Richard P. Kennan

Neurological and behavioral abnormalities, ventricular dilatation, altered cellular functions, inflammation, and neuronal injury in brains of mice due to common, persistent, parasitic infection

BackgroundWorldwide, approximately two billion people are chronically infected with Toxoplasma gondii with largely unknown consequences.MethodsTo better understand long-term effects and pathogenesis of this common, persistent brain infection, mice were infected at a time in human years equivalent to early to mid adulthood and studied 5–12 months later. Appearance, behavior, neurologic function and brain MRIs were studied. Additional analyses of pathogenesis included: correlation of brain weight and neurologic findings; histopathology focusing on brain regions; full genome microarrays; immunohistochemistry characterizing inflammatory cells; determination of presence of tachyzoites and bradyzoites; electron microscopy; and study of markers of inflammation in serum. Histopathology in genetically resistant mice and cytokine and NRAMP knockout mice, effects of inoculation of isolated parasites, and treatment with sulfadiazine or αPD1 ligand were studied.ResultsTwelve months after infection, a time equivalent to middle to early elderly ages, mice had behavioral and neurological deficits, and brain MRIs showed mild to moderate ventricular dilatation. Lower brain weight correlated with greater magnitude of neurologic abnormalities and inflammation. Full genome microarrays of brains reflected inflammation causing neuronal damage (Gfap), effects on host cell protein processing (ubiquitin ligase), synapse remodeling (Complement 1q), and also increased expression of PD-1L (a ligand that allows persistent LCMV brain infection) and CD 36 (a fatty acid translocase and oxidized LDL receptor that mediates innate immune response to beta amyloid which is associated with pro-inflammation in Alzheimers disease). Immunostaining detected no inflammation around intra-neuronal cysts, practically no free tachyzoites, and only rare bradyzoites. Nonetheless, there were perivascular, leptomeningeal inflammatory cells, particularly contiguous to the aqueduct of Sylvius and hippocampus, CD4+ and CD8+ T cells, and activated microglia in perivascular areas and brain parenchyma. Genetically resistant, chronically infected mice had substantially less inflammation.ConclusionIn outbred mice, chronic, adult acquired T. gondii infection causes neurologic and behavioral abnormalities secondary to inflammation and loss of brain parenchyma. Perivascular inflammation is prominent particularly contiguous to the aqueduct of Sylvius and hippocampus. Even resistant mice have perivascular inflammation. This mouse model of chronic T. gondii infection raises questions of whether persistence of this parasite in brain can cause inflammation or neurodegeneration in genetically susceptible hosts.

Effects of susceptibility variations on NMR measurements of diffusion

Abstract A pulsed-gradient spin-echo method has been used to measure the water self-diffusion coefficient in various media containing susceptibility variations arising from the presence of superparamagnetic iron oxide particles. These include aqueous suspensions, polyacrylamide gels, and rat livers. The results for each medium indicate that, contrary to intuition, the apparent diffusion coefficient decreases as the iron particle concentration increases. A simple theoretical model has been developed to interpret the data. This model relates the changes in apparent diffusion to the variance of the internal gradient distribution generated by the iron particles. A computer simulation of nuclei undergoing a random walk in an inhomogeneous medium was performed and the results provide qualitative confirmation of the experimental observations. The relevance of these phenomena to measurements of molecular diffusion in heterogeneous systems and their use to estimate internal gradient strengths are discussed.

Non-invasive assessment of language lateralization by transcranial near infrared optical topography and functional MRI.

Near infrared optical topography (OT) is the simultaneous acquisition of hemoglobin absorption from an array of optical fibers on the scalp to construct maps of cortical activity. We demonstrate that OT can be used to determine lateralization of prefrontal areas to a language task that has been validated by functional MRI (fMRI). Studies were performed on six subjects using a visually presented language task. Laterality was quantified by the relative number of activated pixels in each hemisphere for fMRI, and the total hemoglobin responses in each hemisphere for OT. All subjects showed varying degrees of left hemisphere language dominance and the mean laterality indices for subjects who underwent both OT and fMRI were in good agreement. These studies demonstrate that OT gives predictions of hemispheric dominance that are consistent with fMRI. Due to the ease of use and portable nature of OT, it is anticipated that optical topography will be valuable tool for neurological examinations of cognitive function. Hum. Brain Mapping 16:183–189, 2002.

A model for susceptibility artefacts from respiration in functional echo-planar magnetic resonance imaging

Respiration causes variations in the signals acquired during magnetic resonance imaging (MRI) and therefore is a significant source of noise in functional brain imaging. A primary component of respiratory noise may arise from variations of bulk susceptibility or air volume in the chest. Here we investigate the nature of the image artefacts that can be caused by such changes. We develop a simple model which attempts to mimic the effects of variations in susceptibility and volume during respiration. Theoretical calculations, computer simulations and imaging experiments with this model show that small variations in susceptibility within the thorax from alterations in the paramagnetism of cavity gas may lead to a shift of the image on the order of 0.1 pixels as well as a shading of the intensity by +/-1%. These effects are observed to be predominant in the phase-encoding direction. They may lead to the production of spurious activations in functional MRI and are likely to be of more importance at higher field strengths.

High-Resolution CMRO2 Mapping in Rat Cortex: A Multiparametric Approach to Calibration of BOLD Image Contrast at 7 Tesla:

The blood oxygenation level-dependent (BOLD) functional magnetic resonance imaging (fMRI) method, which is sensitive to vascular paramagnetic deoxyhemoglobin, is dependent on regional values of cerebral metabolic rate of oxygen utilization (CMRO2), blood flow (CBF), and volume (CBV). Induced changes in deoxyhemoglobin function as an endogenous contrast agent, which in turn affects the transverse relaxation rates of tissue water that can be measured by gradient-echo and spin-echo sequences in BOLD fMRI. The purpose here was to define the quantitative relation between BOLD signal change and underlying physiologic parameters. To this end, magnetic resonance imaging and spectroscopy methods were used to measure CBF, CMRO2, CBV, and relaxation rates (with gradient-echo and spin-echo sequences) at 7 Tesla in rat sensorimotor cortex, where cerebral activity was altered pharmacologically within the autoregulatory range. The changes in tissue transverse relaxation rates were negatively and linearly correlated with changes in CBF, CMRO2, and CBV. The multiparametric measurements revealed that CBF and CMRO2 are the dominant physiologic parameters that modulate the BOLD fMRI signal, where the ratios of (ΔCMRO2/CMRO2)/(ΔCBF/CBF) and (ΔCBV/CBV)/(ΔCBF/CBF) were 0.86 ± 0.02 and 0.03 ± 0.02, respectively. The calibrated BOLD signals (spatial resolution of 48 μL) from gradient-echo and spin-echo sequences were used to predict changes in CMRO2 using measured changes in CBF, CBV, and transverse relaxation rates. The excellent agreement between measured and predicted values for changes in CMRO2 provides experimental support of the current theory of the BOLD phenomenon. In gradient-echo sequences, BOLD contrast is affected by reversible processes such as static inhomogeneities and slow diffusion, whereas in spin-echo sequences these effects are refocused and are mainly altered by extravascular spin diffusion. This study provides steps by which multiparametric MRI measurements can be used to obtain high-spatial resolution CMRO2 maps.

Dependence of Oxygen Delivery on Blood Flow in Rat Brain: A 7 Tesla Nuclear Magnetic Resonance Study

Magnetic resonance imaging (MRI) and spectroscopy (MRS) were used at a magnetic field strength of 7 T to measure CBF and CMRO2 in the sensorimotor cortex of mature rats at different levels of cortical activity. In rats maintained on morphine anesthesia, transitions to lower activity and higher activity states were produced by administration of pentobarbital and nicotine, respectively. Under basal conditions of morphine sulfate anesthesia, CBF was 0.75 ± 0.09 mL · g−1 · min−1 and CMRO2 was 3.15 ± 0.18 μmol · g−1 · min−1. Administration of sodium pentobarbital reduced CBF and CMRO2 by 66% ± 16% and 61% ± 6%, respectively (i.e., “deactivation”). In contrast, administration of nicotine hydrogen tartrate increased CBF and CMRO2 by 41% ± 5% and 30% ± 3%, respectively (i.e., “activation”). The resting values of CBF and CMRO2 for α-chloralose anesthetized rats were 0.40 ± 0.09 mL · g−1 · min−1 and 1.51 ± 0.06 μmol · g−1 · min−1, respectively. Upon forepaw stimulation, CBF and CMRO2 were focally increased by 34% ± 10% and 26% ± 12%, respectively, above the resting nonanesthetized values (i.e., “activation”). Incremental changes in CBF and CMRO2, when expressed as a percentage change for “deactivation” and “activation” from the respective control conditions, were linear (R2 = 0.997) over the entire range examined with the global and local perturbations. This tight correlation for cerebral oxygen delivery in vivo is supported by a recent model where the consequence of a changing effective diffusivity of the capillary bed for oxygen, D, has been hypothetically shown to be linked to alterations in CMRO2 and CBF. This assumed functional characteristic of the capillary bed can be theoretically assessed by the ratio of fractional changes in D with respect to changes in CBF, signified by Ω. A value 0.81 ± 0.23 was calculated for Ω with the in vivo data presented here, which in turn corresponds to a supposition that the effective oxygen diffusivity of the capillary bed is not constant but presumably varies to meet local requirements in oxygen demand in a similar manner with both “deactivation” and “activation.”

Assessment of heating effects in skin during continuous wave near infrared spectroscopy

Near infrared spectroscopy is an increasingly important tool for the investigation of human brain function, however, to date there have been few systematic evaluations of accompanying thermal effects due to absorption. We have measured the spatial distribution of temperature changes during near infrared irradiation (789 nm) as a function of laser power, in both excised tissue (chicken meat and skin) and in the forearm of an awake human volunteer. Light was applied using a 1 mm optical fiber which is characteristic of the topographic system. The temperature of excised chicken tissue increased linearly with power level as 0.097 and 0.042 degrees C/mW at depths of 0 and 1 mm, respectively. Human forearm studies yielded temperature changes of 0.101, 0.038, and 0.030 degrees C/mW at depths of 0.5, 1.0, and 1.5 mm, respectively. Due to direct irradiation of the thermocouple all measurements represent the maximum temperature increase from the laser. In all cases the estimated heating effects from continuous wave optical topography systems were small and well below levels which would endanger tissue cells. The close similarity between ex vivo and in vivo measurements suggests negligible contributions from blood flow in the skin which was further supported by measurements during cuff ischemia. Heating effects decreased sharply with both depth and lateral position; thus, for optode spacings greater than a few millimeters, fibers can be treated independently. Finite element analysis confirms that the experimental results are consistent with a simple heat conduction model.

Quantitative imaging of magnetization transfer using multiple selective pulses.

New spectroscopic and imaging methods have been developed for quantitatively measuring magnetization transfer (MT). These methods use trains of radiofrequency (rf) pulses with pulse separations much longer than 1/kmf and pulse durations much shorter than 1/kmf, where kmf is the rate of MT from the immobile (macromolecular) protons to the mobile (free water) protons. Signal sensitivity to MT occurs when these pulses affect the mobile and immobile proton pools to different degrees. The signal from water may be quantitatively related to the macromolecular content of the sample using theory. The method has been used to make quantitative measurements of macromolecular content in cross‐linked bovine serum albumin and employed in conjunction with echoplanar imaging to produce maps of the spatial distribution of the macromolecular content. Magn Reson Med 41:1065–1072, 1999.

A general model of microcirculatory blood flow effects in gradient sensitized MRI

A general expression is derived for the NMR signal from a fluid undergoing random directional flow such as encountered within the microcirculation. The dependence of the echo amplitude on flow velocity, sample morphology, and experimental parameters are described in terms of a temporal velocity autocorrelation function. The width of the correlation function determines whether the flow can properly be described as diffusive. Comparison is made between the velocity autocorrelation method outlined here and the IVIM model for tissue perfusion. Conditions for the validity of the latter approach for extracting physiologic information from apparent diffusion measurements are discussed. The approach outlined leads to a more robust measure of microcirculatory blood velocity from NMR measurements.

Decreased cerebral perfusion correlates with increased BOLD hyperoxia response in transgenic mouse models of sickle cell disease

Neurological complications such as stroke are known consequences of sickle cell disease (SCD). In order to improve methods for the evaluation of stroke risk in SCD, MRI was used to evaluate cerebrovascular function in transgenic mouse models of human SCD. It is hypothesized that oxygen‐sensitive imaging in the brain will reveal areas of excess deoxygenation that are either at risk of or the result of vaso‐occlusion. Arterial spin labeling (ASL) perfusion was performed in order to correlate BOLD results with microvascular cerebral blood flow. Upon comparison with control animals, there was a relative increase in BOLD hyperoxia response of 42–67% (P < 0.001) in the transgenic mice while cerebral blood flow during normoxia was reduced by 30–40% (P < 0.02). Hyperoxia caused cerebral blood flow to decrease in control mice, whereas blood flow increased in the sickle transgenic mice. These results indicate impairment in brain autoregulation in the sickle cell transgenic mice leading to increased cerebral deoxyhemoglobin. Increased deoxyhemoglobin coupled with reduced perfusion may further increase the risk of vaso‐occlusion and stroke. This may reflect polymer reduction or reduced cell adhesion during hyperoxia. The MRI protocol is noninvasive and thus directly applicable to a clinical population. Magn Reson Med 51:525–532, 2004.