A Simple, Inexpensive, and Effective Light- Carrying Laryngoscopic Blade for Orotracheal Intubation of Rats

The research paradigm of using large laboratory animals, in which oroendotracheal intubations are relatively easy, is shifting toward the use of small animals, such as rodents, in which oropharyngeal access is limited, the arytenoid cartilage cycles are faster, and the glottis is much smaller. The considerable growth recently seen in preclinical imaging studies is accompanied by an increased number of rats and mice requiring in vivo intubation for airway management. Tracheal access is important for ventilation, administration of inhaled anesthetics, instillation of drugs or imaging agents, and maintenance of airway patency to reduce mortality during and after operations. I fashioned a light-carrying laryngoscopic blade (laryngoscope) from readily available acrylic-polymethyl methacrylate tubing and used it to perform rapid, effective tracheal intubation in rats. The laryngoscope design and intubation techniques are presented

The Effect of ACE Inhibition on the Pulmonary Vasculature in Combined Models of Chronic Hypoxia and Pulmonary Arterial Banding in Sprague Dawley Rats

Microfocal CT was used to image the pulmonary arterial (PA) tree in rodent models of pulmonary hypertension (PH). CT images were used to measure the arterial tree diameter along the main arterial trunk at several hydrostatic intravascular pressures and calculate distensibility. High-resolution planar angiographic imaging was also used to examine distal PA microstructure. Data on pulmonary artery tree morphology improves our understanding of vascular remodeling and response to treatments. Angiotensin II (ATII) has been identified as a mediator of vasoconstriction and proliferative mitotic function. ATII has been shown to promote vascular smooth muscle cell hypertrophy and hyperplasia as well as stimulate synthesis of extracellular matrix proteins. Available ATII is targeted through angiotensin converting enzyme inhibitors (ACEIs), a method that has been used in animal models of PH to attenuate vascular remodeling and decrease pulmonary vascular resistance. In this study, we used rat models of chronic hypoxia to induce PH combined with partial left pulmonary artery occlusion (arterial banding, PLPAO) to evaluate effects of the ACEI, captopril, on pulmonary vascular hemodynamic and morphology. Male Sprague Dawley rats were placed in hypoxia (FiO2 0.1), with one group having underwent PLPAO three days prior to the chronic hypoxia. After the twenty-first day of hypoxia exposure, treatment was started with captopril (20 mg/kg/day) for an additional twenty-one days. At the endpoint, lungs were excised and isolated to examine: pulmonary vascular resistance, ACE activity, pulmonary vessel morphology and biomechanics. Hematocrit and RV/LV+septum ratio was also measured. CT planar images showed less vessel dropout in rats treated with captopril versus the non-treatment lungs. Distensibility data shows no change in rats treated with captopril in both chronic hypoxia (CH) and CH with PLPAO (CH+PLPAO) models. Hemodynamic measurements also show no change in the pulmonary vascular resistance with captopril treatment in both CH and CH+PLPAO

Semiautomated Skeletonization of the Pulmonary Arterial Tree in Micro-CT Images

We present a simple and robust approach that utilizes planar images at different angular rotations combined with unfiltered back-projection to locate the central axes of the pulmonary arterial tree. Three-dimensional points are selected interactively by the user. The computer calculates a sub- volume unfiltered back-projection orthogonal to the vector connecting the two points and centered on the first point. Because more x-rays are absorbed at the thickest portion of the vessel, in the unfiltered back-projection, the darkest pixel is assumed to be the center of the vessel. The computer replaces this point with the newly computer-calculated point. A second back-projection is calculated around the original point orthogonal to a vector connecting the newly-calculated first point and user-determined second point. The darkest pixel within the reconstruction is determined. The computer then replaces the second point with the XYZ coordinates of the darkest pixel within this second reconstruction. Following a vector based on a moving average of previously determined 3- dimensional points along the vessel\u27s axis, the computer continues this skeletonization process until stopped by the user. The computer estimates the vessel diameter along the set of previously determined points using a method similar to the full width-half max algorithm. On all subsequent vessels, the process works the same way except that at each point, distances between the current point and all previously determined points along different vessels are determined. If the difference is less than the previously estimated diameter, the vessels are assumed to branch. This user/computer interaction continues until the vascular tree has been skeletonized

Post-Acquisition Small-Animal Respiratory Gated Imaging Using Micro Cone-Beam CT

On many occasions, it is desirable to image lungs in vivo to perform a pulmonary physiology study. Since the lungs are moving, gating with respect to the ventilatory phase has to be performed in order to minimize motion artifacts. Gating can be done in real time, similar to cardiac imaging in clinical applications, however, there are technical problems that have lead us to investigate different approaches. The problems include breath-to-breath inconsistencies in tidal volume, which makes the precise detection of ventilatory phase difficult, and the relatively high ventilation rates seen in small animals (rats and mice have ventilation rates in the range of a hundred cycles per minute), which challenges the capture rate of many imaging systems (this is particularly true of our system which utilizes cone-beam geometry and a 2 dimensional detector). Instead of pre-capture ventilation gating we implemented a method of post-acquisition gating. We acquire a sequence of projections images at 30 frames per second for each of 360 viewing angles. During each capture sequence the rat undergoes multiple ventilation cycles. Using the sequence of projection images, an automated region of interest algorithm, based on integrated grayscale intensity, tracts the ventilatory phase of the lungs. In the processing of an image sequence, multiple projection images are identified at a particular phase and averaged to improve the signal-to-ratio. The resulting averaged projection images are input to a Feldkamp cone-beam algorithm reconstruction algorithm in order to obtain isotropic image volumes. Minimal motion artifact data sets improve qualitative and quantitative analysis techniques useful in physiologic studies of pulmonary structure and function

Quasi-Exact Helical Cone Beam Reconstruction for Micro CT

A cone beam micro-CT system is set up to collect truncated helical cone beam data. This system includes a micro-focal X-ray source, a precision computer-controlled X-Y-Z-theta stage, and an image-intensifier coupled to a large format CCD detector. The helical scanning mode is implemented by rotating and translating the stage while keeping X-ray source and detector stationary. A chunk of bone and a mouse leg are scanned and quasi-exact reconstruction is performed using the approach proposed in J. Hu et al. (2001). This approach introduced the original idea of accessory paths with upper and lower virtual detectors having infinite axial extent. It has a filtered backprojection structure which is desirable in practice and possesses the advantages of being simple to implement and computationally efficient compared to other quasi-exact helical cone beam algorithms for the long object problem

Measuring the effect of airway pressure on pulmonary arterial diameter in the intact rat lung

To study the relationship between transpulomnary pressure (Ptp), intravascular pressure (Pv), and the pulmonary arterial tree structure, morphometric measurements of pulmonary arterial trees were made in intact lungs from Sprague-Dawley rats. Using cone beam micro-CT and techniques we developed for imaging small animal lungs, volumetric CT data were acquired for Ptp from 0 - 12 mmHg and Pv from 5 - 30 mmHg. The diameter, D (measured range approximately 0.08-2.0 mm), vs. pressure, P, relation can be described by D(P) = D(0)(1+ α P), where α is a distensibility coefficient. Unlike studies performed in larger animals, where changes in either Ptp or Pv had nearly identical effect on vessel distensibility, we found that there is only a small dependence of arterial diameter on Ptp in the rat. For example, using the above relation where P=Ptp and Pv is held constant at 12mmHg, alpha = 0.55±0.42(SE) %/mmHg, compared with when P=Pv and Ptp is held at 12mmHg, alpha = 2.59±0.17(SE) %/mmHg

Changes in Pulmonary Arterial Wall Mechanical Properties and Lumenal Architecture with Induced Vascular Remodeling

To explore and quantify pulmonary arterial remodeling we used various methods including micro-CT, high-resolution 3-dimensional x-ray imaging, to examine the structure and function of intact pulmonary vessels in isolated rat lungs. The rat is commonly used as an animal model for studies of pulmonary hypertension (PH) and the accompanying vascular remodeling, where vascular remodeling has been defined primarily by changes in the vessel wall composition in response to hypertension inducing stimuli such as chronic hypoxic exposure (CHE) or monocrotaline (MCT) injection. Little information has been provided as to how such changes affect the vessel wall mechanical properties or the lumenal architecture of the pulmonary arterial system that actually account for the hemodynamic consequences of the remodeling. In addition, although the link between primary forms of pulmonary hypertension and inherited genetics is well established, the role that genetic coding plays in hemodynamics and vascular remodeling is not. Therefore, we are utilizing Fawn-Hooded (FH), Sprague-Dawley (SD) and Brown Norway (BN)rat strains along with unique imaging methods to parameterize both vessel distensibility and lumenal morphometry using a principal pulmonary arterial pathway analysis based on self-consistency. We have found for the hypoxia model, in addition to decreased body weight, increased hematocrit, increased right ventricular hypertrophy, the distensibility of the pulmonary arteries is shown to decrease significantly in the presence of remodeling