first quartile of SMMI. Abdominal obesity (high triglycerides plus waist circumference [HTGWC]) was defined as triglycerides ≥150 mg/dL and w aist circumference ≥90 cm (men) and ≥80 cm (women). The Cox proportional hazard model was used to evaluate the combined impact of abdominal obesity and low SMMI on cardiovascular and total mortality. Results: During follow -up (median 9.2 years), one third (n = 493) of subjects died from any cause, of which 34% (n = 168) were cardiovascular-related. Total and cardiovascular mortality were 4.2 and 1.4 per 100 person-years, respectively. Low SMMI and HTGWC were independently associated with total mortality in men, but only low SMMI was significantly associated in women. Those with both HTGWC and low SMMI had the highest mortality risk, with the cardiovascular mortality risk increased by >6.8-fold and 3.2-fold in men and women, respectively, compared with controls having normal SMMI and TGWC. Conclusions: Elderly indi viduals with abdominal obesity and low skeletal muscle mass have higher all-cause and cardiovascular mortality risk.

Background Chylous leakage is a well-recognized but rare complication of head and neck surgery, affecting approximately 1% to 2.5% of head and neck dissections. It is a potentially life-threatening condition characterized by electrolyte imbalance, immunosuppression, delayed wound healing, risk of infection, and generalized sepsis. Management can be problematic and prolonged. Methods We present a case of refractory cervical chylous leakage after neck dissection treated with ultrasound-guided intranodal lymphangiography. Results Ultrasound-guided intranodal lymphangiography alone resulted in rapid and complete resolution of chylous leakage with minimal morbidity. Conclusion Based on our clinical experience and after a thorough literature review, we propose that ultrasound-guided intranodal lymphangiography with contrast agent could be considered a viable therapeutic option for persistent chylous leakages in selected patients. © 2015 Wiley Periodicals, Inc. Head Neck 38: E54–E60, 2016

Epithelial attachment via the basal lamina on the tooth surface provides an important structural defence mechanism against bacterial invasion in combating periodontal disease. However, when considering dental implants, strong epithelial attachment does not exist throughout the titanium-soft tissue interface, making soft tissues more susceptible to peri-implant disease. This study introduced a novel synthetic peptide (A10) to enhance epithelial attachment. A10 was identified from a bacterial peptide display library and synthesized. A10 and protease-activated receptor 4-activating peptide (PAR4-AP, positive control) were immobilized on commercially pure titanium. The peptide-treated titanium showed high epithelial cell migration ability during incubation in platelet-rich plasma. We confirmed the development of dense and expanded BL (stained by Ln5) with pericellular junctions (stained by ZO1) on the peptide-treated titanium surface. In an adhesion assay of epithelial cells on A10-treated titanium, PAR4-AP-treated titanium, bovine root and non-treated titanium, A10-treated titanium and PAR4-AP-treated titanium showed significantly stronger adhesion than non-treated titanium. PAR4-AP-treated titanium showed significantly higher inflammatory cytokine release than non-treated titanium. There was no significant difference in inflammatory cytokine release between A10-treated and non-treated titanium. These results indicated that A10 could induce the adhesion and migration of epithelial cells with low inflammatory cytokine release. This novel peptide has a potentially useful application that could improve clinical outcomes with titanium implants and abutments by reducing or preventing peri-implant disease.

The volumes of alkaline (pH > 10), Bacillus thuringiensis Cry protein-solubilizing buffers imbibed by fall armyworm larvae in droplet feeding assays were determined. The buffers differed in the presence or concentration of key ingredients, including buffering agents, chelating agents, reducing agents, and protease inhibitors. For both first and second instar larvae, the buffer used had a significant effect on the volume imbibed. The study showed that the droplet feeding method is compatible with Cry protein-solubilizing buffers, but that it is important to determine the volume imbibed for every buffer used in dose-dependent bioassays in order to reduce dose errors.

Background The effect of different surgical techniques for ridge preservation on soft tissue parameters has seldom been investigated. The objective of this study was to compare the effect of two different ridge preservation techniques on soft and hard tissue dimensions. Methods Thirty patients requiring tooth extraction were randomly allocated to either control group C (allograft covered with a non-crosslinked collagen membrane with primary closure) or experimental group E (allograft covered with cross-linked collagen membrane left exposed). Sites were surgically re-entered at 6 months. Soft and hard tissue measurements, cone beam computed tomography (CBCT), and cast measurements were taken at baseline and 6 months. Results Twenty-eight patients were included in this analysis. When the two treatment groups were compared, the width of the buccal keratinized tissue in the E group showed an increase of 0.43 ± 0.42 mm compared to net loss of 1.57 ± 0.51 mm for the C (P = 0.006). Similarly, buccal tissue thickness has increased in the E group 0.46 ± 0.22 mm compared to a loss of 0.15 ± 0.23 mm in the C group (P = 0.068). Volumetric assessment of the changes in the alveolar ridge for the E group showed a slight decrease (68.3 ± 17 mm3) whereas the C group has experienced almost double this loss (107.5 ± 11 mm3; P = 0.07). Crestal width, measured on the CBCT scan, has shown significant reduction in the C group (4.18 ± 0.56 mm) compared to only 1.74 ± 0.4 mm in the E group (P = 0.003). Conclusion Crosslinked collagen membrane with allograft placed intentionally non-submerged resulted in better preservation of the keratinized tissues (width and thickness) with similar and at times better osseous preservation following extraction.

STEAP1 (six transmembrane epithelial antigen of the prostate 1) is a transmembrane protein that functions as a potential channel or transporter protein. It is overexpressed in certain cancers and is viewed as a promising therapeutic target. However, the prognostic role of STEAP1 is still controversial, and no role for STEAP1 has yet been indicated in colorectal cancer. The aim of this study was to investigate the possible association of STEAP1 expression with colorectal cancer prognosis. STEAP1 expression was analyzed by immunohistochemical staining of a tissue array of 165 cancer specimens from primary colorectal cancer patients. The mean and medium follow-up times after surgery were 5.1 and 3.9 years, respectively. A total of 139 patients died during the 13 years of follow-up in the survey period. The prognostic value of STEAP1 with respect to overall survival was analyzed by Kaplan-Meier analysis and Cox proportional hazard models. In total, 164 samples displayed detectable STEAP1 expression in the cytoplasm and membrane. Low STEAP1 expression was correlated with poor overall survival (five-year survival: 33.7% vs. 57.0%, low expression vs. high expression, p = 0.020). Accordingly, multivariate analysis identified low STEAP1 expression as an independent risk factor (hazard ratio = 1.500, p = 0.018), especially in elderly patients or those with late stage cancers, late T values, and early N values. We suggest that analysis of STEAP1 expression by immunohistochemical staining could serve as an independent prognostic marker for colorectal patients. This finding should be validated by other investigative groups.

Over the past two decades, studies have widely examined the effectiveness of pay-for-performance (P4P) programs by conducting biochemical tests and assessing complications; however, the reported effectiveness of such programs among participants selected through purposeful sampling is controversial. Therefore, the objective of the current study was to analyze the effectiveness of a P4P program on patients' prognoses, including hospitalization for chronic diabetic complications, and all-cause mortality during specific follow-up years by using a nationwide population-based database in Taiwan. Based on 125,315 newly diagnosed type 2 diabetes patient cohort during 2002-2006, two control sets were designed by propensity-score-matching strategy according to participation of P4P program and followed up to 2012. The results indicated that full participants demonstrated the lowest risks of developing complications and all-cause mortality compared with nonparticipants. These findings confirm the long-term effect of P4P programs on full participants and reveal that this effect is not due to confounding variables. The results indicate the importance of performance management and adherence to interventions for patients with chronic diseases in a long-term observation. Comprehensive and continuous care is suggested to improve patient prognosis and quality of care.

Background: Impaired cognition increases mortality in the aged. It is unclear how dietary quality might affect this relationship. Objective: To examine how dietary diversity and cognition might interact to determine survival. Design: In a Nutrition and Health Survey in Taiwan (NAHSIT 1999-2000), 1,839 representative elderly were followed for mortality up to 10 years. The dietary quality measure was a dietary diversity score (DDS, range: 0-6) to present six food groups (dairy, meat, rice and grains, fruit, vegetable,fat and oil) derived from a 24-h dietary recall. Cognitive function was evaluated by the validated Short Portable Mental Status Questionnaire (SPMSQ). Results: Those with cognitive impairment (SPMSQ≥3 errors) had 2.56 (95% confidence intervals (CI), 1.99- 3.28) times the all-cause-mortality risk of those with intact cognition. After control for potential confounders, the adjusted hazard ratio (HR) remained significant (1.46, 95% CI: 1.06-2.02). Significant interactions for DDS and cognition were found (p

Introduction Population aging is set to increase the elder dependency ratio, causing a rapid increase in healthcare expenditures and financial burden on the government. This study aims to construct an aging score from age-related diseases and to perform longitudinal analyses to examine the association between personal income and aging. Methods A total of 86,838 subjects drawn from a community with 384,617 residents in northern Taiwan were examined. Personal income and aging-related diseases were measured from the National Health Insurance database during 1997–2010. Multiple linear regression and stratified analyses were used. Results Our results indicate that subjects with low personal income (defined as monthly income ≤610 USD) can have high aging scores as calculated by selected aging-related diseases. In stratified analyses, moreover, subjects with low personal income tended to have higher aging scores across different anthropometric groups. The association was particularly evident in subjects >60 years old, and they were noted across multiple strata, including those of catastrophic illness as well as type of primary care setting. Conclusions This study found evidence for association between low income and aging in a population with compulsory health insurance program, where accessibility barriers were low in terms of health care resources and personal socioeconomic status. The economic inequity causes difference in aging process, which deserves for future interventions.

Rac GTPase activating protein 1 (RACGAP1) plays a regulatory role in initiation of cytokinesis, control of cell growth and differentiation, and tumor malignancy, making it a potential prognostic biomarker. RACGAP1 is present in the nucleus, but a diffuse distribution in the cytoplasm also occurs. The aim of this study was to determine the impact of nuclear and cytoplasmic expression of RACGAP1 on clinical outcome to provide further evidence of a role in colorectal cancer. RACGAP1 expression was analyzed by immunohistochemistry in 166 cancer specimens from primary colorectal cancer patients. The mean follow-up time after surgery was 5.4 years (range, 0.01-13.10 years). The prognostic value of RACGAP1 on overall survival was validated by Kaplan-Meier analysis and Cox regression models. RACGAP1 is expressed in colorectal specimen and is present in both the nucleus and cytoplasm in different amounts. Colorectal cancer patients had opposite prognoses depending on the site of RACGAP1 expression. Patients with high nuclear RACGAP1 expression had poor outcomes, whereas those with high cytoplasmic RACGAP1 expression had favorable prognosis (P = .003 and P = .001, respectively). Patients with low nuclear but high cytoplasmic RACGAP1 expression had better survival compared with those with other combinations (P < .001). We suggest that RACGAP1 expression levels in the nucleus and cytoplasm, determined by immunohistochemical staining, predict opposite clinical outcomes and that both could be independent prognostic markers for colorectal cancer.

Background Many studies have investigated the association between markers for peripheral inflammation and risk of dementia, but the results have been conflicting. We aimed to evaluate the association between a specific inflammation marker, C-reactive protein (CRP), and dementia in an elderly Asian community cohort. Methods The cohort included 1436 individuals (ages 65 and older) from a national representative sample in Taiwan. Dementia incidence was identified using International Classification of Diseases , Ninth Revision codes for vascular dementia, Alzheimer disease, and nonvascular dementia. Baseline characteristics and CRP levels were determined. A Cox proportional hazard model and Fine and Grays model were adjusted for stroke and competing risk of death to estimate the association between inflammation and development of dementia. Results During 11.04 years (median) of follow-up, 607 individuals (50.77%) died and 260 individuals (18.11%) were diagnosed with dementia. Those with high CRP had a 55% higher risk of dementia (hazard ratio 1.55; 95% confidence interval 1.21–2.00) compared with those with normal CRP. After adjusting for possible confounding cardiovascular risk factors, high CRP was independently associated with vascular dementia but not Alzheimer disease. Conclusions In this prospective study of an elderly Asian community cohort with more than 10 years of follow-up, the baseline serum CRP level was associated with future development of vascular dementia, but not Alzheimer disease after adjusting for common cardiovascular risk factors, stroke, and competing risk of death.

The X-linked DEAD-box RNA helicase DDX3 (DDX3X) is a multifunctional protein that has been implicated in gene regulation, cell cycle control, apoptosis, and tumorigenesis. However, the precise physiological function of Ddx3x during development remains unknown. Here, we show that loss of Ddx3x results in an early post-implantation lethality in male mice. The size of the epiblast marked by Oct3/4 is dramatically reduced in embryonic day 6.5 (E6.5) Ddx3x-/Y embryos. Preferential paternal X chromosome inactivation (XCI) in extraembryonic tissues of Ddx3x heterozygous (Ddx3x-/+) female mice with a maternally inherited null allele leads to placental abnormalities and embryonic lethality during development. In the embryonic tissues, Ddx3x exhibits developmental- and tissue-specific differences in escape from XCI. Targeted Ddx3x ablation in the epiblast leads to widespread apoptosis and abnormal growth, which causes embryonic lethality in the Sox2-cre/+;Ddx3xflox/Y mutant around E11.5. The observation of significant increases in γH2AX and p-p53Ser15 indicates DNA damage, which suggests that loss of Ddx3x leads to higher levels of genome damage. Significant upregulation of p21WAF1/Cip1 and p15Ink4b results in cell cycle arrest and apoptosis in Ddx3x-deficient cells. These results have uncovered that mouse Ddx3x is essential for both embryo and extraembryonic development.