Korea Research Institute of Bioscience and Biotechnology
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Evaluation of anti-inflammatory effect by regulating NF-κB pathway of Argyreia capitata (Vahl) Choisy extract in LPS-induced RAW 264.7 macrophages
No full textArgyreia capitata (Vahl) Choisy is a species of the genus Argyreia. Although many studies have analyzed the biological activity of A. nervosa, A. speciasa, and A. acuta, the anti-inflammatory effect of A. capitata extract (ACE) remains unclear. In this study, we evaluated the anti-inflammatory effect of ACE using lipopolysaccharide (LPS)-induced inflammatory markers in RAW 264.7 cells. We confirmed that the ACE inhibited the LPS-induced NO (nitric oxide) and iNOS (inducible nitric oxide synthase) expression in RAW 264.7 cells. ACE suppressed not only the LPS-induced phosphorylation of IKK, IκB, and p65 but also IL-1β expression. Collectively, these results suggest that ACE is a novel anti-inflammatory agent that suppresses iNOS expression, NO production, and the NF-κB signaling pathways.
Validation of marker-assisted selection in soybean breeding program for pod shattering resistance
No full textSignificant yield losses are caused by pod shattering during the harvest of domesticated crop plants. The phenotype evaluation of pod shattering requires fully grown plants, and it is a labor-intensive and time-consuming procedure. The purpose of this study was to test the applicability of previously identified KSS-SNP5 SNP marker on chromosome 16 in the marker assisted selection (MAS) system for pod shattering in soybean breeding programs. For this research, TaqMan SNP assay was successfully developed to detect the A/G allele of the KSS-SNP5 marker in the breeding materials, including two breeding stages, F2:3 populations, and yield trial stage breeding lines. The MAS prediction accuracy was calculated by comparing the genotyping data from the TaqMan assay with the phenotype data from the dry-oven method. The total MAS prediction accuracy was 92.5% and 96.2% in two F2:3 populations and breeding lines, respectively and 85% and 96% accuracy were shown in severe conditions. In this study, we confirmed that the KSS-SNP5 TaqMan probe tightly linked to pod shattering is effective to select a resistant line. Based on these results, we propose that the marker tightly linked to pod shattering can be of valuable use in soybean breeding program for the development of pod shattering-resistant cultivars.
CD9 induces cellular senescence and aggravates atherosclerotic plaque formation
No full textCD9, a 24？kDa tetraspanin membrane protein, is known to regulate cell adhesion and migration, cancer progression and metastasis, immune and allergic responses, and viral infection. CD9 is upregulated in senescent endothelial cells, neointima hyperplasia, and atherosclerotic plaques. However, its role in cellular senescence and atherosclerosis remains undefined. We investigated the potential mechanism for CD9-mediated cellular senescence and its role in atherosclerotic plaque formation. CD9 knockdown in senescent human umbilical vein endothelial cells significantly rescued senescence phenotypes, while CD9 upregulation in young cells accelerated senescence. CD9 regulated cellular senescence through a phosphatidylinositide 3 kinase-AKT-mTOR-p53 signal pathway. CD9 expression increased in arterial tissues from humans and rats with age, and in atherosclerotic plaques in humans and mice. Anti-mouse CD9 antibody noticeably prevented the formation of atherosclerotic lesions in ApoE？/？ mice and Ldlr？/？ mice. Furthermore, CD9 ablation in ApoE？/？ mice decreased atherosclerotic lesions in aorta and aortic sinus. These results suggest that CD9 plays critical roles in endothelial cell senescence and consequently the pathogenesis of atherosclerosis, implying that CD9 is a novel target for prevention and treatment of vascular aging and atherosclerosis.
Variation in some characters of the genus Sceliphron (Hymenoptera: Sphecidae) in the Philippines
No full textThe observed morphological and color variations for Sceliphron species in the Philippines are hereby reported. Sceliphron madraspatanum conspicillatum and Sceliphron rufopictum laticinctum have several variations in propodeal maculation pattern and in pronotal collar and petiole of the former. Postmortem color changes were also observed in S. r. laticinctum. A single yellow band on the third tergite of S. r. laticinctum is distinct when it is alive, and an additional yellow band develops on the posterior part of abdominal tergites 2-5 after death. Mandibles may either be sharp or blunt, and clypeus may be lobed or flat for S. r. laticinctum and Sceliphron javanum aemulum. However, newly emerged adults of these species have sharper mandibles and lobed clypeus. DNA barcoding was used to address intraspecific color and morphological variation of the Philippine Sceliphrini, but not all specimens were successfully barcoded because most of the specimens were very old and contaminated by fungi. However, this study came up with a new primer that has successfully barcoded 3% of the specimens.
Detection and assessment of α-synuclein oligomers in the urine of Parkinson's disease patients
No full textBackground: α-Synuclein (α-syn) is a major component of Lewy bodies, a pathologic marker of Parkinson's disease (PD) in post-mortem studies. The use of α-syn as a practical PD biomarker has been investigated by numerous researchers. However, reports of differences in α-syn levels in biofluids, such as cerebrospinal fluid, plasma, and saliva, between PD patients and controls are inconsistent. Recently, the measurement of α-syn oligomer levels has emerged as a novel approach to diagnose PD.
Objective: Lysates and culture media from two different types of dopaminergic neuronal cells or urine samples from 11 non-PD and 21 PD patients were collected and analyzed.
Methods: We developed and performed an enzyme-linked immuno-absorbent assay (ELISA) to detect various oligomeric α-syn using distinct pairs of antibodies.
Results: We validated our ELISA using rotenone-induced alterations of α-syn levels in human dopaminergic neurons. Total urinary α-syn levels, measured using our ELISA method, showed no difference between PD and non-PD individuals, but a higher level of α-syn oligomer recognized by MJFR-14-6-5-2 in PD urine samples was observed. Levels of distinct oligomeric α-syn detected by ASyO5 were lower in PD urine samples. Three different α-syn ELISA results were analyzed with respect to the severity of PD, but only the correlation between total α-syn levels and PD index was significant.
Conclusion: Our findings suggest that detection of distinct oligomeric formations of α-syn and measurement of their levels in urine might be feasible for use in PD diagnostics.
Clinical characteristics of ataxia-telangiectasia presenting dystonia as a main manifestation
No full textIntroduction: Besides cerebellar ataxia, various other movement disorders, including dystonia, could manifest as main clinical symptoms in ataxia-telangiectasia (A-T). However, the clinical characteristics of dystonic A-T patients are not clearly elucidated.
Methods: To investigate the characteristics of dystonic A-T, we screened previous reports with A-T patients presenting dystonia as a main manifestation, and included 38 dystonic A-T patients from 16 previous studies and our 2 cases. We reviewed clinical and demographic data of dystonic A-T patients. Additionally, to figure out clinical meaning of cerebellar involvement in dystonic A-T, we divided them into two groups based on the presence of cerebellar involvement, and compared clinical features between two groups.
Results: In the patients with dystonic A-T, dystonia tended to appear during childhood or adolescence and became generalized over time. Choreoathetosis and myoclonus accompanied more frequently than the typical clinical features, including cerebellar ataxia or atrophy, telangiectasia, or oculomotor apraxia. Additionally, alpha-fetoprotein level was also elevated in the patients with dystonic A-T. When we compared dystonic A-T with and without cerebellar involvement, the former was related with more chance for telangiectasia and oculomotor apraxia, while the latter with that for choreoathetosis and malignancy.
Conclusion: Even without ataxia, telangiectasia, or oculomotor apraxia, A-T should be considered in undiagnosed dystonia, especially generalized dystonia which started from childhood or adolescence period, and alpha-fetoprotein level can be a useful screening tool. In addition, cerebellar involvement is important considering different phenotype in dystonic A-T patients with and without cerebellar sign.
Effects of a titanium dioxide thin film for improving the biocompatibility of diamond-like coated coronary stents
No full textThis study was performed to evaluate the effects of a diamond-like carbon (DLC) layer on the biological properties of a pre-treated titanium dioxide (TiO2) coronary stent surface. TiO2 and DLC were coated onto a cobalt-chromium stent surface in consecutive order using the plasma-enhanced chemical vapor deposition method (TiO2/DLC). To investigate the effect of TiO2, a group treated only with DLC was also prepared as a control. The surface characteristics were investigated by SEM, ESCA, AFM, and contact angle analyses. To estimate the effect of surface modification on biological response, endothelial cell migration was carried out. As a result, cracking of the coating layer on the strut curved portion in the DLC group was restored in the TiO2/DLC group. ESCA result showed that the peaks sp2 and sp3 were slightly shifted and higher in the TiO2/DLC group. The surface roughness average was highly increased in the TiO2/DLC group (Ra; 18.033) compared to the BMS (Ra; 9.258) and DLC groups (Ra; 10.144). By the increment of Ra, the surface property became hydrophilic in the TiO2/DLC group (39.8°？±？5.88°) compared to the non-treated (72.7°？±？2.24°) and DLC (78.6°？±？2.24°) groups. Endothelial cell migration was significantly increased in the TiO2/DLC group (68.9？±？13.52%) compared to the non-treated (5.4？±？3.14%) and DLC groups (22.3？±？8.22%). Therefore, the improvement of surface properties and endothelial cell migration in the TiO2/DLC group could improve the physiological response in the human body.
Inhibitory effect of α-ketoglutaric acid on α-glucosidase: integrating molecular dynamics simulation and inhibition kinetics
No full textThe inhibition of α-glucosidase is used as a key clinical approach to treat type 2 diabetes mellitus and thus, we assessed the inhibitory effect of α-ketoglutaric acid (AKG) on α-glucosidase with both an enzyme kinetic assay and computational simulations. AKG bound to the active site and interacted with several key residues, including ASP68, PHE157, PHE177, PHE311, ARG312, TYR313, ASN412, ILE434 and ARG439, as detected by protein？ligand docking and molecular dynamics simulations. Subsequently, we confirmed the action of AKG on α-glucosidase as mixed-type inhibition with reversible and rapid binding. The relevant kinetic parameter IC50 was measured (IC50 = 1.738？±？0.041？mM), and the dissociation constant was determined (Ki Slope = 0.46？±？0.04？mM). Regarding the relationship between structure and activity, a high AKG concentration induced the slight modulation of the shape of the active site, as monitored by hydrophobic exposure. This tertiary conformational change was linked to AKG inhibition and mostly involved regional changes in the active site. Our study provides insight into the functional role of AKG due to its structural property of a hydroxyphenyl ring that interacts with the active site. We suggest that similar hydroxyphenyl ring-containing compounds targeting key residues in the active site might be potential α-glucosidase inhibitors.
Recent insights into Aeromonas salmonicida and its bacteriophages in aquaculture: a comprehensive review
No full textThe emergence and spread of antimicrobial resistance in pathogenic bacteria of fish and shellfish have caused serious concerns in the aquaculture industry, owing to the potential health risks to humans and animals. Among these bacteria, Aeromonas salmonicida, which is one of the most important primary pathogens in salmonids, is responsible for significant economic losses in the global aquaculture industry, especially in salmonid farming because of its severe infectivity and acquisition of antimicrobial resistance. Therefore, interest in the use of alternative approaches to prevent and control A. salmonicida infections has increased in recent years, and several applications of bacteriophages (phages) have provided promising results. For several decades, A. salmonicida and phages infecting this fish pathogen have been thoroughly investigated in various research areas including aquaculture. The general overview of phage usage to control bacterial diseases in aquaculture, including the general advantages of this strategy, has been clearly described in previous reviews. Therefore, this review specifically focuses on providing insights into the phages infecting A. salmonicida, from basic research to biotechnological application in aquaculture, as well as recent advances in the study of A. salmonicida.
Loss of HSPA9 induces peroxisomal degradation by increasing pexophagy
No full textQuality control of peroxisomes is essential for cellular homeostasis. However, the mechanism underlying pexophagy is largely unknown. In this study, we identified HSPA9 as a novel pexophagy regulator. Downregulation of HSPA9 increased macroautophagy/autophagy but decreased the number of peroxisomes in vitro and in vivo. The loss of peroxisomes by HSPA9 depletion was attenuated in SQSTM1-deficient cells. In HSPA9-deficient cells, the level of peroxisomal reactive oxygen species (ROS) increased, while inhibition of ROS blocked pexophagy in HeLa and SH-SY5Y cells. Importantly, reconstitution of HSPA9 mutants found in Parkinson disease failed to rescue the loss of peroxisomes, whereas reconstitution with wild type inhibited pexophagy in HSPA9-depleted cells. Knockdown of Hsc70-5 decreased peroxisomes in Drosophila, and the HSPA9 mutants failed to rescue the loss of peroxisomes in Hsc70-5-depleted flies. Taken together, our findings suggest that the loss of HSPA9 enhances peroxisomal degradation by pexophagy.