Viral interleukin-6 encoded by rhesus macaque rhadinovirus is associated with lymphoproliferative disorder (LPD).

BACKGROUND: Rhesus macaques (RM) co-infected with simian immunodeficiency virus (SIV) and rhesus macaque rhadinovirus (RRV) develop abnormal cellular proliferations characterized as extra-nodal lymphoma and retroperitoneal fibromatosis (RF). RRV encodes a viral interleukin-6 (vIL-6), much like Kaposi's sarcoma-associated herpesvirus, and involvement of the viral cytokine was examined in proliferative lesions. METHODS: Formalin fixed tissue from RM co-infected with SIV and RRV were analyzed for RRV genomes by in situ hybridization and RRV vIL-6 expression by immunofluorescence analysis. RESULTS: In situ hybridization analysis indicated that RRV is present in both types of lesions. Immunofluorescence analysis of different lymphomas and RF revealed positive staining for vIL-6. Similarly to KS, RF lesion is positive for vimentin, CD117 (c-kit), and smooth muscle actin (SMA) and contains T cell, B cell and monocytes/macrophage infiltrates. CONCLUSIONS: Our data support the idea that vIL-6 may be critical to the development and progression of lymphoproliferative disorder in RRV/SIV-infected RM.

A multicenter comparison study between the Endosafe PTS rapid-release testing system and traditional methods for detecting endotoxin in cell-therapy products.

BACKGROUND: Rapid-release testing reduces the waiting period for administration of time-sensitive cell-therapy products. Current assay systems are labor intensive and time consuming. The Endosafe portable test system (PTS) is a chromogenic Limulus amebocyte lysate (LAL) portable endotoxin detection system that provides quantitative results in approximately 15 min. To evaluate Endosafe performance with cell-therapy products, side-by-side testing of traditional LAL systems and the Endosafe system was conducted at the Production Assistance for Cellular Therapies (PACT) facilities and the National Institutes of Health's Department of Transfusion Medicine, USA. METHODS: Charles River Laboratories provided each center with a PTS reader and two commercially prepared lyophilized reference standard endotoxin (RSE) vials. All samples tested with the Endosafe system used 0.05-5.0 endotoxin unit/mL (EU/mL) sensitivity cartridges provided by Charles River. Each vial was reconstituted with LAL water and tested in triplicate using the Endosafe and in-house LAL methods. Subsequently, each center tested the endotoxin content of standard dilutions of cell-therapy products, thus creating paired test results for each sample. Additionally, fabricated endotoxin-positive samples containing varying concentrations of endotoxin were prepared and shipped to all centers to perform blinded testing. RESULTS: Valid paired results, based on each center's LAL method and the Endosafe system criteria, were analyzed. Endotoxin detection between paired results was equivalent in most cases. DISCUSSION: The Endosafe system provided reliable results with products typically produced in cell-therapy manufacturing facilities, and would be an appropriate test on which to base the release of time-sensitive cell-therapy products.

Influence of dietary fats on Ecstasy-induced hyperthermia.

BACKGROUND AND PURPOSE: Studies were designed to examine the effects of dietary fats on metabolic effects of 3,4-methylenedioxymethamphetamine (MDMA, Ecstasy). These effects included hyperthermia, expression of uncoupling protein (UCP1 and 3) in brown adipose tissue or skeletal muscle and plasma free fatty acid (FFA) levels. EXPERIMENTAL APPROACH: Male Sprague-Dawley rats were fed either a high-fat diet (HFD, 60% kcal) or a lower fat isocaloric controlled diet (LFD, 10% kcal) for 28 days before MDMA challenge. KEY RESULTS: No significant differences were observed between LFD and HFD groups in terms of body weight, plasma thyroxine (T4) levels and expression of brown fat UCP1 or skeletal muscle UCP3 protein. HFD significantly raised levels of circulating FFA and potentiated the thermogenesis induced by MDMA (10 mg kg(-1), s.c.), compared to the effects of the LFD. Moreover, 30 and 60 min after MDMA administration, plasma FFA levels decreased in HFD animals, but were markedly elevated in the LFD group. CONCLUSIONS AND IMPLICATIONS: These results indicate that high-fat feeding regulates MDMA-induced thermogenesis by augmenting the activation of UCP rather than its expression.

Bombardment-induced tunable superlattices in the growth of Au-Ni films.

Highly ordered superlattices are typically created through the sequential deposition of two different materials. Here, we report our experimental observation of spontaneous formation of superlattices in coevaporation of Au and Ni under energetic ion bombardment. The superlattice periodicities are on the order of a few nanometers and can be adjusted through the energy and flux of ion beams. Such a self-organization process is a consequence of the bombardment-induced segregation and uphill diffusion within the advancing nanoscale subsurface zone in the film growth. Our observations suggest that ion beams can be employed to make tunable natural superlattices in the deposition of phase-separated systems with strong bombardment-induced segregation.

Unilateral infusion of a dopamine transporter antisense into the substantia nigra protects against MDMA-induced serotonergic deficits in the ipsilateral striatum.

The present study was designed to elucidate the consequences of antisense oligonucleotide-mediated knockdown of striatal dopamine reuptake transporters on 3,4-methylenedioxymethamphetamine (MDMA)-induced neurotoxicity. Antisense oligonucleotide complementary to the mRNA translational start site of the rat dopamine transporter was delivered by constant (7 days) intranigral infusion with an osmotic minipump. Delivery of the antisense oligonucleotide by this method resulted in a 70% reduction in the density of the dopamine transporter in the ipsilateral striatum, as measured by [(3)H]mazindol binding. The effect of this transporter knockdown on MDMA-induced serotonergic neurotoxicity was then examined. MDMA (2x20 mg/kg, s.c., given 12 h apart) administered to control rats produced hyperthermia following the first dose and led to a 45-50% reduction in striatal serotonin, 5-hydroxyindoleacetic acid, and serotonin reuptake transporter density 1 week after the second dose. Conversely, in antisense-, but not missense-treated rats, a significant attenuation of MDMA-induced neurotoxicity was observed only in the ipsilateral striatum. The hyperthermic response elicited by MDMA was not altered by prior administration of antisense. In vivo microdialysis revealed that the antisense treatment attenuated MDMA-induced dopamine release in the ipsilateral striatum. These results suggest that the dopamine transporter plays an essential role in the neurodegeneration induced by MDMA, and provides additional support for the hypothesis that extracellular dopamine is involved in the neurotoxic process, at least in the striatum.

Temperature-dependent activation of recombinant rat vanilloid VR1 receptors expressed in HEK293 cells by capsaicin and anandamide.

Capsaicin activates vanilloid (VR1) receptors found on sensory neurons. These ligand-gated ion channels are also sensitive to low pH, elevated temperature and the endocannabinoid, anandamide. In this study, we have measured capsaicin- and anandamide-induced elevations in intracellular calcium concentrations ([Ca(2+)](i)) in fura-2 loaded HEK293 cells stably expressing the rat VR1 receptor at 22, 37 and 50 degrees C. Both capsaicin and anandamide produced a concentration-dependent elevation in [Ca(2+)](i) at all temperatures. pEC(50) values were 7.74 and 5.69 at 22 degrees C and 6.90 and 5.15 at 37 degrees C for capsaicin and anandamide, respectively. At 50 degrees C, the pEC(50) value for capsaicin was 6.36 but the response to anandamide did not saturate. Responses to both agonists were sensitive to ruthenium red and capsazepine at all temperatures. This temperature-dependent reduction in potency may result from desensitization.

The pharmacodynamic characterization of an antisense oligonucleotide against monoamine oxidase-B (MAO-B) in rat brain striatal tissue.

1. The aim of our work was to pharmacodynamically characterize an antisense oligonucleotide sequence (5'-GCC AAA CTT TTG CAT GAC-3') against MAO-B, using qualitative and quantitative analyses as assessment measures. 2. Qualitative analysis using histochemical staining revealed that intracerebroventricular (ICV) administered antisense (100 picomoles twice daily x 3.5 days) eliminated all visibly detectable histochemical staining for MAO-B throughout the striatum 1, 12, and 24 h after the last antisense treatment. 3. Qualitative analysis using RT-PCR of the time course of MAO-B mRNA expression in the rat striatum following ICV administration of the antisense sequence showed that 12-24 h after the last administration there was a dramatic reduction in MAO-B mRNA expression in the striatum. The reverse and scrambled sequences generated no change in MAO-B mRNA at 1 or 24 h after the last treatment. 4. Quantitative analysis using the MAO-B selective substrate 4-dimethylamino-phenethylamine (DMAPEA) showed that the antisense sequence reduced MAO-B activity by more than 40%, which was comparable to a single 2 mg/kg, ip dose of L-deprenyl. 5. Quantitative analysis of neurotransmitter levels 24 h after the last treatment suggested that the antisense sequence did not produce any significant changes in neurotransmitter levels. 6. Potential mechanisms for enhancing the antisense response and the speculated potential of an antisense against MAO-B for studying neurotoxicity, Parkinson's disease, and the aging process are also discussed.

The Zebrafish Information Network (ZFIN): a resource for genetic, genomic and developmental research.

The Zebrafish Information Network, ZFIN, is a WWW community resource of zebrafish genetic, genomic and developmental research information (http://zfin.org). ZFIN provides an anatomical atlas and dictionary, developmental staging criteria, research methods, pathology information and a link to the ZFIN relational database (http://zfin. org/ZFIN/). The database, built on a relational, object-oriented model, provides integrated information about mutants, genes, genetic markers, mapping panels, publications and contact information for the zebrafish research community. The database is populated with curated published data, user submitted data and large dataset uploads. A broad range of data types including text, images, graphical representations and genetic maps supports the data. ZFIN incorporates links to other genomic resources that provide sequence and ortholog data. Zebrafish nomenclature guidelines and an automated registration mechanism for new names are provided. Extensive usability testing has resulted in an easy to learn and use forms interface with complex searching capabilities.

Self-determination and empowerment: a feminist standpoint analysis of talk about disability.

In this paper we offer a feminist analysis of talk about self-determination and empowerment in the context of disability, focusing on the case of developmental disabilities. We find strains of the same patterns feminist epistemologists have argued shape the organization of formal knowledge from the standpoint of the privileged. At the extreme, people with developmental disabilities appear as objects without selves, outside of the context of interpersonal and social structural relationships that constrain who they can be by defining them as other, often in multiple and interacting ways. Empowerment, from the dominant standpoint, becomes an abstract attribute or condition; something a person has or does not have. Taking the standpoint of women and other marginalized people offers a view of self-determination as a person's development of his or her self. Empowerment becomes a potential characteristic of a social relationship, one that facilitates the development of someone's self. The most empowering relationships are mutual, recognizing and building on the diverse contributions and needs of participants in ways that seek to minimize inequalities over time. The reason some of us are self-determining is that we are in interpersonal and social structural relationships that empower us. To construct interpersonal and social structural relationships that empower people with developmental disabilities requires challenging the way dominant conceptualizations of independence and productivity also express the standpoint of the privileged. The standpoint of women allows all of us to talk more of how we connect with and facilitate one another's developing selves within communities.

A risky business.

Just when you get comfortable with life--you've got everything under control and you know what you're doing--along comes someone with a new idea with a change attached to it. From the European Courts, through the Government to the Trust Executive and then the Directorates, change seems to be a constant part of life. Julie Sprague is a theatre nurse who saw an opportunity to make a worthwhile change in her area of practice. This is the acceptable face of change, at clinical practice level, done with involvement and co-operation of colleagues. This article describes that change process together with the theories and rationale which support it.

Cast syndrome: the superior mesenteric artery syndrome.

Cast syndrome, clinically known as superior mesenteric artery syndrome (SMAS), is gastric dilatation with partial or complete obstruction of the duodenum. Although rare, it is most frequently seen in orthopaedic patients who have had spinal surgery or who are in hip spica or body casts. Obstruction occurs when there is compression of the duodenum between the superior mesenteric artery anteriorly and the aorta and spinal column posteriorly. Obstruction can occur within days of surgery or casting or may not develop for several weeks. Treatment for SMAS varies from conservative nonoperative to operative procedures. Complications can be severe if symptoms are not quickly recognized and treatment instituted in a timely manner.

An integrated hypothesis for the serotonergic axonal loss induced by 3,4-methylenedioxymethamphetamine.

Administration of the street drug 3,4-methylenedioxymethamphetamine (MDMA) to various experimental animals has been shown in several laboratories to induce selective damage to serotonergic axons and axon terminals. This review examines the current available evidence supporting the development of serotonin (5-HT) neurotoxicity in animals and humans. There are a plethora of hypotheses that attempt to explain the mechanisms involved in the development of this serotonergic neurotoxicity. An integrated hypothesis incorporating most of the speculated neurotransmitters theorized to be involved in the process is proposed. This hypothesis states that MDMA induces the following sequence of events resulting in the serotonergic neurotoxicity: 1. MDMA induces an acute release of 5-HT and dopamine (DA). 2. This acute release is followed by depletion of intraneuronal 5-HT stores. 3. The initially released 5-HT activates post-synaptic 5-HT2A/2C receptors located on GABA interneurons resulting in a decrease in GABAegic transmission and increased DA release and synthesis. 4. The excessive DA released then may be transported into the depleted 5-HT terminal. 5. The DA is then deaminated by monoamine oxidase B (MAO-B) located within the 5-HT terminal. This results in free-radical formation and the selective degeneration of the serotonergic axons and axon terminals. While there is no clear evidence that human users of the drug are suffering a similar neurotoxicity, data are presented suggesting that there remains cause for concern.

Interleukin-12 p40 m-RNA expression in human kidney allograft biopsies.

Interleukin-12 (IL-12) is a heterodimeric cytokine implicated in the early differentiation of naive T-lymphocytes into the Th1 subset. IL-12 is important for induction of the cellular immune response against viruses, intracellular parasites and neoplasms. Its role in alloresponsiveness has not been fully elucidated. Preliminary data in the literature point toward the prevalence of Th1 lymphocytes in processes of allograft rejection. In attempt to further investigate the expression of this cytokine during episodes of cellular rejection of renal allografts, we searched for IL-12 message in human kidney allograft biopsies using the reverse transcriptase-polymerase chain reaction technique. Twenty-three allograft core biopsies from 19 patients were obtained percutaneously for clinical indications in 18 cases, and as part of an investigational protocol in five cases. A portion of the tissue was used for RNA extraction using the guanidium-thiocyanide phenol-chloroform method. Histology was performed on the remaining core material. Ten mg of total RNA were used for reverse transcription. PCR of the c-DNAs was done for 40 cycles using primers for the p40 subunit of IL-12 and GAPDH which was used as a control. PCR products were photographed after electrophoresis, transferred to a nylon membrane and hybridized with a radiolabelled cloned human IL-12 p40 1 kb c-DNA fragment. Autoradiographies were developed after 20-min exposure. All samples were run in triplicate. IL-12 p40 m-RNA was expressed in all 17 biopsies showing acute cellular rejection as well as in all three biopsies showing focal interstitial fibrosis. No message was found in the presence of normal allograft histology. This is the first in vivo report of IL-12 p40 subunit m-RNA expression during renal allograft rejection in humans. The role of this Th1 cytokine in the alloresponse deserves further investigation.

The effect of noncontingent sensory reinforcement, contingent sensory reinforcement, and response interruption on stereotypical and self-injurious behavior.

Three analyses were conducted to assess the effects of different consequent stimuli on the rate of stereotypical and self-injurious behavior performed by two individuals with severe developmental disabilities and dual sensory impairments. An analogue functional analysis documented an undifferentiated pattern of problem behavior across all conditions for Participant 1. Data for Participant 2 indicated an undifferentiated pattern with lower frequencies in the demand condition. Stimuli chosen to compete with the type of sensory stimulation produced by the stereotypy and self-injurious behavior were presented noncontingently during play conditions. Noncontingent presentation of the specially selected stimuli resulted in reductions in stereotypy and self-injurious behavior. Finally, contingent presentation of the same stimuli with and without response interruption was assessed in a demand context. Contingent presentation of the specially selected stimuli plus response interruption resulted in more suppression than contingent sensory stimulus presentation alone. Results are discussed as to competing and concurrent schedules of reinforcement.

Studies on the mechanism of p-chloroamphetamine neurotoxicity.

Studies were conducted to investigate the sensitivity of p-chloroamphetamine (PCA)-induced neurochemical changes to various pharmacological manipulations known to block the neurochemical effects of 3,4-methylenedioxymethamphetamine (MDMA). The monoamine oxidase-B (MAO-B) inhibitor L-deprenyl (2 mg/kg) given 4 hr before a nonneurotoxic dose of PCA (2 mg/kg) was shown not to alter the amount of [3H]paroxetine bound to serotonin (5-HT) uptake sites 7 days after treatment. L-Deprenyl 4 hr before a neurotoxic dose of PCA (10 mg/kg) did not change the acute hyperthermia. Further, neither L-deprenyl nor another selective MAO-B inhibitor, MDL-72,974 (1.25 mg/kg), given 30 min before or daily for 4 days before a single dose of PCA attenuated or potentiated the decrease in the number of [3H]paroxetine binding sites measured 7 days after PCA treatment. The combination of the MAO-A inhibitor clorgyline (2.5 mg/kg) or a nonspecific dose of L-deprenyl (10 mg/kg) with the selective 5-HT releasing agent 5,6-methylenedioxy-2-aminoindan did not lead to changes in the levels of 5-HT, 5-hydroxyindoleacetic acid or dopamine 7 days after treatment. Finally, the 5-HT2A receptor antagonist MDL-11,939 (5 mg/kg) did not protect against the neurotoxicity of PCA. By comparing the present work with previous studies of MDMA, these results can be interpreted to suggest that the mechanism of the neurotoxicity induced by PCA is not identical to that induced by MDMA. The relationship of these results to the neurotoxicity induced by MDMA is also discussed.

Building blocks. Roundtable discussion.

Former Hospitals & Health Networks executive editor Mark Hagland and Health Facilities Management editor Kurt Luchs and associate editor Catherine Quayle recently hosted a roundtable discussion on design and construction issues in health care. They wanted to know what health care senior executives and experts in the design and construction field thought about today's most pressing issues and biggest trends. Hagland and Luchs assembled a group or experts and executives in Chicago and set the tape rolling. Here are excerpts from the discussion.

Orientation discrimination in the cat: its cortical locus II. Extrastriate cortical areas.

Luminance-defined edges or bars are among the basic units of visual analysis: a "primitive" component of perception. We have utilized this stimulus in a psychophysical study of bar orientation discrimination in the cat before and after selective lesions in visual cortical areas. The cortices have been divided on the basis of their connectivity into three tiers. Tier I refers to areas 17 and 18, tier II includes areas that receive directly from tier I, and tier III includes those areas that receive directly from tier II. Previous studies (Vandenbussche et al. [1991] J. Comp. Neurol. 305:632-658) have shown that the discrimination of bar orientation depends heavily upon the integrity of areas 17 and 18 (tier I). The present study indicates that several extrastriate areas in tiers II and III contribute to this discrimination task. Our data suggest that the anterior medial lateral suprasylvian, the posterior lateral lateral suprasylvian (tier II), and the anterior lateral lateral suprasylvian (tier III) areas are most likely to contribute to bar orientation discrimination.