bluuebunny/crossref_metadata_2025 huggingface dataset:
Characterization of CART neurons in the rat and human hypothalamus
Authors: Elias Carol F. , Lee Charlotte E. , Kelly Joseph F. , Ahima Rexford S. , Kuhar Michael , Saper Clifford B. , Elmquist Joel K.
Published: August 2002

Cocaine‐ and amphetamine‐regulated transcript (CART) is a recently described neuropeptide widely expressed in the rat brain. CART mRNA and peptides are found in hypothalamic sites such as the paraventricular nucleus (PVH), the supraoptic nucleus (SON), the lateral hypothalamic area (LHA), the dorsomedial nucleus of the hypothalamus (DMH), the arcuate nucleus (Arc), the periventricular nucleus (Pe), and the ventral premammillary nucleus (PMV). Intracerebroventricular administration of recombinant CART peptide decreases food intake and CART mRNA levels in the Arc are regulated by leptin. Leptin administration induces Fos expression in hypothalamic CART neurons in the PVH, the DMH, the Arc, and the PMV. In the current study, we used double label in situ hybridization histochemistry to investigate the potential direct action of leptin on hypothalamic CART neurons and to define the chemical identity of the hypothalamic CART neurons in the rat brain. We found that CART neurons in the Arc, DMH, and PMV express long form leptin‐receptor mRNA, and the suppressor of cytokine signaling‐3 (SOCS‐3) mRNA after an acute dose of intravenous leptin. We also found that CART neurons in the parvicellular PVH, in the DMH and in the posterior Pe coexpress thyrotropin‐releasing hormone (TRH) mRNA. CART neurons in the magnocellular PVH and in the SON coexpress dynorphin (DYN), and CART cell bodies in the LHA and in the posterior Pe coexpress melanin‐concentrating hormone (MCH) and glutamic acid decarboxylase (GAD‐67) mRNA. In the Arc, a few CART neurons coexpress neurotensin (NT) mRNA. In addition, we examined the distribution of CART immunoreactivity in the human hypothalamus. We found CART cell bodies in the PVH, in the SON, in the LHA, in the Arc (infundibular nucleus) and in the DMH. We also observed CART fibers throughout the hypothalamus, in the bed nucleus of the stria terminalis, and in the amygdala. Our results indicate that leptin directly acts on CART neurons in distinct nuclei of the rat hypothalamus. Furthermore, hypothalamic CART neurons coexpress neuropeptides involved in energy homeostasis, including MCH, TRH, DYN, and NT. The distribution of CART cell bodies and fibers in the human hypothalamus indicates that CART may also play a role in the regulation of energy homeostasis in humans. J. Comp. Neurol. 432:1–19, 2001. © 2001 Wiley‐Liss, Inc.
Neuropeptide expression in rat paraventricular hypothalamic neurons that project to the spinal cord
Authors: Hallbeck Martin , Larhammar Dan , Blomqvist Anders
Published: August 2002

The paraventricular hypothalamic nucleus (PVH) exerts many of its regulatory functions through projections to spinal cord neurons that control autonomic and sensory functions. By using in situ hybridization histochemistry in combination with retrograde tract tracing, we analyzed the peptide expression among neurons in the rat PVH that send axons to the spinal cord. Projection neurons were labeled by immunohistochemical detection of retrogradely transported cholera toxin subunit B, and radiolabeled long riboprobes were used to identify neurons containing dynorphin, enkephalin, or oxytocin mRNA. Of the spinally projecting neurons in the PVH, approximately 40% expressed dynorphin mRNA, 40% expressed oxytocin mRNA, and 20% expressed enkephalin mRNA. Taken together with our previous findings on the distribution of vasopressin‐expressing neurons in the PVH (Hallbeck and Blomqvist [1999] J. Comp. Neurol. 411:201–211), the results demonstrated that the different PVH subdivisions display distinct peptide expression patterns among the spinal cord–projecting neurons. Thus, the lateral parvocellular subdivision contained large numbers of spinal cord–projecting neurons that express any of the four investigated peptides, whereas the ventral part of the medial parvocellular subdivision displayed a strong preponderance for dynorphin‐ and vasopressin‐expressing cells. The dorsal parvocellular subdivision almost exclusively contained dynorphin‐ and oxytocin‐expressing spinal cord–projecting neurons. This parcellation of the peptide‐expressing neurons suggested a functional diversity among the spinal cord–projecting subdivisions of the PVH that provide an anatomic basis for its various and distinct influences on autonomic and sensory processing at the spinal level. J. Comp. Neurol. 433:222–238, 2001. © 2001 Wiley‐Liss, Inc.
Radial glia development in the mouse olfactory bulb
Authors: Puche Adam C. , Shipley Michael T.
Published: August 2002

Radial glia are critical for cell migration and lamination of the cortex. In most developing cortical structures, radial glia, as their name suggests, extend processes from the ventricle to the pia in regular parallel arrangements. However, immunohistochemical labeling from several laboratories suggests that radial glia have a more branched morphology in the olfactory bulb. To investigate the morphology of radial glia in the mouse olfactory bulb we (1) labeled radial glia and olfactory receptor neuron axons at 24‐hour intervals by immunohistochemistry; and (2) developed a novel method of generating and applying “nanocrystals” of 1,1′‐dioctadecyl‐3,3,3′,3′‐ tetramethylindocarbocyanine perchlorate (DiI) to the ventricle surface such that the processes of single olfactory bulb radial glia are labeled in the embryonic olfactory bulb. We examined the structure and interactions of radial glia with ingrowing olfactory receptor neuron (ORN) axons in late embryonic olfactory bulb development. These results showed that olfactory bulb radial glia do not form straight parallel structures as do radial glia in the neocortex but rather have a convoluted trajectory from the ventricle to the bulb surface. Moreover, olfactory bulb radial glia consistently extend tangential branches at the level of the internal plexiform layer. Beginning at embryonic day 17.5, two types of radial glia can be distinguished: type I radial glia have a process that extends from the ventricle into the glomerular layer. These apical processes form highly restricted tufts, or “glial glomeruli” at the same time that ORN axons are forming “axonal glomeruli.” In type II radial glia the apical process does not enter the glomerular layer but instead ramifies within the external plexiform layer. The tight spatiotemporal relationship between the glomerulization of radial glia processes and ORN axons during development suggest that radial glia processes could play a role in the formation and/or stabilization of mammalian glomeruli. J. Comp. Neurol. 434:1–12, 2001. © 2001 Wiley‐Liss, Inc.
Plasticity of neurohypophysial terminals with increased hormonal release during dehydration: Ultrastructural and biochemical analyses
Authors: Miyata Seiji , Takamatsu Hiroyuki , Maekawa Shohei , Matsumoto Naoko , Watanabe Kazutada , Kiyohara Toshikazu , Hatton Glenn I.
Published: August 2002

Arginine vasopressin‐ (AVP) and oxytocin‐ (OXT) secreting magnocellular neurons undergo gross structural changes with chronic physiological stimulation. Here, we investigated subcellular aspects of plasticity in rat neurohypophysial terminals during dehydration. Ultrastructural analyses demonstrated that chronic dehydration by 2% NaCl drinking for 7 days significantly decreased the numbers of neurosecretory granules and microvesicles but not the numbers of mitochondria. Moreover, in dehydrated rats, terminals making neurovascular contacts enlarged, whereas terminals in apposition to astrocytes, i.e., neuroglial contacts, became smaller. Western blot analyses demonstrated significant decreases in the levels of F3 and Thy‐1 together with those of AVP‐ and OXT‐neurophysin, but the levels of synaptophysin, SNAP‐25, and GAP‐43 were unchanged. Both F3 and Thy‐1 were recovered in the buffer‐insoluble pellet, and phosphatidyl inositol‐specific phospholipase C treatment released both molecules from the crude membrane fraction, indicating that they are attached to terminal membranes by glycosylphosphatidyl inositol anchors. Confocal microscopic observations demonstrated that F3 colocalized with Thy‐1 in the same terminals of magnocellular neurons. In contrast, the level of calretinin, a Ca2+ binding protein was significantly increased with chronic dehydration. Thus, the present results suggest that enhancement of neurovascular contacts results from rearrangement of terminal‐astrocyte and terminal‐vessel contacts rather than enlargement or sprouting of magnocellular terminals themselves. The down‐regulation of F3 and Thy‐1 may contribute to enhancement of neurovascular contacts that accompany increased peptide release during dehydration. J. Comp. Neurol. 434:413–427, 2001. © 2001 Wiley‐Liss, Inc.
Spatiotemporal pattern of nicotinamide adenine dinucleotide phosphate‐diaphorase reactivity in the developing central nervous system of premetamorphic Xenopus laevis tadpoles
Authors: McLean David L. , Sillar Keith T.
Published: August 2002

We have catalogued the progressive appearance of putative nitric oxide synthase (NOS)‐containing neurons in the developing central nervous system (CNS) of Xenopus laevis. Xenopus embryos and larvae were processed in wholemount and in cross section using nicotinamide adenine dinucleotide phosphate‐diaphorase (NADPH‐d) histochemistry as a marker for NOS within the CNS. The temporal sequence of NADPH‐d reactivity identified discrete groups and subgroups of neurons in the forebrain, midbrain, and hindbrain on the basis of their morphology, location, and order of appearance during development. A proportion of these groups of neurons appeared to be important in sensory processing and motor control. Staining also appeared at specific stages in the spinal cord, the retina, and the skin. After the appearance of labelling, NADPH‐d reactivity continued in each of the cell groups throughout the stages examined. We found no evidence for staining that subsequently disappeared at later stages in any cell group, indicating a persistent rather than transient role for NO in the Xenopus tadpole CNS. These results are discussed in light of recent findings on possible roles for NADPH‐d‐positive cell groups within the developing motor circuitry. J. Comp. Neurol. 437:350–362, 2001. © 2001 Wiley‐Liss, Inc.
5‐HT3‐receptor subunits A and B are co‐expressed in neurons of the dorsal root ganglion
Authors: Morales Marisela , McCollum Nicholas , Kirkness Ewen F.
Published: August 2002

The type 3 serotonin (5‐HT3) receptor is the only ligand‐gated ion channel receptor for serotonin (5‐HT). Many pharmacological, behavioral, and electrophysiological studies indicate heterogeneous properties for this receptor. Although the basis for this heterogeneity is unknown, one possible explanation for these findings resides in the subunit composition of the receptor. Two 5‐HT3‐receptor subunits have been cloned: the 5‐HT3‐receptor subunit A (5‐HT3A) and the 5‐HT3‐receptor subunit B (5‐HT3B). Recombinant co‐expression of 5‐HT3A and 5‐HT3B subunits produces a functional heteromeric 5‐HT3A/3B receptor with pharmacological and electrophysiological properties different from those displayed by the 5‐HT3A homomeric receptor. In the present report, we used in situ hybridization histochemistry to demonstrate that the 5‐HT3B subunit is expressed in rat dorsal root ganglion (DRG) neurons. We determined with cellular resolution that 5‐HT3B subunit mRNA was expressed in 43.2 ± 2.8% of the total population of DRG neurons. By comparison, the 5‐HT3A subunit was more widely expressed, with 70.0 ± 2.8% of the total population of DRG neurons expressing this subunit. Further analyses showed that most of the neurons containing mRNA for the 5‐HT3B subunit (91.5 ± 3.4%) also expressed the 5‐HT3A subunit. In contrast, nearly half the population of neurons expressing 5‐HT3A subunit lacked (52.8 ± 5.9%) transcripts for the 5‐HT3B subunit. These results provide the first evidence indicating that the 5‐HT3B subunit of the 5‐HT3 receptor is expressed in DRG and suggest that sensory neurons have the capacity to synthesize at least two structurally different 5‐HT3 receptors: a heteromeric 5‐HT3A/3B receptor and a homomeric 5‐HT3A receptor. Consequently, 5‐HT3 receptors with different properties might be present in peripheral and central axons of the DRG. These findings open the possibility that distinct types of 5‐HT3 receptors may be involved in perception and/or processing of sensory information. J. Comp. Neurol. 438:163–172, 2001. Published 2001 Wiley‐Liss, Inc.
Diurnal and circadian variation of protein kinase C immunoreactivity in the rat retina
Authors: Gabriel Robert , Lesauter Joseph , Silver Rae , Garcia‐España Antonio , Witkovsky Paul
Published: August 2002

We studied the dependence of the expression of protein kinase C immunoreactivity (PKC‐IR) in the rat retina on the light:dark (LD) cycle and on circadian rhythmicity in complete darkness (DD). Two anti‐PKC alpha antibodies were employed: One, which we call PKCαβ recognized the hinge region; the other, here termed PKCα, recognized the regulatory region of the molecule. Western blots showed that both anti‐PKC antibodies stained an identical single band at approximately 80 kD. The retinal neurons showing PKC‐IR were rod bipolar cells and a variety of amacrine neurons. After 3 weeks on an LD cycle, PKCαβ‐IR in both rod bipolar and certain amacrine cells manifested a clear rhythm with a peak at zeitgeber time (ZT) of 06–10 hours and a minimum at ZT 18. No rhythm in total PKC‐IR was observed when using the PKCα antibody, but, at ZT 06–10 hours, rod bipolar axon terminals showed increased immunostaining. After 48 hours in DD, with either antibody, rod bipolar cells showed increased PKC‐IR. The PKCα antibody alone revealed that, after 48 hours, AII amacrine neurons, which lacked PKC‐IR in an LD cycle, manifested marked PKC‐IR, which became stronger after 72 hours. Light administered early in the dark period greatly increased PKCαβ‐IR in rod bipolar and some amacrine neurons. Our data indicate that light and darkness exert a strong regulatory influence on PKC synthesis, activation, and transport in retinal neurons. J. Comp. Neurol. 439:140–150, 2001. © 2001 Wiley‐Liss, Inc.
Physiological and anatomic evidence for functional subclasses of serotonergic raphe magnus cells
Authors: Gao Keming , Mason Peggy
Published: August 2002

Serotonergic cells in the medullary nucleus raphe magnus (RM) and adjacent nucleus reticularis magnocellularis (NRMC) project to the spinal cord where they are likely to modulate nociceptive transmission. Previous studies have suggested that these cells are physiologically and anatomically heterogeneous. In the present investigation, we examined whether subclasses of serotonergic RM and NRMC cells can be delineated based on their response to a visceral stimulus, and whether any such subclasses are morphologically distinct. Most RM and NRMC serotonergic cells tested (81 of 116) responded to retraction of the descending aorta into a polyethylene tube (the snare stimulus) with 57% of all cells tested excited and 13% inhibited. Responses of serotonergic cells to the snare outlasted the stimulus, were not reflective of evoked cardiovascular changes, and were observed in sino‐aortic deafferented rats, evidence that the snare stimulus does not influence serotonergic cell discharge through activation of baroreceptors. Because serotonergic cells responsive to the snare were also responsive to mechanical brushing within the retroperitoneum, the snare is likely to change serotonergic cell discharge by means of the activation of mechanosensitive visceral afferents. Intracellular labeling of physiologically characterized serotonergic RM and NRMC cells showed that cells that were responsive to the snare stimulus had simpler axonal collateralization patterns than cells that were unresponsive to the snare stimulus. This association between morphological and physiological properties provides additional evidence that subpopulations of serotonergic cells exist and serve varied physiological functions. J. Comp. Neurol. 439:426–439, 2001. © 2001 Wiley‐Liss, Inc.
“Type III” cells of rat taste buds: Immunohistochemical and ultrastructural studies of neuron‐specific enolase, protein gene product 9.5, and serotonin
Authors: Yee Cindy L. , Yang Ruibiao , Böttger Bärbel , Finger Thomas E. , Kinnamon John C.
Published: August 2002

Taste buds contain a variety of morphological and histochemical types of elongate cells. Serotonin, neuron‐specific enolase (NSE), ubiquitin carboxyl terminal hydrolase (PGP 9.5), and neural cell adhesion molecule (N‐CAM) all have been described as being present in the morphologically defined Type III taste cells in rats. In order to determine whether these substances coexist in a single cell, we undertook immunohistochemical and ultrastructural analysis of taste buds in rats. Double‐label studies show that PGP 9.5 and NSE always colocalize. In contrast, PGP 9.5 and serotonin seldom colocalize. Further, whereas the serotonin‐immunoreactive cells are always slender and elongate, the PGP 9.5/NSE population comprise two morphological types — one slender, the other broader and pyriform. Although gustducin‐immunoreactive taste cells appear similar in overall shape to the pyriform PGP 9.5/NSE population, gustducin never colocalizes with PGP 9.5 or NSE. The serotonin‐immunoreactive taste cells have an invaginated nucleus, synaptic contacts with nerve fibers, and taper apically to a single, large microvillus. These are all characteristics of Type III taste cells described previously in rabbits (Murray [1973] Ultrastructure of Sensory Organs I. Amsterdam: North Holland. p 1–81). PGP 9.5‐immunoreactive taste cells exhibit two morphological varieties. One type is similar to the serotonin‐immunoreactive population, containing an invaginated nucleus, synapses with nerve fibers, and a single large microvillus. The other type of PGP 9.5‐immunoreactive taste cell has a large round nucleus and the apical end of the cell tapers to a tuft of short microvilli, which are characteristics of Type II taste cells. Thus, in rats, some Type III cells accumulate serotonin but do not express PGP 9.5, whereas others express PGP 9.5 but do not accumulate amines. Similarly, Type II taste cells come in at least two varieties: those immunoreactive for gustducin and those immunoreactive for PGP 9.5. J. Comp. Neurol. 440:97–108, 2001. © 2001 Wiley‐Liss, Inc.
Estrogens modulate experimentally induced apoptosis of granule cells in the adult hippocampus
Authors: Liu Zhiping , Gastard Myriam , Verina Tatyana , Bora Susan , Mouton Peter R. , Koliatsos Vassilis E.
Published: August 2002

Estrogens are known to have broad effects on neuronal plasticity, but their specific role in neuronal cell death has not been determined. In the present study, we investigated the effects of β‐estradiol on an experimental model of apoptosis of granule cells of the dentate gyrus, i.e., apoptosis induced by intraventricular injection of the microtubule polymerization inhibitor colchicine. Cell death was characterized with multiple methods, including TUNEL and DNA electrophoresis. Nonrandom digestion of DNA was observed within 8–10 hours after colchicine injection, followed by condensation and fragmentation of granule cell nuclei and extensive anterograde degeneration of mossy fibers/terminals in 2 days. We compared the outcomes of the above‐described manipulation in ovariectomized or sham‐operated rats and animals treated daily with β‐estradiol or vehicle. Animals were lesioned with colchicine or vehicle 2 weeks after ovariectomy or sham operation. β‐Estradiol or vehicle was administered for 1 week prior to lesion and was continued for a further 2 weeks. Total numbers and densities of granule cells in different animal groups were counted by stereology in various anteroposterior levels of the hippocampus. Our results show that ovariectomy intensifies colchicine‐induced granule cell apoptosis, which is ameliorated by exogenous β‐estradiol. In doses that ameliorate the adverse effect of ovariectomy, exogenous β‐estradiol appears to have no effect of preventing granule cell death in animals with intact ovaries; i.e., an estrogen excess is not more neuroprotective than physiological levels of these hormones. Taken together, our results indicate that estrogen deprivation increases the vulnerability of hippocampal neurons to injury and may predispose to neurological diseases occurring after menopause. J. Comp. Neurol. 441:1–8, 2001. © 2001 Wiley‐Liss, Inc.
Analytic aspects of the Toda system: I. A Moser‐Trudinger inequality
Authors: Jost Jürgen , Wang Guofang
Published: August 2002

In this paper, we analyze solutions of the open Toda system and establish an optimal Moser‐Trudinger type inequality for this system. Let Σ be a closed surface with area 1 and K = (aij)N × N the Cartan matrix for SU(N + 1), i.e.,

We show that

has a lower bound in (H1(Σ))N if and only if

This inequality is optimal. As a direct consequence, if Mj < for 4π for j = 1, 2, …, N, ΦM has a minimizer u that satisfies

© 2001 John Wiley & Sons, Inc.
Hoare logic for Java in Isabelle/HOL
Authors: von Oheimb David
Published: August 2002

This article presents a Hoare‐style calculus for a substantial subset of Java Card, which we call Java$^{\ell ight}$. In particular, the language includes side‐effecting expressions, mutual recursion, dynamic method binding, full exception handling, and static class initialization.

The Hoare logic of partial correctness is proved not only sound (w.r.t. our operational semantics of Java$^{\ell ight}$, described in detail elsewhere) but even complete. It is the first logic for an object‐oriented language that is provably complete. The completeness proof uses a refinement of the Most General Formula approach. The proof of soundness gives new insights into the role of type safety. Further by‐products of this work are a new general methodology for handling side‐effecting expressions and their results, the discovery of the strongest possible rule of consequence, and a flexible Call rule for mutual recursion. We also give a small but non‐trivial application example.

All definitions and proofs have been done formally with the interactive theorem prover Isabelle/HOL. This guarantees not only rigorous definitions, but also gives maximal confidence in the results obtained. Copyright © 2001 John Wiley & Sons, Ltd.
The relationship of affect balance in family interactions to behaviour therapy outcome: a study with agoraphobic outpatients and their relatives
Authors: Renneberg Babette , Chambless Dianne L. , Fydrich Thomas , Goldstein Alan J.
Published: August 2002

The purpose of the study was to examine affect balance in dyads of patients and their relatives and its relationship to behaviour therapy outcome for agoraphobia and panic. Thirty‐five agoraphobic outpatients and their relatives participated in the study. On the basis of a 10‐min videotaped discussion between the patients and their partners, coded with Kategoriensystem für Partnerschaftliche Interaktion (KPI; Hahlweg et al., 1984), dyads were classified as affect balanced or unbalanced. Affect‐balanced dyads were those who demonstrated a preponderance of positive over negative and neutral behaviours for both people in the interaction. Compared to patients in unbalanced families, patients from balanced families were more likely to benefit from treatment on agoraphobic avoidance, although not on panic frequency, at both post‐test and a naturalistic 6‐month follow‐up. Copyright © 2002 John Wiley & Sons, Ltd.
Cognitive‐behavioural therapy for anxiety disordered children in a clinical setting: does additional cognitive parent training enhance treatment effectiveness?
Authors: Nauta Maaike H. , Scholing Agnes , Emmelkamp Paul M. G. , Minderaa Ruud B.
Published: August 2002

The effectiveness of a treatment protocol with individual CBT for children with anxiety disorders was evaluated in a clinical setting, i.e. an outpatient clinic for child and adolescent psychiatry. In addition, the surplus value of a cognitive parent‐training programme above the individual CBT was determined. Eighteen children with anxiety disorders were treated at an outpatient clinic for child and adolescent psychiatry. All children received 12 weekly sessions of individual cognitive behaviour therapy and all parents had two sessions in which the treatment method was explained. In addition, the families were randomly assigned to one of the following conditions: (1) no extra treatment, or (2) additional cognitive training for the parents, consisting of seven sessions, parallel to the individual treatment of the child. Results were studied at post‐treatment, at 3‐months follow‐up, and at 15 months follow‐up. Both child and parent reports showed significant decline of anxiety of the child, reaching the level of normal controls at 3 months follow‐up. According to parent reports, many children improved between the post‐test and the 3‐month follow‐up. Additional cognitive parent training did not add to the results of individual cognitive behaviour therapy for the children. At 15 months follow‐up, data for the total group revealed a general increase in anxiety reports in child reports but not in parent reports, compared with the three months follow‐up. Five children showed severe psychopathology, whereas the other 12 children reported no anxiety disorders and high levels of general functioning. Again, no differences between the treatment conditions were found. Copyright © 2001 John Wiley & Sons, Ltd.
State‐space equations of regular and strictly topologically degenerate linear lumped time‐invariant networks: the multiport method
Authors: Sommariva Antonino M.
Published: August 2002

A new complete approach to the multiport formulation of the state‐space equations of uniquely solvable regular or strictly topologically degenerate linear lumped time‐invariant networks is presented. It is based on a Gedankenexperiment during which the topological structure of the original network is manipulated in various ways. The final method requires one to calculate the describing matrices of three homogeneous multiports (i.e. a capacitive, an inductive and a resistive one), which are obtained from the network of interest in a very simple manner. As a by‐product, the equivalent partitioned network is also derived. As an example of application, the state‐space equations of a fourth‐order strictly topologically degenerate network are provided. Copyright © 2001 John Wiley & Sons. Ltd.
Cell analysis system based on compact disk technology
Authors: Tibbe Arjan G.J. , de Grooth Bart G. , Greve Jan , Rao Chandra , Dolan Gerald J. , Terstappen Leon W.M.M.
Published: August 2002

Background

A cell analysis system was developed to enumerate and differentiate magnetically aligned cells selected from whole blood. The cellular information extracted is similar to the readout of musical information from a compact disk (CD). Here we describe the optical design and data processing of the system. The performance of the system is demonstrated using fluorescent‐labeled cells and beads.

Materials and Methods

System performance was demonstrated with 6‐μm polystyrene beads labeled with magnetic nanoparticles and allophycocyanin (APC) and immunomagnetically aligned leukocytes, fluorescently labeled with Oxazine750 and CD4‐APC, CD8‐Cy5.5, and CD14‐APC/Cy7 in whole blood.

Results

The sensitivity of the system was demonstrated using APC‐labeled beads. With this system, beads containing 333 APC molecules could easily be resolved from the background. This level of sensitivity was not achievable with a commercial flow cytometer. A maximum of 20,000 immunomagnetically labeled cells could be aligned and analyzed in between 0.6 m of Ni lines, distributed over a surface area of 18 mm2 and extracted from a blood volume that depended on the height of the chamber. The utility of the system was demonstrated by performing a three‐color CD4‐CD8‐CD14 assay.

Conclusions

We built a cell analysis system based on immunomagnetic cell selection and alignment and analysis of fluorescent signals employing CD‐technology that is as good or better than current commercial analyzers. The cell analysis can be performed in whole blood or any other type of cell suspension without extensive sample preparation. Cytometry 47:173–182, 2002. © 2002 Wiley‐Liss, Inc.
Discrimination of DNA and RNA in cells by a vital fluorescent probe: Lifetime imaging of SYTO13 in healthy and apoptotic cells
Authors: van Zandvoort Marc A.M.J. , de Grauw Cees J. , Gerritsen Hans C. , Broers Jos L.V. , oude Egbrink Mirjam G.A. , Ramaekers Frans C.S. , Slaaf Dick W.
Published: August 2002

Background

Of the few vital DNA and RNA probes, the SYTO dyes are the most specific for nucleic acids. However, they show no spectral contrast upon DNA or RNA binding. We show that fluorescence lifetime imaging using two‐photon excitation of SYTO13 allows differential and simultaneous imaging of DNA and RNA in living cells, as well as sequential and repetitive assessment of staining patterns.

Methods

Two‐photon imaging of SYTO13 is combined with lifetime contrast, using time‐gated detection. We focus on distinguishing DNA and RNA in healthy and apoptotic Chinese hamster ovary cells.

Results

In healthy cells, SYTO13 has a fluorescence lifetime of 3.4 ± 0.2 ns when associated with nuclear DNA. Bound to RNA, its lifetime is 4.1 ± 0.1 ns. After induction of apoptosis, clusters of SYTO13 with fluorescence lifetime of 3.4 ± 0.2 ns become apparent in the cytoplasm. They are identified as mitochondrial DNA on the basis of colocalization experiments with the DNA‐specific dye, DRAQ5, and the mitochondrial‐specific dye, CMXRos. Upon progression of apoptosis, the lifetime of SYTO13 attached to DNA shortens significantly, which is indicative of changes in the molecular environment of the dye.

Conclusions

We have characterized SYTO13 as a vital lifetime probe, allowing repetitive and differential imaging of DNA and RNA. Cytometry 47:226–235, 2002. © 2002 Wiley‐Liss, Inc.
Heterogeneity of freshly isolated human tonsil dendritic cells demonstrated by intracellular markers, phagocytosis, and membrane dye transfer
Authors: Stent Gianna , Reece Jeanette C. , Baylis Dean C. , Ivinson Karen , Paukovics Geza , Thomson Megan , Cameron Paul U.
Published: August 2002

Background

Heterogeneity within human dendritic cells (DCs) has been described but its functional relationships to cells of macrophage lineage and its role in human immunodeficiency virus (HIV) infection in vivo remain unclear.

Methods

Tonsil macrophages and DCs were isolated from low‐density cells by negative selection and DCs were sorted into myeloid and plasmacytoid populations using antibodies to CD11c or CD123. Phagocytosis of latex beads and uptake of dye‐labeled target cells were compared by flow cytometry and CD68 and S‐100 by immunofluorescence on cytospins of sorted cells.

Results

Bead uptake and membrane dye transfer were found in both blood and tonsil CD11c+ DCs and in CD14+ cells particularly from blood monocytes. CD11c‐ DCs were poorly phagocytic but took up fluorescent dye from intact, necrotic or apoptotic cells. Tonsil DCs and macrophages expressed both CD68 and S‐100 but CD11c‐ DCs expressed CD68 only.

Conclusions

Freshly isolated CD11c+ tonsil DCs are similar to CD14+ macrophages in phagocytic function but the poorly phagocytic CD11c‐ DCs can also take up membrane from target cells. The intracellular markers commonly used to identify DCs and macrophages in situ do not identify accurately the CD11c‐ DC subset nor do they distinguish tonsil macrophages from DCs. Cytometry 48:167–176, 2002. © 2002 Wiley‐Liss, Inc.
Neuropathic pain and the electrophysiology and pharmacology of nerve injury
Authors: Smith Peter A. , Stebbing Martin J. , Moran Timothy D. , Tarkkila Pekka , Abdulla Fuad A.
Published: August 2002

Nociceptive pain serves the useful purpose of alerting the body to potential or actual tissue damage. By contrast, neuropathic pain that results from injury or damage to the nervous system persists long after all signs of the original injury have disappeared. Neuropathic pain presents a significant clinical problem as it responds poorly to classical analgesics such as non‐steroidal anti‐inflammatory drugs (NSAIDs) and to opioids; there is also no single, uniformly well‐tolerated drug that is reliably helpful. Treatment currently involves the use of anticonvulsant and/or antidepressant drugs. Electrophysiological experiments on dorsal root ganglion and spinal cord neurons of nerve‐injured experimental animals are yielding new information on the pathophysiology of neuropathic pain. Analysis of actions of various neuropeptides and neurotransmitters in these models has helped to explain the poor efficacy of opioids and suggests new therapeutic approaches to the management of neuropathic pain. Drugs that stimulate α2‐c‐adrenoceptors or that mimic the actions of neuropeptide Y, galanin, or the opioid‐like peptide, nociceptin, may be of use in this regard. Drug Dev. Res. 54:140–153, 2002. © 2002 Wiley‐Liss, Inc.
Persistence of nicotinic agonist RJR 2403‐induced working memory improvement in rats
Authors: Levin Edward D. , Christopher N. Channelle
Published: August 2002

Nicotinic systems are involved in the neural basis of memory function and nicotinic agonists have shown promise for the treatment of cognitive dysfunction. Both acute and chronic nicotinic treatment has been shown to improve memory performance. There is some evidence for the persistence of nicotine‐induced memory improvements lasting longer than the pharmacokinetic presence of nicotine. Novel nicotinic agonists may produce the beneficial effects of nicotine on cognitive function with fewer side effects. RJR 2403 (metanicotine or (E)‐N‐methyl‐4‐(3‐pyridinyl)‐3‐butene‐1‐amine) a preferential α4β2 nicotinic agonist has been shown in previous studies to improve memory function. The present study examined the persistence of oral RJR 2403 on memory performance in young adult Sprague‐Dawley rats (3–5 months old) 30 min, 1 h, and 6 h post‐administration (PO). In the first experiment, the typical inverted U‐shaped dose response curve for cognitive enhancing drugs was seen with RJR 2403 30 min after administration. The lower dose of 1 mg/kg (3.6 μmol/kg), but not higher doses of 3 or 10 mg/kg (10.8 or 36 μmol/kg), of RJR 2403 significantly (P < 0.05) improved working memory on the radial‐arm maze relative to placebo‐treated controls. In the second experiment, young adult rats were administered RJR 2403 (0, 0.3, or 1.0 mg/kg) and tested 1 h or 6 h following administration in separate treatment groups. The 0.3 mg/kg RJR 2403 dose (1.08 μmol/kg) caused a significant (P < 0.05) memory improvement 1 h after oral administration. Interestingly, both the 0.3 and 1.0 mg/kg RJR 2403 doses (1.08 and 3.6 μmol/kg) significantly (P < 0.05) improved memory six hours after administration. These data demonstrate the efficacy of oral RJR 2403 in improving cognitive performance and the long duration of action of RJR 2403 in young adult rats. In contrast, no significant memory improvement was seen in aged rats aged (24–26 months old) after RJR 2403 administration. The inability of RJR 2403 to enhanced cognitive functions in aged rats might be related to the decrease in the number of α4β2 nicotinic receptors, which occurs with age. A similar decreased responsiveness in aged rats has been seen with nicotine. The persistence of action of RJR 2403 provides additional promise for its potential as a treatment for cognitive dysfunction. However, the decreased responsiveness in subjects in populations with decreased nicotinic receptor number may limit the effectiveness of nicotinic therapies. Drug Dev. Res. 55:97–103, 2002. © 2002 Wiley‐Liss, Inc.
Stretch‐activated channels in the heart: Their role in arrhythmias and potential as antiarrhythmic drug targets
Authors: Saint David A.
Published: August 2002

Stretching the myocardium can have profound effects on its electrical activity, a process called mechanoelectric feedback. Mechanoelectric feedback can be powerful enough to trigger arrhythmias, and it is likely to be the underlying trigger for some clinically intractable arrhythmias. It seems likely that the response to stretch involves at least two types of mechanosensitive ion channels, a nonspecific cation channel, and a potassium channel (or channels). Currents carried by nonspecific cation channels have been well characterised in cardiac tissue, although the gene has not yet been identified. Very recently, a peptide toxin isolated from Grammastola spatulata has been shown to block this channel and to be an effective antiarrhythmic agent against stretch‐induced atrial arrhythmias. Stretch‐activated potassium currents and single channels have also been recorded in cardiac cells; in the case of these channels, the genes coding for them have recently been cloned and shown to be members of a new potassium channel gene family, the “tandem pore” channels. Although some nonspecific activators and blockers of these channels exist, there is at present no specific blocker, which limits investigation of the role of these channels in arrhythmogenesis. The potential for the development of blockers of both types of stretch‐activated channels as therapeutic agents is discussed. Drug Dev. Res. 55:53–58, 2002. © 2002 Wiley‐Liss, Inc.
Cardiovascular responses to pacifier experience and feeding in newborn infants
Authors: Cohen Morris , Brown David R. , Myers Michael M.
Published: August 2002

This study examined the effects of sucking on a pacifier immediately before feeding on cardiovascular responses to feeding. Twenty‐eight bottle‐feeding infants were studied at 12 to 40 hr of age. Blood pressure and heart rate measurements were made during three periods: Period 1—while infants rested in their cribs before a regularly scheduled feeding, Period 2—while being held by the feeder immediately before feeding, and Period 3—during the first 3 to 5 min of feeding. Half of the infants were given a pacifier during Period 2. Blood pressures and heart rates increased across the periods; however, increases in systolic blood pressure during feeding were reduced for the group of infants given a pacifier prior to feeding. We suggest that cardiovascular responses to feeding in infants are comprised of multiple elements. Most of the systolic blood pressure responses to feeding are elicited by sucking whereas the heart rate response is dependent on both sucking and nutrient intake. © 2001 John Wiley & Sons, Inc. Dev Psychobiol 39: 34–39, 2001
Wherefore heart thou? Embryonic origins of cardiogenic mesoderm
Authors: Yutzey Katherine E. , Kirby Margaret L.
Published: August 2002

The developing heart in avian embryos has been examined extensively over the past several decades using classic embryologic and, more recently, molecular and genetic approaches. Still, conflicting reports arise as to the location and regulation of early heart progenitors in the embryo. In addition, a new source of cardiomyocytes has been identified recently that contributes to the outflow tract after the heart initially forms. The focus of this review is the examination of the tissue interactions, signaling molecules, and gene regulatory mechanisms that, together, control heart formation from primary and secondary heart forming fields of the embryo. Early studies of the induction and regulation of the secondary heart field indicate that at least some of the events of primary cardiomyogenesis are recapitulated when the conotruncal myocardium is recruited into the heart. The consideration of classic embryologic studies of the heart forming fields in terms of modern molecular and genetic tools provides reinforcing evidence for the location of cardiac progenitors in the embryo. The accurate definition of early cardiac regulatory events provides a necessary foundation for the generation of new therapeutic sources of cardiomyocytes. © 2002 Wiley‐Liss, Inc.
Taste bud development in the zebrafish, Danio rerio
Authors: Hansen Anne , Reutter Klaus , Zeiske Eckart
Published: August 2002

Taste buds are chemosensory endorgans consisting of modified epithelial cells. Fish and other vertebrates use their taste bud cells to sample potential food, either selecting or rejecting substances according to their edibility. The adult gustatory system in fish has been studied thoroughly, including regeneration experiments. Taste buds occur in the epithelia of the lips, the mouth cavity, the oropharyngeal cavity, and also in the skin of the barbels, the head, and sometimes even all over the body surface. Despite its importance for feeding, little is known about the ontogeny of the fish taste system. We examined the development of taste buds in the zebrafish on the light microscopical and the scanning and transmission electron microscopical levels. Taste buds develop later than the olfactory organ and the solitary chemosensory cells, two other chemosensory systems in aquatic vertebrates. The first few taste bud primordia are visible within the epithelia of lips and gill arches 3 to 4 days after fertilization, and the first few taste buds with open receptor areas appear on the lips and simultaneously on the gill arches 4–5 days after fertilization, which coincides with the onset of feeding. Taste buds in the mouth cavity, on the head, and on the barbels are formed later in development. As seen in other fish, zebrafish taste buds contain elongate dark and light cells, termed according to their electron density. Dark cells with a cell apex of many short microvilli appear first, followed by the light cells with one large microvillus. In addition, the zebrafish has a third fusiform cell type, which appears last. This cell type is low in electron density and has a brush‐like apical ending with several small microvilli. This cell type has not been described previously. Furthermore, in zebrafish, the ontogenetic processes of taste bud formation differ from regenerative processes described in the literature. © 2002 Wiley‐Liss, Inc.
Postembryonic expression of Caenorhabditis elegans mab‐21 and its requirement in sensory ray differentiation
Authors: Ho Siu H. , So Gloria M.K. , Chow King L.
Published: August 2002

The male tail sensory rays in Caenorhabditis elegans are complex copulatory structures, the normal patterning of which requires a number of regulatory genes. Among them, mab‐21 specifies the identity of sensory ray 6. By using green fluorescent protein reporters, we identify multiple cis‐acting elements that control the developmental expression of mab‐21. Traced with a functional mab‐21:gfp gene driven by authentic regulatory sequences, mab‐21 expression could be detected in hypodermal, neuronal, muscle, and ray cells. We showed here that the expression of mab‐21 in the hypodermis and neuronal cells was dispensable for its function in ray 6. In contrast, its expression in the ray 6 structural cell and neurons as conferred by its 3′ enhancer was crucial for determining the correct ray 6 identity. © 2001 Wiley‐Liss, Inc.
Day hospitalization programs for eating disorders: A systematic review of the literature
Authors: Zipfel Stephan , Reas Deborah L. , Thornton Chris , Olmsted Marion P. , Williamson Donald A. , Gerlinghoff Monika , Herzog Wolfgang , Beumont Peter J.
Published: August 2002

Objective

Partial day hospitalization programs for the treatment of eating disorders are increasingly being developed worldwide.

Method

First, a systematic database search of day hospitalization programs for eating disorders, published in either English or German, was conducted. Programs that provided sufficient information on their program structure were summarized and compared across various dimensions. Second, the responsible program directors were contacted to provide additional information regarding outcome data, current trends, challenges, and future directions of their programs. Third, outcome data from day programs presented at international conferences were included to expand the base of the review.

Discussion

Although the programs from different countries and health care environments varied in terms of their purpose and operated within very different health care systems, many similarities were found to exist, including the use of a multidisciplinary staff and reliance on group treatment as the primary means of therapy. Marked differences were noted in terms of inclusion criteria and intensity of care. © 2002 by Wiley Periodicals, Inc. Int J Eat Disord 31: 105–117, 2002; DOI. 10.1002/eat.10009
Development of body image, eating disturbance, and general psychological functioning in adolescent females: a replication using covariance structure modeling in an Australian sample
Authors: van den Berg Patricia , Wertheim Eleanor H. , Thompson J. Kevin , Paxton Susan J.
Published: August 2002

Objective

The current study is a follow up to a covariance structure modeling (CSM) investigation of predictors of body image, eating dysfunction, and general psychological functioning in U.S. adolescent females.

Method

Four hundred and seventy Australian girls (13–17 years old) completed measures of teasing history, body dissatisfaction, global psychological functioning, dietary restraint, and bulimic behaviors. They were also measured in order to compute body mass index (BMI).CSM techniques were used to analyze the relationships between these latent variables. Two a priori models were tested, one was selected, and post‐hoc model‐fitting was undertaken.

Results

The final model indicated that BMI directly influenced teasing, which directly influenced body dissatisfaction. Body dissatisfaction influenced both global psychological functioning and restriction, whereas restriction had a direct influence on bulimia. Global psychological functioning also had a direct influence on bulimia.

Discussion

The results support previous research indicating that teasing and body dissatisfaction are risk factors for the development of eating disturbances. © 2002 by Wiley Periodicals, Inc. Int J Eat Disord 32: 46–51, 2002.
Pathways mediating sexual abuse and eating disturbance in children
Authors: Wonderlich Stephen , Crosby Ross , Mitchell James , Thompson Kevin , Redlin Jennifer , Demuth Gail , Smyth Joshua
Published: August 2002

Objective

To examine the relationship between childhood maltreatment and eating disorders in a sample of children.

Method

Twenty 10–15‐year‐old female children who were receiving treatment following reported childhood sexual abuse and 20 age‐matched controls were compared on a series of measures assessing eating disorder behaviors, body image concerns, substance use, mood, impulsive behavior, and self‐concept.

Results

Sexually abused children reported higher levels of eating disorder behaviors, impulsive behaviors, and drug abuse than controls. Furthermore, behavioral impulsivity provided the strongest mediational effect between a history of childhood sexual abuse and purging and restrictive dieting behavior. Drug use proved to be a significant secondary mediator of the childhood sexual abuse eating disorder behavior association.

Discussion

These data support the hypothesis that childhood sexual abuse is related to disordered eating in children, and extend similar findings that have been previously reported with adults. Behavioral impulsivity and drug use appear to be significant mechanisms that influence eating disorder behavior following childhood sexual abuse. © 2001 by John Wiley & Sons, Inc. Int J Eat Disord 29: 270–279, 2001.