An open-label trial of the PPAR-gamma ligand rosiglitazone for active ulcerative colitis.

OBJECTIVES: Previous research has demonstrated that ligands for the gamma subtype of peroxisome proliferator-activated receptors (PPARs) reduce inflammation in two different murine models of colitis. This study was designed to examine the potential efficacy of rosiglitazone, a ligand for the gamma subtype of PPARs, as a therapy for active ulcerative colitis. METHODS: Fifteen patients with mild to moderately active ulcerative colitis despite therapy with 5-aminosalicylic acid compounds were enrolled in an open-label study of rosiglitazone (4 mg b.i.d. p.o.) for 12 wk. Thirteen of 15 patients were receiving concomitant therapy with corticosteroids and/or immunomodulator medications. Disease activity was measured with the Disease Activity Index. RESULTS: After 12 wk of therapy, four patients (27%) had achieved clinical remission, of whom three (20%) also had an endoscopic remission. Four additional patients (27%) had a clinical response without achieving remission. Two patients were hospitalized with worsened disease activity, and one patient was withdrawn for nephrotic syndrome. CONCLUSIONS: These data suggest that ligands for the gamma subtype of PPARs may represent a novel therapy for ulcerative colitis. A double blind, placebo-controlled, randomized trial is warranted.

Identification of an outer segment targeting signal in the COOH terminus of rhodopsin using transgenic Xenopus laevis.

Mislocalization of the photopigment rhodopsin may be involved in the pathology of certain inherited retinal degenerative diseases. Here, we have elucidated rhodopsin's targeting signal which is responsible for its polarized distribution to the rod outer segment (ROS). Various green fluorescent protein (GFP)/rhodopsin COOH-terminal fusion proteins were expressed specifically in the major red rod photoreceptors of transgenic Xenopus laevis under the control of the Xenopus opsin promoter. The fusion proteins were targeted to membranes via lipid modifications (palmitoylation and myristoylation) as opposed to membrane spanning domains. Membrane association was found to be necessary but not sufficient for efficient ROS localization. A GFP fusion protein containing only the cytoplasmic COOH-terminal 44 amino acids of Xenopus rhodopsin localized exclusively to ROS membranes. Chimeras between rhodopsin and alpha adrenergic receptor COOH-terminal sequences further refined rhodopsin's ROS localization signal to its distal eight amino acids. Mutations/deletions of this region resulted in partial delocalization of the fusion proteins to rod inner segment (RIS) membranes. The targeting and transport of endogenous wild-type rhodopsin was unaffected by the presence of mislocalized GFP fusion proteins.

Cochlear nerve projections to the small cell shell of the cochlear nucleus: the neuroanatomy of extremely thin sensory axons.

Labeling cochlear nerve fibers in the inner ear of chinchillas with biotinylated dextran polyamine was used to trace the thin fibers (Type II), which likely innervate outer hair cells. These axons, 0. 1-0.5 microm in diameter, were distinguished from the thicker Type I, fibers innervating inner hair cells, and traced to small-cell clusters in the cochlear nucleus. This study provided two major new insights into the outer hair cell connections in the cochlear nucleus and the potential significance of very thin axons and synaptic nests, which are widespread in the CNS. 1) EM serial reconstructions of labeled and unlabeled material revealed that Type II axons rarely formed synapses with conventional features (vesicles gathered at junctions). Rather, their endings contained arrays of endoplasmic reticulum and small spherical vesicles without junctions. 2) Type II axons projected predominantly to synaptic nests, where they contacted other endings and dendrites of local interneurons (small stellate and mitt cells, but not granule cells). Synaptic nests lacked intrinsic glia and, presumably, their high-affinity amino acid transporters. As functional units, nests and their Type II inputs from outer hair cells may contribute to an analog processing mode, which is slower, more diffuse, longer-lasting, and potentially more plastic than the digital processors addressed by inner hair cells.

Therapeutic potential of infliximab in inflammatory bowel disease.

Traditional treatment approaches for patients with inflammatory bowel disease involve 1) pharmacologic management with aminosalicylates, corticosteroids, immunomodulatory agents, and antibiotics; and 2) nonpharmacologic management by surgical and dietary means. Complications and inadequate responses to current treatment modalities have resulted in the development of new and more specific biologic therapies. The proinflammatory cytokine tumor necrosis factor alpha (TNF alpha) is present in elevated concentrations in patients with inflammatory bowel disease; therefore, it has been targeted for treatment approaches. Infliximab is a chimeric, monoclonal antibody to TNF alpha developed to inhibit the interaction of TNF alpha with its receptor, to result in amelioration of inflammation. This biologic agent has received the most attention and is the most studied of those for the treatment of patients with Crohn's disease. This article reviews the symptoms of inflammatory bowel disease, the traditional treatment approaches used, the role of TNF alpha in disease pathogenesis, and the value of infliximab in treatment.

Purkinje-like cells in rat cochlear nucleus.

A unique class of cells, strongly immunopositive for anti-calbindin D-28 kDa was observed in and near the cochlear nucleus of young adult, male Sprague-Dawley rats. These cells are present in small numbers which are highly variable across animals and inconstant in position. They are preferentially located in the dorsal cochlear nucleus, with occasional examples being present in the ventral cochlear nucleus, as well as in adjacent brainstem locations. They have been referred to in other studies as displaced Purkinje cells or 'Purkinje cell-like cells', and are here designated 'Purkinje-like cells' (PLCs). PLCs have relatively large cell bodies, with thick, heavily spined dendrites, and are typically situated in an immediately subpial position. The dendritic arborization extends into the interior of the nucleus, away from the pial surface, a trajectory opposite in direction to that of the cerebellar Purkinje cells. The intense immunoreactivity exhibited by PLC somata and dendrites when treated with antiserum directed against calbindin is equivalent to that of cerebellar Purkinje cells, and markedly stronger than that of most other cell populations of the cochlear nucleus. However, in tissue treated with anti-parvalbumin, which also strongly labels cerebellar Purkinje cell somata and dendrites, PLC labeling, when present, is relatively weak, limited to the cell bodies and only the base of the dendrites of PLCs, indicating non-equivalence of the two cell types. In addition, the intensity of calbindin immunostaining in the PLCs appears to be more sensitive to glutaraldehyde in any of the fixative solutions than that seen in cerebellar Purkinje cells in the same sections. Of the cell types of the cochlear nucleus, the cartwheel cells would appear to be the most similar to the PLCs on morphological and immunocytochemical grounds. However, the subpial position and average somal dimensions of the PLCs, as well as the relatively modest immunoreactivity of the cartwheel cells for calbindin, rather clearly differentiate the PLCs from this class of neurons. The results of the present study suggest that the PLCs of the cochlear nucleus, although they may arise developmentally as ectopic cerebellar Purkinje cells and maintain certain Purkinje cell characteristics, represent a distinct neuronal cell type in the adult rat cochlear nucleus, exhibiting incomplete overlap of fixation, immunocytochemical and morphological characteristics with both cartwheel cells of the cochlear nucleus and cerebellar Purkinje cells.

Synaptic microcircuitry of bipolar and amacrine cells with serotonin-like immunoreactivity in the retina of the turtle, Pseudemys scripta elegans.

Although serotonin is thought to be a neurotransmitter in a number of retinal systems, much of the precise synaptic connectivity of serotonergic neurons is unknown. To address this issue, we used an antiserum directed against serotonin to label serotonergic bipolar and amacrine cells in the turtle retina. Light-microscopic analysis of labeled amacrine and bipolar cells indicated that both had bistratified dendritic arborizations primarily in stratum 1 and in strata 4/5 of the inner plexiform layer. Ultrastructural analysis of the neurocircuitry of these cells indicated that the processes of labeled bipolar cells in the outer plexiform layer made basal junction contacts with photoreceptor terminals. Only in rare instances did labeled bipolar cells processes invaginate near photoreceptor ribbon synapses. Processes of labeled bipolar cells received both conventional and small ribbon synaptic contacts in the outer plexiform layer. Bipolar cell processes in stratum 1 of the inner plexiform layer synapsed onto either amacrine/amacrine or amacrine/ganglion cell dyads, and made rare ribbon synaptic contacts onto labeled amacrine cell processes. Synaptic inputs to serotonergic bipolar cells in stratum 1 were from unlabeled bipolar and amacrine cells. Bipolar cell contacts in strata 4/5 were similar to those in stratum 1, but were fewer in number and no bipolar cell inputs were seen. Labeled amacrine cell output in both strata was onto other unlabeled amacrine cells and ganglion cells; but synaptic outputs to unlabeled bipolar cells were only seen in strata 4/5. In both strata 1 and 4/5, synaptic inputs to labeled amacrine cells were from both unlabeled amacrine cells and labeled bipolar cells. The serotonergic amacrine cells had many more synaptic interactions in stratum 1 than in strata 4/5 which supports the role of serotonergic bipolar cells in the OFF pathway of retinal processing. Interactions between serotonergic bipolar and amacrine cells may play an important role in visual processing.

Localization of GABA- and GAD-like immunoreactivity in the turtle retina.

gamma-aminobutyric acid (GABA) has been reported to be an important neurotransmitter in the retinas of many species. This immunocytochemical study detailed the localization of antigens resembling GABA and glutamic acid decarboxylase (GAD, an enzyme involved in the synthesis of GABA), in retinal neurons in the turtle, Pseudemys scripta elegans. GABA-like immunoreactivity was present within somata in the inner and outer regions of the inner nuclear layer, within somata in the ganglion cell layer, and in processes in the outer plexiform layer, inner plexiform layer, and ganglion cell axon layer. GAD-like immunoreactivity was found in somata in the inner and outer regions of the inner nuclear layer and in processes in the inner and outer plexiform layers. Cell counts indicated more somata with GABA-like than GAD-like immunoreactivity in the inner nuclear layer. Double-label studies showed that every somata in the inner nuclear layer which had GAD-like immunoreactivity also had GABA-like immunoreactivity, but that many somata had only GABA-like immunoreactivity. The stratification of immunoreactivity within the inner plexiform layer was analyzed using a scanning densitometer. We described the strate within the inner plexiform layer such that S0 represented the inner nuclear layer/inner plexiform layer border and S100 represented the inner plexiform layer/ganglion cell layer border. Analysis of GAD-like labeling yielded seven distinct strata with peak densities at positions S8, S19, S28, S42, S59, S75, and S93. GABA-like labeling provided five distinct strata with peak densities at positions S17, S28, S67, S84, and S95. The strata with peaks of GABA-like immunoreactivity at S17 and S28 were in statistically identical locations to corresponding strata with GAD-like immunoreactivity. The strata with GABA-like immunoreactivity at S67, S84, and S95 did not have statistically identical peaks of correlated GAD-like immunoreactivity, although there were corresponding strata with GAD-like immunoreactivity nearby. Antiserum directed against GABA failed to produce labeled strata at positions corresponding to the strata with GAD-like immunoreactivity at S8 and S42. In summary, our results indicated that the antisera we used, which were directed against GABA and GAD, produced significantly different labeling in the inner nuclear layer, inner plexiform layer, and the ganglion cell body and axon layers of the turtle retina. Until the physiological significance of these differences is resolved, studies employing these markers to investigate the function of GABA in the turtle retina should be interpreted with caution.