[Visualization of Effectiveness of the Medical Safety Measures -Suggestion of the Evaluation Technique for Rare Medical Incident].

For events with a low occurrence rate, such as medical incidents, we were able to determine the evaluation before and after taking medical safety measures by statistical methods (testing for differences in population rate). The point of this method is that we evaluated the occurrence rate of incidents to the total number of examinations (number of incident occurrence real number plus number of examinations carried out without any problems). Our results suggest that this technique becomes the evaluation technique as the effective method of medical safety measures. The present studies demonstrated that the evaluation technique by the testing for differences in population rate become the indicator to judge the effectiveness of the medical safety measures in the following cases. (1) When we evaluate the decrease in incident for the long term before and after safety measures. (2) When we evaluate the effectiveness of measures in the middle evaluation after safety measures.

[Examination of Ability to Support the Interpreting of CT Images of Emergency Transport Patients by the Radiological Technologist─About the Factors That Influences Image Interpretation Assistance─].

The Japanese Ministry of Health, Labor and Welfare announced a revision of the law about the expansion of duties by the radiological technologist in team medical care in April, 2010. In that respect, the importance of reading images with support from the radiological technologist became higher. We compared the interpretation results of the radiologist with the image analysis by the radiological technologist of the CT images of emergency patients. And we checked for symptoms and diseases which were frequently overlooked by the technologists. Inexperienced radiological technologists overlooked considerably more than experienced radiological technologists. Our results showed that abdominal or chest image analysis differed more often than head scan analysis. The reasons given for the differences include a lack of clear indication for abdominal tumors, and we overlooked a lot of diseases such as pneumonia and enteritis. We also had several cases of abdominal and chest images over-reading by radiological technologists. To improve these, radiological technologists should deepen their knowledge of normal anatomy and work to improve recalling diseases that are inferred from the patient's symptoms. This will greatly improve the image interpretation support by the radiological technologists.

GSK3B-mediated phosphorylation of MCL1 regulates axonal autophagy to promote Wallerian degeneration.

Macroautophagy is a catabolic process, in which portions of cytoplasm or organelles are delivered to lysosomes for degradation. Emerging evidence has indicated a pathological connection between axonal degeneration and autophagy. However, the physiological function and induction mechanism of autophagy in axons remain elusive. We herein show that, through activation of BECLIN1, glycogen synthase kinase 3B (GSK3B)-mediated phosphorylation of BCL2 family member MCL1 induces axonal autophagy and axonal degeneration. Phosphorylated MCL1 is ubiquitinated by the FBXW7 ubiquitin ligase and degraded by the proteasome, thereby releasing BECLIN1 to induce axonal autophagy. Axonal autophagy contributes to local adenosine triphosphate production in degenerating axons and the exposure of phosphatidylserine-an "eat-me" signal for phagocytes-on transected axons and is required for normal recruitment of phagocytes to axonal debris in vivo. These results suggest that GSK3B-MCL1 signaling to regulate autophagy might be important for the successful completion of Wallerian degeneration.

Glutamate signals through mGluR2 to control Schwann cell differentiation and proliferation.

Rapid saltatory nerve conduction is facilitated by myelin structure, which is produced by Schwann cells (SC) in the peripheral nervous system (PNS). Proper development and degeneration/regeneration after injury requires regulated phenotypic changes of SC. We have previously shown that glutamate can induce SC proliferation in culture. Here we show that glutamate signals through metabotropic glutamate receptor 2 (mGluR2) to induce Erk phosphorylation in SC. mGluR2-elicited Erk phosphorylation requires ErbB2/3 receptor tyrosine kinase phosphorylation to limit the signaling cascade that promotes phosphorylation of Erk, but not Akt. We found that Gßγ and Src are involved in subcellular signaling downstream of mGluR2. We also found that glutamate can transform myelinating SC to proliferating SC, while inhibition of mGluR2 signaling can inhibit demyelination of injured nerves in vivo. These data suggest pathophysiological significance of mGluR2 signaling in PNS and its possible therapeutic importance to combat demyelinating disorders including Charcot-Marie-Tooth disease.

Hepatic rRNA transcription regulates high-fat-diet-induced obesity.

Ribosome biosynthesis is a major intracellular energy-consuming process. We previously identified a nucleolar factor, nucleomethylin (NML), which regulates intracellular energy consumption by limiting rRNA transcription. Here, we show that, in livers of obese mice, the recruitment of NML to rRNA gene loci is increased to repress rRNA transcription. To clarify the relationship between obesity and rRNA transcription, we generated NML-null (NML-KO) mice. NML-KO mice show elevated rRNA level, reduced ATP concentration, and reduced lipid accumulation in the liver. Furthermore, in high-fat-diet (HFD)-fed NML-KO mice, hepatic rRNA levels are not decreased. Both weight gain and fat accumulation in HFD-fed NML-KO mice are significantly lower than those in HFD-fed wild-type mice. These findings indicate that rRNA transcriptional activation promotes hepatic energy consumption, which alters hepatic lipid metabolism. Namely, hepatic rRNA transcriptional repression by HFD feeding is essential for energy storage.

Na+/H+ exchangers induce autophagy in neurons and inhibit polyglutamine-induced aggregate formation.

In polyglutamine diseases, an abnormally elongated polyglutamine results in protein misfolding and accumulation of intracellular aggregates. Autophagy is a major cellular degradative pathway responsible for eliminating unnecessary proteins, including polyglutamine aggregates. Basal autophagy constitutively occurs at low levels in cells for the performance of homeostatic function, but the regulatory mechanism for basal autophagy remains elusive. Here we show that the Na(+)/H(+) exchanger (NHE) family of ion transporters affect autophagy in a neuron-like cell line (Neuro-2a cells). We showed that expression of NHE1 and NHE5 is correlated to polyglutamine accumulation levels in a cellular model of Huntington's disease, a fatal neurodegenerative disorder characterized by accumulation of polyglutamine-containing aggregate formation in the brain. Furthermore, we showed that loss of NHE5 results in increased polyglutamine accumulation in an animal model of Huntington's disease. Our data suggest that cellular pH regulation by NHE1 and NHE5 plays a role in regulating basal autophagy and thereby promotes autophagy-mediated degradation of proteins including polyglutamine aggregates.

Wallerian degeneration slow mouse neurons are protected against cell death caused by mechanisms involving mitochondrial electron transport dysfunction.

Ischemia elicits a variety of stress responses in neuronal cells, which result in cell death. wld(S) Mice bear a mutation that significantly delays Wallerian degeneration. This mutation also protects all neuronal cells against other types of stresses resulting in cell death, including ischemia. To clarify the types of stresses that neuronal cell bodies derived from wld(S) mice are protected from, we exposed primary cultured neurons derived from wld(S) mice to various components of hypoxic stress. We found that wld(S) mouse neurons are protected against cellular injury induced by reoxygenation following hypoxic stress. Furthermore, we found that wld(S) mouse neurons are protected against functional impairment of the mitochondrial electron transport chain. These data suggest that Wld(S) protein expression may provide protection against neuronal cell death caused by mechanisms involving mitochondrial electron transport dysfunction.

Biological in vitro and in vivo studies of a series of new asymmetrical cationic [99mTc(N)(DTC-Ln)(PNP)]+ complex (DTC-Ln = alicyclic dithiocarbamate and PNP = diphosphinoamine).

(99m)Tc(N)-DBODC5 is a cationic mixed compound under clinical investigation as potential myocardial imaging agent. In spite of this, analogously to the other cationic (99m)Tc-agents, presents a relatively low first-pass extraction. Thus, modification of (99m)Tc(N)-DBODC(5) direct to increase its first-pass extraction keeping unaltered the favorable imaging properties would be desirable. This work describes the synthesis and biological evaluation of a series of novel cationic (99m)Tc-nitrido complexes, of general formula [(99m)TcN(DTC-Ln)(PNP)](+) (DTC-Ln= alicyclic dithiocarbamates; PNP = diphosphinoamine), as potential radiotracers for myocardial perfusion imaging. The synthesis of cationic (99m)Tc-(N)-complexes were accomplished in two steps. Biodistribution studies were performed in rats and compared with the distribution profiles of (99m)Tc(N)-DBODC5 and (99m)Tc-Sestamibi. The metabolisms of the most promising compounds were evaluated by HPLC methods. Biological studies revealed that most of the complexes have a high initial and persistent heart uptake with rapid clearance from nontarget tissues. Among tested compounds, 2 and 12 showed improved heart uptake with respect to the gold standard (99m)Tc-complexes with favorable heart-to-liver and slightly lower heart-to-lung ratios. Chromatographic profiles of (99m)Tc(N)-radioactivity extracted from tissues and fluids were coincident with the native compound evidencing remarkable in vivo stability of these agents. This study shows that the incorporation of alicyclic dithiocarbamate in the [(99m)Tc(N)(PNP)](+) building block yields to a significant increase of the heart uptake at early injection point suggesting that the first-pass extraction fraction of these novel complexes may be increased with respect to the other cationic (99m)Tc-agents keeping almost unaltered the favorable target/nontarget ratios.

Subcellular distribution and metabolism studies of the potential myocardial imaging agent [99mTc(N)(DBODC)(PNP5)]+.

UNLABELLED: 99mTc(N)-DBODC5 is the lead compound of a new series of monocationic 99mTc(N)-based potential myocardial imaging agents that exhibit original biodistribution properties. This study was addressed to elucidate the mechanisms of distribution, retention, and elimination of this promising 99mTc(N)-agent. METHODS: The sex-related in vitro and in vivo stability and the subcellular distribution of 99mTc(N)-DBODC5 were investigated. Studies were performed by considering binding to the serum proteins; stability in rat serum, human serum, and rat liver homogenates; and the chemical integrity of the complex after extraction from rat tissues such as heart, liver, and kidney, as well as from intestinal fluids and urine. The effect of cyclosporin A on the in vivo pharmacokinetic properties of 99mTc(N)-DBODC5 was also evaluated. Subcellular distribution of 99mTc(N)-DBODC5 in ex vivo rat heart was determined by standard differential centrifugation techniques. RESULTS: No significant in vitro serum protein binding and no notable biotransformation of the native compound into different species by the in vitro action of the serum and liver enzymes was evidenced. In vivo experiments showed that sex affects the pharmacokinetic profile of the 99mTc(N)-complexes including metabolism and excretion. Chromatographic profiles of 99mTc(N)-radioactivity extracted from tissues and fluids of female rats were always coincident with the control. Conversely, a small percentage of metabolized species was detected by high-performance liquid chromatography in liver extracts of male rats. Furthermore, administration of cyclosporin A caused a significant reduction of lung, liver, and kidney washout along with a considerable variation in activity distribution in the intestinal tract in both male and female rats, thus indicating a possible implication of Pgp transporters in determining the biologic behavior of 99mTc(N)-DBODC5. However, this phenomenon was more pronounced in females. Subcellular distribution studies showed that 86.3% +/- 7.4% of 99mTc(N)-DBODC5 was localized into mitochondrial fraction as a result of the interaction with the negative membrane potential. CONCLUSION: Evidence showing that the new 99mTc(N)-myocardial tracers behave as multidrug resistance-associated protein P-glycoprotein substrates, combined with their selective mitochondrial accumulation, strongly supports the possibility that diagnostic application of 99mTc(N)-DBODC5 can be extended to tumor imaging and noninvasive multidrug resistance studies.