Isolation, characterization, and application of bacteriophage on Vibrio parahaemolyticus biofilm to control seafood contamination.

OBJECTIVE: This study intended to isolate a Vibrio-particular phage from the natural environment, analyse its characteristics and genome sequence, and investigate its reduction effect on V. parahaemolyticus biofilm as a biocontrol agent in squid and mackerel. METHODS: Among 21 phages, phage CAU_VPP01, isolated from beach mud, was chosen for further experiments based on host range and EOP tests. When examining the reduction effect of phage CAU_VPP01 against Vibrio parahaemolyticus biofilms on surfaces (stainless steel [SS] and polyethylene terephthalate [PET]) and food surfaces (squid and mackerel). RESULTS: The phage showed the most excellent reduction effect at a multiplicity-of-infection (MOI) 10. Three-dimensional images acquired with confocal laser scanning microscopy (CLSM) analysis were quantified using COMSTAT, which showed that biomass, average thickness, and roughness coefficient decreased when treated with the phage. Colour and texture analysis confirmed that the quality of squid and mackerel was maintained after the phage treatment. Finally, a comparison of gene expression levels determined by qRT-PCR analysis showed that the phage treatment induced a decrease in the gene expression of flaA, vp0962, andluxS, as examples. CONCLUSION: This study indicated that Vibrio-specific phage CAU_VPP01 effectively controlled V. parahaemolyticus biofilms under various conditions and confirmed that the isolated phage could possibly be used as an effective biocontrol weapon in the seafood manufacturing industry.

Correlation between the Leaflet-Annulus Index and Echocardiographic Indices in Maltese Dogs with Myxomatous Mitral Valve Disease.

Myxomatous mitral valve disease (MMVD) is the most common chronic heart valve disease, leading to left-sided cardiomegaly in dogs. The leaflet-annulus index (LAI) was originally used in humans as a predictor of mitral regurgitation (MR) after mitral valve repair surgery. This index represents the quantity and severity of MR since it is affected by annular dilation. Recently, LAI was adapted to veterinary medicine, and its usefulness as an indicator of annular dilation on 2D transthoracic echocardiography in MMVD dogs was suggested. For this study, 135 Maltese dogs were selected and divided into groups of control, B1, and B2, according to the American College of Veterinary Internal Medicine consensus statement. The following data were collected: radiographic indices including the vertebral heart score and vertebral left atrial size, echocardiographic indices including the left-atrium-to-aortic-root ratio (LA:Ao), left ventricular internal diameter at diastole, normalized for body weight, and anteroposterior length and LAI measured on right parasternal long-axis view. The results showed a significant difference in LAI between each group, becoming smaller as the disease progressed. Also, there was a significant correlation between LAI and each index, showing the strongest correlation with LA:Ao. LAI could be helpful as a new indicator used for the determination of severity and prognosis in Maltese dogs with MMVD.

Scheduling Diagnostic Testing Kit Deliveries with the Mothership and Drone Routing Problem.

A critical component in the public health response to pandemics is the ability to determine the spread of diseases via diagnostic testing kits. Currently, diagnostic testing kits, treatments, and vaccines for the COVID-19 pandemic have been developed and are being distributed to communities worldwide, but the spread of the disease persists. In conjunction, a strong level of social distancing has been established as one of the most basic and reliable ways to mitigate disease spread. If home testing kits are safely and quickly delivered to a patient, this has the potential to significantly reduce human contact and reduce disease spread before, during, and after diagnosis. This paper proposes a diagnostic testing kit delivery scheduling approach using the Mothership and Drone Routing Problem (MDRP) with one truck and multiple drones. Due to the complexity of solving the MDRP, the problem is decomposed into 1) truck scheduling to carry the drones and 2) drone scheduling for actual delivery. The truck schedule (TS) is optimized first to minimize the total travel distance to cover patients. Then, the drone flight schedule is optimized to minimize the total delivery time. These two steps are repeated until it reaches a solution minimizing the total delivery time for all patients. Heuristic algorithms are developed to further improve the computational time of the proposed model. Experiments are made to show the benefits of the proposed approach compared to the commonly performed face-to-face diagnosis via the drive-through testing sites. The proposed solution method significantly reduced the computation time for solving the optimization model (less than 50 minutes) compared to the exact solution method that took more than 10 hours to reach a 20% optimality gap. A modified basic reproduction rate (i.e., m R 0) is used to compare the performance of the drone-based testing kit delivery method to the face-to-face diagnostic method in reducing disease spread. The results show that our proposed method (m R 0= 0.002) outperformed the face-to-face diagnostic method (m R 0= 0.0153) by reducing m R 0 by 7.5 times.

Superficial intense focused ultrasound on periorbital wrinkle.

The periorbital wrinkles are easily perceived evidence of aging, so become a major concern for many patients. Various treatments have been attempted to improve periorbital wrinkles, but the need for new treatments that are less invasive and more effective is still high. In this study, we evaluated the safety, clinical and histological effects of intense focused ultrasound using only a 1.5 mm transducer in the management of periorbital wrinkles. Ten adult Korean females were enrolled. The treatment effect and safety profile were evaluated up to 3 months after 1 session of IFUS treatment on the periorbital wrinkles. The mean subjective satisfaction score was 3.2 ± 0.79 (mean ± standard deviation) by 5- point scale. The mean objective clinical improvement score was highest in the fine wrinkle on the crow's feet area and lowest in the deep wrinkles of the infraorbital and crow's feet area. Histometrically, increase of collagen and elastic fiber density was observed in the all layers of dermis. No serious side effects occurred after the treatment. In conclusion, intense focused ultrasound treatment using a 1.5-mm transducer alone can significantly improve periocular wrinkles after a single treatment with a good safety profile.

Metabolomic approaches to polyamines including acetylated derivatives in lung tissue of mice with asthma.

INTRODUCTION: Recently, the relationship between polyamine (PA) metabolism and asthma has been studied in severe asthmatic therapy, but systematic PA metabolism including their acetylated derivatives was not fully understood. OBJECTIVES: Profiling analysis of polyamines (PAs) was performed to understand the biochemical events and monitor altered PA metabolism in lung tissue of mice with asthma. METHODS: Polyamine profiling of lung tissue of mice with asthma was performed without derivatization by liquid chromatography-tandem mass spectrometry (LC-MS/MS) combined with star pattern recognition analysis. The PA levels between control and asthma groups were evaluated by multivariate analysis. RESULTS: In mouse lung tissue, seven PAs were determined by LC-MS/MS in multiple reaction monitoring (MRM) mode. Their levels were normalized to the corresponding mean levels of the control group for star pattern analysis, which showed distorted heptagonal shapes with characteristic and readily distinguishable patterns for each group. Levels of putrescine (p < 0.0034), N1-acetylputrescine (p < 0.0652), and N8-acetylspermidine (p < 0.0827) were significantly increased in asthmatic lung tissue. The separation of the two groups was evaluated using multivariate analysis. In unsupervised learning, acetylated PAs including N1-acetylspermine were the main metabolites for discrimination. In supervised learning, putrescine and N1-acetylputrescine were evaluated as important metabolites. CONCLUSIONS: The present results provide basic data for understanding polyamine metabolism in asthma and may help to improve the therapy for severe asthma patients.

Retraction Note to: Analysis of loxoprofen in tablets, patches, and equine urine as tert-butyldimethylsilyl derivative by gas chromatography-mass spectrometry.

The authors have retracted this article [1] because after publication they became aware that the equine urine samples analysed for loxoprofen in this study were in fact equine plasma samples. Therefore the results and conclusions of this article cannot be relied upon. All authors agree to this retraction.

Identification of a closed cutaneous injury after mechanical trauma caused by collision.

PURPOSE: Robotics has evolved rapidly in terms of mechanical design and control in the past few years. Collaborative robots that have direct contact with humans are being introduced in various fields, including industrial and medical services. Because collaborative robot systems are being introduced rapidly, the safety of the humans who work with them is becoming an important issue. In this study, we investigated skin injuries resulting from a collision between robots and humans using a freefall experiment system. METHODS: We particularly focused on closed skin injuries caused by a collision. To induce a closed injury, we struck mini-pigs with cubic-edge square and semi-sphere impactors at collision speeds of 1 and 3 m/s. We did not observe any open injuries with those conditions. Closed injuries were observed in the dermal layer of the skin after the collision test at both speeds and with both impactors. RESULTS: The collagen fiber in the dermal layer was separated and fragmented, and the subcutaneous fat layer became dense as a result of the collision. CONCLUSIONS: We closely observed and analyzed the histopathologic changes in the dermal and subcutaneous layers with intact epidermis after mechanical trauma to the inner skin layers.

Safety of Temporary Use of Recycled Autoclaved Femoral Components in Infected Total Knee Arthroplasty: Confirming Sterility Using a Sonication Method.

BACKGROUND: The purpose of this study was to evaluate the usefulness of sonication technique for microbiological diagnosis and the sterility of the recycled autoclaved femoral components from infected total knee arthroplasty (TKA) using a sonication method. METHODS: Nineteen femoral implants explanted from patients with infected TKA were sterilized with a standard autoclave method. Standard culture of the fluid before and after sonication of the sterilized implants was performed to detect pathogenic microorganisms. Additional experiments were performed to evaluate the sterility of the recycled implant by inducing artificial biofilm formation. Methicillin-resistant Staphylococcus aureus (MRSA) was inoculated into 10 implants and sterilization in a standard autoclave was performed, and then the fluid was cultured before and after sonication. RESULTS: Two of the 19 sterilized implants were positive for growth of bacteria after sonication, whereas no growth was detected in the cultured fluid from the sterilized implants before sonication. The bacteria were Staphylococcus species in all two cases. In one of 10 implants inoculated with MRSA, the culture was positive for growth of bacteria both before and after sonication. However, Staphylococcus epidermidis was cultured from both occasions and thus this implant was thought to be contaminated. CONCLUSIONS: We found sonication for identification of pathogens could be helpful, but this finding should be interpreted carefully because of the possibility of contamination. Sterilization of an infected femoral implant with an autoclave method could be a good method for using the temporary articulating antibiotic spacer in two-stage revision arthroplasty.

Analysis of loxoprofen in tablets, patches, and equine urine as tert-butyldimethylsilyl derivative by gas chromatography-mass spectrometry.

Loxoprofen is a non-steroidal anti-inflammatory drug of the 2-arylpropionic acid type, which has used to treat musculoskeletal disorders in the horse racing industry. However, it has also used illicitly to mask clinical signs of inflammation and pain in racehorses. Thus, its accurate analysis has become an important issue in horse doping laboratories. In this study, an analytical method of loxoprofen was developed as tert-butyldimethylsilyl (TBDMS) derivative by gas chromatography-mass spectrometry (GC-MS). Characteristic fragment ions of [M-15], [M-57], and [M-139] permitted the accurate and selective detection of loxoprofen. Under optimal conditions, this method showed good linearity (r ≥ 0.999) in the range of 10-500 ng/mL, repeatability (% relative standard deviation = 5.6-8.5), and accuracy (% relative error = - 0.3-0.9) with a detection limit of 1.0 ng. When applied to the analysis of loxoprofen in tablet and patch products, loxoprofen was positively identified as TBDMS derivative by GC-MS. The present method provided rapid and accurate determination of loxoprofen in patch and tablet products. Levels of loxoprofen were highest in equine urine at 0.5 and 1 h after oral administration with single dose (3 mg/kg) to three horses, and then rapidly reduced to below the lower limit of quantification at 24 h. Therefore, the present method will be useful for the pharmacokinetic study and doping tests for loxoprofen and other similar acidic drugs in horses.

Integration of metabolomics and transcriptomics in nanotoxicity studies.

Biomedical research involving nanoparticles has produced useful products with medical applications. However, the potential toxicity of nanoparticles in biofluids, cells, tissues, and organisms is a major challenge. The '-omics' analyses provide molecular profiles of multifactorial biological systems instead of focusing on a single molecule. The 'omics' approaches are necessary to evaluate nanotoxicity because classical methods for the detection of nanotoxicity have limited ability in detecting miniscule variations within a cell and do not accurately reflect the actual levels of nanotoxicity. In addition, the 'omics' approaches allow analyses of in-depth changes and compensate for the differences associated with high-throughput technologies between actual nanotoxicity and results from traditional cytotoxic evaluations. However, compared with a single omics approach, integrated omics provides precise and sensitive information by integrating complex biological conditions. Thus, these technologies contribute to extended safety evaluations of nanotoxicity and allow the accurate diagnoses of diseases far earlier than was once possible in the nanotechnology era. Here, we review a novel approach for evaluating nanotoxicity by integrating metabolomics with metabolomic profiling and transcriptomics, which is termed "metabotranscriptomics". [BMB Reports 2018; 51(1): 14-20].

Analysis of the free fatty acid metabolome in the plasma of patients with systemic lupus erythematosus and fever.

INTRODUCTION: Systemic lupus erythematosus (SLE) is a multifactorial autoimmune disease with heterogeneous clinical manifestations mediated by immune dysregulation. OBJECTIVES: We aimed to analyze the metabolomic differences in free fatty acids (FFAs) between patients with SLE and healthy controls (HCs). METHODS: In this study, the levels of 24 FFAs, as their tert-butyldimethylsilyl derivatives, in the plasma of 41 patients with SLE and 41 HCs, were investigated using gas chromatography with mass spectrometry in selected-ion monitoring mode. RESULTS: The results showed that patients with SLE and HCs had significantly different levels of 13 of the 24 FFAs. The levels of myristic, palmitoleic, oleic, and eicosenoic acids were significantly higher, whereas the levels of caproic, caprylic, linoleic, stearic, arachidonic, eicosanoic, behenic, lignoceric, and hexacosanoic acids were significantly lower in patients with SLE, than in the HCs. In the partial-correlation analysis of the FFA profiles and markers of disease activity of SLE, several metabolic markers correlated with SLE disease activity. CONCLUSIONS: Our results provide a comprehensive understanding of the relationship between FFAs and markers of SLE disease activity. Thus, this approach has promising potential for the discovery of metabolic biomarkers of SLE.

Comparison of Fecal Microbiota between German Holstein Dairy Cows with and without Left-Sided Displacement of the Abomasum.

One of the most common diseases in high-performance German Holstein dairy cows is left-sided displacement of the abomasum (LDA). Hypomotility of the abomasum is detrimental during the pathogenesis of LDA. It is known that improper interactions between the gut microbiota and the enteric nervous system contribute to dysfunctions of gastrointestinal motility. Therefore, we hypothesized that the gut microbial composition will be different between German Holstein dairy cows with and without LDA. We used 16S rRNA gene analysis to evaluate whether there are any differences in bacterial composition between German Holstein dairy cows with and without LDA. Even though our data are limited to being used to correlate compositional changes with corresponding functional aspects in the pathogenesis of LDA, results from this study show that the fecal microbial compositions of German Holstein dairy cows with LDA shifted and were less diverse than those in normal cows. In particular,Spirochaeteswere absent in cows with LDA.

Molecular Detection of Giardia intestinalis from Stray Dogs in Animal Shelters of Gyeongsangbuk-do (Province) and Daejeon, Korea.

Giardia is a major public health concern and considered as reemerging in industrialized countries. The present study investigated the prevalence of giardiosis in 202 sheltered dogs using PCR. The infection rate was 33.2% (67/202); Gyeongsangbuk-do and Daejeon showed 25.7% (39/152, P<0.0001) and 56% (28/50), respectively. The prevalence of infected female dogs (46.7%, P<0.001) was higher than in male dogs (21.8%). A higher prevalence (43.5%, P<0.0001) was observed in mixed breed dogs than purebred (14.1%). Although most of the fecal samples collected were from dogs of ≥1 year of age which showed only 27.4% positive rate, 61.8% (P<0.001) of the total samples collected from young animals (<1 year of age) were positive for G. intestinalis. A significantly higher prevalence in symptomatic dogs (60.8%, P<0.0001) was observed than in asymptomatic dogs (23.8%). Furthermore, the analysis of nucleotide sequences of the samples revealed that G. intestinalis Assemblages A and C were found in the feces of dogs from Gyeongsangbuk-do and Daejeon. Since G. intestinalis Assemblage A has been known to infect humans, our results suggest that dogs can act as an important reservoir of giardiosis in Korea. Hence, hygienic management should be given to prevent possible transmission to humans.

Application of vincristine-loaded platelet therapy in three dogs with refractory immune-mediated thrombocytopenia.

Three dogs presented with refractory immune-mediated thrombocytopenia (IMT). All patients failed to respond to prednisone, which is considered a mainstay of immunosuppressive therapy. Vincristine-loaded platelets (VLPs), which act selectively on mononuclear phagocytes,were introduced. After the VLPs were transfused, two dogs responded quickly with improved clinical signs while the third dog with recurrent IMT was euthanized due to its deteriorating condition. This case report describes the efficacy of VLP therapy in refractory IMT patients.