Study on the relationship between the dispersal of wild boar (Sus scrofa) and the associated variability of Cesium-137 concentrations in its muscle Post-Fukushima Daiichi Nuclear Power Plant accident.

After the Fukushima Daiichi Nuclear Power Plant (FDNPP) accident in 2011, the wild boar (Sus scrofa) population within the Fukushima Evacuation Zone (FEZ) increased substantially in size and distribution. This growing population and their potential dispersal from the FEZ, where they are exposed to high levels of radionuclides, into the surrounding landscape underscores the need to better understand boar movement patterns in order to establish policies for managing shipping restrictions for boar meat and develop management strategies. In this study, we quantified the genetic population structure of boar in and around Fukushima prefecture using sequence data of the mitochondrial DNA control region and MIG-seq analysis using 348 boar samples to clarify boar dispersal patterns. Among boar samples, seven Asian haplotypes and one European haplotype were detected. The European haplotype originated from hybridization between domestic pigs and native boar in the evacuation zone after the accident and was detected in 15 samples across a broad geographic area. Our MIG-seq analysis revealed genetic structure of boar was significantly different between boar inhabiting the eastern (including FEZ. i.e., East clade) and western (i.e., West clade) regions in Fukushima prefecture. In addition, we investigated the relationships between boar dispersal and Cesium (Cs)-137 activity concentrations in boar muscle using MIG-seq genetic data in Nihonmatsu city, located in the central-northern region of Fukushima. High Cs-137 activity concentrations, exceeding 1000 Bq/kg, in boar muscle had a significantly high probability of belonging to the East clade within localized regions. Thus, our results provide evidence of the spatial scale of dispersal of individuals or offspring of boar from the FEZ. Results of this research also indicate that dispersal of individuals between areas with different Cs-137 contamination levels is one of the biggest factors contributing to variation in Cs-137 activity concentration in boar muscle within localized regions.

[An autopsied patient with palatal tremor and fatal bilateral vocal cord abduction paralysis associated with bilateral cerebellar dentate nucleus infarction].

A 74-year-old male patient developed multiple infarcts of the brainstem and cerebellum, followed 14 months later by palatal tremor and bilateral vocal cord abduction paralysis, resulting in death due to type 2 respiratory failure. Pathologic analysis revealed old infarcts extending from the bilateral cerebellar cortices to the dentate nucleus, being more extensive on the right side, accompanied by Wallerian degeneration involving the left red nucleus, right central tegmentum tract, and inferior cerebellar peduncle, followed by pseudohypertrophy of the bilateral inferior olivary nuclei. These lesions, involving the Guillain-Mollaret triangle, may have been responsible for the palatal tremor. On the other hand, there were no evident causative lesions for the vocal cord abduction, including any in the nucleus ambiguus or posterior cricoarytenoid muscles. In this case it is possible that the dysfunction responsible for the palatal tremor may have affected the pathway from the central tegmentum tract, which is part of the Guillain-Mollaret triangle, to the vagus nerve arising from the nucleus ambiguus, which plays a role in vocal cord abduction, thus affecting the vocal cords and resulting in abduction paralysis.

V180I genetic Creutzfeldt-Jakob disease: Severe degeneration of the inferior olivary nucleus in an autopsied patient with identification of the M2T prion strain.

Genetic Creutzfeldt-Jakob disease (gCJD) with a V180I mutation (V180I gCJD) is the most common type of gCJD in Japan, characterized by an older age at onset, slower progression, and moderate to severe cortical degeneration with spongiform changes and sparing of the brainstem and cerebellum. Degeneration of the inferior olivary nucleus (IO) is rarely observed in patients with CJD but is known to occur in fatal familial insomnia (FFI) and MM2-thalamic-type sporadic CJD (sCJD-MM2T) involving type 2 prion protein (M2T prion). Here we report on an 81-year-old Japanese woman who initially developed depressive symptoms followed by progressive cognitive impairment, myoclonus, and hallucinations and died after a clinical course of 23 months. Insomnia was not evident. Genetic analysis of the prion protein (PrP) identified a V180I mutation with methionine/valine heterozygosity at codon 129. Pathologic analysis demonstrated extensive spongiform degeneration, neuronal loss in the cortices, and weak synaptic-type PrP deposition. Except for IO degeneration, the clinicopathologic features and Western blotting PrP band pattern were compatible with those of previously reported V180I gCJD cases. Quantitative analysis revealed that the neuronal density of the IO, especially in the dorsal area, was considerably reduced to the same extent as that of a patient with sCJD-MM2T but preserved in other patients with V180I gCJD and sCJD-MM1 (this patient, 2.3 ± 0.53/mm2 ; a patient with sCJD-MM2T, 4.2 ± 2; a patient with V180I gCJD, 60.5 ± 9.3; and a patient with sCJD-MM1, 84.5 ± 17.9). Use of the protein misfolding cyclic amplification (PMCA) method confirmed the presence of the M2T prion strain, suggesting that the latter might be associated with IO degeneration in V180I gCJD. Autopsy studies are necessary to better understand the nature of CJD, since even if patients present with the common clinical picture, pathologic analysis might provide new insights, as was the case here.

Spinocerebellar ataxia type 17-digenic TBP/STUB1 disease: neuropathologic features of an autopsied patient.

Spinocerebellar ataxia (SCA) type 17-digenic TBP/STUB1 disease (SCA17-DI) has been recently segregated from SCA17, caused by digenic inheritance of two gene mutations - intermediate polyglutamine-encoding CAG/CAA repeat expansions (polyQ) in TBP (TBP41 - 49) and STUB1 heterozygosity - the former being associated with SCA17, and the latter with SCA48 and SCAR16 (autosomal recessive). In SCA17, most patients carry intermediate TBP41 - 49 alleles but show incomplete penetrance, and the missing heritability can be explained by a new entity whereby TBP41 - 49 requires the STUB1 variant to be symptomatic. The STUB1 gene encodes the chaperone-associated E3 ubiquitin ligase (CHIP) involved in ubiquitin-mediated proteasomal control of protein homeostasis. However, reports of the neuropathology are limited and role of STUB1 mutations in SCA17-DI remain unknown. Here we report the clinicopathologic features of identical twin siblings, one of whom was autopsied and was found to carry an intermediate allele (41 and 38 CAG/CAA repeats) in TBP and a heterozygous missense mutation in STUB1 (p.P243L). These patients developed autosomal recessive Huntington's disease-like symptoms. Brain MRI showed diffuse atrophy of the cerebellum and T2WI revealed hyperintense lesions in the basal ganglia and periventricular deep white matter. The brain histopathology of the patient shared features characteristic of SCA17, such as degeneration of the cerebellar cortex and caudate nucleus, and presence of 1C2-positive neurons. Here we show that mutant CHIP fails to generate the polyubiquitin chain due to disrupted folding of the entire U box domain, thereby affecting the E3 activity of CHIP. When encountering patients with cerebellar ataxia, especially those with Huntington's disease-like symptoms, genetic testing for STUB1 as well as TBP should be conducted for diagnosis of SCA17-DI, even in cases of sporadic or autosomal recessive inheritance.

Low cerebrospinal fluid-to-plasma ratios of orally administered lenalidomide mediated by its low cell membrane permeability in patients with hematologic malignancies.

Lenalidomide is a synthetic analog of thalidomide formed by the removal of one keto group (plus the addition of an amino group); it has anti-tumor activities beneficial for the treatment of hematologic malignancies. However, lenalidomide distribution to brain in animal models is reportedly low compared with that of thalidomide. The aim of this study was to evaluate plasma and cerebrospinal fluid concentrations of lenalidomide in three patients with malignant hematologic malignancies. Lenalidomide was detected in plasma from the three Japanese patients 1.5 h following oral administration of 20 mg lenalidomide using liquid chromatography/mass spectrometry, despite the in vitro gastrointestinal permeability of lenalidomide being low. Clinically observed cerebrospinal fluid-to-plasma ratios of lenalidomide were low (1.3-2.4%). Observed influx permeability values for lenalidomide in monkey blood-brain barrier model and human placental cell systems were one order of magnitude lower than those of thalidomide and another second-generation drug, pomalidomide along with a positive permeability control, caffeine. Because of the low cell-barrier permeability of lenalidomide demonstrated in in vitro assays, clinically relevant pharmacokinetic profiles of lenalidomide resulted in low penetrability from plasma into cerebrospinal fluid in patients with hematologic malignancies. Lenalidomide is conclusively suggested to expert its favorable immunomodulatory effects via systemic exposures in the patients.

Radiocesium-bearing microparticles cause a large variation in 137Cs activity concentration in the aquatic insect Stenopsyche marmorata (Tricoptera: Stenopsychidae) in the Ota River, Fukushima, Japan.

After the Tokyo Electric Power Company Fukushima Daiichi Nuclear Power Plant accident in Japan, freshwater ecosystems near the site remained contaminated by radiocesium (RCs). Clarifying RCs concentrations in aquatic insects is crucial because fishes consume these insects that transfer RCs into freshwater ecosystems. As aquatic insects are usually measured for radioactivity in bulk samples of several tens of insects, variation in RCs concentration among individuals is not captured. In this study, we investigated the variability in 137Cs activity concentration in individual aquatic insects in detritivorous caddisfly (Stenopsyche marmorata) and carnivorous dobsonfly (Protohermes grandis) larvae from the Ota River, Fukushima. Caddisfly larvae showed sporadically higher radioactivity in 4 of the 46 caddisfly larvae, whereas no such outliers were observed in 45 dobsonfly larvae. Autoradiography and scanning electron microscopy analyses confirmed that these caddisfly larvae samples contained radiocesium-bearing microparticles (CsMPs), which are insoluble Cs-bearing silicate glass particles. CsMPs were also found in potential food sources of caddisfly larvae, such as periphyton and drifting particulate organic matter, indicating that larvae may ingest CsMPs along with food particles of similar size. Although CsMP distribution and uptake by organisms in freshwater ecosystems is relatively unknown, our study demonstrates that CsMPs can be taken up by aquatic insects.

Monitoring of radioactive cesium in wild boars captured inside the difficult-to-return zone in Fukushima Prefecture over a 5-year period.

Following the Fukushima Daiichi Nuclear Power Plant accident in 2011, tissue samples from wild boar (Sus scrofa) outside the evacuation zone (difficult-to-return zone, DRZ) tended to show high activity concentrations of cesium-137 (137Cs). Understanding the 137Cs dynamics of wild boar populations inside the DRZ is necessary because they affect 137Cs dynamics and wild boar management in areas outside the DRZ. Since few detailed, long-term studies have been conducted inside the DRZ, we measured 137Cs activity concentrations in 221 wild boar muscle samples obtained from wild boar caught inside the DRZ and surrounding areas over a 5-year period. Our results showed that the 137Cs activity concentration in wild boar from inside the DRZ were higher than those in wild boar outside this zone. No significant difference was observed between muscle and soil 137Cs levels, but significant correlations were observed between muscle 137Cs activity concentrations and body length and body weight in the low-activity-concentration season, but not between all seasons and the high-activity-concentration seasons. It is considered that the size effects observed during the low-activity-concentration season may be due to factors related to metabolism and changes in food habit. This is the first long-term survey of 137Cs in wild boar inside the DRZ.

Hormone-sensitive lipase protects adipose triglyceride lipase-deficient mice from lethal lipotoxic cardiomyopathy.

Lipid droplets (LDs) are multifunctional organelles that regulate energy storage and cellular homeostasis. The first step of triacylglycerol hydrolysis in LDs is catalyzed by adipose triglyceride lipase (ATGL), deficiency of which results in lethal cardiac steatosis. Although hormone-sensitive lipase (HSL) functions as a diacylglycerol lipase in the heart, we hypothesized that activation of HSL might compensate for ATGL deficiency. To test this hypothesis, we crossed ATGL-KO (AKO) mice and cardiac-specific HSL-overexpressing mice (cHSL) to establish homozygous AKO mice and AKO mice with cardiac-specific HSL overexpression (AKO+cHSL). We found that cardiac triacylglycerol content was 160-fold higher in AKO relative to Wt mice, whereas that of AKO+cHSL mice was comparable to the latter. In addition, AKO cardiac tissues exhibited reduced mRNA expression of PPARα-regulated genes and upregulation of genes involved in inflammation, fibrosis, and cardiac stress. In contrast, AKO+cHSL cardiac tissues exhibited expression levels similar to those observed in Wt mice. AKO cardiac tissues also exhibited macrophage infiltration, apoptosis, interstitial fibrosis, impaired systolic function, and marked increases in ceramide and diacylglycerol contents, whereas no such pathological alterations were observed in AKO+cHSL tissues. Furthermore, electron microscopy revealed considerable LDs, damaged mitochondria, and disrupted intercalated discs in AKO cardiomyocytes, none of which were noted in AKO+cHSL cardiomyocytes. Importantly, the life span of AKO+cHSL mice was comparable to that of Wt mice. HSL overexpression normalizes lipotoxic cardiomyopathy in AKO mice and the findings highlight the applicability of cardiac HSL activation as a therapeutic strategy for ATGL deficiency-associated lipotoxic cardiomyopathies.

Genetic Population Structure of Wild Boars (Sus scrofa) in Fukushima Prefecture.

We aimed to reveal the dispersal and gene flow of the local wild boar (Sus scrofa) population and find their genetic boundary in Fukushima Prefecture. After the nuclear incident in 2011, the land was considered a difficult-to-return zone, and the increase in the number of wild boars was pronounced. To provide an effective management strategy for the wild boar population, we used multiplexed inter-simple sequence repeat genotyping by sequencing (MIG-seq) and clarified the genetic structure of wild boars. We obtained 328 single-nucleotide polymorphisms from 179 samples. STRUCTURE analysis showed that the most likely number of population cluster was K = 2. Molecular analysis of variance showed significant genetic differences between groups of wild boars inhabiting in the east and west across the Abukuma River. The migration rate from the eastern population to the western population is higher than in the reverse case based on BayesAss analysis. Our study indicates that both the Abukuma River and anthropogenic urbanization along the river may affect the migration of wild boars and the population in western was established mainly by the migration from other neighboring prefectures.

Candesartan prevents arteriopathy progression in cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy model.

Cerebral small vessel disease (CSVD) causes dementia and gait disturbance due to arteriopathy. Cerebral autosomal recessive arteriopathy with subcortical infarcts and leukoencephalopathy (CARASIL) is a hereditary form of CSVD caused by loss of high-temperature requirement A1 (HTRA1) serine protease activity. In CARASIL, arteriopathy causes intimal thickening, smooth muscle cell (SMC) degeneration, elastic lamina splitting, and vasodilation. The molecular mechanisms were proposed to involve the accumulation of matrisome proteins as substrates or abnormalities in transforming growth factor ß (TGF-ß) signaling. Here, we show that HTRA1-/- mice exhibited features of CARASIL-associated arteriopathy: intimal thickening, abnormal elastic lamina, and vasodilation. In addition, the mice exhibited reduced distensibility of the cerebral arteries and blood flow in the cerebral cortex. In the thickened intima, matrisome proteins, including the hub protein fibronectin (FN) and latent TGF-ß binding protein 4 (LTBP-4), which are substrates of HTRA1, accumulated. Candesartan treatment alleviated matrisome protein accumulation and normalized the vascular distensibility and cerebral blood flow. Furthermore, candesartan reduced the mRNA expression of Fn1, Ltbp-4, and Adamtsl2, which are involved in forming the extracellular matrix network. Our results indicate that these accumulated matrisome proteins may be potential therapeutic targets for arteriopathy in CARASIL.

Prediction of permeability across intestinal cell monolayers for 219 disparate chemicals using in vitro experimental coefficients in a pH gradient system and in silico analyses by trivariate linear regressions and machine learning.

For medicines, the apparent membrane permeability coefficients (Papp) across human colorectal carcinoma cell line (Caco-2) monolayers under a pH gradient generally correlate with the fraction absorbed after oral intake. Furthermore, the in vitro Papp values of 29 industrial chemicals were found to have an inverse association with their reported no-observed effect levels for hepatotoxicity in rats. In the current study, we expanded our influx permeability predictions for the 90 previously investigated chemicals to both influx and efflux permeability predictions for 207 diverse primary compounds, along with those for 23 secondary compounds. Trivariate linear regression analysis found that the observed influx and efflux logPapp values determined by in vitro experiments significantly correlated with molecular weights and the octanol-water distribution coefficients at apical and basal pH levels (pH 6.0 and 7.4, respectively) (apical to basal, r = 0.76, n = 198; and basal to apical, r = 0.77, n = 202); the distribution coefficients were estimated in silico. Further, prediction accuracy was enhanced by applying a light gradient boosting machine learning system (LightGBM) to estimate influx and efflux logPapp values that incorporated 17 and 19 in silico chemical descriptors (r = 0.83-0.84, p < 0.001). The determination in vitro and/or prediction in silico of permeability coefficients across intestinal cell monolayers of a diverse range of industrial chemicals/food components/medicines could contribute to the safety evaluations of oral intakes of general chemicals in humans. Such new alternative methods could also reduce the need for animal testing during toxicity assessment.

Long Spinal Cord Lesions Caused by Venous Congestive Myelopathy Associated with Intravascular Large B-cell Lymphoma.

Intravascular large B-cell lymphoma (IVLBCL) is a subtype of B-cell lymphoma, characterized by lymphoma cell proliferation within small blood vessels. We herein describe a rare case with long spinal cord lesions caused by venous congestive myelopathy associated with IVLBCL. An 81-year-old man presented with paraplegia of the lower limbs and sensory disturbances. Magnetic resonance imaging revealed intramedullary longitudinal T2-hyperintensity lesions in the thoracic cords. The patient died three months after disease onset, and a neuropathological analysis revealed predominantly atypical B-lymphocytes located sparsely in the veins of the spinal cord. IVLBCL should be considered in the differential diagnoses of long spinal cord lesions.

Effects of 137Cs contamination after the TEPCO Fukushima Dai-ichi Nuclear Power Station accident on food and habitat of wild boar in Fukushima Prefecture.

After the Tokyo Electric Power Company Fukushima Dai-ichi Nuclear Power Station accident, wild boar was found to have greater radiocesium (137Cs) activity concentrations in their bodies than other wild animals in Japan; however, the reason for this remains unknown. To understand the mechanism of 137Cs transfer from the environment to wild boar, and the factors that affect variation in 137Cs contamination in wild boar, we sampled muscle and stomach contents from wild boar captured in Fukushima Prefecture and analyzed the relationships among 137Cs concentrations in muscle tissue and in the stomach contents, 137Cs ground deposition at capture sites, and wild boar food habits. Significant positive relationships were observed among 137Cs activity concentrations in muscle and stomach contents, as well as 137Cs deposition density at capture sites. These results suggest that 137Cs is transferred from the environment to plant and animal materials consumed by wild boar, and then from these foods to the bodies of wild boar through digestion. However, no correlation was observed between 137Cs concentrations in stomach contents and the presence of any particular food item in stomachs of wild boar, including mushrooms. These findings suggest mushrooms and underground food items, which were found to affect 137Cs concentrations in wild animals in Europe, were not important contributors to high levels of 137Cs contamination in Japanese wild animals.

Relationship between radiocaesium in muscle and physicochemical fractions of radiocaesium in the stomach of wild boar.

After the accident at the TEPCO Fukushima Daiichi Nuclear Power Plant in 2011, it became important to study radiation dynamics, assess internal radiation exposure and specify factors affecting radionuclide variation in wildlife. Therefore, it is necessary to investigate which physicochemical fractions of radiocaesium (137Cs) are absorbed from ingested material in species with high activity concentrations of 137Cs, such as wild boar. This study analysed the physicochemical fractions of 137Cs in the stomach contents of wild boar to evaluate the transfer from ingested food to muscle. The 137Cs activity concentration in muscle showed a significantly positive relationship with the 137Cs activity concentration in the exchangeable fraction, and the sum of the 137Cs activity concentrations in the exchangeable and bound to organic matter fractions. Seasonal variations were also found in the 137Cs activity concentration in the exchangeable fraction, and the sum of the 137Cs activity concentrations in the exchangeable and bound to organic matter fractions. These findings suggest that the proportions of the physicochemical fractions of 137Cs in the exchangeable and bound to organic matter fractions in the stomach contents are important factors affecting the increases and seasonal dynamics of the activity concentrations of 137Cs in wild boar muscle.

Dramatic response of BRAF V600E-mutant epithelioid glioblastoma to combination therapy with BRAF and MEK inhibitor: establishment and xenograft of a cell line to predict clinical efficacy.

Epithelioid glioblastoma is a rare aggressive variant of glioblastoma (GBM) characterized by a dismal prognosis of about 6 months and frequent leptomeningeal dissemination. A recent study has revealed that 50% of epithelioid GBMs harbor three genetic alterations - BRAF V600E mutation, TERT promoter mutations, and homozygous deletions of CDKN2A/2B. Emerging evidence support the effectiveness of targeted therapies for brain tumors with BRAF V600E mutation. Here we describe a dramatic radiographical response to combined therapy with BRAF and MEK inhibitors in a patient with epithelioid GBM harboring BRAF V600E mutation, characterized by thick spinal dissemination. From relapsed tumor procured at autopsy, we established a cell line retaining the BRAF V600E mutation, TERT promoter mutation and CDKN2A/2B loss. Intracranial implantation of these cells into mice resulted in tumors closely resembling the original, characterized by epithelioid tumor cells and dissemination, and invasion into the perivascular spaces. We then confirmed the efficacy of treatment with BRAF and MEK inhibitor both in vitro and in vivo. Epithelioid GBM with BRAF V600E mutation can be considered a good treatment indication for precision medicine, and this patient-derived cell line should be useful for prediction of the tumor response and clarification of its biological characteristics.