Summary of “A Basic Study on Bodhisattva-caryā in Buddhâvataṃsaka-sūtra”

departmental bulletin pape

Bioconversion of chicken feather into amino acids and keratinase production by mesophilic Chryseobacterium proteolyticum and Pseudomonas aeruginosa isolated from municipal waste dumpsites

Chicken feathers are by-products of poultry processing which are generated in large amount because of the global growing demand for poultry meats. They have high contents of crude proteins in the form of keratin which could be valorized into digestible products. Keratinases are classified as a specific collection of proteolytic enzymes that have the ability for the degradation of recalcitrant keratinous substrates. Isolation and characterization of these enzymes from various microbial producers are gaining prominence in recent years due to their industrial and biotechnological application potentials. For this research, the collection of soil samples was done as well as the isolation of bacteria and the screening for keratinolytic activity. 16S rDNA sequencing and phylogenetic analysis were used to identify the isolates with efficient chicken feathers degrading capacity. Optimum conditions for the fermentation prcocess was enhanced for the production of keratinase. The fermentation broth was also analysed for various amino acids of protein, and the biochemical properties of the enzymes were likewise determined. Twenty two (22) bacteria were isolated from the soil samples, and 18 out of the 22 isolates showed proteolytic activity on solid media with diameters of halo zone that ranged from 5 ± 0.71 mm for isolate coded as PSS-03 to 25 ± 1.41 mm for isolate coded as PSS-06. Intact chicken feathers were degraded by proteolytic bacterial isolates in variable degree that ranged from 24percent for PSS-10 and 81percent for DSS-02. Extracellular keratinase production recorded for the isolates ranged from 63.63 ± 4.14 U/mL for PSS-10 to 693.63 ± 62.99 U/mL for DSS-02. Based on 16S rDNA sequence and phylogenetic analysis, the 2 isolates with remarkable keratinolytic activity coded as DSS-02 and PSS-14 were identified as Chryseobacterium proteolyticum FGNn and Pseudomonas aeruginosa GNFx. C. proteolyticum showed the maximum keratinase production of 756.36 U/mL after 72 h of incubation at optimized fermentation conditions which involved initial medium pH (4), incubation temperature (30 oC), inoculum size (2percent; v/v), and chicken feathers (1.5percent; w/v). Similarly, P. aeruginosa optimally produced keratinase (1055.45 U/mL) after 96 h of incubation at optimized fermentation conditions that involved initial medium pH (7-8), incubation temperature (30 oC), inoculum size (5percent; v/v), and chicken feathers (2.5percent; w/v). Furthermore, feather hydrolysate from C. proteolyticum FGNn had relatively higher abundance (>1.5g/100g sample) of arginine (1.85), serine (1.63), glycine (1.9) and lysine (1.62); while P. aeruginosa GNFx feather hydrolysate showed high abundance of arginine, serine, aspartic acid, glutamic acid, glycine, alanine, valine, and leucine with respective concentration of 2.06, 1.67, 2.39, 3.05, 1.87, 1.73, 1.56 and 1.65 (g/100g sample). The results showed that keratinases from the two bacterial isolates were optimally active at pH 8, and temperature of 50 oC for FGNn keratinase and 50-60 oC for GNFx keratinase. The enzymes displayed remarkable pH stability. Keratinase from C. proteolyticum was catalytically inhibited by EDTA and 1,10-phenanthroline but not affected by PMSF; while P. aeruginosa keratinase was not significantly affected by those class of protease inhibitors. Adiitionally, FGNn keratinase demonstrated high residual activity of 90percent, 103percent, 101percent, 110percent, 130, and 105percent in the presence of DTT, hydrogen peroxides, acetonitrile, triton X-100, tween-80 and SDS, respectively. Similarly, catalytic efficiency of GNFx keratinase was promoted in the presence of hydrogen peroxides (119percent), triton X-100 (140percent), tween-80 (150percent) and SDS (147percent) compared to the control. Furthermore, the keratinases from the both bacterial isolates exhibited catalytic efficiency enhancement and remarkable structural stability in the presence of laundry detergents tested. The findings from the study suggest the application potentials of the isolates for the bioconversion of recalcitrant keratinous wastes into digestible and quality protein hydrolysates. The properties of these microbial keratinases indicate that they may be exploited for various biotechnological and industrial processes especially in the formulation of detergents.Thesis (MSc) -- Faculty of Science and Agriculture, 202

Bioconversion of chicken feather into amino acids and keratinase production by mesophilic Chryseobacterium proteolyticum and Pseudomonas aeruginosa isolated from municipal waste dumpsites

Chicken feathers are by-products of poultry processing which are generated in large amount because of the global growing demand for poultry meats. They have high contents of crude proteins in the form of keratin which could be valorized into digestible products. Keratinases are classified as a specific collection of proteolytic enzymes that have the ability for the degradation of recalcitrant keratinous substrates. Isolation and characterization of these enzymes from various microbial producers are gaining prominence in recent years due to their industrial and biotechnological application potentials. For this research, the collection of soil samples was done as well as the isolation of bacteria and the screening for keratinolytic activity. 16S rDNA sequencing and phylogenetic analysis were used to identify the isolates with efficient chicken feathers degrading capacity. Optimum conditions for the fermentation prcocess was enhanced for the production of keratinase. The fermentation broth was also analysed for various amino acids of protein, and the biochemical properties of the enzymes were likewise determined. Twenty two (22) bacteria were isolated from the soil samples, and 18 out of the 22 isolates showed proteolytic activity on solid media with diameters of halo zone that ranged from 5 ± 0.71 mm for isolate coded as PSS-03 to 25 ± 1.41 mm for isolate coded as PSS-06. Intact chicken feathers were degraded by proteolytic bacterial isolates in variable degree that ranged from 24percent for PSS-10 and 81percent for DSS-02. Extracellular keratinase production recorded for the isolates ranged from 63.63 ± 4.14 U/mL for PSS-10 to 693.63 ± 62.99 U/mL for DSS-02. Based on 16S rDNA sequence and phylogenetic analysis, the 2 isolates with remarkable keratinolytic activity coded as DSS-02 and PSS-14 were identified as Chryseobacterium proteolyticum FGNn and Pseudomonas aeruginosa GNFx. C. proteolyticum showed the maximum keratinase production of 756.36 U/mL after 72 h of incubation at optimized fermentation conditions which involved initial medium pH (4), incubation temperature (30 oC), inoculum size (2percent; v/v), and chicken feathers (1.5percent; w/v). Similarly, P. aeruginosa optimally produced keratinase (1055.45 U/mL) after 96 h of incubation at optimized fermentation conditions that involved initial medium pH (7-8), incubation temperature (30 oC), inoculum size (5percent; v/v), and chicken feathers (2.5percent; w/v). Furthermore, feather hydrolysate from C. proteolyticum FGNn had relatively higher abundance (>1.5g/100g sample) of arginine (1.85), serine (1.63), glycine (1.9) and lysine (1.62); while P. aeruginosa GNFx feather hydrolysate showed high abundance of arginine, serine, aspartic acid, glutamic acid, glycine, alanine, valine, and leucine with respective concentration of 2.06, 1.67, 2.39, 3.05, 1.87, 1.73, 1.56 and 1.65 (g/100g sample). The results showed that keratinases from the two bacterial isolates were optimally active at pH 8, and temperature of 50 oC for FGNn keratinase and 50-60 oC for GNFx keratinase. The enzymes displayed remarkable pH stability. Keratinase from C. proteolyticum was catalytically inhibited by EDTA and 1,10-phenanthroline but not affected by PMSF; while P. aeruginosa keratinase was not significantly affected by those class of protease inhibitors. Adiitionally, FGNn keratinase demonstrated high residual activity of 90percent, 103percent, 101percent, 110percent, 130, and 105percent in the presence of DTT, hydrogen peroxides, acetonitrile, triton X-100, tween-80 and SDS, respectively. Similarly, catalytic efficiency of GNFx keratinase was promoted in the presence of hydrogen peroxides (119percent), triton X-100 (140percent), tween-80 (150percent) and SDS (147percent) compared to the control. Furthermore, the keratinases from the both bacterial isolates exhibited catalytic efficiency enhancement and remarkable structural stability in the presence of laundry detergents tested. The findings from the study suggest the application potentials of the isolates for the bioconversion of recalcitrant keratinous wastes into digestible and quality protein hydrolysates. The properties of these microbial keratinases indicate that they may be exploited for various biotechnological and industrial processes especially in the formulation of detergents.Thesis (MSc) -- Faculty of Science and Agriculture, 202

Manipulating DNA and RNA structures via click-to-release caged nucleic acids for biological and biomedical applications

Effectively controlling the structures of DNA and RNA is crucial for their functional utilization in material development, biological regulation, and medical applications. Here, we present a gain-of-function strategy for controlling DNA and RNA structures using an inverse electron-demand Diels-Alder (IEDDA) based click-to-release reaction. By incorporating click reaction-cleavable caged moiety into oligonucleotides, we disrupt activated base pairs, allowing controlled release of biofunctional higher-order nucleic acid structures. This click-to-release caged DNA was employed to control DNA duplex formation. Next, we demonstrated the utility of "click-to-release"strategy for regulated release of Z-DNA or Z-RNA and bind associated proteins. In addition, the approach was used to manipulated G-quadruplex formation in vitro and in vivo, enabling visual detection of G-quadruplex using BVE-caged DNA with fluorescent dye. Furthermore, we demonstrated the utility of click-to-release caged DNA for Quantum Dots (QDs) functionalization, enabling precise molecular imaging for cancer diagnosis. Finally, we developed a click-to-release controllable nucleic acid aptamer for precise blood clotting regulation and anticoagulation therapy. This strategy provides moderate kinetics, excellent orthogonality, and biocompatibility. It establishes a new pathway towards control of nucleic acid structures and functions, which has promising applications in various biological procedures and nucleic acid medicines

Histological differences among thrombi in thrombotic diseases

Citation: Atsushi Yamashita, Toshihiro Gi, Yuichiro Sato, Histological differences among thrombi in thrombotic diseases, Current Opinion in Hematology, 32(3), 146-156, 2025-02-07, https://doi.org/10.1097/MOH.000000000000086

Continuous Exposure of Nonobese Adult Male Rats to a Soft-Textured, Readily Absorbable Diet Induces Insulin Resistance and Derangements in Hepatic Glucose and Lipid Metabolism

Background Type 2 diabetes (T2D) is characterized by insulin resistance and defective insulin secretion. Previously, we found that rats fed soft pellets (SPs) on a 3-h restricted schedule over 14 wk demonstrated glucose intolerance and insulin resistance with disruption of insulin signaling. Objectives This study aims to determine 1) the time required for an SP diet to induce insulin resistance, and 2) whether the metabolic derangements in rats fed SPs can be reversed by changing to a standard control diet. Methods We performed glucose tolerance tests and calculated the homeostasis model assessment of insulin resistance (HOMA-IR) to evaluate the insulinemic response to glucose and assess insulin resistance in nonobese male rats fed control pellets (CPs) or SPs on a 3–h restricted schedule (10:00–13:00) for 4 and 9 wk. At 11 wk, we switched half of the insulin-resistant SP group to CPs [soft-to-control pellets (SCPs)] and after an additional 11 wk evaluated changes in glucose and lipid metabolism across the 3 groups. Results The glucose tolerance test results in the SP and CP rats did not differ at 4 or 9 wk. The insulin levels in the SP group were higher than in the CP group at both time points (P < 0.05). The HOMA-IR was significantly higher in the SP rats at 9 wk compared with the controls (P < 0.05). At 22 wk, the HOMA-IR, blood glucose levels at 30 min after initiating feeding, hepatic glucose metabolism, and lipid synthesis in rats fed SPs continuously were significantly greater than in those fed CPs (P < 0.05); however, these values in the SCP rats did not differ from those in the CP rats. Conclusions A continuous diet of soft-textured, readily absorbable food may be an important and reversible underlying driver in T2D pathogenesis

広島県三次市下本谷遺跡最高所地点の発掘調査 -後期旧石器時代前半期台形様石器群の検討-

Articledepartmental bulletin pape

Ecto-nucleoside triphosphate diphosphohydrolase inhibits ATP- and ADP-induced vasoconstriction

http://www.elsevier.com/wps/find/journaldescription.cws_home/369/descriptio

Application of fiber tractography and diffusion tensor imaging to evaluate spinal cord diseases in dogs

Fiber tractography is a technique capable of depicting the three-dimensional structure and connectivity of nerve fibers using serial magnetic resonance diffusion tensor imaging (DTI). To establish fiber tractography and DTI methods in veterinary clinical medicine, we evaluated fiber tractography and DTI parameters: apparent diffusion coefficient (ADC) values and fractional anisotropy (FA) values, in various spinal cord diseases. Spinal cord DTI was examined in 28 dogs with spinal cord diseases. The ADC and FA values were measured at lesion sites and cranial normal sites on spinal cords, and both values of lesion sites were compared with normal sites. In thoracolumbar intervertebral disk herniation (IVDH) cases, depending on their neurologic grades, fiber tractography indicated rupture of fiber trajectories, loss of neuronal bundles and disorder of fiber directions. In these cases, the average ADC values at lesion sites significantly decreased compared with normal sites (P=0.016). In the progressive myelomalacia case, the average ADC and FA values of hyperintense swollen regions in T2WI decreased compared to both values in other disease cases. Finally, in the meningioma case, the continuity of fiber trajectories improved after the administration of an anticancer agent. This study suggests that fiber tractography and DTI are useful in the diagnosis and prognosis of veterinary spinal cord diseases.Citation: Konishi Y, Satoh H, Kuroiwa Y, Kusaka M, Yamashita A, Asada Y, Asanuma T. Application of fiber tractography and diffusion tensor imaging to evaluate spinal cord diseases in dogs. J Vet Med Sci. 2017 Feb 28;79(2):418-424. doi: 10.1292/jvms.16-0504. Epub 2016 Dec 23. PMID: 28025450; PMCID: PMC5326951

Altered choline level in atherosclerotic lesions: Upregulation of choline transporter-like protein 1 in human coronary unstable plaque

Inflammatory activity and hypoxia in atherosclerotic plaques are associated with plaque instability and thrombotic complications. Recent studies show that vascular cell metabolism affects atherogenesis and thrombogenicity. This study aimed to identify the metabolites in macrophage-rich unstable plaques that modulate atherogenesis and serve as potential markers of plaque instability. Atherosclerotic plaques were induced by balloon injury in the iliofemoral arteries of rabbits fed on a conventional or 0.5% cholesterol diet. At 3 months post-balloon injury, the arteries and cardiac tissues were subjected to histological, quantitative real-time polymerase chain reaction, and metabolomic analyses. The identified metabolite-related proteins were immunohistochemically analyzed in stable and unstable plaques from human coronary arteries. The factors modulating the identified metabolites were examined in macrophages derived from human peripheral blood mononuclear cells. Metabolomic analysis revealed that choline and guanine levels in macrophage-rich arteries were upregulated compared with those in non-injured arteries and cardiac tissues. Vascular choline levels, but not guanine levels, were positively correlated with the areas immunopositive for macrophages and tumor necrosis factor (TNF)-α and matrix metalloproteinase (MMP) 9 mRNA levels in injured arteries. In human coronary arteries, choline transporter-like protein (CTL) 1 was mainly localized to macrophages within plaques. The area that was immunopositive for CTL1 in unstable plaques was significantly higher than that in stable plaques. Intracellular choline levels were upregulated upon stimulation with TNF-α but were downregulated under hypoxia in cultured macrophages. Administration of choline upregulated the expression of TNF-α and CTL1 mRNA in cultured macrophages. The transfection of CTL1 small interfering RNA decreased CTL1, TNF-α, and MMP9 mRNA levels in cultured macrophages. These results suggest that choline metabolism is altered in macrophage-rich atherosclerotic lesions and unstable plaques. Thus, CTL1 may be potential markers of plaque instability.Citation: Nakamura E, Maekawa K, Saito Y, Matsumoto T, Ogawa M, Komohara Y, Asada Y, Yamashita A. Altered choline level in atherosclerotic lesions: Upregulation of choline transporter-like protein 1 in human coronary unstable plaque. PLoS One. 2023 Feb 17;18(2):e0281730. doi: 10.1371/journal.pone.0281730. PMID: 36800352; PMCID: PMC9937458