TMPRSS2-ERG Fusions Are Strongly Linked to Young Patient Age in Low-grade Prostate Cancer

Based on next-generation sequencing of early-onset prostate cancer (PCa), we earlier demonstrated that PCa in young patients is prone to rearrangements involving androgen-regulated genes—such as transmembrane protease, serine 2 (TMPRSS2)–v-ets avian erythroblastosis virus E26 oncogene homolog (ERG) fusion—and provided data suggesting that this situation might be caused by increased androgen signaling in younger men. In the same study, an accumulation of chromosomal deletions was found in cancers of elderly patients. To determine how age-dependent molecular features relate to cancer phenotype, an existing data set of 11 152 PCas was expanded by additional fluorescence in situ hybridization analyses of phosphatase and tensin homolog (PTEN), 6q15 and 5q21. The results demonstrate that the decrease in TMPRSS2–ERG fusions with increasing patient age is limited to low-grade cancers (Gleason ≤3 + 4) and that the significant increase in the deletion frequency with age was strictly limited to ERG-negative cancers for 6q15 and 5q21 but to ERG-positive cancers for PTEN. These data suggest that the accumulation of non–androgen-linked genomic alterations with advanced patient age may require an appropriate microenvironment, such as a positive or negative ERG status. The strong link of ERG activation to young patient age and low-grade cancers may help to explain a slight predominance of low-grade cancers in young patients.     

A new and automated method for objective analysis of detrusor rhythm during the filling phase

Introduction

There is growing acceptance that the detrusor muscle is not silent during the filling phase of the micturition cycle but displays low-amplitude phasic contractions that have been associated with urinary urgency. Unfortunately, there is currently no standardized methodology to quantify detrusor rhythm during the filling phase. Therefore, the purpose of this study was to develop an automated computer algorithm to analyze rat detrusor rhythm in a quick, accurate, and reproducible manner.

Materials and methods

Strips of detrusor smooth muscle from rats (n = 17) were placed on force transducers and subjected to escalating doses of PGE₂ to generate contractile rhythm tracings. An automated computer algorithm was developed to analyze contractile frequency, amplitude, and tone on the generated rhythm tracings. Results of the automated computerized analysis were compared to human (n = 3) interpretations. Human interpreters manually counted contractions and then recounted the same data two weeks later. Intra-observer, inter-observer, and human-to-computer comparisons were performed.

Results

The computer algorithm quantified concentration-dependent changes in contractile frequency, amplitude, and tone after administration of PGE₂ (10^?9–10^?6M). Concentration–response curves were similar for all contractile components with increases in frequency identified mainly at physiologic concentrations of PGE₂ and increases in amplitude at supra-physiologic concentrations. The computer algorithm consistently over-counted the human interpreters, but with less variability. Differences in inter-observer consistency were statistically significant.

Conclusions

Our computerized algorithm accurately and consistently identified changes in detrusor muscle contractile frequency, amplitude, and tone with varying doses of PGE₂. Frequency counts were consistently higher than those obtained by human interpreters but without variability or bias. Refinements of this method may allow for more standardized approach in the study of pharmacologic agents on filling phase rhythmic activity.     

High-resolution photoacoustic tomography of resting-state functional connectivity in the mouse brain

The increasing use of mouse models for human brain disease studies presents an emerging need for a new functional imaging modality. Using optical excitation and acoustic detection, we developed a functional connectivity photoacoustic tomography system, which allows noninvasive imaging of resting-state functional connectivity in the mouse brain, with a large field of view and a high spatial resolution. Bilateral correlations were observed in eight functional regions, including the olfactory bulb, limbic, parietal, somatosensory, retrosplenial, visual, motor, and temporal regions, as well as in several subregions. The borders and locations of these regions agreed well with the Paxinos mouse brain atlas. By subjecting the mouse to alternating hyperoxic and hypoxic conditions, strong and weak functional connectivities were observed, respectively. In addition to connectivity images, vascular images were simultaneously acquired. These studies show that functional connectivity photoacoustic tomography is a promising, noninvasive technique for functional imaging of the mouse brain.Resting-state functional connectivity (RSFC) is an emerging neuroimaging approach that aims to identify low-frequency, spontaneous cerebral hemodynamic fluctuations and their associated functional connections (1, 2). Recent research suggests that these fluctuations are highly correlated with local neuronal activity (3, 4). The spontaneous fluctuations relate to activity that is intrinsically generated by the brain, instead of activity attributable to specific tasks or stimuli (2). A hallmark of functional organization in the cortex is the striking bilateral symmetry of corresponding functional regions in the left and right hemispheres (5). This symmetry also exists in spontaneous resting-state hemodynamics, where strong correlations are found interhemispherically between bilaterally homologous regions as well as intrahemispherically within the same functional regions (3). Clinical studies have demonstrated that RSFC is altered in brain disorders such as stroke, Alzheimer’s disease, schizophrenia, multiple sclerosis, autism, and epilepsy (6–12). These diseases disrupt the healthy functional network patterns, most often reducing correlations between functional regions. Due to its task-free nature, RSFC imaging requires neither stimulation of the subject nor performance of a task during imaging (13). Thus, it can be performed on patients under anesthesia (14), on patients unable to perform cognitive tasks (15, 16), and even on patients with brain injury (17, 18).RSFC imaging is also an appealing technique for studying brain diseases in animal models, in particular the mouse, a species that holds the largest variety of neurological disease models (3, 13, 19, 20). Compared with clinical studies, imaging genetically modified mice allows exploration of molecular pathways underlying the pathogenesis of neurological disorders (21). The connection between RSFC maps and neurological disorders permits testing and validation of new therapeutic approaches. However, conventional neuroimaging modalities cannot easily be applied to mice. For instance, in functional connectivity magnetic resonance imaging (fcMRI) (22), the resting-state brain activity is determined via the blood-oxygen-level–dependent (BOLD) signal contrast, which originates mainly from deoxy-hemoglobin (23). The correlation analysis central to functional connectivity requires a high signal-to-noise ratio (SNR). However, achieving a sufficient SNR is made challenging by the high magnetic fields and small voxel size needed for imaging the mouse brain, as well as the complexity of compensating for field inhomogeneities caused by tissue–bone or tissue–air boundaries (24). Functional connectivity mapping with optical intrinsic signal imaging (fcOIS) was recently introduced as an alternative method to image functional connectivity in mice (3, 20). In fcOIS, changes in hemoglobin concentrations are determined based on changes in the reflected light intensity from the surface of the brain (3, 25). Therefore, neuronal activity can be measured through the neurovascular response, similar to the method used in fcMRI. However, due to the diffusion of light in tissue, the spatial resolution of fcOIS is limited, and experiments have thus far been performed using an exposed skull preparation, which increases the complexity for longitudinal imaging.Photoacoustic imaging of the brain is based on the acoustic detection of optical absorption from tissue chromophores, such as oxy-hemoglobin (HbO₂) and deoxy-hemoglobin (Hb) (26, 27). This imaging modality can simultaneously provide high-resolution images of the brain vasculature and hemodynamics with intact scalp (28, 29). In this article, we perform functional connectivity photoacoustic tomography (fcPAT) to study RSFC in live mice under either hyperoxic or hypoxic conditions, as well as in dead mice. Our experiments show that fcPAT is able to detect connectivities between different functional regions and even between subregions, promising a powerful functional imaging modality for future brain research.     

Intraoperative Fracture During Staged Total Knee Reimplantation in the Treatment of Periprosthetic Infection

Bone stock during knee reimplantation for infection is compromised and may contribute to intraoperative fracture. This study aims to describe the prevalence of said fractures. A retrospective review was performed of patients who underwent a staged TKA reimplantation for a periprosthetic infection. Patients who sustained an intraoperative fracture were analyzed. The fracture timing, location, and treatment were recorded. Fracture healing, component stability, and need for re-revision were noted. Between 1990 and 2010, 894 reimplantations were performed. Twenty-three fractures occurred in 21 patients (2.3%) with mean follow-up of 56 months (range: 4–122). Thirteen fractures occurred in femora, 7 in tibiae, and 3 in patellae. Four occurred during resection, while 19 occurred during reimplantation. Observation and wires/cables were the most common treatments utilized. At final follow-up, 91% of fractures demonstrated union and 75% of patients demonstrated stable components. Eight patients (38%) required a revision, the majority of which were performed for re-infection.