Survival and success rates of soft-milled cobalt-chromium-ceramic full-arch screw-retained implant-supported prostheses: a 2- to 7-year follow-up retrospective study.

OBJECTIVES: To retrospectively assess implant and prosthesis survival and success and patient satisfaction in edentulous patients treated with soft-milled cobalt-chromium-ceramic full-arch screw-retained implant-supported prostheses (SCCSIPs) over a mean clinical follow-up period of 4 years. MATERIALS AND METHODS: Twenty-one edentulous patients, aged 47-80 years, underwent restoration with 27 SCCSIPs (210 implants, 349 crown units). After definitive prosthesis insertion, patients participated in a yearly dental check-up recall program, including clinical and radiographic examinations. All supporting implants were examined for biological and technical complications, and the restorations were evaluated using modified California Dental Association (CDA) criteria. Life table and Kaplan-Meier survival analyses were performed. Patient satisfaction regarding chewing, esthetics, comfort, and phonetics was evaluated using a visual analog scale (VAS). RESULTS: After a mean of 4 years, two implants (3.8%) failed, with no effect on prosthesis survival. The cumulative survival rate was 98.6% and 100% for implants and prostheses, respectively. The most frequent minor biological complication was soft tissue recession (11%). Porcelain fracture was the only major technical complication (0.6% of crown units), while the only minor technical complication was porcelain chipping (4% of crown units), which required only polishing. Overall, 66.7% of the prostheses were free of technical complications. Patient satisfaction was high to very high. Less satisfaction was noted with comfort due to soft tissue recession and food impaction. CONCLUSIONS: Within the limitations of this study, we conclude that the survival and success rates of SCCSIPs in edentulous patients were excellent. CLINICAL RELEVANCE: SCCSIPs should be considered when planning metal-ceramic implant-supported restorations.

The effect of gingival wall location on the marginal seal of class ii restorations prepared with a flowable bulk-fill resin-based composite.

SureFil SDR is a flowable resin-based composite that allows a single incremental bulk placement. The marginal seal of SureFil SDR at the gingival margins of class II restorations located apical to the cemento-enamel-junction (CEJ) has not been adequately evaluated compared to those located occlusal to the CEJ. Forty class II cavities were prepared in human molars. The gingival margins of 20 preparations were located 0.5 mm occlusal to the CEJ, and the other 20 preparations were located 0.5 mm apical to the CEJ. The cavities surfaces were bonded with XenoV dental adhesive and filled with SDR in one bulk increment up to 4 mm, after which they were covered with CeramX. The teeth were subjected to thermo-and load-cycling, and their gingival margins were exposed to 0.5% basic-fuchsin solution. The specimens were sectioned mesio-distally and scored for microleakage. A Wilcoxon test for pairwise comparison was performed to determine significance. Dye penetration was observed in 30% of the 20 restorations with cavo-surface margins located occlusal to the CEJ and in 55% of the 20 restorations with cavo-surface margins located apical to the CEJ. The bulk-fill flowable resin base SureFil SDR with XenoV dental adhesive provided a better marginal seal in class II restorations with gingival margins above the CEJ compared to restorations with gingival margins below the CEJ. SDR should not be recommended for class II cavity preparations with gingival margins located below the CEJ.

Survey results of currently used techniques and materials by dentists for fabrication of complete removable dental prostheses.

The present study was based on the screening of procedures and materials used by dentists in constructing complete removable dental prostheses, considering their professional background and years of experience. Questions arose regarding differences and similarities within the local dental society, of which approximately 75% were trained in Europeans countries with differences in academic background, and compared to international dentistry. A questionnaire web site was used in order to anonymously collect answers to the survey by general practicing and specialist dentists. The survey contained 28 multiple choice questions that were based on 5 parameters: years of experience, location of dental studies, specialization, location and field of specialization. One hundred one survey sheets were collected. The results show statistical significance (p less than 0.05) for the following comparisons: practicing time had an impact on practitioner-technician interaction, while the level of qualification had an effect on articulator and face-bow usage. Location of dental studies, specialization and residency had impact on final impression materials, teeth set-up as well as tooth shape and color selection. The differences found between dentists’ working techniques set the ground for future research that should focus on rating the success of the different methods towards a goal of creating recommendations for an ideal set of working techniques.

The effect of enriching denture adhesives with chlorhexidine diacetate on the proliferation of Candida albicans: an in vitro analysis.

Denture adhesives are widely used in dentistry. The growth of Candida albicans on the denture base is common on both jaws, particularly in elderly patients. A strong correlation has been found between the growth of Candida albicans and dentures, potentially leading to denture stomatitis. Recent studies show that the addition of chlorhexidine diacetate (CHDA) to denture adhesives can have a protective effect against denture stomatitis. The aim of this research is to examine the antifungal properties of three denture adhesives when CHDA is added. We hypothesize that the addition of CHDA to denture adhesives will result in antifungal activity against Candida albicans. Three denture adhesive materials (Corega, Protefix and Kukident) were prepared with and without the addition of CHDA in order to measure whether CHDA increased the resistance of the adhesives to the growth of Candida albicans. Dynamic and static intercellular changes were assessed through direct contact test (DCT) and confocal microscopy. The tested materials were evaluated immediately after setting and at 14 days post-setting. When combined with CHDA, all three denture adhesive materials decreased Candida albicans growth compared to the control (p less than 0.0001). This inhibition of growth was more prominent in the first 24 h. The confocal microscopy results supported the DCT findings. The addition of CHDA to denture adhesive pastes inhibits growth of Candida albicans, and may potentially reduce the incidence of denture stomatitis. Further studies are warranted.

Temperature changes of one-piece implants during the setting of acrylic resin temporary crown. The effect of implant diameter. An in vitro study.

The purpose of this work is to evaluate changes in temperature of one-piece titanium implant surface during the setting of acrylic resin temporary crowns and to correlate thermal changes to implant diameter. Thirty-three one-piece implants (ARRP, Alpha-Biotec) were divided into 3 groups according to diameter size (G1=3 mm, G2=3.3 mm, G3=3.6 mm). Implants were mounted on an acrylic glass apparatus. Thermocouples were positioned at the most coronal thread. Lower incisor temporary polycarbonate crowns were filled with 80 µL of self-curing acrylic resin and positioned immediately on the implant abutment. Thermal changes of the implant surface were recorded continuously for 10 min. Data were statistically analyzed using one-way analysis of variance. The mean initial temperature (C0) of groups G1, G2 and G3 was similar (24.79±0.78ºC, 25.26±0.63ºC, 24.97±1.06ºC, respectively). The setting of the acrylic resin temporary crown resulted in a significant increase in the implant surface temperature of all groups. The mean thermal amplitude (ΔC) for groups G1, G2 and G3 were 6.79±1.02ºC, 6.61±0.94ºC, 6.65±1.26ºC, respectively. The mean time to maximum temperature (Tmax) for groups G1, G2 and G3 were 337.38±42.91 sec, 324.69±41.46 sec and 317.98±37.91 sec respectively (P>0.05). Direct application of auto-polymerizing resin to the titanium abutment of one-piece implants significantly increased the cervical implant surface temperature. Implant diameter did not influence the temperature changes.

Efficacy of XP-endo finisher files in the removal of calcium hydroxide paste from artificial standardized grooves in the apical third of oval root canals.

AIM: To compare the efficacy of the XP-endo finisher file (XP) (FKG Dentaire, La Chaux de Fonds, Switzerland) to that of passive ultrasonic irrigation (PUI) and conventional syringe and needle irrigation (SNI) in the removal of calcium hydroxide paste from an artificial standardized groove in the apical third of root canals. METHODOLOGY: The root canals of 68 mandibular incisors with single oval canals were prepared using Mtwo instruments (VDW GmbH, Munich, Germany) up to size 40, .04 taper. Each tooth was split longitudinally, and in one half of the root, a standardized groove was prepared in the apical part of the specimen. The grooves were filled with Ca(OH)2 , and the root halves were reassembled. The roots were randomly divided into two control groups (n = 4) and three experimental groups (n = 20) according to the Ca(OH)2 methods used: XP, PUI and SNI. The amount of remaining medicament was evaluated under X25 magnification using a 4-grade scoring system. Kappa values were calculated for intra- and interobserver agreement evaluation. The differences in the Ca(OH)2 scores amongst the different groups were analysed using the Kruskal-Wallis test. RESULTS: None of the tested methods could completely clean the Ca(OH)2 from the artificial standardized groove in the apical third of the root canals. XP and PUI removed significantly more Ca(OH)2 than SNI (P < 0.001), with no significant differences between them (P = 0.238). CONCLUSIONS: XP and PUI were more effective in removing Ca(OH)2 from artificial standardized grooves in the apical third of root canals than SNI.

First trimester human placental factors induce breast cancer cell autophagy.

Placental factors, progesterone included, facilitate breast cancer cell line (BCCL) motility and thus may contribute to the advanced breast cancer found during pregnancy. Cancer and placental implantations are similar; the last is accompanied by extravillous trophoblast cell invasion and autophagy which are interlinked. We aimed to analyze the effect of first trimester human placenta on BCCL autophagy. BCCLs (MCF-7/T47D) were cultured with placental explants (60 h) or placental supernatants (24 h). Following cultures, BCCLs were sorted out for RNA/protein extraction. RNA served for microarray/qPCR (BNIP3) and protein for Western blot (HIF1α, LC3BII) analyses. Inhibitors were added to the placenta-MCF-7 coculture or placental supernatants (autophagy inhibitor-3MA, progesterone receptor (PR) inhibitor-RU486, and HIF1α inhibitor-Vitexin) in order to evaluate their effects on BCCL motility and LC3BII/HIF1α expression. LC3BII (an autophagy marker) expression was elevated in BCCLs following placental explant coculture and exposure to placental supernatants. The autophagy inhibitor (3MA) repressed the placenta-induced MCF-7/T47D migration, establishing a connection between BCCL autophagy and migration. Microarray analysis of MCF-7 following placenta-MCF-7 coculture showed that "HIF1α pathway," a known autophagy facilitator, was significantly manipulated. Indeed, placental factors elevated HIF1α and its target BNIP3 in the BCCLs, verifying array results. Lastly, PR inhibitor reduced HIF1α expression and both PR and HIF1α inhibitors reduced MCF-7 LC3BII expression and motility, suggesting involvement of the PR-HIF1α axis in the autophagy process. Placental factors induced BCCL autophagy that is interlinked to their motility. This suggests that autophagy-related molecules may serve as targets for therapy in pregnancy-associated breast cancer.

The effect of heat shock protein 27 on extravillous trophoblast differentiation and on eukaryotic translation initiation factor 4E expression.

Heat shock protein (HSP27) is expressed in human placentae. Previously, we showed that HSP27 is expressed in the villous cell column of first trimester placental explants and in extravillous trophoblast (EVT) cells. EVT differentiation is accompanied by increased motility, matrix metalloproteinase (MMP) activity, decreased proliferation and expression of specific markers such as HLAG and CD9. HSP27 regulates cell apoptosis, migration, protein stability and the availability of eukaryotic translation initiation factors, such as eukaryotic translation initiation factor 4E (eIF4E). eIF4E supports trophoblast cell proliferation and survival. We wanted to explore the effect of HSP27 silencing on trophoblast cell phenotype, EVT markers and eIF4E expression and regulators [4E-binding protein (4E-BP1) and MAP kinase-interacting kinase (MNK1)]. This study evaluated the effect of HSP27 siRNA on placental explant and HTR-8/SVneo migration, MMP activity/mRNA, cell death, cell cycle, HLAG/CD9 levels, and eIF4E and its regulators' total and phosphorylated levels. Furthermore, we evaluated HSP27 levels in placentae exposed to ribavirin, which triggers EVT differentiation. We found that HSP27 silencing increased cell death in HTR-8/SVneo and placental explants. Furthermore, it reduced HTR-8/SVneo migration and EVT outgrowth from the explants (P < 0.05), MMP2 activity and expression of EVT markers HLAG and CD9 (in placental explants and HTR-8/SVneo, respectively, P < 0.05). Induction of EVT differentiation by ribavirin elevated HSP27 levels. Finally, HSP27 silencing in both HTR-8/SVneo and placental explants reduced eIF4E levels (33 and 28%, respectively, P < 0.05) and the levels of its regulators 4E-BP1 and MNK1 (37 and 32%, respectively, done on HTR-8/SVneo only), but not their phosphorylated forms. Altogether, our results suggest that HSP27 contributes to EVT cell differentiation.

The effects of in vitro fluoride mouth rinse on the antibacterial properties of orthodontic cements.

OBJECTIVES: To investigate the ability of orthodontic cements to regain their antibacterial effect after aging for 1 month, followed by 2 weeks of fluoride 'recharging' through daily fluoride rinse. MATERIALS AND METHODS: Four orthodontic cements were tested: composite resin-based materials (Transbond XT and Transbond Plus), a conventional glass ionomer cement (CX Plus) and dual-cured resin-reinforced glass ionomer cement (Fuji ORTHO LC) by direct contact test. After polymerization and a 30-day aging process, the samples were rinsed daily with 0.05% NaF solution for 14 days. Twenty-four hours after the last fluoride rinse, Streptococcus mutans cells (approximately 1 x 10(6) ) were placed on the surface of each sample for 1 h at 37°C to establish direct contact. Bacterial growth was monitored for 24 h by temperature-controlled spectrophotometry. Similar experiments were conducted after aging for 48 h and 72 h after the last fluoride rinse. One-way anova, two-way anova, and Tukey's multiple comparison test were applied to the data. RESULTS: Twenty-four hours after the last fluoride rinse, the resin-modified glass ionomer and the glass ionomer showed potent antibacterial properties, whereas the composite-based material Transbond XT encouraged bacterial growth. After 72 h, only Fuji Ortho LC maintained its bacteriostatic properties while all the other tested materials showed no antibacterial activity. CONCLUSIONS: Fourteen days daily fluoride rinse temporarily revives the antibacterial properties of the resin-modified glass ionomer cement and glass-ionomer-based cement.

Hormone-dependent placental manipulation of breast cancer cell migration.

BACKGROUND: Breast cancer during pregnancy is often more advanced than in non-pregnant women. Nevertheless, no case of metastasis inside the placenta has been reported. Previously, we showed that placental-explants eliminated breast cancer cells from their surroundings, due to cell-death and elevated migration. Our objective was to find the underlying mechanisms of these phenomena. METHODS AND RESULTS: Our model contained Michigan Cancer Foundation 7 (MCF7) or T47D cells co-cultured with and without human placental explants. Microarray analysis, validated by quantitative PCR, of MCF7 following their placental co-culture suggested activation of estrogen (E(2)) signaling. As extensive cross-talk exists between E(2) and progesterone, their involvement in mediating placental effects on breast cancer cells was tested. Indeed, addition of E(2) and progesterone receptor (ER and PR) inhibitors to the co-culture system reduced cancer cell motility, yet did not alter cell-cycle or death. E(2) and progesterone concentrations in placental media were found to be similar to those of early pregnancy blood levels. Interestingly, placental-breast cancer co-culture media contained lower progesterone (P < 0.05) and higher E(2) (200%, P < 0.05) levels than placentae cultured separately. Placental supernatant and E(2) and progesterone at placental levels were sufficient to increase MCF7 and T47D migration and invasion (P < 0.05), yet did not alter MCF7 cell-cycle or death. Furthermore, placental supernatant elevated p38 and Jun N-terminal kinase (JNK) phosphorylation in both cell lines (P < 0.05). Inhibitors of JNK, ER and PR reversed MCF7 and T47D motility induced by the placenta, suggesting their involvement. CONCLUSIONS: We suggest that E(2) and progesterone contribute to cell migration away from placental areas. We hypothesize that they may increase metastatic spread to other organs in pregnancy.

The efficacy of five techniques for removing root filling material: microscopic versus radiographic evaluation.

AIM: To test and compare the efficacy of five methods for the removal of root filling material and to test the hypothesis that radiographs fail to represent the real extent of remaining material on canal walls. METHODOLOGY: Fifty maxillary anterior single-rooted teeth with straight root canals were selected. The coronal third of each root canal was prepared with Gates-Glidden drills to number 3, whilst the apical two-thirds were prepared with manual K-files to size 40. Root fillings were performed using lateral compaction with gutta-percha and AH-26. After full setting, the coronal third of the root filling was removed with Gates-Glidden drills and the teeth divided into five groups (n=10). The remaining root filling material was then removed with either Hedström files and chloroform (25 µL), using size 40 as the last file, SafeSider files, using a NiTi Pleezer reamer with a 0.06 taper followed by size 40 reciprocating file, with or without chloroform, or ProTaper Universal retreatment files (D2, D3) with or without chloroform. Reaching working length with no more gutta-percha on the last file was defined as the endpoint for all procedures. The presence of remaining filling material was first evaluated radiographically and then by the microscopic evaluation of split roots. The time required to accomplish the procedure was also recorded. anova and anova with repeated measures were used for statistical analysis of the results. RESULTS: Overall, 11-26% of the canal wall remained covered with filling material; no significant difference was found between the groups. The mechanized methods were faster than manual removal of filling material (P < 0.01); the use of solvent did not speed up the mechanized procedures. Radiographic evaluation failed to adequately and reliably detect the extent of filling material remaining on the canal walls, which was later observed by microscopic evaluation. CONCLUSIONS: All methods left root canal filling material on the canal walls. Radiographic evaluation failed to detect the extent of remaining root filling material, which could only be detected using microscopy.

Breast cancer characteristics are modified by first trimester human placenta: in vitro co-culture study.

BACKGROUND: Pregnant women with breast cancer present with a more advanced disease compared with non-pregnant women. Nevertheless, breast cancer metastasis to the placenta is rare. Trophoblast/tumor implantations share the same biochemical mediators, while only the first is stringently controlled. We hypothesized that the same mechanisms that affect/restrain placental implantation may inhibit metastatic growth in the placenta. We aimed to analyze the effects of human placenta on breast cancer cells. METHODS: First trimester human placental explants were co-cultured with MCF-7/T47D-eGFP tagged cells. Following culture, placenta/cancer cells/both were fixed, paraffin embedded and sliced for immunohistochemical analysis or sorted by their eGFP expression for future analysis. The tested parameters were: proliferation (immunohistochemistry)/cell cycle (FACS), apoptosis (immunohistochemistry/FACS), cell count/adhesion/distribution around the placenta (cell sorter, visual observation and counting), matrix metalloproteinase activity (zymogram) and estrogen receptor (ER) expression (western blotting, immunohistochemistry). RESULTS: Reduced breast cancer cell numbers (45%↓, 48%↓ for MCF-7/T47D, respectively, P < 0.05) were observed near the placenta. The placenta elevated MCF-7 sub-G1 phase and modestly elevated apoptosis (3-17%↑ for T47D/MCF-7, respectively, P < 0.05). Our findings demonstrate breast cancer cell migration from the placenta as: (i) T47D/MCF-7 cells changed their morphology to that of motile cells; (ii) elevated MMPs activity was found in the co-culture; (iii) placental soluble factors detached breast cancer cells; and (4) the placenta reduced MCF-7/T47D cells' ER expression (a characteristic of motile cells). CONCLUSIONS: MCF-7/T47D cells are eliminated from the placental surroundings. Analyzing the causes of these phenomena may suggest biological pathways for this event and raise new therapeutic targets.

Tetraspanin-induced death of myeloma cell lines is autophagic and involves increased UPR signalling.

BACKGROUND: Multiple myeloma (MM) therapy is hindered by the interaction of the heterogeneous malignant plasma cells with their microenvironment and evolving drug resistance. We have previously shown that the membranal tetraspanins, CD81 and CD82, are under-expressed in MM cells and that their reintroduction causes massive non-apoptotic death. In this study, we aimed to characterise the tetraspanin-induced MM death. METHODS: Multiple myeloma cell lines were transiently transfected with eGFP-CD81N1/CD82N1 fusion proteins and assessed for death mode by flow cytometry (propidium iodide, ZVAD-fmk, 3MA), activation of unfolded protein response (UPR), and autophagy (immunoblot, RT-PCR). RESULTS: Cell death induced by CD81N1 and CD82N1 in MM cell lines was autophagic and involved endoplasmic reticulum (ER)-stress manifested by activation of UPR pathways, PERK (protein kinase-like ER kinase) and IRE1 (inositol-requiring 1). We also established the relative X-box binding protein 1 baseline expression levels in a panel of MM cell lines and their general dependence on autophagy for survival. Timeline of UPR cascades and cell fate supported our results. INTERPRETATION: This is the first publication implicating tetraspanins in UPR signalling pathways, autophagy, and autophagic death. Integration of our findings with published data highlights the unifying dependence of MM cells on ER-Golgi homoeostasis, and underscores the potential of tetraspanin complexes and ER-stress as leverage for MM therapy.

Complete rehabilitation of patients with bruxism by veneered and non-veneered zirconia restorations with an increased vertical dimension of occlusion: an observational case-series study.

PURPOSE: To assess the clinical performance of the complete rehabilitation of a series of patients with bruxism treated by teeth- and implant-supported veneered and non-veneered zirconia restorations with an increased vertical dimension of occlusion (VDO). METHODS: Ten patients with bruxism, aged 62-70 years, were treated with 108 veneered and 142 non-veneered zirconia restorations and attended the recall appointment. The mean observation period was 28.2 (±16.8) months. The patients were identified from records, and clinical details were retrieved from their files. In the recall appointment, the restorations were evaluated using modified California Dental Association (CDA) criteria. The periodontal probing depth, bleeding index, presence of caries and implant survival and success rate were recorded. RESULTS: No biological complications were recorded for any restorations. The success and survival rate of all implants was 100%. The overall mean survival and success rate of all restorations was 99.6%. In the veneered group, the predominant complication was minor veneer chipping (13.9%) on the incisal edge, which required only polishing (grade 1); in the non-veneered group, the predominant failure was open proximal contacts between the implant restoration and adjacent teeth (9%); only one implant restoration needed repair. One restoration was replaced due to a horizontal tooth fracture. CONCLUSIONS: Within the limitations of this study, we conclude that the survival and success rate of monolithic zirconia restorations installed in patients with bruxism was excellent, although the veneered zirconia restorations showed a high rate of minor veneer chipping, which required only polishing.

Safety, tolerability and efficacy of a novel self-use biodegradable device for management of obesity.

OBJECTIVE: Obesity is a major public health issue with significant impact on quality of life, morbidity and mortality rates. It is estimated that if the current trends continue, 18% of men and 21% of women worldwide will be obese by 2025. All the current therapies are not optimal due to limited efficacy or safety; thus, there is a need for additional devices for the treatment of obesity. This study aimed to examine the safety, tolerability, and efficacy of a biodegradable encapsulated Epitomee device for weight loss. The technology is based on absorbent pharmaceuticals polymers and bonding materials that self-expand in the stomach to create a pH-sensitive super absorbent gel structure for weight loss. METHODS: A prospective, 12-week twice daily use of the encapsulated device in patients with body mass index of 27-40 kg m-2. Efficacy endpoints were the percent total body weight loss (%TBWL), proportion of participants with 5% TBWL and changes in cardio-metabolic markers. Safety analysis included evaluation of adverse events, laboratory and endoscopic findings. RESULTS: Overall, 52 patients completed the study. TBWL per intension-to-treat analysis was 3.68 ± 3.07% (3.23 ± 2.69 kg) and 4.52 ± 2.97% (3.95 ± 2.57 kg) per protocol. No device serious adverse effects reported. The most common adverse events were headache (18.1%), viral infection (11.5%), abdominal discomfort (10.1%), bloating (7.9%), nausea and constipation (5% each) and flatulence (4.3%). Endoscopy in 26 patients revealed mild, asymptomatic gastric/duodenal erythema without erosions in five patients. CONCLUSIONS: Twelve weeks of Epitomee capsules treatment combined with lifestyle counselling resulted in 3.68-4.52% of TBWL. With continued research, the Epitomee capsules have considerable potential to become a non-invasive, safe and effective treatment option for weight loss.

The Role of heat shock protein 27 in extravillous trophoblast differentiation.

Trophoblast cells from placental explants differentiate in culture to extravillous trophoblast cells (EVT cells). During trophoblast differentiation heat-shock-protein-27 (HSP27) mRNA and multidrug-resistance-protein-5 (MRP5, transporter of cyclic nucleotides) expression are increased. HSP27 is a regulator of actin filaments structure and dynamic, has a role in cell differentiation and may affect NF-kB activity. In this study we aimed to assess HSP27 level in trophoblast cells and its correlation with motility and differentiation related processes [MMPs activity, nitric oxide (NO), inducible nitric oxide synthase (iNOS), proliferation and MRP5 levels]. We evaluated HSP27 expression in a first trimester human trophoblast explants model designed to assess EVT cells differentiation/migration with/without 6-mercaptopurine (6MP, an EVT inhibitor of migration). We found that HSP27 level is expressed in the nucleous and cytoplasm of non-proliferting villous-trophoblast cells (negative for Ki67) and in the cell periphery and cytoplasm of motile EVT cells. Moreover, 6MP decreased HSP27 nucleous expression that was associated with inhibited MMP2 activity and NO production. Also decreased iNOS expression and increased MRP5 mRNA levels were observed. In conclusion, HSP27 expression is modulated in concordance with migration dependent parameters in trophoblast cells.

Asbestos inhalation induces reactive nitrogen species and nitrotyrosine formation in the lungs and pleura of the rat.

To determine whether asbestos inhalation induces the formation of reactive nitrogen species, three groups of rats were exposed intermittently over 2 wk to either filtered room air (sham-exposed) or to chrysotile or crocidolite asbestos fibers. The rats were killed at 1 or 6 wk after exposure. At 1 wk, significantly greater numbers of alveolar and pleural macrophages from asbestos-exposed rats than from sham-exposed rats demonstrated inducible nitric oxide synthase protein immunoreactivity. Alveolar macrophages from asbestos-exposed rats also generated significantly greater nitrite formation than did macrophages from sham-exposed rats. Strong immunoreactivity for nitrotyrosine, a marker of peroxynitrite formation, was evident in lungs from chrysotile- and crocidolite-exposed rats at 1 and 6 wk. Staining was most evident at alveolar duct bifurcations and within bronchiolar epithelium, alveolar macrophages, and the visceral and parietal pleural mesothelium. Lungs from sham-exposed rats demonstrated minimal immunoreactivity for nitrotyrosine. Significantly greater quantities of nitrotyrosine were detected by ELISA in lung extracts from asbestos-exposed rats than from sham-exposed rats. These findings suggest that asbestos inhalation can induce inducible nitric oxide synthase activation and peroxynitrite formation in vivo, and provide evidence of a possible alternative mechanism of asbestos-induced injury to that thought to be induced by Fenton reactions.

Quantitation of nitrotyrosine levels in lung sections of patients and animals with acute lung injury.

Activated alveolar macrophages and epithelial type II cells release both nitric oxide and superoxide which react at near diffusion-limited rate (6.7 x 10(9) M-1s-1) to form peroxynitrite, a potent oxidant capable of damaging the alveolar epithelium and pulmonary surfactant. Peroxynitrite, but not nitric oxide or superoxide, readily nitrates phenolic rings including tyrosine. We quantified the presence of nitrotyrosine in the lungs of patients with the adult respiratory distress syndrome (ARDS) and in the lungs of rats exposed to hyperoxia (100% O2 for 60 h) using quantitative immunofluorescence. Fresh frozen or paraffin-embedded lung sections were incubated with a polyclonal antibody to nitrotyrosine, followed by goat anti-rabbit IgG coupled to rhodamine. Sections from patients with ARDS (n = 5), or from rats exposed to hyperoxia (n = 4), exhibited a twofold increase of specific binding over controls. This binding was blocked by the addition of an excess amount of nitrotyrosine and was absent when the nitrotyrosine antibody was replaced with nonimmune IgG. In additional experiments we demonstrated nitrotyrosine formation in rat lung sections incubated in vitro with peroxynitrite, but not nitric oxide or reactive oxygen species. These data suggest that toxic levels of peroxynitrite may be formed in the lungs of patients with acute lung injury.

Prognostic significance of supradiaphragmatic lymph nodes at initial presentation in patients with stage III high-grade serous ovarian cancer.

PURPOSE: To identify the optimal size threshold and to assess the prognostic significance of supradiaphragmatic lymph nodes at initial presentation of patients with high-grade serous ovarian cancer (HGSC). METHODS: This IRB-approved, HIPAA-compliant retrospective study included baseline pretreatment staging abdominal CTs of 88 women (mean age 62 years, SD 10.4, range 29-85) with FIGO stage III HGSC. Patients with stage IV disease were excluded due to worse prognosis and management guided by distant metastases. Two fellowship-trained radiologists independently reviewed abdominal CTs to record the presence of supradiaphragmatic nodes, abdominal lymphadenopathy, peritoneal carcinomatosis, and ovarian mass. Progression-free survival (PFS) and overall survival (OS) were recorded after median 79 months follow-up (IQR 58-115, range 13-144). The optimal short-axis size threshold for supradiaphragmatic lymphadenopathy was determined by correlating 3, 4, 5, 6, 7, and 10 mm thresholds with PFS and OS using Log-rank test. Prognostic significance of supradiaphragmatic lymphadenopathy was assessed using Cox proportional hazards models. RESULTS: There was good interobserver agreement for presence (κ = 0.65, 95%CI 0.51-0.79) and size (ICC = 0.77, 95%CI 0.66-0.86) of supradiaphragmatic nodes. 5 mm short-axis size threshold was associated with significantly shorter PFS (median 14 months, IQR 11-17 vs. 23 months, IQR 12-59; p = 0.02) and OS (median 44 months, IQR 27-69 vs. 65 months, IQR 45-96; p = 0.03). Total 38/88 (43%) patients had supradiaphragmatic lymphadenopathy. On Cox proportion hazards analysis, supradiaphragmatic lymphadenopathy was significantly associated with shorter PFS (p = 0.02; HR 1.81, 95%CI 1.11-2.96) and OS (p = 0.008; HR 2.11, 95%CI 1.21-3.65). CONCLUSION: In patients with stage III HGSC, supradiaphragmatic lymphadenopathy is associated with shorter PFS and OS. Further studies would help determine its implications on staging, decision regarding neoadjuvant therapy, and surgical technique.

Antibacterial properties of temporary filling materials.

The purpose of this study was to investigate the antibacterial properties of temporary fillings. The direct contact test (DCT) was used to evaluate the antibacterial properties of Revoltek LC, Tempit, Systemp inlay, and IRM. These were tested in contact with Streptococcus mutans and Enterococcus faecalis. The materials were examined immediately after setting, 1, 7, 14, and 30 days after aging in phosphate buffered saline (PBS). Statistical analysis included two-way ANOVA, one-way ANOVA, and Tukey multiple comparison. Systemp inlay, Tempit, and IRM exhibited antibacterial properties when in contact with S. mutans for at least 7 days, Tempit and IRM sustained this ability for at least 14 days. When in contact with E. faecalis Tempit and IRM were antibacterial immediately after setting, IRM sustained this ability for at least 1 day. Our study suggests that the difference in temporary filling materials may influence which microorganism will be able to invade the root canal system.