Renal macrophages induce hypertension and kidney fibrosis in Angiotensin II salt mice model.

The immune system is involved in hypertension development with different immune cells reported to have either pro or anti-hypertensive effects. In hypertension, immune cells have been thought to infiltrate blood pressure-regulating organs, resulting in either elevation or reduction of blood pressure. There is controversy over whether macrophages play a detrimental or beneficial role in the development of hypertension, and the few existing studies have yielded conflicting results. This study aimed to determine the effects of angiotensin II (Ang II) salt-induced hypertension on renal immune cells and to determine whether renal macrophages are involved in the induction of hypertension. Hypertension was induced by administration of Ang II and saline for two weeks. The effects of hypertension on kidney immune cells were assessed using flow cytometry. Macrophage infiltration in the kidney was assessed by immunohistochemistry and kidney fibrosis was assessed using trichrome stain and kidney real time-qPCR. Liposome encapsulated clodronate was used to deplete macrophages in C57BL/6J mice and investigate the direct role of macrophages in hypertension induction. Ang II saline mice group developed hypertension, had increased renal macrophages, and had increased expression of Acta2 and Col1a1 and kidney fibrotic areas. Macrophage depletion blunted hypertension development and reduced the expression of Acta2 and Col1a1 in the kidney and kidney fibrotic areas in Ang II saline group. The results of this study demonstrate that macrophages infiltrate the kidneys and increase kidney fibrosis in Ang II salt-induced hypertension, and depletion of macrophages suppresses the development of hypertension and decreases kidney fibrosis. This indicates that macrophages play a direct role in hypertension development. Hence macrophages have a potential to be considered as therapeutic target in hypertension management.

Thermal and Optical Properties of Porous Nanomesh Structures for Sensitive Terahertz Bolometric Detection.

Terahertz (THz) electromagnetic waves are attractive for use in nondestructive and biocompatible sensing applications. Thermal sensors are widely used for THz detection owing to the small photon energies of THz radiation, where this requires materials with low thermal conductivity and a small heat capacity to ensure the sensitive and fast operation of the sensors. In this study, we investigated the thermal and optical properties of porous nanomesh structures for sensitive THz bolometric detection. Nanometer (nm)-scale hole array structures were formed on gallium arsenide (GaAs) microelectromechanical system (MEMS) beams to improve their thermal properties. The thermal conductance of the porous MEMS beams was obtained by measuring their thermal bandwidths; it was found to decrease by as much as ~90% when the porosity (P) of the porous nanostructure was increased to ~0.69. We also measured the THz absorptance of the porous hole array structure. The results show that although the porous nanostructure has a much smaller area than the bulk material, it maintained a high coefficient of THz absorptance because the featured size was much smaller than the THz wavelength. The measured absorptance agreed well with that calculated by using the Drude model. These results demonstrate that the porous nanomesh structure is promising for developing highly sensitive THz thermal sensors.

Polydiacetylene Liposomal Aequorin Bioluminescent Device for Detection of Hydrophobic Compounds.

In this study, a polydiacetylene liposomal aequorin bioluminescent device (PLABD) that functioned through control of the membrane transport of Ca(2+) ions was developed for detecting hydrophobic compounds. In the PLABD, aequorin was encapsulated in an internal water phase and a calcium ionophore (CI) was contained in a hydrophobic region. Membrane transport of Ca(2+) ions across the CI was suppressed by polymerization between diacetylene molecules. On addition of an analyte, the membrane transport of Ca(2+) ions across the CI increased, and Ca(2+) ions from the external water phase could diffuse into the internal water phase via the CI, which resulted in bioluminescence of the aequorin. Lidocaine, procaine, and procainamide were used as model compounds to test the validity of the detection mechanism of the PLABD. When each analyte was added to a suspension of the PLABD, bioluminescence from the aequorin in the PLABD was observed, and the level of this bioluminescence increased with increasing analyte concentration. There was a linear relationship between the logarithm of the analyte concentration and the bioluminescence for all analytes as follows: R = 0.89 from 10 nmol L(-1) to 10 mmol L(-1) for lidocaine, R = 0.66 from 10 nmol L(-1) to 100 µmol L(-1) for procaine, and R = 0.74 from 100 nmol L(-1) to 100 µmol L(-1) for procainamide. Compared to the traditional colorimetric method using polydiacetylene liposome, the PLABD was superior for both the sensitivity and dynamic range. Thus, PLABD is a valid, simple, and sensitive signal generator for detection of hydrophobic compounds that interact with PLABD membranes.

Gene expression analyses of vitellogenin, choriogenin and estrogen receptor subtypes in the livers of male medaka (Oryzias latipes) exposed to equine estrogens.

In the present study, we investigated transcriptional profiles of estrogen-responsive genes, such as vitellogenins (Vtg1 and Vtg2), choriogenins (ChgL and ChgH) and estrogen receptor subtypes (ERα, ERß1, and ERß2), in the liver of male medaka fish (Oryzias latipes) that were exposed to six equine estrogens (1-300 ng l(-1) ) for 3 days. Our quantitative reverse transcription-polymerase chain reaction (RT-PCR) analyses revealed that the expression levels of hepatic Vtg, Chg and ERα genes in male medaka responded to various types and concentrations of equine estrogens. The estrogenic potentials of the tested chemicals were in the order of equilin > 17ß-estradiol > equilenin > 17ß-dihydroequilin > 17ß-dihydroequilenin > 17α-dihydroequilin > 17α-dihydroequilenin, showing the higher estrogenic potential of equilin than that of 17ß-estradiol. Our results also showed that the estrogenicities of 17ß-dihydroequilin and 17ß-dihydroequilenin were more potent than that of 17α-dihydroequilin and 17α-dihydroequilenin. Furthermore, in gene expression analyses of hepatic ER subtypes, observations were made to note that 17ß-estradiol and equilin induced ERα transcription in male medaka, and the ERα transcription level had significantly positive correlations with the expression of Vtg and Chg genes. In contrast, in the same 17ß-estradiol and equilin treatment groups, it was shown that the transcription levels of hepatic ERß1 and/or ERß2 had significantly negative correlations with the expression of Vtg and Chg genes. These results suggested some potential involvement of the ER subtypes in the regulation of Vtg and Chg gene expressions in the liver. This is the first report describing the comprehensive analyses of in vivo estrogenicity of the equine estrogens in male medaka. Copyright © 2016 John Wiley & Sons, Ltd.

An autopsied case of corticobasal degeneration showing severe cerebral atrophy over a protracted disease course of 16 years.

The patient was a 72-year-old Japanese woman. At the age of 57, she started having difficulty performing daily work and developed agraphia. She also exhibited restlessness and loss of interest, and began to speak less. Thereafter, stereotypical behavior, gait disturbance and dysphagia were noted. CT scan demonstrated left-dominant frontal and temporal lobe atrophy. She died at the age of 72, about 16 years after the onset of symptoms. Neuropathologically, the brain weighed 867 g, and showed remarkable cerebral atrophy with degeneration of the white matter, predominantly in the left dorsal frontal lobe and anterior temporal lobe. Microscopically, severe neuronal loss and gliosis with rarefaction were found in the cerebral cortex, and severe destruction of myelin and axons was observed in the cerebral white matter. Moderate neuronal loss with gliosis was also found in the pallidum and substantia nigra. Gallyas-Braak staining and tau immunostaining revealed pretangle neurons, NFTs, ballooned neurons and astrocytic plaques in the cerebral cortex, subcortical nuclei and brainstem, and argyrophilic threads and coiled bodies in the subcortical white matter. Tau isoform-specific immunostaining revealed that most tau-immunoreactive structures were positive for 4-repeat (4R) tau, but some of the NFTs were positive for 3-repeat (3R) tau in the cerebral neocortex. Immunoblotting demonstrated an accumulation of 4R tau in the cerebral cortex and subcortical white matter. The patient was pathologically diagnosed as having corticobasal degeneration. Her long survival course likely accounts for the severe white matter degeneration and accumulation of 3R tau in NFTs.

Endocrine-disrupting potentials of equine estrogens equilin, equilenin, and their metabolites, in the medaka Oryzias latipes: in silico and DNA microarray studies.

Although several previous studies have demonstrated the presence of equine estrogens in the aquatic environment, limited data are currently available on the endocrine-disrupting potentials in fish and the risks they pose to aquatic organisms. To investigate the interactions of major equine estrogens equilin (Eq) and equilenin (Eqn), as well as their metabolites 17α-dihydroequilin, 17ß-dihydroequilin, 17α-dihydroequilenin and 17ß-dihydroequilenin, with the estrogen receptor α (ERα) of medaka (Oryzias latipes), a three-dimensional model of the ligand-binding domain (LBD) of ERα was built in silico, and docking simulations were performed. The docking simulation analysis indicated that the interaction of 17ß-dihydroequilenin with the ERα LBD is the most potent, followed by those of 17α-dihydroequilin and 17ß-dihydroequilin, whereas those of Eq and Eqn were least potent. We further analyzed gene expression profiles in the livers of male medaka exposed to Eq and Eqn. A DNA microarray representing 6000 genes revealed that 24-h exposure to Eq and Eqn (100 ng/L) upregulated the expression of 6 and 34 genes in the livers of males, respectively. Genes upregulated by Eq included the estrogenic biomarker genes vitellogenins and choriogenins, suggesting the estrogenic potential of Eq. In contrast, Eqn exposure upregulated several cancer-related genes, such as mediator complex subunit 16 and RAS oncogene family members, suggesting a carcinogenic potential for Eqn. These results suggest that equine estrogens may have not only endocrine-disrupting potentials via the ERα signaling pathway but also carcinogenic potency in male medaka.

Effects of lithium on growth, maturation, reproduction and gene expression in the nematode Caenorhabditis elegans.

Lithium (Li) has been widely used to treat bipolar disorder, and industrial use of Li has been increasing; thus, environmental pollution and ecological impacts of Li have become a concern. This study was conducted to clarify the potential biological effects of LiCl and Li(2)CO(3) on a nematode, Caenorhabditis elegans as a model system for evaluating soil contaminated with Li. Exposure of C. elegans to LiCl and Li(2)CO(3) decreased growth/maturation and reproduction. The lowest observed effect concentrations for growth, maturation and reproduction were 1250, 313 and 10 000 µm, respectively, for LiCl and 750, 750 and 3000 µm, respectively, for Li(2)CO(3). We also investigated the physiological function of LiCl and LiCO(3) in C. elegans using DNA microarray analysis as an eco-toxicogenomic approach. Among approximately 300 unique genes, including metabolic genes, the exposure to 78 µm LiCl downregulated the expression of 36 cytochrome P450, 16 ABC transporter, 10 glutathione S-transferase, 16 lipid metabolism and two vitellogenin genes. On the other hand, exposure to 375 µm Li(2)CO(3) downregulated the expression of 11 cytochrome P450, 13 ABC transporter, 13 lipid metabolism and one vitellogenin genes. No gene was upregulated by LiCl or Li(2)CO(3). These results suggest that LiCl and Li(2)CO(3) potentially affect the biological and physiological function in C. elegans associated with alteration of the gene expression such as metabolic genes. Our data also provide experimental support for the utility of toxicogenomics by integrating gene expression profiling into a toxicological study of an environmentally important organism such as C. elegans.

[Evaluating the relationship between oral health status in male workers and question item regarding subjective chewing symptoms].

OBJECTIVES: In recent years, the relationship between oral health status and various diseases has been increasingly reported worldwide, leading to a growing understanding of the importance of oral health. This study aimed to investigate the relationship between oral health status, determined after a dental health check-up, and chewing condition, determined via a questionnaire for specific health check-ups with the newly added chewing condition item-When you chew your food, which most accurately describes your chewing-. METHODS: From 20,834 Japanese workers who underwent dental health check-ups and specific health check-ups from April 2018 to March 2019, 4,011 male workers (51.2 ± 6.1 years, range 40-64 years) were extracted for this study. Participants were divided into two groups, the good chewing condition group (GCG) and poor chewing condition group (PCG), according to their responses to a questionnaire regarding chewing conditions. We collected and compared data on subjects' age, smoking status, number of teeth present, number of filled teeth, number of decayed teeth, number of missing teeth with prosthetic treatment, periodontal pockets, gingival bleeding, oral hygiene status, and Eichner's classification. Additionally, to clarify the risk associated with chewing condition, we performed stepwise multiple logistic regression analysis with good chewing condition/poor chewing condition as the dependent variable, and age, smoking status, number of teeth present, number of filled teeth, number of missing teeth with prosthetic treatment, periodontal pockets, oral hygiene status, and Eichner's classification as the independent variables. RESULTS: The result of stepwise multiple logistic regression analyses, the odds ratio (OR) and 95% confidence interval (95% CI) of the factors that remained in the final model were as follows; the number of decayed teeth (OR = 1.18 [95% CI: 1.12-1.24]); periodontal pockets of 3 mm or less vs. 6 mm or more (OR = 1.63 [95% CI: 1.12-2.37]); Eichner's classification A1 vs. A2 (OR = 1.40 [95% CI: 1.08-1.82]), A3 (OR = 1.66 [95% CI: 1.09-2.52]), B1 (OR = 3.38 [95% CI: 2.04-5.61]), B2 (OR = 5.26 [95% CI: 2.46-11.2]), B3 (OR = 4.28 [95% CI: 1.20-15.2]), B4 (OR = 7.59 [95% CI: 1.06-54.5]). CONCLUSIONS: The results of this study suggest a significant association between the newly added "chewing condition" item in the questionnaire for specific health check-ups and oral status; the number of decayed teeth, periodontal pockets, Eichner's classification.

[Association between subjective chewing conditions and dietary habits in Japanese workers].

OBJECTIVES: Previous studies on the elderly have reported that they avoid fiber-rich and hard foods in favor of softer foods as chewing ability declines. However, evidence of worker chewing conditions and dietary habits is scarce. This study aimed to clarify the association between chewing conditions and dietary habits in workers. METHODS: The subjects included 6,703 workers (45.6 ± 10.2 years, range 18-64 years) who underwent medical and dental check-ups from April 2018 to March 2019 with no missing data. We classified workers into two groups, those with good chewing conditions (GCC) and those with poor chewing conditions (PCC) according to their responses to the chewing condition item in our questionnaire. We performed logistic regression analysis using the questionnaire dietary habits item response as the objective variable and chewing conditions as the explanatory variable and adjusting for sex, age, number of present teeth, periodontal pocket depth, oral hygiene status, Body Mass Index, smoking habits, alcohol intake, exercise habits, history of present illness, and stages of transtheoretical model. Additionally, we performed subgroup analysis using a similar framework stratified by sex and age groups (under 40 or older). RESULTS: We found significant negative associations with PCC for the following dietary habits (p < .05): Awareness of balanced diet; Regular consumption of dark green and deep yellow vegetables; Eating slowly and chewing well; Frequent consumption of seaweed and small fish; Having three meals at almost at the same time every day; Having breakfast almost every day; Eating protein dishes with every meal; Frequently consuming dairy products; Finishing eating at least two hours before bedtime. Furthermore, we found significant positive associations with PCC for the following items (p < .05): Having on average two or more bottles/cans of juice or coffee a day; Frequently eating heavy meat dishes; Frequently eating deep-fried foods such as fried dishes and pork cutlets; Frequently having instant or processed foods; Eating two or more kinds of Western or Japanese confectioneries or snacks on average a day; Frequently eating salty food; Regularly having snacks or late-night meals. Additionally, similar trends were also observed following subgroup analysis, regardless of sex or age. CONCLUSIONS: This study suggests that workers with PCC may retain undesirable dietary habits, regardless of sex and age. Therefore, it is necessary to expand dental check-ups in the workplace and to promote dental health guidance that targets all workers.

Association of a third vaccination with antibody levels and side reactions in essential workers: A prospective cohort study.

OBJECTIVE: To evaluate the relationship between the change of titer and adverse events after the third vaccination for COVID-19 among healthcare workers. DESIGN AND SETTING: This was a prospective cohort study, and the follow-up period was from December 2021 to November 2023. PARTICIPANTS: A total of 392 healthcare workers aged over 20 years who worked at the facility and wished to have vaccine antibody titers measured participated in this study. EXPOSURES: A third dose of BNT162b2 COVID-19 vaccine was administered to healthcare workers working at the hospital, and we evaluated the changes in antibody titers before and after the vaccine, as well as adverse reactions after vaccination. MAIN OUTCOMES AND MEASURES: The primary endpoints were adverse reactions within 7 days after the third dose of COVID-19 vaccine and the rate of increase in COVID-19 vaccine antibody titer at 4 weeks. RESULTS: A total of 392 people participated in the study, of whom 358 participants had their antibody titers measured before and after the booster vaccination. The overall IgG geometric mean was 609 U/mL (561-663) before booster vaccination and increased to 18,735 U/mL (17,509-20,049) at 4 weeks after vaccination (p < 0.001). Multivariate analysis showed no statistically significant relationship between the primary endpoints, such as a change in antibody titer due to the presence of fever after vaccination or a change in antibody titer due to swelling at the vaccination site. Factors affecting the rate of increase in antibody titer, evaluated as secondary endpoints, were suggested to be age (1.02 (95 % confidence interval (CI): 1.01-1.03)) and hypertension (0.66 (95 % CI: 0.47-0.93)). CONCLUSIONS AND RELEVANCE: In relation to the booster effect of the third dose of COVID-19 vaccination, there was no statistically significant difference in the presence of fever or use of antipyretic or other drugs.

Effect of curcumin in a mouse model of Pelizaeus-Merzbacher disease.

PLP1 amino acid substitutions cause accumulation of misfolded protein and induce endoplasmic reticulum (ER) stress, causing Pelizaeus-Merzbacher disease (PMD), a hypomyelinating disorder of the central nerve system. Currently no effective therapy is available for PMD. Promoted by its curative effects in other genetic disease models caused by similar molecular mechanisms, we tested if curcumin, a dietary compound, can rescue the lethal phenotype of a PMD mouse model (myelin synthesis deficient, msd). Curcumin was administered orally to myelin synthesis deficit (msd) mice at 180 mg·kg(-1)·day(-1) from the postnatal day 3. We evaluated general and motor status, changes in myelination and apoptosis of oligodendrocytes by neuropathological and biochemical examination, and transcription levels for ER-related molecules. We also examined the pharmacological effect of curcumin in cell culture system. Oral curcumin treatment resulted in 25% longer survival (p<0.01). In addition, oligodendrocytes undergoing apoptosis were reduced in number (p<0.05). However, no apparent improvement in motor function, neurological phenotype, and myelin formation was observed. Curcumin treatment did not change the expression of ER stress markers and subcellular localization of the mutant protein in vitro and/or in vivo. Curcumin partially mitigated the clinical and pathological phenotype of msd mice, although molecular mechanisms underlying this curative effect are yet undetermined. Nonetheless, curcumin may serve as a potential therapeutic compound for PMD caused by PLP1 point mutations.

Does difficulty in chewing induce subjective musculoskeletal symptoms? A case-control study.

OBJECTIVES: Risk factors associated with the development of musculoskeletal disorders and symptoms remain an important issue worldwide. This study aimed to investigate the relationship between oral health problems such as difficulty chewing and the occurrence of stiff neck/shoulders (SN/S) and low back pain (LBP). DESIGN: Case-control study. SETTING AND PARTICIPANTS: This study was conducted from 1 April 2018 to 31 March 2020. The subjects were 77 341 workers among 646 281 workers from several employers in Japan. OUTCOME MEASURES: Participants were asked to evaluate their subjective SN/S and LBP symptoms using a self-administered questionnaire. METHODS: We defined the chewing condition using a questionnaire, and workers who responded with 'I can chew anything' were classified as the good condition group (GCG), and those who responded with 'Sometimes I have difficulty chewing due to problems with the teeth, gums, or bite' or 'I can hardly chew' were classified as the poor condition group (PCG). Setting the year 2018 as the baseline, we performed a logistic regression model using propensity score and inverse probability weighting (IPW) methods and chewing condition groups as explanatory variables and SN/S and LBP as objective variables. RESULTS: The IPW-adjusted logistic regression model showed that the OR of SN/S was approximately 1.25 (95% CI 1.17 to 1.33) times higher in the PCG than that in the GCG (p<0.001). Similarly, the OR of LBP was about 1.37 (95% CI 1.27 to 1.48) times higher in the PCG than that in the GCG in the IPW-adjusted logistic regression model (p<0.001). CONCLUSIONS: Our study suggests that the occurrence of SN/S and LBP symptoms in workers could be predicted depending on the presence of difficulty in chewing. Therefore, oral health and health guidance are gaining importance for the prevention of subjective musculoskeletal symptoms.

Effects of two-year oral health information provision on changes in gingival crevicular fluid in male day-night shift workers.

Background/purpose: Although dental diseases are related to systemic diseases and daily habits and are known to affect the quality of life, little attention has been paid to the importance of oral hygiene in Japanese working-age adults. We, thus, aimed to assess changes in the periodontal condition of working-age adults after providing regular oral health information at their workplace for two years. Materials and methods: Two years of oral health information were provided to workers at four printing companies. The effectiveness thereof was assessed using gingival crevicular fluid (GCF) test results-lactoferrin (Lf), alpha-1-antitrypsin (AT), and aspartate aminotransferase (AST). Questionnaires on health literacy (HL) and dental terminology were also administered to the workers during annual check-ups. Results: The baseline survey participants included were 234 male workers, of which 104 workers followed the same work schedule/day-night shifts and completed the two-year follow-up survey were examined. Providing continuous oral health information with regular updates did not improve participants' self-administered GCF tests (Lf: P = 0.272, AT: P = 0.723, and AST: P = 0.067), and there were no significant changes in HL, perceived symptoms, and use of interdental cleaning tools. Conclusion: Direct oral health education or examination might be necessary for working-age adults to achieve optimum oral conditions, and one-on-one health information provision might not improve their oral condition. A combination of methods to provide oral health promotion must be considered.

Disrupted SOX10 regulation of GJC2 transcription causes Pelizaeus-Merzbacher-like disease.

Mutations in the gap junction protein gamma-2 gene, GJC2, cause a central hypomyelinating disorder; Pelizaeus-Merzbacher-like disease (PMLD; MIM311601). Using a homozygosity mapping and positional candidate gene approach, we identified a homozygous mutation (c.-167A>G) within the GJC2 promoter at a potent SOX10 binding site in a patient with mild PMLD. Functionally, this mutation completely abolished the SOX10 binding and attenuated GJC2 promoter activity. These findings suggest not only that the SOX10-to-GJC2 transcriptional dysregulation is a cause of PMLD, but also that GJC2 may be in part responsible for the central hypomyelination caused by SOX10 mutations.

[Verification of the effects of three percent weight loss at 6 months and application possibility of assessment at 3 months after the specific health guidance for male workers].

OBJECTIVES: This study aimed to evaluate the effects of ≥ 3% body weight loss at 6 months in male workers and verify the application possibility of assessment at 3 months after specific health guidance. METHODS: We recruited 5,031 Japanese male workers (aged 40-64 years) who had undergone health checkups and completed specific health guidance from 2008 to 2017. Participants were divided into three groups according to the percentage of weight reduction (≥ 3%, 0%-3%, ≤ 0%) after 6 months of health guidance. We compared the body weight, waist circumference, and blood pressure at 3- and 6-months with baseline data within and between groups. Similarly, in the medical examination of the next fiscal year (n = 2,889), body weight, waist circumference, blood pressure, triglyceride levels, high-density lipoprotein-cholesterol (HDL-C) levels, plasma glucose levels, and the prevalence and improvement rates of metabolic syndrome (MetS) of specific health checkups were also compared. Furthermore, we calculated the weight reduction target after 3 months to arrive at a ≥ 3% weight reduction for 6 months. Based on the target, we investigated the prevalence and improvement rates of MetS in the next fiscal year. RESULTS: A total of 1,349 (26.8%) participants had a ≥ 3% weight reduction, 2,059 (40.9%) had a < 3% weight reduction, and 1,623 (32.3%) had no weight reduction. At the 6-month follow-up, waist circumference and blood pressure levels had increasingly improved with greater reductions in weight. The following year, body weight, waist circumference, blood pressure, triglyceride levels, HDL-C levels, and plasma glucose levels had improved in the ≥ 3% weight loss group, whereas plasma glucose levels had not improved in the 0-3% weight loss group. Additionally, only triglyceride levels and diastolic blood pressure had improved in the no weight reduction group. In the ≥ 3% weight loss group, the prevalence rate of MetS was the lowest (7.6%), and the improvement rate of MetS was the highest (70.7%). Furthermore, the weight reduction target after 3 months to arrive at a ≥ 3% body weight reduction at 6 months was 2.0% weight reduction. Those who reduced weight ≥ 2.0% had better prevalence and improvement rates of MetS than those who reduced weight < 2.0%. CONCLUSIONS: Our findings indicated that the objective of losing ≥ 3% of body weight at 6 months in male workers after completing specific health guidance was effective and that assessment after 3 months based on setting a 2.0% weight reduction target was applicable.

In vivo and in silico analyses of estrogenic potential of equine estrogens in medaka (Oryzias latipes).

Equine estrogens (EEs) are widely used in hormone replacement therapy pharmaceuticals for postmenopausal women. Previous studies have shown that EEs occur in the aquatic environment; however, the potential estrogenicity and risk of EEs in aquatic organisms, including fish, have yet to be studied in detail. Therefore, we evaluated the estrogenic potential of major EEs, namely equilin (Eq), 17α-dihydroequilin (17α-Eq), 17ß-dihydroequilin (17ß-Eq), equilenin (Eqn), 17α-dihydroequilenin (17α-Eqn), and 17ß-dihydroequilenin (17ß-Eqn), on medaka (Oryzias latipes) using in vivo and in silico assays. Quantitative real-time RT-PCR analyses revealed that expression levels of choriogenin L (ChgL) and choriogenin H (ChgH) in medaka embryos responded to various types and concentrations of EEs in a concentration-dependent manner, whereas transcription levels of vitellogenin 1 were not significantly affected by any of the EEs in the concentration range tested. The order of the in vivo estrogenic potencies of EEs was as follows: 17ß-Eq > Eq > 17ß-Eqn > Eqn > 17α-Eqn > 17α-Eq. Additionally, the 50% effective concentrations (EC50) of 17ß-Eq was lower than that of 17ß-estradiol. We also investigated the interaction potential of EEs with medaka estrogen receptor (ER) subtypes in silico using a three-dimensional model of the ligand-binding domain (LBD) for each ER and docking simulations. All six EEs were found to interact with the LBDs of ERα, ERß1, and ERß2. The order of the in silico interaction potentials of EEs with each ER LBD was as follows: 17ß-Eq > 17α-Eq > Eq > 17ß-Eqn > 17α-Eqn > Eqn. Furthermore, we identified the key amino acids that interact with EEs in each ER LBD; our findings suggest that amino acids and/or their hydrogen bonding may be responsible for the ligand-specific interactions with each ER. This study is the first to comprehensively analyze the estrogenic potential of EEs in medaka both in vivo and in silico.

Construction of a (18)F-FDG PET normative database of Japanese healthy elderly subjects and its application to demented and mild cognitive impairment patients.

OBJECTIVE: To construct a (18)F-FDG PET normative database of Japanese healthy elderly subjects and to apply it to demented and mild cognitive impairment (MCI) patients. METHODS: Seventy-seven Japanese normal volunteers from 41 to 84 years of age (36 males and 41 females) who underwent clinical, neuropsychological, and MRI examinations were selected. In these subjects, (18)F-FDG PET/CT scans were performed, (18)F-FDG PET images were analyzed using the 3D-SSP program, and a normative database for cerebral glucose metabolism was constructed. Then, (18)F-FDG PET images from 14 demented and MCI patients were evaluated based on the normative database. RESULTS: The 77 healthy elderly subjects were divided into three groups according to their age. In these subjects, the difference in glucose metabolism between males and females was minimal in contrast, glucose metabolism showed a weak reciprocal correlation with aging in several cerebral regions. The 3D-SSP images of 14 demented and MCI patients based on the age-matched (18)F-FDG PET normative database showed decreased patterns of glucose metabolism similar to those of previous studies on dementia diseases and MCI. CONCLUSIONS: An age-matched normative database can be applied to the evaluation of single subjects, and the application of a mixed database of males and females is viable. Normative databases are useful for detecting dementia diseases and their MCI.

Diffuse occipital hypometabolism on [18 F]-FDG PET scans in patients with idiopathic REM sleep behavior disorder: prodromal dementia with Lewy bodies?

BACKGROUND: Previous longitudinal studies have revealed that specific patterns on [(18) F]-fluoro-d-glucose (FDG) positron emission tomography (PET) scans in patients with amnesic mild cognitive impairment can predict Alzheimer's disease (AD). However, the significance of particular patterns on [(18) F]-FDG PET scans in prodromal patients with dementia with Lewy bodies (DLB) remains unclear. METHODS: Based on the prevailing evidence that rapid eye movement (REM) sleep behavior disorder (RBD) often precedes the onset of DLB, [(18) F]-FDG PET scans of nine non-demented patients reporting recurrent nocturnal dream-enactment behavior in our memory clinic were compared with the normative database using three-dimensional stereotactic surface projection (3D-SSP) images. All patients underwent clinical and neuropsychological examinations as well as cardiac [(123) I]-metaiodobenzylguanidine ([(123) I]-MIBG) scintigraphy. RESULTS: Four patients were found to have diffuse areas of reduced cerebral metabolic rate of glucose (CMRglc), predominantly in the occipital lobe, which is the preferentially affected region in DLB patients. In contrast, five patients showed no such occipital hypometabolism; instead, these five patients showed hypometabolism in the left anterior cingulate gyrus (Broadmann area (BA) 24), right frontal lobe (BA 32) and right anterior temporal lobe (BA 38), which are the preferentially affected regions in Parkinson's disease rather than DLB. The extent of the reduction in CMRglc in the left occipital lobe was correlated with scores on the Bender Gestalt Test, which reflects visuospatial ability, but not with global cognitive measures. All patients showed reduced cardiac [(123) I]-MIBG levels, consistent with underlying Lewy body disease. CONCLUSION: These variations in [(18) F]-FDG PET scans raise the possibility that the specific pattern of CMRglc reduction may predict developing DLB in patients with idiopathic RBD. Further follow-up studies are needed, particularly on patients with diffuse occipital hypometabolism.

Detection of early amnestic mild cognitive impairment without significantly objective memory impairment: a case-controlled study.

BACKGROUND: We encountered eight early amnestic mild cognitive impairment (aMCI) patients (early MCI group) who did not fulfill the diagnostic criteria for aMCI. We compared the scores of neuropsychological examinations as well as the cerebral metabolic rate for glucose consumption (CMRglc) decrease on (18)F-FDG PET examination between the early MCI group and 10 aMCI patients (MCI group) or six normal elderly subjects (normal group), to examine whether the current diagnostic criteria can detect early-stage aMCI. METHODS: The three groups underwent Mini-Mental State Examination (MMSE), Wechsler Adult Intelligence Scale - Third Edition (WAIS-III), Wechsler Memory Scale Revised (WMS-R), magnetic resonance imaging and 18F-fluorodeoxyglucose positron emission tomography ((18)F-FDG PET) examinations. RESULTS: The early MCI group did not show significant memory impairment of 1.0 SD or other cognitive dysfunctions on neuropsychological examinations, and did not fulfill the diagnostic criteria of aMCI. With one-way anova and Tukey's HSD post-hoc test, the early MCI group showed the highest scores for WAIS-III, whereas the MCI group showed the lowest scores for WMS-R, although there were no significant differences between the early MCI and normal groups. In order to show a discrepancy in scores between WAIS-III and WMS-R, we subtracted the scores of WMS-R from WAIS-III. Consequently, the normal group showed significantly smaller differences in scores than the other groups, although there were no significant differences between the early MCI and MCI groups. (18)F-FDG PET recognized a CMRglc decrease in the posterior cingulate gyrus and/or part of the parietotemporal area in both the MCI and early MCI groups, of which the extent and magnitude were weaker in the early MCI group. The normal group did not show a significant CMRglc. CONCLUSIONS: The early MCI group should be included in aMCI not only based on the discrepancy between intelligence and memory scores, but also based on the (18)F-FDG PET findings. The combination of these examinations would make it possible to diagnose early-stage aMCI.

Abnormal localization of leucine-rich repeat kinase 2 to the endosomal-lysosomal compartment in lewy body disease.

Missense mutations in the leucine-rich repeat kinase 2 (LRRK2) gene are the most common causes of both familial and sporadic forms of Parkinson disease and are also associated with diverse pathological alterations. The mechanisms whereby LRRK2 mutations cause these pathological phenotypes are unknown. We used immunohistochemistry with 3 distinct anti-LRRK2 antibodies to characterize the expression of LRRK2 in the brains of 21 subjects with various neurodegenerative disorders and 7 controls. The immunoreactivity of LRRK2 was localized in a subset of brainstem-type Lewy bodies (LBs) but not in cortical-type LBs, tau-positive inclusions, or TAR-DNA-binding protein-43-positive inclusions. The immunoreactivity of LRRK2 frequently appeared as enlarged granules or vacuoles within neurons of affected brain regions, including the substantia nigra, amygdala, and entorhinal cortex in patients with Parkinson disease or dementia with LBs. The volumes of LRRK2-positive granular structures in neurons of the entorhinal cortex were significantly increased in dementia with LBs brains compared with age-matched control brains (p < 0.05). Double immunolabeling demonstrated that these LRRK2-positive granular structures frequently colocalized with the late-endosomal marker Rab7B and occasionally with the lysosomal marker, the lysosomal-associated membrane protein 2. These results suggest that LRRK2 normally localizes to the endosomal-lysosomal compartment within morphologically altered neurons in neurodegenerative diseases, particularly in the brains of patients with LB diseases.