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Purpose: To compare eyelid blink characteristics between patients with ptosis and healthy controls using a smartphone camera. Methods: The ptosis group consisted of 20 senile aponeurotic ptosis patients with margin reflex distance1 ≤2.5 mm and the control group consisted of 10 healthy subjects without ptosis. The ptosis group was further divided into two groups based on an age cutoff of 70 years. Palpebral fissure height, levator function, margin reflex distance1, inter-blink interval, blink duration, blink rate, and blink velocity were measured and compared between the three groups based on photographs of the eyelids and videos of blinking taken with a smartphone camera. Results: The palpebral fissure height, levator function, margin reflex distance1, and blink velocity were lower in the ptosis groups than in the control group but these values did not differ between the two ptosis groups. The palpebral fissure height, levator function, and margin reflex distance1 were correlated with blink velocity. In the receiver operating characteristic (ROC) curve of blink velocity, the area under the receiver operating characteristic (AUROC) curve value was as high as 0.969 and the cut-off value was 32.36 mm/s. Conclusions: It is possible to analyze eyelid blink characteristics using a smartphone camera and the results confirmed that palpebral fissure height, levator function, margin reflex distance1, and blink velocity were lower in the senile aponeurotic ptosis group than in the healthy control group and were unaffected by age. Additionally, blink velocity is valuable for diagnosis of ptosis due to the correlation between the degree of ptosis, blink velocity, and the ROC curve of blink velocity.

Photorhabdus spp. has a high molecular weight Tc toxin with insecticidal activity. These toxins have been suggested as an alternative to BT toxin from Bacillus thuringiensis. Herein, we constructed a cosmid library with the genome of M1021 and screened the Escherichia coli clones showing insect toxicity by injecting each clone into Galleria mellonella larvae. In a total of 1020 clones, one clone with high insecticidal activity was selected and the nucleotide sequence of the cosmid of the clone was determined. In cosmid PtC28, a gene with 87% homology to the tccB gene of Photorhabdus temperata was found. Consequently, we have isolated the tccB gene cassette from the M1021 and expressed in E. coli expression vectors. The toxin was produced in the form of inclusion bodies but the denatured and refolded recombinant TccB showed strong mortality to the G. mellonella larvae.

Thrombosis is one of the main complications of endovascular treatment for unruptured intracranial aneurysms (UIA). This article reports the timing and initial location of thrombosis and prognosis after the use of tirofiban for performing coil embolization for UIA. This study retrospectively collected 1686 cases of intracranial aneurysms treated with coil embolization from January 2013 to February 2018. Ruptured cases were excluded. The presumed causes and timing of thrombosis, the response after tirofiban administration and the modified Rankin scale (mRS) score at 3 months were reviewed. Of the 26 patients 76% were female and middle cerebral artery and basilar artery aneurysms accounted for 7 cases. The initial location of thrombosis was related to the stent (n = 14, 53.8%) or coil (n = 12, 46.2%). Of the patients 19 (73.1%) developed thrombosis during the procedure, and 5 patients (19.2%) developed it within 1 day of the procedure. Median duration between the thrombotic procedure and initial thrombosis was 38.5 min, 12 patients were symptomatic but more than half completely recovered after using tirofiban. Good clinical outcome (mRS 0–2) was seen in 92.3%. In the subgroup analysis, median time from the first thrombotic procedure to initial thrombosis within 1 day was 38.0 min (stent-related group) and 35.0 min (coil-related group, p = 0.651). In most cases of embolization for UIA, thrombosis requiring the use of tirofiban occurs intraprocedurally or on the first day after the procedure. Careful observation of thrombosis during the procedure is important and tirofiban should be used for a better outcome even if the infarction progresses.

(E)-3-(3-(4-((3-Carbamoylbenzyl)oxy)-3-iodo-5-methoxyphenyl) acryloyl)benzamide (A6) was found to be a potent p300 inhibitor (IC50 = 870 nM) showing a similar binding mode to that of acetyl-CoA, a p300 substrate, and effective anti-fibrotic activity in both TGF-β1-stimulated lung fibroblast cells and bleomycin-induced in vivo lung fibrosis mice.

Strain KUDC5002T was isolated from soil sampled on the Dokdo Islands, Republic of Korea. This bacterial strain was Gram stain-positive, non-motile, rod-shaped, capable of growing at 25–37°C and pH 5.0–12.0, and showed optimal growth at 30 °C and pH 7.0–8.0. Strain KUDC5002T could be grown in tryptic soy broth containing less than 7.0 % NaCl (w/v). The cell width ranged from 0.5 to 0.6 µm and length ranged from 0.8 to 1.0 µm. Strain KUDC5002T was catalase- and oxidase-positive. Its genomic G+C content was 72.2 mol%. Its major fatty acids were C18 : 1 ω9c (17.3 %), iso-C16 : 0 (16.0 %) and iso-C17 : 0 (11.4 %). Phylogenetic analysis, based on 16S rRNA gene sequences, showed that strain KUDC5002T belongs to the genus Nocardioides and is most closely related to strain Nocardioides humi DCY24T (97.0 %). Based on its phenotypic, phylogenetic, genetic and chemotaxonomic features, strain KUDC5002T should be considered a novel species in the genus Nocardioides , for which we have proposed the name Nocardioides sambongensis sp. nov. The type strain is KUDC5002T (=KCTC 39855T=DSM 106604T).

BackgroundAdipic acid, a six-carbon platform chemical mainly used in nylon production, can be produced via reverse β-oxidation in microbial systems. The advantages posed byCorynebacterium glutamicumas a model cell factory for implementing the pathway include: (1) availability of genetic tools, (2) excretion of succinate and acetate when the TCA cycle becomes overflown, (3) initiation of biosynthesis with succinyl-CoA and acetyl-CoA, and (4) established succinic acid production. Here, we implemented the reverse β-oxidation pathway inC. glutamicumand assessed its functionality for adipic acid biosynthesis.ResultsTo obtain a non-decarboxylative condensation product of acetyl-CoA and succinyl-CoA, and to subsequently remove CoA from the condensation product, we introduced heterologous 3-oxoadipyl-CoA thiolase and acyl-CoA thioesterase intoC. glutamicum. No 3-oxoadipic acid could be detected in the cultivation broth, possibly due to its endogenous catabolism. To successfully biosynthesize and secrete 3-hydroxyadipic acid, 3-hydroxyadipyl-CoA dehydrogenase was introduced. Addition of 2,3-dehydroadipyl-CoA hydratase led to biosynthesis and excretion oftrans-2-hexenedioic acid. Finally, trans-2-enoyl-CoA reductase was inserted to yield 37 µg/L of adipic acid.ConclusionsIn the present study, we engineered the reverse β-oxidation pathway inC. glutamicumand assessed its potential for producing adipic acid from glucose as starting material. The presence of adipic acid, albeit small amount, in the cultivation broth indicated that the synthetic genes were expressed and functional. Moreover, 2,3-dehydroadipyl-CoA hydratase and β-ketoadipyl-CoA thiolase were determined as potential target for further improvement of the pathway.

허혈 재관류 손상은 기관 이식, 외과적 혈관 재개통 및 출혈 시에 발생하며 조직 및 기관 기능 장애를 유발한다. 최근 허혈 재관류 손상의 기전적 연구를 위해 간장 허혈 모델을 많이 이용하고 있다. 본 연구는 허혈 재관류 랫드 모델을 이용하여 항산화와 항염증 효과를 가진 것으로 알려진 Vanillin에 의한 간장 및 신장 손상에 대한 보호 효과를 알아보고 관련된 기전을 조사하기 위하여 실시하였다. 시험물질은 각각 100 mg/kg의 농도로 3일간 투여한 후, 60분동안 간을 결찰하여 허혈 재관류를 유도하여 관찰하였으며, 음성대조군, sham대조군 및 허혈재관류만 실시한 허혈 재관류대조군을 따로 두어, 약물투여군과 비교하였다. Vanillin 처치군에서는 AST, ALT 활성이 허혈 재관류대조군에 비해 유의하게 억제되었고, 조직병리학적 관찰에서도 염증 부분과 괴사부분이 현저하게 감소하였다. MDA와 SOD는 허혈 재관류군에 비해 유의적인 변화를 보였다. 이상의 결과를 종합하면 Vanillin은 간장 허혈 재관류에 의한 세포염증 및 세포괴사를 완화시켜 간세포 보호작용을 나타내었고, 신장의 사구체 및 원 위세뇨관에 염증 변화를 완화 시키고 있어 세포 손상을 방어하는 것으로 생각되며, 이러한 방어효과는 항산화 기능에 의한 영향으로 사료된다.

Extracts of several plants possess antithrombotic effects. Herein, we examined the antithrombotic effects of different extracts of Artemisia princeps Pampanini prepared using distilled water, hot distilled water, 70% ethanol, or subcritical water. The antithrombotic effects were determined using a co-culture system consisting of tumor necrosis factor-alpha (TNF-α)-treated EA.hy926 cells and THP-1 cells. In addition, the coagulation time of plasma collected from healthy volunteers was evaluated in terms of the prothrombin time and activated partial thromboplastin time. A carotid arterial thrombosis model was induced by ferric chloride in Sprague Dawley rats. The rats were treated with either sterile water or three different doses of the subcritical water extract for 2 weeks. The thrombus weight, gene expression of cell adhesion molecules, and histological characteristics were assessed. The results of in vitro studies revealed a significant inhibition in the adhesion of monocytes to EA.hy926 cells stimulated by TNF-α in the subcritical water extract-treated group. We also observed considerable suppression of the occlusion and mRNA expression of cell adhesion molecules in the in vivo experiments. This study suggests that Artemisia princeps Pampanini may have the potential to improve blood coagulation.

Background We describe the Hook technique enabling coil embolization in unfavorable M1 bifurcation aneurysms and analyze the morphologic variations in M1 bifurcation to evaluate how often such aneurysms can be seen. Methods Among 42 MCA aneurysms treated by stent-assisted coil embolization, aneurysms arising at the acute-angled hypoplastic M1 branch (n = 14) were treated by the Hook technique, in which a short stent was deployed only to protect the aneurysm neck after microcatheter selection of the hypoplastic M2, followed by subsequent coiling of the aneurysm. Outcome was evaluated, including Raymond classification, coil packing density, final modified Rankin Scale (mRS), and recurrence. Separately, 100 middle cerebral artery (MCA) bifurcation aneurysms were analyzed to assess the proportion of such unfavorable aneurysms. Results Procedural success of the Hook technique was obtained in 13 of 14 patients (93%). A mean packing density of 30% was achieved. Magnetic resonance angiographic follow-up at a median 4 months (range, 1–26 months) showed complete occlusion in 11 patients and residual neck filling in 3 patients. There was no clinical event (mRS = 0) over a median 17 months (range, 2–26 months) of clinical follow-up. One patient had a thrombotic occlusion during the procedure, which was resolved after tirofiban infusion, without evidence of an infarct or deficit. Of the 100 MCA bifurcation aneurysms, aneurysm arising in asymmetric hypoplastic M2 division was the most common type (48%). Conclusions The Hook technique enabled stent-assisted coiling of M1 bifurcation aneurysm with extension along the asymmetric hypoplastic M2 division and also securing the M2 branch.

Streptomyces griseus S4-7 is representative of strains responsible for the specific soil suppressiveness of Fusarium wilt of strawberry caused by Fusarium oxysporum f. sp. fragariae. Members of the genus Streptomyces secrete diverse secondary metabolites including lantipeptides, heat-stable lanthionine-containing compounds that can exhibit antibiotic activity. In this study, a class II lantipeptide provisionally named grisin, of previously unknown biological function, was shown to inhibit F. oxysporum. The inhibitory activity of grisin distinguishes it from other class II lantipeptides from Streptomyces spp. Results of quantitative reverse transcription-polymerase chain reaction with lanM-specific primers showed that the density of grisin-producing Streptomyces spp. in the rhizosphere of strawberry was positively correlated with the number of years of monoculture and a minimum of seven years was required for development of specific soil suppressiveness to Fusarium wilt disease. We suggest that lanM can be used as a diagnostic marker of whether a soil is conducive or suppressive to the disease.

In this study, we generated water and phosphate buffer treated with microwave plasma-generated gas in which the major component was nitric oxide (PGNO), and investigated the efficiency of the treated water and buffer in fertilization and sanitation. Real time NO level monitored by an electrode sensor was linearly increased over PGNO injection time, and removal of O2 from liquid before PGNO injection accelerated NO assimilation into liquids. Residual NO was still present 16 h after PGNO injection was stopped. H2O2, NO2−, and NO3− were also detected in PGNO-treated liquids. Spinach plants applied with 10 and 30 times diluted PGNO-treated water and 0.5 mM phosphate buffer showed slightly higher height and dry weight than control after 5 weeks. Plants grown with 10 and 30 times diluted PGNO-treated water exhibited the increased tolerance to water deficiency. Significant anti-microbial activity within 1 h was observed in un-diluted and in half-diluted PGNO-treated water and 0.5 mM phosphate buffer. Our results suggest that water or phosphate buffer containing NO, H2O2, NO2−, and NO3− can be produced by PGNO treatment, and that PGNO-treated water or buffer can be used as a potential fertilizer enhancing plant vitality with sanitation effect.

Adipic acid is a platform chemical, and is the most important commercial dicarboxylic acid. It has been targeted for biochemical conversion as an alternative to present chemical production routes. From the perspective of bioeconomy, several kinds of raw material are of interest including the sugar platform (derived from starch, cellulose or hemicellulose), the lignin platform (aromatics) and the fatty acid platform (lipid derived). Two main biochemical-based production schemes may be employed: (i) direct fermentation to adipic acid, or (ii) fermentation to muconic or glucaric acid, followed by chemical hydrogenation (indirect fermentation). This review presents a comprehensive description of the metabolic pathways that could be constructed and analyzes their respective theoretical yields and metabolic constraints. The experimental yields and titers obtained so far are low, with the exception of processes based on palm oil and glycerol, which have been reported to yield up to 50 g and 68 g adipic acid/L, respectively. The challenges that remain to be addressed in order to achieve industrially relevant production levels include solving redox constraints, and identifying and/or engineering enzymes for parts of the metabolic pathways that have yet to be metabolically demonstrated. This review provides new insights into ways in which metabolic pathways can be constructed to achieve efficient adipic acid production. The production host provides the chassis to be engineered via an appropriate metabolic pathway, and should also have properties suitable for the industrial production of adipic acid. An acidic process pH is attractive to reduce the cost of downstream processing. The production host should exhibit high tolerance to complex raw material streams and high adipic acid concentrations at acidic pH.

P-glycoprotein (P-gp) inhibition has been studied to overcome multidrug resistance in cancer chemotherapy but failed in clinical trials due to low/toxic effects. Recently, a dual modulation of transporters and natural derivatives have been examined to surmount this limitation. We examined breast cancer resistance protein (BCRP) inhibition in vitro and in vivo by P-gp inhibitors derived from natural compounds in previous studies. P-gp inhibitors increased the accumulation of the anticancer drug, topotecan (TPT)—a substrate of P-gp and BCRP, albeit with higher affinity for BCRP—in BCRP-overexpressing cells, resulting in cell death. These dual inhibitors, when orally co-administered with TPT, enhanced TPT bioavailability with slightly reduced total oral clearance (Clt/F) in rats. In xenograft mice, they strengthened oral TPT-induced tumor reduction with no alterations in body weight. Moreover, we investigated the effects of an oral drug formulation (Cremophor® EL, Tween® 80, and polyethylene glycol 400) on the transporters function. The excipients increased TPT accumulation in P-gp- or BCRP-overexpressing cells. Oral TPT bioavailability was higher with the formulation than with a control, as shown by the increases in the maximum plasma concentration (Cmax) and the area under the plasma concentration–time curve from zero to infinity (AUCINF) (p&lt, 0.01). Therefore, oral TPT bioavailability was enhanced by P-gp/BCRP dual inhibition, which resulted in a formulation-mediated increase in absorption and decrease in elimination, and a dual inhibitor-mediated decrease in elimination. These results suggest that the combination of dual inhibition by a natural derivative and the drug formulation can be a useful clinical approach.

Relationships between heat shock protein 27 (HSP27) and cancer aggressiveness, metastasis, drug resistance, and poor patient outcomes in various cancer types including non-small cell lung cancer (NSCLC) were reported, and inhibition of HSP27 expression is suggested to be a possible strategy for cancer therapy. Unlike HSP90 or HSP70, HSP27 does not have an ATP-binding pocket, and no effective HSP27 inhibitors have been identified. Previously, NSCLC cancer cells were sensitized to radiation and chemotherapy when co-treated with small molecule HSP27 functional inhibitors such as zerumbone (ZER), SW15, and J2 that can induce abnormal cross-linked HSP27 dimer. In this study, cancer inhibition effects of NA49, a chromenone compound with better solubility, longer circulation time, and less toxicity than J2, were examined in combination with anticancer drugs such as cisplatin and gefitinib in NSCLC cell lines. When the cytotoxic drug cisplatin was treated in combination with NA49 in epidermal growth factor receptors (EGFRs) WT cell lines, sensitization was induced in an HSP27 expression-dependent manner. With gefitinib treatment, NA49 showed increased combination effects in both EGFR WT and Mut cell lines, also with HSP27 expression-dependent patterns. Moreover, NA49 induced sensitization in EGFR Mut cells with a secondary mutation of T790M when combined with gefitinib. Augmented tumor growth inhibition was shown with the combination of cisplatin or gefitinib and NA49 in nude mouse xenograft models. These results suggest the combination of HSP27 inhibitor NA49 and anticancer agents as a candidate for overcoming HSP27-mediated drug resistance in NSCLC patients.

Convincing studies demonstrated that vaginal flora is one of the most impactful key components for the well-being of the genital tract in women. Nevertheless, the potential capability of vaginal-derived bacterial communities as biomarkers to monitor cervical carcinogenesis (CC) has yet to be studied actively compared to those of bacterial vaginosis (BV). We hypothesized that vaginal microbiota might be associated with the progression of CC. In this study, we enrolled 23 participants, including healthy controls (HC group, n = 7), patients with cervical intraepithelial neoplasia (CIN) 2 and 3 (CIN group, n = 8), and patients with invasive cervical cancer (CAN group, n = 8). Amplicon sequencing was performed using the Ion Torrent PGM to characterize the vaginal microbiota. Patients with CIN and CAN presented vaginal microbiota dysbiosis compared with HC. The alpha diversity analysis revealed that CC has a trend to be increased in terms of diversity indexes. Moreover, CC was associated with the abundance of specific microbes, of which Lactobacillus and Gardnerella were the most significantly different between HC and CIN, whereas Streptococcus was differentially abundant in CAN compared with CIN. We then evaluated their diagnostic abilities. Testing in terms of diagnostic ability using the three genera revealed considerably high performance with an area under the receiver-operating characteristic curve of 0.982, 0.953, and 0.922. The current study suggests that the presence of Gardnerella and Streptococcus may be involved in the advancment of CC.

The skin microbiome, especially the axillary microbiome, consists of odor-causing bacteria that decompose odorless sweat into malodor compounds, which contributes to the formation of body odor. Plant-derived products are a cheap source of bioactive compounds that are common ingredients in cosmetics. Microbial bioconversion of natural products is an ecofriendly and economical method for production of new or improved biologically active compounds. Therefore, in this study, we tested the potential of a Lactobacillus acidophilus KNU-02-mediated bioconverted product (BLC) of Lotus corniculatus seed to reduce axillary malodor and its effect on the associated axillary microbiota. A chemical profile analysis revealed that benzoic acid was the most abundant chemical compound in BLC, which increased following bioconversion. Moreover, BLC treatment was found to reduce the intensity of axillary malodor. We tested the axillary microbiome of 18 study participants, divided equally into BLC and placebo groups, and revealed through 16S rRNA gene sequencing that Staphylococcus, Corynebacterium, and Anaerococcus were the dominant taxa, and some of these taxa were significantly associated with axillary malodor. After one week of BLC treatment, the abundance of Corynebacterium and Anaerococcus, which are associated with well-known odor-related genes that produce volatile fatty acids, had significantly reduced. Likewise, the identified odor-related genes decreased after the application of BLC. BLC treatment enhanced the richness and network density of the axillary microbial community. The placebo group, on the other hand, showed no difference in the microbial richness, odor associated taxa, and predicted functional genes after a week. The results demonstrated that BLC has the potential to reduce the axillary malodor and the associated odor-causing bacteria, which makes BLC a viable deodorant material in cosmetic products.

Background Pseudomyopia is caused by increased refractive power by ciliary muscle spasm. Most patients cannot overcome pseudomyopia spontaneously; therefore, treatment of pseudomyopia is fastidious and needs a multidisciplinary approach. We report a case of unusual pseudomyopia with paradoxical accommodation, straining eyes to induce emmetropia at far distance and relaxing eyes to focus at near objects, contrary to physiological accommodation. Case presentation A 33-year-old woman experienced intermittent distant vision discomfort. This occurred at least a few hundred times daily. She could see near objects clearly; however, distant objects could be seen clearly only when she strained her eyes. Uncorrected distance visual acuity was 20/20 and manifest refraction (MR) in both eyes in the relaxed state was approximately − 2.5 D. MR changed to approximately − 0.5 D when she grimaced and strained her eyes when attempting to focus on distant letters. Her response was contrary to the physiological accommodative response. Cycloplegic refraction was approximately 0.0 D. Binocular autorefractor/keratometer was used to objectively evaluate her refractive response and pupil reaction according to accommodative stimulation. The IOL Master was used to evaluate the anterior chamber depth (ACD), lens thickness (LT), and pupil diameter with relaxed and strained eyes. For stepwise static accommodative stimuli (1–5 D), the refractive responses were correspondingly stepwise, similar to those elicited by healthy individuals. However, contrary to physiological accommodation, she strained her eyes to see distant objects and relaxed them to see near objects. There was no change in pupil diameter despite the accommodative stimuli being maximum. Biometry results showed that ACD deepened and LT flattened with eye strain, which were contrary to those during physiological accommodation. Conclusions We report a rare case with reverse of physiological accommodative response. When patients complain of unusual distant visual discomfort, pseudomyopia with paradoxical accommodation should be considered.

The fecal microbiota is being increasingly implicated in the diagnosis of various diseases. However, evidence on changes in the fecal microbiota in invasive cervical cancer (ICC) remains scarce. Here, we aimed to investigate the fecal microbiota of our cohorts, develop a diagnostic model for predicting early ICC, and identify potential fecal microbiota-derived biomarkers using amplicon sequencing data. We obtained fecal samples from 29 healthy women (HC) and 17 women with clinically confirmed early ICC (CAN). Although Shannon&rsquo, s diversity index was not reached at statistical significance, the Chao1 and Observed operational taxonomic units (OTUs) in fecal microbiota was significantly different between CAN and HC group. Furthermore, there were significant differences in the taxonomic profiles between HC and CAN, Prevotella was significantly more abundant in the CAN group and Clostridium in the HC group. Linear discriminant analysis effect size (LEfSe) analysis was applied to validate the taxonomic differences at the genus level. Furthermore, we identified a set of seven bacterial genera that were used to construct a machine learning (ML)-based classifier model to distinguish CAN from patients with HC. The model had high diagnostic utility (area under the curve [AUC] = 0.913) for predicting early ICC. Our study provides an initial step toward exploring the fecal microbiota and helps clinicians diagnose.

The purpose of the present study was to investigate the anti-inflammatory effects of a sugar isomer, leucrose (D-glucopyranosyl-α-(1–5)-D-fructopyranose) in a dextran sulfate sodium (DSS)-induced colitis mouse model and in RAW 264.7 macrophages. Colitis was induced in vivo with administration of two cycles of DSS (2.5%). Two groups of mice received an AIN-93G diet with 25% or 50% of the total sucrose content replaced with leucrose. This leucrose supplementation improved disease activity index (DAI) scores, colon length, histopathological damage, and myeloperoxidase (MPO) activity levels. In addition, the expression of pro-inflammatory mediators and cytokines decreased in the leucrose supplementation groups compared with the DSS alone treatment group. Furthermore, leucrose supplementation increased M2 macrophage polarization. Leucrose treatment suppressed IL-4-induced M2 polarization and increased JAK1/STAT6 phosphorylation in RAW264.7 cells. Taken together, these results indicate that leucrose exerts an anti-inflammatory effect by regulating M2 macrophage polarization via inflammatory cytokines and the JAK1/STAT6 signaling pathway.

Quorum sensing (QS) enables bacteria to organize gene expression programs, thereby coordinating collective behaviors. It involves the production, release, and population-wide detection of extracellular signaling molecules. The cellular processes regulated by QS in bacteria are diverse and may be used in mutualistic coordination or in response to changing environmental conditions. Here, we focused on the influence of the QS-dependent genes of our model bacterial strain Serratia fonticola GS2 on potential plant growth promoting (PGP) activities including indole-3-acetic acid (IAA) production, 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity, and biofilm formation. Based on genomic and phenotypic experimental data we identified and investigated the function of QS genes in the genome of the model strain. Our gene deletion study confirmed the biological functionality of the QS auto-inducer (gloI) and receptor (gloR) on potential PGP activities of GS2. A transcriptomic approach was also undertaken to understand the role of QS genes in regulation of genes primarily involved in PGP activities (IAA, ACC deaminase activity, and biofilm formation). Both transcriptomic and phenotypic data revealed that the QS-deletion mutants had considerably less PGP activities, as compared to the wild type. In addition, in vivo plant experiments showed that plants treated with GS2 had significantly higher growth rates than plants treated with the QS-deletion mutants. Overall, our results showed how QS-dependent genes regulate the potential PGP activities of GS2. This information may be helpful in understanding the relationship between QS-dependent genes and the PGP activity of bacteria, which aid in the production of practical bio-fertilizers for plant growth promotion.

We identified an antimicrobial peptide (AMP) from Lactobacillus acidophilus that was antagonistic to Aeromonas hydrophila. In vitro studies such as well-diffusion and field trials revealed that the AMP was active against A. hydrophila. The field trials of AMP using A. hydrophila-infected Channa striatus with a mannone oligosaccharide (MOS) prebiotic, A. hydrophila antigens, A. hydrophila-infected fish serum, L. acidophilus, and Lactobacillus cell free-supernatant (LABS-CFS) on an indicator organism further revealed that the antimicrobial agent could protect C. striatus. Other than the AMP, none of the above were able to eliminate the infectious agent A. hydrophila, and were only able to delay the death rate for 3–4 days. Thus, we conclude that the AMP is antagonistic to A. hydrophila and may be used for treatment of A. hydrophila infections. Subsequent L. acidophilus whole-genome sequence analyses enabled an understanding of the (probable) gene arrangement and its location on the chromosome. This information may be useful in the generation of recombinant peptides to produce larger quantities for treatment.

The intestinal tract is one of the most sensitive organs following irradiation. The protective effect of specific indigenous microbiota on irradiation-induced damage to intestinal epithelial cells has not been reported. Mice were irradiated with a single dose of 6 Gy of gamma rays. The intestinal damage was analyzed by histopathology. Intestinal stemness and differentiation were determined by intestinal organoid culture. Microbiota community was observed by high-throughput 16S rRNA gene sequencing and oligotyping analysis. We showed that distal small intestine was damaged by sublethal dose of gamma irradiation. Intestinal organoids derived from the irradiated mice showed defects in budding and mucin expression, suggesting the detrimental effect of irradiation on the intestinal stemness and differentiation. In addition, irradiation reduced intestinal immunoglobulin A level, concomitant with decreased microbiota diversity based on our high-throughput 16S rRNA gene sequencing data. Especially, the relative abundance of Lactobacillus was reduced at early time point post-irradiation; however, it was recovered at late time point. Oligotyping analysis within the Lactobacillus genus indicated that Lactobacillus-related oligotype 1 (OT1) including Lactobacillus acidophilus might drive recovery after irradiation as it was associated with increased long-term numbers post-exposure. We showed that treatment with heat-killed L. acidophilus rescued the budding-impaired organoids and induced sufficient differentiation in epithelial cells, and particularly mucin-producing cells, in intestinal organoids. This study provides the first evidence that the indigenous gut bacteria L. acidophilus enhance intestinal epithelial function with respect to irradiation-induced intestinal damage by improving intestinal stem cell function and cell differentiation.

Environmental factors can influence the composition of gut microbiota, but understanding the combined effect of lifestyle factors on adult gut microbiota is limited. Here, we investigated whether changes in the modifiable lifestyle factors, such as cigarette smoking, alcohol consumption, sleep duration, physical exercise, and body mass index affected the gut microbiota of Korean navy trainees. The navy trainees were instructed to stop smoking and alcohol consumption and follow a sleep schedule and physical exercise regime for eight weeks. For comparison, healthy Korean civilians, who had no significant change in lifestyles for eight weeks were included in this study. A total of 208 fecal samples were collected from navy trainees (n = 66) and civilians (n = 38) at baseline and week eight. Gut flora was assessed by sequencing the highly variable region of the 16S rRNA gene. The &alpha, and &beta, diversity of gut flora of both the test and control groups were not significantly changed after eight weeks. However, there was a significant difference among individuals. Smoking had a significant impact in altering &alpha, diversity. Our study showed that a healthy lifestyle, particularly cessation of smoking, even in short periods, can affect the gut microbiome by enhancing the abundance of beneficial taxa and reducing that of harmful taxa.

Background Multifocal spherical nonstaining cavities and gram-positive, rod-shaped, and endospore-forming bacteria were found in the liver of a sow that died suddenly. Clostridium novyi type B was identified and isolated from the sudden death case, and the isolate was characterized by molecular analyses and bioassays in the current study. Results C. novyi was isolated from the liver of a sow that died suddenly and was confirmed as C. novyi type B by differential PCR. The C. novyi isolate fermented glucose and maltose and demonstrated lecithinase activity, and the cell-free culture supernatant of the C. novyi isolate exhibited cytotoxicity toward Vero cells, demonstrating that the isolate produces toxins. In addition, whole-genome sequencing of the C. novyi isolate was performed, and the complete sequences of the chromosome (2.29 Mbp) and two plasmids (134 and 68 kbp) were identified for the first time. Based on genome annotation, 7 genes were identified as glycosyltransferases, which are known as alpha toxins; 23 genes were found to be related to sporulation; 12 genes were found to be related to germination; and 20 genes were found to be related to chemotaxis. Conclusion C. novyi type B was isolated from a sow in a sudden death case and confirmed by biochemical and molecular characterization. Various virulence-associated genes were identified for the first time based on whole-genome sequencing.

4-Amino-1-butanol (4AB) serves as an important intermediate compound for drugs and a precursor of biodegradable polymers used for gene delivery. Here, we report for the first time the fermentative production of 4AB from glucose by metabolically engineered Corynebacterium glutamicum harboring a newly designed pathway comprising a putrescine aminotransferase (encoded by ygjG) and an aldehyde dehydrogenase (encoded by yqhD) from Escherichia coli, which convert putrescine to 4AB. Application of several metabolic engineering strategies such as fine-tuning the expression levels of ygjG and yqhD, eliminating competing pathways, and optimizing culture condition further improved 4AB production. Fed-batch culture of the final metabolically engineered C. glutamicum strain produced 24.7 g/L of 4AB. The strategies reported here should be useful for the microbial production of primary amino alcohols from renewable resources. This article is protected by copyright. All rights reserved.

A microbial imbalance called dysbiosis leads to inflammatory bowel disease (IBD), which can include ulcerative colitis (UC). Fecal microbiota transplantation (FMT), a novel therapy, has recently been successful in treating gut dysbiosis in UC patients. For the FMT technique to be successful, the gut microbiota of both the healthy donors and UC patients must be characterized. For decades, next-generation sequencing (NGS) has been used to analyze gut microbiota. Despite the popularity of NGS, the cost and time constraints make it difficult to use in emergency services and activities related to the periodic monitoring of microbiota profile alterations. Hence, in this study, we developed a multiplex TaqMan qPCR assay (MTq-PCR) with novel probes to simultaneously determine the relative proportions of the three dominant microbial phyla in the human gut: Bacteroidetes, Firmicutes, and Proteobacteria. The relative proportions of the three phyla in fecal samples of either healthy volunteers or UC patients were similar when assessed NGS and the MTq-PCR. Thus, our MTq-PCR assay could be a practical microbiota profiling alternative for diagnosing and monitoring gut dysbiosis in UC patients during emergency situations, and it could have a role in screening stool from potential FMT donors.

The enzymatic reactions leading to the deamination of β-lysine, lysine, or 2- aminoadipic acid are of great interest for the metabolic conversion of lysine to adipic acid. Enzymes able to carry out these reactions are not known, however ammonia lyases (EC 4.3.1.-) perform deamination on a wide range of substrates. We have studied 3-methylaspartate ammonia lyase (MAL, EC 4.3.1.2) as a potential candidate for protein engineering to enable deamination towards β-lysine, that we have shown to be a competitive inhibitor of MAL. We have characterized MAL activity, binding and inhibition properties on six different compounds that would allow to define the molecular determinants necessary for MAL to deaminate our substrate of interest. Docking calculations showed that β-lysine as well as the other compounds investigated could fit spatially into MAL catalytic pocket, although they probably are weak or very transient binders and we identified molecular determinants involved in the binding of the substrate. The hydrophobic interactions formed by the methyl group of 3-methylaspartic acid, together with the presence of the amino group on carbon 2, play an essential role in the appropriate binding of the substrate. The results showed that β-lysine is able to fit and bind in MAL catalytic pocket and can be potentially converted from inhibitor to substrate of MAL upon enzyme engineering. The characterization of the binding and inhibition properties of the substrates tested here provide the foundation for future and more extensive studies on engineering MAL that could lead to a MAL variant able to catalyse this challenging deamination reaction.

The velvet regulator VosA plays a pivotal role in asexual sporulation in the model filamentous fungus Aspergillus nidulans. In the present study, we characterize the roles of VosA in sexual spores (ascospores) in A. nidulans. During ascospore maturation, the deletion of vosA causes a rapid decrease in spore viability. The absence of vosA also results in a lack of trehalose biogenesis and decreased tolerance of ascospores to thermal and oxidative stresses. RNA-seq-based genome-wide expression analysis demonstrated that the loss of vosA leads to elevated expression of sterigmatocystin (ST) biosynthetic genes and a slight increase in ST production in ascospores. Moreover, the deletion of vosA causes upregulation of additional gene clusters associated with the biosynthesis of other secondary metabolites, including asperthecin, microperfuranone, and monodictyphenone. On the other hand, the lack of vosA results in the downregulation of various genes involved in primary metabolism. In addition, vosA deletion alters mRNA levels of genes associated with the cell wall integrity and trehalose biosynthesis. Overall, these results demonstrate that the velvet regulator VosA plays a key role in the maturation and the cellular and metabolic integrity of sexual spores in A. nidulans.

Many bacteria found in the rhizosphere provide contribution for the host plant’s growth and protection that are known as plant growth-promoting rhizobacteria (PGPR). Plant–microbe interactions in the rhizosphere are important factors in determining the health of plants. Research for commercialization of these PGPR as an alternative to the use of chemical fertilizers for a more environmentally friendly treatment is continuously being improved. In this review, we discuss the essential traits that rhizobacteria must possess for them to be considered PGPR and report the bacterial species that exhibit these essential plant growth-promoting activities and which are approved for use by the South Korean regulations.

The prevalence of inflammatory bowel disease (IBD) is increasing worldwide, and the key risk factor is high sugar diet. The objective of this study was to investigate anti-inflammatory effects of turanose on IBD in a dextran sulfate sodium (DSS)-induced colitis mouse model. An amylosucrase from Neisseria sicca (NsiAS) was newly identified and successfully expressed. This enzyme exhibited optimal activity at 45 °C and pH 8.5. By utilizing this recombinant enzyme expressed in the E. coli host, turanose yield maximally reached 47% from 2.0 M sucrose. Colitis was induced with two 5-d cycles of 2.5% DSS in drinking water with a 10-d inter-cycle interval. Mice were fed AIN93G diet with 25% and 50% turanose replacement. Improved disease activity index scores, colon length, histopathological features, and myeloperoxidase activity were observed in the turanose supplemented group. Meanwhile, decreases in micro RNA (miR)-21 expression, histone acetylation, expression of pro-inflammatory cytokines, and phosphorylated ERK and STAT3 were observed in colon tissues. Notably, miR-21 and histone acetylation levels were found to positively correlated with expression of tumor necrosis factor α. In conclusion, turanose can attenuate colitis via regulatory effects on miRNA-21 expression and histone acetylation-related proinflammatory mediators. These findings also support further exploration of turanose as a potential therapeutic sugar substitute for patients with IBD.

An overdose of acetaminophen (AAP) causes hepatic and renal toxicity. This study examined the protective effects of Gastrodia elata Blume (GEB) on hepatic and renal injury induced by AAP. Rats were orally administered distilled water or GEB for 14 days and injected with AAP 1 h after the oral last administration; control rats were administered water without AAP injection. All rats were sacrificed 24 h after AAP injection. The GEB pretreatment group showed decreased necrosis and the expression of pro-inflammatory cytokines in the liver and kidney. TUNEL-positive cells and oxidative stress marker, such as malondialdehyde, were decreased. However, antioxidant enzymes, such as glutathione and superoxide dismutase, were increased. The expression of CYP2E1 and N-acetyl-beta-d-glucosaminidase was decreased in the GEB pretreatment group. This study shows that GEB prevents AAP-induced liver and kidney injury.

The enzyme xylose isomerase (E.C. 5.3.1.5, XI) is responsible for the conversion of an aldose to ketose, especially xylose to xylulose. Owing to the ability of XI to isomerize glucose to fructose, this enzyme is used in the food industry to prepare high-fructose corn syrup. Therefore, we studied the characteristics of XI from Anoxybacillus kamchatkensis G10, a thermophilic bacterium. First, the gene coding for XI (xylA) was inserted into the pET-21a(+) expression vector and the construct was transformed into the Escherichia coli competent cell BL21 (DE3). The expression of recombinant XI was induced in the absence of isopropyl-thio-β-galactopyranoside and purified using Ni-NTA affinity chromatography. The optimum temperature of recombinant XI was 80°C and measurement of the heat stability indicated that 55% of residual activity was maintained after 2 h incubation at 60°C. The optimum pH was found to be 7.5 in sodium phosphate buffer. Magnesium, manganese, and cobalt ions were found to increase the enzyme activity; manganese was the most effective. Additionally, recombinant XI was resistant to the presence of Ca²⁺ and Zn²⁺ ions. The kinetic properties, Km and Vmax, were calculated as 81.44 mM and 2.237 μmol/min/mg, respectively. Through redundancy analysis, XI of A. kamchatkensis G10 was classified into a family containing type II XIs produced by the genera Geobacillus, Bacillus, and Thermotoga. These results suggested that the thermostable nature of XI of A. kamchatkensis G10 may be advantageous in industrial applications and food processing.

Phyllodulcin, an isocoumarin derivatives and well-known natural sweetener derived from Hydrangea macrophylla var, was evaluated for its roles in diabetes-related metabolic and genetic changes by regulating glucose homeostasis in the liver. C57BL/KsJ-db/db mice received 20 mg/kg body weight (b.w.) stevioside (SVS 20), 10 mg/kg b.w. phyllodulcin (P 10), or 20 mg/kg b.w. phyllodulcin (P 20) for four weeks. Water and food intake were significantly lower in the phyllodulcin supplemented group compared with the diabetic group. Phyllodulcin supplementation suppressed the levels of fasting blood glucose, HbA1c, and plasma and hepatic triglyceride level. Phyllodulcin improved hepatic lipogenesis, while also suppressing inflammation and oxidative stress. Phyllodulcin inhibited hepatic fibrosis and regulated liver glucose homeostasis. Thus, these results indicate the potential of phyllodulcin to serve as a therapeutic agent to improve global hepatic function and metabolic abnormalities of diabetic conditions and the necessity to be better explored.

Although the confirmation of inflammatory changes within tissues at the onset of various diseases is critical for the early detection of disease and selection of appropriate treatment, most therapies are based on complex and time-consuming diagnostic procedures. Raman spectroscopy has the ability to provide non-invasive, real-time, chemical bonding analysis through the inelastic scattering of photons. In this study, we evaluate the feasibility of Raman spectroscopy as a new, easy, fast, and accurate diagnostic method to support diagnostic decisions. The molecular changes in carrageenan-induced acute inflammation rat tissues were assessed by Raman spectroscopy. Volumes of 0 (control), 100, 150, and 200 µL of 1% carrageenan were administered to rat hind paws to control the degree of inflammation. The prominent peaks at [1,062, 1,131] cm-1 and [2,847, 2,881] cm-1 were selected as characteristic measurements corresponding to the C-C stretching vibrational modes and the symmetric and asymmetric C-H (CH2 ) stretching vibrational modes, respectively. Principal component analysis of the inflammatory Raman spectra enabled graphical representation of the degree of inflammation through principal component loading profiles of inflammatory tissues on a two-dimensional plot. Therefore, Raman spectroscopy with multivariate statistical analysis represents a promising method for detecting biomolecular responses based on different types of inflammatory tissues.

Burkholderia species are widely distributed across wide ecological niches. Many genera of Burkholderia are known to be associated with plants and are involved in processes such as suppression of soil-borne pathogens, acceleration of plant growth and endophytic colonization. In the present study, a strain belonging to the Burkholderia cepacia complex, which was termed JBK9, was isolated. The strain JBK9 showed broad-spectrum antifungal activities against Phytophthora capsici, Fusarium oxysporum, and Rhizoctonia solani, which are representative phytopathogenic fungi, inhibiting their growth by 59.56, 51.92, and 34.22%, respectively. The strain produced an antifungal compound that was confirmed to be pyrrolnitrin by TLC, HPLC, and NMR analyses. Using an in vitro assay for plant root colonization, we observed that the population densities of B. cepacia JBK9 on the upper 1 cm of host plant roots were significantly different between Burkholderia species. The high motility of these strains is likely to have contributed to their efficient root colonization. The isolated strain was evaluated in vivo for its ability to control Phytophthora blight via a pot test. Compared with Burkholderia strains KCTC 2973 and ATCC 25416, B. cepacia JBK9 demonstrated a stronger antifungal activity against P. capsici. The strain B. cepacia JBK9 could be further developed as a biological control agent for pepper plants.

Paenibacillus donghaensis JH8T (KCTC 13049T=LMG 23780T) is a Gram-positive, mesophilic, endospore-forming bacterium isolated from East Sea sediment at depth of 500m in Korea. The strain exhibited plant cell wall hydrolytic and plant growth promoting abilities. The complete genome of P. donghaensis strain JH8T contains 7602 protein-coding sequences and an average GC content of 49.7% in its chromosome (8.54Mbp). Genes encoding proteins related to the degradation of plant cell wall, nitrogen-fixation, phosphate solubilization, and synthesis of siderophore were existed in the P. donghaensis strain JH8T genome, indicating that this strain can be used as an eco-friendly microbial agent for increasing agricultural productivity.

Plant growth-promoting rhizobacteria (PGPR) affect plant growth through various mechanisms, such as indole-3-acetic acid (IAA) production, 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase activity, and biofilm formation. The aim of the study reported here was to isolate and characterize rhizobacteria that produce quorum-sensing signal molecules and other PGPR-related molecules. A biofilm-forming bacterium, GS2, was isolated from the rhizosphere of a sesame plant and subsequently found to produce two quorum-sensing signal molecules that were identified as N-hexanoyl-L-homoserine lactone (m/z 200) and N-octanoyl-L-homoserine lactone (m/z 228) by liquid chromatography–tandem mass spectrometry analysis. The strain was also found to produce IAA (17.2 μg mL−1), gibberellins (113.7 μg mL−1), and ACC deaminase (9.7 μM α-ketobutyrate mg−1 protein h−1). The strain was identified as Serratia glossinae based on a comparison of 16S rRNA gene sequences. Inoculation of the strain promoted growth of a gibberellin-deficient rice dwarf mutant (Waito-C). Different growth attributes, including shoot and root elongation, chlorophyll content, and plant weight could be attributed to the PGPR characteristics of strain GS2. These results suggest that S. glossinae strain GS2 can serve as a microbial agent that improves plant growth.

Abnormalities in the hyperbolic relationship between insulin sensitivity and insulin secretion may cause oxidative stress and non-enzymatic glycation, resulting in an increased risk of type 2 diabetes. Here, we performed a 14-week study to investigate the effects of ethanolic extract of Mori ramulus (MRE; 0, 800, and 1600 mg/kg body weight) and its signature component oxyresveratrol (OXY; 800 mg/kg body weight) on β-cell dysfunction and insulin resistance in C57BLKS/J db/db mice fed with a high-fat diet. Compared with the diabetic control group, the high-dose MRE group showed a significant decrease in fasting blood glucose ( p = 0.0024); a significant increase in insulin secretion as measured by insulin ( p = 0.0012) and C-peptide ( p = 0.0103) levels in plasma and insulin content ( p = 0.0440) and homeobox factor-1 protein expression ( p = 0.0148) in the pancreas; and a significant increase in insulin sensitivity as measured by insulin receptor mRNA expression in the liver ( p = 0.0179) and adipose tissue ( p = 0.0491). In addition, improvements in the reactive oxygen species level and inflammatory pancreatic and hepatic tissue damage were also observed in the MRE group as assessed by histological findings. A similar but weaker effect was found in the OXY group. Furthermore, we observed a potentiating effect of MRE and OXY on insulin secretion in INS-1 cells in the presence of 27 mM glucose, together with an anti-glycation effect as indicated by methylglyoxal-trapping capacity and inhibition of advanced glycation end-product formation. Taken together, these data suggest that MRE could ameliorate β-cell dysfunction and insulin resistance by reducing oxidative damage and advanced glycation end-product (Wagenknecht et al., 2003) formation and that these effects are due, at least in part, to OXY.

Denitrification is a crucial process for the global nitrogen cycle through the reduction of nitrates by heterotrophic bacteria. Denitrifying microorganisms play an important role in eliminating fixed nitrogen pollutants from the ecosystem, concomitant with N2O emission. Although many microbial denitrifiers have been identified, little is known about the denitrifying ability of the genus Geobacillus. Here, we report the first complete genome sequences of Geobacillus subterraneus KCTC 3922T, isolated from Liaohe oil field in China, and G. thermodenitrificans KCTC 3902T. The strain KCTC 3922T contains a complete set of genes involved in denitrification, cofactor biogenesis, and transport systems, which is consistent with a denitrifying activity. On the other hand, G. thermodenitrificans KCTC 3902T exhibited no denitrifying activity probably due to the lack of molybdnumtransferase (moeA) and nitrite transporter (nirC) genes. Therefore, comparative genome analysis of Geobacillus strains highlights the potential impact on treatment of nitrate-contaminated environments.

In this study, we investigated the cardioprotective effects of fermented garlic extract (FGE) against ischemia-reperfusion (IR) injury in H9c2 cells and myocardium. FGE used in this study is rich in nitrite (1.42 mg/ml of FGE) and reduced glutathione (GSH, 3.23 mg/ml of FGE). We compared the effectiveness of FGE against HR–induced oxidative stress with that of nitrite, GSH, and a mixture of nitrite and GSH (nitrite + GSH). In addition, we utilized fluorinated xerogel-derived oxygen microsensors to measure the changes in oxygen level (pO2) of the myocardium during myocardial IR by FGE. As a result, the pO2 in the FGE group restored to pre-ischemic level (75.1 ± 36.9%) at reperfusion period; however, that of the control group did not increase throughout reperfusion period (2.0 ± 1.7%, p

Current investigation conducted to evaluate the associated fungal endophyte interactions of a Cd hyper-accumulator Solanum nigrum Korean ecotype under varying concentrations of Cd. Two indole-3-acetic acid (IAA) producing fungal strains, RSF-4L and RSF-6L, isolated from the leaves of S. nigrum, were initially screened for Cd tolerance and accumulation potential. In terms of dry biomass production, the strain RSF-6L showed higher tolerance and accumulation capacity for Cd toxicity in comparison to RSF-4L. Therefore, RSF-6L was applied in vivo to S. nigrum and grown for six weeks under Cd concentrations of 0, 10, and 30mgKg-1 of dry sand. The effect of fungal inoculation assessed by plant physiological responses, endogenous biochemical regulations, and Cd profile in different tissues. Significant increase were observed in plant growth attributes such as shoot length, root length, dry biomass, leaf area, and chlorophyll contents in inoculated RSF-6L plants in comparison to non-inoculated plants with or without Cd contamination. RSF-6L inoculation decreased uptake of Cd in roots and above ground parts, as evidenced by a low bio-concentration factor (BCF) and improved tolerance index (TI). However, Cd concentration in the leaves remained the same for inoculated and non-inoculated plants under Cd spiking. Fungal inoculation protected the host plants, as evidenced by low peroxidase (POD) and polyphenol peroxidase (PPO) activities and high catalase (CAT) activity. Application of appropriate fungal inoculation that can improve tolerance mechanisms of hyper-accumulators and reduce Cd uptake can be recommended for phyto-stabilisation/immobilisation of heavy metals in crop fields.

The Silurus microdorsalis is known as Korean endemic slender catfish. Despite its value as a biological resource, there is no complete mitochondrial genome sequence. The complete mitochondrial geno...

In the anaerobic process, fat-oil-grease (FOG) is hydrolysed to long-chain fatty acids (LCFAs) and glycerol (GLYC), which are then used as substrates to produce biogas. The increase in FOG and LCFAs inhibits methanogenesis, and so far, most work investigating this inhibition has been carried out when FOG or LCFAs were used as co-substrates. In the current work, the inhibition of methanogenesis by FOG, LCFAs and GLYC was investigated when used as sole substrates. To gain more insight on the dynamics of this process, the change of microbial community was analysed using 16S rRNA gene amplicon sequencing. The results indicate that, as the concentrations of cooking olive oil (CO, which represents FOG) and LCFAs increase, methanogenesis is inhibited. For instance, at 0.01 g. L-1 of FOG, the rate of biogas formation was around 8 ml.L-1.day-1, and this decreased to = 45°C and NaCl > 3% led to a significant decrease in the rate of biogas formation. Microbial community analyses were carried out from samples from 3 different bioreactors (CO, OLEI and GLYC), on day 1, 5 and 15. In each bioreactor, microbial communities were dominated by Proteobacteria, Firmicutes and Bacteroidetes phyla. The most important families were Enterobacteriaceae, Pseudomonadaceae and Shewanellaceae (Proteobacteria phylum), Clostridiacea and Ruminococcaceae (Firmicutes) and Porphyromonadaceae and Bacteroidaceae (Bacteroidetes). In CO bioreactor, Proteobacteria bacteria decreased over time, while those of OLEI and GLYC bioreactors increased. A more pronounced increase in Bacteroidetes and Firmicutes were observed in CO bioreactor. The methanogenic archaea Methanobacteriaceae and Methanocorpusculaceae were identified. This analysis has shown that a set of microbial population is selected as a function of the substrate.

The main purpose of this study was to explore the potential influences of pickleweed vegetation on the abundance, diversity and metabolic activities of microbial communities in four distinct areas of a petroleum-contaminated solid waste management unit (SWMU) located in Contra Costa County, northern California. The four areas sampled include two central areas, one of which is central vegetated (CV) and one unvegetated (UV), and two peripheral vegetated areas, one of which is located to the west side of the SWMU (V-West) and one located to the east side (V-East). Measurements were made of total petroleum hydrocarbons (TPH), polyaromatic hydrocarbons (PAH), soil physicochemical properties, and various aspects of microbial communities including metabolic activities, microbial abundances (PLFAs), diversity and composition based on amplicon sequencing. The peripheral V-East and V-West sites had 10-times lower electrical conductivity (EC) than that of the CV and UV sites. The high salinity levels of the CV and UV sites were associated with significant reductions in bacterial and fungal abundances (PLFA) when compared to V-East but not when compared to V-West. TPH levels of CV and UV were not significantly different from those of V-West but were substantially lower than V-East TPH (19,311 mg/kg of dry soil), the high value of which may have been associated with a pipeline that ran through the area. Microbial activities (in terms of soil respiration and the activities of three soil enzymes, i.e., urease, lipase, and phosphatase) were greatest in the vegetated sites compared to the UV site. The prokaryotic community was not diverse as revealed by the Shannon index with no significant variation among the four groups of samples. However, the fungal community of the peripheral sites, V-East and V-West had significantly higher OTU richness and Shannon index. Structure of prokaryotic communities inhabiting the rhizosphere of pickleweed plants at the three sites differed significantly and were also different from those found in the UV region of the central site according to pairwise, global PERMANOVA and ANOSIM analyses. The differences in OTU-based rhizosphere-associated bacterial and fungal communities' composition were explained mainly by the changes in soil EC and pH. The results suggest that saline TPH-contaminated areas that are vegetated with pickleweed are likely to have increased abundances, diversity and metabolic activities in the rhizosphere compared to unvegetated areas, even in the presence of high salinity.

Background/Aims: Stress is closely related to the deterioration of digestive disease. Melatonin has potent anti-inflammatory properties. The objective of this study was to determine the effect of water stress (WS) and sleep deprivation (SD) on intestinal microbiota and roles of melatonin in stressful condition. Methods: We used C57BL/6 mice and specially designed water bath for stress and SD for 10 days. We measured melatonin concentrations in serum, feces, and colon tissues by high-performance liquid chromatography. Genomic DNA was extracted from feces and amplified using primers targeting V3 to V4 regions of bacterial 16S ribosomal RNA genes. Results: Compared to the control, melatonin concentration was lower in the WS and SD. Fecal concentration was 0.132 pg/mL in control, 0.062 pg/mL in WS, and 0.068 pg/mL in SD. In colon tissue, it was 0.45 pg/mL in control, 0.007 pg/mL in WS, and 0.03 pg/mL in SD. After melatonin treatment, melatonin concentrations in feces and colon tissue were recovered to the level of control. Metagenomic analysis of microbiota showed abundance in colitogenic microbiota in WS and SD. Melatonin injection attenuated this harmful effect. WS and SD showed decreased Lactobacillales and increased Erysipelotrichales and Enterobacteriales. Melatonin treatment increased Akkermansia muciniphila and Lactobacillus and decreased Bacteroides massiliensis and Erysipelotrichaceae. Conclusions: This study showed that stress and SD could affect intestinal dysbiosis and increase colitogenic microbiota, which could contribute to the aggravating digestive disease. Melatonin concentrations in feces and colon tissue decreased under WS and SD. Melatonin treatment brought recovery of melatonin concentration in colon tissue and modulating dysbiosis of intestinal microbiota.

The food industry produces highly perishable products. Food spoilage represents a severe problem for food manufacturers. Therefore, it is important to identify effective preservation solutions to prevent food spoilage. Ocins (e.g., bacteriocins, lactocins, and enterocins) are antibacterial proteins synthesized by bacteria that destroy or suppress the growth of related or unrelated bacterial strains. Ocins represent a promising strategy for food preservation, because of their antagonist effects toward food spoilage microorganisms, high potency, and low toxicity. Additionally, they can be bioengineered. The most common and commercially available ocins are nisin, plantaracin, sakacin P, and pediocin. Several ocins have been characterized and studied biochemically and genetically; however, their structure-function relationship, biosynthesis, and mechanism of action are not understood. This narrative review focuses primarily on ocins and their relevance to the food industry to help prevent food spoilage. In particular, the applications and limitations of ocins in the food industry are highlighted.

Traditional fermented alcoholic beverages are drinks produced locally using indigenous knowledge, and consumed near the vicinity of production. In Ethiopia, preparation and consumption of cereal- and fruit-based traditional fermented alcoholic beverages is very common. Tella, Borde, Shamita, Korefe, Cheka, Tej, Ogol, Booka, and Keribo are among the popular alcoholic beverages in the country. These beverages have equal market share with commercially produced alcoholic beverages. Fermentation of Ethiopian alcoholic beverages is spontaneous, natural and uncontrolled. Consequently, achieving consistent quality in the final product is the major challenge. Yeasts and lactic acid bacteria are the predominate microorganisms encountered during the fermentation of these traditional alcoholic beverages. In this paper, we undertake a review in order to elucidate the physicochemical properties, indigenous processing methods, nutritional values, functional properties, fermenting microorganisms and fermentation microbial dynamics of Ethiopian traditional alcoholic beverages. Further research will be needed in order to move these traditional beverages into large-scale production.

In this study, we investigate the biochemical characteristics of oxidative stress in age-related macular degeneration (AMD) and diabetic retinopathy (DR) by analyzing aqueous humors. Nondiabetic cataract aqueous humor was used as the control. The level of oxidative damage was evaluated based on changes in Raman spectral intensity. Seven prominent peaks were detected at 1002, 1043, 1062, 1352, 1419, 1454, and 1656 cm−1. We proposed four multimodal biomarkers to distinguish these peaks based on the ratios of Raman intensities in two wavelengths, including CHO (C–O stretching or C–O–H bending modes), AG (adenine and guanine), PRO-AG (protein and AG), and PHEα (phenylalanine symmetric ring breath and amide I α-helix) markers. The presence of oxidative damage was detected by CHO and AG markers associated with C–O stretching, C–O–H bending modes in carbohydrates (1043 cm−1), and the nucleic acids adenine and guanine (1352 cm−1), respectively. DR-related oxidative damage was identified by PRO-AG and PHEα markers associated with adenine, guanine, and protein components (1419 and 1454 cm−1) and amide I α-helix protein structure (1656 cm−1), respectively. AMD-related oxidative damage was identified by four biomarkers. Four multimodal biomarkers with simple linear threshold values achieved high sensitivity of 100% and high specificity of 100% for classifying oxidative stress-induced AMD and DR diseases. Therefore, Raman-based label-free optical detection is effective for detecting the presence of age-related or diabetic oxidative damage in aqueous humor.

선형 사상충의 일종인 심장사상충은 개의 심폐 사상충증을 유발한다. 이에 본 연구의 목적은 심장사상충을 효과적으로 검출할 수 있는 실시간 중합효소연쇄반응 기법을 개발함에 있다. 연구에 있어서 사용된 프라이머 및 프로브는 선행연구에서 제작된 심장사상충 특이 프라이머 및 새롭게 제작된 TaqMan 프로브를 이용하였다. 선행연구에서 제작된 프라이머 및 농도별로 희석된 게놈유전자와 플라스미드유전자가 SYBR Green 실시간 중합효소연쇄반응 수행에 이용되었으며, 중합효소연쇄반응 과정 중 증폭 이후의 녹는 곡선의 결과를 분석하였다. 분석결과 사용된 프라이머는 각각 게놈유전자 및 플라스미드 유전자에서 특이 녹는 곡선을 나타냄에 따라 심장사상충 특이 사이토크롬 C 산화효소 유전자만을 증폭하고 있음을 확인 할 수 있었다. 새롭게 제작된 TaqMan 프로브는 SYBR Green 실시간 중합효소연쇄반응과의 결과를 농도별로 희석된 플라스미드유전자를 이용하여 비교 분석하였고, 분석결과 TaqMan 프로브를 이용한 실시간 중합효소연쇄반응이 검출효율 및 특이도에 있어서 우수함을 확인할 수 있었다. 본 연구를 통하여 개발한 실시간 중합효소연쇄반응은 기존의 전통적인 진단기법의 한계를 극복할 수 있는 신속하고 정확한 향상된 진단기법을 제시한다.

Given the high prevalence of atherothrombotic disease and increasing attention to functional foods, we tested the hypothesis that a combination supplement (PS) containing Phellinus baumii (PB) and Salvia miltiorrhiza Bunge (SM) may improve the atherothrombotic profile in rats. The PS (Experiment 1) or PB/SM (Experiment 2) supplement was administered orally once daily for 7 days prior to inducing platelet activation at the last day. Orally administered PS showed an additive and positive effect on nitric oxide (NO) availability, adhesion molecule/pro-inflammatory cytokine release, and thrombosis compared to controls. This effect was attributed more to PB than to SM for cyclooxygenase (COX)-1 and to SM for COX-2. In vitro data revealed that tanshinone IIA derived from SM remarkably suppressed COX-2, and protocatechuic acid derived from PB caused a small increase in NO production. The data suggest the potential use of PS to ameliorate endothelial and vascular dysfunction induced by unintended platelet activation.