Korea Research Institute of Bioscience and Biotechnology
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Biological and computational studies for dual cholinesterases inhibitory effect of zerumbone
No full textAcetylcholinesterase (AChE) and butyrylcholinesterase (BChE) mediate the degradation of acetylcholine (ACh), a primary neurotransmitter in the brain. Cholinergic deficiency occurs during the progression of Alzheimer’s disease (AD), resulting in widespread cognitive dysfunction and decline. We evaluated the potential effect of a natural cholinesterase inhibitor, zerumbone, using in vitro target enzyme assays, as well as in silico docking and ADMET (absorption, distribution, metabolism, excretion, and toxicity) simulation. Zerumbone showed a predominant cholinesterase inhibitory property with IC50 values of 2.74 ± 0.48 μM and 4.12 ± 0.42 μM for AChE and BChE, respectively; however, the modes of inhibition were different. Computational docking simulation indicated that Van der Waals interactions between zerumbone and both the cholinesterases were the main forces responsible for its inhibitory effects. Furthermore, zerumbone showed the best physicochemical properties for both bioavailability and blood？brain barrier (BBB) permeability. Together, in the present study, zerumbone was clearly identified as a unique dual AChE and BChE inhibitor with high permeability across the BBB, suggesting a strong potential for its physiological benefits and/or pharmacological efficacy in the prevention of AD.
Infection and rapid transmission of SARS-CoV-2 in ferrets
No full textThe outbreak of coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in China and rapidly spread worldwide. To prevent SARS-CoV-2 dissemination, understanding the in vivo characteristics of SARS-CoV-2 is a high priority. We report a ferret model of SARS-CoV-2 infection and transmission that recapitulates aspects of human disease. SARS-CoV-2-infected ferrets exhibit elevated body temperatures and virus replication. Although fatalities were not observed, SARS-CoV-2-infected ferrets shed virus in nasal washes, saliva, urine, and feces up to 8 days post-infection. At 2 days post-contact, SARS-CoV-2 was detected in all naive direct contact ferrets. Furthermore, a few naive indirect contact ferrets were positive for viral RNA, suggesting airborne transmission. Viral antigens were detected in nasal turbinate, trachea, lungs, and intestine with acute bronchiolitis present in infected lungs. Thus, ferrets represent an infection and transmission animal model of COVID-19 that may facilitate development of SARS-CoV-2 therapeutics and vaccines. ⓒ 2020 Elsevier Inc.The outbreak of coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) rapidly spreads, leading to a pandemic infection. Kim et al. show that ferrets are highly susceptible to SARS-CoV-2 infection and effectively transmit the virus by direct or indirect contact, recapitulating human infection and transmission.
Iroquois homeobox protein 2 identified as a potential Biomarker for Parkinson’s disease
No full textThe diagnosis of Parkinson’s disease (PD) is initiated after the occurrence of motor symptoms, such as resting tremors, rigidity, and bradykinesia. According to previous reports, non-motor symptoms, notably gastrointestinal dysfunction, could potentially be early biomarkers in PD patients as such symptoms occur earlier than motor symptoms. However, connecting PD to the intestine is methodologically challenging. Thus, we generated in vitro human intestinal organoids from PD patients and ex vivo mouse small intestinal organoids from aged transgenic mice. Both intestinal organoids (IOs) contained the human LRRK2 G2019S mutation, which is the most frequent genetic cause of familial and sporadic PD. By conducting comprehensive genomic comparisons with these two types of IOs, we determined that a particular gene, namely, Iroquois homeobox protein 2 (IRX2), showed PD-related expression patterns not only in human pluripotent stem cell (PSC)-derived neuroectodermal spheres but also in human PSC-derived neuronal cells containing dopaminergic neurons. We expected that our approach of using various cell types presented a novel technical method for studying the effects of multi-organs in PD pathophysiology as well as for the development of diagnostic markers for PD.
Butylparaben is toxic to porcine oocyte maturation and subsequent embryonic development following in vitro fertilization
No full textParabens are widely used in personal care products due to their antimicrobial effects. Although the toxicity of parabens has been reported, little information is available on the toxicity of butylparaben (BP) on oocyte maturation. Therefore, we investigated the effects of various concentrations of BP (0 μM, 100 μM, 200 μM, 300 μM, 400 μM, and 500 μM) on the in vitro maturation of porcine oocytes. BP supplementation at a concentration greater than 300 μM significantly reduced the proportion of complete cumulus cell expansion and metaphase II oocytes compared to the control. The 300 μM BP significantly decreased fertilization, cleavage, and blastocyst formation rates with lower total cell numbers and a higher rate of apoptosis in blastocysts compared to the control. The BP-treated oocytes showed significantly higher reactive oxygen species (ROS) levels, and lower glutathione (GSH) levels than the control. BP significantly increased the aberrant mitochondrial distribution and decreased mitochondrial function compared to the control. BP-treated oocytes exhibited significantly higher percentage of γ-H2AX, annexin V-positive oocytes and expression of LC3 than the control. In conclusion, we demonstrated that BP impaired oocyte maturation and subsequent embryonic development, by inducing ROS generation and reducing GSH levels. Furthermore, BP disrupted mitochondrial function and triggered DNA damage, early apoptosis, and autophagy in oocytes.
Recent updates on outbreaks of shiga toxin-producing Escherichia coli and its potential reservoirs
No full textFollowing infection with certain strains of Shiga toxin-producing Escherichia coli (STEC), particularly enterohemorrhagic ones, patients are at elevated risk for developing life-threatening extraintestinal complications, such as acute renal failure. Hence, these bacteria represent a public health concern in both developed and developing countries. Shiga toxins (Stxs) expressed by STEC are highly cytotoxic class II ribosome-inactivating proteins and primary virulence factors responsible for major clinical signs of Stx-mediated pathogenesis, including bloody diarrhea, hemolytic uremic syndrome (HUS), and neurological complications. Ruminant animals are thought to serve as critical environmental reservoirs of Stx-producing Escherichia coli (STEC), but other emerging or arising reservoirs of the toxin-producing bacteria have been overlooked. In particular, a number of new animal species from wildlife and aquaculture industries have recently been identified as unexpected reservoir or spillover hosts of STEC. Here, we summarize recent findings about reservoirs of STEC and review outbreaks of these bacteria both within and outside the United States. A better understanding of environmental transmission to humans will facilitate the development of novel strategies for preventing zoonotic STEC infection.
Ochrobactrum soli sp. nov., isolated from a Korean cattle farm
No full textA Gram stain negative, motile, non-spore-forming, rod-shaped, strictly aerobic, beige-pigmented bacterium, designated strain BO-7T, was isolated from soil of cattle farm, in Seosan, Republic of Korea. On the basis of 16S rRNA gene sequencing, strain BO-7T clustered with species of the genus Ochrobactrum and appeared closely related to O. haematophilum CCUG 38531T (98.9%), O. daejeonense KCTC 22458T (98.1%), O. rhizosphaerae DSM 19824T (98.1%), O. pituitosum DSM 22207T (98.0%), and O. pecoris DSM 23868T (98.0%). The digital DNA-DNA hybridization and average nucleotide identity between strain BO-7T and the closely related strains were 21.9-39.1%, 78.5-89.5%, respectively, indicating that BO-7T is a novel species of the genus Ochrobactrum. The DNA G + C content of the genomic DNA was 57.1 mol%, and ubiquinone Q-10 was the predominant respiratory quinone. The polar lipids consisted of phosphatidylglycerol, phosphatidylethanolamine, phosphatidylcholine, phosphatidylmonomethyl-ethanolamine, di-phosphatidylglycerol, the major polyamines were spermidine, putrescine, and sym-homospermidine. The major cellular fatty acids (> 5%) were C16:0, C19:0 cycle ω7c, and C18:1ω7c and/or C18:1ω6c (summed feature 8). ANI calculation, digital DNA-DNA hybridization, physiological and biochemical characteristics indicated that strain BO-7T represents a novel species of the genus Ochrobactrum, for which the name Ochrobactrum soli sp. nov. is proposed. The type strain is BO-7T (= KACC 19676T = LMG 30809T).
Potential cause of decrease in bloom events of the harmful dinoflagellate Cochlodinium polykrikoides in Southern Korean coastal waters in 2016
No full textBlooms of the ichthyotoxic dinoflagellate Cochlodinium polykrikoides are responsible for massive fish mortality events in Korean coastal waters (KCW). They have been consistently present in southern KCW over the last two decades, but they were not observed in 2016, unlike in the previous years. Despite extensive studies, the cause of this absence of this dinoflagellate bloom remains largely unknown. Thus, we compared physico-chemical and biological data from along the Tongyeong coast between 2016 and the previous four years (2012？2015). The averages of water temperature and salinity in August, 2016 were significantly (p < 0.001) different from those in the previous years. The amount of Changjiang River discharge, which can affect the environmental conditions in the southern Korean coastal area via ocean currents, was larger than in the previous years, resulting in a reduction in the salinity level in August when blooms of C. polykrikoides usually occurred. Moreover, compared to previous years, in 2016, there was a weak expansion of C. polykrikoides blooms in the Goheung-Oenarodo area where C. polykrikoides blooms were annually initiated in KCW. Lastly, the strong winds from the typhoon Lionrock may also have contributed to the early termination of this dinoflagellate bloom. Together with these findings, the combination of these environmental conditions in 2016, unlike in previous years, may have inhibited the formation of C. polykrikoides blooms along the Tongyeong coast.
Genomic and histopathological characteristics of Vibrio parahaemolyticus isolated from an acute hepatopancreatic necrosis disease outbreak in Pacific white shrimp (Penaeus vannamei) cultured in Korea
No full textThis report describes the detailed microbial and histological characteristics of Vibrio parahaemolyticus strains associated with acute hepatopancreatic necrosis disease (VpAHPND) that infected Pacific white shrimp (Penaeus vannamei) cultured in Korea. Shrimps showing typical symptoms of AHPND, including a whitish hepatopancreas and empty gut, were reported at two private shrimp farms in Korea in 2019. In the hepatopancreas of the field samples, sloughing of undifferentiated cells in the hepatopancreatic tubule epithelium was observed upon histopathological examination, and 2 Vibrio spp.-related toxin genes (pirA and pirB) associated with AHPND were detected in the samples. Among samples obtained from the hepatopancreas, pond water, and feces (pond sediment), the copy number of the virulent AHPND plasmid was the highest in the hepatopancreas samples, followed by that in the pond water and feces (pond sediment) samples. Two VpAHPND strains were isolated from pond water and diseased shrimp. The genome of the virulent AHPND plasmid pVp_Kor-D1？2 (GenBank accession no. CP046414.1), isolated from strain 19？021-D1, was fully sequenced. We confirmed that the Korean VpAHPND strain harbored the Asian-type AHPND plasmid, which was almost identical to the AHPND-associated V. owensii plasmid pVOWZ2 previously identified in China. Furthermore, the pathogenicity of the Korean VpAHPND strains was evaluated in laboratory bioassays, and we verified that those isolates were highly pathogenic to Pacific white shrimp, causing high cumulative mortality within 3 days.
Reclassification of Gonyaulax verior (Gonyaulacales, Dinophyceae) as Sourniaea diacantha gen. et comb. nov.
No full textGonyaulax verior was initially described as Amylax diacantha from Belgian coastal waters a century ago but its detailed morphology needed restudy. Here, we established nine strains of G. verior by germinating cysts or isolating cells from localities from the European Atlantic to the Caspian Sea and the Pacific Ocean. Both cyst and thecal morphology were examined by light and scanning electron microscopy. SSU, LSU and/or ITS-5.8S rRNA gene sequences were obtained from all strains. Cells of G. verior have a plate formula of Po, 4′, 2a, 6-7′′, 6C, 6S, 6′′′, 1p, 1′′′′ with an L-type ventral organisation, characterised by two either straight or curved antapical horns of variable length. Cysts of G. verior are oval, smooth and contain one or two yellow accumulation bodies. The maximum-likelihood and Bayesian inference analyses based on SSU and LSU rRNA gene sequences revealed two clades of G. verior, referred to as ribotypes A and B. Genetic distances based on ITS-5.8S rRNA gene sequences within the same ribotype were less than 0.06, but greater than 0.32 between ribotypes. G. verior is reclassified as Sourniaea diacantha gen. et comb. nov., which is attributed to Lingulodiniaceae together with Pyxidinopsis, Lingulodinium and Amylax. Our results suggest that Lingulodiniaceae can be separated from Protoceratiaceae and Gonyaulacaceae based on ventral organisation, apical complex, ventral pore and number of anterior intercalary plates. One strain of S. diacantha was examined for yessotoxin production by LC-MS/MS but did not produce toxin.
Picrasma quassioides extract elevates the cervical cancer cell apoptosis through ROS-mitochondrial axis activated p38 MAPK signaling pathway
No full textBackground/aim: Picrasma quassioides (P. quassioides) is used in traditional Asian medicine widely for the treatment of anemopyretic cold, eczema, nausea, loss of appetite, diabetes mellitus, hypertension etc. In this study we aimed to understand the effect of P. quassioides ethanol extract on SiHa cervical cancer cell apoptosis.
Materials and methods: The P. quassioides extract-induced apoptosis was analyzed using the MTT assay, fluorescence microscopy, flow cytometry and western blotting.
Results: P. quassioides extract induced cellular apoptosis by increasing the accumulation of cellular and mitochondrial reactive oxygen species (ROS) levels and inhibiting ATP synthesis. Pretreatment with N-Acetylcysteine (NAC), a classic antioxidant, decreased the intracellular ROS production and inhibited apoptosis. In addition, the P38 MAPK signaling pathway is a key in the apoptosis of SiHa cells induced by the P. quassioides extract.
Conclusion: The P. quassioides extract exerts its anti-cancer properties on SiHa cells through ROS-mitochondria axis and P38 MAPK signaling. Our data provide a new insight for P. quassioides as a therapeutic strategy for cervical cancer treatment.