Impact of IL-1ß on lung pathology caused by Mycobacterium abscessus infection and its association with IL-17 production.

Mycobacterium abscessus (MAB), a non-tuberculous mycobacterium (NTM), causes chronic pulmonary inflammation in humans. The NLRP3 inflammasome is a multi-protein complex that triggers IL-1ß maturation and pyroptosis through the cleavage of caspase-1. In this study, we investigated the roles of NLRP3 and IL-1ß in the host's defense against MAB. The IL-1ß production by MAB was completely abolished in NLRP3, but not NLRC4, deficient macrophages. The NLRP3 inflammasome components, which are ASC and caspase-1 were also found to be essential for IL-1ß production in response to MAB. NLRP3 and IL-1ß deficiency did not affect the intracellular growth of MAB in macrophages, and the bacterial burden in lungs of NLRP3- and IL-1ß-deficient mice was also comparable to the burden observed in WT mice. In contrast, IL-1ß deficiency ameliorated lung pathology in MAB-infected mice. Notably, the lung homogenates of IL-1ß-deficient mice had reduced levels of IL-17, but not IFN-γ and IL-4 when compared with WT counterparts. Furthermore, in vitro co-culture analysis showed that IL-1ß signaling was essential for IL-17 production in response to MAB. Finally, we observed that the anti-IL-17 antibody administration moderately mitigated MAB-induced lung pathology. These findings indicated that IL-1ß production contribute to MAB-induced lung pathology via the elevation of IL-17 production.

Didymella acutilobae sp. nov. Causing Leaf Spot and Stem Rot in Angelica acutiloba.

During disease surveys of Angelica acutiloba plants in Korea, leaf spot symptoms were observed in a field in Andong in July 2019, and stem rot symptoms in vinyl greenhouses in Yangpyeong in April 2020. Incidence of leaf spot and stem rot of the plants ranged from 10 to 20% and 5 to 30%, respectively. Morphological and cultural characteristics of fungal isolates from the leaf spot and stem rot symptoms fitted into those of the genus Phoma. Molecular phylogenetic analyses of two single-spore isolates from the symptoms using concatenated sequences of LSU, ITS, TUB2, and RPB2 genes authenticated an independent cluster from other Didymella (anamorph: Phoma) species. Moreover, the isolates showed different morphological and cultural characteristics in comparison to closely related Didymella species. These discoveries confirmed the novelty of the isolates. Pathogenicity of the novel Didymella species isolates was substantiated on leaves and stems of A. acutiloba through artificial inoculation. Thus, this study reveals that Didymella acutilobae sp. nov. causes leaf spot and stem rot in Angelica acutiloba.

Wide-field three-dimensional photoacoustic/ultrasound scanner using a two-dimensional matrix transducer array.

Two-dimensional matrix transducer arrays are the most appropriate imaging probes for acquiring dual-modal 3D photoacoustic (PA)/ultrasound (US) images. However, they have small footprints which limit the field-of-view (FOV) to less than 10 mm × 10 mm and degrade the spatial resolution. In this study, we demonstrate a dual-modal PA and US imaging system (using a 2D matrix transducer array and a motorized 2D scanning system) to enlarge the FOV of volumetric images. Multiple PA volumes were merged to form a wide-field image of approximately 45 mm × 45 mm. In vivo imaging was demonstrated using rat sentinel lymph nodes (SLNs) and bladders stained with methylene blue. We believe that this volumetric PA/US imaging technique with a 2D matrix transducer array can be a useful tool for narrow-field real-time monitoring and wide-field imaging of various preclinical and clinical studies.

Didymella gigantis sp. nov. Causing Leaf Spot in Korean Angelica.

During a disease survey in October 2019, leaf spot symptoms with a yellow halo were observed on Korean angelica (Anglica gigas) plants grown in fields in Pyeongchang, Gangwon Province, Korea. Incidence of diseased leaves of the plants in the investigated fields ranged from 10% to 60%. Morphological and cultural characteristics of two single-spore isolates from the leaf lesions indicated that they belonged to the genus Didymella. Molecular phylogenetic analyses using combined sequences of LSU, ITS, TUB2, and RPB2 regions showed distinct clustering of the isolates from other Didymella species. In addition, the morphological and cultural characteristics of the isolates were somewhat different from those of closely related Didymella spp. Therefore, the novelty of the isolates was proved based on the investigations. Pathogenicity of the novel Didymella species isolates was confirmed on leaves of Korean angelica plants via artificial inoculation. This study reveals that Didymella gigantis sp. nov. causes leaf spot in Korean angelica.

First report of tomato pith necrosis caused by Pseudomonas mediterranea in South Korea.

Tomato (Solanum lycopersicum) is one of the most economically important vegetables worldwide, and its production is directly affected by several bacterial diseases (Singh et al., 2017). During a disease survey in 2020, pith necrosis-like symptoms, commonly caused by Pseudomonas spp., were observed in two commercial greenhouses in PyeongChang and Gyeongju, South Korea. Disease incidence ranged from 8 to 10%, and infected plants showed wilt symptoms, brown stem discoloration, leaf blight, and corrugated pith tissues (eXtra Fig. 1). Symptomatic stem tissues were surface disinfected, cut into small pieces, and macerated in sterile water. The resulting suspension was spread on nutrient agar, and incubated at 28°C. The dominant bacterial colony types were round, mucoid, and frequently produced yellow to brown pigments. Four bacterial colonies (CPB20664 - CPB20667), each from a different diseased plant, were selected for further study. All isolates were Gram-negative and did not produce fluorescent pigments on King's B medium. Biochemical profiles of the isolates were determined by the API20NE (Biomerieux, Durhan, NC, USA) and LOPAT test (eXtra Table 1). The bacterial isolates were further identified by PCR amplification of partial 16S rRNA, gyrB, and rpoD genes using primers 27F/1492R, UP-1E/AprU, and 70F/70R, respectively (Lane 1991, Yamamoto et al., 2000). The resulting sequences were deposited in GenBank under accession numbers MW602997 to MW603000 for 16S rRNA, MW602987 to MW602990 for gyrB, and MW602991 to MW602994 for rpoD. These sequences exhibited 99-100% nucleotide similarities with multiple Pseudomonas mediterranea sequences in Genbank. Additionally, the isolates were subjected to PCR assays using the P. mediterranea specific primers PC5/1-PC5/2 and the P. corrugata specific primers PC1/1-PC1/2 (Catara et al., 2002). All isolates produced a specific 600-bp band with the P. mediterranea primers, but did not produce any bands with the P. corrugata specific primers. The PCR amplicons were sequenced and BLAST queried against GenBank database. All isolates shared 100% identity with the type strain P. mediterranea DSM 16733 (acc No. LT629790.1). These results indicated that the bacteria isolated from the tomato plants with pith necrosis were P. mediterranea. Pathogenicity tests were conducted on 2-week-old tomato seedlings (cv. Yekwang) by wound inoculations. Single colonies were picked up using sterile toothpicks, and the stems of tomato seedlings were stabbed below the second leaves. As a negative control, a sterile toothpick was dipped in sterile water and used in the same manner. After inoculation, the plants were kept in a humidity box for 48 h, then moved to a plant growth room. After 15 days, light brown lesions had developed at the stab sites, and pith necrosis and slight wilting of plants were observed at 30 days (eXtra Fig. 1). Control plants remained asymptomatic. P. mediterranea was re-isolated from infected plants, fulfilling Koch's postulates. Five species of Pseudomonas are known to cause tomato pith necrosis (Alippi and Lopez, 2010, Cañizares and García-Pedrajas., 2015, Ruan et al., 2018) including P. corrugata previously reported from Korea (Choi and Han, 2004). This is believed to be the first report of P. mediterranea as the cause of tomato pith necrosis in Korea. Tomato pith necrosis disease reduces the quality and yield of tomato production and appropriate management strategies should be investigated to control this disease.

Engineered extracellular vesicle-based sonotheranostics for dual stimuli-sensitive drug release and photoacoustic imaging-guided chemo-sonodynamic cancer therapy.

Sonodynamic therapy has shown promise as an effective alternative to conventional photodynamic therapy owing to its ability to treat deep-seated tumors. However, the development of stimuli-responsive sonosensitizers with high biocompatibility faces a significant challenge. Methods: In this study, we developed dual stimuli-responsive sonosensitizers with desirable biosafety using extracellular vesicles (EVs), a class of naturally occurring nanoparticles. Indocyanine green (ICG), which functions as both a sonosensitizer and photoacoustic (PA) imaging agent, was loaded into EVs, together with paclitaxel (PTX) and sodium bicarbonate (SBC), to achieve pH-responsive PA imaging-guided chemo-sonodynamic combination therapy. Results: The EVs significantly improved the cellular uptake of ICG, thus triggering enhanced sonodynamic effects in breast cancer cells. SBC-, ICG-, and PTX-loaded EV [SBC-EV(ICG/PTX)] efficiently released the PTX in response to acidic pH in the endo/lysosomes because CO2 bubbles generated from the SBC caused the EV membranes to burst. The drug release was further facilitated by ultrasound (US) treatment, demonstrating dual pH/US-responsive drug release. The ICG- and PTX-loaded EVs exhibited efficient anticancer activity against breast tumor cells owing to the combination of chemo-sonodynamic therapy. High-resolution PA imaging visualized the preferential tumor accumulation of SBC-EV(ICG/PTX) in tumor-bearing mice. Notably, a single intravenous injection of SBC-EV(ICG/PTX) with US irradiation significantly suppressed tumor growth in mice without systemic toxicity. Conclusions: Our findings demonstrate that dual stimuli-responsive SBC-EV(ICG/PTX) are promising sonotheranostic nanoplatforms for safe and efficient chemo-sonodynamic combination cancer therapy and photoacoustic imaging.

Erratum: "New contrast agents for photoacoustic imaging and theranostics: Recent 5-year overview on phthalocyanine/naphthalocyanine-based nanoparticles" [APL Bioeng. 5, 031510 (2021)].

[This corrects the article DOI: 10.1063/5.0047660.].

New contrast agents for photoacoustic imaging and theranostics: Recent 5-year overview on phthalocyanine/naphthalocyanine-based nanoparticles.

The phthalocyanine (Pc) and naphthalocyanine (Nc) nanoagents have drawn much attention as contrast agents for photoacoustic (PA) imaging due to their large extinction coefficients and long absorption wavelengths in the near-infrared region. Many investigations have been conducted to enhance Pc/Ncs' photophysical properties and address their poor solubility in an aqueous solution. Many diverse strategies have been adopted, including centric metal chelation, structure modification, and peripheral substitution. This review highlights recent advances on Pc/Nc-based PA agents and their extended use for multiplexed biomedical imaging, multimodal diagnostic imaging, and image-guided phototherapy.

Water extract of Artemisia scoparia Waldst. & Kitam suppresses LPS-induced cytokine production and NLRP3 inflammasome activation in macrophages and alleviates carrageenan-induced acute inflammation in mice.

ETHNOPHARMACOLOGICAL RELEVANCE: Artemisia scoparia Waldst. & Kitam (A. scoparia) is a perennial herbal plant that is widely used as a folk remedy in Asian countries. Several studies have demonstrated that A. scoparia has various physiological effects, including anti-inflammation, anti-hypertension, anti-obesity, anti-hepatotoxicity, and anti-oxidant effects. AIM OF THE STUDY: The objective of the present study was to examine the anti-inflammatory effects of water extract of A. scoparia (WAS). MATERIALS AND METHODS: Murine bone marrow-derived macrophages (BMDMs), human monocyte THP-1 and murine fibroblast 3T3-L1 cells were used for the in vitro experiments. Cell viability and cytokine production were determined by the MTT assay and ELISA, respectively. RT-PCR was performed to determine iNOS gene expression and the Griess reaction was used to measure nitrite levels. iNOS protein expression, activation of NF-κB and MAPKs, and cleavage of caspase-1 and IL-1ß were determined by Western blot analysis. A carrageenan-induced mouse model of acute inflammation was used in the in vivo experiments. RESULTS: Pretreatment with WAS concentration-dependently suppressed gene expression and IL-6, TNF-α, CXCL1 and iNOS protein levels in BMDMs stimulated with LPS. In addition, pretreatment with WAS inhibited LPS-induced production of IL-6 and TNF-α in THP-1 cells and CXCL1 in 3T3-L1. Furthermore, LPS induced phosphorylation of p65 in BMDMs, and this induction was dramatically suppressed by WAS pretreatment. We further investigated whether WAS regulates activation of the NLRP3 inflammasome, which is known to be essential for IL-1ß processing. WAS inhibited the production of IL-1ß, but not IL-6, in response to adenosine triphosphate (ATP) and monosodium uric acid (MSU) crystals in LPS-primed BMDMs. Cleavage of caspase-1 and IL-1ß was also reduced by WAS. We finally evaluated the in vivo anti-inflammatory effects of WAS in a mouse model of carrageenan-induced acute inflammation. Subcutaneous administration of WAS reduced production of the inflammatory cytokines IL-6, TNF-α, CXCL1, and IL-1ß. Recruitment of immune cells, mostly neutrophils, was also reduced by administration of WAS. Infiltration of inflammatory cells and edema in the submucosa of air pouch tissues were markedly improved in the WAS-treated groups. CONCLUSIONS: Our results indicate that WAS possesses potent anti-inflammatory properties. These findings suggest that A. scoparia is a candidate functional food targeting several inflammatory diseases.

Complete Genome Sequence of Bacillus amyloliquefaciens subsp. plantarum CC178, a Phyllosphere Bacterium Antagonistic to Plant Pathogenic Fungi.

Bacillus amyloliquefaciens subsp. plantarum strain CC178 is a phyllosphere bacterium with antagonistic activity against a wide range of plant fungal pathogens. The genome of strain CC178 is 3,916,828 bp in size and harbors 3,972 genes. Six giant gene clusters are dedicated to the nonribosomal synthesis of antimicrobial polypeptides and polyketides.

Rudaeicoccus suwonensis gen. nov., sp. nov., an actinobacterium isolated from the epidermal tissue of a root of a Phalaenopsis orchid.

A bacterial strain, designated HOR6-4(T), was isolated from the epidermal tissue of a root of a Phalaenopsis orchid. Strain HOR6-4(T) was characterized using a polyphasic approach. The cells were aerobic, Gram-stain-positive, non-motile coccoids. Phylogenetic analysis of its 16S rRNA gene sequence revealed a clear affiliation with the family Dermacoccaceae. Strain HOR6-4(T) showed relatively low 16S rRNA gene sequence similarity (below 95.9 %) to type strains of species with validly published names, showing the highest sequence similarities to the type strains of Flexivirga alba (95.8 %) and Yimella lutea (95.5 %). The DNA G+C content of strain HOR6-4(T) was 64.7 mol%. Strain HOR6-4(T) had anteiso-C17 : 0 (19.3 %), 10-methyl C18 : 0 (tuberculostearic acid; 13.5 %) and 10-methyl C17 : 0 (11.7 %) as the major fatty acids and contained MK-8(H4) and MK-8(H6) as the predominant quinones. Polar lipids were diphosphatidylglycerol, phosphatidylinositol, three aminophospholipids, two phospholipids and an aminolipid. The peptidoglycan type was A4α, with an L-Lys-L-Thr-D-Glu interpeptide bridge with a glycine residue bound to the alpha-carboxyl group of D-Glu in position 2 of the peptide subunit. Mycolic acids were not present. On the basis of comparative analysis of phenotypic and phylogenetic traits, strain HOR6-4(T) is considered to represent a novel species in a new genus in the family Dermacoccaceae, for which the name Rudaeicoccus suwonensis gen. nov., sp. nov. is proposed. The type strain of Rudaeicoccus suwonensis is HOR6-4(T) ( = KACC 12637(T)  = DSM 19560(T)).

Lysinimonas soli gen. nov., sp. nov., isolated from soil, and reclassification of Leifsonia kribbensis Dastager et al. 2009 as Lysinimonas kribbensis sp. nov., comb. nov.

A Gram-stain-positive, non-motile rod, designated strain SGM3-12(T), was isolated from paddy soil in Suwon, Republic of Korea. 16S rRNA gene sequence analysis revealed that the strain represented a novel member of the family Microbacteriaceae. The nearest phylogenetic neighbour was Leifsonia kribbensis MSL-13(T) (97.4 % 16S rRNA gene sequence similarity). Strain SGM3-12(T) and Leifsonia kribbensis MSL-13(T) formed a distinct cluster within the family Microbacteriaceae. Strain SGM3-12(T) contained MK-12(H2) and MK-11(H2) as the predominant menaquinones with moderate amounts of MK-12 and MK-11; anteiso-C15 : 0 and iso-C16 : 0 as the major cellular fatty acids (>10 % of total); and diphosphatidylglycerol, phosphatidylglycerol and unidentified glycolipids as the polar lipids. The peptidoglycan type of the isolate was B1δ with L-Lys as the diagnostic cell-wall diamino acid. On the basis of these results, strain SGM3-12(T) represents a novel species within a new genus, for which the name Lysinimonas soli gen. nov., sp. nov. is proposed (the type strain of the type species is SGM3-12(T) = KACC 13362(T) = NBRC 107106(T)). It is also proposed that Leifsonia kribbensis be transferred to this genus as Lysinimonas kribbensis comb. nov. (the type strain is MSL-13(T) = DSM 19272(T) = JCM 16015(T) = KACC 21108(T) = KCTC 19267(T)).

Draft genome sequence of the biocontrol bacterium Bacillus amyloliquefaciens strain M27.

Bacillus amyloliquefaciens strain M27 is a biocontrol agent with antagonistic activities against a wide range of fungal pathogens. Here we present the 3.86-Mb draft genome sequence of the bacterium with the aims of providing insights into the genomic basis of its antifungal mechanism and facilitating its application in the biocontrol of plant diseases.

Niabella soli sp. nov., isolated from soil from Jeju Island, Korea.

A dark yellow-coloured bacterium, JS13-8(T), was isolated from a soil sample from Jeju Island, Republic of Korea. The cells were aerobic, Gram-negative, non-motile, short rods (0.5-0.7 x 0.8-1.4 microm). Growth occurred at 15-35 degrees C (optimally at 30 degrees C), at pH 5.0-8.0 (optimally at pH 6.0-7.0) and at 0-1 % NaCl (w/v). Flexirubin pigment was produced. A phylogenetic analysis based on 16S rRNA gene sequences revealed that strain JS13-8(T) was closely related to Niabella aurantiaca KACC 11698(T) (95.0 % sequence similarity). The major respiratory quinone system was MK-7 and the predominant cellular fatty acids were iso-C(15 : 0), iso-C(15 : 1) G, iso-C(17 : 0) 3-OH and summed feature 3. The DNA G+C content was 45 mol%. On the basis of the phylogenetic, physiological and chemotaxonomic data, strain JS13-8(T) represents a novel species of the genus Niabella, for which the name Niabella soli sp. nov. is proposed. The type strain is strain JS13-8(T) (=KACC 12604(T)=DSM 19437(T)).

Paludibacterium yongneupense gen. nov., sp. nov., isolated from a wetland, Yongneup, in Korea.

A cream-coloured bacterial strain, 5YN8-15(T), was isolated from a wetland, Yongneup, in the Inje region of the Republic of Korea. The bacterium was facultatively anaerobic, Gram-negative, motile with a single polar flagellum and curved-rod-shaped. Based on 16S rRNA gene sequence analysis, strain 5YN8-15(T) is a member of the Betaproteobacteria. Closely related taxa were Gulbenkiania mobilis E4FC31(T) (94.9% sequence similarity), Chromobacterium species (94.1-94.4%), Aquitalea magnusonii TRO-001DR8(T) (93.2%) and Aquaspirillum serpens IAM 13944(T) (92.5%). All other species with validly published names analysed showed sequence similarities of below 92%. Strain 5YN8-15(T) had ubiquinone 8 as the major isoprenoid quinone. The major fatty acids were summed feature 3 (C(16:1)omega7c and/or iso-C(15:0) 2-OH), C(16:0) and C(18:1)omega7c. The DNA G+C content was 63.0 mol%. Based on the data from the polyphasic study, strain 5YN8-15(T) represents a novel genus and species of the family Neisseriaceae, for which the name Paludibacterium yongneupense gen. nov., sp. nov. is proposed. The type strain is 5YN8-15(T) (=KACC 11601(T)=DSM 18731(T)).

Andreprevotia chitinilytica gen. nov., sp. nov., isolated from forest soil from Halla Mountain, Jeju Island, Korea.

A motile, Gram-negative, rod-shaped bacterium, designated strain JS11-7(T), was isolated from forest soil from Halla Mountain, Jeju Island, Korea. It grew optimally at 28 degrees C, pH 6-7 and in the presence of 0-2 % (w/v) NaCl. Phylogenetic analyses based on 16S rRNA gene sequences showed that strain JS11-7(T) belongs to the family Neisseriaceae, with Silvimonas terrae as its closest relative (95.0 % similarity). It contained Q-8 as the predominant quinone and summed feature 3 (comprising iso-C(15 : 0) 2-OH and/or C(16 : 1) omega 6c), C(16 : 0) and C(18 : 1) omega 7c as the major fatty acids. The DNA G+C content was 62 mol%. On the basis of phenotypic and genotypic characteristics and the results of 16S rRNA gene sequence analysis, strain JS11-7(T) represents a novel genus and species, for which the name Andreprevotia chitinilytica gen. nov., sp. nov. is proposed. The type species is Andreprevotia chitinilytica and the type strain is JS11-7(T) (=KACC 11608(T)=DSM 18519(T)).

Pedobacter suwonensis sp. nov., isolated from the rhizosphere of Chinese cabbage (Brassica campestris).

A mesophilic bacterium, strain 15-52(T), was isolated from the rhizosphere of Chinese cabbage (Brassica campestris). On the basis of phenotypic and genotypic characteristics, the bacterium was identified as representing a novel species belonging to the genus Pedobacter. The strain is non-flagellated, non-spore-forming and grows at temperatures in the range 1-37 degrees C. Physiological tests of the strain showed the presence of oxidase, catalase, protease (gelatin and casein hydrolysis), beta-glucosidase and beta-galactosidase activities. The highest levels of 16S rRNA gene sequence similarity were found with respect to Pedobacter roseus CL-GP80(T) (97.3 %) and Pedobacter sandarakinus DS-27(T) (97.2 %). A phylogenetic analysis based on 16S rRNA gene sequence data indicated that strain 15-52(T) is a member of the genus Pedobacter. DNA-DNA hybridization analysis revealed low levels of relatedness (<42.3 %) between the isolate and two phylogenetically related type strains, P. roseus KCCM 42272(T) and P. sandarakinus KCTC 12559(T). The DNA G+C content is 44.2 mol% and the predominant fatty acids are iso-C(15 : 0) (35.4 %), iso-C(15 : 0) 2-OH and/or C(16 : 1)omega7c (27.8 %) and iso-C(17 : 0) 3-OH (15.8 %). On the basis of these data, strain 15-52(T) represents a novel species of the genus Pedobacter, for which the name Pedobacter suwonensis sp. nov. is proposed. The type strain is 15-52(T) (=KACC 11317(T)=DSM 18130(T)).

Free radical scavengers from the medicinal mushroom Inonotus xeranticus and their proposed biogenesis.

New free radical scavengers, inoscavin D (1) and methylinoscavin D (2), were isolated from the methanolic extract of the fruiting bodies of Inonotus xeranticus (Hymenochaetaceae), along with the known compounds phelligridin D (3), 3,4-dihydroxybenzaldehyde (4), and 3,4-dihydroxybenzoic acid (5). Their structures were established by various spectroscopic analyses. Compounds 1 and 3 were proposed to be biosynthesized from the oxidative coupling of the precursor hispidin with 3,4-dihydroxybenzaldehyde and 3,4-dihydroxybenzoic acid, respectively. These compounds exhibited significant scavenging activity against the ABTS radical cation, and compounds 2 and 4 displayed moderate superoxide radical scavenging activity.

Diaporthin and Orthosporin from the Fruiting Body of Daldinia concentrica.

In our continuing study on the chemical constituents in the fruiting bodies of Daldinia concentrica, diaporthin and orthosporin were isolated. Their chemical structures were assigned based on various spectral studies. Diaporthin and orthosporin, phytotoxins previously found in Aspergillus ochraceus, were isolated from wood-rotting mushroom D. concentrica for the first time.