Histone deacetylase 6 inhibition enhances oncolytic viral replication in glioma

Oncolytic viral (OV) therapy, which uses genetically engineered tumor-targeting viruses, is being increasingly used in cancer clinical trials due to the direct cytolytic effects of this treatment that appear to provoke a robust immune response against the tumor. As OVs enter tumor cells, intrinsic host defenses have the potential to hinder viral replication and spread within the tumor mass. In this report, we show that histone deacetylase 6 (HDAC6) in tumor cells appears to alter the trafficking of post-entry OVs from the nucleus toward lysosomes. In glioma cell lines and glioma-stem–like cells, HDAC6 inhibition (HDAC6i) by either pharmacologic or genetic means substantially increased replication of oncolytic herpes simplex virus type 1 (oHSV). Moreover, HDAC6i increased shuttling of post-entry oHSV to the nucleus. In addition, electron microscopic analysis revealed that post-entry oHSVs are preferentially taken up into glioma cells through the endosomal pathway rather than via fusion at the cell surface. Together, these findings illustrate a mechanism of glioma cell defense against an incoming infection by oHSV and identify possible approaches to enhance oHSV replication and subsequent lysis of tumor cells.     

Developing a novel custom cutting guide for curved peri-acetabular osteotomy

Purpose

Curved peri-acetabular osteotomy (CPO) produces excellent clinical results, but the surgical procedure is technically demanding, and severe complications related to the osteotomy have been reported. To provide a safe, accurate surgical procedure, we have developed a novel method for setting the cutting line and direction. We have designed and made a custom cutting guide for individual patients. The purpose of the study was to evaluate the efficacy of this new method and cutting guide.

Methods

The cutting line was designed on a full-scale three-dimensional plaster model made from computed tomography (CT) data for each case. The surface of each plaster model was colour-coded according to the distance from the centre of the femoral head. A custom cutting guide was designed based on this cutting line on the workstation. A titanium custom cutting guide was fabricated using rapid prototyping technology. The cutting guide directed the cutting direction of the osteotome. We evaluated the outcomes for seven consecutive hips in seven patients who underwent CPO using the system between April and December 2011. All peri-operative complications were recorded. The accuracy of the cutting line was evaluated using CT data obtained two weeks after the operation.

Results

There were no major complications related to the osteotomy such as posterior column fracture or intra-articular osteotomy. The actual cutting line corresponded almost exactly to the planned cutting line in all cases.

Conclusions

The colour-coded plaster model and the custom cutting guide were effective for avoiding severe complications associated with a CPO.     

3D evaluation of composite resin restoration at practical training using swept-source optical coherence tomography (SS-OCT)

Internal adaptation of restorations to the cavity wall is one of the important topics in clinical dentistry. The purpose of this study was to investigate the possibility to utilize the non-invasive tomographic imaging system for teaching the importance of cavity adaptation at dental school pre-clinical training. Swept-source optical coherence tomography (SS-OCT) was used for detection of marginal and internal defects in the composite resin restorations as an educational device. Class 1 and Class 2 composite restorations to melamine resin molar tooth were assigned to the students and prepared during the skill test, and SS-OCT imaging was performed to evaluate students' works. SS-OCT could detect the internal gaps and voids within the restorations in tomography images synthesized based on the backscatter signal from within the restoration. It is suggested that the SS-OCT is promising diagnostic modality, as well as educational imaging device for the detection of internal gaps in adhesive restorations.     

Acupuncture stimulation to the sacral segment affects state of vigilance in rats

The effects of acupuncture stimulation to the sacral segment on electroencephalograms (EEGs) and activity of locus coeruleus (LC) neurons were examined in urethane-anesthetized rats. In 71 of 112 trials, when EEGs displayed small amplitude and high frequency, stimulation to the sacral segment-induced large amplitude and slow EEGs with a latency of <450s and duration ranged from 32s to >42 min. Stimulus-induced EEGs comprised significant increases in delta power and significant decreases in theta and beta powers. After intraperitoneal administration of bicuculline, stimulation to the sacral segment failed to induce changes in EEG pattern. Firing rate of noradrenergic LC neurons decreased significantly from 2.9+/-1.5 to 1.1+/-0.8 Hz (n=11, p<0.001). Decreased neuronal activity exhibited close relationships with increased EEG amplitude. These results suggest that acupuncture stimulation to the sacral segment changes the state of animals from light anesthesia to deep anesthesia, and that this change is mediated by GABAergic systems suppressing the activity of noradrenergic LC neurons.     

An acute dysfunction of the glutamate transport activity has been shown to generate free radicals and suppress the anti-oxidant ability in the hippocampus of rats

In this study, we attempted to elucidate whether or not an acute inhibition of glutamate transports activity with l-trans-pyrrolidine-2,4-dicarboxylic acid (l-trans PDC) would cause neuroexcitoxicity in the hippocampus. We used in vivo microdialysis and X-band electron spin resonance (ESR) spectroscopy to measure the changes in the redox state during the perfusion of l-trans PDC. ESR signals from rats using l-trans PDC were characteristically a six-line spectra, for which the hfc was a(N)=1.57mT and a(H)=0.25mT; these hfc's were obtained from the lipoxygenase/linoleic acid system that was used for the generation of lipid radicals. The antioxidant effect was measured using an ESR analysis to monitor sequential changes in the signal amplitude of nitroxide radical in the dialysate of both l-trans PDC and control animals. The pattern showed exponential decay with median half-life of the nitroxide radical took significantly longer in the l-trans PDC group. Acute changes in the glutamate transport resulted in the generation of a lipid radical and a depletion in the anti-oxidant effect in the hippocampus. Our data indicate that a dysfunction of a glutamate transport resulted in the collapse of the redox state, which thus eventually led to neuronal necrosis in the hippocampus. This study provides clear evidence for the mechanisms associated with neuronal disorder in relation to glutamate.     

Functional role of GABA transporters for kindling development in GLAST KO mice

Kindling-induced after discharge in electroencephalograms depends on the protein associated with glutamatergic and/or GABAergic neuronal transmission. In glutamate transporter knockout (GLAST KO) mice, the kindling phenomena in GLAST KO developed more slowly while the after discharge duration (ADD) was briefer than that of the control C57BL-6J mice. These findings indicate that either the excitatory function was suppressed or the inhibitory function was enhanced in GLAST KO kindling. To explain these phenomena, we used Western blotting to evaluate the alterations in the expression of hippocampal GABA transporter proteins, and the estimation of the effect on the process of epileptogenesis. Although no alterations were observed in the GAT-3 expression, the hippocampal GAT-1 expression was significantly suppressed in comparison to that of C57BL-6J mice. A decreased GAT-1 level in the hippocampus, which might be associated with the increased extracellular GABA level, may therefore inhibit both ADD and seizure propagation as shown by the amygdaloid kindling phenomenon observed in GLAST KO mice.     

Age-dependent changes in the hippocampal antioxidant ability of EL mice

Electron spin resonance (ESR) spectroscopy combined with in vivo microdialysis was used to analyze the antioxidant ability in the hippocampus of mice in an interictal state of EL mice utilizing decay ratio of an exogenously applied nitroxide radical (3-methoxycarbonyl-2,2,5,5-tetramethylpyrrolidine-1-oxyl (PCAM)). In EL mice with a history of frequent seizures, the half-life of the electron paramagnetism of PCAM in the hippocampus was prolonged. These results revealed decreased antioxidant ability, suggesting vulnerability against oxidative stress. Our data suggest that epileptogenesis in EL mice with chronic seizures is associated with functional failure due to the oxidized redox state and revealed that the decreased hippocampal antioxidant ability is related to the regional vulnerability to oxidative stress in the limbic system of EL mice during epileptogenesis.