Analysis of microsatellite instability using Promega panel in dermatofibrosarcoma protuberans.

Dermatofibrosarcoma protuberans (DFSP) is a rare neoplasm derived from fibroblasts. Although the frequency of microsatellite instability (MSI) in skin cancer is reported to be less than 5%, there is only one report of the status of MMR in DFSP. The only analytical report of microsatellite stability in which Promega panel is not used, showed that the frequency of MSI-high, MSI-low and microsatellite stable (MSS) cases was 13.9% (5/36), 16.7% (6/36) and 69.4% (25/36), respectively. Thus, the aim of this study was to evaluate the status of MMR in 36 patients with DFSP diagnosed at Kumamoto University. MSI analysis using the Promega panel showed that all cases were MSS, which indicated the absence of MSI in DFSP. This result indicates that the status of MMR may not be useful for the potential therapeutic application of pembrolizumab and the pathogenesis of DFSP may not involve MSI.

Dopamine D2 receptor supersensitivity in the hypothalamus of olfactory bulbectomized mice.

Olfactory bulbectomy (OBX) in rodents induces neurochemical and behavioral changes similar to those observed in individuals with depressive disorders. Our previous study suggested that OBX alters dopaminergic function in the striatum of mice; however, the effects on dopaminergic function in the hypothalamus is unknown. Therefore, in this study we examined dopaminergic system changes in the hypothalamus after OBX. Mice were administrated either the nonselective dopamine (DA) agonist apomorphine or the selective D2 agonist quinelorane, or pretreated with the selective D1 antagonist SCH23390 in combination with the selective D2 antagonist sulpiride or D3 antagonist SB277011A. Body temperature, which is regulated by the hypothalamic dopaminergic system, was monitored to evaluate changes in the dopaminergic system of the hypothalamus. DA D2 receptor (D2DR), tyrosine hydroxylase (TH), and phosphorylated (p)- DA- and cAMP-regulated phosphoprotein-32 (DARPP-32) levels in the hypothalamus were evaluated by western blotting. OBX mice exhibited significantly enhanced apomorphine-induced or quinelorane-induced hypothermia. The apomorphine-induced hypothermic response was reversed by the administration of sulpiride, but not SCH23390 or SB277011A. Moreover, TH and p-DARPP-32 levels were reduced and D2DR increased in the hypothalamus of OBX mice. These findings revealed that the OBX mice display enhanced DA receptor responsiveness associated with the hypothalamus, which may relate to some of the behavioral and neurochemical alterations reported in this animal model. Identification of changes in the hypothalamic dopaminergic system of OBX mice may provide useful information for the development of novel antidepressant treatments.

A study of EEG mu neurofeedback during action observation.

The mirror system is a brain network that gets activated during action performance and observation. Brain mu waves have been used as a mirror system activity index; however, mu rhythm is prone to contamination by occipital alpha wave activity, thus raising a concern regarding its reliability as an index of the mirror system activity. In this study, we investigated whether mu suppression can be used as an index of neurofeedback training, which influences mirror system activities. Participants observed videos of hand movement under three different conditions: central mu feedback (muFB), occipital alpha feedback (aFB), and simple observation without any feedback (OBS). Results showed that at the 4-5 min mark, mu wave was most significantly suppressed in the central site at muFB. We thus demonstrated the possibility of increasing mu wave suppression in feedback training using a specific stimulus such as motion observation.

Effect of Benifuuki Tea on Cytochrome P450-mediated Metabolic Activity in Rats.

BACKGROUND/AIM: Benifuuki tea has recently been used as an alternative therapy for pollinosis, and it may be consumed with pharmaceutical drugs. This study aimed to examine cytochrome P450 (CYP)-mediated food-drug interactions with Benifuuki tea in rats. MATERIALS AND METHODS: The inhibitory effects of Benifuuki tea and (-)-epigallocatechin-3-O-(3-O-methyl) gallate (EGCG3"Me) on CYP activities were evaluated in vitro. Midazolam pharmacokinetics was investigated after two treatments with Benifuuki tea. In an ex vivo study, CYP activities were determined after 1-week-treatment with the tea. RESULTS: Benifuuki tea and EGCG3"Me inhibited CYP2D and CYP3A activities in a concentration-dependent manner in vitro. However, MDZ metabolism did not change by Benifuuki treatment in vivo and ex vivo. In contrast, CYP2D activity was decreased ex vivo. CONCLUSION: Normal intake of Benifuuki tea is not likely to cause food-drug interactions by CYP3A inhibition or induction. In contrast, Benifuuki tea consumption may lead to food-drug interactions through the inhibition of CYP2D.

In vivo inhibition of CYP3A-mediated midazolam metabolism by anchusan in rats.

Cytochrome P450 (CYP)-mediated drug interactions caused by Kampo medicine have not been investigated sufficiently. The current study was conducted to reveal the effect of anchusan, a commonly used Kampo formula for gastrointestinal disease, on CYP3A-mediated drug metabolism in rats. The pharmacokinetics of midazolam (MDZ) was investigated after the single or one-week administration of anchusan (500 mg/kg) to evaluate its inhibitory and inducible effect on CYP3A, respectively. MDZ was administrated 16 h after the last anchsan treatment in the multiple dose study, while their intervals were 2 or 16 h in the single dose study. Unexpectedly, the multiple-pretreatment of anchusan increased the AUC of MDZ by 2.4-fold rather than decreasing it, and the CYP3A contents and activities were unchanged in hepatic and intestinal microsomes of these rats. In contrast, no significant inhibitory effects on MDZ metabolism were observed by the single anchusan pretreatment. In vitro study showed that the preincubation of anchusan and some of its component extracts with rat liver microsomes reduced CYP3A activity in a time- and NADPH-dependent manner. These results suggested that anchusan increased the serum MDZ concentration in rats, at least in part, by the time-dependent inhibition of CYP3A.

Effects of continuous ingestion of herbal teas on intestinal CYP3A in the rat.

Tenryocha, rooibos, and guava teas are widely consumed as herbal beverages, especially as a therapy against pollen allergy. To investigate the possible herbal tea-drug interaction the effect of continuous ingestion of these teas on cytochrome P450 (CYP) 3A were studied. Rats (n = 6) were allowed free access to either tea (experimental groups) or water (control) for two weeks. Midazolam (MDZ) (20 mg/kg) was orally administered and the serum concentration was determined. The area under the serum concentration-time curve (AUC(0-infinity)) and the maximum serum concentrations (C(max)) of MDZ were reduced by more than 60% after the treatment of tenryocha and rooibos tea (P<0.05). Intestinal MDZ 1'- and 4-hydroxylation activities mediated by CYP3A were increased in tenryocha and rooibos tea-treated group by 50% compared to the control group, although the results were not statistically significant. Furthermore, the Western blot analysis showed that CYP3A content was significantly increased in the intestine after the treatment of these teas (P<0.05). Hepatic MDZ hydroxylation and CYP3A content were slightly increased by these teas. The results suggested that two weeks ingestion of tenryocha and rooibos tea reduced serum concentration of MDZ by the induction of intestinal CYP3A. The possible interaction between tenryocha or rooibos tea and medicines mediated by CYP3A was suggested.

Surface plasmon resonance analysis for the screening of anti-prion compounds.

The interaction of anti-prion compounds and amyloid binding dyes with a carboxy-terminal domain of prion protein (PrP121-231) was examined using surface plasmon resonance (SPR) and compared with inhibition activities of abnormal PrP formation in scrapie-infected cells. Most examined compounds had affinities for PrP121-231: antimalarials had low affinities, whereas Congo red, phthalocyanine and thioflavin S had high affinities. The SPR binding response correlated with the inhibition activity of abnormal PrP formation. Several drugs were screened using SPR to verify the findings: propranolol was identified as a new anti-prion compound. This fact indicates that drug screenings by this assay are useful.