Repeated spinal cord stimulation decreases the extracellular level of γ-aminobutyric acid in the periaqueductal gray matter of freely moving rats

Most of the previous experimental studies on the antinociceptive effects of electrical spinal cord stimulation (SCS) have focused on short-lasting effects mainly depending on spinal mechanisms. However, patients treated with SCS for chronic pain often report pain relief exceeding the period of stimulation for several hours. The long lasting effect of SCS might not only involve spinal, but also supraspinal mechanisms. A supraspinal region of major importance for the coordination of descending pain inhibition is the periaqueductal grey matter (PAG). The aim of the present microdialysis study, performed in awake freely moving rats, was to investigate if repeated SCS (two 30 min periods separated by a 90 min resting period) alters the extracellular neurotransmitter concentrations in the ventrolateral PAG. In a first series of experiments significantly decreased (−30%;P < 0.05;n = 7) γ-aminobutyric acid (GABA) levels were detected immediately after the second SCS session. Neither the concentration of serotonin nor that of substance P-like immunoreactivity (SP-LI) was affected by SCS. The decrease of GABA after two SCS sessions was confirmed in a second series of experiments (−30%;P < 0.05;n = 7). No spontaneous decline of GABA was observed in sham-stimulated animals (n = 6). The glutamate concentration was also determined in this latter series of experiments and a significant decrease (−23%;P < 0.05;n = 5) was observed after the second SCS session. As GABA-neurons in the PAG exert a tonic depressive effect on the activity in descending pain inhibitory pathways, a decreased extracellular GABA level in this region, as detected following repeated SCS, might indicate an increased pain inhibition.     

Anti-tumor effect of pEgr-interferon-γ-endostatin gene-radiotherapy in mice bearing Lewis lung carcinoma and its mechanism

Background Gene-radiotherapy, the combination of gene therapy and radiation therapy, is a new paradigm for cancer treatment. To enhance anti-tumor effect of gene-radiotherapy, in this study we construct a radiationinducible dual-gene co-expression vector pEgr-interferon(IFN)-γ/- endostatin and studied the anti-tumor effect of pEgr-IFN-γ/-endostatin gene-radiotherapy in mice bearing Lewis lung carcinoma and its mechanism.Methods Gene recombinant technique was used to construct dual-gene co-expression plasmid pEgr-IFN-γ endostatin, and single-gene expression plasmid pEgr-IFN-γ and pEgr-endostatin. The plasmids packed by liposome were injected locally into the tumors of the mice, and the tumors were irradiated with 5 Gy X-ray 36 hours later. The tumor growth rate at different time and mean survival period of the mice were observed.Cytotoxic activity of splenic cytotoxic T-lymphocyte ( CTL), natural killer (NK) cell and tumor necrosis factor (TNF)-α secretion activity of peritoneal macrophages of the mice in various groups were evaluated 15 days after irradiation. The intratumor micro-vessel density was evaluated by immunohistochemical staining 10 days after irradiation.Results The tumorgrowthrate of the mice in dual-gene-radiotherapy group was significantly lower than those in control group, 5 Gy group and single-gene-radiotherapy group at different time after gene-radiotherapy, and the mean survival period of which was longer. Cytotoxic activity of splenic CTL, NK and TNF-α secretion activity of peritoneal macrophages of the mice in dual-gene-radiotherapy group were significantly higher than those in control group, 5 Gy X-ray irradiation group and pEgr-endostatin gene-radiotherapy group 15 days after irradiation. The intratumor micro-vessel density of the mice in dual-gene-radiotherapy group was significantly lower than those in control group, 5 Gy X-ray irradiation group and pEgr-IFN-γ/gene-radiotherapy group.Conclusion The anti-tumor effect of dual-gene-radiotherapy was significantly better than that of single-gene-radiotherapy by combining the enhancement of anti-tumor immunologic function induced by IFN-γ/with the antiangiogenesis function of endostatin.     

Carotenoids, retinol and tocopherols in blood: comparability between serum and plasma (Li-heparin) values

OBJECTIVE: To assess the comparability of concentrations of retinol, alpha- and gamma-tocopherols and individual carotenoids in serum and (Li-hep) plasma over a wide range of concentrations. MATERIAL AND METHODS: One hundred sixty-six pairs of samples (serum and lithium-heparin plasma) were analyzed by a quality-controlled HPLC method. Means and 95% confidence intervals, differences, interchangeability and the degree of agreement (Bland-Altman plot) were calculated. RESULTS: Distribution of all analytes in the two matrices are comparable and interchangeable with minor quantitative adjustments. Within the range of concentrations assessed, the degree of agreement was high, although some differences were observed for minor components with greater analytical imprecision. CONCLUSIONS: The results indicate an acceptable degree of agreement using either of the two matrices for the analytes assessed except possibly for minor blood components. For retinol and alpha-tocopherol, the comparability and interchangeability of results below the cut-off points for inadequacy need further confirmation.     

Mutation analysis should be performed to rule out gammac deficiency in children with functional severe combined immune deficiency despite apparently normal immunologic tests

To study the correlation between genotype and phenotype in x-linked SCID, we have characterized the presentation of 2 unrelated patients. Both had infections suggestive of immunodeficiency, but their immune function and lymphoid tissues were normal. They were found to have an identical R222C mutation in the gammac gene.     

Immunotherapy with Agonistic Anti-CD137： Two Sides of a Coin

CD137 (4-1BB),a member of the TNF receptor superfamily,is an inducible T cell costimulatory receptorprimarily expressed on activated CD4~+ and CD8~+ T cells.Agonistic monoclonal antibodies (mAbs) against CD137greatly enhance T cell-mediated immune responses against many types of tumors and viruses.Surprisingly,theseagonists also showed therapeutic effects in several autoimmune diseases.These findings suggest that in differentdisease environments,CD137 engagement with agonist mAb in vivo can diametrically modulate immuneresponse outcomes.Therefore,CD137 agonists represent a promising immunotherapeutic approach to a widearray of disparate immune disorders.However,CD137's potency in modulating immune response necessitatescaution when targeting CD137 clinically.Cellular & Molecular Immunology.2004;1(1):31-36.     

Species differences in contaminants in fish on and adjacent to the Oak Ridge Reservation, Tennessee

Risks to humans and other organisms from consuming fish have become a national concern in the USA. In this paper, we examine the concentrations of 137Cs, arsenic, beryllium, cadmium, lead, mercury, and selenium in three species of fish from two river reaches adjacent to the US Department of Energy's Oak Ridge Reservation in Tennessee. We were interested in whether there were species and locational differences in radiocesium and metal concentrations and whether concentrations were sufficiently high to pose a potential health risk to humans or other receptors. Striped bass (Morone saxatilis) were significantly larger than white bass (M. chrysops), and crappie (Pomoxis spp.) were the smallest fish. Lead was significantly lower in striped bass, mercury was significantly higher in striped bass, and selenium was significantly higher in white bass compared to the other species. There were no other species differences in contaminants. White bass, the only species that was sufficiently abundant for a comparison, had significantly higher concentrations of cadmium, lead, and selenium in fillets from the Clinch River and significantly higher concentrations of mercury in fillets from Poplar Creek. The low concentrations of most contaminants in fish from the Clinch River do not appear to present a risk to humans or other consumers, although mercury concentrations in striped bass ranged as high as 0.79 ppm, well above the 0.5-ppm action level for human consumption of some US states.     

Acivicin-induced alterations in renal and hepatic glutathione concentrations and in gamma-glutamyltransferase activities

gamma-Glutamyltransferase (gamma-GT) catalyzes the hydrolysis of glutathione, glutathione S-conjugates, and gamma-substituted l-glutamate derivatives. Acivicin is an irreversible inhibitor of gamma-GT that has been used to study the role of gamma-GT in glutathione homeostasis and glutathione-dependent bioactivation reactions. The present studies were undertaken because of reported conflicting effects of acivicin on the nephrotoxicity of some haloalkenes that undergo glutathione-dependent bioactivation. The objective of this study was to test the hypothesis that acivicin may alter renal glutathione concentrations; acivicin-induced changes in renal glutathione concentrations may alter the susceptibility of the kidney to the nephrotoxic effects of haloalkenes. Hence, diurnal and acivicin-induced changes in renal and hepatic glutathione concentrations along with renal and hepatic gamma-GT activities were investigated. The previously observed diurnal variations in hepatic glutathione concentrations in fed rats were confirmed, but no diurnal variations were observed in renal glutathione concentrations or in renal or hepatic gamma-GT activities. Renal and hepatic glutathione concentrations and gamma-GT activities were measured in tissue homogenates from rats given 0, 0.1, or 0.2 mmol acivicin/kg (i.p.) and killed 0, 2, 4, 8, 12, or 24 hr later. Renal glutathione concentrations were increased above control values in acivicin-treated rats, whereas acivicin had no effect on hepatic glutathione concentrations. Renal gamma-GT activities decreased within 2 hr after giving acivicin and remained decreased for 24 hr. Acivicin had no effect on hepatic gamma-GT activities, except at 24 hr after treatment when values in acivicin-treated rats were elevated compared with controls. Although the present studies do not afford an explanation of the mechanism whereby acivicin increases the nephrotoxicity of some haloalkenes, they do indicate that acivicin is not a reliable probe to investigate the role of gamma-GT in haloalkene-induced nephrotoxicity.     

gamma-Diketone neuropathy: axon atrophy and the role of cytoskeletal protein adduction

Multifocal giant neurofilamentous axonal swellings and secondary distal degeneration have been historically considered the hallmark features of gamma-diketone neuropathy. Accordingly, research conducted over the past 25 years has been directed toward discerning mechanisms of axonal swelling. However, this neuropathological convention has been challenged by recent observations that swollen axons were an exclusive product of long-term 2.5-hexanedione (HD) intoxication at lower daily dose-rates (e.g., 175 mg/kg/day); that is, higher HD dose-rates (e.g., 400 mg/kg/day) produced neurological deficits in the absence of axonal swellings. The observation that neurological toxicity can be expressed without axonal swelling suggests that this lesion is not an important pathophysiological event. Instead, several research groups have now shown that axon atrophy is prevalent in nervous tissues of laboratory animals intoxicated over a wide range of HD dose-rates. The well-documented nerve conduction defects associated with axon atrophy, in conjunction with the temporal correspondence between this lesion and the onset of neurological deficits, strongly suggest that atrophy has pathophysiological significance. In this commentary, we present evidence that supports a pathognomonic role for axon atrophy in gamma-diketone neuropathy and suggests that the functional consequences of this lesion mediate the corresponding neurological toxicity. Previous research has demonstrated that HD interacts with proteins via formation of pyrrole adducts. We therefore discuss the possibility that this chemical process is essential to the mechanism of atrophy. Evidence presented in this review suggests that "distal axonopathy" is an inaccurate classification and future nosological schemes should be based on the apparent primacy of axon atrophy.     

NK and CD8+ T cell-mediated eradication of poorly immunogenic B16-F10 melanoma by the combined action of IL-12 gene therapy and 4-1BB costimulation

In previous reports, systemic administration of a stimulatory monoclonal antibody directed against the 4-1BB receptor had no effect on survival or tumor burden in mice inoculated with the poorly immunogenic B16-F10 melanoma. We combined IL-12 gene transfer with 4-1BB costimulation to explore a previously noted cooperative anti-tumor effect against this model tumor. We hypothesize that the innate immune response mediated by IL-12-activated natural killer (NK) cells initiates the activation of the immune system, leading to the priming of T cells, whereas 4-1BB costimulation enhances the function of primed tumor-specific T cells. The effect of the combination therapy on the growth of subcutaneous (s.c.) tumors and pulmonary metastasis was examined. The combination therapy significantly retarded the growth of subcutaneously-inoculated tumors, and 50% of tumor-bearing mice survived with complete tumor regression. In contrast, neither IL-12 gene transfer nor anti-4-1BB antibody administration alone was as effective. Enhanced CTL activity against both B16-F10 tumor cells and TRP-2-pulsed EL4 syngeneic tumor cells was observed in tumor-bearing animals treated with the combination therapy 2 weeks after treatment and, in long-term survivors from this combination therapy, at >120 days. In a pulmonary metastatic model, only the combination therapy generated significant protection against metastasis. In vivo depletion of NK or CD8(+) but not CD4(+) subsets eliminated the protective immunity. Furthermore, NK cell depletion significantly reduced both tumor-specific CTL activity and the number of tumor-specific IFN-gamma-producing cells, suggesting that this synergistic effect requires the participation of both NK and CD8(+) T cells.     

Sulfur in human crystallins

Molecular models of human gamma-crystallins and the 'alpha-crystallin domain' of human alphaA-crystallin have been built based on available related X-ray crystal structures. The accessibilities of the component cysteine, methionine and tryptophan side chains in the crystallin models have been calculated. The reactivities of these cysteines, which are oxidised in cataract, are assessed based on their known modifications and within the context of their location within the 3D models.