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Bovine viral diarrhea (BVD) is a single-stranded, positive-sense ribonucleic acid (RNA) virus belonging to the genus Pestivirus of the Flaviviridae family. BVD frequently causes economic losses to farmers. Among bovine viral diarrhea virus (BVDV) strains, BVDV-1b is predominant and widespread in Hanwoo calves. Reverse-transcription polymerase chain reaction (RT-PCR) is an essential method for diagnosing BVDV-1b and has become the gold standard for diagnosis in the Republic of Korea. However, this diagnostic method is time-consuming and requires expensive equipment. Therefore, Clustered regularly interspaced short palindromic repeats-Cas (CRISPR-Cas) systems have been used for point-of-care (POC) testing of viruses. Developing a sensitive and specific method for POC testing of BVDV-1b would be advantageous for controlling the spread of infection. Thus, this study aimed to develop a novel nucleic acid detection method using the CRISPR-Cas13 system for POC testing of BVDV-1b. The sequence of the BVD virus was extracted from National Center for Biotechnology Information (NC_001461.1), and the 5' untranslated region, commonly used for detection, was selected. CRISPR RNA (crRNA) was designed using the Cas13 design program and optimized for the expression and purification of the LwCas13a protein. Madin Darby bovine kidney (MDBK) cells were infected with BVDV-1b, incubated, and the viral RNA was extracted. To enable POC viral detection, the compatibility of the CRISPR-Cas13 system was verified with a paper-based strip through collateral cleavage activity. Finally, a colorimetric assay was used to evaluate the detection of BVDV-1b by combining the previously obtained crRNA and Cas13a protein on a paper strip. In conclusion, the CRISPR-Cas13 system is highly sensitive, specific, and capable of nucleic acid detection, making it an optimal system for the early point-of-care testing of BVDV-1b.
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In this study, a static mixer was used as an alternative to the existing flash mixing method for ballasted flocculation to assess the turbidity removal and ballasted floc formation characteristics. Synthetic magnetite exhibits excellent properties, such as high specific gravity, hydrophobicity, and wear resistance, making it a suitable ballast agent (BA). The experimental design was optimized using the response surface methodology. To evaluate turbidity removal, a model based on polyaluminum chloride dosage, BA surface charge, and pH was developed. To assess the ballasted floc characteristics, the BA dosage, BA size, and G value of the static mixer were used. During ballasted flocculation, the impact of the zeta potential of the BA was minimal. Consequently, bonding primarily resulted from the viscosity of the floc caused by physical collisions rather than electrostatic forces stemming from the BA charge. The findings of this study demonstrated promising outcomes, including potential energy savings and process streamlining, by identifying crucial design elements for implementing a static mixer in the ballasted flocculation process.
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Óxido Ferroso-Férrico , Floculação , Óxido Ferroso-Férrico/química , Purificação da Água/métodos , Hidróxido de Alumínio/química , Interações Hidrofóbicas e Hidrofílicas , Eliminação de Resíduos Líquidos/métodos , Concentração de Íons de HidrogênioRESUMO
The material transport system, facilitated by motor proteins, plays a vital role in maintaining a non-equilibrium cellular state. However, understanding the temporal coordination of motor protein activity requires an advanced imaging technique capable of measuring 3D angular displacement in real-time. In this study, a Fourier transform-based plasmonic dark-field microscope has been developed using anisotropic nanoparticles, enabling the prolonged and simultaneous observation of endosomal lateral and rotational motion. A sequence of discontinuous 3D angular displacements has been observed during the pause and run phases of transport. Notably, a serially correlated temporal pattern in the intermittent rotational events has been demonstrated during the tug-of-war mechanism, indicating Markovian switching between the exploitational and explorational modes of motor protein exchange prior to resuming movement. Alterations in transition frequency and the exploitation-to-exploration ratio upon dynein inhibitor treatment highlight the relationship between disrupted motor coordination and reduced endosomal transport efficiency. Collectively, these results suggest the importance of orchestrated temporal motor protein patterns for efficient cellular transport.
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Endossomos , Endossomos/metabolismo , Humanos , Microscopia/métodos , Dineínas/metabolismo , Transporte Biológico/fisiologia , Proteínas Motores Moleculares/metabolismoRESUMO
Understanding the spatial organization of membrane proteins is crucial for unraveling key principles in cell biology. The reaction-diffusion model is commonly used to understand biochemical patterning; however, applying reaction-diffusion models to subcellular phenomena is challenging because of the difficulty in measuring protein diffusivity and interaction kinetics in the living cell. In this work, we investigated the self-organization of the plasmalemma vesicle-associated protein (PLVAP), which creates regular arrangements of fenestrated ultrastructures, using single-molecule tracking. We demonstrated that the spatial organization of the ultrastructures is associated with a decrease in the association rate by actin destabilization. We also constructed a reaction-diffusion model that accurately generates a hexagonal array with the same 130 nm spacing as the actual scale and informs the stoichiometry of the ultrastructure, which can be discerned only through electron microscopy. Through this study, we integrated single-molecule experiments and reaction-diffusion modeling to surpass the limitations of static imaging tools and proposed emergent properties of the PLVAP ultrastructure.
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Proteínas de Transporte , Proteínas de Membrana , Proteínas de Membrana/metabolismo , Difusão , Modelos BiológicosRESUMO
OBJECTIVE: To review the characteristics of distal middle cerebral artery (MCA) aneurysm treated by microsurgery, the detailed surgical options, and the clinical result. METHODS: We retrospectively reviewed cerebral aneurysm in the M2 and M3 segments of the MCA surgically treated between January 2015 and December 2022. The demographic data, aneurysm-related findings, type of surgical approach, surgical technique, and clinical outcomes of the enrolled patients were analyzed. RESULTS: Sixteen distal MCA aneurysms were treated with microneurosurgery (incidence, 1.0%; female, 12; mean age, 58.1 years; ruptured, three). Twelve aneurysms were in the M2 segment (insular segment), two aneurysms at the M2-M3 junction, and two aneurysms in the M3 segment (opercular segment). Twelve aneurysms were saccular (average size, 4.9 mm; multiplicity, 50%; average aneurysms, 3.0; partially thrombosed, 1; sidewall aneurysm, 2). Three aneurysms were fusiform, of which two were ruptured. Of the ruptured aneurysms, one was a ruptured dissecting aneurysm. The trans-sylvian and trans-sulcal approaches were used in fourteen and two patients, respectively. Neck clipping, wrap clipping, and surgical trapping were performed in twelve, one, and one patient, respectively. Proximal occlusion was performed in one patient. Bypass technique was required in two patients (neck clipping and proximal occlusion). The modified Rankin Score was 6 in the two patients with ruptured aneurysms. The remaining patients did not show further neurological deterioration after microneurosurgery. CONCLUSIONS: Distal MCA aneurysms had a high incidence of being diagnosed with multiple other aneurysms and were relatively non-saccular.
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The biological process of aging is thought to result in part from accumulation of senescent cells in organs. However, the present study identified a subset of fibroblasts and smooth muscle cells which are the major constituents of organ stroma neither proliferative nor senescent in tissues of the elderly, which we termed "mid-old status" cells. Upregulation of pro-inflammatory genes (IL1B and SAA1) and downregulation of anti-inflammatory genes (SLIT2 and CXCL12) were detected in mid-old cells. In the stroma, SAA1 promotes development of the inflammatory microenvironment via upregulation of MMP9, which decreases the stability of epithelial cells present on the basement membrane, decreasing epithelial cell function. Remarkably, the microenvironmental change and the functional decline of mid-old cells could be reversed by a young cell-originated protein, SLIT2. Our data identify functional reversion of mid-old cells as a potential method to prevent or ameliorate aspects of aging-related tissue dysfunction.
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Envelhecimento , Senescência Celular , Humanos , Idoso , Senescência Celular/genética , Envelhecimento/genética , Células Epiteliais/fisiologia , Fibroblastos , Miócitos de Músculo LisoRESUMO
Catastrophic forgetting, which means a rapid forgetting of learned representations while learning new data/samples, is one of the main problems of deep neural networks. In this paper, we propose a novel incremental learning framework that can address the forgetting problem by learning new incoming data in an online manner. We develop a new incremental learning framework that can learn extra data or new classes with less catastrophic forgetting. We adopt the hippocampal memory process to the deep neural networks by defining the effective maximum of neural activation and its boundary to represent a feature distribution. In addition, we incorporate incremental QR factorization into the deep neural networks to learn new data with both existing labels and new labels with less forgetting. The QR factorization can provide the accurate subspace prior, and incremental QR factorization can reasonably express the collaboration between new data with both existing classes and new class with less forgetting. In our framework, a set of appropriate features (i.e., nodes) provides improved representation for each class. We apply our method to the convolutional neural network (CNN) for learning Cifar-100 and Cifar-10 datasets. The experimental results show that the proposed method efficiently alleviates the stability and plasticity dilemma in the deep neural networks by providing the performance stability of a trained network while effectively learning unseen data and additional new classes.
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In live cells, the plasma membrane is composed of lipid domains separated by hundreds of nanometers in dynamic equilibrium. Lipid phase separation regulates the trafficking and spatiotemporal organization of membrane molecules that promote signal transduction. However, visualizing domains with adequate spatiotemporal accuracy remains challenging because of their subdiffraction limit size and highly dynamic properties. Here, we present a single lipid-molecular motion analysis pipeline (lipid-MAP) for analyzing the phase heterogeneity of lipid membranes by detecting the instantaneous velocity change of a single lipid molecule using the excellent optical properties of nanoparticles, high spatial localization accuracy of single-molecule localization microscopy, and separation capability of the diffusion state of the hidden Markov model algorithm. Using lipid-MAP, individual lipid molecules were found to be in dynamic equilibrium between two statistically distinguishable phases, leading to the formation of small (â¼170 nm), viscous (2.5× more viscous than surrounding areas), and transient domains in live cells. Moreover, our findings provide an understanding of how membrane compositional changes, i.e., cholesterol and phospholipids, affect domain formation. This imaging method can contribute to an improved understanding of spatiotemporal-controlled membrane dynamics at the molecular level.
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Fosfolipídeos , Transdução de Sinais , Membrana Celular/metabolismo , Fosfolipídeos/metabolismo , Membranas , Difusão , Bicamadas Lipídicas/metabolismoRESUMO
OBJECTIVE: The real-world INFORM study analyzed sociodemographics, treatment patterns and clinical outcomes for patients with newly diagnosed advanced epithelial ovarian cancer (EOC) in Australia, South Korea (S.Korea) and Taiwan preceding incorporation of poly(ADP-ribose) polymerase inhibitors into clinical practice. METHODS: Retrospective data from patients diagnosed with EOC (high-grade serous EOC for Taiwan) between January 2014 and December 2018 with ≥12 months follow-up from diagnosis were analyzed descriptively. Survival was evaluated by Kaplan-Meier with two-sided 95% confidence interval (CI). RESULTS: Of the 987 patients (Australia, 223; S.Korea, 513; Taiwan, 251), 98% received platinum-based chemotherapy (CT). In S.Korea and Taiwan 76.0% and 78.9% respectively underwent primary cytoreductive surgery; in Australia, 56.5% had interval debulking surgery. Bevacizumab was included in primary/maintenance therapy for 22.4%, 14.6% and 6.8% of patients in Australia, S.Korea and Taiwan, respectively. Patients receiving bevacizumab were high-risk (reimbursement policy) and achieved similar real-world progression-free survival (PFS) compared with CT only. Overall, the median real-world PFS (months; 95% CI) was similar across Australia (16.0 [14.63-18.08]), S.Korea (17.7 [16.18-19.27]) and Taiwan (19.1 [17.56-22.29]). CONCLUSION: This study reveals poor prognosis despite differences in demographics and treatment patterns for patients with EOC across Asia-Pacific suggesting the need for biomarker-driven novel therapies to improve outcomes.
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Neoplasias Ovarianas , Humanos , Feminino , Carcinoma Epitelial do Ovário/tratamento farmacológico , Neoplasias Ovarianas/terapia , Neoplasias Ovarianas/tratamento farmacológico , Bevacizumab/uso terapêutico , Taiwan/epidemiologia , Estudos Retrospectivos , Terapia Neoadjuvante , República da Coreia/epidemiologia , Inibidores de Poli(ADP-Ribose) Polimerases/uso terapêutico , Procedimentos Cirúrgicos de CitorreduçãoRESUMO
With the development of maritime technology and equipment, most ships are equipped with an automatic identification system (AIS) to store navigation information. Over time, the size of the data increases, rendering its storage and processing difficult. Hence, it is necessary to transform the AIS data into trajectories, and then simplify the AIS trajectories to remove unnecessary information that is not related to route shape. Moreover, topographic information must be considered because otherwise, the simplified trajectory can intersect obstacles. In this study, we propose an AIS trajectory simplification algorithm considering topographic information. The proposed algorithm simplifies the trajectories without the intersection of the trajectory and obstacle using the improved Douglas-Peucker algorithm. Polygon map random (PMR) quadtree was used to consider topographic information on the coast, and the intersection between topographic information and simplified trajectories was efficiently computed using the PMR quadtree. To verify the effectiveness of the proposed algorithm, experiments were conducted on real-world trajectories in the Korean sea. The proposed algorithm yielded simplified trajectories with no intersections of the trajectory and obstacle. In addition, the computational efficiency of the proposed algorithm with the PMR quadtree was superior to that without the PMR quadtree.
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Objective: To assess risk factors for encephalopathy in non-ICU hospitalized patients with COVID-19 and the effect of encephalopathy on short-term outcomes. Methods: We collected clinical and electrophysiological characteristics of fifty patients with COVID-19 infection admitted to a ward service and who had an electroencephalogram (EEG) performed. Associations with short-term outcomes including hospital length of stay and discharge disposition were determined from univariate and multivariate statistical analysis. Results: Clinical delirium was associated with encephalopathy on EEG, cefepime use was associated with increased length of stay, and of all factors analyzed, background frequency on EEG alone was correlated with discharge disposition. Conclusion: Encephalopathy is one of the major determinants of short-term outcomes in hospitalized non-ICU patients with COVID-19.
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Live video recording of intracellular material transport is a promising means of deciphering the fascinating underlying mechanisms driving life at the molecular level. Such technology holds the key to realizing real-time observation at appropriate resolutions in three-dimensional (3D) space within living cells. Here, we report an optical microscopic method for probing endosomal dynamics with proper spatiotemporal resolution within 3D space in live cells: plasmonic dark-field STORM (pdf-STORM). We first confirmed that pdf-STORM has a spatial resolution comparable to that of scanning electron microscopy. Additionally, by observing two optical probes within a single organelle, we were able to track rotational movements and demonstrate the feasibility of using pdf-STORM to observe the angular displacements of an endosome during a "tug-of-war" over an extended period. Finally, we show various biophysical parameters of the hitherto unelucidated dynamics of endosomes-angular displacement is discontinuous and y-axis movement predominates and follows a long-tail distribution.
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The CO2 atmospheric concentration level hit the record at more than 400 ppm and is predicted to keep increasing as the dependence on fossil fuels is inevitable. The CO2 electrocatalytic conversion becomes an alternative due to its environmental and energy-friendly properties and benign operation condition. Lately, bimetallic materials have drawn significant interest as electrocatalysts due to their distinct properties, which the parents' metal cannot mimic. Herein, the indium-bismuth nanosphere (In16Bi84 NS) was fabricated via the facile liquid-polyol technique. The In16Bi84 NS exhibits exceptional performance for CO2 reduction to formate, with the faradaic efficiency (FE) approaching â¼100% and a corresponding partial current density of 14.1 mA cm-2 at -0.94 V [vs the reversible hydrogen electrode (RHE)]. Furthermore, the FE could be maintained above 90% in a wide potential window (-0.84 to -1.54 V vs the RHE). This superior performance is attributed to the tuned electronic properties induced by the synergistic interaction between In and Bi, enabling the intermediates to be stably adsorbed on the catalyst surface to generate more formate ions.
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Background: Skin microbiota is important for maintenance of skin homeostasis; however, its disturbance may cause an increase in pathogenic microorganisms. Therefore, we aimed to develop a red ginseng formulation that can selectively promote beneficial bacteria. Methods: The effects of red ginseng formulation on microorganism growth were analyzed by comparing the growth rates of Staphylococcus aureus, S. epidermidis, and Cutibacterium acnes. Various preservatives mixed with red ginseng formulation were evaluated to determine the ideal composition for selective growth promotion of S. epidermidis. Red ginseng formulation with selected preservative was loaded into a biocompatible polymer mixture and applied to the faces of 20 female subjects in the clinical trial to observe changes in the skin microbiome. Results: Red ginseng formulation promoted the growth of S. aureus and S. epidermidis compared to fructooligosaccharide. When 1,2-hexanediol was applied with red ginseng formulation, only S. epidermidis showed selective growth. The analysis of the release rates of ginsenoside-Rg1 and -Re revealed that the exact content of Pluronic F-127 was around 11%. The application of hydrogel resulted in a decrease in C. acnes in all subjects. In subjects with low levels of S. epidermidis, the distribution of S. epidermidis was significantly increased with the application of hydrogel formulation and total microbial species of subjects decreased by 50% during the clinical trial. Conclusion: We confirmed that red ginseng formulation with 1,2-hexanediol can help maintain skin homeostasis through improvement of skin microbiome.
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Techniques such as direct sequencing and PCR-RFLP (restriction fragment length polymorphism) are widely used to analyze the genotypes of livestock. However, these conventional methods have the disadvantage of taking a lot of time and incurring considerable cost. The allele-specific PCR method performs PCR using two primers, and a single nucleotide polymorphism (SNP) genotype can be identified through electrophoresis, saving time and cost. Highly accurate results can be obtained by designing specific primers according to the allele of the SNP under study, utilizing primer binding to a complementary matching sequence. In this study, we established a genotyping system with the AS-PCR technique, using SNPs related to the improvement of the meat quality and meat mass of Korean cattle. Using the PRIMER1 program, we designed specific primers for SNPs located at the 3 ' end, with one SNP marker in the HSPB1 gene related to meat quantity and two SNP markers in the ADH1C and FASN genes related to meat quality in cattle. AS-PCR was performed on 10 Korean cattle using the primers designed with this system, and the genotypes could be identified by the size of the PCR product amplified as a result of electrophoresis. In the case of the HSPB1 g.2352T⯠> â¯C SNP, the T allele was amplified to 148â¯bp, and the C allele was amplified to 222â¯bp. The ADH1C c.-64T⯠> â¯C SNP was amplified to 492â¯bp at the T allele and 330â¯bp at the C allele. The FASN g.17924G⯠> â¯A SNP A allele was amplified to 377â¯bp and the G allele to 507â¯bp. The results for each SNP genotype were verified using direct sequencing, which showed that the genotypes identified by direct sequencing and the genotypes identified by the AS-PCR method matched exactly. The AS-PCR method therefore appears to be valuable for use in a genotyping system.
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Various neurological and musculoskeletal disorders can induce pathologic toe walking and lead to changes in foot kinematics. In this study, we analyzed the differences in foot kinematics between toe walking and heel-toe walking (HW) in able-bodied individuals. Twenty young healthy adults performed three gaits: HW, comfortable-height toe walking (CTW), and maximum-height toe walking (MTW). Oxford Foot Model was used for gait analysis. Toe walking showed increase of forefoot plantarflexion and hindfoot internal rotation compared to HW. Thus, our results may help distinguish the pathologic mechanism of the equinus gait in various disorders from the kinematic change of toe walking itself.
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Pé , Marcha , Adulto , Fenômenos Biomecânicos , Calcanhar , Humanos , Dedos do Pé , CaminhadaRESUMO
This case report presents a rare case of cerebral venous thrombosis (CVT) caused by spontaneous intracranial hypotension (SIH). The cause and prognosis of CVT can vary; CVT caused by SIH is uncommon and difficult to diagnose and treat. In this case, magnetic resonance imaging myelography showed definite cerebrospinal fluid leakage, and the patient's symptoms did not improve after conventional treatment. Furthermore, subdural hematoma occurred, causing mental deterioration; however, it improved dramatically after the blood patch procedure and burr hole drainage, which was performed after early cessation of anticoagulant therapy.
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How does vaccination against foot-and-mouth disease (FMD) affect pregnant cows? Vaccination is the most effective method of preventing the spread of FMD, but it is linked to sporadic side effects, such as abortion and premature birth, which result in economic loss. In this study, ruminoreticular temperature and body activity were measured before and after FMD vaccination using a ruminoreticular biocapsule sensor in Hanwoo cows at different stages of pregnancy. Compared to the unvaccinated groups, the ruminoreticular temperature increased 12 h after vaccination in the vaccinated groups. This increase in temperature is significantly correlated to vaccination. Compared to the nonpregnant and early pregnancy groups, the ruminoreticular temperature of the late pregnancy group increased sharply by more than 40 °C. Moreover, in nonpregnant and early pregnancy groups, a rapid increase in body activity was observed after FMD vaccinations. Of the 73 pregnant vaccinated cows in the study, a total of five cases had side effects (four abortions and one premature birth). Therefore, changes in the ruminoreticular temperature and activity in pregnant cows can be used as raw data to further clarify the association of FMD vaccination with the loss of a fetus and possibly predict abortion, miscarriage, and premature birth following FMD vaccination.
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Solar-driven reactive oxygen species (ROS) generation is an attractive disinfection technique for cell death and water purification. However, most photocatalysts require high stability in the water environment and the production of ROS with a sufficient amount and diffusion length to damage pathogens. Here, a ROS generation system was developed consisting of tapered crystalline silicon microwires coated with anatase titanium dioxide for a conformal junction. The system effectively absorbed >95% of sunlight over 300-1100 nm, resulting in effective ROS generation. The system was designed to produce various ROS species, but a logistic regression analysis with cellular survival data revealed that the diffusion length of the ROS is â¼9 µm, implying that the most dominant species causing cell damage is H2O2. Surprisingly, a quantitative analysis showed that only 15 min of light irradiation on the system would catalyze a local bactericidal effect comparable to the conventional germicidal level of H2O2 (â¼3 mM).
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Peróxido de Hidrogênio , Luz Solar , Morte Celular , Espécies Reativas de Oxigênio , TitânioRESUMO
Complex transportation systems often produce combined exposure to aircraft and road noise. Depending on the noise source, the annoyance response is different, and a masking effect occurs between the noise sources within the combined noise. Considering these characteristics, partial loudness was adopted to evaluate noise annoyance. First, a partial loudness model incorporating binaural inhibition was proposed and validated. Second, short- and long-term annoyance models were developed using partial loudness. Finally, the annoyance of combined noise was visualized as a map. These models can evaluate the annoyance by considering both the intensity and frequency characteristics of the noise. In addition, it is possible to quantify the masking effect that occurs between noise sources. Combined noise annoyance maps depict the degree of annoyance of residents and show the background noise effect, which is not seen on general noise maps.