Balanoposthitis, dyspareunia, and treponema in the critically endangered Gilbert's potoroo (Potorous gilbertii)

The Gilbert's potoroo (Potorous gilbertii) is one of Australia's most critically endangered mammals with a current estimated population of 70 individuals. Both the wild and captive populations have a long history of balanoposthitis with associated crusting, ulceration, and preputial discharge. We sought to identify the microbial species found in the discharge, determine their significance in causing balanoposthitis, and correlate these findings with reproductive success and survivorship. Bacteriologic examination revealed the discharge to be a polymicrobial infection involving Treponema spp., Actinobacillus spp., and Pasteurella spp. Preputial histopathology reported a moderate, chronic, erosive inflammatory response with diffuse, moderate to marked secondary epithelial hyperplasia in conjunction with moderate numbers of spirochetes, suggesting a causative relationship. Clinical examination, preputial biopsies, and serologic screening found no evidence of associated systemic disease. The clinical investigation of Treponema is significant with respect to the overall recovery of Gilbert's potoroo, given the clinical and histopathologic similarities to Treponema paraluis-cuniculi found in rabbits, causing dyspareunia, and the severity of the associated balanoposthitis.     

The influence of CpG and UpA dinucleotide frequencies on RNA virus replication and characterization of the innate cellular pathways underlying virus attenuation and enhanced replication

Most RNA viruses infecting mammals and other vertebrates show profound suppression of CpG and UpA dinucleotide frequencies. To investigate this functionally, mutants of the picornavirus, echovirus 7 (E7), were constructed with altered CpG and UpA compositions in two 1.1–1.3 Kbase regions. Those with increased frequencies of CpG and UpA showed impaired replication kinetics and higher RNA/infectivity ratios compared with wild-type virus. Remarkably, mutants with CpGs and UpAs removed showed enhanced replication, larger plaques and rapidly outcompeted wild-type virus on co-infections. Luciferase-expressing E7 sub-genomic replicons with CpGs and UpAs removed from the reporter gene showed 100-fold greater luminescence. E7 and mutants were equivalently sensitive to exogenously added interferon-β, showed no evidence for differential recognition by ADAR1 or pattern recognition receptors RIG-I, MDA5 or PKR. However, kinase inhibitors roscovitine and C16 partially or entirely reversed the attenuated phenotype of high CpG and UpA mutants, potentially through inhibition of currently uncharacterized pattern recognition receptors that respond to RNA composition. Generating viruses with enhanced replication kinetics has applications in vaccine production and reporter gene construction. More fundamentally, the findings introduce a new evolutionary paradigm where dinucleotide composition of viral genomes is subjected to selection pressures independently of coding capacity and profoundly influences host–pathogen interactions.     

The Added Diagnostic Value of Dynamic Contrast-Enhanced MRI at 3.0 T in Nonpalpable Breast Lesions

Objective

To investigate the added diagnostic value of 3.0 Tesla breast MRI over conventional breast imaging in the diagnosis of in situ and invasive breast cancer and to explore the role of routine versus expert reading.

Materials and Methods

We evaluated MRI scans of patients with nonpalpable BI-RADS 3–5 lesions who underwent dynamic contrast-enhanced 3.0 Tesla breast MRI. Initially, MRI scans were read by radiologists in a routine clinical setting. All histologically confirmed index lesions were re-evaluated by two dedicated breast radiologists. Sensitivity and specificity for the three MRI readings were determined, and the diagnostic value of breast MRI in addition to conventional imaging was assessed. Interobserver reliability between the three readings was evaluated.

Results

MRI examinations of 207 patients were analyzed. Seventy-eight of 207 (37.7%) patients had a malignant lesion, of which 33 (42.3%) patients had pure DCIS and 45 (57.7%) invasive breast cancer. Sensitivity of breast MRI was 66.7% during routine, and 89.3% and 94.7% during expert reading. Specificity was 77.5% in the routine setting, and 61.0% and 33.3% during expert reading. In the routine setting, MRI provided additional diagnostic information over clinical information and conventional imaging, as the Area Under the ROC Curve increased from 0.76 to 0.81. Expert MRI reading was associated with a stronger improvement of the AUC to 0.87. Interobserver reliability between the three MRI readings was fair and moderate.

Conclusions

3.0 T breast MRI of nonpalpable breast lesions is of added diagnostic value for the diagnosis of in situ and invasive breast cancer.     

Sex‐ And tissue‐specific differences in telomere length in a reptile

The usage of telomere length (TL) in blood as a proxy for the TL of other tissues relies on the assumption that telomere dynamics across all tissues are similar. However, telomere attrition can be caused by reactive oxygen species (ROS) which may vary with metabolic rate, which itself varies across organs depending upon the life history strategy of an organism. Thus, we chose to measure the telomeres of various cell types in juvenile painted dragon lizards, Ctenophorus pictus, given their unusual life history strategy. Individuals typically only experience a single mating season. We measured the TL of male and female dragons using qPCR and observed that TL varied with tissue type and sex. Telomeres of blood cells were longer than those of liver, heart, brain, and spleen, and females had longer telomeres than males. Brain telomeres in males were approximately half the length of those in females. Telomeric attrition in the male brain may be due to the need for rapid learning of reproductive tactics (territory patrol and defense, mate‐finding). Significant correlations between the TL of tissue types suggest that blood TL may be a useful proxy for the TL of other tissues. Our comparison of organ‐specific telomere dynamics, the first in a reptile, suggests that the usage of blood TL as a proxy requires careful consideration of the life history strategy of the organism.     

SH-SY5Y and LUHMES cells display differential sensitivity to MPP+, tunicamycin,and epoxomicin in 2D and 3D cell culture

SH-SY5Y and LUHMES cell lines are widely used as model systems for studying neurotoxicity. Most of the existing data regarding the sensitivity of these cell lines to neurotoxicants have been recorded from cells growing as two-dimensional (2D) cultures on the surface of glass or plastic. With the emergence of 3D culture platforms designed to better represent native tissue, there is a growing need to compare the toxicology of neurons grown in 3D environments to those grown in 2D to better understand the impact that culture environment has on toxicant sensitivity. Here, a simple 3D culture method was used to assess the impact of growth environment on the sensitivity of SH-SY5Y cells and LUHMES cells to MPP+, tunicamycin, and epoxomicin, three neurotoxicants that have been previously used to generate experimental models for studying Parkinson's disease pathogenesis. SH-SY5Y cell viability following treatment with these three toxicants was significantly lower in 2D cultures as compared to 3D cultures. On the contrary, LUHMES cells did not show significant differences between growth conditions for any of the toxicants examined. However, LUHMES cells were more sensitive to MPP+, tunicamycin, and epoxomicin than SH-SY5Y cells. Thus, both the choice of cell line and the choice of growth environment must be considered when interpreting in vitro neurotoxicity data.     

DNA methylation changes in atypical adenomatous hyperplasia, adenocarcinoma in situ, and lung adenocarcinoma

Background

Aberrant DNA methylation is common in lung adenocarcinoma, but its timing in the phases of tumor development is largely unknown. Delineating when abnormal DNA methylation arises may provide insight into the natural history of lung adenocarcinoma and the role that DNA methylation alterations play in tumor formation.

Methodology/Principal Findings

We used MethyLight, a sensitive real-time PCR-based quantitative method, to analyze DNA methylation levels at 15 CpG islands that are frequently methylated in lung adenocarcinoma and that we had flagged as potential markers for non-invasive detection. We also used two repeat probes as indicators of global DNA hypomethylation. We examined DNA methylation in 249 tissue samples from 93 subjects, spanning the putative spectrum of peripheral lung adenocarcinoma development: histologically normal adjacent non-tumor lung, atypical adenomatous hyperplasia (AAH), adenocarcinoma in situ (AIS, formerly known as bronchioloalveolar carcinoma), and invasive lung adenocarcinoma. Comparison of DNA methylation levels between the lesion types suggests that DNA hypermethylation of distinct loci occurs at different time points during the development of lung adenocarcinoma. DNA methylation at CDKN2A ex2 and PTPRN2 is already significantly elevated in AAH, while CpG islands at 2C35, EYA4, HOXA1, HOXA11, NEUROD1, NEUROD2 and TMEFF2 are significantly hypermethylated in AIS. In contrast, hypermethylation at CDH13, CDX2, OPCML, RASSF1, SFRP1 and TWIST1 and global DNA hypomethylation appear to be present predominantly in invasive cancer.

Conclusions/Significance

The gradual increase in DNA methylation seen for numerous loci in progressively more transformed lesions supports the model in which AAH and AIS are sequential stages in the development of lung adenocarcinoma. The demarcation of DNA methylation changes characteristic for AAH, AIS and adenocarcinoma begins to lay out a possible roadmap for aberrant DNA methylation events in tumor development. In addition, it identifies which DNA methylation changes might be used as molecular markers for the detection of preinvasive lesions.     

Karyotype diversity and fish conservation of southern field from South Brazil

Six hundred and twenty-seven specimens of neotropical fishes belonging to thirteen species/populations from southern field (Paraná, State, Brazil) were karyotypicaly analyzed between 2000 and 2006. The study biogeographic region is characterized as a divisor of waters presenting tributaries and headboards of the hydrographic basins of the rivers Tibagi, Iguaçu, Ivaí, and Paranapanema (Upper Paraná Basin) and Ribeira river (East Basin). Two patterns of karyotypic diversification were verified, being one more conserved in the karyotypic macrostructure and the other more diversified. The understanding at of this variability with views to the citotaxonomy and biological conservation was interpreted at the light of (1) the historical biogeography, (2) of the evolutionary time, and (3) of the biology of the species, indicating the study area as highly relevant for the selection of evolutionary potential.     

Chromosomal differentiation and speciation in sister-species of Grammatidae (Perciformes) from the Western Atlantic

In the tropical Atlantic, the ichthyofauna between the coast of Brazil and the Caribbean regions, divided by the Amazon barrier, is very similar presenting several geminate species, including Gramma brasiliensis, endemic in Brazil, and its Caribbean counterpart Gramma loreto. Morphological and molecular studies have helped establish evolutionary patterns that sister-species of these two marine habitats are subjected to. However, their chromosomal characteristics are only beginning to be better characterized. Accordingly, a comparative cytogenetic analysis was carried out in G. brasiliensis and G. loreto, seeking evidence of cytotaxonomic markers implicated in the karyotypic diversification of these species and likely associated with speciation events. Heterochromatic regions and their affinity to fluorochromes GC- or AT-specific were identified, as well as the distribution of ribosomal DNA sites in chromosomes, either by silver nitrate impregnation (Ag-NORs) or dual-color FISH mapping with 18S and 5S rDNA probes. While displaying the same diploid number, 2n?=?48 chromosomes, considered basal for Perciformes, the two species differed in karyotype structure, showing karyotypic formulas and species-specific heterochromatin pattern. The cytological characters found support the differentiating status of these species, possibly achieved under the conditions of allopatry due to the Amazon/Orinoco barrier, showing chromosomal peculiarities in Grammatidae species when compared to other groups of Perciformes.