One-pot chemo- and photo-enzymatic linear cascade processes.

The combination of chemo- and photocatalyses with biocatalysis, which couples the flexible reactivity of the photo- and chemocatalysts with the highly selective and environmentally friendly nature of enzymes in one-pot linear cascades, represents a powerful tool in organic synthesis. However, the combination of photo-, chemo- and biocatalysts in one-pot is challenging because the optimal operating conditions of the involved catalyst types may be rather different, and the different stabilities of catalysts and their mutual deactivation are additional problems often encountered in one-pot cascade processes. This review explores a large number of transformations and approaches adopted for combining enzymes and chemo- and photocatalytic processes in a successful way to achieve valuable chemicals and valorisation of biomass. Moreover, the strategies for solving incompatibility issues in chemo-enzymatic reactions are analysed, introducing recent examples of the application of non-conventional solvents, enzyme-metal hybrid catalysts, and spatial compartmentalization strategies to implement chemo-enzymatic cascade processes.

Efficacy and safety of pembrolizumab in metastatic urothelial carcinoma: results from KEYNOTE-045 and KEYNOTE-052 after up to 5 years of follow-up.

BACKGROUND: Immune checkpoint inhibitors are a standard therapy in metastatic urothelial carcinoma (UC). Long-term follow-up is necessary to confirm durability of response and identify further safety concerns. PATIENTS AND METHODS: In KEYNOTE-045, patients with metastatic UC that progressed on platinum-containing chemotherapy were randomly assigned 1:1 to receive pembrolizumab or investigator's choice of paclitaxel, docetaxel, or vinflunine. Primary endpoints were progression-free survival per RECIST version 1.1 by blinded independent central review (BICR) and overall survival. In KEYNOTE-052, cisplatin-ineligible patients with metastatic UC received first-line pembrolizumab. The primary endpoint was objective response rate per RECIST version 1.1 by BICR. RESULTS: A total of 542 patients (pembrolizumab, n = 270; chemotherapy, n = 272) were randomly assigned in KEYNOTE-045. The median follow-up was 62.9 months (range 58.6-70.9 months; data cut-off 1 October 2020). At 48 months, overall survival rates were 16.7% for pembrolizumab and 10.1% for chemotherapy; progression-free survival rates were 9.5% and 2.7%, respectively. The median duration of response (DOR) was 29.7 months (range 1.6+ to 60.5+ months) for pembrolizumab and 4.4 months (range 1.4+ to 63.1+ months) for chemotherapy; 36-month DOR rates were 44.4% and 28.3%, respectively. A total of 370 patients were enrolled in KEYNOTE-052. The median follow-up was 56.3 months (range 51.2-65.3 months; data cut-off 26 September 2020). The confirmed objective response rate was 28.9% (95% confidence interval 24.3-33.8), and the median DOR was 33.4 months (range 1.4+ to 60.7+ months); the 36-month DOR rate was 44.8%. Most treatment-related adverse events for pembrolizumab in either study were grade 1 or 2 and manageable, which is consistent with prior reports. CONCLUSION: With ∼5 years of follow-up, pembrolizumab monotherapy continued to demonstrate durable efficacy with no new safety signals in patients with platinum-resistant metastatic UC and as first-line therapy in cisplatin-ineligible patients. CLINICAL TRIAL REGISTRY AND ID: With ClinicalTrials.gov NCT02256436 (KEYNOTE-045); https://clinicaltrials.gov/ct2/show/NCT02256436 and NCT02335424 (KEYNOTE-052); https://clinicaltrials.gov/ct2/show/NCT02335424.

Genomic characterisation of hormone receptor-positive breast cancer arising in very young women.

BACKGROUND: Very young premenopausal women diagnosed with hormone receptor-positive, human epidermal growth factor receptor 2-negative (HR+HER2-) early breast cancer (EBC) have higher rates of recurrence and death for reasons that remain largely unexplained. PATIENTS AND METHODS: Genomic sequencing was applied to HR+HER2- tumours from patients enrolled in the Suppression of Ovarian Function Trial (SOFT) to determine genomic drivers that are enriched in young premenopausal women. Genomic alterations were characterised using next-generation sequencing from a subset of 1276 patients (deep targeted sequencing, n = 1258; whole-exome sequencing in a young-age, case-control subsample, n = 82). We defined copy number (CN) subgroups and assessed for features suggestive of homologous recombination deficiency (HRD). Genomic alteration frequencies were compared between young premenopausal women (<40 years) and older premenopausal women (≥40 years), and assessed for associations with distant recurrence-free interval (DRFI) and overall survival (OS). RESULTS: Younger women (<40 years, n = 359) compared with older women (≥40 years, n = 917) had significantly higher frequencies of mutations in GATA3 (19% versus 16%) and CN amplifications (CNAs) (47% versus 26%), but significantly lower frequencies of mutations in PIK3CA (32% versus 47%), CDH1 (3% versus 9%), and MAP3K1 (7% versus 12%). Additionally, they had significantly higher frequencies of features suggestive of HRD (27% versus 21%) and a higher proportion of PIK3CA mutations with concurrent CNAs (23% versus 11%). Genomic features suggestive of HRD, PIK3CA mutations with CNAs, and CNAs were associated with significantly worse DRFI and OS compared with those without these features. These poor prognostic features were enriched in younger patients: present in 72% of patients aged <35 years, 54% aged 35-39 years, and 40% aged ≥40 years. Poor prognostic features [n = 584 (46%)] versus none [n = 692 (54%)] had an 8-year DRFI of 84% versus 94% and OS of 88% versus 96%. Younger women (<40 years) had the poorest outcomes: 8-year DRFI 74% versus 85% and OS 80% versus 93%, respectively. CONCLUSION: These results provide insights into genomic alterations that are enriched in young women with HR+HER2- EBC, provide rationale for genomic subgrouping, and highlight priority molecular targets for future clinical trials.

Prognostic factors for complications after loop ileostomy reversal.

BACKGROUND: Defunctioning ileostomy creation and closure are both associated with morbidity. There is little data available about complications after ileostomy closure. The aim of this study was to evaluate morbidity related to loop ileostomy closure (LIC) and to determine if patients with postoperative complications in primary surgery suffer from more postoperative complications during stoma closure. METHODS: This was a retrospective study on prospectively registered consecutive patients undergoing elective LIC in a single centre in Spain between April 2010 and December 2017. Baseline characteristics, postoperative complications after primary surgery and after stoma closure were recorded. Primary surgery included any colorectal resection, elective or urgent associated with a diverting loop ileostomy either as a protective stoma or rescue procedure. A logistic regression model was used to assess the effects of baseline variables and postoperative complications after primary surgery on the existence of postoperative complications related to LIC. RESULTS: Four hundred and twenty-eight patients (288 men, median age 64.5 years [IQR 55.1-72.3 years]) were included in the study, and 37.4%, developed complications after LIC. The most common was paralytic ileus. Only chronic kidney disease (OR 2.31; 95% CI 1.03-5.33, p = 0.043), existence of postoperative complications after primary surgery (OR 2.25; 95% CI 1.41-3.66, p = < 0.001) and ileostomy closure later than 10 months after primary surgery (OR 1.52; 95% CI 1.00-2.33, p = 0.049) were statistically significant in the multivariate analysis. CONCLUSIONS: Patients with chronic kidney disease, those who had any complication after primary surgery and those who had LIC > 10 months after primary surgery have a significantly higher risk of developing postoperative complications.

Height difference between the vestibular and palatal walls and palatal width: a cone beam computed tomography approach.

BACKGROUND: The objective of this study was to measure two parameters involved in tri-dimensional implant planning: the position of the buccal and palatal bone wall and the palatal thickness. METHODS: Cone beam computed tomography (CBCT) images (Planmeca ProMax 3D) of 403 teeth (208 upper teeth and 195 lower teeth) were obtained from 49 patients referred to the Dental School of Seville from January to December 2014. The height difference between the palatal and buccal walls was measured on the most coronal point of both walls. The thickness of the palatal wall was measured 2 mm from the most coronal point of the palatal wall. RESULTS: The mean values in the maxilla were 1.7 ± 0.9 mm for central and lateral incisors, 2.2 ± 1.7 mm for canines, 1.6 ± 0.9 mm for premolars and 1.9 ± 1.5 mm for molars. In the lower jaw, the mean values were 1.3 ± 0.8 mm for incisors, 1.7 ± 1.2 mm for canines, 2.3 ± 1.3 mm for premolars, and 2.6 ± 1.7 mm for molars. In the upper jaw, more than 55% of maxillary teeth (excluding second premolars and molars) presented mean height differences greater than 1 mm. In the mandible, more than 60% of incisors showed a buccal bone thickness of 1 mm from the apical to lingual aspect. All teeth except the second premolar presented a buccal wall located more than 1 mm more apically than the lingual bone wall. CONCLUSIONS: The buccal bone wall is located more apically (greater than 1 mm) than the palatal or lingual table in most of the cases assessed. The thickness of the palatal or lingual table is also less than 2 mm in the maxilla and mandible, except in the upper canines and premolars and the lower molars.

Randomized phase III KEYNOTE-045 trial of pembrolizumab versus paclitaxel, docetaxel, or vinflunine in recurrent advanced urothelial cancer: results of >2 years of follow-up.

BACKGROUND: Novel second-line treatments are needed for patients with advanced urothelial cancer (UC). Interim analysis of the phase III KEYNOTE-045 study showed a superior overall survival (OS) benefit of pembrolizumab, a programmed death 1 inhibitor, versus chemotherapy in patients with advanced UC that progressed on platinum-based chemotherapy. Here we report the long-term safety and efficacy outcomes of KEYNOTE-045. PATIENTS AND METHODS: Adult patients with histologically/cytologically confirmed UC whose disease progressed after first-line, platinum-containing chemotherapy were enrolled. Patients were randomly assigned 1 : 1 to receive pembrolizumab [200 mg every 3 weeks (Q3W)] or investigator's choice of paclitaxel (175 mg/m2 Q3W), docetaxel (75 mg/m2 Q3W), or vinflunine (320 mg/m2 Q3W). Primary end points were OS and progression-free survival (PFS) per Response Evaluation Criteria in Solid Tumors, version 1.1 (RECIST v1.1) by blinded independent central radiology review (BICR). A key secondary end point was objective response rate per RECIST v1.1 by BICR. RESULTS: A total of 542 patients were enrolled (pembrolizumab, n = 270; chemotherapy, n = 272). Median follow-up as of 26 October 2017 was 27.7 months. Median 1- and 2-year OS rates were higher with pembrolizumab (44.2% and 26.9%, respectively) than chemotherapy (29.8% and 14.3%, respectively). PFS rates did not differ between treatment arms; however, 1- and 2-year PFS rates were higher with pembrolizumab. The objective response rate was also higher with pembrolizumab (21.1% versus 11.0%). Median duration of response to pembrolizumab was not reached (range 1.6+ to 30.0+ months) versus chemotherapy (4.4 months; range 1.4+ to 29.9+ months). Pembrolizumab had lower rates of any grade (62.0% versus 90.6%) and grade ≥3 (16.5% versus 50.2%) treatment-related adverse events than chemotherapy. CONCLUSIONS: Long-term results (>2 years' follow-up) were consistent with those of previously reported analyses, demonstrating continued clinical benefit of pembrolizumab over chemotherapy for efficacy and safety for treatment of locally advanced/metastatic, platinum-refractory UC. TRIAL REGISTRATION: ClinicalTrials.gov: NCT02256436.

Crestal module design optimization of dental implants: finite element analysis and in vivo studies.

The aim of this study was to evaluate the biomechanical behavior of Bone Level dental implants with four different neck designs in contact with cortical bone. Numerical simulations were performed using a Finite Element Method (FEM) based-model. In order to verify the FEM model, the in silico results were compared with the results obtained from histological analysis performed in an in vivo study with New Zealand rabbits. FEM was performed using a computerized 3D model of Bone Level dental implants inserted in the lower jaw bone with an applied axial load of 100 N. The analysis was performed using four different implant neck designs: even surfaced, screwed, three-ring design and four-ring design. Interface are of bone growth was evaluated by analyzing the Bone-Implant-Contact (BIC) parameter obtained from in vivo histological process and analyzed by Scanning Electron Microscopy (SEM). Bone Level implants were inserted in the rabbit tibia, placing two implants per tibia. The BIC was evaluated after three and six weeks of implantation. FEM studies showed that the three-ring design presented lower values of stress distribution compared to the other studied designs. The lower levels of mechanical stress were then correlated with the in vivo studies, showing that the three-ring design presented the highest BIC value after 3 and 6 weeks of implantation. In silico and in vivo results both concluded that the implants with three-ring neck design presented the best biomechanical and histological behavior in terms of new bone formation, enhanced mechanical stability and optimum osseointegration.

Repeatability of the resonance frequency analysis values in implants with a new technology.

BACKGROUND: Assess the reliability (by means of reproducibility and repeatability) of the PenguinRFA system, analyse the ISQ values of different implant types and correlate the ISQ with the insertion torque during the placement of the implant. MATERIAL AND METHODS: 120 rough surface implants were placed in bovine bone (type II and III). The implants were divided into groups, according to its design. Once the implants were in place, the exact insertion torque was registered. Then, primary stability was measured by means of the resonance frequency analysis with the PenguinRFA and the Osstell ISQ devices. In each implant two transducers of each device were used. Three measurements were obtained with each transducer. RESULTS: The mean ISQ (implant stability quotient) of the whole sample is 67,70 ± 5,51. The Intraclass Correlation Coefficient (ICC) is 0,933 and 0,944 for transducers 1 and 2 respectively. The reproducibility is 0,906. The mean insertion torque is 24,54 ± 8,96N. The correlation between the ISQ and the insertion torque is 0,507 p<0,000 (MultiPeg 1) and 0,468 p<0,000 (MultiPeg 2) for bone type II and 0,533 p<0,801 (MultiPeg 1) and 0,193 p<0,140 (MultiPeg 2) for bone type III. CONCLUSIONS: The results of the present trial suggest that the PenguinRFA presents excellent reproducibility and repeatability, so it could be very useful in the monitoring of the stability of implants over time. Additionally, according to the results, the correlation between the IT and the RFA is low and there are no statistically significant differences in between implant types.

Unravelling the effect of macro and microscopic design of dental implants on osseointegration: a randomised clinical study in minipigs.

Several dental implants are commercially available and new prototype design are constantly being fabricated. Nevertheless, it is still unclear what parameters of the design affect most the osseointegration of dental implants. The purpose of this study is to assess the effects of the microscopic and macroscopic design of dental implants in the osseointegration by comparing three macroscopic designs (Straumann tissue level (STD), essential cone (ECD) and prototype design (PD)) and six surface treatments. A total of 96 implants were placed in 12 minipigs. The implant stability quotient (ISQ), was assessed at the time of implantation, as well as at 2, 4 and 8 weeks. Histomorphometric and statistical analyses were conducted at the different sacrifice times, being 2, 4 and 8 weeks, to analyse the bone to implant contact (BIC), the bone area density (BAT) and the density of bone outside the thread region (ROI). The macroscopic design results showed higher ISQ values for the ECD, whereas the histomorphometric analysis showed higher ossoeintegration values for the STD. Regarding the microscopic design, both Sandblasted plus acid etching (hydrochloric/sulphuric acid) in a nitrogen atmosphere (SLActive) and Shot-blasted or bombarded with alumina particles and posterior alkaline immersion and thermal treatment (ContacTi) showed superior results in terms of osseointegration and reduced the osseointegration times from 8 weeks to 4 weeks compared to the other analysed surfaces. In conclusion, each of the macroscopic and microscopic designs need to be taken into account when designing novel dental implants to enhance the osseointegration process.

Thickness of the buccal bone wall and root angulation in the maxilla and mandible: an approach to cone beam computed tomography.

BACKGROUND: The objective of this paper is to anatomically describe the bone morphology in the maxillary and mandibular tooth areas, which might help in planning post-extraction implants. METHODS: CBCT images (Planmeca ProMax 3D) of 403 teeth (208 upper teeth and 195 lower teeth) were obtained from 49 patients referred to the Dental School of Seville from January to December 2014. The thickness of the facial wall was measured at the crest, point A, 4 mm below, point B, and at the apex, point C. The second parameter was the angle formed between the dental axis and the axis of the basal bone. RESULTS: A total of 403 teeth were measured. In the maxilla, 89.4% of incisors, 93.94% of canines, 78% of premolars and 70.5% of molars had a buccal bone wall thickness less than the ideal 2 mm. In the mandible, 73.5% of incisors, 49% of canines, 64% of premolars and 53% of molars had < 1 mm buccal bone thickness as measured at point B. The mean angulation in the maxilla was 11.67 ± 6.37° for incisors, 16.88 ± 7.93° for canines, 13.93 ± 8.6° for premolars, and 9.89 ± 4.8° for molars. In the mandible, the mean values were 10.63 ± 8.76° for incisors, 10.98 ± 7.36° for canines, 10.54 ± 5.82° for premolars and 16.19 ± 11.22° for molars. CONCLUSIONS: The high incidence of a buccal wall thickness of less than 2 mm in over 80% of the assessed sites indicates the need for additional regeneration procedures, and several locations may also require custom abutments to solve the angulation problems for screw-retained crowns.

A randomized, double-blind, placebo-controlled phase II study of maintenance therapy with tasquinimod in patients with metastatic castration-resistant prostate cancer responsive to or stabilized during first-line docetaxel chemotherapy.

BACKGROUND: This phase II study was conducted to assess clinical efficacy of tasquinimod maintenance therapy in patients with metastatic castrate-resistant prostate cancer not progressing during first-line docetaxel-based therapy. PATIENTS AND METHODS: Patients were randomly assigned (1 : 1) to receive tasquinimod (0.25-1.0 mg/day orally) or placebo. The primary end point was radiologic progression-free survival (rPFS); secondary efficacy end points included: overall survival (OS); PFS on next-line therapy (PFS 2) and symptomatic PFS, assessed using the Brief Pain Inventory (BPI) questionnaire and analgesic use. Quality of life was measured by the Functional Assessment of Cancer Therapy-Prostate (FACT-P) questionnaire and by the EuroQol-5 Dimension Quality of Life Instrument (EQ-5D). Adverse events were recorded. RESULTS: A total of 219 patients were screened and 144 patients randomized. The median duration of treatment was 18.7 weeks (range 0.6-102.7 weeks) for the tasquinimod arm and 19.2 weeks (range 0.4-80.0 weeks) for the placebo arm. Median (90% CI) rPFS was 31.7 (24.3-53.7) and 22.7 (16.1-25.9) weeks in the tasquinimod and placebo arms, respectively [HR (90% CI) 0.6 (0.4-0.9), P = 0.0162]. The median OS was not reached because only 14 deaths occurred by the cut-off date. No statistically significant differences between treatment arms were noted for symptomatic PFS, PFS 2, BPI score, FACT-P score, or EQ-5D. The incidence of any treatment emergent adverse event (TEAE) was similar in the tasquinimod and placebo arms (97.2% versus 94.3%, respectively), whereas severe TEAEs (NCI-CTC Grade 3-5) incidence was higher in the tasquinimod group (50.7% versus 27.1%). CONCLUSIONS: Randomized trials testing new drugs as maintenance can be successfully conducted after chemotherapy in castrate-resistant prostate cancer. Maintenance tasquinimod therapy significantly reduced the risk of rPFS by 40%. CLINICALTRIALS: gov identifier NCT01732549.

Androgen receptor gene status in plasma DNA associates with worse outcome on enzalutamide or abiraterone for castration-resistant prostate cancer: a multi-institution correlative biomarker study.

BACKGROUND: There is an urgent need to identify biomarkers to guide personalized therapy in castration-resistant prostate cancer (CRPC). We aimed to clinically qualify androgen receptor (AR) gene status measurement in plasma DNA using multiplex droplet digital PCR (ddPCR) in pre- and post-chemotherapy CRPC. METHODS: We optimized ddPCR assays for AR copy number and mutations and retrospectively analyzed plasma DNA from patients recruited to one of the three biomarker protocols with prospectively collected clinical data. We evaluated associations between plasma AR and overall survival (OS) and progression-free survival (PFS) in 73 chemotherapy-naïve and 98 post-docetaxel CRPC patients treated with enzalutamide or abiraterone (Primary cohort) and 94 chemotherapy-naïve patients treated with enzalutamide (Secondary cohort; PREMIERE trial). RESULTS: In the primary cohort, AR gain was observed in 10 (14%) chemotherapy-naïve and 33 (34%) post-docetaxel patients and associated with worse OS [hazard ratio (HR), 3.98; 95% CI 1.74-9.10; P < 0.001 and HR 3.81; 95% CI 2.28-6.37; P < 0.001, respectively], PFS (HR 2.18; 95% CI 1.08-4.39; P = 0.03, and HR 1.95; 95% CI 1.23-3.11; P = 0.01, respectively) and rate of PSA decline ≥50% [odds ratio (OR), 4.7; 95% CI 1.17-19.17; P = 0.035 and OR, 5.0; 95% CI 1.70-14.91; P = 0.003, respectively]. AR mutations [2105T>A (p.L702H) and 2632A>G (p.T878A)] were observed in eight (11%) post-docetaxel but no chemotherapy-naïve abiraterone-treated patients and were also associated with worse OS (HR 3.26; 95% CI 1.47-not reached; P = 0.004). There was no interaction between AR and docetaxel status (P = 0.83 for OS, P = 0.99 for PFS). In the PREMIERE trial, 11 patients (12%) with AR gain had worse PSA-PFS (sPFS) (HR 4.33; 95% CI 1.94-9.68; P < 0.001), radiographic-PFS (rPFS) (HR 8.06; 95% CI 3.26-19.93; P < 0.001) and OS (HR 11.08; 95% CI 2.16-56.95; P = 0.004). Plasma AR was an independent predictor of outcome on multivariable analyses in both cohorts. CONCLUSION: Plasma AR status assessment using ddPCR identifies CRPC with worse outcome to enzalutamide or abiraterone. Prospective evaluation of treatment decisions based on plasma AR is now required. CLINICAL TRIAL NUMBER: NCT02288936 (PREMIERE trial).

Clinical significance and outcome of Aeromonas spp. infections among 204 adult patients.

The objectives of this investigation were to analyze the clinical patterns, risk groups, prognostic factors, and mortality of infections caused by Aeromonas spp. This was a retrospective study of adult patients with Aeromonas spp. isolates attended at the Hospital del Mar in Barcelona, Spain, between January 2006 and December 2012. Epidemiological data, antimicrobial susceptibility, clinical patterns, underlying illnesses, type of infection, admission to the intensive care unit (ICU), number of episodes, coinfection, antimicrobial therapy, and evolution were analyzed. A total of 221 clinical samples from 204 patients were positive for Aeromonas spp. The mean age of the patients was 67.6 years. The main clinical form of presentation was gastrointestinal (78.4%). Malignancy was the main risk group in 69 (33.8%) patients, and 48 (23.5%) were previously healthy. Twenty-one patients (10.3%) were admitted to the ICU. Infections were acquired in the hospital in 52.5% of the patients, and 28.9% were polymicrobial. The overall mortality (after 1 year of follow-up from the first positive culture) was 26.5%. Univariate analysis identified an association between increased mortality and the following variables: age ≥80 years, hospitalization, admission to the ICU, malignancy, extraintestinal infection, and appropriate antimicrobial therapy. In the multivariate analysis, age ≥80 years [odds ratio (OR), 4.37 [95% confidence interval (CI), 1.68-11.35; p = 0.002]], admission to the ICU (OR, 6.59 [95% CI, 2.17-19.99; p = 0.001]), and malignancy (OR, 3.62 [95% CI, 1.32-9.90; p = 0.012]) were significantly associated with mortality. Aeromonas infections are mainly gastrointestinal. The 1-year follow-up mortality rate was high. Old age (age ≥80 years), admission to the ICU, and malignancy were identified as independent risk factors for mortality.

New advances in genitourinary cancer: evidence gathered in 2014.

This review provides updated information published in 2014 regarding advances and major achievements in genitourinary cancer. Sections include the best in prostate cancer, renal cancer, bladder cancer, and germ cell tumors. In the field of prostate cancer, data related to treatment approach of hormone-sensitive disease, castrate-resistant prostate cancer, mechanisms of resistance, new drugs, and molecular research are presented. In relation to renal cancer, relevant aspects in the treatment of advanced renal cell carcinoma, immunotherapy, and molecular research, including angiogenesis and von Hippel-Lindau gene, molecular biology of non-clear cell histologies, and epigenetics of clear renal cell cancer are described. New strategies in the management of muscle-invasive localized bladder cancer and metastatic disease are reported as well as salient findings of biomolecular research in urothelial cancer. Some approaches intended to improve outcomes in poor prognosis patients with metastatic germ cell cancer are also reported. Results of clinical trials in these areas are discussed.

Salicylic acid is required for Mi-1-mediated resistance of tomato to whitefly Bemisia tabaci, but not for basal defense to this insect pest.

Plant defense to pests or pathogens involves global changes in gene expression mediated by multiple signaling pathways. A role for the salicylic acid (SA) signaling pathway in Mi-1-mediated resistance of tomato (Solanum lycopersicum) to aphids was previously identified and its implication in the resistance to root-knot nematodes is controversial, but the importance of SA in basal and Mi-1-mediated resistance of tomato to whitefly Bemisia tabaci had not been determined. SA levels were measured before and after B. tabaci infestation in susceptible and resistant Mi-1-containing tomatoes, and in plants with the NahG bacterial transgene. Tomato plants of the same genotypes were also screened with B. tabaci (MEAM1 and MED species, before known as B and Q biotypes, respectively). The SA content in all tomato genotypes transiently increased after infestation with B. tabaci albeit at variable levels. Whitefly fecundity or infestation rates on susceptible Moneymaker were not significantly affected by the expression of NahG gene, but the Mi-1-mediated resistance to B. tabaci was lost in VFN NahG plants. Results indicated that whiteflies induce both SA and jasmonic acid accumulation in tomato. However, SA has no role in basal defense of tomato against B. tabaci. In contrast, SA is an important component of the Mi-1-mediated resistance to B. tabaci in tomato.

Expression of endogenous retroviruses is negatively regulated by the pluripotency marker Rex1/Zfp42.

Rex1/Zfp42 is a Yy1-related zinc-finger protein whose expression is frequently used to identify pluripotent stem cells. We show that depletion of Rex1 levels notably affected self-renewal of mouse embryonic stem (ES) cells in clonal assays, in the absence of evident differences in expression of marker genes for pluripotency or differentiation. By contrast, marked differences in expression of several endogenous retroviral elements (ERVs) were evident upon Rex1 depletion. We demonstrate association of REX1 to specific elements in chromatin-immunoprecipitation assays, most strongly to muERV-L and to a lower extent to IAP and musD elements. Rex1 regulates muERV-L expression in vivo, as we show altered levels upon transient gain-and-loss of Rex1 function in pre-implantation embryos. We also find REX1 can associate with the lysine-demethylase LSD1/KDM1A, suggesting they act in concert. Similar to REX1 binding to retrotransposable elements (REs) in ES cells, we also detected binding of the REX1 related proteins YY1 and YY2 to REs, although the binding preferences of the two proteins were slightly different. Altogether, we show that Rex1 regulates ERV expression in mouse ES cells and during pre-implantation development and suggest that Rex1 and its relatives have evolved as regulators of endogenous retroviral transcription.

Implant-abutment connections: influence of the design on the microgap and their fatigue and fracture behavior of dental implants.

Microgap between implant and abutment can produce biological and mechanical problems such as peri-implantitis and/or fatigue failures. The aim of this study was to evaluate microgap size and fatigue behavior of external and internal connections. In both systems the torque to tighten the abutment screw of single crown abutments was 45 Ncm. Fifty implants for each connection type were studied. One subgroup (n = 5) was used by the observation and evaluation of the microgap, other (n = 5) was tested for fracture strength and the other (n = 40) was subjected to dynamic loading. The internal connection presents a lower microgap than the external ones. From fatigue results, the external hexagon interface showed superior result compared to the internal hexagon interfaces. The tolerances in the internal connections are better and favour the fatigue behavior but this factor alone is not sufficient to improve the fatigue response in relation to the external connections when the screw is subjected at the same torque. The external system presents a higher value of the area than the internal and it produces a better load distribution. Microgaps and mechanical properties are very important for the long-term behavior of the dental implants and these aspects should be known by the implantologists.