Research has shown that a factor in a victim’s forgiveness of an offender is the victim’s ability to make more positive, or at least less negative, attributions of the offender’s behavior and that perspective-taking can be a factor in facilitating that process. Self-enhancing humor has been found to be positively correlated with perspective-taking empathy and aggressive humor found to be negatively correlated with perspective-taking empathy. Therefore it was predicted that self-enhancing humor would be positively correlated with forgiveness and aggressive humor negatively correlated with forgiveness. The Humor Styles Questionnaire, the Absence of Negative and Presence of Positive subscales of the Forgiveness Scale, and the Forgiveness Likelihood Scale were administered to 112 college undergraduates. Self-enhancing humor was significantly and positively correlated with all of the forgiveness measures, aggressive humor and self-defeating humor were significantly and negatively correlated with some of the forgiveness measures and affiliative humor was not significantly correlated with any of the forgiveness measures. The results were interpreted in terms of previous findings for humor styles, perspective-taking empathy, depression, self-esteem and anxiety. Future research involving the extent to which other personality variables, such as perspective-taking empathy, mediate the relationship between self-enhancing humor and forgiveness was suggested. Forgiveness, defined as the victim letting go of negative affect (e.g. hostility), negative cognitions (e.g. thoughts of revenge), and negative behavior  toward the offender  has beenBesides marital quality and the offender’s remorse, perspective-taking could be another variable that promotes positive attributions of the offender’s behavior and thus forgiveness. There is considerable research on the relationship between humor and perspective-taking, in particular in regards to reducing stress by taking a more positive perspective of one’s situation. A study by The purpose of this study was to investigate the relationship between the four humor scales and forgiveness. It was predicted that self-enhancing humor would be positively correlated with forgiveness due to its association with perspective-taking empathy, which in turn is positively correlated with forgiveness . Since tM = 22.82, SD = 7.96).The participants were 112 students (39 men and 73 women) at a community college in the Midwestern United States. They ranged in age from 17 to 57  and a Presence of Positive subscale with five items reflecting positive feelings, thoughts and behaviors towards the offender (e.g. “I have been able to let go of my anger toward the person who wronged me”) that asks participants how they have responded to a person who has wronged or mistreated them. Each item has five response options in a Likert-type format (1 = Strongly Disagree to 5 = Strongly Agree). Validity was established with significant correlations for both subscales in the expected direction with measures of religiousness, hope, trait and state anger, spiritual and religious well-being and the Enright Forgiveness Inventory. The test-retest reliability was .76 for both the Absence of Negative and the Presence of Positive subscales and .80 for the total scale . The Cror = .41, p < .001) and the Presence of Positive subscale  and Absence of Negative subscale  of the Forgiveness Scale. The Cronbach alpha for the Forgiveness Likelihood Scale for the current sample was .85.The Forgiveness Likelihood Scale has ten items  that asks the participants how likely they would be to forgive someone in an imagined scenario. Each item has five response options in Likert-type format . Validity was supported with significant correlations in the expected direction with religiousness, religious well-being, trait anger and the Enright Forgiveness Inventory. Each of the two subscales of the Forgiveness Scale was significantly correlated with the Forgiveness Likelihood Scale. The test-retest reliability of the Forgiveness Likelihood Scale was .81. . In the The Humor Styles Questionnaire has 32 items. Each scale, affiliative humor (e.g. “I laugh and joke a lot with my closest friends”), self-enhancing humor , aggressive humor , and self-defeating humor (e.g. “I will often get carried away in putting myself down if it makes my family or friends laugh”) has eight items. Each item has seven response options in Likert-type format . The convergent validity for the affiliative humor scale was established by significant positive correlations with Extraversion on the NEO PI-R and Miller Social Intimacy Scale. Discriminant validity for the self-enhancing scale was indicated by a significant negative correlation with scores for Neuroticism on the NEO PI-R, and convergent validity was indicated with significant positive correlations with the Humor Coping subscale of the Coping Orientation to Problems Experienced Scale and the Coping Humor Scale. Convergent validity for the aggressive humor scale was estimated by a significant positive correlation with scores on the Cook-Medley Hostility Scale. Discriminant validity for the self-defeating humor scale was supported by significant negative correlations with the Rosenberg Self-Esteem Scale and with the Index of Self-Esteem . The CroProduct-Moment correlations between each of the four humor styles with the Forgiveness Likelihood Scale, the total Forgiveness Scale, and the Absence of Negative and the Presence of Positive subscales of the Forgiveness Scale were computed see  and 2. Ir = .34, p < .001), both the Absence of Negative  and Presence of Positive  subscales of the Forgiveness Scale and The Forgiveness Likelihood Scale  were all significant. None of the correlations between affiliative humor and the forgiveness measures, albeit positive, were significant. The correlations between aggressive humor and the total Forgiveness Scale  and the Presence of Positive subscale of the Forgiveness Scale  were significant, but those between aggressive humor and the Forgiveness Likelihood Scale and the Absence of Negative subscale of the Forgiveness Scale were not. The negative correlations between self-defeating humor and the total Forgiveness Scale , the Absence of Negative subscale of the Forgiveness Scale  and Presence of Positive subscale of the Forgiveness Scale , but not that between self-defeating humor and the Forgiveness Likelihood Scale, were significant.The positive correlations between self-enhancing humor and the total Forgiveness Scale (p < .001), both the Absence of Negative  and Presence of Positive  subscales of the Forgiveness Scale and the Forgiveness Likelihood Scale . Affiliative humor did not significantly predict any of the forgiveness scales or subscales. Aggressive humor significantly negatively predicted the Forgiveness Likelihood Scale  and the Presence of Positive subscale of the Forgiveness Scale , but did not significantly predict the total Forgiveness Scale or the Absence of Negative subscale of the Forgiveness Scale. Self-defeating humor significantly negatively predicted the total Forgiveness Scale  and the Absence of Negative subscale  of the Forgiveness Scale, but did not significantly predict the Forgiveness Likelihood Scale or the Presence of Positive subscale of the Forgiveness Scale.Self-enhancing humor positively and significantly predicted the total Forgiveness Scale  and was significantly negatively correlated with three of them, it did not significantly predict nor was significantly correlated with the Forgiveness Likelihood Scale. In fact, self-defeating humor was positively correlated with and positively predicted the Forgiveness Likelihood Scale, albeit not significantly. This could be due to the fact that the items on the Forgiveness Likelihood Scale represent hypothetical situations while the Forgiveness Scale involves items derived from actual situations in the participant’s life. Those high in self-defeating humor tend to be psychologically vulnerable, given the positive correlation between self-defeating humor with depression and anxiety and negative correlation with self-esteem  and posiThe current study has several limitations. First of all it is correlational in nature. It is not clear whether the humor style affects forgiveness, forgiveness affects the humor style, or some third variable is affecting both. Secondly, the participant sample study was somewhat homogenous in terms of age. Perhaps the relationship between the humor styles and forgiveness could change as a person matures. Thirdly the sample size could be larger. Finally, the Cronbach alpha for affiliative humor (.65) is relatively low. Replicating this study with a sample size at least twice as large and with a higher alpha for affiliative humor could strengthen the conclusions drawn from it.There are several directions that future research on humor styles and forgiveness could take based on the limitations of the current study. Studies could be done teasing out the causal relationships between humor styles and forgiveness. Researchers could possibly train participants to use self-enhancing humor better and then see if their forgiveness scores are higher than those of a control group. Another study could use the method developed by Longitudinal studies could also be conducted to see if the relationship between humor styles and forgiveness changes as one gets older, studying a group of participants as they age from adolescence to adulthood. If this would be too time-consuming or impractical, a cross-sectional study with participants of different ages could still illuminate how the relationship between humor styles and forgiveness changes as one grows older.Future research also is needed to determine how much of the variance for forgiveness is accounted by variables that are correlated with self-enhancing humor.


The environmental impacts of land use vary regionally. Differences in geomorphology, climate, landscape structure, and biotope inventories are regarded as the main causes of this variation. We present a methodological approach for identifying regional responses in land use type to large-scale changes and the implications for the provision of habitat for farmland birds. The methodological innovations of this approach are (i) the coupling of impact assessments with economic models, (ii) the linking of cropping techniques at the plot scale with the regional distribution of land use, and (iii) the integration of statistical or monitoring data on recent states. This approach allows for the regional differentiation of farmers’ responses to changing external conditions and for matching the ecological impacts of land use changes with regional environmental sensitivities. An exemplary scenario analysis was applied for a case study of an area in Germany, assessing the impacts of increased irrigation and the promotion of energy cropping on farmland birds, evaluated as a core indicator for farmland biodiversity. The potential effects on farmland birds were analyzed based on the intrinsic habitat values of the crops and cropping techniques. The results revealed that the strongest decrease in habitat availability for farmland birds occurred in regions with medium-to-low agricultural yields. As a result of the limited cropping alternatives, the increase in maize production was highest in marginal regions for both examined scenarios. Maize production replaced many crops with good-to-medium habitat suitability for birds. The declines in habitat quality were strongest in regions that are not in focus for conservation efforts for farmland birds. The decline in biological diversity in European agricultural landscapes is well documented in regional and Europe-wide studies Defra . ProjectAt the national level, biodiversity strategies and action plans ) have been developed in 180 countries across the world CBD . Most ofBirds on farmlands have exhibited the largest declines compared to birds that prefer other habitats Defra . Shifts Meaningful strategies to halt biodiversity losses must be based on cause–effect analyses and knowledge of the interdependencies of driving forces. Recent agricultural developments in Europe, such as energy cropping, are connected to increased input efficiency and thus support higher levels of specialized production systems. The term “intensification” is thus used to describe the impact of several interconnected mechanisms  in the majority of studies  directive and 27 bird reserves according to the European directive ) ) MUGV . DespiteA)) MUGV . While tA)) MUGV .Fig. 1LoThe landscapes in Brandenburg have been shaped by glacial and post-glacial processes. The soil conditions and landscape structure are typical for the northern portion of central continental Europe. The broad range in soil fertility and highly heterogeneous distribution determine the regional agricultural land use capabilities. The agricultural production is limited by annual precipitation  and by the predominance of loamy sand or sandy loam soils with limited yield potentials. The agricultural area in the study region has been classified according to the soil yield potential and cropping options , and the continued fostering of renewable energy production through national subsidy programs. As a baseline for the assessments, we used a business as usual (BAU) scenario. The BAU scenario extrapolated the ongoing progress in plant breeding programs for increases in the yields of crops into the year 2025. The BAU scenario was contrasted with an irrigation scenario and an energy scenario. The detailed assumptions for all the three scenarios are summarized in Table The initial working hypothesis is that the habitat quality of farmland is strongly influenced by land use changes, here, by changes in the composition and distribution of crops and their related management practices. The spatial distribution of crops is an economic decision that depends on land quality, available resources, preferences of the decision-making farmers, frame conditions such as the agro-environmental programs, and market conditions . The spatial variation in responses to changing frame conditions may lead to varying ecological sensitivities.Analysis and selection of future trends to be considered (with stakeholder involvement)Translation of the trends into framework conditions for different scenarios (scenario definition)Allocation of land use options under the scenario conditions Identification of relevant crop management activitiesAssessment of the impacts of relevant crop management activities on farmland birdsAggregation of crop management activities into one farmland bird indicator per land use option and soil rating classApplication to land use patterns obtained from the economic scenario analysisAssessments of land management impacts for the particular scenarios based on farmland bird indicatorsComparison with data on natural inventories Evaluation of the resulting field bird indicesThe main methodological steps are the following:Figure ©. The assumptions and constraints were (i) constant livestock fodder requirements, (ii) complete use of manure in the cropping systems, (iii) constant levels of contract-based cropping systems, and (iv) adherence to crop rotation restrictions. On this basis, the model maximized the total net margin by district by allocating all portions of the area of every soil category to the best-performing cropping practices when taking the restrictions into account.The economic analysis assesses the impacts of internal and external land use drivers on cropping practices and crop distributions based on scenario assumptions and site characteristics. We used a simplified linear programming optimization model for “region farms” to represent the economic decision-making of farmers in accordance to Rounsevell et al. . Each reYIELD estimation based on STATistics (YIELDSTAT) , Little owl (Athene noctua), Black tailed godwit (Limosa limosa)) are related to specific forest structures or to grasslands and were excluded. We distinguished two groups of birds: (i) birds that breed on arable land (skylark (Alauda arvensis), corn bunting , lapwing (Vanellus vanellus), and winchat (Saxicola rubetra)) and (ii) birds that only feed on arable land (red-backed shrike (Lanius collurio), the yellowhammer (Emberiza citrinella), and the woodlark (Lullula arborea)).We chose the “indicator for species diversity and landscape quality” as used within the German National Biodiversity Strategy BMU . The indThe land use impact module addresses the changes in crop species and crop management on the habitat availability or suitability for wildlife species that use arable farmland as their main habitat. This module is based on the basic assumption that crops serve as “foundation species”  and every bird species (N = 7) and then transferred to the soil rating classes by calculating area-weighted averages for the crop composition of the farm units of a particular region. Species behaviors and habitat preferences were parameterized , which is the reference layer since the beginning of the bird monitoring.Since many of the regional data on environmental sensitivities or on regional planning in Germany refer to a geomorphological classification scheme for Germany  to the modeling approach (Janssen and van Ittersum The results of the land use scenarios clearly reveal the uneven distribution of the cropping options and cropping alternatives across the study area. In relation to the agricultural and economic options, the farmers act divergently to changes in external economic conditions, depending on the respective soil quality class of their land. The results of the economic farm module indicate that the set of available cropping options increases together with the yield potential. The less fertile arable regions are less resilient against land use changes. Bioenergy cropping has often been suggested as an option to increase crop diversity, even on marginal land. Gehrels et al.  have repFurther research is required to quantify the amount and type of ecological compensation areas which will be needed outside of the agricultural fields to compensate for negative impacts of changes in crop management. We suggest that this will vary depending on landscape contexts.


Little is known about changes within the mitochondrial (mt) genome during tumor progression in general and during initiation and progression of neuroblastoma (NB) in particular. Whole exome sequencing of corresponding healthy tissue, primary tumor and relapsed tumor from 16 patients with NB revealed that most NB harbor tumor-specific mitochondrial variants. In relapsed tumors, the status of mt variants changed in parallel to the status of nuclear variants, as shown by increased number and spatio-temporal differences of tumor-specific variants, and by a concomitant decrease of germline variants. As mt variants are present in most NB patients, change during relapse and have a higher copy number compared to nuclear variants, they represent a promising new source of biomarkers for monitoring and phylogenetic analysis of NB. The mt genome differs from the nuclear genome by its high copy number, small size, intronless genomic structure, different mode of replication, exposure to oxygen radicals and increased DNA damage , 2. WhilNB is the most common extracranial solid tumor of childhood, with a poor prognosis in advanced stages . The nucTo check whether mt DNA alterations are present in NB, we analyzed mt sequences extracted from whole exome sequencing data of corresponding normal tissue, primary (diagnostic) tumors and relapsed tumors of 16 NB patients  that we Next, we wanted to know whether the status of mt variants changes during the course of NB. Indeed, in 14 out of 16 patients, changes from tumor at diagnosis to relapse were seen . In partAn unique opportunity to investigate the phylogeny of NB relapse arose in patient #12, who suffered a series of five relapses, the third (R3) and forth (R4) of which occurred concurrently at different sites Figure . A phyloFinally, we assessed how the mutational spectrum of the mt genome evolves during progression of NB in patients. To this end, the mutational signature of primary and relapsed tumors were obtained comparing mt variant motif spectra during tumor initiation and relapse. The spectrum of mutations at initial disease was consistent with replication-induced but not with reactive oxygen species-induced DNA damage  and did In this study we show for the first time that the majority of NB harbors mt DNA variants and that relapsing NB displays enhanced evolutional changes. The changes of the mt variants in NB at relapse, as manifested by increased number and spatio-temporal differences of tumor-specific variants, and by the concomitant decrease of germline variants, most likely has been caused by heterogeneous genetic drift of mt variants, given that selection of mt variants has only been observed with nonsense mutations . CumulatWe previously investigated the same set of corresponding primary and relapsed NB for the dynamics of nuclear mutations . DespiteWhile mitochondrial phylogenetic analysis of relapses in patient #12 was congruent with the corresponding nuclear analysis  in delinMutation spectra differed between the mt and nuclear genomes, suggesting different mutational mechanisms. This may be explained by the differences in genomic structure, replication, DNA damage and DNA repair between the two genomes. The stable mt mutation spectrum of primary and relapsed tumors suggests that similar mutational mechanisms act upon the mt genome at the time of diagnosis and relapse. In contrast, the changing nuclear mutation spectrum  proposesTaken together, we have determined the presence and change of mt DNA variants during neuroblastomagenesis. Notwithstanding whether mt variants have functional consequences and represent actionable targets in NB, they are a promising source of biomarkers for monitoring and phylogenetic analysis of NB.Corresponding blood and tumor tissue at diagnosis and at relapse were collected from 16 NB patients diagnosed and treated in participating study centers of the German neuroblastoma trials NB 97 and 2004. All participants provided informed consent. Detailed patient characteristics can be found in These methods are described in detail elsewhere . Brieflyhttp://www.mitomap.org) and mtDB: Human Mitochondrial Genome Database (http://www.mtdb.igp.uu.se/).Whole exome sequencing data was filtered for mitochondrial sequences and screened for mt variants using the MitoSeek tool  and the Tumor-specific mt variants. Corresponding primary and relapse tumor samples were first compared to normal tissue to identify tumor-specific mt variants. The paired difference between primary tumor and relapse for every patient was taken to assess the change in number of tumor-specific mt variants across all patient samples. To obtain a single parameter expressing the changes of all mt variants, the mean of the paired differences between primary tumor and relapse was calculated. The null hypothesis of the mean of the paired differences being equal to 0 when there is no increment or decrement of variants across sample types was tested. To this end, Monte Carlo simulation with 10,000 iterations was performed using randomly generated uniform numbers within the range of the observed values. The exact p-value was calculated.Germline mt variants. A linear regression model was fitted for every patient and the slope of the fitted line was taken as the coefficient that describes the trend of changes of the number of variants between normal tissue, primary tumor and relapse. To obtain a single parameter expressing the changes of all mt variants across all patients and samples, the mean of these coefficients was calculated. The null hypothesis, i.e. the mean of these coefficients being 0 when the number of variants in normal tissue, primary tumor and relapse are randomly distributed, was tested by Monte Carlo simulation and the exact p-value was calculated.All mt variants that passed the frequency filtering threshold  were used for the reconstruction of the phylogenetic tree in MEGA6 . The evoWe adapted a method previously described , 16 to oThe GraphPad Prism 6.01 software  was used.


Over the past seven years, the Department of Pediatric Dentistry at New York University College of Dentistry (NYUCD) and the Advanced Practice: Pediatrics and the Pediatric Nurse Practitioner (PNP) program at New York University College of Nursing (NYUCN) have engaged in a program of formal educational activities with the specific goals of advancing interprofessional education, evidence-based practice, and interprofessional strategies to improve the oral-systemic health of infants and young children. Mentoring interprofessional students in all health care professions to collaboratively assess, analyze, and care-manage patients demands that faculty reflect on current practices and determine ways to enhance the curriculum to include evidence-based scholarly activities, opportunities for interprofessional education and practice, and interprofessional socialization. Through the processes of interprofessional education and practice, the pediatric nursing and dental faculty identified interprofessional performance and affective oral health core competencies for all dental and pediatric primary care providers. Students demonstrated achievement of interprofessional core competencies, after completing the interprofessional educational clinical practice activities at Head Start programs that included interprofessional evidence-based collaborative practice, case analyses, and presentations with scholarly discussions that explored ways to improve the oral health of diverse pediatric populations. The goal of improving the oral health of all children begins with interprofessional education that lays the foundations for interprofessional practice. In 2005, New York University created a unique partnership between the College of Dentistry (NYUCD) and the College of Nursing (NYUCN) to foster interprofessional education through collaborative curriculum designs and practice environments with the goal of creating men and women of science whose education and practices were embedded in the principles of evidence-based dentistry, nursing, and medicine. Seven years ago, the pediatric faculty in their respective departments of dentistry and graduate pediatric nursing collaborated to provide interprofessional educational experiences in practice, research, and scholarship for second-year dental students, pediatric dental residents, and PNP students. The purpose of these collaborations was to provide formal interprofessional opportunities for these students to collaboratively examine the tenets of interprofessional practice and the evidence-based, culturally sensitive approaches to oral-systemic health care for culturally diverse, underserved children under 5 years old who are at high risk for dental caries. Two major goals were established by the nursing and dental faculty: (1) identify interprofessional pediatric oral health core competencies to enable the students to collaboratively plan interventions that reduce the incidence of early childhood caries (ECC); (2) promote evidence-based practice through interprofessional case analysis, discussions, and presentations. The purpose of this paper is to describe the design, implementation, and evaluation of dental and nursing interprofessional educational activities designed to accomplish these goals. Framework for Action on Interprofessional Education and Collaborative Practice [IPE is defined as “the collaborative process by which teams of health professions develop curricula and courses to coordinate and plan practical experiences jointly and team-teach groups of interdisciplinary [interprofessional] health professional students to provide holistic care through the life span” , p. 213. Team-based health care is the provision of health services to individuals, families, and/or their communities by at least two health providers who work collaboratively with patients and their caregivers—to the extent preferred by each patient—to accomplish shared goals within and across settings to achieve coordinated, high-quality care , p. 5.The Interprofessional Education Collaborative (IPEC) was launched in 2012 with six founding member organizations leading the initiative to educate health care professionals as teams with a focus on patient-centered care . FoundinThe principles of the IPEC can be readily applied to oral-systemic health care for the pediatric population. Oral-systemic health is defined as the relationship between oral health and systemic or whole body health , 7. The Early childhood caries (ECC) is defined as the presence of one or more decayed, missing, or filled tooth surfaces in any primary tooth in a child under 71 months of age . ECC isThe authors designed a nonexperimental descriptive study to observe and describe student interactions during interprofessional educational activities involving pediatric dental students, residents, and PNP students. Educational activities included interprofessional education, clinical practice, and professional socialization for this population of health care providers. The interprofessional clinical activities were designed to identify the evidence-based pediatric oral health core competencies for pediatric dental and PNP students.Study participants for the interprofessional oral health educational clinical experiences included Pediatric Nurse Practitioner students in their second year of study in a three-year part-time program, second-year dental students, and pediatric dental residents.Pediatric nurse practitioners (PNPs) are currently educated at the master level to provide primary care (PNP-PC) or acute care (PNP-AC) to children and adolescents. By 2015, educational preparation for nurse practitioners will be transitioning to doctoral education as entry into advanced practice .Dental students at NYUCD enter a 4-year program that includes didactic and clinical educational experiences. In their second year of study, the dental students participate in a program called ICE, which is an acronym for the “Initial Clinical Experience.” In addition, the ICE program was also their first organized interprofessional educational experience.The pediatric dental residents were in either their first or second year of the residency program. The program is a total of 2 years of graduate training in the care of infants, children, and adolescents after graduating from a 4-year dental program.Infants and young children between the ages of 6 months and 60 months who were enrolled in Head Start programs throughout the five boroughs of New York City were the recipients of oral health assessments and dental care by the interprofessional teams of dental and PNP students.The Head Start programs throughout the five boroughs of New York City were the clinical practice sites for the ICE program for the dental and PNP students. The NYUCD pediatric clinic was also a clinical site for interprofessional oral health assessments, collaborations, and case discussions, among the dental residents, PNP students, and dental and nursing faculty.The Initial Clinical Experience (ICE) course was first developed in 2007, to introduce the dental student to pediatric dentistry and to pediatric caries prevention for children in diverse community settings. It provided dental students with a positive first experience in the role of the health professional. The dental students had the opportunity to clinically examine large numbers of children by performing oral health assessments and preventive treatments, including fluoride varnish to reduce the incidence of dental cavities in young children. The dental students completed a 6-week clinical rotation at various Head Start programs.The ICE program was selected as the ideal program to introduce the PNP and dental students to interprofessional didactic and clinical experience in oral health care for at-risk children. The ICE program was refined to include one PNP student to rotate with eight dental students at various Head Start programs one day a week for a 6-week rotation.Prior to their first clinical practice experience, the students completed the Smiles for Life program  and viewDuring the ICE program, each student was videotaped using an iPad at least two times during the 6-week rotation while performing an oral health assessment and fluoride varnish application. The faculty individually reviewed with each student the videotaped performance on the day of the clinical experience to offer the opportunity to the student to immediately recognize both the strengths of the assessment and areas for improvement.After completing the 6-week rotation in the ICE program, the PNP students took a 10-item written quiz in which the passing grade was set at 80%. If the students did not achieve the passing score or wanted to achieve a higher score, the students had the opportunity to take another quiz on the same topic to show improvement in the knowledge base or to achieve 100% on the quiz.In addition, the PNP students demonstrated their oral health core competencies via an oral health simulation laboratory experience in which each student was presented with a case scenario and then performed an oral health assessment, fluoride varnish, and referrals, as needed, and provided anticipatory guidance to the “Standardized Parent” for the child.Exam questions for the dental students were included in the semester final examination. Since 325 dental students participated in this experience over the course of the semester, their core competencies were evaluated using the iPad technology videotape and then by their respective dental faculty.Students in all health care professions spend much of their time presenting cases to clinical faculty, although the case presentation format and dynamics have been minimally studied , 14. TheSocialization among interprofessional students and faculty is a significant part of establishing the foundation for interprofessional team-based care after graduation. Socialization was fostered by scheduling about 15 minutes before the interprofessional collaborative case presentation for the students and faculty from each profession to meet and socialize in a collegial environment. After the case presentation, students and faculty again had about 15 minutes to socialize.The following data were collected: the number of PNP and dental students who participated in the ICE program over a two-year, two-semester timeframe; the number of children in the Head Start program that received oral health assessments and fluoride varnish applications by the dental and PNP students; oral health assessments videotaped of each student in the ICE program using iPad technology; the number of students who performed collaborative case presentations and the number of students and faculty who attended the presentations; faculty evaluation of all case presentations; the results of the oral health 10-item assessment quiz administered to the PNP students at the end of the 6-week rotation; pass/fail results for PNP students after completion of the oral health simulation laboratory experience; pass/fail results on the oral health assessments and application of fluoride varnish by the dental students in the ICE program.Thirty PNP students, 325 dental students, and 18 residents participated in the program that was completed over a 2-year, 2-semester timeframe. In the first year of the program, 1,642 children in Head Start programs throughout the five boroughs of New York City received oral health assessments and fluoride varnish applications. If a parent was present, the parent received oral health anticipatory guidance. If the parent was not present, a letter was sent home to the parents advising the parents of the child's oral health status and whether further treatment was needed. In the second year of the program, 2,254 children received the same treatments. On average 30 children received oral health assessments and applications of fluoride varnish each day at various Head Start programs. Each student performed approximately five (5) oral health assessments and applications of fluoride varnish each day during their clinical rotation.Each year, two dental residents and three PNP students presented two comprehensive collaborative case studies to dental students in the pediatric rotation, the pediatric residents, and faculty. Lunch was provided to the students and faculty to encourage interprofessional socialization. Fifty dental students, 18 residents, and 15 dental and nursing faculty and deans attended each presentation. Scholarly interactive discussions among students and faculty followed each presentation.All 30 PNP students achieved at least the 80% passing score on the first attempt on the 10-item oral health quiz. Fifteen students received 100% on the first attempt; 10 students received 90%; and 5 received 80%. The 15 students who received less than 100% all took the second 10-item quiz and all but one student achieved a score of 100%.All 30 PNP students participated in the simulation experience. All PNP students were able to demonstrate appropriate oral health assessments, application of fluoride varnish, and appropriate oral health anticipatory guidance based on the presented simulation.All 325 dental students passed the ICE course and demonstrated competencies in oral health assessments and fluoride varnish application.The faculty-student socialization at the end of the 6-week ICE rotation led to a number of student reflections concerning interprofessional clinical practice . All stuCore competencies for PNP and dental students in the area of oral health were identified through faculty analysis of students' performances during the ICE experiences using the iPad videotape. From the documentation of the faculty-student luncheon meetings, another essential interprofessional core competency emerged: oral-systemic health-illness cultural competencies that affect professional attitudes to effectively change population behaviors that promote wellness.Primary care office based assessments are an essential part of the interprofessional initiative to reduce the incidence of early childhood caries (ECC). In addition to the interprofessional oral health performance and affective core competencies (see Tables New interprofessional oral health core competencies for general dental students and pediatric dental residents need to focus on acquisition of knowledge, skills, and attitudes to care for infants and toddlers. New knowledge included oral-systemic health maintenance and anticipatory guidance for parents of infants and toddlers. Establishing a dental home by the infants' first birthday requires that dental students and residents be knowledgeable about infant nutrition and breast feeding guidelines including multivitamin intake for breast feeding mothers and for formula-fed infants. The dental students and residents must be aware of ever-changing trends in feeding infants and young children and how to advise parents about such trends based on scientific evidence. The recent trend by parents to feed infants and toddlers goats milk or raw milk presents challenges to all providers. Dental providers need to keep abreast of these trends and be prepared to discuss the potential oral-systemic consequences of such trends.Managing the anxious child is a critical skill for general dental students who plan to practice as family dental provider. In addition, communication techniques to reassure parents of young children about the significance of oral health care are an essential skill. Pediatric dental office personnel and dentists will need to provide parent education in “one voice” with pediatric primary health care providers so that parents hear one consistent message about oral health care at each dental visit and at each pediatric health maintenance visit.This nonexperimental descriptive study to observe and describe interprofessional educational activities for pediatric dental students, residents, and PNP students has led to the identification of interprofessional performance and affective oral health core competencies for dental and pediatric primary health care providers. Inclusion of these core competencies as part of the curriculum in general dentistry programs and in pediatric and family nurse practitioner programs has the potential to improve the oral health care outcomes for at-risk infants and young children. Therefore, nurse practitioner and dental faculty need to enhance their current curriculum to enable students to be educated in interprofessional classrooms and to practice side by side in health care environments that provide opportunities to attain these oral health core competencies.The ICE program provided the dental and PNP students with a positive first experience in interprofessional education and clinical practice and provided them with an opportunity to clinically perform oral health assessments on large numbers of infants, toddlers, and preschoolers. Caring for diverse populations of children requires health care professionals to be passionate about delivering comprehensive care that is culturally sensitive and focused on ways to educate parents to modify practices that negatively impact the oral-systemic health and well-being of infants and children. Faculty are responsible for cultivating reciprocal professional referrals among dentists who perform restorative procedures and pediatric primary care providers who not only refer children and parents for the procedures but also support parents by informing them of the need to improve oral-systemic health in their children. Behavior management for toddlers, preschoolers, and school-age children is another essential core competency that needs to be acquired by dental students, pediatric dental residents, and pediatric nurse practitioners. Gaining the cooperation of the child during primary care and dental office visits reduces the fears displayed by children in unfamiliar environments.The use of the iPad videotapes proved to be valuable to each individual student. The student-faculty review showed the student immediate corrections that were needed to improve the performance of the oral health skills. The student then had the opportunity at the next clinical experience to perform the skill again and view the corrections.The interprofessional socialization and discussion at the end of the 6-week ICE program and the case presentations provided unique opportunities for the dental and PNP students to collaboratively manage both simple and complex oral health care issues. Consistent with the findings of Thistlethwaite et al. , the stuIt has been 13 years since the compelling report Oral Health in America by the Surgeon General which identified the poor oral health status of children from lower socioeconomic and diverse populations . Impleme


Coffea arabica plants growing in the rustic plantations and simple polycultures of two regions in the center of Veracruz, Mexico. The endophytic fungi isolates were identified by their morphological traits, and the majority of identified species correspond to species of fungi previously reported as endophytes of coffee leaves. We analyzed and compared the colonization rates, diversity, and communities of endophytes found in the different agroforestry systems and in the different regions. Although the endophytic diversity was not fully recovered, we found differences in the abundance and diversity of endophytes among the coffee regions and differences in richness between the two different agroforestry systems of each region. No consistent pattern of community similarity was found between the coffee agroforestry systems, but we found that rustic plantations shared the highest number of morphospecies. The results suggest that endophyte abundance, richness, diversity, and communities may be influenced predominantly by coffee region, and to a lesser extent, by the agroforestry system. Our results contribute to the knowledge of the relationships between agroforestry systems and biodiversity conservation and provide information regarding some endophytic fungi and their communities as potential management tools against coffee plant pests and pathogens.Over the past 20 years, the biodiversity associated with shaded coffee plantations and the role of diverse agroforestry types in biodiversity conservation and environmental services have been topics of debate. Endophytic fungi, which are microorganisms that inhabit plant tissues in an asymptomatic manner, form a part of the biodiversity associated with coffee plants. Studies on the endophytic fungi communities of cultivable host plants have shown variability among farming regions; however, the variability in fungal endophytic communities of coffee plants among different coffee agroforestry systems is still poorly understood. As such, we analyzed the diversity and communities of foliar endophytic fungi inhabiting  Coffea arabica L. Commercial coffee production relies mainly on the plant species The relationship between coffee agroforestry types and their biodiversity has been attracting attention over the past two decades Studies on the fungal diversity associated with Mexican coffee plantations, especially arbuscular mycorrhizal fungi (AMF) Other studies on the foliar endophytic fungi (EF) associated with coffee plants have been conducted in abandoned coffee plantations in Puerto Rico Coffea arabica, the fungal endophytes are transmitted horizontally Endophytic fungi are microorganisms that live in plant tissues without causing apparent harm to the host The EF communities (EFC) may be influenced by the traits of a given host plant Coffea arabica leaves. We analyzed and compared the colonization rates (CR), richness, diversity, and fungal communities of the EF inhabiting the leaves of Coffea arabica in different agroforestry systems in two different coffee regions, Huatusco and Coatepec, in the center of the state of Veracruz, Mexico. We selected coffee plants from a rustic plantation and from a simple polyculture in each of the coffee regions. We isolated 471 foliar endophytes, which we assigned to 31 morphospecies, from the four selected coffee plantations. We found differences in CR and EF diversity between the coffee regions, as well as differences in richness between the two different agroforestry systems of each region. The analysis of EFC similarity revealed that the endophytic communities in the two coffee plantations in Coatepec did not vary, while the EFC of the coffee plantations in Huatusco were different. There were similarities in the EFC of the rustic plantations and in the EFC of the simple plantations.The aim of the present study was to analyze whether the coffee agroforestry system and the region where the coffee is cultivated could influence EF diversity and the EFC associated with Studying the fungi associated with the phyllosphere of coffee plants enable us to recognize the presence of coffee pathogens and to evaluate, in future studies, the potentiality of some EF and EFC in controlling pathogens or pests of this important cultivated plant.No specific permits were required for the described field studies. This work did not involve endangered or protected species.Four shaded coffee plantations from the central coffee-growing region of Veracruz, Mexico were selected as field study sites . Two plaHeliocarpus sp., Quercus sartorii, Myrsine coriacea, and Trema micrantha, with some selected shade trees, such as Inga spp. and Citrus spp. The canopy has a high degree of vertical stratification, and the vegetation contains abundant epiphytes In rustic plantations, coffee is grown under the shade of native vegetation, such as Inga spp.), and crop species with commercial value, such as Citrus spp. and Musa sp. In simple polyculture coffee plantations, the canopy of the native trees of the forest are removed and replaced with selected shade trees, such as legumes  plates. The EF that did not sporulate on these media were transferred to PDA, OA, and malt extract agar (MEA 2%) plates and to plates with leaf extracts of healthy, mature coffee leaves (10% wt/vol) to activate sporulation.We examined the above-mentioned traits of the mycelium morphology of the fungi isolates and their microscopic characteristics. The characteristics evaluated for the anamorph type were conidiomata, conidiogenous cells, conidiophores, and conidia morphology , and the characteristics evaluated for the teleomorph type were sporomata, their associated structures, and spore morphology. The strains of isolated and identified endophytic fungi are part of a fungal collection of the Laboratorio de Alelopatía, Departamento de Ecología Funcional, Instituto de Ecología, Universidad Nacional Autónoma de México; the strains are freely available.Colletotrichum genus were cultured in basal media supplemented with a different carbon source . Using that media enabled us to evaluate the substrate utilization of the isolated Colletotrichum strains and differentiate among species, as some species, such as the coffee pathogen Colletotrichum kahawae, cannot metabolize tartrate and citric acid Following the method described in Waller et al. (1993) The fungi were classified into morphospecies based on their growth rates and morphological and microscopic characteristics. Isolates that did not sporulate were identified as morphotypes (Mycelia sterilia), based on their morphological characteristics. The relative frequency of EF for each coffee plantation was calculated as the abundance of a given species divided by the total number of fungi isolated from each coffee plantation.Colonization rate (CR) was calculated as the number of fragments from which one or more EF was isolated, divided by the total number of incubated fragments CR and IR were analyzed using a two-way ANOVA test, with regions (Huatusco and Coatepec) and agroforestry category (rustic and simple polycultures) as the factors. The difference between mean values was evaluated using Tukey's honestly significant differences (HSD) test. The statistical analysis was performed using Statistica software 8.0.Because leaves serve as largely discrete, relatively uniform, bounded habitats to the microbes that inhabit them The richness and distribution of EF found in each coffee plantation were examined using the range diversity (RD) analysis developed by Arita et al. (2012) Following the R script of RD analysis sp) and for leaves (Vlv) according to the method of Arita et al. (2012) sp were contrasted with null models in which we maintained the original frequency of EF richness found in every coffee plantation, but we assigned leaves to species randomly. The empirical values of Vlv were contrasted with null models in where we maintained the original frequency of number of species found in each coffee leaf, but we generate permutations to simulate the random assignment of species to leaves.To test a possible association among species and a possible clustering of leaves in terms of shared species in each coffee plantation, we computed the ratio of variance for species  indexes of fungal endophyte species found in each coffee leaf were calculated with EstimateS software EFC similarities found among the coffee plants from the four plantations were analyzed using non-metric multidimensional scaling (NMDS) plots. The NMDS plots display the dissimilarities among EF communities graphically, and the distances between them on the plot represent their relative dissimilarity Coffea arabica leaves collected. At least one EF was isolated in each the leaves examined, with one exception: no fungi were isolated from one coffee leaf from the simple polyculture plantation in Coatepec (SP-C). Therefore, that leaf was not included in the statistical analysis. The ANOVA test showed significant differences in CR and IR between the two regions  and between the two agroforestry types . According to the post-hoc tests  based on spore and conidiogenous cell size and mycelium traits. In the Xylaria genus, we recognized six morphotypes according to their morphological characteristics. Due to a lack of spore production, eight EF morphotypes, which represented only 5% of the isolated fungi, could not be identified; they were named Mycelia sterilia 1–8.The Xylaria was isolated withered after a few days of culture, while the leaf segments colonized by Coniosporium remained green in the Petri plates for almost one month.Interestingly, we observed changes in the appearance of the fresh leaf fragments cultured in the Petri plates according to the fungi isolated from them. The leaf segments in which the genus Accumulation curves of observed species richness (gray triangles and circles) and Jackknife 1 estimated richness (open triangles and circles) are shown in The only coffee plantation in which the rarefaction curve reached the saturation point was the SP-C . RichnesThe rarefaction curves of the other three plantations did not reach the saturation point, suggesting that endophyte richness was not fully recovered. The Jackknife 1 expected richness for those locations was higher than the observed richness, indicating that species richness was not exhaustively sampled.The results of the RD analysis summarized in Colletotrichum gloeosporioides 1.On average, each EF morphospecies was isolated in five leaves from the Huatusco plantations and in four leaves from the Coatepec plantations. The minimum number of leaves in which a given morphospecies was isolated was one leaf, and the maximum was 14–20 leaves. The most widespread EF species in all four coffee plantations was sp value, which indicates the degree of association among different species sp of null models with 10 iterations . Empirical and null models showed no association among morphospecies in those plantations. In RP-C and SP-H, the empirical Vsp values were less than 1, while in the simulations values Vsp were 1 . Those results indicate negative associations among EF species in RP-C and in SP-H. Negative covariances were observed between Colletotrichum gloeosporioides and Xylaria, Xylaria and Coniosporium sp., and Colletotrichum gloeosporioides and Coniosporium sp. In general, positive covariances were found among fungi isolated in low frequencies, such as Paecilomyces sp., which was only isolated from leaves in the presence of one Coelomycete endophyte. The Vlv value, which indicates EFC similarity among leaves from the same coffee plantation, was higher than 1 in all four coffee plantations , and indicates considerable similarity in the shared species of the leaves of each coffee plantation. The highest EFC similarity was found in the coffee leaves from SP-H.The Vntations . Those rThe ANOVA test of diversity indexes of EF isolated from the coffee leaves showed significant differences in diversity between agroforestry systems  and with the interaction between coffee region and agroforestry system . The Shannon's diversity was different between coffee regions , while the Fisher's alpha was not different between them  due to the variability in Fisher's alpha diversity in Huatusco region. Fisher's alpha EF diversity  was signThe NMDS plots  with JacThe results of the similarity analysis summarized in The ANOSIM analyzed with the Euclidean distances also showed significant differences between the RP-H EFC and the SP-C EFC (R = 0.186). The NMDS plot based on Euclidean distance shows that the circles representing EFC from SP-C  and in the EFC of the simple plantations (SP-H and SP-C).In this study, we identified 31 morphotaxa from the 479 isolated endophytic fungi. The number of morphospecies found in our study is lower than the number found in the study by Santamaría and Bayman (2005) Colletotrichum and Xylaria, which have often been isolated from coffee plants in American coffee-growing regions PhomopsisBeauveriaAlternariaConiosporiumGuignardiaPaecilomycesDiplodia, BotryosphaeriaThe species reported in the present study are ubiquitous taxa isolated mainly from plants inhabiting tropical regions Colletotrichum gloeosporioides complex includes strongly aggressive pathogens, opportunistic pathogens Colletotrichum gloeosporioides is associated with blister spot, or “mancha mantecosa,” in leaves Colletotrichum gloeosporioides in this study, as well as the successful inoculation of these fungi in coffee plants without causing evident damage or disease , show the endophytic nature of this species isolated from coffee leaves.Knowledge of the diversity of EF in coffee leaves is very important in identifying phytopathogenic fungi that might live as endophytes during part of their life cycle Xylaria is recognized as a saprotrophic fungus Xylaria was isolated withered in a few days of Petri dish cultures, and some of them produced stromata only over the dead leaf segment. This phenomenon might be evidence of the saprobic phase of the xylariaceous fungus The genus Beauveria, which was only isolated from RP-H leaves, and Paecilomyces, are recognized as entomopathogenic fungi and have been isolated and tested on some coffee pests Some genera, such as The analysis of EF abundance in the coffee plantations in the two regions studied indicated higher CR and IR values in the Huatusco plantations than in the Coatepec plantations. This finding indicates that the geographical region  exerts an important influence on the colonization and abundance of foliar EF in coffee plants. Other studies have also shown differences in the abundance of EF between different localities in agrosystems On the other hand, we found no significant differences in CR or IR between the two agroforestry systems of each coffee region; however, we found higher CR and IR values in the rustic plantations than in the simple polycultures. Coffee plants in rustic plantations, as opposed to simple polycultures, are grown under a higher diversity of shade trees species, representing a more complex canopy. However, as shown in other studies, the canopy does not seem to influence the colonization and abundance of EF in plants growing in the understory Vitis vinifera) cultivated in vineyards following different farming practices.Besides the differences in canopy, the two coffee agroforestry systems also differ in their farming practices. However, the differences in management systems also do not seem to affect the colonization and abundance of EF. Similarly, Pancher et al. (2012) We found that the CR and IR of EF are not necessarily related to the diversity found in each sampling site. For example, Matsumura and Fukuda (2013) We found a higher total EF richness, a higher mean richness by leaf, and a higher expected richness in the rustic plantation than in the simple polyculture of each region. As mentioned previously, these results suggest that the complexity of the canopy and the alternative management techniques used in rustic plantations might influence the EF richness and diversity. Some studies have shown that the canopy might influence the richness and diversity of foliar EF growing in the understory Various studies have been conducted on biodiversity conservation in different coffee production systems in the central region of Veracruz, where Huatusco and Coatepec are located In addition to the variability in richness and diversity of EF between coffee agrosystems, we found higher EF richness and diversity in RP-H than in the other three plantations, and a higher mean richness by leaf in the Huatusco plantations than in the Coatepec plantations. One reason for this finding is the difference in EF distribution in the two regions. Although we observed the typical distribution of EF reported in many studies on tropical and temperate forest trees The Fisher's alpha and Shannon's index diversity analysis results differed in the present study. Both indexes consider the number of EF species (richness) and relative abundance of the individuals present in a given sample, but Fisher's alpha is not influenced by the sample size and is less affected by the abundance of the most common species than Shannon's index Fagus sylvatica) Although the total EF diversity found in the coffee plantations was not fully recovered, we found similar diversity values in other studies about EF diversity. In the present study, the mean of Fisher's alpha diversity of morphospecies (5.1–6.5) is similar to the mean of genotypic diversity  of coffee plants in Mexico The Shannon's diversity values varied around 2 in the four coffee plantations. According to Gazis and Chaverri (2010) Colletotrichum gloeosporioides 1 and Xylaria 1, have more negative co-occurrences than species isolated in low frequencies. The negative covariances between Colletotrichum gloeosporioides and Xylaria spp. have been reported previously for endophytes inhabiting coffee leaves In the analysis of association between species, the interactions between fungi are apparently different in each coffee plantation, but in general, the fungi isolated in high frequencies, such as Species found in high frequencies co-occur with a low number of species, whereas the species isolated in low frequencies co-occur with a high number of species. Our results are consistent with those of Pan and May (2009) The NMDS plots based on the different ecological indexes showed variation in EFC among the different plants of each coffee plantation. In addition, there was no EFC clustering among coffee plantations. When the EFC similarities of the coffee plantations were compared with an ANOSIM test, we found that the geographically closest plantations, the ones in the Coatepec region, were similar in their EFC, independent of their agroforestry systems. In agreement with this result, some studies have shown that EFC similarity is a function of the distance between sites, and thus, nearby sites have similar EFC In contrast, we found dissimilarities in the EFC of the two Huatusco plantations, even though they were near each other. The ANOSIM test showed that the EFC of RP-H and SP-H were different according to the three ecological indexes of similarity that were analyzed. The EFC of RP-H and SP-C were also different as analyzed using Euclidean distance. Those results suggest that agroforestry system could influence the assemblage of EFC of coffee plants. Our results also showed that the EFC of the rustic plantations and the EFC of the simple polycultures were similar; in fact, the rustic plantations shared a higher number of morphospecies than the simple polycultures did.Vitis vinifera plants under different vineyard management practices. These results indicate that the plantation management techniques in agroforestry systems, such as coffee plantations and vineyards, might influence the assemblage of EFC.Studies on the EF of herbaceous cultivable plants, such as cotton and maize, have shown no similarities in EFC based on farming practice More sampling will help elucidate the factors that influence the EFC, but as shown in this study, some of those factors might be the geographical location and agroforestry system of the coffee plantations. Further studies evaluating the ecological roles of the different EFC found in the diverse coffee agroforestry systems will contribute to the knowledge of the role of the system of agroforestry management in the regulation of pest and pathogen populations The results of the present study show that the coffee agroforestry system produces variability in the colonization, richness, diversity, and composition of EFC in coffee plants. In addition, they demonstrate that the region in which coffee is cultivated is an important factor that influences these parameters. Future studies on the biodiversity conservation value of different coffee agroforestry systems will need to consider the coffee region as a determinant factor that affects biodiversity. In further studies, the use of molecular and physiological tools to identify, individually and collectively, the functional and ecological significance of EF in coffee plants under diverse ecological and geographical conditions will be equally significant. These studies will also provide an opportunity to understand the potential use of some EF as producers of relevant precursor substances in the regulation of different pests and pathogens, to discover new drugs, and to understand the potential role of EFC as potential controls of pest populations.Table S1Description of morphospecies of foliar endophytic fungi isolated from the four coffee plantations.(DOCX)Click here for additional data file.


Despite the dynamic nature of ecological interactions, most studies on species networks offer static representations of their structure, constraining our understanding of the ecological mechanisms involved in their spatio-temporal stability. This is the first study to evaluate plant-herbivore interaction networks on a small spatio-temporal scale. Specifically, we simultaneously assessed the effect of host plant availability, habitat complexity and seasonality on the structure of plant-herbivore networks in a coastal tropical ecosystem. Our results revealed that changes in the host plant community resulting from seasonality and habitat structure are reflected not only in the herbivore community, but also in the emergent properties (network parameters) of the plant-herbivore interaction network such as connectance, selectiveness and modularity. Habitat conditions and periods that are most stressful favored the presence of less selective and susceptible herbivore species, resulting in increased connectance within networks. In contrast, the high degree of selectivennes  and modularity of the networks under less stressful conditions was promoted by the diversification in resource use by herbivores. By analyzing networks at a small spatio-temporal scale we identified the ecological factors structuring this network such as habitat complexity and seasonality. Our research offers new evidence on the role of abiotic and biotic factors in the variation of the properties of species interaction networks. Approximately 40% of terrestrial biodiversity is involved in plant-herbivore species interaction networks Despite the advances in the identification and description of structural patterns of ecological networks, relatively little is known about the ecological and phylogenetic mechanisms that give rise to such patterns Furthermore, both seasonal changes in precipitation and structural complexity of plant communities are intimately related with changes in microclimate (e.g. temperature and humidity) and with the availability and quality of host plants for herbivores Addressing the spatio-temporal assessment of ecological networks represents a new frontier in community ecology This study is unique in the sense that, for the first time, the impacts of resource availability, habitat structural complexity and seasonality on the structure (parameters) of an antagonistic network were assessed.Field work was carried out at La Mancha Coastal Research Center  located on the coast of the state of Veracruz in Mexico . Permit to perform the present research was issued by Instituto de Ecología, A.C. (INECOL), since CICOLMA is one of INECOL's field station and private conservation reserve. CICOLMA is a very small fraction of the 1336 Ramsar Site called “La Mancha el Llano” declared in 2005. Thus, the regulatory body concerned with protection of wildlife is given to INECOL by Mexican federal organisms in the matter and framed by The Ramsar Convention on Wetlands . The study site, the CICOLMA reserve area managed by INECOL covers an area of 70 ha, which includes several native vegetation types (including wetlands and dune vegetation), transformed forestry/agricultural sites, experimental areas and a field station. The major vegetation types here are tropical deciduous forest, tropical dry forest, sand dune scrub, mangrove forest, freshwater marsh and flooded deciduous forest The climate of the study site is warm sub-humid and total annual precipitation ranges from 899 to 1829 mm, ca. 78% of the total annual precipitation falls during the rainy season (June - September). In this area, 837 plant species have been recorded, 50% of which are herbaceous plants and the rest are shrubs, trees and vines The study was carried out at five sites with vegetation types representative of the plant communities in the study region and with differing degrees of structural complexity: 1) pioneer dune vegetation (PIO), 2) coastal dune scrub (DUN), 3) recently established tropical lowland subdeciduous forest (SFY) 4) tropical lowland subdeciduous forest in an advanced stage of succession (SFO), 5) tropical lowland flood forest with a wetland ecotone (FFW).Structural complexity and vegetation strata differ specially between the open  and the forest habitats . The open habitats have moderate vegetation cover due to increased sand movement at the dunes, with dominant herbaceous stratum .These habitats are exposed to abiotic stressful conditions because of high temperatures, radiation, salt water influence and mechanical damage of vegetation during storms and hurricanes At each of these vegetation types we placed 14 quadrats (4×4 m) where plant-herbivore interactions were recorded, along with the attributes of each plant community. In monthly censuses during the dry  and the wet/rainy  season of 2012, we collected all folivorous caterpillar species (excluding leaf miners) associated with host herbs, lianas, shrubs and trees. In order to search for caterpillars on adult trees, a subsample of four branches was taken per individual tree. All caterpillars were recorded and taken to the laboratory where they were fed until pupated. No ethical approval was required for manipulating lepidopteran specimens, since after caterpillars pupated and adults emerged, only unidentified species  were mounted as adults for taxonomical identification . Data underlying the findings of the present study are available within this manuscript, but lepidopteran specimens found at the present study are being processed for subsequent molecular analysis  after which specimen identity will be publicly available in reference to this manuscript. Host plants were identified and deposited in the XAL Herbarium of the Instituto de Ecología, A.C. .To assess spatio-temporal changes in the structural network parameters, for each of the five sampled vegetation types we constructed four matrices of the plant-caterpillar interactions throughout the dry and wet season: dry season 1 , dry season 2 , rainy season 1  rainy season 2 . Previous work at the study site suggests that there is short-scale temporal variation in the structure of insect-plant –mutualistic- interaction networks derived from changes in abiotic properties (temperature and precipitation) 2), given that H2 describes to which extent observed interactions deviate from those that would be expected given the species marginal sums 2) is based on the deviation between the observed and expected number of interactions in each network, assuming that all species interact with other species in proportion to their total observed frequency. Given that this metric is not affected by neither sampling effort nor network size, it is possible to make robust, reliable comparisons between different networks 2 values, ranging from 0 (low selectiveness) to 1 (high selectiveness). The network indices were calculated for each census and site using the BIPARTITE 3.0.1 package run in R We obtained a total of 20 matrices of species interactions , which we refer to as sub-networks. We calculated the following structural parameters for each sub-network: connectance (C), Interaction evenness (IE), vulnerability (V), generality (G), modularity (M), number of modules (NM) and network specialization or selectiveness (selective use of the lower trophic level by the higher trophic level) of the entire bipartite network  was registered, along with an estimation of canopy cover provided by all plant species (included host and non-host species) found in each quadrat . As an indicator of food availability to herbivores, we measured plant cover of host species . Species richness of host plants used by herbivorous lepidopterans (host plant richness  = HPR) was also registered. These variables were selected because they influence herbivore food availability, habitat structure and the abiotic factors that affect plant-herbivore interactions.2) were evaluated using repeated measures ANOVAs, using the sub-network parameters as replicates for each vegetation type and census. Using a standard ANOVA in this case is not appropriate because it fails to model the correlation between the repeated measures (census and vegetation types): the data violate the ANOVA assumption of independence. Therefore the repeated measures ANOVA is suitable for analysis of auto correlated samples. To assess the influence of vegetation type, the model included census as the error term (within subject) and vegetation type as the explanatory variable (between subject). To evaluate the influence of census, the model included vegetation type as the error term (within subject) and census as the explanatory variable.The effect of vegetation type and census on plant community attributes  and structural parameters of each network . Structural network parameters were subsequently estimated for each sub-network. In order to describe the dissimilarity among these matrices, we performed a NMDS analysis . The resulting NMDS ordination (biplot) shows the structural parameters of all sub-networks: when sub-networks are close to a specific parameter, these have small dissimilarities in their values for that parameter. This is one of the most effective methods for the ordination of ecological data and the identification of underlying gradients, because it does not assume a linear relationship among the variables To evaluate if spatio-temporal dissimilarity among sub-network parameters was related to plant community attributes, we analyzed the effect of TPR, TPC, HPC (continuous variables) (see “Attributes of the plant community” within this section) on the ordination of plant-herbivore sub-networks using the envfit function from the VEGAN library of the statistical package R adonis function of the VEGAN package run in R. This function runs a multivariate analysis using the distance between matrices (dissimilarities between objects or sampling sites), and the probability of significance P is obtained through permutations To test the null hypothesis of a similar composition of herbivore and host plants species among vegetation types and sampling censuses, we used a PERMANOVA , and the Bray-Curtis Index 2, M y NM (quantitative variables or vectors) over the ordination (biplot) of an NMDS analysis of the community of lepidopteran herbivorous larvae found foraging on plants at the 20 sub-networks using the envfit function from the VEGAN package of the statistical software R. This function provides a correlation measure and is statistical significance .To understand if spatio-temporal changes in herbivore species composition (ordination dissimilarity) could affect network structure, we analyzed the effect of parameters C, H2 of the network varied significantly across censuses and among vegetation types . For both herbivores (lepidopteran larvae) and hosts plants, species turnover was higher between groups (vegetation types) than within them . The mean values of C, M, NM and Hon types . The neton types .2 of the forest habitats  were significantly higher than those with open vegetation   , Table 2IO, DUN) .2 values that were twice as much of those estimated for the networks of the first census. None of these indices were different among the last three censuses  and vector HPC . The correlation of HPC was greater for the ordination of the sub-networks in habitats with open vegetation and during the dry season . TPR hadThe NMDS exploring the relation among sub-network parameters and herbivore species composition showed an important spatio-temporal turnover of lepidopteran herbivore species . IndepenIt has recently been demonstrated that the structure of plant-herbivore interaction networks is influenced by ecological, phylogenetic Our results show for the first time how the structural parameters of plant-herbivore networks are spatially and temporally dynamic, influenced by seasonality, habitat complexity and food resource availability. The spatio-temporal changes in host plant communities resulting from abiotic factors  were not only reflected in the species composition of herbivore community as previously reported In contrast with mutualistic plant-insect networks, which maintain their structural stability over time 2) metric is not affected by neither sampling effort nor network size 2 pattern among sub-networks suggest that, independently of the number of species constituting each sub-network, there is higher selectiveness of lepidopteran larvae  during the rainy season when offer of plant species (and foliage availability) is higher; this pattern may reflect changes in host choice by adult females, as a function of temporal changes in plant community Although connectance is a simple parameter sensitive to network size The ordination of the most specialized and highly modular sub-networks was highly correlated with floristic richness of forest habitats and the rainy season. The high degree of specialization and modularity of the networks in the richest floristic contexts could be related to a greater diversification by the herbivores in their use of plant resources: a greater availability of niches at the lower trophic level is likely to promote a greater niche differentiation  in the higher trophic level. The forest sites  were not only floristically the richest of the studied sites, but also offered more canopy cover from trees and shrubs, as well as more plants with different life forms The highest sub-network connectance was found under the most stressful abiotic conditions: Open vegetation and the dry season create conditions that could cause greater water stress in plants as a result of high temperatures and solar radiation. In turn, this could affect their nutrition (water and nitrogen content) and a defense (toughness and phenolic compounds content) attributes rendering the plants unsuitable for many lepidopteran herbivores Chamaecrista chamaecristoides, Crotalaria incana and Porophyllum punctatumAt the present study most generalist herbivores were associated with host plants with large plant cover, mainly pioneer species adapted to stressful natural habitat conditions such as In antagonistic networks, connectance - whether promoted by the range of host plants eaten by herbivores or by resource availability- can mitigate the loss of species when environmental conditions are unstable or stressful In the studied plant-herbivore sub-networks, higher connectance was detected during the dry season in the pioneer habitat, and this seems similar to the changes that other trophic networks undergo when subjected to anthropic disturbances. The way in which the loss of arboreal cover reduced network complexity has been reported as a process of spatio-temporal homogenization of species and their interactions An important property of our plant-herbivore network was the spatio-temporal prevalence of vulnerability and interaction evenness. Regardless of habitat type or season, the ratio of herbivore to plant species was the lowest and no dominant interactions were recorded. Our results are similar to those of other trophic networks for habitats with differing degrees of fragmentation and where there was no evidence of variation in the structural attributes of the network In recent years, there have been efforts to assess the spatio-temporal variation of ecological networks


We recently reported an increased risk of serious pneumococcal disease (SPD) in asthmatics. Little is known about the impact of asthma status on the severity of SPD. We compared the severity of serious pneumococcal disease (SPD) between patients with asthma and those without asthma. The study subjects were Rochester, Minnesota residents who developed SPD between 1964 and 1983. SPD and asthma status were ascertained by using explicit predetermined criteria Severity of SPD was assessed using intensive care unit (ICU) admission rate and total days of ICU stay and hospitalization associated with treatment for SPD. We found that there were no significant differences in severity outcomes between asthmatics (n=11) and non-asthmatics (n=163). Asthma status may increase the risk of SPD but not influence its severity. However, given a small sample size of our study, a larger study needs to be considered to clarify the relationship between asthma and severity of SPD. A previous study assessed underlying conditions among patients aged 2–64 years who developed invasive pneumococcal diseases n=3,469) before the introduction of hepavalent Pneumococcal Conjugate Vaccine (PCV-7) . Only 50,469 befoGiven the increased risk of development of Serious Pneumococcal Disease (SPD) in individuals with asthma, whether asthma status influences severity of SPD may be an important question to be addressed because severity and risk of infection can have different risk factors and mechanisms. Addressing this question may be important in determining the prognosis and management of SPD when asthmatics acquire SPD. Currently, there is no population-based study that examined the relationship between asthma and the severity of SPD. To determine the association between asthma status and severity of SPD, we conducted a retrospective cohort study. In this study, we assessed the impact of asthma status on the severity of SPD in the same study population during the primarily pre-pneumococcal vaccine era, 1964–1983 who was studied for the association between asthma and the risk of SPD.The study was approved by both the Institutional Review Boards at Mayo Clinic and Olmsted Medical Center. This is a population-based retrospective cohort study designed to assess whether asthma status is associated with the severity of SPD among the Rochester residents who developed SPD between 1964 and 1983, a primarily pre-pneumococcal vaccine era. SPD cases were identified through reviewing 3,941 medical records and asthma status was ascertained by predetermined criteria for asthma .The study setting and population were previously described in detail . BrieflyStreptococcus pneumoniae from a normally sterile site such as blood or cerebrospinal fluid), and/or pneumococcal pneumonia requiring all three of the following criteria, 1) a physician diagnosis of pneumonia, 2) the isolation of pneumococcus from sputum gram-stain or culture, and 3) the documented pneumonia by chest radiograph. We defined the index date of onset of the SPD as the date of documented isolation of S. pneumoniae.We reported the details for identification of SPD cases previously . BrieflyAfter all SPD cases were identified and confirmed, we used the previously collected data on asthma during the period 1964 to 1983 to ascertain the asthma status among the confirmed SPD cases  7]. In . In 7]. We used the number of days of inpatient hospitalization due to SPD as the primary surrogate measure for severity of SPD. A previous study showed that the number of days of inpatient hospitalization was well correlated with severity of IPD . We defiSocio-demographic and clinical characteristics of SPD cases with and without asthma were descriptively summarized . The totA total of 3,941 records were reviewed and we identified 174 SPD cases. Of the 174 SPD cases, 11 cases (6.3%) were asthmatics. Socio-demographic and clinical characteristics of SPD cases with and without asthma are summarized in There were no significant differences in SPD severity between asthmatics and non-asthmatics . The perOur previous study showed an increased risk of SPD in asthmatics. However, the current study results suggest that there were no significant differences in all outcome measures for severity of SPD between asthmatics and non-asthmatics. The literature on IPD suggests that persons with more severe illness, such as meningitis, had longer duration of hospitalization compared with those who had less severe illness such as bacteremia or pneumonia . LaurichWhy does asthma status have a different impact on the incidence and severity of SPD? We can only postulate it. The literature suggests asthmatics have Th2-predominant biological milieu  and haveThe strengths of the study include population-based study design during the primarily pre-pneumococcal vaccine era, self-contained health care system with unified medical record system for research, and independent ascertainment of SPD cases and asthma status. Also, there are inherent limitations in the study due to its retrospective design. We did not measure any laboratory or clinical parameters for severity of SPD. Although the hospital duration is a commonly used indicator for severity, future study needs to include laboratory or clinical measures for severity of SPD. Our study had a small sample size so that the study findings may be subject to type-2 error. Our study subjects were predominantly Caucasian population and it is cautious to generalize our study findings to other study settings.In conclusion, although asthma status has been reported to be associated with the increased risk of SPD, it does not influence the severity of SPD. Given a small sample size of our study, a study with a larger sample size needs to be considered to clarify the relationship between asthma and severity of SPD.


Urea is generated in humans following the breakdown of amino acids and is evenly distributed throughout the body, including in the central nervous system, subcutaneous adipose tissue, blood serum, and epithelial lining fluid 4N2O+H2O→NH3+CH3NO2CH3NO2+H2O→NH3+H2CO3CHUnder physiological conditions the proton of carbonic acid dissociates, and the ammonia molecules become protonated to form ammonium, causing an increase in local pH that can interfere with host function. The role of urease in the virulence of some bacterial pathogens is well established; however, more recent studies are beginning to highlight the function of urease during human fungal infections, suggesting that this enzyme has a wide role during microbial infection.Helicobacter pylori to colonise the acidic environment of the human stomach. In response to the acidification of the periplasm of H. pylori, a proton gated urea channel (UreI) imports urea into the bacterial cytosol, where it is hydrolysed by urease H. pylori is unable to establish infection in the stomach H. pylori infection Urease activity is the basis of acid acclimation that enables Proteus and Klebsiella species. Infection by these urease-positive bacteria can result in the development of infection stones that surround and protect the pathogen. Stones form due to the precipitation of the minerals struvite and carbonate apatite, which are produced by the binding of ammonium to magnesium ions and bicarbonate to calcium ions, respectively H. pylori infection in the stomach, ammonia has an additional role in urinary tract infections by causing damage to the glycosaminoglycan surface of the urothelium that protects epithelial cells from bacterial infection A distinct urease-dependent process is associated with bacterial urinary tract infections, including those caused by Cryptococcus neoformans (a basidiomycete) and Coccidioides posadasii (an ascomycete), suggest that urease has conserved roles in promoting bacterial and fungal infections. The inhalation and subsequent germination of fungal spores is the route of entry into the body for many fungal pathogens. Successful infection of the lung requires the pathogen to evade the host immune system, and subsequent dissemination is dependent on the ability of the fungus to move from the lungs to other organs via the bloodstream. C. neoformans and Co. posadasii are saprophytic yeast that infect humans via the lungs and cause disease, with the severity of infection correlating with a loss of host immune function, and in both cases, the disseminated form of infection is potentially life threatening C. neoformans and Co. posadasii suggest that urease-dependent pH changes are involved in immune system evasion and that ammonia toxicity to host cells promotes systemic disease.Studies of two evolutionarily diverse fungi, C. neoformans and Co. posadasii in animal models of disease C. neoformans have higher levels of serum IgE, Th2 cytokines, and alternatively activated macrophages as compared to control mice infected with a urease-negative strain Co. posadasii infection support this positive feedback model Urease is required for the full virulence of Co. posadasii infection C. neoformansMycobacterium bovis, which localise more efficiently with lysosomes than a parental wild-type strain M. bovis-derived ammonia accumulates in macrophages and correlates with the reduction in the cell surface trafficking of the major histocompatibility complex class II One mechanism to account for the urease-dependent Th2 polarisation of the host immune system is that the pH changes associated with urease activity cause a reduction in the acidification and maturation of phagolysosomes in phagocytic cells, resulting in a loss of pathogen killing and antigen presentation. A range of studies are consistent with this model. Fungi excrete ammonia as part of their internal pH control, and localised increases in pH are associated with C. neoformans. Urease activity negatively influences the integrity of endothelial junctions in vitro, as assayed using human brain microvascular endothelial cells C. neoformans across the blood-brain barrier in vivo after the pathogen becomes trapped in the small capillaries of the mouse brain C. neoformans to migrate into the brain parenchyma. A prediction is that pathogen-derived ammonia has the potential to kill host cells throughout the body.The dissemination of a fungal pathogen from the lungs to other organs will be facilitated by damage to epithelial cells that allow pathogen access to and from the bloodstream. Ammonia is toxic to mammalian cells, and the urease-dependent damage to human epithelial cells by bacterial pathogens suggests that the ammonia produced by urease-positive fungi may also promote fungal dispersal The studies relating to the ammonia repression of phagocyte function and localised tissue damage suggest a general model for the role of urease during fungal infections . InitialH. pylori, Sporosarcina pasteurii, and Klebsiella aerogenes, have culminated in the following model of urease activation Urease was the first protein to be identified as a nickel enzyme C. neoformans and are essential for full urease activity The bacterial mechanism of urease maturation is predicted to occur in fungal cells. Homologues of the bacterial UreDFG proteins (Ure467) and a nickel importer (Nic1) exist in Aspergillus fumigatus, Histoplasma capsulatum, Blastomyces dermatitidis, and Paracoccidioides brasiliensisThe model of urease function during fungal infections is based on studies of two pathogenic fungi and the extrapolation of work with bacterial pathogens. However, nothing is known about the role of this enzyme during infection by other urease-positive fungi that infect the lungs, such as