Use of Pyrosequencing and Denaturing Gradient Gel Electrophoresis to Examine the Effects of Probiotics and Essential Oil Blends on Digestive Microflora in Broilers Under Mixed Eimeria Infection

A protective digestive microflora helps prevent and reduce broiler infection and colonization by enteropathogens. In the current experiment, broilers fed diets supplemented with probiotics and essential oil (EO) blends were infected with a standard mixed Eimeria spp. to determine effects of performance enhancers on ileal and cecal microbial communities (MCs). Eight treatment groups included four controls (uninfected-unmedicated [UU], unmedicated-infected, the antibiotic BMD plus the ionophore Coban as positive control, and the ionophore as negative control), and four treatments (probiotics BC-30 and Calsporin; and EO, Crina Poultry Plus, and Crina PoultryAF). Day-old broilers were raised to 14 days in floor pens on used litter and then were moved to Petersime batteries and inoculated at 15 days with mixed Eimeria spp. Ileal and cecal samples were collected at 14 days and 7 days postinfection. Digesta DNA was subjected to pyrosequencing for sequencing of individual cecal bacteria and denaturing gradient gel electrophoresis (DGGE) for determination of changes in ileal and cecal MC according to percentage similarity coefficient (%SC). Pyrosequencing is very sensitive detecting shifts in individual bacterial sequences, whereas DGGE is able to detect gross shifts in entire MC. These combined techniques offer versatility toward identifying feed additive and mild Eimeria infection modulation of broiler MC. Pyrosequencing detected 147 bacterial species sequences. Additionally, pyrosequencing revealed the presence of relatively low levels of the potential human enteropathogens Campylobacter sp. and four Shigella spp. as well as the potential poultry pathogen Clostridiun perfringens. Pre- and postinfection changes in ileal (56%SC) and cecal (78.5%SC) DGGE profiles resulted from the coccidia infection and with increased broiler age. Probiotics and EO changed MC from those seen in UU ilea and ceca. Results potentially reflect the performance enhancement above expectations in comparison to broilers not given the probiotics or the specific EO blends as feed supplements.

Short-term treatment effects produced by the herbst appliance in the mixed dentition

This prospective clinical investigation evaluates the dentoalveolar and skeletal cephalometric changes produced by the Herbst appliance during treatment of mixed dentition patients with Class II division 1 malocclusion. Thirty individuals (15 male and 15 female individuals; initial mean age nine years 10 months) were treated with the Herbst appliance for a period of 12 months. For comparison, the records of 30 untreated Class II children (15 boys, 15 girls; initial mean age nine years eight months) were followed without treatment for a period of 12 months. The results indicated that the treatment effects produced in the mixed dentition patients were primarily dentoalveolar in nature. The mandibular incisors were tipped labially, and the maxillary incisors were retruded; a significant increase in mandibular posterior dentoalveolar height occurred, and there was a restriction in the vertical development of the maxillary molars. There was no difference in the forward growth of the maxilla between the two groups. In comparison with the controls, however, the Herbst treatment produced a modest but statistically significant increase in total mandibular length. This increase in total mandibular length, however, was less than that observed in adolescent Herbst patients in other studies. © 2005 by The EH Angle Education and Research Foundation, Inc.

Changes on facial profile in the mixed dentition, from natural growth and induced by Balters' bionator appliance

Objective: The purpose of this study was to evaluate the facial profile changes induced by Balters' bionator appliance in Class II division 1 patients, at mixed dentition stage. Methods: The sample consisted of 28 prepubertal individuals at stages 1 and 2 of skeletal maturation (CVM), which were divided in two groups. The experimental group consisted of 14 individuals (7 boys and 7 girls, initial mean age of 8y12m) which were treated with Balters' bionator appliance for 14.7 months. The effects of treatment were compared to a control group of 14 subjects (7 boys and 7 girls, initial mean age of 8y5m) with Class II malocclusion, division 1, not orthodontically treated, which were followed up for 15.4 months. The statistical analysis was performed using Student's t test, at a significance level of 5%. Results: The results showed that the Balters' bionator appliance promoted a significant increase on the mentolabial angle, in addition to demonstrating a tendency to reduce the facial skeletal convexity, to restrict the maxillary growth and to increase the nasolabial angle and the lower anterior facial height. Conclusion: It can be concluded that the Balters' bionator appliance improved the facial profile of children treated at mixed dentition stage. © 2013 Dental Press Journal of Orthodontics.

Quantitative genetic study of age at subsequent rebreeding in Nellore cattle by using survival analysis

The continuous trait age at subsequent rebreeding (ASR) was evaluated using survival analysis in Nellore breed cows that conceived for the first time at approximately 14 months of age. This methodology was chosen because the restricted breeding season produces censored data. The dataset contained 2885 records of ASR (in days). Records of females that did not produce calves in the following year after being exposed to a sire were considered censored (48.3% of the total). The statistical model used was a Weibull mixed survival model, which included fixed effects of contemporary groups (CG) and period and a random effect of individual animal. The effect of contemporary groups on ASR was significant (P < 0.01). Heritabilities obtained for ASR were 0.03 and 0.04 in logarithmic and original scales, respectively. These results indicate that the genetic selection response for subsequent reproduction of 2-year-old Nellore breed females is not expected to be effective based on survival analysis. Furthermore, these results suggest that environmental improvement is fundamental to this important trait. It should be highlighted that an increase in the average date of birth can produce an adverse effect in the future, since this cannot be compensated by genetic improvement.

Do Thyroid Ultrasonographic Features Change According to Age in Euthyroid Dogs?

The thyroid gland was assessed by ultrasound in healthy euthyroid mixed-breed medium size dogs in different age groups. The objective was to verify ultrasonographic imaging patterns in these groups, as well as to identify possible changes in imaging features resulting from ageing. Thirty dogs - 10 young (<1 year), 10 adult and 10 elderly - without clinical signs or history of thyroid gland disease with complete blood count and thyroid function tests within the reference values were evaluated. Each thyroid lobe was examined by ultrasound for shape, size, echogenicity and echotexture. The analysis of echogenicity and echotexture was made by histogram. Thyroid volume was estimated by the equation for ellipsoid (length x width x height) pi/6. The thyroid volume of the young dogs in this study had a tendency to be higher than in adult dogs (P = 0.068) and older dogs (P = 0.120). The height of the thyroid lobe in the longitudinal plane was significantly higher (P = 0.026) in young dogs compared with the other dogs. The echotexture and echogenicity had no significant differences between groups, but the echogenicity was greater in older dogs. The results point out that ultrasound imaging of the thyroid volume is influenced by age in euthyroid dogs.

Allometric equations for estimating tree biomass in restored mixed-species Atlantic Forest stands

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The Impact of Teaching Presence in Intensive Online Courses on Perceived Learning and Sense of Community: A Mixed Methods Study

This mixed methods concurrent triangulation design study was predicated upon two models that advocated a connection between teaching presence and perceived learning: the Community of Inquiry Model of Online Learning developed by Garrison, Anderson, and Archer (2000); and the Online Interaction Learning Model by Benbunan-Fich, Hiltz, and Harasim (2005). The objective was to learn how teaching presence impacted students’ perceptions of learning and sense of community in intensive online distance education courses developed and taught by instructors at a regional comprehensive university. In the quantitative phase online surveys collected relevant data from participating students (N = 397) and selected instructional faculty (N = 32) during the second week of a three-week Winter Term. Student information included: demographics such as age, gender, employment status, and distance from campus; perceptions of teaching presence; sense of community; perceived learning; course length; and course type. The students claimed having positive relationships between teaching presence, perceived learning, and sense of community. The instructors showed similar positive relationships with no significant differences when the student and instructor data were compared. The qualitative phase consisted of interviews with 12 instructors who had completed the online survey and replied to all of the open-response questions. The two phases were integrated using a matrix generation, and the analysis allowed for conclusions regarding teaching presence, perceived learning, and sense of community. The findings were equivocal with regard to satisfaction with course length and the relative importance of the teaching presence components. A model was provided depicting relationships between and among teaching presence components, perceived learning, and sense of community in intensive online courses.

Geochronological constraints on the age of a Permo-Triassic impact event: U-Pb and Ar-40/Ar-39 results for the 40 km Araguainha structure of central Brazil

Impact cratering has been a fundamental geological process in Earth history with major ramifications for the biosphere. The complexity of shocked and melted rocks within impact structures presents difficulties for accurate and precise radiogenic isotope age determination, hampering the assessment of the effects of an individual event in the geological record. We demonstrate the utility of a multi-chronometer approach in our study of samples from the 40 km diameter Araguainha impact structure of central Brazil. Samples of uplifted basement granite display abundant evidence of shock deformation, but U/Pb ages of shocked zircons and the Ar-40/Ar-39 ages of feldspar from the granite largely preserve the igneous crystallization and cooling history. Mixed results are obtained from in situ Ar-40/Ar-39 spot analyses of shocked igneous biotites in the granite, with deformation along kink-bands resulting in highly localized, partial resetting in these grains. Likewise, spot analyses of perlitic glass from pseudotachylitic breccia samples reflect a combination of argon inheritance from wall rock material, the age of the glass itself, and post-impact devitrification. The timing of crater formation is better assessed using samples of impact-generated melt rock where isotopic resetting is associated with textural evidence of melting and in situ crystallization. Granular aggregates of neocrystallized zircon form a cluster of ten U-Pb ages that yield a "Concordia" age of 247.8 +/- 3.8 Ma. The possibility of Pb loss from this population suggests that this is a minimum age for the impact event. The best evidence for the age of the impact comes from the U-Th-Pb dating of neocrystallized monazite and Ar-40/Ar-39 step heating of three separate populations of post-impact, inclusion-rich quartz grains that are derived from the infill of miarolitic cavities. The Pb-206/U-238 age of 254.5 +/- 3.2 Ma (2 sigma error) and Pb-208/Th-232 age of 255.2 +/- 4.8 Ma (2 sigma error) of monazite, together with the inverse, 18 point isochron age of 254 +/- 10 Ma (MSWD = 0.52) for the inclusion-rich quartz grains yield a weighted mean age of 254.7 +/- 2.5 Ma (0.99%, 2 sigma error) for the impact event. The age of the Araguainha crater overlaps with the timing of the Permo-Triassic boundary, within error, but the calculated energy released by the Araguainha impact is insufficient to be a direct cause of the global mass extinction. However, the regional effects of the Araguainha impact event in the Parana-Karoo Basin may have been substantial. (C) 2012 Elsevier Ltd. All rights reserved.

Genetic analyses of smoking initiation, persistence, quantity, and age-at-onset of regular cigarette use in Brazilian families: the Baependi Heart Study

Background: The purpose of this study was to estimate the genetic influences on the initiation of cigarette smoking, the persistence, quantity and age-at-onset of regular cigarette use in Brazilian families. Methods: The data set consisted of 1,694 individuals enrolled in the Baependi Heart Study. The heritability and the heterogeneity in genetic and environmental variance components by gender were estimated from variance components approaches, using the SOLAR (Sequential Oligogenic Linkage Analysis Routines) computer package. The mixed-effects Cox model was used for the genetic analysis of the age-at onset of regular cigarette use. Results: The heritability estimates were high (> 50%) for smoking initiation and were intermediate, ranging from 23.4 to 31.9%, for smoking persistence and quantity. Significant evidence for heterogeneity in variance components by gender was observed for smoking initiation and age-at-onset of regular cigarette use. Genetic factors play an important role in the interindividual variation of these phenotypes in females, while in males there is a predominant environmental component, which could be explained by greater social influences in the initiation of tobacco use. Conclusions: Significant heritabilities were observed in smoking phenotypes for both males and females from the Brazilian population. These data add to the literature and are concordant with the notion of significant biological determination in smoking behavior. Samples from the Baependi Heart Study may be valuable for the mapping of genetic loci that modulate this complex biological trait.

Alveolar Bone Morphology in Patients With Bilateral Complete Cleft Lip and Palate in the Mixed Dentition: Cone Beam Computed Tomography Evaluation

Objectives: To verify the thickness and level of alveolar bone around the teeth adjacent to the cleft by means of cone beam computed tomography (CBCT) in patients with complete bilateral cleft lip and palate prior to bone graft surgery and orthodontic intervention. Method: The sample comprised 10 patients with complete bilateral cleft lip and palate (five boys and five girls) in the mixed dentition. The mean age was 9.5 years, and all subjects showed a G3 interarch relationship according to the Bauru index. The thickness of alveolar bone surrounding the maxillary incisors and the maxillary canines was measured in CBCT axial section using the software iCAT Xoran System. The distance between the alveolar bone crest and the cement-enamel junction (CEJ) was measured in cross sections. Results: The tomography images showed a thin alveolar bone plate around teeth adjacent to clefts. No bone dehiscence was observed in teeth adjacent to clefts during the mixed dentition. A slight increase in the distance between the alveolar bone crest and the CEJ was observed in the mesial and lingual aspects of canines adjacent to cleft. Conclusion: In patients with BCLP in the mixed dentition, teeth adjacent to the alveolar cleft are covered by a thin alveolar bone plate. However, the level of alveolar bone crest around these teeth seems to be normal, and no bone dehiscence was identified at this age.

U-Pb zircon in situ dating with LA-MC-ICP-MS using a mixed detector configuration

The LA-MC-ICP-MS method applied to U-Pb in situ dating is still rapidly evolving due to improvements in both lasers and ICP-MS. To test the validity and reproducibility of the method, 5 different zircon samples, including the standard Temora-2, ranging in age between 2.2 Ga and 246 Ma, were dated using both LA-MC-ICP-MS and SHRIMP. The selected zircons were dated by SHRIMP and, after gentle polishing, the laser spot was driven to the same site or on the same zircon phase with a 213 nm laser microprobe coupled to a multi-collector mixed system. The data were collected with a routine spot size of 25 μm and, in some cases, of 15 and 40 μm. A careful cross-calibration using a diluted U-Th-Pb solution to calculate the Faraday reading to counting rate conversion factors and the highly suitable GJ-1 standard zircon for external calibrations were of paramount importance for obtaining reliable results. All age results were concordant within the experimental errors. The assigned age errors using the LA-MC-ICP-MS technique were, in most cases, higher than those obtained by SHRIMP, but if we are not faced with a high resolution stratigraphy, the laser technique has certain advantages.

Progression of age-related geographic atrophy: role of the fellow eye

PURPOSE. To evaluate the role of fellow eye status in determining progression of geographic atrophy (GA) in patients with age-related macular degeneration (AMD). METHODS. A total of 300 eyes with GA of 193 patients from the prospective, longitudinal, natural history FAM Study were classified into three groups according to the AMD manifestation in the fellow eye at baseline examination: (1) bilateral GA, (2) early/intermediate AMD, and (3) exudative AMD. GA areas were quantified based on fundus autofluorescence images using a semiautomated image-processing method, and progression rates (PR) were estimated using two-level, linear, mixed-effects models. RESULTS. Crude GA-PR in the bilateral GA group (mean, 1.64 mm(2)/y; 95% CI, 1.478-1.803) was significantly higher than in the fellow eye early/intermediate group (0.74 mm(2)/y, 0.146-1.342). Although there was a significant difference in baseline GA size (P = 0.0013, t-test), and there was a significant increase in GA-PR by 0.11 mm(2)/y (0.05-0.17) per 1 disc area (DA; 2.54 mm(2)), an additional mean change of -0.79 (-1.43 to -0.15) was given to the PR beside the effect of baseline GA size. However, this difference was only significant when GA size was ?1 DA at baseline with a GA-PR of 1.70 mm(2)/y (1.54-1.85) in the bilateral and 0.95 mm(2)/y (0.37-1.54) in the early/intermediate group. There was no significant difference in PR compared with that in the fellow eye exudative group. CONCLUSIONS. The results indicate that the AMD manifestation of the fellow eye at baseline serves as an indicator for disease progression in eyes with GA ? 1 DA. Predictive characteristics not only contribute to the understanding of pathophysiological mechanisms, but also are useful for the design of future interventional trials in GA patients.

A natural freshwater origin for two chlamydial species, Candidatus Piscichlamydia salmonis and Candidatus Clavochlamydia salmonicola, causing mixed infections in wild brown trout (Salmo trutta)

Gill disease in salmonids is characterized by a multifactorial aetiology. Epitheliocystis of the gill lamellae caused by obligate intracellular bacteria of the order Chlamydiales is one known factor; however, their diversity has greatly complicated analyses to establish a causal relationship. In addition, tracing infections to a potential environmental source is currently impossible. In this study, we address these questions by investigating a wild brown trout (Salmo trutta) population from seven different sites within a Swiss river system. One age class of fish was followed over 18 months. Epitheliocystis occurred in a site-specific pattern, associated with peak water temperatures during summer months. No evidence of a persistent infection was found within the brown trout population, implying an as yet unknown environmental source. For the first time, we detected 'Candidatus Piscichlamydia salmonis' and 'Candidatus Clavochlamydia salmonicola' infections in the same salmonid population, including dual infections within the same fish. These organisms are strongly implicated in gill disease of caged Atlantic salmon in Norway and Ireland. The absence of aquaculture production within this river system and the distance from the sea, suggests a freshwater origin for both these bacteria and offers new possibilities to explore their ecology free from aquaculture influences.

Mother-Child Spiritual Discourse: A Mixed Methods Study

Parents and children, starting at very young ages, discuss religious and spiritual issues¿where we come from, what happens to us after we die, is there a God, and so on. Unfortunately, few studies have analyzed the content and structure of parent-child conversation about religion and spirituality (Boyatzis & Janicki, 2003; Dollahite & Thatcher, 2009), and most studies have relied on self-report with no direct observation. The current study examined mother-child (M-C) spiritual discourse to learn about its content, structure, and frequency through a survey inventory in combination with direct video observation using a novel structured task. We also analyzed how mothers¿ religiosity along several major dimensions related to their communication behaviors within both methods. Mothers (N = 39, M age = 40) of children aged 3-12 completed a survey packet on M-C spiritual discourse and standard measures of mothers¿ religious fundamentalism, intrinsic religiosity, sanctification of parenting (how much the mother saw herself as doing God¿s work as a parent), and a new measure of parental openness to children¿s spirituality. Then, in a structured task in our lab, mothers (N = 33) and children (M age = 7.33) watched a short film or read a short book that explored death in an age-appropriate manner and then engaged in a videotaped conversation about the movie or book and their religious or spiritual beliefs. Frequency of M-C spiritual discourse was positively related to mothers¿ religious fundamentalism (r = .71, p = .00), intrinsic religiosity (r = .77, p = .00), and sanctification of parenting (r = .79, p = .00), but, surprisingly, was inversely related to mothers¿ v openness to child¿s spirituality (r = -.52, p = .00). Survey data showed that the two most common topics discussed were God (once a week) and religion as it relates to moral issues (once a week). According to mothers their children¿s most common method of initiating spiritual discourse was to repeat what he or she has heard parents or family say about religious issues (M = 2.97; once a week); mothers¿ most common method was to describe their own religious/spiritual beliefs (M = 2.92). Spiritual discourse most commonly occurred either at bedtime or mealtime as reported by 26% of mothers, with the most common triggers reported as daily routine/random thoughts (once a week) and observations of nature (once a week). Mothers¿ most important goals for spiritual discourse were to let their children know that they love them (M = 3.72; very important) and to help them become a good and moral person (M = 3.67; very important). A regression model showed that significant variance in frequency of mother-child spiritual discourse (R2 = .84, p = .00) was predicted by the mothers¿ importance of goals during discourse (ß = 0.46, p = .00), frequency that the mother¿s spirituality was deepened through spiritual discourse (ß = 0.39, p = .00), and the mother¿s fundamentalism (ß = 0.20, p = .05). In a separate regression, the mother¿s comfort in the structured task (ß = 0.70, p = .00), and the number of open-ended questions she asked (ß = -0.26, p = .03) predicted the reciprocity between mother and child (R2 = .62, p = .00). In addition, the mother¿s age (ß = 0.22, p = .059) and comfort during the task (ß = 0.73, p = .00) predicted the child¿s engagement within the structured task. Other findings and theoretical and methodological implications will be discussed.

A Spanish founder mutation in the chloride channel gene, CLCNKB, as a cause of atypical Bartter syndrome in adult age

BACKGROUND: Mutations in the chloride channel gene, CLCNKB, usually cause classic Bartter syndrome (cBS) or a mixed Bartter-Gitelman phenotype in the first years of life. METHODS: We report an adult woman with atypical BS caused by a homozygous missense mutation, A204T, in the CLCNKB gene, which has previously been described as the apparently unique cause of cBS in Spain. RESULTS: The evaluation of this patient revealed an overlap of phenotypic features ranging from severe biochemical and systemic disturbances typical of cBS to scarce symptoms and diagnosis in the adult age typical of Gitelman syndrome. The tubular disease caused a dramatic effect on mental, growth and puberal development leading to low IQ, final short stature and abnormal ovarian function. Furthermore, low serum PTH concentrations with concomitant nephrocalcinosis and normocalcaemia were observed. Both ovarian function and serum PTH levels were normalized after treatment with cyclooxygenase inhibitors. CONCLUSIONS: The present report confirms a weak genotype-phenotype correlation in patients with CLCNKB mutations and supports the founder effect of the A204T mutation in Spain. In our country, the genetic diagnosis of adult patients with hereditary hypokalaemic tubulopathies should include a screening of A204T mutation in the CLCNKB gene.