Increased Ca2+ channel currents in cerebellar Purkinje cells of the ataxic groggy rat

The ataxic groggy rat (strain name; GRY) is an autosomal recessive neurological mutant found in a closed colony of Slc:Wistar rats. Recent genetic analysis has identified the missense (M251K) mutation in the alpha(1) subunit of the Ca(V)2.1 (P/Q-type) voltage-dependent Ca(2+) channel gene (Cacna1a) of GRY rat. In this study, we found that high-voltage-activated (HVA) Ca(2+) channel currents in acutely dissociated Purkinje cells of GRY rats showed increased (not decreased) current density and depolarizing shift of the activation and inactivation curves compared with those of normal Wistar rats. In contrast low-voltage-activated (LVA) Ca(2+) channel currents of GRY rats showed no significant changes. These results suggest that functional alteration of Ca(2+) channel currents in cerebellar Purkinje cells of GRY rats is attributed to the change of HVA Ca(2+) channel currents, and that increased HVA Ca(2+) channel function underlies the cerebellar dysfunction and ataxic phenotype of GRY rats.     

Molecular cloning and functional characterization of porcine MyD88 essential for TLR signaling

We isolated cDNA encoding porcine MyD88 （poMyD88） from Peyer＇s patches （Pps） of GALT. The complete open reading frame （ORF） of poMyD88 contains 879 bp encoding a deduced 293 aa residues. The amino acid sequence of poMyD88 was characterized by N-terminal death, intermediate and C-terminal Toll/IL-1 receptor （TIR） domains. The putative poMyD88 protein shares a higher level of homology with its human （87.2% amino acid identity） than with its mouse （77.4% amino acid identity） counterpart. Overexpression of poMyD88 participated in the further enhanced activation of NF-w.B in human embryonic kidney （HEK） 293 cells expressing porcine TLR2 and porcine TLR4/MD-2, but not porcine RP105/MD-1 after stimulation with the corresponding ligands. The expression levels of MyD88 were highest in the spleen and mesenteric lymph nodes （MLNs）, and lower in digestive tissues of newborn swine. In adult swine, the expression levels in the digestive tissues were lower than those in MLNs and the spleen. These results suggest that an MyD88-dependent signaling pathway is present in newborn as well as in adult swine and that it is involved in the innate immune system of these animals.     

Evaluation of the Modified ELISPOT Assay for Gamma Interferon Production in Cancer Patients Receiving Antitumor Vaccines

Frequencies of vaccine-responsive T-lymphocyte precursors in peripheral blood mononuclear cells (PBMC) prior to and after administration of peptide-based vaccines in patients with cancer can be measured by limiting-dilution assays (LDA) or by ELISPOT assays. We have used a modified version of the ELISPOT assay to monitor changes in the frequency of gamma interferon (IFN-γ)-producing T cells in a population of lymphocytes responding to a relevant peptide or a nonspecific stimulator, such as phorbol myristate acetate-ionomycin. Prior to its use for monitoring of patient samples, the assay was validated and found to be comparable to the LDA performed in parallel, using tumor-reactive cytolytic T-lymphocyte (CTL) lines. The sensitivity of the ELISPOT assay was found to be 1/100,000 cells, with an interassay coefficient of variation of 15%, indicating that it could be reliably used for monitoring of changes in the frequency of IFN-γ-secreting responder cells in noncultured or cultured lymphocyte populations. To establish that the assay is able to detect the T-cell precursor cells responsive to the vaccine, we used CD8⁺ T-cell populations positively selected from PBMC of HLA-A2⁺ patients with metastatic melanoma, who were treated with dendritic cell-based vaccines containing gp100, MELAN-A/MART-1, tyrosinase, and influenza virus matrix peptides. The frequency of peptide-specific responder T cells ranged from 0 to 1/2,600 before vaccination and increased by at least 1 log unit after vaccination in two patients, one of whom had a clinical response to the vaccine. However, no increases in the frequency of peptide-responsive T cells were observed in noncultured PBMC or PBMC cultured in the presence of the relevant peptides after the melanoma patients enrolled in another trial were treated with the intramuscular peptide vaccine plus MF59 adjuvant. Thus, while the ELISPOT assay was found to be readily applicable to assessments of frequencies of CTL precursors of established CTL lines and ex vivo-amplified PBMC, its usefulness for monitoring of fresh PBMC in patients with cancer was limited. In many of these patients antitumor effector T cells are present at frequencies of lower than 1/100,000 in the peripheral circulation. Serial monitoring of such patients may require prior ex vivo amplification of specific precursor cells.     

Differences in short-term clinical and radiological outcomes depending on timing of balloon kyphoplasty for painful osteoporotic vertebral fracture

Background

Balloon kyphoplasty or vertebroplasty is widely performed as a surgical intervention for osteoporotic vertebral fracture (OVF) and the effects have been investigated in many previous studies. However, the influence of the timing of the procedure on patient outcomes has not been studied formally. The purpose of this study was to investigate differences in the surgical outcomes of OVFs according to the timing of balloon kyphoplasty.

Thalamic hypometabolism on 18FDG-positron emission tomography in medial temporal lobe epilepsy

OBJECTIVES: Degree of hypometabolism in the thalamus on (18)Fluorodeoxyglucose-positron emission tomography (FDG-PET) was compared with those of medial and lateral temporal lobes in patients with medial temporal lobe epilepsy (mTLE), and its relationship with post-operative seizure outcomes was investigated. METHODS: Twenty-six patients with mTLE who underwent anterior temporal lobectomy were included. Post-operatively, 13 patients became completely seizure-free and 13 showed residual seizure, regardless of frequency (five patients became almost seizure-free, six had rare seizures and two showed significant improvements). Degrees of hypometabolism in bilateral thalamus, ipsilateral medial and lateral temporal lobes were evaluated visually and semi-quantitatively by determining the asymmetry index (AI), a value indicating 100 x (ipsilateral - contralateral)/[1/2 x (ipsilateral + contralateral)] and the region-to-cerebral hemisphere ratio (R/C ratio) being the ratio between averaged counts in each area and those in the cerebral hemisphere of the same side. RESULTS: Hypometabolism in the medial temporal lobe was visually observed in all patients. Hypometabolism in the lateral temporal lobe was observed in 20 patients and was semi-quantitatively more prominent than that of the medial temporal lobe. Pathologically, hippocampal sclerosis and prominent astrogliosis of the lateral temporal lobe were present in all cases. However, while thalamic hypometabolism was visually observed in nine patients (in the ipsilateral side of four cases, contralateral side of three and on both sides of two), no significant thalamic hypometabolism was semi-quantitatively observed. No significant differences in metabolic rate in any area except for the lateral temporal lobe between seizure-free patients and residual seizure patients were seen semi-quantitatively. DISCUSSION: Data indicated that metabolism in the lateral temporal lobe of patients with mTLE significantly decreased and revealed pathologic glial changes. Thalamic hypometabolism was quite mild and did not correlate with post-operative seizure outcome.     

Synchronous monitoring of external/internal respiratory motion: validity of respiration-gated radiotherapy for liver tumors

Purpose  

Four-dimensional (4D) radiotherapy, in particular respiration gating for the treatment of lung tumors, is gaining popularity. Its utility for other sites, however, has not been investigated fully. The purpose of this study was to see whether 4D therapy is feasible for liver tumors.     

p38 mitogen-activated protein kinase inhibitor reduces neurocan production in cultured spinal cord astrocytes

Chondroitin sulfate proteoglycans are formed in scar tissue after a spinal cord injury and inhibit axon regrowth. The production of neurocan, one of these chondroitin sulfate proteoglycans, in cultured spinal cord astrocytes increased after the addition of epidermal growth factor (EGF) in a dose-dependent manner (2-200 ng/ml). In astrocytes stimulated by 20 ng/ml of EGF, neurocan production was inhibited after the addition of the p38 mitogen-activated protein kinase (MAPK) inhibitor (SB203580: 3-10 μM) in a dose-dependent manner. These results suggest that the activation of p38 MAPK is one of the mechanisms of neurocan production in EGF-stimulated astrocytes. The p38 MAPK inhibitor may reduce neurocan production and accelerate axonal regrowth after a spinal cord injury.