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Phylosymbiosis is defined as the relationship in which the microbiome recapitulates the phylogeny of the host and has been demonstrated in a variety of terrestrial organisms, although it has been understudied in fish, the most phylogenetically diverse vertebrate. Given that the species-specificity of fish microbiomes was detected in multiple body parts and differed by body parts, we assumed that the phylogenetic reflection of the microbiome would differ across body parts. Thus, we analyze the difference of phylosymbiotic relationships in the microbial communities found in three body parts (skin, gills, and intestine) of seven wild fish species from four families (Labridae, Sebastidae, Sparidae, and Rajidae) via 16S rRNA gene amplicon sequencing. Fishes were purchased at Docheon port market in Tongyeong City, Korea and were transported to nearby research institutes for aliveness. Mantel tests using dissimilarity values of microbiomes and hosts' divergence times showed that the differences in microbial communities in all three body parts were related to the hosts' divergence time. This pattern was the most pronounced in the skin. Furthermore, fishes from the same family showed similar bacterial compositions on their skins and gills, with clear differences depending on the family, with the exception of Labridae. These results suggest that the skin microbiome is particularly vulnerable to evolutionary pressures. We hypothesized that the evolution of the fish immune system and the difference in feeding habits induced the stronger phylosymbiotic signal in the skin. Collectively, this dataset will be useful for understanding the fish microbiome and give insights into phylosymbiosis of aquatic animals across body parts., Competing Interests: Declarations. Ethics approval: All the experimental procedures were performed in accordance with the Institutional Animal Care and Use Committee (IACUC) guidelines for the care and use of animals. Han completed the course, ‘Usage and Managements of laboratory animals’ hosted by Bioethics Information Center (BIC). Yu, Kang, and Noh completed the course, ‘Usage and Managements of laboratory animals’ hosted by Korea Association For Laboratory Animals (KAFLA). Consent to Publish: All the authors consented to this paper’s publication. Competing Interests: The authors declare no competing interests., (© 2024. The Author(s).)

Previous studies showed no improvement in bacterial biomass for Candidatus Puniceispirillum marinum IMCC1322 under light regimes. Nevertheless, in nutrient-replete cultures with higher inoculating cell densities, strain IMCC1322 exhibited proteorhodopsin photoheterotrophy. Increasing both inoculum size and the amino acid pool can eliminate quorum sensing and starvation responses in strain IMCC1322. Light regimes affected IMCC1322 cultures in stationary/death phases, where cellular ATP levels ranged from 0.0331 to 1.74 mM, with ATP/cell ranging from 13.9 to 367 zeptomoles. In nutrient-depleted conditions, strain IMCC1322 may suffer from excessive protons generated by proteorhodopsin under light conditions. IMCC1322 may tolerate excessive periplasmic protons through ATP-dependent proton pumping and protonation of augmented amino acids. Meanwhile, acid stress could also be mitigated by refining membrane permeability through unsaturation and cyclopropanation of phospholipids. Oceanic bacteria such as IMCC1322 and SAR11 preferred anaplerotic TCA cycles over glycolysis and rely on the Entner-Doudoroff (ED) pathway for growth. Although ATP generation is less efficient in the ED pathway, it offers advantages during rapid growth owing to its strong thermodynamic driving force. The metabolism of IMCC1322 favors gluconeogenesis over glycolysis, aligning with the metabolism of SAR11 reported in previous studies. However, the additional light-driven, PR-dependent ATP synthesis in IMCC1322 is expected to be insufficient to support protein turnover after the log phase, as well as in nutrient-limited conditions. Stable isotope measurements showed no significant differences in the inorganic carbon assimilation between constant light and constant dark cultures in late log phase.

Background: To date, most intradialytic hypotension (IDH) studies have proposed technologies to comprehensively predict the occurrence of IDH using the patient's baseline information and ultrafiltration (UF) information, but it is difficult to apply the technologies while identifying the patient's condition in real time., Methods: In this study, we propose an IDH indicator that uses heart rate (HR) change information to identify the patient's condition in real time and visually shows whether IDH has occurred. The data used were collected from 40 dialysis patients in a clinical trial conducted in the Artificial Kidney Unit at Yeungnam University Medical Center, Korea, from 18 July to 29 November 2023., Results: The IDH indicator infers changes in the patient's blood pressure during dialysis by analyzing the upper and lower maximum HRs based on the real-time average HR. Medical staff can respond to IDH in real time by looking at the IDH indicator, which visually expresses changes in the patient's HR. In addition, we propose a multilayer perceptron structure that inputs upper and lower maximum HR information based on the average HR for the time interval accumulated in real time. In learning using 40 min of data up to 5 min before IDH occurred, models using two and five layers showed excellent performance, with accuracy of 88.6% and 85.2%, respectively., Conclusions: By combining IDH visual indicators and the multi-layer perceptron method, medical staff can effectively respond to IDH in real time.

Purpose: Breast cancer survivors following disease-modifying treatment frequently experience multiple-concurrent symptoms (Jansana et al. in Int J Cancer 149(10):1755 1767, 2021), negatively impacting their quality of life and increasing the risk of polypharmacy (Alwhaibi et al. in J Oncol Pharm Pract 26(5):1052 1059, 2020). This study evaluates the feasibility and acceptability of acupuncture for the management of the pain-fatigue-sleep disturbance-numbness/tingling symptom cluster in breast cancer survivors, and investigates relationships between the symptom cluster and Traditional Chinese Medicine (TCM) syndrome diagnosis., Methods: This was a single-arm, pre-test/post-test feasibility trial conducted at Chris O'Brien Lifehouse Hospital, Australia. Breast cancer survivors who completed treatment and experienced clinically significant levels of two or more symptoms (pain, fatigue, sleep disturbance, numbness/tingling) were eligible to participate in the individualized, pragmatic 6-week acupuncture intervention. The primary outcome was feasibility and acceptability. Effectiveness was explored using a symptom cluster mean score., Results: Twenty women enrolled in the study over an 11-week period and 90% completed the study. Most women agreed or completely agreed that acupuncture was feasible (85%), acceptable (90%), and appropriate (90%). Both mean and composite symptom cluster scores were significantly reduced (p < 0.001), as were individual symptom scores in fatigue (p < 0.001), sleep disturbance (p = 0.04), and numbness/tingling (p = 0.01). TCM syndromes most closely associated with this symptom cluster were Spleen qi deficiency and Heart fire. No adverse events were reported., Conclusion: This study demonstrated that acupuncture was safe and feasible, justifying a powered randomized control trial. Preliminary findings suggest beneficial effects of acupuncture for the management of the pain-fatigue-sleep disturbance-numbness/tingling symptom cluster for women with breast cancer. TCM syndromes identified in this trial may be used to guide acupuncture treatment protocols., Clinical Trial Registration: This trial was registered with the Australian New Zealand Clinical Trials Registry (ACTRN12622000590763) on 21 April 2022., (© 2024. The Author(s).)

The genus Bacteroides , a predominant group in the human gut microbiome, presents significant potential for microbiome engineering and the development of live biotherapeutics aimed at treating gut diseases. Despite its promising capabilities, tools for effectively engineering Bacteroides species have been limited. In our study, we have made a breakthrough by identifying novel signal peptides in Bacteroides thetaiotaomicron and Akkermansia muciniphila . These peptides facilitate efficient protein transport across cellular membranes in Bacteroides , a critical step for therapeutic applications. Additionally, we have developed an advanced episomal plasmid system. This system demonstrates superior protein secretion capabilities compared to traditional chromosomal integration plasmids, making it a vital tool for enhancing the delivery of therapeutic proteins in Bacteroides species. Initially, the stability of this episomal plasmid posed a challenge; however, we have overcome this by incorporating an essential gene-based selection system. This novel strategy not only ensures plasmid stability but also aligns with the growing need for antibiotic-free selection methods in clinical settings. Our work, therefore, not only provides a more robust secretion system for Bacteroides but also sets a new standard for the development of live biotherapeutics.

Brine shrimp (Artemia spp.) is a significant factor in determining aquaculture production. Since the microbiota of Artemia can colonize the gut in larvae, various microorganisms transmitted from Artemia can affect host larval health. Although the microbiota composition of Artemia would be essential in determining aquaculture productivity, our understanding on microbiome of Artemia is still insufficient. Through our study, we identified the species of Artemia cysts supplied by three different manufacturers (P1, P2, and P3) with investigation of size and hatching efficiency. The species of Artemia from P1 was identified as A. tibetiana, and P2 and P3 was A. franciscana. A. tibetiana hatched from the P1 cysts had the largest body size with the lowest hatching rate. Furthermore, we conducted a comprehensive analysis of the microbiome present in the rearing water and the nauplius whole body from each product. We observed specific microbiota compositions, both beneficial and harmful, depending on the product types and the sample types. Additionally, we found that the microbiota composition in the rearing water was associated with the manufacturing environment, while the compositions in the nauplius whole body were species-specific. Notably, we discovered that an extract containing microbiota from the nauplius sample of P3 increased the hatching rate of A. tibetiana, indicating a positive role in Artemia culture. These findings demonstrate that the microbial communities present in Artemia vary according to the product and/or species, underscoring their significance in aquaculture production., (© 2023. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

Isoprene has numerous industrial applications, including rubber polymer and potential biofuel. Microbial methane-based isoprene production could be a cost-effective and environmentally benign process, owing to a reduced carbon footprint and economical utilization of methane. In this study, Methylococcus capsulatus Bath was engineered to produce isoprene from methane by introducing the exogenous mevalonate (MVA) pathway. Overexpression of MVA pathway enzymes and isoprene synthase from Populus trichocarpa under the control of a phenol-inducible promoter substantially improved isoprene production. M. capsulatus Bath was further engineered using a CRISPR-base editor to disrupt the expression of soluble methane monooxygenase (sMMO), which oxidizes isoprene to cause toxicity. Additionally, optimization of the metabolic flux in the MVA pathway and culture conditions increased isoprene production to 228.1 mg/L, the highest known titer for methanotroph-based isoprene production. The developed methanotroph could facilitate the efficient conversion of methane to isoprene, resulting in the sustainable production of value-added chemicals., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Two novel Gram-stain-negative, strictly-aerobic, rod-shaped (1.2 ± 3.4 μm × 0.3 ± 0.7 μm), and non-motile marine bacterial species, designated MEBiC05379 T and MEBiC07777 T , were isolated from a marine sponge Pseudaxinella sp. in Gangneung City and deep-sea sediments of the Ulleung basin in the East Sea of Korea, respectively. The 16S rRNA gene sequence analysis revealed high levels of similarities between these strains and members of the genus Flavivirga (97.0-98.4% sequence identities). Both novel strains revealed as mesophilic, neutrophilic in pH and slightly halophilic. Similar to those of other Flavivirga members, the primary cellular fatty acids of both strains were iso-C 15:0 , iso-C 15:1 G, iso-C 15:03 -OH, and iso-C 17:0 3-OH, with MEBiC05379 T and MEBiC07777 T containing relatively higher proportions of C 12:0 and summed feature 3 (C 16:1 ω7c and/or C 16:1 ω6c). In both taxa, the major isoprenoid quinone was MK-6. The DNA G + C contents of MEBiC05379 T and MEBiC07777 T genomes were 32.62 and 32.46 mol%, respectively. Compared to other members of Flavivirga, both strains exhibited similar DNA G + C ratio and fatty acids pattern, yet enzyme expression and carbon sources utilization pattern were different. Genomes of the genus Flavivirga showed enzyme preferences to fucoidan and sulfated galactans. Considering the monophyly rule, AAI values delineate the genus Flavivirga from adjacent genera calculated to be 76.0-78.7%. Based on the phenotypic, genomic and biochemical data, strains for MEBiC05379 T and MEBiC07777 T thus represent two novel species in the genus Flavivirga, for which the names Flavivirga spongiicola sp. nov. (MEBiC05379 T [= KCTC 92527  T  = JCM 16662  T ]), and Flavivirga abyssicola sp. nov. (MEBiC07777 T [= KCTC 92563  T  = JCM 36477  T ]) are proposed., (© 2024. The Author(s), under exclusive licence to Microbiological Society of Korea.)

Background: Current endometriosis treatments do not always provide symptom relief, with many using complementary approaches. This study examined the effectiveness of acupuncture on pain and quality of life in people with endometriosis., Methods: Searches were conducted on Cochrane Central Register of Controlled Trials (CENTRAL), PubMed, Allied and Complementary Medicine Database (AMED) and Embase (Ovid), Epistemonikos, and Cumulative Index to Nursing and Allied Health Literature (CINAHL; EBSCOhost) on 20 March 2023. Trials were included if they used penetrating acupuncture. Risk of bias was assessed with Cochrane RoB2 and GRADE for overall evidence certainty. Random-effects meta-analyses were undertaken, using Hedges' g or mean difference (MD) both with 95 % confidence intervals (CI)., Results: Six studies involving a total of 331 participants were included. Evidence for benefit was found for acupuncture compared to non-specific acupuncture on overall pelvic pain ( g  = 1.54, 95 % CI 0.92 to 2.16, 3 RCTs, n  = 231, low certainty evidence, p <0.001), menstrual pain ( g  = 1.67, 95 % CI 1.23 to 2.12, 1 RCT, n  = 106, moderate certainty evidence, p <0.001), and non-specified pelvic pain (MD -2.77, 95 % CI 2.15 to 3.38, 2 RCTs, n  = 125, low certainty evidence, p <0.001), and compared to usual care on menstrual pain ( g  = 0.9, 95 % CI 0.15 to 1.64, 1 RCT, n  = 19, very low certainty evidence, p  = 0.02). Most studies reported low rates of adverse events., Conclusion: Acupuncture treatment for endometriosis demonstrated clinically relevant improvements in pelvic pain and should be considered as a potential treatment intervention., Study Registration: PROSPERO ID: CRD42023408700., (© 2023 Korea Institute of Oriental Medicine. Published by Elsevier B.V.)

Long Fiber Spray-up Molding (LFSM) deviates from the conventional approach in liquid composite molding (LCM) processes by utilizing extremely long chopped strands of fibers as the primary reinforcement material in its fabrication process. In LFSM, chopped fibers are impregnated with resin that is sprayed vertically downwards before reaching the mold surface. The spraying mechanism is mounted on an actuator, which is capable of spraying freely in any specified pattern or direction. Under LFSM, it is extremely difficult to fabricate a composite part with uniformly distributed fiber content throughout its volume. The consequences of the non-uniform fiber volume distribution arise from the fiber entanglement as the length of the fiber reaches up to 100 mm in LFSM. In this study, the effect of fiber entanglement during LFSM was analyzed through various approaches. This included measuring the coefficient of friction between fibers in contact and examining the correlation between fiber lengths and the number of intersections. Furthermore, the viscoelastic properties of the uncured composite part were assessed by experimenting with the influence of viscosity on fiber length during compression molding. The results were then computed, modeled, and visualized in MATLAB, considering variations in viscosity and fiber length, both before and after compression molding., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2023 Published by Elsevier Ltd.)

Acetogenic bacteria can utilize C1 compounds, such as carbon monoxide (CO), formate, and methanol, via the Wood-Ljungdahl pathway (WLP) to produce biofuels and biochemicals. Two novel acetogenic bacteria of the family Eubacteriaceae ES2 and ES3 were isolated from Eulsukdo, a delta island in South Korea. We conducted whole genome sequencing of the ES strains and comparative genome analysis on the core clusters of WLP with Acetobacterium woodii DSM1030 T and Eubacterium limosum ATCC8486 T . The methyl-branch cluster included a formate transporter and duplicates or triplicates copies of the fhs gene, which encodes formyl-tetrahydrofolate synthetase. The formate dehydrogenase cluster did not include the hydrogenase gene, which might be replaced by a functional complex with a separate electron bifurcating hydrogenase (HytABCDE). Additionally, duplicated copies of the acsB gene, encoding acetyl-CoA synthase, are located within or close to the carbonyl-branch cluster. The serum bottle culture showed that ES strains can utilize a diverse range of C1 compounds, including CO, formate, and methanol, as well as CO 2 . Notably, ES2 exhibited remarkable resistance to high concentrations of C1 substrates, such as 100% CO (200 kPa), 700 mM formate, and 500 mM methanol. Moreover, ES2 demonstrated remarkable growth rates under 50% CO (0.45 h -1 ) and 200 mM formate (0.34 h -1 ). These growth rates are comparable to or surpassing those previously reported in other acetogenic bacteria. Our study introduces novel acetogenic ES strains and describes their genetic and physiological characteristics, which can be utilized in C1-based biomanufacturing., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Yu, Park, Lee, Kwon, Lim, Lee, Kang, Lee, Kwon and Kim.)

It is important to improve cerebrovascular health before the occurrence of cerebrovascular disease, as it has various aftereffects and a high recurrence rate, even with appropriate treatment. Various medical recommendations for preventing cerebrovascular diseases have been introduced, including smoking cessation, exercise, and diet. However, the effectiveness of these methods varies greatly from person to person, and their effects cannot be confirmed unless they are practiced over a long period. Therefore, there is a growing need to develop more quantitative methods that are applicable to the public to promote cerebrovascular health. Thus, in this study, we aimed to develop noninvasive and quantitative thermal stimulation techniques using ultrasound to improve cerebrovascular health and prevent cerebrovascular diseases. This study included 27 healthy adults in their 20s (14 males, 13 females). Thermal stimulation using therapeutic ultrasound at a frequency of 3 MHz was applied to the right sternocleidomastoid muscle in the supine posture for 2 min at four intensities (2.4, 5.1, 7.2, and 10.2 W/cm 2 ). Diagnostic ultrasound was used to measure the peak systolic velocity (PSV), heart rate (HR), and pulse wave velocity (PWV) in the right common carotid artery (CCA), and the physiological changes were compared between intervention intensities. Compared to pre-intervention (preI), the PSV showed a significant increase during intervention (durI) at intensities of 7.2 W/cm 2 and 10.2 W/cm 2 ( p = 0.010 and p = 0.021, respectively). Additionally, PWV showed a significant decrease for post-intervention (postI) at 7.2 W/cm 2 and 10.2 W/cm 2 ( p = 0.036 and p = 0.035, respectively). However, the HR showed no significant differences at any of the intensities. The results demonstrate that an intervention at 3 MHz with an intensity of 7.2 W/cm 2 or more can substantially increase cerebral blood flow and reduce arterial stiffness. Therefore, the use of therapeutic ultrasound of appropriate intensity is expected to improve the cerebral blood flow and reduce vascular stiffness to maintain cerebral blood flow at a certain level, which is closely related to the prevention and treatment of cerebrovascular diseases, thereby improving cerebrovascular health.

Levulinic acid (LA) is a valuable chemical used in fuel additives, fragrances, and polymers. In this study, we proposed possible biosynthetic pathways for LA production from lignin and poly(ethylene terephthalate). We also created a genetically encoded biosensor responsive to LA, which can be used for screening and evolving the LA biosynthesis pathway genes, by employing an LvaR transcriptional regulator of Pseudomonas putida KT2440 to express a fluorescent reporter gene. The LvaR regulator senses LA as a cognate ligand. The LA biosensor was first examined in an Escherichia coli strain and was found to be non-functional. When the host of the LA biosensor was switched from E. coli to P. putida KT2440, the LA biosensor showed a linear correlation between fluorescence intensity and LA concentration in the range of 0.156-10 mM LA. In addition, we determined that 0.156 mM LA was the limit of LA detection in P. putida KT2440 harboring an LA-responsive biosensor. The maximal fluorescence increase was 12.3-fold in the presence of 10 mM LA compared to that in the absence of LA. The individual cell responses to LA concentrations reflected the population-averaged responses, which enabled high-throughput screening of enzymes and metabolic pathways involved in LA biosynthesis and sustainable production of LA in engineered microbes.

A strictly anaerobic hyperthermophilic archaeon, designated strain IOH2 T , was isolated from a deep-sea hydrothermal vent (Onnuri vent field) area on the Central Indian Ocean Ridge. Strain IOH2 T showed high 16S rRNA gene sequence similarity to Thermococcus sibiricus MM 739 T (99.42 %), Thermococcus alcaliphilus DSM 10322 T (99.28 %), Thermococcus aegaeus P5 T (99.21 %), Thermococcus litoralis DSM 5473 T (99.13 %), ' Thermococcus bergensis' T7324 T (99.13 %), Thermococcus aggregans TY T (98.92 %) and Thermococcus prieurii Bio-pl-0405IT2 T (98.01 %), with all other strains showing lower than 98 % similarity. The average nucleotide identity and in silico DNA-DNA hybridization values were highest between strain IOH2 T and T. sibiricus MM 739 T (79.33 and 15.00 %, respectively); these values are much lower than the species delineation cut-offs. Cells of strain IOH2 T were coccoid, 1.0-1.2 µm in diameter and had no flagella. Growth ranges were 60-85 °C (optimum at 80 °C), pH 4.5-8.5 (optimum at pH 6.3) and 2.0-6.0 % (optimum at 4.0 %) NaCl. Growth of strain IOH2 T was enhanced by starch, glucose, maltodextrin and pyruvate as a carbon source, and elemental sulphur as an electron acceptor. Through genome analysis of strain IOH2 T , arginine biosynthesis related genes were predicted, and growth of strain IOH2 T without arginine was confirmed. The genome of strain IOH2 T was assembled as a circular chromosome of 1 946 249 bp and predicted 2096 genes. The DNA G+C content was 39.44 mol%. Based on the results of physiological and phylogenetic analyses, Thermococcus argininiproducens sp. nov. is proposed with type strain IOH2 T (=MCCC 4K00089 T =KCTC 25190 T ).

The engineered Methylococcus capsulatus Bath presents a promising approach for converting methane, a potent greenhouse gas, into valuable chemicals. High cell-density culture (HCDC) is necessary for high-titer growth-associated bioproducts, but it often requires time-consuming and labor-intensive optimization processes. In this study, we aimed to achieve efficient HCDC of M. capsulatus Bath by measuring the residual nutrient levels during bioreactor operations and analyzing the specific uptake of each medium component. By controlling the concentrations of nutrients, particularly calcium and phosphorus via intermittent feeding, we achieved a high cell density of 28.2 g DCW/L and a significantly elevated production of mevalonate at a concentration of 1.8 g/L from methane. Our findings demonstrate that the methanotroph HCDC approach presented herein offers a promising strategy for promoting sustainable development, with an exceptional g-scale production titer for value-added synthetic biochemicals.

Two Gram-stain-positive, motile, endospore-forming, facultatively anaerobic strains, designated MEBiC13591 T and MEBiC13594 T , were isolated from tidal flat sediment of the Incheon City on the west coast of Korea. Growth of both novel strains was observed at pH 5-9 (optimum, pH 7-7.5), and in 0-8% NaCl (optimum, 2% for MEBiC13591 T and 3% for MEBiC13594 T ). Strains MEBiC13591 T and MEBiC13594 T grew optimally at 50 °C, (37.5-56.1 °C) and 44 °C (20.7-50.7 °C), respectively. The main cellular fatty acids of strain MEBiC13591 T were iso-C 15: 0 , anteiso-C 15: 0 , iso-C 16: 0 , iso-C 17: 0 and anteiso-C 17: 0 , while those for strain MEBiC13594 T were C 14: 0 , iso-C 14: 0 , iso-C 15: 0 , anteiso-C 15: 0 and C 16: 0 . In both taxa, the major isoprenoid was MK-7. The genomic DNA G + C contents were 34.1 and 37.0 mol% for MEBiC13591 T and MEBiC13594 T , respectively. A 16S rRNA gene sequence analysis revealed that the novel strains showed high similarity with members of the genera Aeribacillus (95.0%) and Caldibacillus (93.5-94.5%); however, showed lower than 90% with Caldibacillus debilis Tf T . Phylogenetic and Phylogenomic analysis revealed that two novel strains comprised distinct phyletic line with members formerly assigned to Caldibacillus. Based on genomic indices, such as AAI and ANI, members formerly affiliated with Caldibacillus and Bacillus as well as the novel strains should be classified into five independent genera. Based on the phenotypic, genomic and biochemical data, strains MEBiC13591 T and MEBiC13594 T represent two novel species in the novel genus, for which the names Fervidibacillus albus gen. nov., sp. nov. (MEBiC13591 T [= KCCM 43317 T  = KCTC 43181 T  = JCM 33662 T  = MCCC 1K04565 T ]), and Fervidibacillus halotolerans sp. nov. (MEBiC13594 T [= KCTC 43182 T  = JCM 34001 T ]) are proposed. Three additional genera Caldifermentibacillus, Palidibacillus, and Perspicuibacillus are also proposed by reclassification of the several species with valid names that formerly affiliated with the genera Caldibacillus., (© 2023. The Author(s), under exclusive licence to Microbiological Society of Korea.)

Antibiotics have been widely used for plasmid-mediated cell engineering. However, continued use of antibiotics increases the metabolic burden, horizontal gene transfer risks, and biomanufacturing costs. There are limited approaches to maintaining multiple plasmids without antibiotics. Herein, we developed an inverter cascade using CRISPRi by building a plasmid containing a single guide RNA (sgRNA) landing pad (pSLiP); this inhibited host cell growth by repressing an essential cellular gene. Anti-sgRNAs on separate plasmids restored cell growth by blocking the expression of growth-inhibitory sgRNAs in pSLiP. We maintained three plasmids in Escherichia coli with a single antibiotic selective marker. To completely avoid antibiotic use and maintain the CRISPRi-based logic inverter cascade, we created a novel d-glutamate auxotrophic E. coli. This enabled the stable maintenance of the plasmid without antibiotics, enhanced the production of the terpenoid, (-)-α-bisabolol, and generation of an antibiotic-resistance gene-free plasmid. CRISPRi is therefore widely applicable in genetic circuits and may allow for antibiotic-free biomanufacturing., (© The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research.)

This study aimed to characterise and evaluate the probiotic properties of a newly isolated marine bacterium, strain S6031. The isolated strain was identified as Pseudoalteromonas ruthenica. In vivo experiments were conducted with P. ruthenica-immersed larvae and P. ruthenica-enriched Artemia fed to adult zebrafish. Disease tolerance of larval zebrafish against Edwardsiella piscicida was demonstrated by 66.34% cumulative per cent survival (CPS) in the P. ruthenica-exposed group, which was higher than the CPS of the control (46.67%) at 72 h post challenge (hpc). Heat-stressed larvae had 55% CPS in the P. ruthenica-immersed group, while the control had 30% CPS at 60 hpc. Immune-stress response gene transcripts (muc5.1, muc5.2, muc5.3, alpi2, alpi3, hsp70, and hsp90a) were induced, while pro-inflammatory genes (tnfα, il1b, and il6) were downregulated in P. ruthenica-immersed larvae compared to the control. This trend was confirmed by low pro-inflammatory and high stress-responsive protein expression levels in P. ruthenica-exposed larvae. Adult zebrafish had higher CPS (27.2%) in the P. ruthenica-fed group than the control (9.52%) upon E. piscicida challenge, suggesting increased disease tolerance. Histological analysis demonstrated modulation of goblet cell density and average villus height in the P. ruthenica-supplemented group. Metagenomics analysis clearly indicated modulation of alpha diversity indices and the relative abundance of Proteobacteria in the P. ruthenica-supplemented zebrafish gut. Furthermore, increased Firmicutes colonisation and reduced Bacteroidetes abundance in the gut were observed upon P. ruthenica supplementation. Additionally, this study confirmed the concentration-dependent increase of colony dispersion and macrophage uptake upon mucin treatment. In summary, P. ruthenica possesses remarkable functional properties as a probiotic that enhances host defence against diseases and thermal stress., Competing Interests: Declaration of competing interest The authors declare that they have no conflicts of interest., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

We sequenced the complete mitochondrial genome of sand dollar Astriclypeus mannii (Verrill 1867) (Echinoidea: Astriclypeidae) occurring in the subtidal sand flat in Jeju Island off the south coast of Korea. The mitochondrial genome was 15,744 bp in length and contained 13 protein-coding genes (PCGs), 22 transfer RNAs, two ribosomal RNAs, and 140 nucleotides representing the putative control region. We reconstructed the concatenated phylogenetic tree based on 13 PCGs of 18 echinoderms, including A. mannii . From the maximum likelihood clustering, A. mannii was grouped in the order Echinolampadacea. The complete mitochondrial sequence of A. mannii for the first time in this study provide valuable insight in understanding the evolution and phylogenetic analysis of echinoids (sea urchins)., Competing Interests: No potential competing interest was reported by the author(s)., (© 2022 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

This study presents a novel DNA part characterization technique that increases throughput by combinatorial DNA part assembly, solid plate-based quantitative fluorescence assay for phenotyping, and barcode tagging-based long-read sequencing for genotyping. We confirmed that the fluorescence intensities of colonies on plates were comparable to fluorescence at the single-cell level from a high-end, flow-cytometry device and developed a high-throughput image analysis pipeline. The barcode tagging-based long-read sequencing technique enabled rapid identification of all DNA parts and their combinations with a single sequencing experiment. Using our techniques, forty-four DNA parts (21 promoters and 23 RBSs) were successfully characterized in 72 h without any automated equipment. We anticipate that this high-throughput and easy-to-use part characterization technique will contribute to increasing part diversity and be useful for building genetic circuits and metabolic pathways in synthetic biology.

Intradialytic hypotension (IDH) is a common side effect that occurs during hemodialysis and poses a great risk for dialysis patients. Many studies have been conducted so far to predict IDH, but most of these could not be applied in real-time because they used only underlying patient information or static patient disease information. In this study, we propose a multilayer perceptron (MP)-based IDH prediction model using heart rate (HR) information corresponding to time-series information and static data of patients. This study aimed to validate whether HR differences and HR slope information affect real-time IDH prediction in patients undergoing hemodialysis. Clinical data were collected from 80 hemodialysis patients from 9 September to 17 October 2020, in the artificial kidney room at Yeungnam University Medical Center (YUMC), Daegu, South Korea. The patients typically underwent hemodialysis 12 times during this period, 1 to 2 h per session. Therefore, the HR difference and HR slope information within up to 1 h before IDH occurrence were used as time-series input data for the MP model. Among the MP models using the number and data length of different hidden layers, the model using 60 min of data before the occurrence of two layers and IDH showed maximum performance, with an accuracy of 81.5%, a true positive rate of 73.8%, and positive predictive value of 87.3%. This study aimed to predict IDH in real-time by continuously supplying HR information to MP models along with static data such as age, diabetes, hypertension, and ultrafiltration. The current MP model was implemented using relatively limited parameters; however, its performance may be further improved by adding additional parameters in the future, further enabling real-time IDH prediction to play a supporting role for medical staff.

A novel anaerobic, rod-shaped, non-motile bacterium, designated strain ES005 T , was isolated from tidal flat sediments near the rhizosphere of Phragmites australis at Eulsukdo Island, Republic of Korea. A polyphasic approach revealed that cells of the strain were Gram-stain-positive, catalase- and oxidase-negative, non-spore-forming rods. Phylogenetic analyses based on 16S rRNA gene sequences revealed that strain ES005 T belonged to the family Eubacteriaceae , class Clostridia and showed the highest sequence similarity to Alkalibacter mobili s (97.52 %) and followed by Alkalibacter saccharofermentans Z-79820 T (96.72%). The OrthoANI value between strain ES005 T and A. mobilis was 69.67 %. Strain ES005 T grew optimally at 33-37 °C, at pH 6.0-7.0 and in the presence of 1-2 % (w/v) NaCl. Growth in 12.5 % CO atmosphere was observed. Acetate and formate were end products of fructose fermentation and growth on CO. The major cellular fatty acids of strain ES005 T were C 14 : 0 (39.1 %) and C 16 : 0 (26.6 %). The major polar lipids were diphoshatidylgycerol, phosphatidylglycerol and three unidentified phospholipids. The DNA G+C content of strain ES005 T was 46.9 mol%. Based on the phenotypic, phylogenetic, genomic and chemotaxonomic features of the isloate, strain ES005 T represents a novel species, for which the name Alkalibacter rhizosphaerae sp. nov. is proposed. The type strain is ES005 T (=KCTC 25246 T =JCM 34530 T ).

Bacteria in the Desulfovibrionaceae family, which contribute to S element turnover as sulfate-reducing bacteria (SRB) and disproportionation of partially oxidized sulfoxy anions, have been extensively investigated since the importance of the sulfur cycle emerged. Novel species belonging to this taxon are frequently reported, because they exist in various environments and are easy to culture using established methods. Due to the rapid expansion of the taxon, correction and reclassification have been conducted. The development of high-throughput sequencing facilitated rapid expansion of genome sequence database. Genome-based criteria, based on these databases, proved to be potential classification standard by overcoming the limitations of 16S rRNA-based phylogeny. Although standards methods for taxogenomics are being established, the addition of a novel genus requires extensive calculations with taxa, including many species, such as Desulfovibrionaceae. Thus, the genome-based criteria for classification of Desulfovibrionaceae were established and validated in this study. The average amino-acid identity (AAI) cut-off value, 63.43 ± 0.01, was calculated to be an appropriate criterion for genus delineation of the family Desulfovibrionaceae. By applying the AAI cut-off value, 88 genomes of the Desulfovibrionaceae were divided into 27 genera, which follows the core gene phylogeny results. In this process, two novel genera ( Alkalidesulfovibrio and Salidesulfovibrio ) and one former invalid genus (" Psychrodesulfovibrio ") were officially proposed. Further, by applying the 95-96% average nucleotide identity (ANI) standard and the 70% digital DNA-DNA hybridization standard values for species delineation of strains that were classified as the same species, five strains have the potential to be newly classified. After verifying that the classification was appropriately performed through relative synonymous codon usage analysis, common characteristics were listed by group. In addition, by detecting metal resistance related genes via in silico analysis, it was confirmed that most strains display metal tolerance., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Park, Kim, Park, Yu, Namirimu, Roh and Kwon.)

A novel bacterium, designated SCR006 T , was isolated from tidal flat sediment from Suncheon Bay, Republic of Korea. Cells of strain SCR006 T were strictly anaerobic, motile cocci, Gram-reaction-negative, and catalase- and oxidase-negative. Growth was observed at 4-41 °C (optimum, 34-37 °C), at pH 6.5-10.0 (optimum, pH 7.0-7.5) and in presence of 0-8 % NaCl (optimum, 0-2 %). Fermentation products of peptone-yeast-glucose medium were acetate and ethanol. Results of phylogenetic analyses based on 16S rRNA gene sequences indicated that strain SCR006 T had high sequence similarity to Proteiniclasticum ruminis D3RC-2 T (97.9 %), followed by Youngiibacter multivorans DSM 6139 T (95.9 %) and Youngiibacter fragilis 232.1 T (95.0 %). The average nucleotide identity value between strain SCR006 T and P. ruminis DSM 24773 T was 72.7 %, which strongly supported that strain SCR006 T reresents a novel species within the genus Proteiniclasticum . The major cellular fatty acids are iso-C 15 : 0 (27.2 %) and anteiso-C 15 : 0 (16.9 %). The polar lipids were diphosphatidylglycerol, phosphatidylglycerol, two unidentified phospholipids, an unidentified aminolipid and five unidentified lipids. The genomic size was 3.2 Mb with genomic DNA G+C content of 45.6 mol%. The results of 16S rRNA-based and genome-based phylogenetic tree analyses indicated that SCR006 T should be assigned to the genus Proteiniclasticum . Strain SCR006 T could be distinguished from P. ruminis D3RC-2 T by its growth conditions, cell morphology and genomic characteristics. Based on the phenotypic, phylogenetic, genomic and chemotaxonomic features, strain SCR006 T represents a novel species, for which the name Proteiniclasticum aestuarii sp. nov. is proposed, with the type strain SCR006 T (=KCTC 25245 T = JCM 34531 T ).

Objectives: Efforts to safely reduce hospital LOS while maintaining quality outcomes and patient satisfaction are paramount. The primary goal of this study was to assess trends in LOS at a high-volume quaternary care spine center. Secondary goals were to assess trends in factors most associated with prolonged LOS., Methods: This is a prospective study of all consecutive patients admitted from January 2006 to December 2019. Data included demographics, diagnostic category (degenerative, oncology, deformity, trauma, other), LOS (mean, median, interquartile range, standard deviation, defined as days from admission to discharge), and in-hospital adverse events., Results: A total of 13,493 patients were included. Overall LOS has not changed over time with an overall median of 6.3 days (p = 0.451). Median LOS significantly increased for patients treated for degenerative pathology from 2.2 days in 2006 to 3.2 days in 2019 (p = 0.019). LOS has not changed for patients treated for deformity (overall median 6.8 days, p = 0.411), oncology (overall median 11.0 days, p = 0.051), or trauma (overall median 11.8 days, p = 0.582). Emergency admissions increased 3.2%/year for degenerative pathologies (p =  < 0.001). Mean age has increased from 48.4 years in 2006 to 58.1 years in 2019 (p =  < 0.001). This trend was observed in the deformity, degenerative and trauma group, not for patients treated for oncological disease. More adverse events were significantly associated with increasing age., Conclusion: This is the first North American study to comprehensively analyze trends in LOS for spinal surgery overtime in an academic center. Overall, LOS has not changed from 2006-2019. Various factors that influence LOS appear to have balanced each other. It may also be explained by the changing epidemiology of both elective and emergency surgeries. These findings provide opportunities for intervention and improvement, targeted at the geriatric population, to reduce length of hospitalization., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Enantiomerically pure d-amino acids are important intermediates as chiral building blocks for peptidomimetics and semisynthetic antibiotics. Here, a transcriptional factor-based screening strategy was used for the rapid screening of d-stereospecific amino acid amidase via an enzyme-specific amidophenol substrate. We used a d-threonine amidophenyl derivative to produce 2-aminophenol that serves as a putative enzyme indicator in the presence of d-threonine amidases. Comparative analyses of known bacterial species indicated that several Bacillus strains produce amidase and form putative indicators in culture media. The estimated amidase was cloned and subjected to rapid directed evolution through biosensor cells. Consequently, we characterized the F119A mutation that significantly improved the catalytic activity toward d-alanine, d-threonine, and d-glutamate. Its beneficial effects were confirmed by higher conversions and recurrent applications of the mutant enzyme, compared to the wild-type. This study showed that rapid directed evolution with biosensors coupled to designed substrates is useful to develop biocatalytic processes.

A Gram-stain-negative, aerobic, rod-shaped (0.3-0.5 × 1.0-1.9 µm), non-motile marine bacterium designated as ALE3EI T was isolated from a saline volcanic rock aquifer (lava sea-water) on Jeju Island, Republic of Korea. The 16S rRNA gene sequence analysis revealed that strain ALE3EI T showed high similarity to 'Altibacter lentus' JLT2010 T (97.2%), followed by Marixanthomonas ophiurae KMM 3046 T (94.5%). Growth was observed at 10-41°C (optimum, 30°C), at pH 6.0-8.5 (optimum, pH 7.5) and at 0.5-8% (optimum, 4.0%) NaCl. The predominant cellular fatty acids were iso-C 15:0 (23.5%), iso-C 16:0 (10.2%), iso-C 16:0 3OH (10.5%), and iso-C 17:0 3OH (16.8%). The DNA G + C contents was 40.4 mol%. The major respiratory quinone was MK-6. The major polar lipids were determined to be phosphatidylethanolamine, two unidentified glycolipids, and two unidentified aminolipids. Several phenotypic characteristics such as production of acetoin, activities of arginine dihydrolase and acid phosphatase, and utilization pattern of carbon sources differentiate strain ALE3EI T from 'A. lentus' JLT2010 T . Activities of the lipase, trypsin, α-chymotrypsin and gelatinase and utilization pattern of carbon sources differentiate strain ALE3EI T from M. ophiurae KMM 3046 T . The genome of strain ALE3EI T is 3.0 Mbp long and its ANI and AAI values against 'A. lentus' JLT2010 T were 76.58 and 72.76, respectively, however, AAI values against members in other genera were lower than 72%. The phylogenomic tree inferred by PhyloPhlAn clearly differentiated the strain ALE3EI T together with strain JLT2010 T from other genera in the Falvobacteriaceae. This polyphasic taxonomic data indicates that strain ALE3EI T should be identified as a novel species in the genus 'Altibacter', however, the name has not been validated. Therefore, the strain is classified as a novel genus and is proposed as Constantimarinum furrinae gen. nov., sp. nov. The type strain is ALE3EI T (= KCCM 43303 T = JCM 33022 T )., (© 2022. The Microbiological Society of Korea.)

Purpose: Around three quarters of individuals undergoing chemotherapy self-report multiple symptoms. There is clinical trial evidence of effectiveness for acupuncture for commonly experienced symptoms, and emerging evidence for reflexology, but little is known about the effects of these therapies on multiple symptoms when implemented in a real world setting during active chemotherapy treatment., Methods: This was a cohort study of participants receiving reflexology and/or acupuncture while attending chemotherapy. Participants received a 20 minute reflexology treatment or a 20 minute acupuncture treatment or a combination of both. Patient reported outcome measures were administered before and after the treatment using the Edmonton Symptom Assessment Scale (ESAS)., Results: During the study period, 330 unique patients received a total of 809 acupuncture and/or reflexology treatments. Participants had, on average, 5.3 symptoms each which they reported as moderate to severe (≥4/10) using the ESAS at baseline. Following treatment, participants reported 3.2 symptoms as moderate to severe. The symptom change for all participant encounters receiving any therapy was statistically significant for all symptoms, and clinically significant (a reduction of more than 1) for all symptoms except financial distress, appetite, and memory. Clinically significant levels of global distress (&lt;3) were reduced in 72% of all participants receiving either therapy. No adverse events were recorded., Conclusions: The results indicate that acupuncture and reflexology administered alongside chemotherapy may reduce patient reported symptom burden and patient global symptom related distress. Future research would include an active control group, and consider confounding factors such as chemotherapy stage and medication.

The characteristics or aspects of important fiducial points (FPs) in the electrocardiogram (ECG) signal are complicated because of various factors, such as non-stationary effects and low signal-to-noise ratio. Due to the various noises caused by the ECG signal measurement environment and by typical ECG signal deformation due to heart diseases, detecting such FPs becomes a challenging task. In this study, we introduce a novel PQRST complex detector using a one-dimensional bilateral filter (1DBF) and the temporal characteristics of FPs. The 1DBF with noise suppression and edge preservation preserves the P- or T-wave whereas it suppresses the QRS-interval. The 1DBF acts as a background predictor for predicting the background corresponding to the P- and T-waves and the remaining flat interval excluding the QRS-interval. The R-peak and QRS-interval are founded by the difference of the original ECG signal and the predicted background signal. Then, the Q- and S-points and the FPs related to the P- and T-wave are sequentially detected using the determined searching range and detection order based on the detected R-peak. The detection performance of the proposed method is analyzed through the MIT-BIH database (MIT-DB) and the QT database (QT-DB).

Methanol dehydrogenase (Mdh), is a crucial enzyme for utilizing methane and methanol as carbon and energy sources in methylotrophy and synthetic methylotrophy. Engineering of Mdh, especially NAD-dependent Mdh, has thus been actively investigated to enhance methanol conversion. However, its poor catalytic activity and low methanol affinity limit its wider application. In this study, we applied a transcriptional factor-based biosensor for the direct evolution of Mdh from Lysinibacillus xylanilyticus (Lxmdh), which has a relatively high turnover rate and low K M value compared to other wild-type NAD-dependent Mdhs. A random mutant library of Lxmdh was constructed in Escherichia coli and was screened using formaldehyde-detectable biosensors by incubation with low methanol concentrations. Positive clones showing higher fluorescence were selected by fluorescence-activated cell sorting (FACS) system, and their catalytic activities toward methanol were evaluated. The successfully isolated mutants E396V, K318N, and K46E showed high activity, particularly at very low methanol concentrations. In kinetic analysis, mutant E396V, K318N, and K46E had superior methanol conversion efficiency, with 79-, 23-, and 3-fold improvements compared to the wild-type, respectively. These mutant enzymes could thus be useful for engineering synthetic methylotrophy and for enhancing methanol conversion to various useful products.

Genetic circuits have been developed for quantitative measurement of enzyme activity, metabolic engineering of strain development, and dynamic regulation of microbial cells. A genetic circuit consists of several bio-elements, including enzymes and regulatory cassettes, that can generate the desired output signal, which is then used as a precise criterion for enzyme screening and engineering. Antagonists and inhibitors are small molecules with inhibitory effects on regulators and enzymes, respectively. In this study, an antagonist and an inhibitor were applied to a genetic circuit for a dynamic detection range. We developed a genetic circuit relying on regulators and enzymes, allowing for straightforward control of its output signal without additional genetic modification. We used para -nitrophenol and alanine as an antagonist of DmpR and inhibitor of tyrosine phenol-lyase, respectively. We show that the antagonist resets the detection range of the genetic circuit similarly to a resistor in an electrical logic circuit. These biological resistors in genetic circuits can be used as a rapid and precise controller of variable outputs with minimal circuit configuration., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Kwon, Kim, Yeom, Rha, Lee, Lee, Lee and Lee.)

A Gram-stain-negative, facultatively anaerobic, rod-shaped (1.8-4.4×0.5-0.7 µm) and motile marine bacterium, designated as MEBiC13590 T , was isolated from tidal flat sediment sampled at Incheon City, on the west coast of the Republic of Korea. The 16S rRNA gene sequence analysis revealed that strain MEBiC13590 T showed high similarity to Oricola cellulosilytica CC-AMH-0 T (98.2 %), followed by Oceaniradius stylonematis StC1 T (97.5 %); however, it clustered with Oricola cellulosilytica . The phylogenomic tree inferred by the up-to-date bacterial core gene set suggested that strain MEBiC13590 T shared a phyletic line with Oricola cellulosilytica . Average nucleotide identity and digital DNA-DNA hybridization values (75.0 and 19.3 %, respectively) between strain MEBiC13590 T and Oricola cellulosilytica CC-AMH-0 T were below the respective species delineation cutoffs. Growth was observed at 22-50 °C (optimum, 45 °C), at pH 5-9 (optimum, pH 7) and with 1-6 % (optimum, 3 %) NaCl. The predominant cellular fatty acids were C 16 : 0 (7.6 %), C 18 : 0 (12.2 %), 11-methyl C 18 : 1  ω7 c (5.7 %), C 19 : 0 cyclo ω6 c and summed feature 8 (comprising C 18 : 1  ω7 c and/or C 18 : 1  ω6 c ; 38 %). The DNA G+C content was 63.5 mol%. The major respiratory quinone was Q-10. Several phenotypic characteristics such as growth temperature, oxygen requirement, enzyme activities of urease, gelatinase, lipase (C 14 ), α-chymotrypsin, acid phosphatase, β-galactosidase, β-glucosidase etc. differentiate strain MEBiC13590 T from Oricola cellulosilytica CC-AMH-0 T . Based on this polyphasic taxonomic data, strain MEBiC13590 T should be classified as representing a novel species in the genus Oricola for which the name Oricola thermophila sp. nov. is proposed . The type strain is MEBiC13590 T (=KCCM 43313 T =JCM 33661 T ).

Bacteria initiate complicated signaling cascades from the detection of intracellular metabolites or exogenous substances by hundreds of transcription factors, which have been widely investigated as genetically-encoded biosensors for molecular recognition. However, the limited number of transcription factors and their broad substrate specificity result in ambiguity in small molecule identification. This study presents a new small molecule fingerprinting technique using evolutionary biosensor arrays with a machine learning technique that can capture highly specific substrate signals. Employing multiple mutant transcription factors derived from a single transcription factor has effectively circumvented the limited availability of transcription factors induced by a small molecule of our interest. This method achieved up to 95.3% true positive rate for identifying small molecules, and the high-resolution protein engineering technique improved the limit of detection 75-fold. The signal trade-offs with background noises caused by the complex cellular biochemistry of mutant transcription factors enable the biosensor arrays to be more informative in terms of statistical variance. The machine learning technology, coupled with the single transcription factor-driven evolutionary biosensor array, will open new avenues for molecular fingerprinting technologies., (Copyright © 2020 Elsevier B.V. All rights reserved.)

With the advancement of the Internet of Medical Things technology, many vital sign-sensing devices are being developed. Among the diverse healthcare devices, portable electrocardiogram (ECG) measuring devices are being developed most actively with the recent development of sensor technology. These ECG measuring devices use different sampling rates according to the hardware conditions, which is the first variable to consider in the development of ECG analysis technology. Herein, we propose an R-point detection method using an adaptive median filter based on the sampling rate and analyze major arrhythmias using the signal characteristics. First, the sliding window and median filter size are determined according to the set sampling rate, and a wider median filter is applied to the QRS section with high variance within the sliding window. Then, the R point is detected by subtracting the filtered signal from the original signal. Methods for detecting major arrhythmias using the detected R point are proposed. Different types of ECG signals were used for a simulation, including ECG signals from the MIT-BIH arrhythmia database and MIT-BIH atrial fibrillation database, signals generated by a simulator, and actual measured signals with different sampling rates. The experimental results indicated the effectiveness of the proposed R-point detection method and arrhythmia analysis technique.

Cardiac rehabilitation after percutaneous coronary intervention decreases recurrence and mortality but has a high dropout rate. The aim of this study is to identify dropout predictors by comparing the characteristics of complete and dropout patients in cardiac rehabilitation. The study included 593 patients (455 men and 138 women) who received percutaneous coronary intervention and were enrolled in a 1-year cardiac rehabilitation program consisting of home-based cardiac rehabilitation with three center visits. Dropout was defined as participation in the first center visit but not the second or third center visits. Blood lipids, quality of life, socioeconomic status, and 6-minute walk distance measurements at the first visit were compared between participants who completed and dropped out of cardiac rehabilitation. For both men and women, the dropout rate significantly correlated with a low 6-minute walk distance and low muscle mass ratio. The dropout rate was significantly higher for men, but not women, with low education and low income. However, the dropout rate was decreased for women, but not men, with low blood pressure and triglycerides. An improved understanding of the characteristics of participants and the cardiac rehabilitation dropout rate are expected to contribute to the development of cardiac rehabilitation strategies that decrease patient dropout.

Background: The novel coronavirus (coronavirus disease 2019 [COVID-19]) outbreak began in China in December last year, and confirmed cases began occurring in Korea in mid-February 2020. Since the end of February, the rate of infection has increased greatly due to mass (herd) infection within religious groups and nursing homes in the Daegu and Gyeongbuk regions. This mass infection has increased the number of infected people more rapidly than was initially expected; the epidemic model based on existing studies had predicted a much lower infection rate and faster recovery., Methods: The present study evaluated rapid infection spread by mass infection in Korea and the high mortality rate for the elderly and those with underlying diseases through the Susceptible-Exposed-Infected-Recovered-Dead (SEIRD) model., Results: The present study demonstrated early infection peak occurrence (-6.3 days for Daegu and -5.3 days for Gyeongbuk) and slow recovery trend (= -1,486.6 persons for Daegu and -223.7 persons for Gyeongbuk) between the actual and the epidemic model for a mass infection region compared to a normal infection region., Conclusion: The analysis of the time difference between infection and recovery can help predict the epidemic peak due to mass (or normal) infection and can also be used as a time index to prepare medical resources., Competing Interests: The authors have no potential conflicts of interest to disclose., (© 2020 The Korean Academy of Medical Sciences.)

Strain MEBiC09520 T , which was isolated from a tidal sediment in Incheon, Korea, is a pale yellow, rod-shaped bacterium, cells of which are 0.4-0.5 µm in width and 1.5-2 µm in length. Strain MEBiC09520 T shared 95.17 and 92.57% 16S rRNA gene sequence similarity with Emcibacter nanhaiensis and E. congregatus , respectively. It grew optimally at pH 6.0, at 55 °C and with 2.5-3.5% (w/v) NaCl. Its polar lipid components included phosphatidylethanolamine (PE), diphosphatidylglycerol (DPG), phosphatidylglycerol (PG), an unidentified phospholipid (PL), three unidentified aminolipids (ALs) and two unidentified lipids (L). The fatty acids C 16:0 , C 19:0 cyclo ω8 c , C 14:0 2-OH and summed feature 8 (C 18:1 ω7 c and/or C 18:1 ω6 c ) were predominantly present in its cell wall. Strain MEBiC09520 T was thermophilic, while E. nanhaiensis and E. congregatus were mesophilic. Although E. nanhaiensis showed no nitrate reduction activity, MEBiC09520 T and E. congregatus showed a positive reaction. These strains differed in carbohydrate utilization. In particular, E. congregatus was able to thrive on various carbohydrate substrates as compared to the other strains. The average nucleotide identity value was 69.92% between strain MEBiC09520 T and E. congregatus ZYL T , 70.38% between E. congregatus ZYL T and E. nanhaiensis HTCJW17 T , and 72.83% between strain MEBiC09520 and E. nanhaiensis HTCJW17 T . Considering these differences, strain MEBiC09520 T (=KCCM 43320 T =MCCC 1K03920 T ) is suggested to represent and novel species of a new genus, Luteithermobacter gelatinilyticus gen. nov., sp. nov., and E. congregatus should be reclassified as Paremcibacter congregatus gen. nov., comb. nov.

Glass is a well-known non-conductive material that has many useful properties, and considerable research has been conducted into making circuits on glass. Many deposition techniques have been studied, and laser-induced chemical liquid phase deposition (LCLD) is a well-known and cost-effective method for rapid prototyping of copper deposition on glass. However, the deposition results from the LCLD method on the surface of glass, which shows an issue in its detachment from the substrates because of the relatively low adhesion between deposited copper and the nontreated glass surface. This problem undermines the usability of deposited glass in industrial applications. In this study, the laser-induced backside wet etching (LIBWE) method was performed as a preceding process to fabricate microchannels, which were filled with copper by LCLD. Additional durable copper wire was produced as a result of the enhanced adhesion between the glass and the deposited copper. The adhesion was enhanced by a rough surface and metal layer, which are characteristics of LIBWE machining. Furthermore, the proposed method is expected to broaden the use of deposited glass in industrial applications, such as in stacked or covered multilayer structures with built-in copper wires, because the inserted copper can be physically protected by the microstructures.

Methanotrophs with soluble methane monooxygenase (sMMO) show high potential for various ecological and biotechnological applications. Here, we developed a high throughput method to identify sMMO-producing microbes by integrating droplet microfluidics and a genetic circuit-based biosensor system. sMMO-producers and sensor cells were encapsulated in monodispersed droplets with benzene as the substrate and incubated for 5 h. The sensor cells were analyzed as the reporter for phenol-sensitive transcription activation of fluorescence. Various combinations of methanotrophs and biosensor cells were investigated to optimize the performance of our droplet-integrated transcriptional factor biosensor system. As a result, the conditions to ensure sMMO activity to convert the starting material, benzene, into phenol, were determined. The biosensor signals were sensitive and quantitative under optimal conditions, showing that phenol is metabolically stable within both cell species and accumulates in picoliter-sized droplets, and the biosensor cells are healthy enough to respond quantitatively to the phenol produced. These results show that our system would be useful for rapid evaluation of phenotypes of methanotrophs showing sMMO activity, while minimizing the necessity of time-consuming cultivation and enzyme preparation, which are required for conventional analysis of sMMO activity., (Copyright © 2020 Lee, Baek, Kim, Kwon, Rha, Yeom, Kim, Lee, Kim and Lee.)

Genetic circuit-based biosensors have emerged as an effective analytical tool in synthetic biology; these biosensors can be applied to high-throughput screening of new biocatalysts and metabolic pathways. Sigma 54 (σ 54 )-dependent transcription factor (TF) can be a valuable component of these biosensors owing to its intrinsic silent property compared to most of the housekeeping sigma 70 (σ 70 ) TFs. Here, we show that these unique characteristics of σ 54 -dependent TFs can be used to control the host cell state to be more appropriate for high-throughput screening. The acclimation of cell state was achieved by using guanosine (penta)tetraphosphate ((p)ppGpp)-related genes (relA, spoT) and nutrient conditions, to link the σ 54 TF-based reporter expression with the target enzyme activity. By controlling stringent programmed responses and optimizing assay conditions, catalytically improved tyrosine phenol lyase (TPL) enzymes were successfully obtained using a σ 54 -dependent DmpR as the TF component, demonstrating the practical feasibility of this biosensor. This combinatorial strategy of biosensors using σ factor-dependent TFs will allow for more effective high-throughput enzyme engineering with broad applicability.

A strictly anaerobic, dissimilatory Fe(III)-reducing hyperthermophilic archaeon, designated as strain IOH1 T , was isolated from a new deep-sea hydrothermal vent (Onnuri Vent Field) area in the Central Indian Ocean ridge. Strain IOH1 T showed > 99% 16S rRNA gene sequence similarity with Thermococcus celericrescens TS2 T (99.4%) and T. siculi DSM 12349 T (99.2%). Additional three species T. barossii SHCK-94 T (99.0%), T. celer Vu13 T (98.8%), and T. piezophilus (98.6%) showed > 98.6% of 16S rRNA gene sequence similarity, however, the maximum OrthoANI value is 89.8% for the genome of T. celericrescens TS2 T . Strain IOH1 T cells are coccoid, 1.2-1.8 μm in diameter, and motile by flagella. Growth was at 70-82°C (optimum 80°C), pH 5.4-8.0 (optimum pH 6.0) with 2-4% (optimum 3%) NaCl. Growth of strain IOH1T was enhanced by starch, pyruvate, D(+)-maltose and maltodextrin as a carbon sources, and elemental sulfur as an electron acceptor; clearly different from those of related species T. celecrescens DSM 17994 T and T. siculi DSM 12349 T . Strain IOH1 T , T. celercrescence DSM 17994 T , and T. siculi DSM 12349 T reduced soluble Fe(III)-citrate present in the medium, whereas the amount of total cellular proteins increased with the concomitant accumulation of Fe(II). We determined a circular chromosome of 2,234 kb with an extra-chromosomal archaeal plasmid, pTI1, of 7.7 kb and predicted 2,425 genes. The DNA G + C content was 54.9 mol%. Based on physiological properties, phylogenetic, and genome analysis, we proposed that strain IOH1 T (= KCTC 15844 T = JCM 39077 T ) is assigned to a new species in the genus Thermococcus and named Thermococcus indicus sp. nov.

An anaerobic, rod-shaped, mesophilic, chemolithoautotrophic, sulfate-reducing bacterial strain IOR2 T was isolated from a newly found deep-sea hydrothermal vent (OVF, Onnuri Vent Field) area in the central Indian Ocean ridge (11°24'88″ S 66°25'42″ E, 2021 m water depth). The 16S rRNA gene sequence analysis revealed that the strain IOR2 T was most closely related to Desulfovibrio senegalensis BLaC1 T (96.7%). However, it showed low similarity with the members of the family Desulfovibrionaceae, such as Desulfovibrio tunisiensis RB22 T (94.0%), D. brasiliensis LVform1 T (93.9%), D. halophilus DSM 5663 T (93.7%), and Pseudodesulfovibrio aespoeensis Aspo-2 T (93.2%). The strain IOR2 T could grow at 23-42°C (optimum 37°C), pH 5.0-8.0 (optimum pH 7.0) and with 0.5-6.5% (optimum 3.0%) NaCl. The strain could use lactate, pyruvate, H 2 , and glycerol as electron donors and sulfate, thiosulfate, and sulfite as electron acceptors. The major fatty acids of the strain IOR2 T were iso-C 15:0 , iso-C 17:0 , ante-iso-C 15:0 , and summed feature 9 (C 16:0 methyl/iso-C 17:1 ω9c). Both the strains IOR2 T and BLaC1 T could grow with CO 2 and H 2 as the sole sources of carbon and energy, respectively. Genomic evidence for the Wood-Ljungdahl pathway in both the strains reflects chemolithoautotrophic growth. The DNA G + C content of the strain IOR2 T and BLaC1 T was 58.1-60.5 mol%. Based on the results of the phylogenetic and physiologic studies, Paradesulfovibrio onnuriensis gen. nov., sp. nov. with the type strain IOR2 T (= KCTC 15845 T = MCCC 1K04559 T ) was proposed to be a member of the family Desulfovibrionaceae. We have also proposed the reclassification of D. senegalensis as Paradesulfovibrio senegalensis comb. nov.

An important function in the future healthcare system involves measuring a patient's vital signs, transmitting the measured vital signs to a smart device or a management server, analyzing it in real-time, and informing the patient or medical staff. Internet of Medical Things (IoMT) incorporates information technology (IT) into patient monitoring device (PMD) and is developing traditional measurement devices into healthcare information systems. In the study, a portable ubiquitous-Vital (u-Vital) system is developed and consists of a Vital Block (VB), a small PMD, and Vital Sign Server (VSS), which stores and manages measured vital signs. Specifically, VBs collect a patient's electrocardiogram (ECG), blood oxygen saturation (SpO2), non-invasive blood pressure (NiBP), body temperature (BT) in real-time, and the collected vital signs are transmitted to a VSS via wireless protocols such as WiFi and Bluetooth. Additionally, an efficient R-point detection algorithm was also proposed for real-time processing and long-term ECG analysis. Experiments demonstrated the effectiveness of measurement, transmission, and analysis of vital signs in the proposed portable u-Vital system., Competing Interests: The authors declare no conflict of interest.

Bioconversion of C1 chemicals such as methane and methanol into higher carbon-chain chemicals has been widely studied. Methanol oxidation catalyzed by methanol dehydrogenase (Mdh) is one of the key steps in methanol utilization in bacterial methylotrophy. In bacteria, few NAD + -dependent Mdhs have been reported that convert methanol to formaldehyde. In this study, an uncharacterized Mdh gene from Lysinibacillus xylanilyticus (Lxmdh) was cloned and expressed in Escherichia coli . The maximum alcohol oxidation activity of the recombinant enzyme was observed at pH 9.5 and 55°C in the presence of 10 mM Mg 2+ . To improve oxidation activity, rational approach-based, site-directed mutagenesis of 16 residues in the putative active site and NAD + -binding region was performed. The mutations S101V, T141S, and A164F improved the enzyme's specific activity toward methanol compared to that of the wild-type enzyme. These mutants show a slightly higher turnover rate than that of wild-type, although their K M values were increased compared to that of wild-type. Consequently, according the kinetic results, S101, T141, and A164 positions may related to the catalytic activity in the active site for methanol dehydrogenation. It should be further studied other mutant variants with high activity for methanol. In conclusion, we characterized a new Lxmdh and its variants that may be potentially useful for the development of synthetic methylotrophy in the future., (Copyright © 2020 Lee, Park, Oh, Lee, Kwon, Kim, Choi, Rha, Lee, Lee, Sung, Yeom and Lee.)

A mesophilic, straight-rod-shaped, non-flagellated bacterium, designated MEBiC05444 T , was isolated from a marine sponge collected from Chuuk lagoon, Federated States of Micronesia. The strain was Gram-negative, catalase- and oxidase-positive, and facultative anaerobic. The isolate aerobically grew at 8-38 °C (optimum, 24-32 °C), pH 4.0-10.0 (pH 7.0-7.5) with an absolute requirement for Na + up to 6 % (w/v) NaCl (2 %). Phylogenetic analyses based on 16S rRNA gene sequences revealed that MEBiC05444 T belonged to the family Shewanellaceae , within the class Gammaproteobacteria. Strain MEBiC05444 T showed highest 16S rRNA gene sequence similarity to Parashewanella curva C51 T , followed by [ Shewanella ] irciniae UST040317-058 T and Parashewanella spongiae HJ039 T (98.9 %, 97.2 and 95.7 %, respectively). In the phylogenetic tree based on the 16S rRNA gene sequences, MEBiC05444 T formed a cluster with P. curva C51 T , but the average nucleotide identity value between the two strains was 82 %, thus confirming their separation at species level. The major fatty acids were iso-C 15 : 0 (19.7 %), summed feature 3 (composed of C 16 : 1  ω7 c and/or C 16 : 1 ω6 c ; 16.1 %) and C 17 : 1 ω8 c (10.2 %). The only detected respiratory quinone was ubiquinone Q-8. The major polar lipids were phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, three unidentified aminoglycolipids, two unidentified glycolipids, an unidentified aminoglycophospholipid and an unidentified lipid. The genomic DNA G+C content of strain MEBiC05444 T was 40.8 mol%. Based on the results of polyphasic analysis, the strain represents a novel species of the genus Parashewanella , distinct from P. curva C51 T , [ Shewanella ] irciniae UST040317-058 T and P. spongiae HJ039 T for which the name Parashewanellatropica sp. nov. is proposed with type strain MEBiC05444 T (=KCCM 43304 T =JCM 16653 T ).

While various bioremediation techniques have been widely used at oil spill sites, the in situ efficiency of such techniques on recovering the benthic communities in intertidal areas has not been quantified. Here, the performance of several bioremediation tools such as emulsifiers, multi-enzyme liquid (MEL), microbes, and rice-straw was evaluated by a 90-days semi-field experiment, particularly targeting recovery of benthic community. Temporal efficiency in the removal of sedimentary total petroleum hydrocarbons (TPH), reduction of residual toxicity, and recovery of bacterial diversity, microalgal growth, and benthic production was comprehensively determined. Concentrations of TPH and amphipod mortality for all treatments rapidly decreased within the first 10 days. In addition, the density of bacteria and microphytobenthos generally increased over time for all treatments, indicating recovery in the benthic community health. However, the recovery of some nitrifying bacteria, such as the class Nitrospinia (which are sensitive to oil components) remained incomplete (13-56%) during 90 days. Combination of microbe treatments showed rapid and effective for recovering the benthic community, but after 90 days, all treatments showed high recovery efficiency. Of consideration, the "no action" treatment showed a similar level of recovery to those of microbe and MEL treatments, indicating that the natural recovery process could prevail in certain situations., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Strain IMCC1322 was isolated from a surface water sample from the East Sea of Korea. Based on 16S rRNA analysis, IMCC1322 was found to belong to the OCS28 sub-clade of SAR116. The cells appeared as short vibrioids in logarithmic-phase culture, and elongated spirals during incubation with mitomycin or in aged culture. Growth characteristics of strain IMCC1322 were further evaluated based on genomic information; proteorhodopsin (PR), carbon monoxide dehydrogenase, and dimethylsulfoniopropionate (DMSP)-utilizing enzymes. IMCC1322 PR was characterized as a functional retinylidene protein that acts as a light-driven proton pump in the cytoplasmic membrane. However, the PR-dependent phototrophic potential of strain IMCC1322 was only observed under CO-inhibited and nutrient-limited culture conditions. A DMSP-enhanced growth response was observed in addition to cultures grown on C 1 compounds like methanol, formate, and methane sulfonate. Strain IMCC1322 cultivation analysis revealed biogeochemical processes characteristic of the SAR116 group, a dominant member of the microbial community in euphotic regions of the ocean. The polyphasic taxonomy of strain IMCC1322 is given as Candidatus Puniceispirillum marinum, and was confirmed by chemotaxonomic tests, in addition to 16S rRNA phylogeny and cultivation analyses.

A Gram-stain-negative oval-rod-shaped, spore-forming anaerobic bacterium, designated as strain MCWD5 T , was isolated from sediment of a salt pond in the Republic of Korea (35° 7' 18″ N 126° 19' 4″ E). The 16S rRNA gene sequence analysis revealed that strain MCWD5 T had low similarity values to members in the family Lachnospiraceae, such as Robinsoniella peoriensis PPC31 T (94.8 %), Ruminococcusgauvreauii CCRI-16110 T (94.2 %) and Lachnotalea glycerini DLD10 T (94.0 %), and its phylogenetic position is unstable. The strain could grow at 20-42 °C (optimum, 38-42 °C), pH 5.5-10.0 (pH 7.0) and with 0-6 % (2.0 %) NaCl. Strain MCWD5 T could not use nitrate, nitrite, sulfate or sulfite as electron acceptors. The strain could utilize various carbohydrates, such as arabinose, cellobiose, glucose, etc., and polymers such as pectin and starch. The major fatty acids of strain MCWD5 T were C14 : 0, C16 : 0, C16 : 1ω7c, C18 : 1ω7c DMA and summed feature 8 (C17 : 1ω8c and/or C17 : 2), which was clearly different from those of related genera. The major polar lipids were diphosphatidyglycerol, phosphatidyglycerol and an unknown phospholipid. Based on the results of phylogenetic, physiologic and chemotaxonomic studies, Anaerosacchariphilus polymeriproducens gen. nov., sp. nov. with the type strain MCWD5 T (=KCTC 15595 T =DSM 105757 T ) is proposed in the family Lachnospiraceae.