Gilbert R. Hillman

Serotonin2C receptor localization in GABA neurons of the rat medial prefrontal cortex: implications for understanding the neurobiology of addiction.

Serotonin (5-HT) action via the 5-HT(2C) receptor (5-HT(2C)R) provides an important modulatory influence over neurons of the prefrontal cortex (PFC), which is critically involved in disorders of executive function including substance use disorders. In the present study, we investigated the distribution of the 5-HT(2C)R in the rat prelimbic prefrontal cortex (PrL), a subregion of the medial prefrontal cortex (mPFC), using a polyclonal antibody raised against the 5-HT(2C)R. The expression of 5-HT(2C)R immunoreactivity (IR) was highest in the deep layers (layers V/VI) of the mPFC. The 5-HT(2C)R-IR was typically most intense at the periphery of cell bodies and the initial segment of cell processes. Approximately 50% of the 5-HT(2C)R-IR detected was found in glutamate decarboxylase, isoform 67 (GAD 67)-positive neurons. Of the subtypes of GABA interneurons identified by expression of several calcium-binding proteins, a significantly higher percentage of neurons expressing IR for parvalbumin also expressed 5-HT(2C)R-IR than did the percentage of neurons expressing calbindin-IR or calretinin-IR that also expressed 5-HT(2C)R-IR. Since parvalbumin is located in basket and chandelier GABA interneurons which project to cell body and initial axon segments of pyramidal cells, respectively, these results raise the possibility that the 5-HT(2C)R in the mPFC acts via the parvalbumin-positive GABAergic interneurons to regulate the output of pyramidal cells in the rat mPFC.

Perfusion Deficit Parallels Exacerbation of Cerebral Ischemia/Reperfusion Injury in Hyperglycemic Rats

Magnetic resonance imaging (MRI) techniques were used to determine the effect of preexisting hyperglycemia on the extent of cerebral ischemia/reperfusion injury and the level of cerebral perfusion. Middle cerebral artery occlusion (MCAO) was induced by a suture insertion technique. Forty one rats were divided into hyperglycemic and normoglycemic groups with either 4 hours of continuous MCAO or 2 hours of MCAO followed by 2 hours of reperfusion. Diffusion-weighted imaging (DWI) was performed at 4 hours after MCAO to quantify the degree of injury in 6 brain regions. Relative cerebral blood flow (CBF) and cerebral blood volume (CBV) were estimated using gradient echo (GE) bolus tracking and steady-state spin echo (SE) imaging techniques, respectively. Brain injury correlated with the perfusion level measured in both SE CBV and dynamic GE CBF images. In the temporary MCAO model, mean lesion size in DWI was 118% larger and hemispheric CBV was reduced by 37% in hyperglycemic compared with normoglycemic rats. Hyperglycemia did not significantly exacerbate brain injury or CBV deficit in permanent MCAO models. We conclude that preexisting hyperglycemia increases acute postischemic MRI-measurable brain cellular injury in proportion to an associated increased microvascular ischemia.

CHRONIC PHENCYCLIDINE INCREASES NMDA RECEPTOR NR1 SUBUNIT MRNA IN RAT FOREBRAIN

The present study was designed to determine whether the sensitization of locomotor activity that results from chronic phencyclidine (PCP) administration is associated with altered NMDA receptor function or mRNA in rat brain. Female Sprague‐Dawley rats were administered PCP (20 mg/kg, i.p.) once daily for 5 days. After withdrawal for 72 hr, challenge with 3.2 mg/kg PCP (i.p.) revealed a significant sensitization to the locomotor activating effect of PCP. In situ hybridization analysis with an oligonucleotide probe complementary to the mRNA encoding the NR1 subunit of the NMDA receptor demonstrated that chronic PCP treatment resulted in a marked increase in NR1 subunit mRNA in the forebrain. Quantitative image analysis revealed a significant increase in the labeling of NR1 mRNA in the olfactory tubercle, piriform cortex, frontal cortex, and anterior striatum. However, no significant difference between PCP and saline‐treated rats was found in the hippocampus or cerebellum. In a parallel study, possible functional alterations in the NMDA receptor were assessed by measuring NMDA‐stimulated release of [3H]DA from slices of the olfactory tubercle and piriform cortex. NMDA‐stimulated release was not affected by chronic PCP treatment, but the inhibition of this release by PCP, 7‐chlorokynurenic acid (7‐CK), and DL‐2‐amino‐5‐phosphovaleric acid (AP‐5) was significantly diminished by chronic PCP. This suggests that the behavioral plasticity associated with chronic PCP may be related to an altered subunit stoichiometry of NMDA receptors in selective forebrain regions. J. Neurosci. Res. 55:762–769, 1999. 

Measurement of carpal bone geometry by computer analysis of three-dimensional CT images

The aim of this project was noninvasively to analyze and quantitate the geometry, load transfer characteristics, and spatial relationships of the carpal bones by using a new three-dimensional CT scan reconstruction technique. The determination of mechanical parameters such as distances between centroids and between bone surfaces, carpal alignment, volumes, surface areas, and contact areas can provide the basis for comparison between normal wrists and wrists with a variety of progressive instability patterns, types of fracture, pathologic and posttraumatic states, and different simulated surgical procedures. This new technology has demonstrated a volumetric accuracy of 94% and a linear accuracy of 97%. Simultaneous analysis of all articulating surfaces of multiple joints can be performed in cadavers and in patients because of the noninvasive nature of the imaging reconstruction technique. This new research offers much more information than has previously been available. It also promises direct application to the clinical setting and eliminates several limitations and questions that were inescapable with previous technology.

Augmentation of locomotor activity by chronic phencyclidine is associated with an increase in striatal NMDA receptor function and an upregulation of the NR1 receptor subunit

Phencyclidine (PCP) is a drug of abuse that produces schizophrenia‐like symptoms in humans and increases locomotor activity and stereotypic behavior in rodents. PCP‐induced alteration in rat locomotor activity is thought to be mediated by an inhibition of N‐methyl‐D‐aspartate (NMDA) receptors in the striatum and other brain regions. In this study, rats treated chronically with PCP (20 mg/kg once per day for 5 days) showed a marked increase in locomotor activity following a PCP challenge (3.2 mg/kg) administered after either 3 or 8 days of withdrawal. In biochemical assays, the release of striatal [14C]GABA by NMDA was enhanced by about 77% by chronic PCP treatment, whereas [3H]ACh release was increased by about 31% in tissue from PCP‐treated rats. Even though binding experiments with 1‐[1‐(2‐thiehyl)cyclohexyl]piperidyl‐3,4 3H(N) ([3H]TCP) showed no alteration in the Kd or Bmax in whole striatum, quantitative immunocytochemical experiments found an upregulation in the NR1 subunit in the cell bodies and neuropil of cortical and striatal regions of the forebrain following chronic PCP treatment. An increase in the size of NR1‐immunoreactive cells in the forebrain was also observed following chronic PCP treatment. Together, these data may help in understanding the mechanisms underlying the adaptive response to chronic reduction in glutamatergic NMDA transmission that has been postulated to be involved in the etiology of schizophrenia. Synapse 31:229–239, 1999.

An affine invariant curve matching method for photo-identification of marine mammals

Individual identification of marine mammals is of interest to marine biologists. This paper aims at the recognition of the edges associated with marine mammals whose pictures are taken under affine transformations. The introduced affine curve matching method uses the area of mismatch between a query and a database curve. This area is obtained by optimally aligning the curves based on the minimum affine distance involving their distinguishing points. The method is applied to databases of sea lions, gray whales, and dolphins, and its performance is compared with two other affine curve matching methods. The results show that the introduced curve matching approach outperforms the other approaches and thus further reduces the search time for identifying an individual. The developed method is of general purpose as it can be used for other affine curve matching applications.

A string matching computer-assisted system for dolphin photoidentification.

AbstractThis paper presents a syntactic/semantic string representation scheme as well as a string matching method as part of a computer-assisted system to identify dolphins from photographs of their dorsal fins. A low-level string representation is constructed from the curvature function of a dolphins fin trailing edge, consisting of positive and negative curvature primitives. A high-level string representation is then built over the low-level string via merging appropriate groupings of primitives in order to have a less sensitive representation to curvature fluctuations or noise. A family of syntactic/semantic distance measures between two strings is introduced. A composite distance measure is then defined and used as a dissimilarity measure for database search, highlighting both the syntax (structure or sequence) and semantic (attribute or feature) differences. The syntax consists of an ordered sequence of significant protrusions and intrusions on the edge, while the semantics consist of seven attributes extracted from the edge and its curvature function. The matching results are reported for a database of 624 images corresponding to 164 individual dolphins. The identification results indicate that the developed string matching method performs better than the previous matching methods including dorsal ratio, curvature, and curve matching. The developed computer-assisted system can help marine mammalogists in their identification of dolphins, since it allows them to examine only a handful of candidate images instead of the currently used manual searching of the entire database.

Frontal Lobe Changes after Severe Diffuse Closed Head Injury in Children

IN VIEW OF the pathophysiology and biomechanics of severe closed head injury (CHI) in children, we postulated that the frontal lobes sustain diffuse injury, even in the absence of focal brain lesions detected by magnetic resonance imaging (MRI). This study quantitated the morphological effects of CHI on the frontal lobes in children who sustained head trauma of varying severity. The MRI findings of 14 children who had sustained severe CHls (Glasgow Coma Scale score of ≤8) were compared with the findings in a matched group of 14 children having sustained mild head injuries (Glasgow Coma Scale score of 13-15). The patients ranged in age from 5 to 15 years at the time of their MRls, which were acquired at least 3 months postinjury. MRI findings revealed no focal areas of abnormal signal in the frontal lobes. Volumetric analysis disclosed that the total prefrontal cerebrospinal fluid increased and the gray matter volume decreased in the patients with severe CHI, relative to the mildly injured comparison group. Gray matter volume was also reduced in the orbitofrontal and dorsolateral regions of the brains of children with severe CHI, relative to the children who sustained mild head trauma. These volumetric findings indicate that prefrontal tissue loss occurs after severe CHI in children, even in the absence of focal brain lesions in this area. Nearly two-thirds of the children who sustained severe CHls were moderately disabled after an average postinjury interval of 3 years or more, whereas 12 of the 14 patients with mild CHls attained a good recovery (2 were moderately disabled) by the time of study. Although this initial study of brain morphometry after CHI in children was not designed to isolate the contribution of frontal lobe damage to residual disability, further research involving a larger sample is in progress to address this issue.

Measurement of brain compartment volumes in MR using voxel composition calculations.

A computer method was developed for brain compartment volume measurement in MR images. The method is a statistical averaging technique, in which each voxel is viewed as a mixture of adjacent tissues in a measurable proportion. This method is based on sampling representative tissue intensities and then interpolating intermediate intensities. It can automatically correct for volume averaging artifacts occurring in voxels that contain heterogeneous tissues.

The property distance index PD predicts peptides that cross-react with IgE antibodies

Similarities in the sequence and structure of allergens can explain clinically observed cross-reactivities. Distinguishing sequences that bind IgE in patient sera can be used to identify potentially allergenic protein sequences and aid in the design of hypo-allergenic proteins. The property distance index PD, incorporated in our Structural Database of Allergenic Proteins (SDAP, http://fermi.utmb.edu/SDAP/), may identify potentially cross-reactive segments of proteins, based on their similarity to known IgE epitopes. We sought to obtain experimental validation of the PD index as a quantitative predictor of IgE cross-reactivity, by designing peptide variants with predetermined PD scores relative to three linear IgE epitopes of Jun a 1, the dominant allergen from mountain cedar pollen. For each of the three epitopes, 60 peptides were designed with increasing PD values (decreasing physicochemical similarity) to the starting sequence. The peptides synthesized on a derivatized cellulose membrane were probed with sera from patients who were allergic to Jun a 1, and the experimental data were interpreted with a PD classification method. Peptides with low PD values relative to a given epitope were more likely to bind IgE from the sera than were those with PD values larger than 6. Control sequences, with PD values between 18 and 20 to all the three epitopes, did not bind patient IgE, thus validating our procedure for identifying negative control peptides. The PD index is a statistically validated method to detect discrete regions of proteins that have a high probability of cross-reacting with IgE from allergic patients.