Targeted therapy for advanced salivary gland carcinoma based on molecular profiling: results from MyPathway, a phase IIa multiple basket study.

BACKGROUND: Systemic therapy options for salivary cancers are limited. MyPathway (NCT02091141), a phase IIa study, evaluates targeted therapies in non-indicated tumor types with actionable molecular alterations. Here, we present the efficacy and safety results for a subgroup of MyPathway patients with advanced salivary gland cancer (SGC) matched to targeted therapies based on tumor molecular characteristics. PATIENTS AND METHODS: MyPathway is an ongoing, multiple basket, open-label, non-randomized, multi-center study. Patients with advanced SGC received pertuzumab + trastuzumab (HER2 alteration), vismodegib (PTCH-1/SMO mutation), vemurafenib (BRAF V600 mutation), or atezolizumab [high tumor mutational burden (TMB)]. The primary endpoint is the objective response rate (ORR). RESULTS: As of January 15, 2018, 19 patients with SGC were enrolled and treated in MyPathway (15 with HER2 amplification and/or overexpression and one each with a HER2 mutation without amplification or overexpression, PTCH-1 mutation, BRAF mutation, and high TMB). In the 15 patients with HER2 amplification/overexpression (with or without mutations) who were treated with pertuzumab + trastuzumab, 9 had an objective response (1 complete response, 8 partial responses) for an ORR of 60% (9.2 months median response duration). The clinical benefit rate (defined by patients with objective responses or stable disease >4 months) was 67% (10/15), median progression-free survival (PFS) was 8.6 months, and median overall survival was 20.4 months. Stable disease was observed in the patient with a HER2 mutation (pertuzumab + trastuzumab, n = 1/1, PFS 11.0 months), and partial responses in patients with the PTCH-1 mutation (vismodegib, n = 1/1, PFS 14.3 months), BRAF mutation (vemurafenib, n = 1/1, PFS 18.5 months), and high TMB (atezolizumab, n = 1/1, PFS 5.5+ months). No unexpected toxicity occurred. CONCLUSIONS: Overall, 12 of 19 patients (63%) with advanced SGC, treated with chemotherapy-free regimens matched to specific molecular alterations, experienced an objective response. Data from MyPathway suggest that matched targeted therapy for SGC has promising efficacy, supporting molecular profiling in treatment determination.

Effect of docetaxel duration on clinical outcomes: exploratory analysis of CLEOPATRA, a phase III randomized controlled trial.

Background: Combination pertuzumab, trastuzumab, and docetaxel (D) is considered standard first-line treatment of human epidermal growth factor receptor 2 (HER2)-positive metastatic breast cancer. This post hoc, exploratory analysis of CLEOPATRA study data evaluated the clinical effects of D treatment duration within this regimen. The clinical benefits of pertuzumab and trastuzumab by different durations of D treatment were also evaluated. Patients and methods: Patients with HER2-positive metastatic breast cancer received trastuzumab and D plus pertuzumab or placebo. Clinical outcomes were analyzed by the number of D cycles that patients received (<6D, 6D, or >6D). Progression-free survival (PFS) and overall survival (OS) for each treatment arm within each D cycle group were estimated using the Kaplan-Meier approach. Time-dependent, multivariate Cox regression was applied to estimate adjusted hazard ratios (HRs) and 95% confidence intervals (CIs) for HER2-targeted therapy and D cycle groups. Results: Overall, 804 patients received <6D (n = 119), 6D (n = 210), or >6D (n = 475) cycles. After adjusting for pertuzumab benefits versus placebo (PFS HR = 0.61, 95% CI 0.51-0.74, P < 0.0001; OS HR = 0.60, 95% CI, 0.49-0.74, P < 0.0001), >6D versus 6D cycles was not associated with statistically significant improvements in PFS (HR = 0.80, 95% CI 0.63-1.01, P = 0.0640) or OS (HR = 0.88, 95% CI 0.69-1.12, P = 0.3073). Consistent improvements in PFS and OS were observed with pertuzumab versus placebo, irrespective of D duration. The HRs for PFS were 0.395, 0.615, and 0.633 for <6D, 6D, and >6D cycles, respectively (P < 0.05 for all D cycle groups). Corresponding HRs for OS were 0.577, 0.700, and 0.612, respectively (P < 0.05 for <6D and >6D). Conclusions: After accounting for pertuzumab benefits, more than six cycles of D treatment was not associated with significant improvements in either PFS or OS compared with six cycles. The addition of pertuzumab to trastuzumab improved clinical outcomes versus trastuzumab plus placebo, regardless of D treatment duration. ClinicalTrials.gov identifier: NCT00567190.

Incidence and management of diarrhea in patients with HER2-positive breast cancer treated with pertuzumab.

Background: Pertuzumab disrupts heterodimerization between human epidermal growth factor receptor 2 (HER2) and epidermal growth factor receptor (EGFR), HER3, and HER4. Thus, pertuzumab could result in adverse events similar to those observed with EGFR antagonists, such as diarrhea. We report the incidence and severity of diarrhea observed with pertuzumab in the CLEOPATRA, NeoSphere, and TRYPHAENA studies. Patients and methods: Patients (n = 1443) had metastatic [CLEOPATRA (n = 804)] or early-stage breast cancer [NeoSphere (n = 416) and TRYPHAENA (n = 223)]. The incidence and severity of diarrhea were analyzed by treatment received. The incidence of febrile neutropenia concurrent with diarrhea and the effect of pre-existing gastrointestinal comorbidities were also evaluated. Subgroup analyses were carried out using CLEOPATRA data. Results: The incidence of all-grade diarrhea across studies was generally greater for pertuzumab-based treatment, ranging from 28% to 72% (grade 1, 21%-54%; grade 2, 8%-37%; grade 3, 0%-12%; grade 4, 0%). Incidence was highest during the first pertuzumab-containing cycle, decreasing with subsequent cycles. Dose delays or discontinuations due to diarrhea were infrequent, ranging from 0% to 8%. Among pertuzumab-treated patients with diarrhea, 47%-67% received pharmacological intervention, most commonly with loperamide. Overlap between diarrhea and febrile neutropenia was uncommon, ranging from 0% to 11%. No relationship was observed between pre-existing gastrointestinal comorbidities and diarrhea. In CLEOPATRA, patients ≥65 years treated with pertuzumab had a higher incidence of grade 3 diarrhea than patients <65 years (19% versus 8%). All-grade diarrhea occurred at greater frequency among pertuzumab-treated Asian versus white patients with metastatic breast cancer (74% versus 63%); the corresponding rates in the control arm were 53% and 45%, respectively. Conclusions: In both the metastatic and early-stage breast cancer settings, diarrhea was common but manageable for all pertuzumab-containing regimens. Diarrheal episodes were mainly low grade and occurred most often during the first treatment cycle. Diarrheal-related drug delays or discontinuations were uncommon. ClinicalTrials.gov identifiers: NCT00567190 (CLEOPATRA), NCT00545688 (NeoSphere), NCT00976989 (TRYPHAENA).

Apoptosis and delayed degeneration after spinal cord injury in rats and monkeys.

Apoptosis is a morphologically defined form of programmed cell death seen in a variety of circumstances, including immune cell selection, carcinogenesis and development. Apoptosis has very recently been seen after ischemic or traumatic injury to the central nervous system (CNS), suggesting that active cell death as well as passive necrosis may mediate damage after CNS injury. After spinal cord injury (SCI) in the rat, typical post-traumatic necrosis occurred, but in addition, apoptotic cells were found from 6 hours to 3 weeks after injury, especially in the spinal white matter. Apoptotic cells were positive for oligodendrocyte markers. After SCI in monkeys, apoptotic cells were found within remote degenerating fiber tracts. Both secondary degeneration at the site of SCI and the chronic demyelination of tracts away from the injury appear to be due in part to apoptosis. As cytokines have been shown to mediate oligodendrocyte death in vitro, it seems likely that chronic demyelination after CNS injury shares features with chronic degenerative disorders like multiple sclerosis.

Machine intelligence identifies soluble TNFa as a therapeutic target for spinal cord injury.

Traumatic spinal cord injury (SCI) produces a complex syndrome that is expressed across multiple endpoints ranging from molecular and cellular changes to functional behavioral deficits. Effective therapeutic strategies for CNS injury are therefore likely to manifest multi-factorial effects across a broad range of biological and functional outcome measures. Thus, multivariate analytic approaches are needed to capture the linkage between biological and neurobehavioral outcomes. Injury-induced neuroinflammation (NI) presents a particularly challenging therapeutic target, since NI is involved in both degeneration and repair. Here, we used big-data integration and large-scale analytics to examine a large dataset of preclinical efficacy tests combining five different blinded, fully counter-balanced treatment trials for different acute anti-inflammatory treatments for cervical spinal cord injury in rats. Multi-dimensional discovery, using topological data analysis (TDA) and principal components analysis (PCA) revealed that only one showed consistent multidimensional syndromic benefit: intrathecal application of recombinant soluble TNFα receptor 1 (sTNFR1), which showed an inverse-U dose response efficacy. Using the optimal acute dose, we showed that clinically-relevant 90 min delayed treatment profoundly affected multiple biological indices of NI in the first 48 h after injury, including reduction in pro-inflammatory cytokines and gene expression of a coherent complex of acute inflammatory mediators and receptors. Further, a 90 min delayed bolus dose of sTNFR1 reduced the expression of NI markers in the chronic perilesional spinal cord, and consistently improved neurological function over 6 weeks post SCI. These results provide validation of a novel strategy for precision preclinical drug discovery that is likely to improve translation in the difficult landscape of CNS trauma, and confirm the importance of TNFα signaling as a therapeutic target.

Convolutional Neural Network-Based Automated Segmentation of the Spinal Cord and Contusion Injury: Deep Learning Biomarker Correlates of Motor Impairment in Acute Spinal Cord Injury.

BACKGROUND AND PURPOSE: Our aim was to use 2D convolutional neural networks for automatic segmentation of the spinal cord and traumatic contusion injury from axial T2-weighted MR imaging in a cohort of patients with acute spinal cord injury. MATERIALS AND METHODS: Forty-seven patients who underwent 3T MR imaging within 24 hours of spinal cord injury were included. We developed an image-analysis pipeline integrating 2D convolutional neural networks for whole spinal cord and intramedullary spinal cord lesion segmentation. Linear mixed modeling was used to compare test segmentation results between our spinal cord injury convolutional neural network (Brain and Spinal Cord Injury Center segmentation) and current state-of-the-art methods. Volumes of segmented lesions were then used in a linear regression analysis to determine associations with motor scores. RESULTS: Compared with manual labeling, the average test set Dice coefficient for the Brain and Spinal Cord Injury Center segmentation model was 0.93 for spinal cord segmentation versus 0.80 for PropSeg and 0.90 for DeepSeg (both components of the Spinal Cord Toolbox). Linear mixed modeling showed a significant difference between Brain and Spinal Cord Injury Center segmentation compared with PropSeg (P < .001) and DeepSeg (P < .05). Brain and Spinal Cord Injury Center segmentation showed significantly better adaptability to damaged areas compared with PropSeg (P < .001) and DeepSeg (P < .02). The contusion injury volumes based on automated segmentation were significantly associated with motor scores at admission (P = .002) and discharge (P = .009). CONCLUSIONS: Brain and Spinal Cord Injury Center segmentation of the spinal cord compares favorably with available segmentation tools in a population with acute spinal cord injury. Volumes of injury derived from automated lesion segmentation with Brain and Spinal Cord Injury Center segmentation correlate with measures of motor impairment in the acute phase. Targeted convolutional neural network training in acute spinal cord injury enhances algorithm performance for this patient population and provides clinically relevant metrics of cord injury.

Multivariate Analysis of MRI Biomarkers for Predicting Neurologic Impairment in Cervical Spinal Cord Injury.

BACKGROUND AND PURPOSE: Acute markers of spinal cord injury are essential for both diagnostic and prognostic purposes. The goal of this study was to assess the relationship between early MR imaging biomarkers after acute cervical spinal cord injury and to evaluate their predictive validity of neurologic impairment. MATERIALS AND METHODS: We performed a retrospective cohort study of 95 patients with acute spinal cord injury and preoperative MR imaging within 24 hours of injury. The American Spinal Injury Association Impairment Scale was used as our primary outcome measure to define neurologic impairment. We assessed several MR imaging features of injury, including axial grade (Brain and Spinal Injury Center score), sagittal grade, length of injury, maximum canal compromise, and maximum spinal cord compression. Data-driven nonlinear principal component analysis was followed by correlation and optimal-scaled multiple variable regression to predict neurologic impairment. RESULTS: Nonlinear principal component analysis identified 2 clusters of MR imaging variables related to 1) measures of intrinsic cord signal abnormality and 2) measures of extrinsic cord compression. Neurologic impairment was best accounted for by MR imaging measures of intrinsic cord signal abnormality, with axial grade representing the most accurate predictor of short-term impairment, even when correcting for surgical decompression and degree of cord compression. CONCLUSIONS: This study demonstrates the utility of applying nonlinear principal component analysis for defining the relationship between MR imaging biomarkers in a complex clinical syndrome of cervical spinal cord injury. Of the assessed imaging biomarkers, the intrinsic measures of cord signal abnormality were most predictive of neurologic impairment in acute spinal cord injury, highlighting the value of axial T2 MR imaging.

Distribution and morphology of sacral spinal cord neurons innervating pelvic structures in Xenopus laevis.

Relatively little is known about the organization of neural input to pelvic viscera in amphibia. In this study, sacral spinal efferent neurons were labeled in Xenopus laevis frogs by application of horseradish peroxidase (HRP) to the tenth spinal nerve, to pelvic musculature, or to the pelvic nerve. DiI was applied to the pelvic nerve with similar results. Labeled spinal neurons were located in the intermediate gray or in the ventral horn. Neurons in the tenth dorsal root ganglion, but not in the spinal cord, were labeled after application of HRP or DiI to the pudendal nerve. The labeled neurons in the spinal cord intermediate gray were in a position comparable to that of the mammalian sacral parasympathetic nucleus (SPN). Two apparent subdivisions included 1) a medial cluster of cells with mediolaterally oriented dendrites and 2) a lateral group with dorsoventrally oriented dendrites. An intermediate group, not clearly classed with the other two, was also identifiable. In some cases, labeled tenth nerve primary afferents were seen in contact with efferent neurons of the intermediate gray. Labeled neurons in the ventral horn medial to the lateral motor column were small, with dendrites oriented mediolaterally, in a position comparable to that of the mammalian Onuf's nucleus. The peripheral targets of DiI-labeled pelvic nerve axons were the compressor cloaca muscle, cloaca, and bladder. DiI-labeled pudendal nerve axons distributed peripherally to cloacal lip and medial thigh integument. These data suggest that the pudendal nerve in amphibians is purely sensory and that both somatic and autonomic motor axons traverse the pelvic nerve.

A light and electron microscopic analysis of the sacral parasympathetic nucleus after labelling primary afferent and efferent elements with HRP.

Primary afferent input to the cat sacral parasympathetic nucleus (SPN) has been examined by injury filling sacral dorsal roots, ventral roots, or both with horseradish peroxidase (HRP). Appropriate spinal segments were processed for the demonstration of HRP with diaminobenzidine and prepared for sequential light (LM) and electron (EM) microscopy. At the LM level, a large fascicle of primary afferent fibers was observed passing ventrally along the lateral edge of the dorsal horn into the region of the SPN. Varicosities were seen throughout the course of the axons but were particularly abundant within the SPN. Injury filling of the ventral roots with HRP resulted in a Golgi-like labelling of preganglionic neurons and their dendritic arbors, as well as ventral root afferent fibers. Swellings on both dorsal and ventral root afferent axons were observed in close apposition to labelled preganglionic neurons and their dendrites. At the ultrastructural level, afferent terminals were found to contain clear spherical vesicles; 66% of these terminals also contained at least one dense-cored vesicle. Of particular interest was the presence of labelled dorsal and ventral root afferent terminals synapsing on labelled preganglionic neurons. Preganglionic neurons were also postsynaptic to unlabelled terminals containing clear spherical (79.7%) or pleomorphic vesicles (20.3%). These data indicate that preganglionic neurons receive direct input from several sources, and provide the first demonstration of direct input to these cells from sensory fibers in the dorsal and ventral roots. The connections described in the present study provide interesting and, as yet, unexplored possibilities for sensory and autonomic reflex integration.

A comparison of the ultrastructure of substance P and enkephalin-immunoreactive elements in the nucleus of the dorsal lateral funiculus and laminae I and II of the rat spinal cord.

The ultrastructure of substance P (SP)- and enkephalin (ENK)-immunoreactive elements in the nucleus of the dorsal lateral funiculus (NDLF) and in laminae I and II of the spinal cord was examined in the rat using the peroxidate-antiperoxidase (PAP) technique. Electron-microscopic observations were made of a large number of immunolabelled terminals (n = 428; many followed in serial sections), axons, and immunoreactive cell bodies and dendrites which were occasionally encountered. Morphometric analysis was used to describe and compare the fine structural features of immunolabelled elements. Both SP- and ENK-immunoreactive terminals contained clear synaptic vesicles and dense-cored vesicles of similar size but the ENK-immunoreactive profiles contained significantly more dense-cored vesicles than SP-immunolabelled profiles. Both SP- and ENK-immunoreactive profiles in the dorsal laminae of the dorsal horn contacted mainly smaller dendritic elements. Only rarely were axo-axonic interactions noted. The NDLF contains widely scattered cell bodies dispersed within a neuropil which is rich in synaptic complexes and is interdigitated between fascicles of myelinated and unmyelinated axons. Numerous SP- and ENK-immunoreactive profiles were observed in the NDLF, many of which made asymmetric synaptic contacts with NDLF neurons. Although both the dorsal gray and NDLF contain large numbers of SP- and ENK-immunoreactive elements which are similar in morphology in both regions, the NDLF can be distinguished from laminae I and II by a number of criteria, including the nature of the neuropil, principle sources of SP innervation, and the termination patterns of ascending projections.

Differences in synaptic inputs to preganglionic neurons in the dorsal and lateral band subdivisions of the cat sacral parasympathetic nucleus.

In the cat, preganglionic neurons (PGNs) found in the dorsal portion of the sacral parasympathetic nucleus (dorsal band or DB cells) participate in bowel control, while those found along the lateral edge (lateral band or LB cells) influence bladder function. In order to determine whether differences in the synaptic inputs exist between these two populations, HRP was applied to the sacral ventral rootlets of cats, and the S2 cord segment was prepared for sequential light and electron microscopy. When measured with light microscope, the LB somata had greater cross-sectional areas than did the DB cells. Ultrastructurally, the LB cells had a significantly greater percentage of their membrane apposed by synaptic active sites than did the DB cells. Also, the proximal dendrities of the labelled neurons received greater synaptic input than did the somata. No difference was found in the proportion of terminals containing dense cored vesicles (DCVs) when comparing LB and DB somata; however, the LB proximal dendrites had a higher proportion of their surface contacted by DCV-containing terminals than did the DB dendrites. These ultrastructural results offer evidence that these two populations of preganglionic neurons differ with respect to their synaptic input as well as their peripheral targets.

Descending projections from the nucleus raphe obscurus to pudendal motoneurons in the male rat.

Previous physiological and behavioral studies have shown that the nucleus raphe obscurus (nRO) modulates pelvic floor reflex function (Yamanouchi and Kakeyama [1992] Physiol. Behav. 51:575-579; Beattie et al. [1996] Soc. Neurosci. Abstr. 22:722.4; Holmes et al. [1997] Brain Res. 759:197-204). In the present study, small injections of fluorescent tracers were used to investigate direct descending projections from the rostral and caudal portions of the brainstem nRO to retrogradely labeled pudendal motoneurons (MN) in the male rat. The caudal nRO projects into the ventral and lateral funiculi of the spinal cord, with arborizations in the thoracic intermediolateral cell column; in laminae VII, IX, and X of the lumbosacral cord; and in the sacral parasympathetic nucleus (SPN). Many identified external anal sphincter and ischiocavernosus MNs appeared to be in direct apposition with fibers originating from the caudal nRO; and more than half of the bulbospongiosus MNs that were identified appeared to receive such descending input. In addition to the nRO spinal autonomic and pudendal motoneuronal targets, projections were observed to regions of the intermediate gray that contain interneurons organizing the pelvic floor reflexes and to MN pools that are involved in functionally related somatic activities. Finally, several neurons in the lumbar enlargement were labeled retrogradely with FluoroRuby after injections into the nRO and the immediately adjacent reticular formation. Thus, the nRO may be in a position to modulate the coordinated actions of autonomic preganglionic and functionally related skeletal MN activity involved in sexual and eliminative reflex functions.

The relationship of dorsal root afferents to motoneuron somata and dendrites in the adult bullfrog: a light and electron microscopic study using horseradish peroxidase.

The relationship of lumbar dorsal root afferents to lateral motor column motoneurons was studied using anterograde injury filling of dorsal roots and retrograde injury filling of ventral roots with horseradish peroxidase. At the light microscopic level, horseradish peroxidase labelled dorsal root axons were observed to separate into a medial division of large diameter axons which enter the dorsal funiculus and a lateral division of small diameter axons which form a compact bundle in the dorsolateral funiculus which may be homologous to the mammalian tract of Lissauer. Within the spinal gray, primary afferents terminate in two distinct regions. The more ventral of these terminal fields, which receives collaterals of primary afferent axons in the dorsal funiculus, overlaps the dendritic arborizations of the lateral motor column motoneurons. Some axons leave the ventral terminal field to enter the dorsal lateral motor column. Here they terminate on the primary dendrites and somata of lateral motor column motoneurons. At the electron microscopic level, labelled primary afferent terminals were seen to synapse upon lateral motor column motoneuron dendrites as well as upon the somata of dorsally positioned lateral motor column motoneurons. These terminals contain small spherical vesicles and occasional dense-cored vesicles. The synaptic specializations are characterized by a small amount of postsynaptic material. The lateral motor column may be divided into dorsal and ventral portions on the basis of the primary afferent distribution and this is in accord with functional, physiological and developmental data.

A sensitive and reliable locomotor rating scale for open field testing in rats.

Behavioral assessment after spinal cord contusion has long focused on open field locomotion using modifications of a rating scale developed by Tarlov and Klinger (1954). However, on-going modifications by several groups have made interlaboratory comparison of locomotor outcome measures difficult. The purpose of the present study was to develop an efficient, expanded, and unambiguous locomotor rating scale to standardize locomotor outcome measures across laboratories. Adult rats (n = 85) were contused at T7-9 cord level with an electromagnetic or weight drop device. Locomotor behavior was evaluated before injury, on the first or second postoperative day, and then for up to 10 weeks. Scoring categories and attributes were identified, operationally defined, and ranked based on the observed sequence of locomotor recovery patterns. These categories formed the Basso, Beattie, Bresnahan (BBB) Locomotor Rating Scale. The data indicate that the BBB scale is a valid and predictive measure of locomotor recovery able to distinguish behavioral outcomes due to different injuries and to predict anatomical alterations at the lesion center. Interrater reliability tests indicate that examiners with widely varying behavioral testing experience can apply the scale consistently and obtain similar scores. The BBB Locomotor Rating Scale offers investigators a more discriminating measure of behavioral outcome to evaluate treatments after spinal cord injury.

Review of current evidence for apoptosis after spinal cord injury.

The initial mechanical tissue disruption of spinal cord injury (SCI) is followed by a period of secondary injury that increases the size of the lesion. The secondary injury has long been thought to be due to the continuation of cellular destruction through necrotic (or passive) cell death. Recent evidence from brain injury and ischemia suggested that cellular apoptosis, an active form of programmed cell death seen during development, could play a role in CNS injury in adulthood. Here, we review the evidence that apoptosis may be important in the pathophysiology of SCI. There is now strong morphological and biochemical evidence from a number of laboratories demonstrating the presence of apoptosis after SCI. Apoptosis occurs in populations of neurons, oligodendrocytes, microglia, and, perhaps, astrocytes. The death of oligodendrocytes in white matter tracts continues for many weeks after injury and may contribute to post-injury demyelination. The mediators of apoptosis after SCI are not well understood, but there is a close relationship between microglia and dying oligodendrocytes, suggesting that microglial activation may be involved. There is also evidence for the activation of important intracellular pathways known to be involved in apoptosis in other cells and systems. For example, some members of the caspase family of cysteine proteases are activated after SCI. It appears that the evolution of the lesion after SCI involves both necrosis and apoptosis. It is likely that better understanding of apoptosis after SCI will lead to novel strategies for therapeutic interventions that can diminish secondary injury.

Three-dimensional computer-assisted analysis of graded contusion lesions in the spinal cord of the rat.

Histological analysis of spinal cord injury in experimental animals has focused primarily on the microanatomy of damaged tissue. The current study presents an analysis of the three-dimensional structure of lesion sites in the spinal cord of rats contused with an injury device which produces consistent lesions. Three levels of injury were produced by systematically varying the cord displacement and the duration of the displacement during impact. The resulting groups of subjects exhibited mild, moderate, and severe neurological deficits. Comparisons of equivalent mild impacts made at thoracic versus lumbar spinal cord levels were also made. The results indicate that the overall shape of the lesions is generally biconical, with extensions in the base of the dorsal funiculus, irrespective of the degree of damage or the spinal level of the injury. Lower displacement injuries yielded shorter lesions rostrocaudally with less spread into the white matter. Similar impacts in the lumbar versus thoracic spinal cord produced shorter, more truncated lesion sites at lumbar levels with less involvement of the white matter than in the thoracic lesions. Three-dimensional analyses can can provide additional information about the lesion beyond that available from conventional histopathological measures. Such information could be useful in assessing the results of posttraumatic manipulations which are directed at reducing tissue damage or tissue replacement via transplantation.

Spinal cord injury produced by consistent mechanical displacement of the cord in rats: behavioral and histologic analysis.

We examined the ability of an electromechanical device to produce consistent and incomplete thoracic (T9) spinal cord injuries in rats by brief displacement (Dspl) of the exposed dural surface. Open field walking, inclined plane, grid walking, and footprint analysis, and a determination of the percentage of tissue spared at the lesion center were used to assess chronic outcome (6 weeks postinjury). Laminectomy control animals showed no evidence of a functional deficit or histologic lesion. Complete spinal cord transections in normal rats and in a group of animals previously injured (1.1 mm Dspl) and allowed to recover resulted in complete loss of hindlimb function, demonstrating an important functional role for the remaining spared fibers at the lesion site. Consistent spinal cord displacements (0.80 mm, 0.95 mm, and 1.10 mm) resulted in behavioral groups with low outcome variability over a narrow range of incomplete recovery of neurologic function. Significant behavioral (open field walking, inclined plane, and grid walking) and histologic differences were found between the control and Dspl groups and between the 0.80 mm and 1.10 mm Dspl groups. Significant correlations were observed among the injury parameters, behavioral, and histologic scores. Open field walking and inclined plane performance were sensitive indicators of both the early and late phases of neurologic recovery. Grid walking was most useful in animals with small chronic residual deficits. The footprint analysis resulted in less significant correlations and differences between the behavioral groups than the other outcome measures. This may result from a relatively narrow range of sensitivity (open field walking scores between 3.3 and 4.0) and increased variability within the groups.

External anal sphincter hyperreflexia following spinal transection in the rat.

In the present study, long-term and short-term rat preparations were used to develop a model for investigating external anal sphincter (EAS) reflexes in intact and spinal cord-injured (SCI) rats. In this model, EAS distension with an external probe elicits reflex contractions of the EAS in intact, unanesthetized animals. At 2 h after spinal cord transection, none of the lesioned animals displayed EAS EMG activity. In fact, once distended, the EAS was incapable of maintaining closure of the anal orifice. Over a period of 4 days, spinalized animals developed a hyperreflexia of the EAS response. By 48 h, the rectified, integrated EAS EMG was significantly elevated in comparison with nonlesioned controls (EAS hyperreflexia). In addition, the duration of the EAS EMG bursts in response to sphincter distension had significantly increased. At 6 weeks after injury, the EAS was significantly hyperreflexic as measured by EMG burst duration and burst area. As with intact animals, posttransection EAS reflexes were highly anesthesia sensitive. These studies indicate that (1) brief distension of the anal orifice is sufficient to evoke a physiologically relevant reflexive activation of the EAS in the rat, (2) the 2- to 24-h postinjury areflexia observed in these experiments may be a suitable model for the study of spinal shock, and (3) the observed EAS hyperreflexia after chronic SCI may represent the permanent effects of removing descending inhibitory circuits and segmental plasticity, making this reflex an appropriate measure of defecatory dysfunction after spinal cord injury.