Validation of the QLQ-EN24 instrument for the assessment of health-related quality of life for women with endometrial cancer in México.

PURPOSE: The impact of disease activity or treatments on health-related quality of life (HRQL) is crucial in Oncology, but adequate instruments for this assessment are scarce. Our aim is to validate the Mexican-Spanish version of the QLQ-EN24 questionnaire to evaluate HRQL in women with endometrial cancer (EC). METHODS: This is a prospective study of Mexican women with EC, attending a single cancer centre, who responded the QLQ-C30 and QLQ-EN24 instruments; usual psychometric analysis were performed as well as the association of HRQL scales and relevant clinical data. Correlation analysis was performed with the Spearman's method, reliability analysis with the Cronbach's alpha, known-group comparisons with the Kruskal-Wallis test, and survival analysis with the Kaplan-Meier method and Log-rank test. RESULTS: One hundred and eighty-nine women with EC were assessed. Most functional scales reported high values, and most symptom scales, low. Questionnaire compliance rates were high and internal consistency tests demonstrated adequate convergent and divergent validity. Cronbach's α coefficients of the five multi-item scales the QLQ-EN24 instruments were from 0.659 to 0.887. Scales of the QLQ-C30 and QLQ-EN24 instruments distinguished among clinically distinct groups of patients, particularly based on serum albumin levels. The Urological symptoms, Gastrointestinal symptoms, Body image, Pelvic pain and Taste change scales were significantly associated with OS. CONCLUSION: The Mexican-Spanish version of the QLQ-EN24 questionnaire is reliable and valid for the assessment of HRQL in patients with EC and can be broadly used in multi-national clinical trials. However, conclusions derived from scales evaluating sexual function should be handled carefully.

Depressed Supercooling Point and Increased Glycerol Concentration in Overwintering Adult Tiger Beetles (Cicindelida).

BACKGROUND: Tiger beetles are a widely distributed group including species that may be exposed to sub-freezing temperature overwinter. Despite being well studied, little is known about tiger beetle cold tolerance. OBJECTIVE: We investigated seasonal changes in cold hardiness of two northerly distributed tiger beetle species (Cicindela repanda and Cicindela limbalis). MATERIALS AND METHODS: We monitored the supercooling point (SCP), glycerol concentration, and hemolymph osmolality of adult tiger beetles during a 3.5-month acclimation to winter. RESULTS: SCP decreased during winter acclimation for C. repanda, but not for C. limbalis. Both species modestly increased glycerol concentration, and C. repanda increased hemolymph osmolality by 38%. CONCLUSION: This initial investigation into the cold-hardiness of adult tiger beetles suggests that they are capable of lowering their SCP as winter approaches, which may help them survive sub-freezing winter temperatures. Further assessment of their chill and freeze tolerance and of their overwintering conditions in the field is needed to better understand their winter physiology.

Effective use of mesenchymal stem cells in human skin substitutes generated by tissue engineering.

Mesenchymal stem cells (MSCs) can differentiate toward epithelial cells and may be used as an alternative source for generation of heterotypical artificial human skin substitutes, thus, enhancing their development and translation potential to the clinic. The present study aimed at comparing four types of heterotypical human bioengineered skin generated using MSCs as an alternative epithelial cell source. Adipose-tissue-derived stem cells (ADSCs), dental pulp stem cells (DPSCs), Wharton's jelly stem cells (WJSCs) and bone marrow stem cells (BMSCs) were used for epidermal regeneration on top of dermal skin substitutes. Heterotypic human skin substitutes were evaluated before and after implantation in immune-deficient athymic mice for 30 d. Histological and genetic studies were performed to evaluate extracellular matrix synthesis, epidermal differentiation and human leukocyte antigen (HLA) molecule expression. The four cell types differentiated into keratinocytes, as shown by the expression of cytokeratin 10 and filaggrin 30 d post-grafting; also, they induced dermal fibroblasts responsible for the synthesis of extracellular fibrillar and non-fibrillar components, in a similar way among each other. WJSCs and BMSCs showed higher expression of cytokeratin 10 and filaggrin, suggesting these cells were more prone to epidermal regeneration. The absence of HLA molecules, even when the epithelial layer was differentiated, supports the future clinical use of these substitutes - especially ADSCs, DPSCs and WJSCs - with low rejection risk. MSCs allowed the generation of bioengineered human skin substitutes with potential clinical usefulness. According to their epidermal differentiation potential and lack of HLA antigens, WJSCs should preferentially be used.

Endotoxin levels after chemomechanical preparation of root canals with sodium hypochlorite or chlorhexidine: a systematic review of clinical trials and meta-analysis.

AIM: The aim of this systematic review was to answer the following question: in patients with primary endodontic infection, is there a statistically significant difference in the endotoxin levels after chemomechanical preparation with sodium hypochlorite (NaOCl) or chlorhexidine (CHX)? METHODOLOGY: A protocol was prepared and registered on PROSPERO (CRD42017069996). Four electronic databases (MEDLINE via PubMeb, Scopus, Web of Science and Cochrane Library) were searched from their start dates to 1 March 2017 using strict inclusion and exclusion criteria and reviewed following PRISMA (Preferred Reporting Items for Systematic reviews and Meta-Analyses) guidelines. Only clinical trials (randomized and nonrandomized) that compared the effectiveness of NaOCl and CHX to reduce endotoxins during chemomechanical preparation of teeth with primary endodontic infection were included. Two reviewers independently assessed the eligibility for inclusion, extracted data and assessed the quality using the risk of bias tool. RESULTS: From 712 articles that resulted from the initial search, 37 studies were included for full-text appraisal; four studies met the inclusion criteria for quantitative synthesis. A single meta-analysis was performed to compare the endotoxin levels before and after chemomechanical preparation with NaOCl or CHX. The forest plot of lipopolysaccharide (LPS) levels indicated that the data were heterogeneous [I2  = 63.9%; Tau2  = 574.5 (P = 0.04)]. The use of NaOCl and CHX during chemomechanical preparation significantly reduced the LPS levels compared to the initial ones. CONCLUSIONS: Chemomechanical canal preparation with both NaOCl and CHX reduced the endotoxin levels compared to the initial ones found in primary endodontic infections. When NaOCl was used during chemomechanical preparation, endotoxins levels were lower than those obtained after chemomechanical preparation with CHX.

Wharton's jelly-derived mesenchymal cells as a new source for the generation of microtissues for tissue engineering applications.

Microtissues (MT) are currently considered as a promising alternative for the fabrication of natural, 3D biomimetic functional units for the construction of bio-artificial substitutes by tissue engineering (TE). The aim of this study was to evaluate the possibility of generating mesenchymal cell-based MT using human umbilical cord Wharton's jelly stromal cells (WJSC-MT). MT were generated using agarose microchips and evaluated ex vivo during 28 days. Fibroblasts MT (FIB-MT) were used as control. Morphometry, cell viability and metabolism, MT-formation process and ECM synthesis were assessed by phase-contrast microscopy, functional biochemical assays, and histological analyses. Morphometry revealed a time-course compaction process in both MT, but WJSC-MT resulted to be larger than FIB-MT in all days analyzed. Cell viability and functionality evaluation demonstrated that both MT were composed by viable and metabolically active cells, especially the WJSC during 4-21 days ex vivo. Histology showed that WJSC acquired a peripheral pattern and synthesized an extracellular matrix-rich core over the time, what differed from the homogeneous pattern observed in FIB-MT. This study demonstrates the possibility of using WJSC to create MT containing viable and functional cells and abundant extracellular matrix. We hypothesize that WJSC-MT could be a promising alternative in TE protocols. However, future cell differentiation and in vivo studies are still needed to demonstrate the potential usefulness of WJSC-MT in regenerative medicine.

Periodontal education and assessment in the undergraduate dental curriculum-A questionnaire-based survey in European countries.

OBJECTIVES: This survey aimed to evaluate whether periodontal education and assessment in undergraduate dental curricula amongst the member countries of the European Federation of Periodontology (EFP) follow the competency-based curricular guidelines and recommendations developed by the Association for Dental Education in Europe. MATERIALS AND METHODS: A multiple-choice questionnaire was emailed to 244 dental institutes amongst the 24 EFP member countries between November 2014 and July 2015. RESULTS: Data were received from 16 (66.7%) EFP member countries. Out of 117 responding dental institutes, 76 (64.95%) were included as valid responders. In most of the institutes (86.3%), a minimum set of competencies in periodontology was taken into account when constructing their dental education programmes. Out of 76 responders, 98.1% included lecture-based, 74.1% case-based and 57.1% problem-based teaching in their periodontal curricula, whilst a minority (15.9%) also used other methods. A similar pattern was also seen in the time allocation for these four educational methods, that is, the highest proportion (51.8%) was dedicated to lecture-based teaching and only a small proportion (5.7%) to other methods. Periodontal competencies and skills were most frequently assessed by clinical grading on clinic, multiple-choice examination (written examination) and oral examination, whereas competency tests and self-assessment were rarely used. Only in 11 (14.5%) cases, access flap procedures were performed by students. CONCLUSION: Great diversity in teaching methodology amongst the surveyed schools was demonstrated, and thus, to harmonise undergraduate periodontal education and assessment across Europe, a minimum set of recommendations could be developed and disseminated by the EFP.

Clinical efficacy of EDTA ultrasonic activation in the reduction of endotoxins and cultivable bacteria.

AIM: This clinical study was conducted to investigate the influence of 17% ethylenediaminetetraacetic acid (EDTA) ultrasonic activation after chemomechanical preparation (CMP) on eliminating/reducing oral bacterial lipopolysaccharides (known as endotoxins) and cultivable bacteria in teeth with pulp necrosis and apical periodontitis. METHODOLOGY: Samples were taken from 24 root canals at several clinical periods: S1 - before CMP; S2 - after CMP; S3 - after EDTA: G1 - with ultrasonic activation (n = 12) and G2 - without ultrasonic activation (n = 12). Root canals were instrumented using Mtwo rotary files. Culture techniques were used to determine the number of colony-forming units (CFU). Limulus amebocyte lysate (LAL) was used to measure endotoxin levels. Friedman's and Wilcoxon signed-rank tests were used to compare the amount of bacteria and endotoxin levels in each period (P < 0.05). RESULTS: Endotoxins and cultivable bacteria were recovered in 100% of the initial samples (S1). CMP was effective in reducing endotoxins and bacterial load (all with P < 0.05). Higher values of endotoxin reduction were achieved with EDTA ultrasonic activation [G1, 0.02 EU mL-1 (range 0.01-0.75)] compared with the no activation group [G2, 1.13 EU mL-1 (range 0.01-8.34)] (P < 0.05). Regarding bacterial reduction, no statistically significant difference was found in S3, regardless of the group (G1, G2, P > 0.05). CONCLUSIONS: Chemomechanical preparation was effective in reducing bacteria and endotoxins, but could not completely eliminate them. The ultrasonic activation of EDTA was effective in further reducing endotoxin levels in the root canals of teeth with pulp necrosis and apical periodontitis.

Influence of apical enlargement and complementary canal preparation with the Self-Adjusting File on endotoxin reduction in retreatment cases.

AIM: To compare the effectiveness of large apical preparations and complementary canal preparation with the Self-Adjusting File (SAF) in removing endotoxins from the root canal of teeth with apical periodontitis. METHODOLOGY: Ten single-rooted and single-canaled teeth with post-treatment apical periodontitis were selected. Endotoxin samples were taken after removal of the root filling (S1), after chemomechanical preparation (CMP) using 2.5% NaOCl and an R25 file (S2), after CMP using 2.5% NaOCl and an R40 file (S3) and after complementary CMP using the SAF system (S4). Limulus amebocyte lysate (LAL) was used to measure endotoxin levels. The Friedman and Wilcoxon tests were used to compare endotoxin levels at each clinical intervention (P < 0.05). RESULTS: After root filling removal, endotoxin was detected in 100% of the root canals (S1, 4.84 EU mL-1 ). CMP with the R25 file was able to significantly reduce endotoxin levels (P < 0.05). Increased levels of endotoxin removal were achieved by apical preparation with the R40 file (P < 0.05). Complementary CMP with SAF did not significantly reduce endotoxin levels (P > 0.05) following the use of the R40 instrument. CONCLUSIONS: Apical enlargement protocols were effective in significantly reducing endotoxin levels. Complementary preparation with the SAF system failed to eliminate residual endotoxin contents beyond those obtained with the R40 instrument.

Evaluation of new chlorhexidine- and cetylpyridinium chloride-based mouthrinse formulations adjunctive to scaling and root planing: pilot study.

OBJECTIVE: To compare the effect of two newly formulated chlorhexidine (CHX) and cetylpyridinium chloride (CPC) mouthrinses after scaling and root planing (SRP) in terms of clinical, microbiological, patient-based variables and adverse events, with a positive control with the same active components, already marketed and tested. METHODS: A pilot, randomized clinical trial, double-blind, parallel design with 1-month follow-up was conducted. Chronic periodontitis patients requiring non-surgical periodontal therapy were enrolled and randomly assigned to: (i) SRP and test-1 (new reformulation: 0.12% CHX and 0.05% CPC); (ii) SRP and test-2 (new formulation: 0.03% CHX and 0.05% CPC); or (iii) SRP and positive control (commercial product: 0.12% CHX and 0.05% CPC). All variables were evaluated at baseline and 1 month after SRP. Quantitative variables were compared by means of anova or Kruskal-Wallis test and qualitative variables by chi-square or McNemar tests. RESULTS: Thirty patients (10 per group) were included. After 1 month, there were significant differences among groups in plaque levels (P = 0.016) as test-1 showed less sites with plaque than test-2 (31.15% [standard error-SE 2.21%] versus 49.39% [SE 4.60%), respectively). No significant differences were found for global patient perception of the product or in adverse effects. Test groups showed better results in levels and proportions (P = 0.022) of Capnocytophaga spp. CONCLUSIONS: Within the limitations of this pilot study, it can be concluded that the newly formulated 0.12% CHX and 0.05% CPC mouthrinse showed larger plaque level reductions, without showing more adverse effects, when compared to the other two mouthrinses, after SRP.

Characterization of A. actinomycetemcomitans strains in subgingival samples from periodontitis subjects in Morocco.

OBJECTIVE: Aggregatibacter actinomycetemcomitans, specially its highly leucotoxic strain (JP2 clone), represents an etiological factor for the onset and progression of aggressive types of periodontitis. The aims of this investigation were to investigate the most relevant periodontal pathogens in the subgingival microbiota of periodontitis patients from Morocco and to describe the clinical and microbiological characteristics of subjects positive for A. actinomycetemcomitans, including serotype, leukotoxin gene, and operon of the cytolethal distending toxin (cdt) distribution. MATERIAL AND METHODS: In consecutive Moroccan subjects diagnosed of periodontitis, subgingival samples were taken and processed by culture. From the positive samples for A. actinomycetemcomitans, one to three isolates were subcultured and characterized by means of polymerase chain reaction (PCR), assessing their specific serotype distribution, the variation in the sequences of the leukotoxin gene, and the operon of the cdt. RESULTS: Twenty-one (35.6 %) out of 59 periodontitis patients harbored A. actinomycetemcomitans. These patients demonstrated statistically significant deeper pockets (p = 0.035) and higher proportions of P. micra (p = 0.045) than did the negative group. The 39 studied isolates were serotype "b"; in 16 out of 17 patients, there was mono-colonization with this serotype. Five isolates, from two patients, presented the 530-bp deletion in the leukotoxin's promoter region. Thirty-two isolates (78 % of the strains) were cdt-positive. CONCLUSION: A. actinomycetemcomitans was frequently found (35.6 %) in our sample. All strains were serotype "b," and most (78 %) were also cdt-positive. The JP2 strain type was only detected in 12.2 % of the strains. CLINICAL RELEVANCE: A. actinomycetemcomitans can be frequently found in Morocco. This fact can influence the therapeutic approach of this type of patients.

Comparison of the detection of periodontal pathogens in bacteraemia after tooth brushing by culture and molecular techniques.

BACKGROUND: The prevalence and amounts of periodontal pathogens detected in bacteraemia samples after tooth brushing-induced by means of four diagnostic technique, three based on culture and one in a molecular-based technique, have been compared in this study. MATERIAL AND METHODS: Blood samples were collected from thirty-six subjects with different periodontal status (17 were healthy, 10 with gingivitis and 9 with periodontitis) at baseline and 2 minutes after tooth brushing. Each sample was analyzed by three culture-based methods [direct anaerobic culturing (DAC), hemo-culture (BACTEC), and lysis-centrifugation (LC)] and one molecular-based technique [quantitative polymerase chain reaction (qPCR)]. With culture any bacterial isolate was detected and quantified, while with qPCR only Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans were detected and quantified. Descriptive analyses, ANOVA and Chi-squared tests, were performed. RESULTS: Neither BACTEC nor qPCR detected any type of bacteria in the blood samples. Only LC (2.7%) and DAC (8.3%) detected bacteraemia, although not in the same patients. Fusobacterium nucleatum was the most frequently detected bacterial species. CONCLUSIONS: The disparity in the results when the same samples were analyzed with four different microbiological detection methods highlights the need for a proper validation of the methodology to detect periodontal pathogens in bacteraemia samples, mainly when the presence of periodontal pathogens in blood samples after tooth brushing was very seldom.

Radiographic features of the development of the anterior tibial tuberosity. / Características radiológicas del desarrollo de la tuberosidad tibial anterior.

OBJECTIVE: Few studies have evaluated the radiologic characteristics of the development of the anterior tibial tuberosity. This study aimed to evaluate the radiologic characteristics of the anterior tibial tuberosity in a pediatric population broken down into age groups. MATERIAL AND METHODS: We assessed 210 plain-film X-rays of the knee from patients aged from 10 to 17 years, divided into groups according to age and sex, for the presence of ossification of the anterior tibial tuberosity, the distance between the anterior tibial tuberosity and the metaphysis, and fusion with the epiphysis. RESULTS: At 10 years of age, the anterior tibial tuberosity was ossified in 50% of the girls but in only 25% of the boys. In all the girls, the anterior tibial tuberosity was ossified at 11 years, fusion of the anterior tibial tuberosity with the epiphysis had started at 12 years, and fusion was complete by 17 years. In boys, the process is delayed by one year compared to girls. A single center of ossification was found in all cases. CONCLUSION: The ossification of the anterior tibial tuberosity starts distally, then the proximal part fuses with the rest of the epiphysis, and finally the distal part fuses with the tibia. The results of this study help enable a better analysis of the anterior tibial tuberosity in cases of knee pain.

Root canal content from primary endodontic infection and upregulation of gelatinases in fibroblast cells.

AIM: To investigate endotoxin levels from primary endodontic infections before and after chemomechanical preparation (CMP) and to determine their antigenicity against 3T3 fibroblasts through gelatinolytic activity of matrix metalloproteinases (MMPs). METHODOLOGY: Twenty-four root canals with primary endodontic infection and apical periodontitis were selected. Samples were collected using paper points before (S1) and after chemomechanical preparation (CMP) (S2). The limulus amebocyte lysate assay was used for endotoxin measurement. Fibroblasts were stimulated with root canal contents for 24 h. Supernatants of cell cultures stimulated with root canal contents were collected after 24 h to determine the levels of MMP-2 and MMP-9 gelatinolytic activity using the zymography technique. Friedman and Wilcoxon tests were used to compare the amount of endotoxin before (S1) and after CMP (S2) (P < 0.05). Data obtained from gelatinolytic activity were analysed using anova and Tukey's tests (P < 0.05). RESULTS: Endotoxin was recovered in 100% of the samples. There was a significant reduction in endotoxin levels after CMP (P < 0.05). A correlation was found between the levels of endotoxins and MMP-2 expression (P < 0.05). Root canal contents of initial samples (S1) induced significantly greater MMP-2 expression by fibroblasts when compared to S2 and the nonstimulated group (P < 0.05). No gelatinolytic activity of MMP-9 was observed in S1, S2 and control group. CONCLUSIONS: Root canal contents from primary endodontic infections had gelatinolytic activity for MMP-2. Moreover, CMP was effective in reducing endotoxin levels and their antigenicity against fibroblasts on gelatinolytic activity.

Characterization and application of a flow system for in vitro multispecies oral biofilm formation.

BACKGROUND AND OBJECTIVE: Bacteria in the oral cavity grow in the form of biofilms; these structures are subject to constant saliva or gingival crevicular fluid flow conditions. The aims of this study were: (i) to develop and to characterize an in-vitro biofilm model with oral bacteria growing under flow and shear conditions; and (ii) to demonstrate the usefulness of the model for evaluating the activity of three antiplaque agents. MATERIAL AND METHODS: We used a bioreactor to grow the oral bacteria Streptococcus oralis, Actinomyces naeslundii, Veillonella parvula, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis under planktonic conditions. Biofilms were established using a modified Robbins device on hydroxyapatite (HAP) discs. Three- to 7-d-old biofilms were analysed using culture methods, scanning electron microscopy, Live/Dead staining and fluorescence in-situ hybridization (confocal laser scanning microscopy). Finally, we assessed the antimicrobial activity of three mouthrinses [0.12% chlorhexidine (CHX), 0.12% chlorhexidine and sodium fluoride (CHX+NaF) and 0.12% chlorhexidine and 0.05% cetylpyridinium chloride (CHX+CPC)] using a planktonic test (short interval-killing test) and in our 4-d biofilm model. RESULTS: The viable cell counts showed that each species was consistently found in the biofilms throughout the study. The architecture and cell distribution were similar to those described for biofilms in situ, with the exception of a thin layer of living cells that was found close to the HAP. The effectiveness test of the mouthwashes demonstrated that cells in biofilms showed more tolerance compared with planktonic cells. Moreover, it was observed that in 4-d biofilm formed in vitro, CHX+CPC caused significantly higher mortality compared with CHX (p = 0.003) and CHX+NaF (p < 0.001). CONCLUSION: Our results suggest that we have a highly reproducible system for multispecies oral biofilm formation and that it is a useful tool for assessing antibacterial molecules before their clinical evaluation. It also has great potential to be used in basic research on supragingival and subgingival biofilms.

Quantitative real-time PCR combined with propidium monoazide for the selective quantification of viable periodontal pathogens in an in vitro subgingival biofilm model.

BACKGROUND AND OBJECTIVES: Differentiation of live and dead cells is an important challenge when using molecular diagnosis for microbial identification. This is particularly relevant when bacteria have been exposed to antimicrobial agents. The objective of this study was to test a method using quantitative real-time polymerase chain reaction (qPCR) combined with propidium monoazide (PMA), developed for the selective quantification of viable P. gingivalis, A. actinomycetemcomitans, F. nucleatum and total bacteria in an in vitro biofilm model after antimicrobial treatment. MATERIAL AND METHODS: PMA-qPCR method was tested in an in vitro biofilm model, using isopropyl alcohol as the antimicrobial agent. Matured biofilms were exposed for 1, 5, 10 and 30 min to isopropyl alcohol by immersion. Biofilms were disrupted and PMA added (final concentration of 100 µm). After DNA isolation, qPCR was carried out using specific primers and probes for the target bacteria. The differentiation of live and dead cells was tested by analysis of variance. RESULTS: When PMA was used in the presence of viable target bacterial cells, no statistically significant inhibition of qPCR amplification was detected (p > 0.05 in all cases). Conversely, after immersion in isopropyl alcohol of the biofilm, PMA resulted in a significant total reduction of qPCR amplification of about 4 log10 . P. gingivalis showed a vitality reduction in the biofilm of 3 log10 , while A. actinomycetemcomitans and F. nucleatum showed a 2 log10 reduction. CONCLUSION: These results demonstrate the efficiency of PMA for differentiating viable and dead P. gingivalis, A. actinomycetemcomitans and F. nucleatum cells, as well as total bacteria, in an in vitro biofilm model, after being exposed to an antimicrobial agent. Hence, this PMA-qPCR method may be useful for studying the effect of antimicrobial agents aimed at oral biofilms.

First Report of High Boscalid Resistance in Botrytis cinerea Associated with the H272L Mutation in Grapevine in Chile.

Gray mold (Botrytis cinerea Pers.) is a major disease of grapevine (Vitis vinifera L.) worldwide. Integrated control strategies, including canopy management and fungicide treatments, are needed to control gray mold. Chemical control relies on the use of single mode of action fungicides. The aim of this research was to study the sensitivity of B. cinerea to boscalid, which is a single mode of action fungicide of the succinate dehydrogenase inhibitor (SDHI) fungicide group. Fifty isolates were obtained in 2012 to 2013 from commercial vineyards in central Chile. Vineyards had received two boscalid applications at least for four consecutive years. Briefly, the percent mycelial growth inhibition (MGI) was determined on minimal medium (MM) (2) plus 50 µg m-1 of boscalid (Cantus 50 WP, BASF Chile). Each isolate was tested in triplicate, obtaining 2% highly resistant (HR, MGI ≤25%), 32% moderately resistant (MR, MGI 26 to 50%), 64% low resistant (LR, MGI 51 to 80%), and 2% sensitive (S, MGI ≥81%) phenotypes. Nine isolates were arbitrary selected and compared for MGI on MM plus 50 µg ml-1 of boscalid (1) and conidial germination inhibition (CGI) on yeast extract-bacto peptone-Na acetate (YBA) plus 5 µg ml-1 of boscalid (2,3). Isolates previously determined to be S and HR had the same phenotype for both MGI and CGI. However, all of the MR and LR isolates, determined based on the MGI tests, were identified as S isolates in the CGI tests. Using primer-introduced restriction analysis (PIRA)-PCR (4), the SdhB mutations were detected only in the HR isolate. The amplifications were performed with H272L-fw/H272-rev and were digested by the enzyme BglII, yielding 35- and 85-bp fragments and confirming a mutation at codon 272 (H272L) in the HR phenotype. The efficacy of the label-rate (0.4 g liter-1) boscalid in controlling gray mold was determined on 'Granny Smith' apples. The apples were surface-disinfested (75% ethanol, 30 s), wounded with a sterile syringe, and inoculated with a mycelium plug (5 mm in diameter) or 20 µl of a conidial suspension (106 conidia/ml) of one HR, MR, and S isolate. The inoculum was placed on the wounded sites after boscalid application. Apples were incubated for 7 days at 21°C. Each test had four replicates and the experiment was conducted three times. Boscalid slightly controlled (<6.7% efficacy) gray mold on the apples that were inoculated with mycelium or conidia of the HR phenotype isolate, while the sensitive isolate was highly controlled (>95% efficacy), and the MR isolate was moderately controlled (27 to 34% efficacy). These results demonstrate that mycelium or conidia assays using MM + 50 µg ml-1 boscalid or YBA+5 µg ml-1 boscalid consistently detected HR isolates. The S isolates detected using MGI were also S according with the CGI tests. The presence of the boscalid HR strains of B. cinerea associated with the H272L mutation in grapevine in Chile is reported for the first time in this study. This finding suggests that resistance to boscalid needs to be considered in the design of gray mold control strategies in commercial grapevine orchards. References: (1) D. Fernandez-Ortuño et al Plant Dis. 96:1198, 2012. (2) M.-J. Hu et al. J. Phytopathol. 159:616, 2011. (3) Y. K. Kim and C. L. Xiao. Plant Dis. 94:604, 2010. (4) T. Veloukas et al. Plant Dis. 95:1302, 2011.

Site percolation threshold of composite square lattices and its agroecology applications.

We analyze the percolation threshold of square lattices comprising a combination of sites with regular and extended neighborhoods. We found that the percolation threshold of these composed systems smoothly decreases with the fraction of sites with extended neighbors. This behavior can be well-fitted by a Tsallis q-Exponential function. We found a relation between the fitting parameters and the differences in the gyration radius among neighborhoods. We also compared the percolation threshold with the critical susceptibility of nearest and next-to-nearest neighbor monoculture plantations vulnerable to the spread of phytopathogen. Notably, the critical susceptibility in monoculture plantations can be described as a linear combination of two composite systems. These results allow the refinement of mathematical models of phytopathogen propagation in agroecology. In turn, this improvement facilitates the implementation of more efficient computational simulations of agricultural epidemiology that are instrumental in testing and formulating control strategies.

Pulse quality analysis on soliton pulse compression and soliton self-frequency shift in a hollow-core photonic bandgap fiber.

A numerical investigation of low-order soliton evolution in a proposed seven-cell hollow-core photonic bandgap fiber is reported. In the numerical simulation, we analyze the pulse quality evolution in soliton pulse compression and soliton self-frequency shift in three fiber structures with different cross-section sizes. In the simulation, we consider unchirped soliton pulses (of 400 fs) at the wavelength of 1060 nm. Our numerical results show that the seven-cell hollow-core photonic crystal fiber, with a cross-section size reduction of 2%, promotes the pulse quality on the soliton pulse compression and soliton self-frequency shift. For an input soliton pulse of order 3 (which corresponds to an energy of 1.69 µJ), the pulse gets compressed with a factor of up to 5.5 and a quality factor of 0.73, in a distance of 12 cm. It also experiences a soliton-self frequency shift of up to 28 nm, in a propagation length of 6 m, with a pulse shape quality of ≈ 0.80.

Association between periodontal status and pre-term and/or low-birth weight in Spain: clinical and microbiological parameters.

BACKGROUND AND OBJECTIVE: Studies performed over the last 15 years have suggested that periodontal diseases may be associated with adverse pregnancy outcomes. However, this association has not been found in all populations. The aim of this study was to evaluate whether periodontal status and the presence of specific periodontal pathogens may influence the incidence of adverse pregnancy outcomes. MATERIAL AND METHODS: Pregnant women were clinically examined before 26th week of gestation, and divided in two groups: non-periodontitis and periodontitis. Microbial samples were obtained in the periodontitis group and processed by anaerobic culturing. After delivery, data on the pregnancy outcome were taken; mother's socio-demographic and risk factors were obtained at inclusion. Simple and multiple regression analysis were performed. RESULTS: One hundred and seventy women were included in the study (116 non-periodontitis and 54 with periodontitis). The incidence of preterm (PTB) and low-birth weight (LBW) was 2.94% and 3.53%, respectively. Periodontal status did not show any association with adverse pregnancy outcomes. The presence of Eikenella corrodens was significantly related to PTB (p = 0.022) and the presence Capnocytophaga spp. was related to LBW (p = 0.008). The multivariate analyses showed a significant association between PTB and newborn weight and counts of E. corrodens. Maternal health and counts of E. corrodens were significantly associated with PTB or LBW. CONCLUSION: The clinical periodontal condition was not associated with adverse pregnancy outcomes in a Spanish Caucasian population with medium-high educational level. The presence and counts of E. corrodens and the presence of Capnocytophaga spp. showed a significant association with PTB and LBW, respectively, in the bivariate and/or multivariate model.

Analysis of viable vs. dead Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis using selective quantitative real-time PCR with propidium monoazide.

BACKGROUND AND OBJECTIVES: One of the major disadvantages of DNA-based microbial diagnostics is their inability to differentiate DNA between viable and dead microorganisms, which could be important when studying etiologically relevant pathogens. The aim of this investigation was to optimize a method for the selective detection and quantification of only viable Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis cells by combining quantitative real-time polymerase chain reaction (qPCR) and propidium monoazide (PMA). MATERIAL AND METHODS: Three different concentrations of PMA (10, 50 or 100 µm) were added to suspensions of 10(6) (CFU)/mL of viable/dead A. actinomycetemcomitans and P. gingivalis cells. After DNA isolation, qPCR was carried out using specific primers and probes for the tested bacteria. PMA was further tested with different mixtures containing varying ratios of viable and dead cells. The efficacy of PMA to detect viable/dead cells was tested by analysis of variance. RESULTS: For these specific bacterial pathogens, 100 µm PMA resulted in a significant reduction of qPCR amplification with dead cells (10(6) CFU/mL), while with viable cells no significant inhibition was detected. PMA was also effective in detecting selectively viable cells by qPCR detection, when mixtures of varying ratios of viable and dead bacteria were used. CONCLUSIONS: This study demonstrated the efficiency of PMA for differentiating viable and dead A. actinomycetemcomitans and P. gingivalis cells. This method of PMA-qPCR may be useful for monitoring new antimicrobial strategies and for assessing the pathogenic potential of A. actinomycetemcomitans and P. gingivalis in different oral conditions when using molecular diagnostic methods.