The primary motor cortex (M1) is thought to control movements of different body parts from somatotopically organized cortical territories. Electrical stimulation suggests, however, that territories controlling different fingers overlap. Such overlap might be artifactual or else might indicate that activation of M1 to produce a finger movement occurs over a more widespread cortical area than usually assumed. These possibilities were distinguished in monkeys moving different fingers. Recordings showed that single M1 neurons were active with movements of different fingers. Neuronal populations active with movements of different fingers overlapped extensively. Control of any finger movement thus appears to utilize a population of neurons distributed throughout the M1 hand area rather than a somatotopically segregated population.

Abstract. Treatment planning for high precision radiotherapy of head and neck (H&N) cancer patients requires accurate delineation of many structures and lymph node regions. Manual contouring is tedious and suffers from large inter-and intra-rater variability. To reduce manual labor, we have developed a fully automated, atlas-based method for H&N CT image segmentation that employs a novel hierarchical atlas registration approach. This registration strategy makes use of object shape information in the atlas to help improve the registration efficiency and robustness while still being able to account for large inter-subject shape differences. Validation results showed that our method provides accurate segmentation for many structures despite difficulties presented by real clinical data. Comparison of two different atlas selection strategies is also reported.

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Spleen cells obtained from mice immunized with partially purified human coagulation Factor V were fused with NS-1 mouse myeloma cells, and hybrids were selected. Culture media were screened for anti-Factor V activity, and an antibodypositive clone was obtained and passaged as an ascites tumor in mice. The ascitic fluid from the hybridoma-bearing mouse could be diluted 1:106 before losing reactivity in an anti-Factor V radioimmunoassay. When immobilized on agarose, the monoclonal antibody quantitatively removed Factor V activity from human plasma. Factor V activity could be eluted with 1.2 M NaCI at pH 6.5. Homogeneous Factor V was isolated by chromatography of barium citrate-adsorbed, polyethylene glycol 6000 precipitated plasma on the antibody column followed by chromatography on phenyl-Sepharose. The isolated Factor V exhibited a single band upon gel electrophoresis in sodium dodecyl sulfate with an apparent Mr comparable to that of bovine Factor V (330,000). Upon exposure to thrombin, the activity of Factor V increased 53-fold -when measured in Factor V-deficient plasma. This increased activity was associated with discrete proteolytic cleavages of the parent molecule.Blood coagulation Factor V as a functional entity was first described in 1947 by Owren (1). Studies by Owren (2), Ware and Seegers (3), and Murphy and Seegers (4) identified Factor V as one of the components essential for the rapid conversion of prothrombin to the blood clotting enzyme thrombin. Subsequent work indicated that Factor V functions as a cofactor, in concert with phospholipid (or platelets) and Ca2+, to promote the proteolytic activation of prothrombin by Factor Xa (5-12). In addition, several studies (2,6,8,(13)(14)(15)(16)(17) indicated that Factor V itselfis converted by thrombin to the more active form, Factor Va.Our knowledge concerning the function and molecular properties of Factor V expanded after the isolation of the bovine protein (18,19). Bovine Factor V is a high molecular weight (Mr = 330,000), single-chain precursor of Factor Va (18). Once activated by thrombin-catalyzed proteolytic cleavage (19,20), Factor Va enhances (in the presence of Ca2' and phospholipid) the activity ofFactor Xa toward prothrombin by at least 4 orders of magnitude (21,22). Studies of Factor V binding properties with regard to Ca2+ (23), phospholipid (24), and platelets (25) have indicated that Factor Va not only greatly augments rates of prothrombin activation but also can be viewed as a plateletbound receptor for Factor Xa in vivo (25,26).Although bovine Factor V has been characterized reasonably well, the properties of human Factor V have not been elucidated. In our hands, even when substantially modified, procedures that readily permit the isolation of the bovine protein do not yield electrophoretically homogeneous, undegraded human Factor V. Consequently, efforts were undertaken, using partially purified human Factor V, to generate a monoclonal hybridoma antibody for use as a tool in studying the human protein. An antibody was produce...

The efficacy of [14C]glucose molecules labeled in various positions as tracers of regional cerebral glucose utilization (rCMRGlc) was examined in rats. Arteriovenous differences of different [14C]-glucose species and 14CO2 were measured across brain to determine the relative rates of 14CO2 loss. As anticipated, 14CO2 evolution decreased in the order: [U-14C]glucose greater than [2-14C]glucose greater than [1-14C]glucose greater than [6-14C]glucose. Release of 14CO2 from [6-14C]glucose was undetectable at 5 min and barely detectable at 10 min, and release from [1-14C]glucose, which includes the pentose phosphate pathway, was only slightly greater. rCMRGlc was measured with [1-14C]-,[2-14C]-, or [6-14C]glucose in 5-min experiments. The results of [1-14C]- and [6-14C]glucose were indistinguishable; no difference due to the activity of the pentose phosphate pathway was found. Both [1-14C]- and [6-14C]-glucose gave values similar to, but on the whole slightly higher than, [2-14C]glucose. It was concluded that when knowledge of total rCMRGlc is required, [6-14C]glucose is the labeled substrate of choice. When the experimental objective is measurement of energy metabolism, use of [1-14C]glucose avoids inclusion of the nonenergy-yielding pentose phosphate pathway.

Crystals of alpha-chymotrypsin (CHT) at equilibrium in solutions of 2.0 M guanidine hydrochloride and 3.0 M urea at pH 3.6 were prepared, three-dimensional X-ray intensities were measured, and difference electron-density maps were calculated and examined. The guanidine hydrochloride derivative displayed changes occurring exclusively on the surface of the protein. The difference peaks represented mostly small changes in various protein surface groups and in the adjacent solvent regions, and some displayed convincing evidence of binding of the guanidinium ion to the protein. The urea difference map likewise showed that changes had occurred on the surface of the protein, but also that numerous changes in the structure occurred in the hydrophobic interior of the CHT molecule. Further, the urea difference map contained evidence for two kinds of interactions of urea with protein groups. There are examples of bound urea either causing or accompanying structural changes and examples of urea binding with no accompanying changes to the protein. Examples of both kinds of binding were observed in both the surface regions and in the hydrophobic interior of the molecule. From an examination of these two derivatives, it is clear that guanidine hydrochloride and urea unfold proteins by different mechanisms.

Regional cerebral glucose influx was measured using quantitative autoradiography after the intravenous infusion of [2-14C]glucose for a period of 10 or 20 s. Glucose influx varied considerably among structures over an almost threefold range. When compared with rates of regional glucose utilization, a significant correlation by region was found between glucose influx and utilization, demonstrating that the glucose supply to individual cerebral structures is closely matched to their metabolic needs.

Electrophysiological data from the rodent whisker/barrel cortex indicate that GABAergic, presumed inhibitory, neurons respond more vigorously to stimulation than glutamatergic, presumed excitatory, cells. However, these data represent very small neuronal samples in restrained, anesthetized, or narcotized animals or in cortical slices. Histochemical data from primate visual cortex, stained for the mitochondrial enzyme cytochrome oxidase (CO) and for GABA, show that GABAergic neurons are more highly reactive for CO than glutamatergic cells, indicating that inhibitory neurons are chronically more active than excitatory neurons but leaving doubt about the short-term stimulus dependence of this activation. Taken together, these results suggest that highly active inhibitory neurons powerfully influence relatively inactive excitatory cells but do not demonstrate directly the relative activities of excitatory and inhibitory neurons in the cortex during normal behavior.We used a novel double-labeling technique to approach the issue of excitatory and inhibitory neuronal activation during behavior. Our technique combines high-resolution 2-deoxyglucose (2DG), immunohistochemical staining for neurotransmitterspecific antibodies, and automated image analysis to collect the data. We find that putative inhibitory neurons in barrel cortex of behaving animals are, on average, much more heavily 2DG-labeled than presumed excitatory cells, a pattern not seen in animals anesthetized at the time of 2DG injection. This metabolic activation is dependent specifically on sensory inputs from the whiskers, because acute trimming of most whiskers greatly reduces 2DG labeling in both cell classes in columns corresponding to trimmed whiskers. Our results provide confirmation of the active GABAergic cell hypothesis suggested by CO and single-unit data. We conclude that strong activation of inhibitory cortical neurons must confer selective advantages that compensate for its inherent energy inefficiency.