Genotypic Characterization of Fungal Species Isolated From Broiler Breeder Chickens, Dead-In-Shell and Hatched Chicks

To investigate the prevalence of fungal agents in local Egyptian broiler breeder chicken’s premises, tracheal and cloacal swabs from chickens, feed, and water samples were collected. The targeted breed's dead-in-shell eggs and newly hatched chick’s samples were also tested. All fungal isolates were morphologically typed and the predominant fungal species were further subjected to molecular typing using PCR-RFLP and gene sequence analysis of the β-tubulin gene. Results revealed a high prevalence of fungal isolates in tracheal and cloacal swabs (39.3 - 48.1%) and feed and water samples (37.5% and 28.6%, respectively). Fungal isolation rates in dead-in-shell eggs of all breeds were high except in Dahaby breed. Aspergillus species including A. niger, A. flavus, and A. terreus were the predominantly isolated fungi from all collected samples. The ß-Tubulin genes PCR-RFLP of selected Aspergillus isolates showed a characteristic restriction pattern for each species; however, the method was unable to distinguish between strains. The β-tubulin gene phylogenetic and sequence analysis of selected A. flavus, and A. terreus from breeder chickens and their hatching chicks indicated their relatedness to isolates from bronchopulmonary Aspergillosis in humans in the Middle East. In conclusion, the Aspergillus species remains the most prevalent fungi in breeder chickens, their incubated eggs and hatched chicks indicating their widespread in hatcheries. The PCR-RFLP is an easy tool to discriminate between Aspergilli species, however, the β-tubulin sequence analysis more descriptive of potential sources of fungal contamination. Further epidemiological studies are needed to monitor avian and human Aspergilli in poultry houses with a special focus on antifungal drug-resistant strains

Long-term perturbations due to a disturbing body in elliptic inclined orbit

In the current study, a double-averaged analytical model including the action of the perturbing body's inclination is developed to study third-body perturbations. The disturbing function is expanded in the form of Legendre polynomials truncated up to the second-order term, and then is averaged over the periods of the spacecraft and the perturbing body. The efficiency of the double-averaged algorithm is verified with the full elliptic restricted three-body model. Comparisons with the previous study for a lunar satellite perturbed by Earth are presented to measure the effect of the perturbing body's inclination, and illustrate that the lunar obliquity with the value 6.68\degree is important for the mean motion of a lunar satellite. The application to the Mars-Sun system is shown to prove the validity of the double-averaged model. It can be seen that the algorithm is effective to predict the long-term behavior of a high-altitude Martian spacecraft perturbed by Sun. The double-averaged model presented in this paper is also applicable to other celestial systems.Comment: 28 pages, 6 figure

Relative Effectiveness of Mating Success and Sperm Competition at Eliminating Deleterious Mutations in Drosophila melanogaster

Condition-dependence theory predicts that sexual selection will facilitate adaptation by selecting against deleterious mutations that affect the expression of sexually selected traits indirectly via condition. Recent empirical studies have provided support for this prediction; however, their results do not elucidate the relative effects of pre- and postcopulatory sexual selection on deleterious mutations. We used the Drosophila melanogaster model system to discern the relative contributions of pre- and postcopulatory processes to selection against deleterious mutations. To assess second-male ejaculate competition success (P2; measured as the proportion of offspring attributable to the experimental male) and mating success, mutant and wild-type male D. melanogaster were given the opportunity to mate with females that were previously mated to a standard competitor male. This process was repeated for males subjected to a diet quality manipulation to test for effects of environmentally-manipulated condition on P2 and mating success. While none of the tested mutations affected P2, there was a clear effect of condition. Conversely, several of the mutations affected mating success, while condition showed no effect. Our results suggest that precopulatory selection may be more effective than postcopulatory selection at removing deleterious mutations. The opposite result obtained for our diet manipulation points to an interesting discrepancy between environmental and genetic manipulations of condition, which may be explained by the multidimensionality of condition. Establishing whether the various stages of sexual selection affect deleterious mutations differently, and to what extent, remains an important issue to resolve

Electrochemical determination of microRNAs based on isothermal strand-displacement polymerase reaction coupled with multienzyme functionalized magnetic micro-carriers

This study was supported by the National Natural Science Foundation of China (81371901), Doctoral Scientific Fund Project of the Ministry of Education of People's Republic of China (20134433110010), the Critical Point-of-Care Testing (CPOCT) Research grant of American Association for Clinical Chemistry (AACC) and 2015 Distinguished Academic Fellowships of Royal College of Engineering (DVF1415/2/79)

Utilisation of an operative difficulty grading scale for laparoscopic cholecystectomy

Background A reliable system for grading operative difficulty of laparoscopic cholecystectomy would standardise description of findings and reporting of outcomes. The aim of this study was to validate a difficulty grading system (Nassar scale), testing its applicability and consistency in two large prospective datasets. Methods Patient and disease-related variables and 30-day outcomes were identified in two prospective cholecystectomy databases: the multi-centre prospective cohort of 8820 patients from the recent CholeS Study and the single-surgeon series containing 4089 patients. Operative data and patient outcomes were correlated with Nassar operative difficultly scale, using Kendall’s tau for dichotomous variables, or Jonckheere–Terpstra tests for continuous variables. A ROC curve analysis was performed, to quantify the predictive accuracy of the scale for each outcome, with continuous outcomes dichotomised, prior to analysis. Results A higher operative difficulty grade was consistently associated with worse outcomes for the patients in both the reference and CholeS cohorts. The median length of stay increased from 0 to 4 days, and the 30-day complication rate from 7.6 to 24.4% as the difficulty grade increased from 1 to 4/5 (both p < 0.001). In the CholeS cohort, a higher difficulty grade was found to be most strongly associated with conversion to open and 30-day mortality (AUROC = 0.903, 0.822, respectively). On multivariable analysis, the Nassar operative difficultly scale was found to be a significant independent predictor of operative duration, conversion to open surgery, 30-day complications and 30-day reintervention (all p < 0.001). Conclusion We have shown that an operative difficulty scale can standardise the description of operative findings by multiple grades of surgeons to facilitate audit, training assessment and research. It provides a tool for reporting operative findings, disease severity and technical difficulty and can be utilised in future research to reliably compare outcomes according to case mix and intra-operative difficulty

Human breast cancer-derived soluble factors facilitate CCL19-induced chemotaxis of human dendritic cells

Breast cancer remains as a challenging disease with high mortality in women. Increasing evidence points the importance of understanding a crosstalk between breast cancers and immune cells, but little is known about the effect of breast cancer-derived factors on the migratory properties of dendritic cells (DCs) and their consequent capability in inducing T cell immune responses. Utilizing a unique 3D microfluidic device, we here showed that breast cancers (MCF-7, MDA-MB-231, MDA-MB-436 and SK-BR-3)-derived soluble factors increase the migration of DCs toward CCL19. The enhanced migration of DCs was mainly mediated via the highly activated JNK/c-Jun signaling pathway, increasing their directional persistence, while the velocity of DCs was not influenced, particularly when they were co-cultured with triple negative breast cancer cells (TNBCs or MDA-MB-231 and MDA-MB-436). The DCs up-regulated inflammatory cytokines IL-1?? and IL-6 and induced T cells more proliferative and resistant against activation-induced cell death (AICD), which secret high levels of inflammatory cytokines IL-1??, IL-6 and IFN-??. This study demonstrated new possible evasion strategy of TNBCs utilizing their soluble factors that exploit the directionality of DCs toward chemokine responses, leading to the building of inflammatory milieu which may support their own growth.ope

A four-kallikrein panel for the prediction of repeat prostate biopsy: data from the European Randomized Study of Prostate Cancer Screening in Rotterdam, Netherlands

Background: Most men with elevated levels of prostate-specific antigen (PSA) do not have prostate cancer, leading to a large number of unnecessary biopsies. A statistical model based on a panel of four kallikreins has been shown to predict the outcome of a first prostate biopsy. In this study, we apply the model to an independent data set of men with previous negative biopsy but persistently elevated PSA. Methods: The study cohort consisted of 925 men with a previous negative prostate biopsy and elevated PSA (≥3 ng ml-1), with 110 prostate cancers detected (12%). A previously published statistical model was applied, with recalibration to reflect the lower positive biopsy rates on rebiopsy. Results: The full-kallikrein panel had higher discriminative accuracy than PSA and DRE alone, with area under the curve (AUC) improving from 0.58 (95% confidence interval (CI): 0.52, 0.64) to 0.68 (95% CI: 0.62, 0.74), P<0.001, and high-grade cancer (Gleason 7) at biopsy with AUC improving from 0.76 (95% CI: 0.64, 0.89) to 0.87 (95% CI: 0.81, 0.94), P<0.003). Application of the panel to 1000 men with persistently elevated PSA after initial negative biopsy, at a 15% risk threshold would reduce the number of biopsies by 712; would miss (or delay) the diagnosis of 53 cancers, of which only 3 would be Gleason 7 and the rest Gleason 6 or less. Conclusions: Our data constitute an external validation of a previously published model. The four-kallikrein panel predicts the result of repeat prostate biopsy in men with elevated PSA while dramatically decreasing unnecessary biopsies