Bacterial diversity of baechu-kimchi with seafood based on culture-independent investigations.

Baechu-kimchi is a traditional Korean dish of fermented vegetables, in which kimchi cabbage is the major ingredient. Seafood is added to baechu-kimchi in coastal areas, giving this dish regional diversity. However, little is known about how the addition of seafood affects the bacterial diversity of kimchi. Therefore, in this study, the bacterial diversity of five varieties of baechu-kimchi with seafood and one variety of baechu-kimchi without seafood was analyzed using culture-independent techniques. In 81.7% of all kimchi analyzed, the predominant species were members of the phylum Firmicutes and the lactic acid bacteria, Latilactobacillus sakei, Leuconostoc mesenteroides, Pediococcus inopinatus, and Weissella koreensis. These organisms were similar to those identified in baechu-kimchi without the addition of seafood, which was used as a control group, and bacterial community of previously reported kimchi. Therefore, the results of this study confirmed that the addition of seafood did not significantly affect the bacterial community in baechu-kimchi.

The addition of jogi, Micropogonias undulates, affects amino acid content in kimchi fermentation.

The effects of jogi (the fish Atlantic croaker, Micropogonias undulatus) on the production of physicochemical components, such as color, organic acids, and amino acids, in kimchi, a traditional fermented vegetable food of Korea, were determined. As fermentation progressed, the color change of jogi-added kimchi increased, but in comparison with that of the control group without jogi-added kimchi, was difficult to distinguish with the naked eye. Reducing sugar decreased in all experimental groups, and as fermentation progressed, kimchi with jogi showed a lower value. Acetic acid, citric acid, lactic acid, and ethanol, were highly produced in both types of kimchi, and above all, the jogi-baechu-kimchi group showed higher acetic acid and lactic acid contents than the control group. The increase and decrease of amino acids were similar in both types of kimchi. However, significantly, immediately after manufacture, the savory components aspartic acid and glutamic acid were detected higher than the control group. Subsequently, the fermentation tended to decrease as it progressed, but the content was higher than that of the control group. The above results show that jogi addition has a greater effect on the contents of amino acid, especially the savory component, than on the physicochemical components.

Current status of the novel food ingredient safety evaluation system.

Increasing demand for new foods, technological development, and vegan market growth have led to an increase in new food ingredients, so the need for safety assessment of these ingredients is important. Representative safety assessment systems are the Generally Recognized as Safe (GRAS) notification of the Food and Drug Administration in the USA and the novel food system of the European Food Safety Authority in the European Union. GRAS is a notification system for information on food ingredients, food additives and functional foods under the responsibility of the applicant, while the novel food system assesses the safety of food ingredients excluding food additives. In Korea, a safety evaluation system is established for temporary food ingredients, which includes food ingredients without a domestic intake history. However, safety assessment systems for novel foods from other countries and food ingredients produced by the application of new technology need to be improved.

Case report: Ghost cell odontogenic carcinoma in a dog: diagnostics and surgical outcome.

A 6 year-old spayed female Poodle presented with a mandibular mass. Radiographic examination revealed osteolysis from the right mandibular canine to the fourth premolar, along with horizontal bone loss and dorsal displacement of the right mandibular first and second premolars. Skull cone beam computed tomography revealed osteolysis at the level of the right mandibular canine and fourth premolar. A destructive bone lesion was observed in the apical area of the right mandibular canine, with mass invasion of the interradicular bone of the right mandibular first molar near the mandibular canal. Consequently, unilateral total mandibulectomy and skin flap surgery were performed. Histopathological examination revealed poorly demarcated and infiltrative neoplastic epithelial cells that formed small islands and trabeculae. Neoplastic cells exhibited the malignant features of cytological atypia and high mitotic activity. Furthermore, the neoplastic epithelial cells frequently showed ghost cell changes and were diagnosed as ghost cell odontogenic carcinoma (GCOC). The dog was followed up for 1 year, during which no severe complications or local recurrence was observed, except for slight mandibular drift, tongue protrusion, and drooling. This case report describes the clinical features, diagnostic imaging, and histologic features of an unreported GCOC in a dog and the favorable outcome following surgical resection.

Novel Starter Strain Enterococcus faecium DMEA09 from Traditional Korean Fermented Meju.

The Enterococcus faecium strain DMEA09 was previously isolated from traditional Korean fermented meju. The objective of the current study was to investigate the traits of E. faecium strain DMEA09 as a starter candidate, focusing on its safety and technological properties. Regarding its safety, the DMEA09 strain was found to be sensitive to nine antibiotics (ampicillin, chloramphenicol, erythromycin, gentamicin, kanamycin, streptomycin, tetracycline, tylosin, and vancomycin) by showing lower minimum inhibitory concentrations (MICs) than the cut-off values suggested by the European Union Food Safety Authority for these nine antibiotics. However, its MIC value for clindamycin was twice as high as the cut-off value. A genomic analysis revealed that strain DMEA09 did not encode the acquired antibiotic resistance genes, including those for clindamycin. The DMEA09 strain did not show hemolysis as a result of analyzing α- and ß-hemolysis. It did not form biofilm either. A genomic analysis revealed that strain DMEA09 did not encode for any virulence factors including hemolysin. Most importantly, multilocus sequence typing revealed that the clonal group of strain DMEA09 was distinguished from clinical isolates. Regarding its technological properties, strain DMEA09 could grow in the presence of 6% salt. It showed protease activity when the salt concentration was 3%. It did not exhibit lipase activity. Its genome possessed 37 putative protease genes and salt-tolerance genes for survivability under salt conditions. Consequently, strain DMEA09 shows safe and technological properties as a new starter candidate. This was confirmed by genome analysis.

Staphylococcus equorum plasmid pKS1030-3 encodes auxiliary biofilm formation and trans-acting gene mobilization systems.

The foodborne bacterium Staphylococcus equorum strain KS1030 harbours plasmid pSELNU1, which encodes a lincomycin resistance gene. pSELNU1 undergoes horizontal transfer between bacterial strains, thus spreading antibiotic resistance. However, the genes required for horizontal plasmid transfer are not encoded in pSELNU1. Interestingly, a relaxase gene, a type of gene related to horizontal plasmid transfer, is encoded in another plasmid of S. equorum KS1030, pKS1030-3. The complete genome of pKS1030-3 is 13,583 bp long and encodes genes for plasmid replication, biofilm formation (the ica operon), and horizontal gene transfer. The replication system of pKS1030-3 possesses the replication protein-encoding gene repB, a double-stranded origin of replication, and two single-stranded origins of replication. The ica operon, relaxase gene, and a mobilization protein-encoding gene were detected in pKS1030-3 strain-specifically. When expressed in S. aureus RN4220, the ica operon and relaxase operon of pKS1030-3 conferred biofilm formation ability and horizontal gene transfer ability, respectively. The results of our analyses show that the horizontal transfer of pSELNU1 of S. equorum strain KS1030 depends on the relaxase encoded by pKS1030-3, which is therefore trans-acting. Genes encoded in pKS1030-3 contribute to important strain-specific properties of S. equorum KS1030. These results could contribute to preventing the horizontal transfer of antibiotic resistance genes in food.

ComQXPA quorum-sensing systems contribute to enhancing the protease activity of Bacillus velezensis DMB05 from fermented soybeans.

Bacillus velezensis DMB05, isolated from traditionally fermented soybean, meju, exhibited no protease activity on a TSA plate containing skim milk. To shed light on the genetic background behind this phenotypic non-protease activity, we analyzed the complete genome sequence of strain DMB05 and compared it with those of two B. velezensis strains which did exhibit protease activity. Comparative genome analyses showed no significant difference in the kind or number of proteases between the genomes of the three strains and that all strains possessed the degSU two-component system involved in the gene regulation of protease. However, strain DMB05 possessed a truncated comP which is part of the comQXPA operon that regulates the expression of degQ involved in the activation of DegSU. When the entire comQXPA operon derived from DMB06 was introduced into DMB05, the recombinant expressed proteolytic activity. The results of this experimental study provide evidence for the presence of regulatory genes involved in protease activity, one of several important factors involved in fermentation.

Performance enhancement of HfO2-based resistive random-access memory devices using ZnO nanoparticles.

In this study, resistive random-access memory (ReRAM) devices with ZnO nanoparticles (NPs) are suggested to enhance performance and reduce variation in device switching parameters. The ZnO NPs are formed by annealing ZnO prepared via atomic layer deposition on HfO2, which is verified using transmission electron microscopy, x-ray diffraction pattern, and atomic force microscopy. The depth profile analysis of x-ray photoelectron spectroscopy shows that oxygen diffuses from HfO2to ZnO NPs during annealing. This can be explained by the calculation results using density functional theory (DFT) where the formation energy of oxygen vacancies is reduced at the interface of ZnO NPs and HfO2compared to single HfO2. The fabricated ZnO NPs ReRAM demonstrates reduced forming voltage, stable resistive switching behavior, and improved cycle-to-cycle uniformity in a high-resistance state.

Analgesic effects and metabolome analyses of laser- and electro-acupuncture combined therapies in paclitaxel-induced neuropathic pain model.

Introduction: Allodynia, which can be induced by paclitaxel administration, is the presence of pain as a result of a stimulus that does not usually provoke pain. Many studies have investigated the analgesic efficacy of acupuncture, including laser acupuncture (LA) and electroacupuncture (EA). Although pain-related diseases are relatively common, few studies have analyzed the analgesic effects and mechanisms of LA combined with EA. The purpose of this study was to investigate the therapeutic effect and mechanism of manual acupuncture (MA), EA, LA, and combined therapy (LA + EA) in a paclitaxel-induced allodynia rat model. Methods: A total of 56 rats were classified into eight groups: a normal (Nor, n = 7), a control (Con, n = 7), an MA (n = 7), an EA (n = 7), a 650-nm LA (650LA, n = 7), an 830-nm LA (830LA, n = 7), a 650-nm LA combined with EA (650LA + EA, n = 7), and an 830-nm LA combined with EA group (830LA + EA, n = 7). Allodynia was induced by intraperitoneal injection of 2 mg/kg of paclitaxel every other day for a total of four times except the Nor group. Acupuncture treatments were conducted at the points of Jungwan (CV12) and Joksamni (ST36) once every other day for 6 min, for a total of nine times. Withdrawal response reaction times and force intensity of the foot were measured before the start of the experiment, after the 4th paclitaxel administration (day 8), and after the 9th and last treatment (day 15). On the 16th day, mRNA and protein expression in the spinal nerves was assessed, and a metabolome analysis of the animals' feces was performed. Results and discussion: Our analyses show that 650LA + EA treatment resulted in an upregulation of protein expression related to pain relief and nerve regeneration, whereas 830LA + EA treatment led to significant changes in metabolomes. This study demonstrates that a combination treatment of EA and LA can suppress allodynia and promote upregulation of protein expression related to nerve regeneration and is effective in changing the intestinal microbiome. Further large-scale research is required to assess the exact mechanism underlying the therapeutic effect of this combination treatment in pain-related diseases.

MTGEA: A Multimodal Two-Stream GNN Framework for Efficient Point Cloud and Skeleton Data Alignment.

Because of societal changes, human activity recognition, part of home care systems, has become increasingly important. Camera-based recognition is mainstream but has privacy concerns and is less accurate under dim lighting. In contrast, radar sensors do not record sensitive information, avoid the invasion of privacy, and work in poor lighting. However, the collected data are often sparse. To address this issue, we propose a novel Multimodal Two-stream GNN Framework for Efficient Point Cloud and Skeleton Data Alignment (MTGEA), which improves recognition accuracy through accurate skeletal features from Kinect models. We first collected two datasets using the mmWave radar and Kinect v4 sensors. Then, we used zero-padding, Gaussian Noise (GN), and Agglomerative Hierarchical Clustering (AHC) to increase the number of collected point clouds to 25 per frame to match the skeleton data. Second, we used Spatial Temporal Graph Convolutional Network (ST-GCN) architecture to acquire multimodal representations in the spatio-temporal domain focusing on skeletal features. Finally, we implemented an attention mechanism aligning the two multimodal features to capture the correlation between point clouds and skeleton data. The resulting model was evaluated empirically on human activity data and shown to improve human activity recognition with radar data only. All datasets and codes are available in our GitHub.

Increased Production of γ-Aminobutyric Acid from Brewer's Spent Grain Through Bacillus Fermentation.

Brewer's spent grain (BSG) is a waste product of the beer industry, and γ-aminobutyric acid (GABA) is a physiologically active substance important for brain and neuron physiology. In this study, we used the bacterial strains Bacillus velezensis DMB06 and B. licheniformis 0DA23-1, respectively, to ferment BSG and produce GABA. The GABA biosynthesis pathways were identified through genomic analysis of the genomes of both strains. We then inoculated the strains into BSG to determine changes in pH, acidity, reducing sugar content, amino-type nitrogen content, and GABA production, which was approximately doubled in BSG inoculated with Bacillus compared to that in uninoculated BSG; however, no significant difference was observed in GABA production between the two bacterial strains. These results provide the experimental basis for expanding the use of BSG by demonstrating the potential gain in increasing GABA production from a waste resource.

Comparison of four multilocus sequence typing schemes and amino acid biosynthesis based on genomic analysis of Bacillus subtilis.

Bacillus subtilis, a valuable industrial microorganism used in starter cultures in soybean fermentation, is a species of bacteria with interspecies diversity. Here, four multilocus sequence typing (MLST) schemes developed to assess the diversity of B. subtilis or Bacillus spp. were applied and compared to confirm the interspecies diversity of B. subtilis. In addition, we analyzed correlations between amino acid biosynthesis genes and sequence types (STs); this is important because amino acids are key taste components in fermented foods. On applying the four MLST methods to 38 strains and the type strain of B. subtilis, 30 to 32 STs were identified. The discriminatory power was 0.362-0.964 for the genes used in the MLST methods; the larger the gene, the greater the number of alleles and polymorphic sites. All four MLST methods showed a correlation between STs and strains that do not possess the hutHUIG operon (which contains genes required for the production of glutamate from histidine). This correlation was verified using 168 further genome-sequence strains.

Studies on Oxygen Permeation Resistance of SiCN Thin Film and RRAM Applications.

In this study, a silicon carbon nitride (SiCN) thin film was grown with a thickness of 5~70 nm by the plasma-enhanced chemical vapor deposition (PECVD) method, and the oxygen permeation characteristics were analyzed according to the partial pressure ratio (PPR) of tetramethylsilane (4MS) to the total gas amount during the film deposition. X-ray photoelectron spectroscopy (XPS), Fourier transform infrared spectroscopy (FT-IR), and X-ray reflectivity (XRR) were used to investigate the composition and bonding structures of the SiCN film. An atomic force microscope (AFM) was used to examine the surface morphology of the SiCN films to see the porosity. The analysis indicated that Si-N bonds were dominant in the SiCN films, and a higher carbon concentration made the film more porous. To evaluate the oxygen permeation, a highly accelerated temperature and humidity stress test (HAST) evaluation was performed. The films grown at a high 4MS PPR were more susceptible to oxygen penetration, which changed Si-N bonds to Si-N-O bonds during the HAST. These results indicate that increasing the 4MS PPR made the SiCN film more porous and containable for oxygen. As an application, for the first time, SiCN dielectric film is suggested to be applied to resistive random access memory (RRAM) as an oxygen reservoir to store oxygen and prevent a reaction between metal electrodes and oxygen. The endurance characteristics of RRAM are found to be enhanced by applying the SiCN.

A Study on the Resolution and Depth of Focus of ArF Immersion Photolithography.

In this study, the resolution and depth of focus (DOF) of the ArF immersion scanner are measured experimentally according to numerical aperture (NA). Based on the experiment, the theoretical trade-off relationship between the resolution and depth of focus can be confirmed and k1 and k2 are extracted to be about 0.288 and 0.745, respectively. Another observation for a problem in small critical dimension realization is the increase in line width roughness (LWR) according to mask open area ratio. To mitigate the trade-off problem and critical dimension variation, the photoresist thickness effect on depth of focus is analyzed. Generally, the photoresist thickness is chosen considering depth of focus, which is decided by NA. In practice, the depth of focus is found to be influenced by the photoresist thickness, which can be caused by the intensity change of the reflected ArF light. This means that photoresist thickness can be optimized under a fixed NA in ArF immersion photolithography technology according to the critical dimension and pattern density of the target layer.

Bottom-Gated ZnO TFT Pressure Sensor with 1D Nanorods.

In this study, a bottom-gated ZnO thin film transistor (TFT) pressure sensor with nanorods (NRs) is suggested. The NRs are formed on a planar channel of the TFT by hydrothermal synthesis for the mediators of pressure amplification. The fabricated devices show enhanced sensitivity by 16~20 times better than that of the thin film structure because NRs have a small pressure transmission area and causes more strain in the underlayered piezoelectric channel material. When making a sensor with a three-terminal structure, the leakage current in stand-by mode and optimal conductance state for pressure sensor is expected to be controlled by the gate voltage. A scanning electron microscope (SEM) was used to identify the nanorods grown by hydrothermal synthesis. X-ray diffraction (XRD) was used to compare ZnO crystallinity according to device structure and process conditions. To investigate the effect of NRs, channel mobility is also extracted experimentally and the lateral flow of current density is analyzed with simulation (COMSOL) showing that when the piezopotential due to polarization is formed vertically in the channel, the effective mobility is degraded.

Safety Assessment Systems for Microbial Starters Derived from Fermented Foods.

Microorganisms involved in food fermentation not only improve the aroma and taste of the food, but also enhance its preservation. Thus, they are added as starter cultures to boost the final product quality of commercial fermented foods. Although these microorganisms originate from fermented foods and have a long history of consumption, the European Union recently applied the concept of Qualified presumption of Safety (QPS), which is a safety evaluation system for microorganisms used in food or feed in Europe. The QPS system is a species-level safety system and shares results with the European Novel Food System, a strain-level safety evaluation system. In the United States, microorganisms added to fermented foods are considered as food additives or Generally Recognized as Safe substance. In Korea, food microbe lists are presented at the species level. Moreover, the nation has established a strain-oriented evaluation system that applies temporary safety evaluation methods for food raw materials as well as new raw materials. However, when it comes to microorganisms isolated from traditional fermented foods and other fermented food products, there is no definition of the term "species," and there is a lack of an evaluation system at the species level. Therefore, such an evaluation system for microbial species used in Korean fermented foods is necessary.

Transcriptomic analysis of Staphylococcus equorum KM1031 from the high-salt fermented seafood jeotgal under chloramphenicol, erythromycin and lincomycin stresses.

Staphylococcus equorum strain KM1031 is resistant to chloramphenicol, erythromycin and lincomycin. To shed light on the genetic factors underlying these antibiotic resistances, we determined the global gene expression profile of S. equorum KM1031 using RNA sequencing. During chloramphenicol, erythromycin and lincomycin treatment, 8.3% (183/2,336), 16.0% (354/2,336), and 2.9% (63/2,336) of S. equorum KM1031 genes exhibited significant differences in expression, respectively. These three antibiotics upregulated genes related to efflux and downregulated genes related to transporters. Antibiotic treatment also upregulated osmoprotectant-related genes involved in salt tolerance. To identify specific genes functionally related to antibiotic resistance, we compared the genome of strain KM1031 with those of three S. equorum strains that are sensitive to these three antibiotics. We identified three genes of particular interest: an antibiotic biosynthesis monooxygenase gene (abm, AWC34_RS01805) related to chloramphenicol resistance, an antibiotic ABC transporter ATP-binding protein gene (msr, AWC34_RS11115) related to erythromycin resistance, and a lincosamide nucleotydyltransferase gene (lnuA, AWC34_RS13300) related to lincomycin resistance. These genes were upregulated in response to the corresponding antibiotic; in particular, msr was upregulated more than fourfold by erythromycin treatment. Finally, the results of RNA sequencing were validated by quantitative real-time PCR. This transcriptomic analysis provides genetic evidence regarding antibiotic stress responses of S. equorum strain KM1031.

Case Report: Hindlimb Ataxia Concurrent With Seizures by Presumed Osmotic Demyelination Syndrome in a Dog.

A 6-year-old castrated male Chihuahua dog was presented with hindlimb paresis and ataxia. The dog had hyponatremia and was diagnosed as hypoadrenocorticism 10 days before its visit, and the neurologic signs including generalized tonic seizures and hindlimb paresis occurred 3 days after correction of hyponatremia at a referral hospital. Based on history and clinical findings, osmotic demyelination syndrome (ODS) secondary to rapid correction of hyponatremia was highly suspected. After administration of anti-convulsant and supplements, seizures did not occur, and gait was normalized within 2 weeks. Phenobarbital was tapered and finally discontinued after 3 months, and seizure did not recur. The neurologic signs were completely resolved and the dog continued to be free of neurologic or additional clinical signs over the 19-month follow-up period. ODS should be included among the differential diagnoses in case of any acute neurological dysfunction that occurs with episodes of rapid correction of hyponatremia. To the author's knowledge, this is the rare case report of a dog with hypoadrenocorticism and presumed ODS after rapid correction of hyponatremia leading to neurologic signs including seizures and ataxia.

Discrimination of Bacillus subtilis from Other Bacillus Species Using Specific Oligonucleotide Primers for the Pyruvate Carboxylase and Shikimate Dehydrogenase Genes.

Bacillus subtilis is a useful bacterium in the food industry with applications as a starter strain for fermented food and as a probiotic. However, it is difficult to discriminate B. subtilis from other Bacillus species because of high phenotypic and genetic similarity. In this study, we employed five previously constructed multilocus sequence typing (MLST) methods for the discrimination of B. subtilis from other Bacillus species and all five MLST assays clearly distinguished B. subtilis. Additionally, the 17 housekeeping genes used in the five MLST assays also clearly distinguished B. subtilis. The pyruvate carboxylase (pyrA) and shikimate dehydrogenase (aroE) genes were selected for the discrimination of B. subtilis because of their high number of polymorphic sites and the fact that they displayed the lowest homology among the 17 housekeeping genes. Specific primer sets for the pyrA and aroE genes were designed and PCR products were specifically amplified from B. subtilis, demonstrating the high specificity of the two housekeeping genes for B. subtilis. This species-specific PCR method provides a quick, simple, powerful, and reliable alternative to conventional methods in the detection and identification of B. subtilis.

Dynamic Task Offloading for Cloud-Assisted Vehicular Edge Computing Networks: A Non-Cooperative Game Theoretic Approach.

Vehicular edge computing (VEC) is one of the prominent ideas to enhance the computation and storage capabilities of vehicular networks (VNs) through task offloading. In VEC, the resource-constrained vehicles offload their computing tasks to the local road-side units (RSUs) for rapid computation. However, due to the high mobility of vehicles and the overloaded problem, VEC experiences a great deal of challenges when determining a location for processing the offloaded task in real time. As a result, this degrades the quality of vehicular performance. Therefore, to deal with these above-mentioned challenges, an efficient dynamic task offloading approach based on a non-cooperative game (NGTO) is proposed in this study. In the NGTO approach, each vehicle can make its own strategy on whether a task is offloaded to a multi-access edge computing (MEC) server or a cloud server to maximize its benefits. Our proposed strategy can dynamically adjust the task-offloading probability to acquire the maximum utility for each vehicle. However, we used a best response offloading strategy algorithm for the task-offloading game in order to achieve a unique and stable equilibrium. Numerous simulation experiments affirm that our proposed scheme fulfills the performance guarantees and can reduce the response time and task-failure rate by almost 47.6% and 54.6%, respectively, when compared with the local RSU computing (LRC) scheme. Moreover, the reduced rates are approximately 32.6% and 39.7%, respectively, when compared with a random offloading scheme, and approximately 26.5% and 28.4%, respectively, when compared with a collaborative offloading scheme.