Analgesia and Humane Endpoints for Rodents in Sepsis Research.

Numerous regulatory bodies around the world require analgesics for rodents undergoing surgery to induce sepsis. Well-controlled pain will decrease morbidity. Options for analgesics include NSAIDs, local analgesics, and opioids. Supportive care can also decrease stress to post-operative animals. As well, humane endpoints should be agreed upon before the study commences so as to alleviate unnecessary pain and distress.

Comparison of transperineal ultrasound image guidance technique to transabdominal technique for prostate radiation therapy.

INTRODUCTION: Ultrasound (US) guidance of the prostate has long been conducted using a transabdominal (TA) approach. More recently, a transperineal (TP) approach has been made available for image guidance. Our aim was to determine if both methods produced similar alignments within the same patients. MATERIALS AND METHODS: We utilized two clinical US image guidance (IG) systems (Elekta Clarity and Best BAT). The B-mode Acquisition and Targeting USIG system is a bi-planar, so-called 2.5D USIG system, that is acquired TA. Clarity is a 3D US system that generates a volumetric 3D US data set and US-derived IG contours that are coregistered to the planning CT images. The probe is oriented in the sagittal plane against the perineum (TP). After positioning the patient for treatment using the TP USIG, we maintained the position defined by Clarity tracking and then acquired a TA-based USIG. The two US-based methods of localizing the prostate (TA vs TP) were compared via Bland-Altman (BA) statistical analysis to determine if there was alignment agreement between methods. RESULTS: The BA test for all 101 patients, 2093 fractions resulted in 95% confidence intervals (upper and lower limits of the BA test) of 0.6 mm in LR, 0.9 mm in AP and 1.0 mm in SI. The bias between the two systems was calculated as 0.03, 0.02, and 0.03 mm in LR, AP, and SI. CONCLUSIONS: Both systems resulted in statistically equivalent targeting positions for the prostate. Because of the unique intrafraction, real-time motion tracking capability of the TP system, this solution represents a unique extension to the previously reported clinical benefits of a TA approach by providing assurance of the prostate remaining in the treatment field during beam-on.

Effects of Various Commercially Available Enrichment Options on Handling and Chronic Stress Markers in Female ICR Mice.

Although social housing of mice generally is preferred, mice must be individually housed in some situations. In these cases, enhanced attention to environmental enrichment is encouraged, but few studies assess the wellbeing of mice provided various enrichments. In this study, we used female ICR mice to evaluate enrichment strategies that encouraged natural behaviors including foraging, exercise, sheltering, and socialization. After 3 mo of exposure to the assigned enrichment strategy, wellbeing was assessed by evaluating behavioral and physiologic differences between groups. The results suggested that the use of red-tinted igloos may decrease markers of mouse wellbeing. However, none of the selected strategies yielded measures of wellbeing indicating improvement as compared to individually housed mice with no enrichment (negative control). Furthermore, measures were not significantly different between paired mice and individually housed mice with no enrichment.

Comparison of 2 transabdominal ultrasound image guidance techniques for prostate and prostatic fossa radiation therapy.

PURPOSE: Our clinic is a long-term user of a first-generation transabdominal (TA) biplanar (2.5-dimensional [2.5D]) ultrasound image guidance (USIG) system for prostate cancer treatments. We are also an early adopter and development partner for a new, second-generation, fully 3D USIG system that allows for volumetric TA localization of the prostate. This new system has been evaluated at our institution by direct comparison with the previously established first-generation TA method for prostate alignment. METHODS AND MATERIALS: We compared the 2 TA-USIG methods on the same subjects and same treatment sessions. A total of 1428 fractions delivered to 41 treated patients (16 intact prostate, 25 fossa) were analyzed regarding the agreement of alignments between the 2 US positioning systems. Patients were first aligned to tattoos using treatment room lasers. TA-USIG using the 3D system was then performed to align contours derived during the computed tomography simulation process to their corresponding daily US-visualized structures. The US-3D system image guidance shifts were performed and recorded as the "initial" shifts. A 2.5D system alignment was then immediately performed using the same computed tomography derived reference contours and the indicated shifts, relative to the 3D system, were recorded as the difference between the 2 alignment methods. RESULTS: The average difference between the 2 TA-USIG alignments for all patients was 0.4 ± 0.7 mm, 0.7 ± 0.9 mm, and 0.5 ± 0.9 mm in the left-right, anteroposterior, and superoinferior directions, respectively. No significant difference in system agreement between intact prostate versus fossa patients was observed. CONCLUSION: Our comparison of an established 2.5D USIG method with a newer, fully 3D approach for prostate alignment of 41 different patients (1428 fractions) shows excellent agreement with each other, despite the nontrivial difference in imaging approaches. This shows that the 2 specific USIG approaches yield results that are consistent with each other, and that the USIG modality yields consistent results within the modality.

Complications of elastase-induced arterial saccular aneurysm in rabbits: case reports and literature review.

Endoluminal infusion and incubation of elastase with or without collagenase into the rabbit common carotid artery is an established model of arterial saccular aneurysm. The model mimics naturally occurring human cerebral aneurysms in many ways, including histologic and morphologic characteristics, hemodynamic pressures, and shear stresses. However, complications have been associated with the model. Here, we report 2 complications: 1) the first known case of iatrogenic laryngeal hemiplegia in a rabbit; and 2) histopathologically confirmed iatrogenic hippocampal and cerebellar infarcts (stroke). Finally, we present and review data from current literature on the morbidity and mortality associated with this model.

Midazolam enhances the analgesic properties of dexmedetomidine in the rat.

OBJECTIVE: To investigate the analgesic properties of different dose combinations of midazolam and dexmedetomidine administered intraperitoneally (IP) in the rat. STUDY DESIGN: Prospective experimental trial. ANIMALS: Seventy adult male Sprague Dawley rats weighing 250-300 g. METHODS: Dexmedetomidine (D) 0.03, 0.06, 0.09, 0.12, 0.15, 0.18, 0.21 mg kg(-1) and midazolam (M) 5, 10, 25, 50 mg kg(-1) were administered IP, alone then in combinations ranging from 0.03 D:5 M to 0.18 D:30 M mg kg(-1). Analgesia was evaluated using the tail-flick test at time 0 (before injection), 15, 30, 45, 60 and 75 minutes. RESULTS: Midazolam at all doses administered (5-50 mg kg(-1)) did not significantly change tail-flick latencies from baseline values whereas D showed clear dose-dependent increases in tail-flick latency for doses administered in the range of 0.03-0.18 mg kg(-1). Tail-flick latencies in rats administered D+M combinations were significantly greater than D alone (p<0.05). CONCLUSIONS: A dose-related analgesic effect was demonstrated for D in the rat, which was enhanced by co-administration of M. CLINICAL RELEVANCE: The combination of D+M administered IP to rats at doses of 0.12:20 and 0.09:15 mg kg(-1) was shown to be a good combination to provide sedation/analgesia with a duration of action greater than 60 minutes. The onset of sedation was rapid (1-3 minutes), and onset of profound analgesia was reached within 5-10 minutes.

The functional genome of CA1 and CA3 neurons under native conditions and in response to ischemia.

BACKGROUND: The different physiological repertoire of CA3 and CA1 neurons in the hippocampus, as well as their differing behaviour after noxious stimuli are ultimately based upon differences in the expressed genome. We have compared CA3 and CA1 gene expression in the uninjured brain, and after cerebral ischemia using laser microdissection (LMD), RNA amplification, and array hybridization. RESULTS: Profiling in CA1 vs. CA3 under normoxic conditions detected more than 1000 differentially expressed genes that belong to different, physiologically relevant gene ontology groups in both cell types. The comparison of each region under normoxic and ischemic conditions revealed more than 5000 ischemia-regulated genes for each individual cell type. Surprisingly, there was a high co-regulation in both regions. In the ischemic state, only about 100 genes were found to be differentially expressed in CA3 and CA1. The majority of these genes were also different in the native state. A minority of interesting genes (e.g. inhibinbetaA) displayed divergent expression preference under native and ischemic conditions with partially opposing directions of regulation in both cell types. CONCLUSION: The differences found in two morphologically very similar cell types situated next to each other in the CNS are large providing a rational basis for physiological differences. Unexpectedly, the genomic response to ischemia is highly similar in these two neuron types, leading to a substantial attenuation of functional genomic differences in these two cell types. Also, the majority of changes that exist in the ischemic state are not generated de novo by the ischemic stimulus, but are preexistant from the genomic repertoire in the native situation. This unexpected influence of a strong noxious stimulus on cell-specific gene expression differences can be explained by the activation of a cell-type independent conserved gene-expression program. Our data generate both novel insights into the relation of the quiescent and stimulus-induced transcriptome in different cells, and provide a large dataset to the research community, both for mapping purposes, as well as for physiological and pathophysiological research.

Global transcriptome analysis of genetically identified neurons in the adult cortex.

The enormous cellular complexity of the brain is a major obstacle for gene expression profiling of neurological disease models, because physiologically relevant changes of transcription in a specific neuronal subset are likely to be lost in the presence of other neurons and glia. We solved this problem in transgenic mice by labeling genetically defined cells with a nuclear variant of GFP. When combined with laser-directed microdissection, intact RNA from unfixed, freeze-dried sections can be isolated, which is a prerequisite for high-quality global transcriptome analysis. Here, we compared gene expression profiles between pyramidal motor neurons and pyramidal somatosensory neurons captured from layer V of the adult neocortex. One striking feature of motor neurons is the elevated expression of ribosomal genes and genes involved in ATP synthesis. This suggests a molecular adaptation of the upper motor neurons to longer axonal projections and higher electrical activity. These molecular signatures were not detected when cortical layers and microareas were analyzed in toto. Additionally, we used microarrays to determine the global mRNA expression profiles of microdissected Purkinje cells and cellularly complex cerebellar cortex microregions. In summary, our analysis shows that cellularly complex targets lead to averaged gene expression profiles that lack substantial amounts of cell type-specific information. Thus, cell type-restricted sampling strategies are mandatory in the CNS. The combined use of a genetic label with laser-microdissection offers an unbiased approach to map patterns of gene expression onto practically any cell type of the brain.