Showing papers by "Gifu University of Medical Science published in 2021"

Acylative Desymmetrization of Cyclic meso-1,3-Diols by Chiral DMAP Derivatives

Abstract: An efficient enantioselective acylative desymmetrization of cyclic meso-1,3-diols was developed by using a chiral DMAP derivative 1e having a 1,1′-binaphthyl unit. The reactions required only 0.5 m...

Social engagement, self‐efficacy, and posttraumatic stress symptoms across 6 months of psychotherapy

Abstract: OBJECTIVE The current study was conducted in a naturalistic treatment setting to examine whether and how perceptions about social engagement, trauma coping self-efficacy, and posttraumatic stress symptoms (PTS) influence one another across 6 months of psychotherapy for trauma survivors. METHOD The sample included 183 clients who reported exposure to traumatic events and significant PTS (PCL-5 ≥ 33). Participants (Mage = 37.8, 53.6% female) completed surveys at intake, 3 months, and 6 months into treatment. A cross-lagged panel analysis was used to test the relationships among perceived social engagement, coping self-efficacy, and PTS across three assessment points. RESULTS PTS at 3-months was a mediator in the relationship between intake perceived social engagement and 6-month coping self-efficacy and between intake perceived social engagement and 6-month perceived social engagement. CONCLUSIONS PTS several months into treatment may serve as a mechanism between intake perceived social engagement and functional outcomes such as coping self-efficacy.

Heterogeneous induction of an invasive phenotype in prostate cancer cells by coculturing with patient-derived fibroblasts.

Abstract: Prostate cancer (PCa) cells frequently invade the surrounding stroma, leading to heterogeneous formation of structural atypia. The surrounding stroma contains multiple functionally diverse populations of fibroblasts that trigger numerous changes in PCa cells including motility. Thus, we hypothesized that direct or indirect contact of PCa cells with fibroblasts determines an invasive phenotype in PCa cells. We investigated the effects of 10 different patient-derived fibroblast lines on the three-dimensional (3D) morphogenesis of PCa cells growing on a viscous substrate in vitro. When grown alone, all 10 patient-derived fibroblast lines clumped on the viscous substrate, whereas the human androgen-sensitive PCa cell line LNCaP did not. Cocultures of LNCaP cells with seven of the patient-derived fibroblast lines (PrSC, pcPrF-M5, pcPrF-M7, pcPrF-M23, pcPrF-M24, pcPrF-M28, and pcPrF-M31) formed a thick fibroblast layer that resembled human prostate stromal structures. In contrast, cocultures of LNCaP cells with the remaining three fibroblast lines (NPF-M13, pcPrF-M10, and pcPrF-M26) did not form a thick fibroblast layer. Of the seven fibroblast lines that caused thick layer formation, four patient-derived fibroblast lines (PrSC, pcPrF-M5, pcPrF-M28, and pcPrF-M31) induced an invasive phenotype in LNCaP cells with a cord-like infiltrating growth pattern, whereas the other three fibroblast lines (pcPrF-M7, pcPrF-M23, and pcPrF-M24) induced no or a very weak invasive phenotype. Using cell culture inserts, none of the four patient-derived fibroblast lines that induced an invasive phenotype (PrSC, pcPrF-M5, pcPrF-M28, and pcPrF-M31) affected CDH1 mRNA expression in LNCaP cells; yet, two patient-derived fibroblast lines (pcPrF-M5 and pcPrF-M28) increased CDH2 mRNA expression in LNCaP cells, whereas the other two fibroblast lines (PrSC and pcPrF-M31) did not. These results suggest that the existence of multiple functionally diverse populations of fibroblasts in PCa tissue may be responsible for the diversity in PCa cell invasion, leading to heterogeneous formation of structural atypia.

The Oxidation of Equol by Tyrosinase Produces a Unique Di-ortho-Quinone: Possible Implications for Melanocyte Toxicity

Abstract: Equol (7-hydroxy-3-(4'-hydroxyphenyl)-chroman, EQ), one of the major intestinally derived metabolites of daidzein, the principal isoflavane found in soybeans and most soy foods, has recently attracted increased interest as a health-beneficial compound for estrogen-dependent diseases. However, based on its structure with two p-substituted phenols, this study aimed to examine whether EQ is a substrate for tyrosinase and whether it produces o-quinone metabolites that are highly cytotoxic to melanocyte. First, the tyrosinase-catalyzed oxidation of EQ was performed, which yielded three EQ-quinones. They were identified after being reduced to their corresponding catechols with NaBH4 or L-ascorbic acid. The binding of the EQ-quinones to N-acetyl-L-cysteine (NAC), glutathione (GSH), and bovine serum albumin via their cysteine residues was then examined. NAC and GSH afforded two mono-adducts and one di-adduct, which were identified by NMR and MS analysis. It was also found that EQ was oxidized to EQ-di-quinone in cells expressing human tyrosinase. Finally, it was confirmed that the EQ-oligomer, the EQ oxidation product, exerted potent pro-oxidant activity by oxidizing GSH to the oxidized GSSG and concomitantly producing H2O2. These results suggest that EQ-quinones could be cytotoxic to melanocytes due to their binding to cellular proteins.

In vitro metabolic profiles of adamantyl positional isomers of synthetic cannabinoids

Abstract: Illegal use of synthetic cannabinoids (SCs) is a serious problem worldwide. Legal regulation of SCs requires fundamental analytical studies regarding the differentiation of potential structural isomers. Accumulation of SC metabolic profiles is also essential for forensic investigation because SCs are immediately metabolized after intake. Thus, we investigated the in vitro metabolism of N-adamantyl-1-(tetrahydropyran-4-ylmethyl)-1H-indazole-3-carboxamide isomers (ATHs) using human liver microsomes (HLMs). Moreover, we validated the applicability of the isomeric differentiation by investigation of N-adamantyl-1-(4-fluorobenzyl)-1H-indazole-3-carboxamide isomers (AFUs). Metabolites were collected at designated time points during the incubation period with HLMs for up to 180 min. The structures of the metabolites were annotated on the basis of mass spectroscopic evidence obtained by liquid chromatography–ion trap–time of flight mass spectrometry. The secondary stage mass (MS2) spectra obtained from the protonated molecules revealed a clear difference in both ATHs and their major metabolites because of the stability of the adamantyl (AD) cation. In HLMs, ATHs were quickly metabolized, and hydroxylation of the AD ring was deduced as the major metabolic pathway. The major metabolites of ATH 1 and ATH 2 after 180 min showed dihydroxylation and monohydroxylation of the AD ring. The AFUs showed analytical and metabolic profiles similar to those of the ATHs described above. We characterized the metabolism of ATHs for the first time and discriminated between the two isomers by mass spectrometric analysis of either the parent compounds or their major metabolites. Our investigation of AFUs also demonstrated a useful method for distinguishing between AD isomers.

Intimate Adhesion Is Essential for the Pathogen-Specific Inflammatory and Immune Responses in the Gut of Mice Infected with Citrobacter rodentium.

Abstract: Citrobacter rodentium is a murine pathogenic bacterium that adheres to intestinal epithelial cells, resulting in loss of microvilli and pedestal formation, and alters multiple cellular processes, including actin dynamics. Translocated intimin receptor (Tir), one of its virulence factors, functions as receptor for intimin, a bacterial adhesin, thereby mediating bacterial adhesion to epithelial cells. Although robust immune responses are induced to eliminate pathogenic bacteria in the host, they are suppressed against harmless commensal bacteria. The mechanism(s) underlying such a differentiation remains unclear. This study sought to determine the roles of intimate adhesion in the induction of specific immune responses upon C. rodentium infection. To this end, microbiota-depleted mice were infected with the Tir-F strain expressing full-length Tir or mutant strains expressing the C-terminal truncated Tir that is defective in intimin binding and host cell actin polymerization. There were no differences in the colonization kinetics and Abs responses against C. rodentium LPS among the strains, whereas Abs against the virulence factors were only produced on Tir-F infection. Although there were no differences in the virulence factors mRNA expression levels, colonic hyperplasia, and bacterial translocation to the systemic organs irrespective of the strain, adhesion to colonic epithelial cells was reduced in the mutant strain-infected mice. Furthermore, transcriptomic analysis indicated that robust inflammatory and immune responses were only induced in the Tir-F-infected group and were suppressed in the mutant-infected groups. Taken together, these findings suggest that Tir-mediated intimate adhesion induces inflammatory and immune responses, resulting in the induction of virulence factor-specific Abs.

Alginate-coated activated charcoal enhances fecal excretion of 2,3,7,8-tetrachlorodibenzo-p-dioxin in mice, with fewer side effects than uncoated one.

Abstract: Activated charcoal (AC) is a potential candidate antidote against dioxins. However, it is difficult to take AC as a supplement on a daily basis, because its long-term ingestion causes side effects such as constipation and deficiency of fat-soluble essential nutrients and hypocholesterolemia. Alginate-coated AC, termed Health Carbon (HC), was developed to decrease the side effects of AC, but its pharmacological effects, including side effects, remains unclear. Here, we show that HC enhanced fecal excretion of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) and decreased some side effects of unmodified AC, such as hypocholesterolemia, in male mice. Basal diet mixed with HC or unmodified AC at various concentrations was fed to mice for 16 days following a single intraperitoneal administration of [3H]TCDD. Both HC and unmodified AC at 3% or more significantly increased fecal excretion of [3H]TCDD in comparison with the control basal diet. Consistent with this, [3H]TCDD radioactivity in the liver-a major TCDD storage organ-was markedly decreased by HC at concentrations of 3% and 10%. In an examination of potential side effects, unmodified AC at 10% or more caused significant body weight reduction and at 20% caused significant hypocholesterolemia. In contrast, HC caused weight gain reduction only at a concentration of 20%, and there was no evidence of hypocholesterolemia at any dietary HC concentration. HC not only retains the ability of AC to enhance fecal excretion of TCDD but also reduces some of the side effects of AC.

Quantification of ustalic acid, a chemotaxonomic marker, in Tricholoma ustale using liquid chromatography–mass spectrometry

Abstract: The development of methods for the detection and qualification of toxic substances in mushrooms is a rapidly growing research area in forensic toxicology. This study aimed to determine liquid chromatographic and mass spectrometric conditions applicable to the analysis of ustalic acid (UA) in Tricholoma ustale. We used HPLC coupled with single quadrupole mass spectrometry (Q-MS) with electrospray ionization in its negative ion mode to analyze UA. We performed HPLC separations on a C18 reversed-phase column with gradient elution using mobile phases containing water, acetonitrile, and formic acid. The MS showed that UA formed the deprotonated molecular ion [M–H]− at m/z 337, which was sufficient for the quantitative analysis of the compound. The average recovery rates of UA from four edible mushrooms (Flammulina velutipes, Pleurotus eryngii, Lentinula edodes, and Grifola frondosa) to which 10.0 μg/g of UA was added were 108%, 104%, 108%, and 107%, respectively, and the relative standard deviation values ranged from 4.1 to 6.4%. Quantitative analysis of UA in three systematically collected individual mushrooms of T. ustale revealed 41.9–155.7 ppm in each dry material. We also explored the fragmentation behaviors of UA in triple quadrupole mass spectrometry and the proposed structures for the product ions. The data suggest that conventional Q-MS with authenticated UA would be able to identify this compound in T. ustale when used for the immediate inspection of incidences of poisoning. Confirmation of the presence of UA in T. ustale would ultimately allow for the chemotaxonomic discrimination of Tricholoma species.

Effect of computed tomography value error on dose calculation in adaptive radiotherapy with Elekta X-ray volume imaging cone beam computed tomography.

Abstract: Purpose We evaluated the effect of changing the scan mode of the Elekta X-ray volume imaging cone beam computed tomography (CBCT) on the accuracy of dose calculation, which may be affected by computed tomography (CT) value errors in three dimensions. Methods We used the electron density phantom and measured the CT values in three dimensions. CT values were compared with planning computed tomography (pCT) values for various materials. The evaluated scan modes were for head and neck (S-scan), chest (M-scan), and pelvis (L-scan) with various collimators and filter systems. To evaluate the effects of the CT value error of the CBCT on dose error, Monte Carlo calculations of dosimetry were performed using pCT and CBCT images. Results The L-scan had a CT value error of approximately 800 HU at the isocenter compared with the pCT. Furthermore, inhomogeneity in the longitudinal CT value profile was observed in the bone material. The dose error for ±100 HU difference in CT values for the S-scan and M-scan was within ±2%. The center of the L-scan had a CT error of approximately 800 HU and a dose error of approximately 6%. The dose error of the L-scan occurred in the beam path in the case of both single field and two parallel opposed fields, and the maximum error occurred at the center of the phantom in the case of both the 4-field box and single-arc techniques. Conclusions We demonstrated the three-dimensional CT value characteristics of the CBCT by evaluating the CT value error obtained under various imaging conditions. It was found that the L-scan is considerably affected by not having a unique bowtie filter, and the S-scan without the bowtie filter causes CT value errors in the longitudinal direction. Moreover, the CBCT dose errors for the 4-field box and single-arc irradiation techniques converge to the isocenter.

3-Hydroxykynurenine Regulates Lipopolysaccharide-Stimulated IL-6 Production and Protects against Endotoxic Shock in Mice.

Abstract: Despite advances in our understanding of endotoxic shock, novel therapeutic interventions that can reduce the burden of sepsis remain elusive. Current treatment options are limited, and it is only through refinements in the ways that we deliver supportive care that mortality has fallen over the years. In this study, the role of kynurenine 3-monooxygenase (KMO) in immune regulation was examined in LPS-induced endotoxemia using KMO-/- and KMO+/+ mice treated with the KMO inhibitor Ro61-8048. We showed that LPS-induced or cecal ligation and puncture-induced mortality and hepatic IL-6 production increased in the absence of KMO, possibly involving increased activating transcription factor 4 (ATF4) signaling in hepatic macrophages. Moreover, treatment of septic mice with 3-hydroxykynurenine reduced mortality rates and inflammatory responses regardless of the presence or absence of KMO. According to our results, the administration of 3-hydroxykynurenine as part of the treatment approach for sepsis or as an adjuvant therapy might reduce the overproduction of IL-6, which is responsible for severe endotoxemia, and ultimately improve the survival rates of patients with sepsis.

A Study for Statistical Machine Learning to Classify Radial Motions of the Elderly

Abstract: Human visual information in peripheral and foveal vision fields is processed along dorsal and ventral streams in the brain, respectively. It has been also reported that the motion sickness is induced by visual approaches especially in the peripheral vision. In general, difference in the radial motion has been found between the peripheral and the foveal vision. In this study, we measured eye movements during the peripheral or foveal vision of 3D video clips and conducted statistical machine learning to classify sample data into two types of radial motion processes. We measured the eye movements of six healthy elderly people while viewing 3D video clips. We extracted about 2,300 time series data of these eye movements for 1, 2, 3, 4, 5, 10, and 20 seconds, respectively. These time series were used as a data set, and statistical machine learning was performed for the classification. As a result, the accuracy of the classification increased with the sample size. In addition, a comparison of the correct rate of the classification between peripheral and foveal vision showed that the rate was higher for the foveal vision.

Real-Time Estimation of Eye Movement Condition Using a Deep Learning Model

Abstract: In this study, we conducted a basic investigation involving the discrimination of eye movement condition (peripheral and central vision) using deep learning techniques. The subjects were 6 males aged 21–23 years. They watched two three-minute videos for central vision and peripheral vision in a random order for a total of eight sessions (four sessions each). The subjects wore an eye movement measurement device, and their eye movements (viewing angles) during the viewing of each video were continuously. From the time series data for eye movement, with four different lengths (0.5 s, 1 s, 2 s, 3 s) and shift length of 0.5 s, short time series data for each 3 min was obtained in sets of 350, and the data were utilized for deep learning and its evaluation. For the deep learning model, input nodes according to data length were placed in the input layer. For the middle layer, seven to eight units were put in place that brought together the one-dimensional convolution layer, the batch-normalization layer, normalized linear function, and the max-pooling layer. The output layer consisted of the fully-connected layer, sigmoid function, and multi-class cross-entropy. As a result, the accuracy of the discrimination was improved as the data length increased, and it was possible to determine the condition with an accuracy of over 90% if the eye movement data was at least one second.

ETV6-related thrombocytopenia associated with a transient decrease in von Willebrand factor.

Abstract: ETV6-related thrombocytopenia is an autosomal dominant thrombocytopenia, characterized by a bleeding tendency and predisposition to hematological malignancies. The similarity in symptoms makes differentiating immune and congenital thrombocytopenia challenging. We report a 5-year-old girl who presented with chronic thrombocytopenia associated with repetitive and long-lasting epistaxis, leading to blood transfusion for severe anemia. Blood tests showed thrombocytopenia (52 × 103/µL) with normal-sized platelets and transiently low von Willebrand factor (VWF) levels (VWF:RCo 13%, VWF:Ag 50%); therefore, von Willebrand disease type 2 was initially suspected. Repetition of the blood tests revealed normal levels of VWF. Exome and Sanger sequencing identified a germline ETV6 heterozygous variant, c.641C > T:p.(P214L). No additional pathogenic variants were found, including VWF, in the gene panel testing of the 53 known target causative genes for thrombocytopenia. High-throughput exome sequencing for chronic thrombocytopenia can be utilized to differentially diagnose ETV6-related thrombocytopenia from chronic/intractable immune thrombocytopenia and to effectively monitor malignancy.

Artificial Intelligence for Medical Imaging and Hygiene

Abstract: Artificial Intelligence (AI) is commonly defined as machine intelligence that is programmed to think like humans and emulate their behavior or simply the technology of developing AI. People receive great benefits from services based on AI technology without even realizing it. One of the technologies that underlie AI is machine learning, and deep learning is a new type of machine learning. Mathematical models of deep learning usually employ a neural network model that imitates the human brain neural network, and a deep neural network has some middle layers to solve the highly difficult issues. A Convolutional Neural Network (CNN), which is a type of deep learning network, can analyze an image directly. In the field of medical imaging and diagnosis, deep learning is already being used to solve numerous difficult issues. Systems have been developed using CNNs and have achieved practical medical application. This chapter focuses in detail on the relationship between deep learning, medical imaging, and hygiene. In particular, it contains an explanation and technical summary of machine learning, the application of deep learning in medical imaging, and the association of the Coronavirus disease 2019 (COVID-19) infection with hygiene and deep learning.

[Construction and Analysis of a Digital Mammography Database: Analysis of Breast Compression Pressures at Four Facilities].

Abstract: We analyzed the compression pressures in 2772 mammography images of 807 patients acquired by digital mammography equipment at four facilities. The analysis included the average compression pressure at all facilities, difference in compression pressure at each facility, differences between the pressures used by radiological technologists in the same facility, and difference attributed to the breast structure. We also analyzed the effects of the compression pressure on the breast thickness and mean glandular dose (MGD) at each facility. The median values of the compression pressure and breast thickness for the 2772 images at all facilities were 86.5 N and 43 mm, respectively. The compression pressures differed among the institutions. The maximum difference in the median pressures among the four facilities was 38.6 N, while the difference in the breast thickness was 6 mm. The radiological technologists working at the same facility used almost the same compression pressure. However, differences between the compression pressures used by different radiological technologists were observed. The compression pressure in a dense breast was smaller than that in a non-dense breast. The difference in the compression pressure affected the breast thickness and MGD. The results of this analysis could be utilized for an optimal imaging in future digital mammography.

MYH9 disorder: Identification and a novel mutation in patients with macrothrombocytopenia.

Abstract: The diagnosis of MYH9 disorder is guided by recognizing granulocyte Dohle body-like inclusion bodies and large/giant platelets in the peripheral blood smear. Immunofluorescence study of nonmuscle myosin heavy chain IIA is a sensitive screening method for diagnosis of MYH9 disorder. The diagnosis can then be confirmed by genetic analysis. A total of 67 patients with macrothrombocytopenia were included, of which 11 patients (16%), aged 4 months to 22 years, were ultimately diagnosed with MYH9 disorder. One novel mutation in exon 30 at c.4338T>C (p.F1446A) was detected. This mutation was associated with nonhematologic manifestations presenting in late adolescence with cataracts, hearing loss, and hematuria.

Myh9 R702C is associated with erythroid abnormality with splenomegaly in mice.

Abstract: MYH9 disorders are characterized by giant platelets, thrombocytopenia, and Dohle body-like cytoplasmic inclusion bodies in granulocytes. However, whether these disorders cause any changes in erythroid cells has yet to be determined. This study analyzed the influence of Myh9 R702C, as one of the most commonly detected MYH9 disorders, on erythroid cells in a mouse model. Knock-in mice expressing Myh9 R702C mutation either systemically or specific to hematological cells (R702C and R702C vav1 mice, respectively) were used in this study. Both displayed lower hemoglobin and higher erythropoietin levels than wild-type (WT) mice, along with significant splenomegaly. Flow cytometric analysis revealed erythroblasts present at a higher rate than WT mice in the spleen. However, no obvious abnormalities were seen in erythroid differentiation from megakaryocyte/erythroid progenitor to erythrocyte. Cell culture assay by fetal liver and colony assay also showed normal progression of erythroid differentiation from erythroid burst-forming unit to red blood cell. In conclusion, R702C and R702C vav1 mice displayed erythroid abnormality with splenomegaly. However, erythroid differentiation showed no obvious abnormality. Further research is required to elucidate the underlying mechanisms.

The Fungal Metabolite (+)-Terrein Abrogates Ovariectomy-Induced Bone Loss and Receptor Activator of Nuclear Factor-κB Ligand–Induced Osteoclastogenesis by Suppressing Protein Kinase-C α/βII Phosphorylation

Abstract: Osteoporosis is a common disease characterized by a systemic impairment of bone mass and microarchitecture that results in fragility fractures. Severe bone loss due to osteoporosis triggers pathological fractures and consequently decreases the daily life activity and quality of life. Therefore, prevention of osteoporosis has become an important issue to be addressed. We have reported that the fungal secondary metabolite (+)-terrein (TER), a natural compound derived from Aspergillus terreus, has shown receptor activator of nuclear factor-κB ligand (RANKL)-induced osteoclast differentiation by suppressing nuclear factor of activated T-cell 1 (NFATc1) expression, a master regulator of osteoclastogenesis. TER has been shown to possess extensive biological and pharmacological benefits; however, its effects on bone metabolism remain unclear. In this study, we investigated the effects of TER on the femoral bone metabolism using a mouse-ovariectomized osteoporosis model (OVX mice) and then on RANKL signal transduction using mouse bone marrow macrophages (mBMMs). In vivo administration of TER significantly improved bone density, bone mass, and trabecular number in OVX mice (p < 0.01). In addition, TER suppressed TRAP and cathepsin-K expression in the tissue sections of OVX mice (p < 0.01). In an in vitro study, TER suppressed RANKL-induced phosphorylation of PKCα/βII, which is involved in the expression of NFATc1 (p < 0.05). The PKC inhibitor, GF109203X, also inhibited RANKL-induced osteoclastogenesis in mBMMs as well as TER. In addition, TER suppressed the expression of osteoclastogenesis-related genes, such as Ocstamp, Dcstamp, Calcr, Atp6v0d2, Oscar, and Itgb3 (p < 0.01). These results provide promising evidence for the potential therapeutic application of TER as a novel treatment compound against osteoporosis.