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Designing plasmonic nanoparticles for biomedical photoacoustic (PA) imaging involves tailoring material properties at the nanometer scale. A key in developing plasmonic PA contrast nanoagents is to engineer their enhanced optical responses in the near-infrared wavelength range, as well as heat transfer properties and photostability. This study introduces anisotropic plasmonic nanosphere aggregates with close interparticle proximity as photostable and efficient contrast agent for PA imaging. Silver (Ag), among plasmonic metals, is particularly attractive due to its strongest optical response and highest heat conductivity. Our results demonstrate that close interparticle proximity in silver nanoaggregates (AgNAs), spatially confined within a polymer shell layer, leads to blackbody-like optical absorption, resulting in robust PA signals through efficient pulsed heat generation and transfer. Additionally, our AgNAs exhibit a high photodamage threshold highlighting their potential to outperform conventional plasmonic contrast agents for high-contrast PA imaging over multiple imaging sessions. Furthermore, we demonstrate the capability of the AgNAs for molecular PA cancer imaging in vivo by incorporating a tumor-targeting peptide moiety.
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Structural parameters play a crucial role in determining the electromagnetic and thermal responses of gold nanoconstructs (GNCs) at near-infrared (NIR) wavelengths. Therefore, developing GNCs for reliable, high-contrast photoacoustic imaging has been focused on adjusting structural parameters to achieve robust NIR light absorption with photostability. In this study, we introduce an efficient photoacoustic imaging contrast agent: gold sphere chains (GSCs) consisting of plasmonically coupled gold nanospheres. The chain geometry results in enhanced photoacoustic signal generation originating from outstanding photothermal characteristics compared to traditional gold contrast agents, such as gold nanorods. Furthermore, the GSCs produce consistent photoacoustic signals at laser fluences within the limits set by the American National Standards Institute. The exceptional photoacoustic response of GSCs allows for high-contrast photoacoustic imaging over multiple imaging sessions. Finally, we demonstrate the utility of our GSCs for molecular photoacoustic cancer imaging, both in vitro and in vivo, through the integration of a tumor-targeting moiety.
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OBJECTIVE: The aim of the present study was to explore inequalities in antenatal care (ANC) in South Korea. Based on the guidelines of the WHO, we categorized less than eight visits to an obstetrical facility as insufficient ANC. We examined ANC inequalities associated with age, disability, nationality, income, and geographic accessibility. METHODS: We extracted delivery event claimed from 2013 to 2022 from the National Health Insurance Service database. By tracing back 270 days from the delivery date, we counted the number of antenatal visits for each childbirth and calculated the proportion of women with insufficient ANC and assessed both absolute and relative inequalities for each population group. The logistic regression analysis for both underuse and overuse of ANC were conducted. RESULTS: Out of 3 416 517 childbirths, 104 109 women (3.0%) had fewer than eight ANC visits. Although the average number of ANC visits reached 18.7 in 2022, significant inequalities persisted across different population groups. The insufficient ANC rate was 28.1% for teenage women, 6.4% for disabled women, 10.7% for non-Korean women, and 15.2% for dependents of medical aid households. Women with low income and those living in obstetric care underserved areas also exhibited higher ANC insufficiency. From 2021 to 2022, sufficiency in ANC decreased for teenage, disabled, and non-Korean women, highlighting the effects of the COVID-19 pandemic. CONCLUSION: Antenatal care inequalities are evident in South Korea's well-resourced health system. There is a need for further investigation into these disparities and the qualitative aspects of maternity care services.
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Disparidades em Assistência à Saúde , Cuidado Pré-Natal , Humanos , Feminino , República da Coreia , Cuidado Pré-Natal/estatística & dados numéricos , Gravidez , Adulto , Disparidades em Assistência à Saúde/estatística & dados numéricos , Adulto Jovem , Adolescente , Acessibilidade aos Serviços de Saúde/estatística & dados numéricos , Fatores Socioeconômicos , Pessoa de Meia-IdadeRESUMO
In 2019, a child's death in Korea led to legislation that imposed stricter penalties for school zone traffic violations. We assessed the impact of that legislation using 2017-2022 Traffic Accident Analysis System data. Adjusted analyses revealed a significant decline in severe injuries in school zones, decreasing from 11 cases to 8 cases per month (p=0.017). The legislation correlated with a reduced risk of all child traffic injuries (risk ratio, 0.987; 95% confidence interval, 0.977 to 0.997; p=0.002), indicating its efficacy in curbing accidents.
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Acidentes de Trânsito , Análise de Séries Temporais Interrompida , Instituições Acadêmicas , Ferimentos e Lesões , Humanos , República da Coreia/epidemiologia , Acidentes de Trânsito/prevenção & controle , Criança , Ferimentos e Lesões/prevenção & controle , Ferimentos e Lesões/epidemiologia , Punição , Crime/prevenção & controle , Masculino , Feminino , AdolescenteRESUMO
The global threat posed by the COVID-19 pandemic has catalyzed the development of point-of-care (POC) molecular diagnostics. While loop-mediated isothermal amplification (LAMP) stands out as a promising technique among FDA-approved methods, it is occasionally susceptible to a high risk of false positives due to nonspecific amplification of a primer dimer. In this work, we report an enhancing LAMP technique in terms of assay sensitivity and reliability through streamlined integration with a nonpowered nanoelectric preconcentration (NPP). The NPP, serving as a sample preparation tool, enriched the virus concentration in samples prior to the subsequent LAMP assay. This enrichment enabled not only to achieve more sensitive assay but also to shorten the assay time for all tested clinical samples by â¼10 min compared to the conventional LAMP. The shortened assay time suppresses the occurrence of nonspecific amplification by not providing the necessary incubation time, effectively suppressing misidentification by false positives. Utilizing this technique, we also developed a prototype of the POC NPP-LAMP kit. This kit offers a streamlined diagnostic process for nontrained individuals, from the sample enrichment, transfer of the enriched sample to LAMP assays, which facilitates on-site/on-demand diagnosis of SARS-CoV-2. This development holds the potential to contribute toward preventing not only the current outbreak but also future occurrences of pandemic viruses.
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COVID-19 , Pandemias , Humanos , Reprodutibilidade dos Testes , Técnicas de Diagnóstico Molecular/métodos , COVID-19/diagnóstico , Técnicas de Amplificação de Ácido Nucleico/métodos , Sensibilidade e Especificidade , RNA ViralRESUMO
In plasmonic nanoconstructs (NCs), fine-tuning interparticle interactions at the subnanoscale offer enhanced electromagnetic and thermal responses in the near-infrared (NIR) wavelength range. Due to tunable electromagnetic and thermal characteristics, NCs can be excellent photoacoustic (PA) imaging contrast agents. However, engineering plasmonic NCs that maximize light absorption efficiency across multiple polarization directions, i.e., exhibiting blackbody absorption behavior, remains challenging. Herein, we present the synthesis, computational simulation, and characterization of hyper-branched gold nanoconstructs (HBGNCs) as a highly efficient PA contrast agent. HBGNCs exhibit remarkable optical properties, including strong NIR absorption, high absorption efficiency across various polarization angles, and superior photostability compared to conventional standard plasmonic NC-based contrast agents such as gold nanorods and gold nanostars. In vitro and in vivo experiments confirm the suitability of HBGNCs for cancer imaging, showcasing their potential as reliable PA contrast agents and addressing the need for enhanced imaging contrast and stability in bioimaging applications.
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Neoplasias , Técnicas Fotoacústicas , Humanos , Ouro , Técnicas Fotoacústicas/métodos , Meios de Contraste , Diagnóstico por Imagem/métodosRESUMO
Metabolic reprogramming has been recognized as one of the major mechanisms that fuel tumor initiation and progression. Our previous studies demonstrate that activation of Drp1 promotes fatty acid oxidation and downstream Wnt signaling. Here we investigate the role of Drp1 in regulating glycogen metabolism in colon cancer. Knockdown of Drp1 decreases mitochondrial respiration without increasing glycolysis. Analysis of cellular metabolites reveals that the levels of glucose-6-phosphate, a precursor for glycogenesis, are significantly elevated whereas pyruvate and other TCA cycle metabolites remain unchanged in Drp1 knockdown cells. Additionally, silencing Drp1 activates AMPK to stimulate the expression glycogen synthase 1 (GYS1) mRNA and promote glycogen storage. Using 3D organoids from Apcf/f/Villin-CreERT2 models, we show that glycogen levels are elevated in tumor organoids upon genetic deletion of Drp1. Similarly, increased GYS1 expression and glycogen accumulation are detected in xenograft tumors derived from Drp1 knockdown colon cancer cells. Functionally, increased glycogen storage provides survival advantage to Drp1 knockdown cells. Co-targeting glycogen phosphorylase-mediated glycogenolysis sensitizes Drp1 knockdown cells to chemotherapy drug treatment. Taken together, our results suggest that Drp1-loss activates glucose uptake and glycogenesis as compensative metabolic pathways to promote cell survival. Combined inhibition of glycogen metabolism may enhance the efficacy of chemotherapeutic agents for colon cancer treatment.
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Neoplasias do Colo , Glicogenólise , Humanos , Sobrevivência Celular , Dinâmica Mitocondrial , Transformação Celular Neoplásica , Glicogênio/metabolismo , Neoplasias do Colo/genética , Dinaminas/metabolismoRESUMO
Stem cell therapy has immense potential in a variety of regenerative medicine applications. However, clinical stem cell therapy is severely limited by challenges in assessing the location and functional status of implanted cells in vivo. Thus, there is a great need for longitudinal, noninvasive stem cell monitoring. Here we introduce a multidisciplinary approach combining nanosensor-augmented stem cell labeling with ultrasound guided photoacoustic (US/PA) imaging for the spatial tracking and functional assessment of transplanted stem cell fate. Specifically, our nanosensor incorporates a peptide sequence that is selectively cleaved by caspase-3, the primary effector enzyme in mammalian cell apoptosis; this cleavage event causes labeled cells to show enhanced optical absorption in the first near-infrared (NIR) window. Optimization of labeling protocols and spectral characterization of the nanosensor in vitro showed a 2.4-fold increase in PA signal from labeled cells during apoptosis while simultaneously permitting cell localization. We then successfully tracked the location and apoptotic status of mesenchymal stem cells in a mouse hindlimb ischemia model for 2 weeks in vivo, demonstrating a 4.8-fold increase in PA signal and spectral slope changes in the first NIR window under proapoptotic (ischemic) conditions. We conclude that our nanosensor allows longitudinal, noninvasive, and nonionizing monitoring of stem cell location and apoptosis, which is a significant improvement over current end-point monitoring methods such as biopsies and histological staining of excised tissue.
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Células-Tronco Mesenquimais , Técnicas Fotoacústicas , Camundongos , Animais , Caspase 3 , Transplante de Células-Tronco , Apoptose , Técnicas Fotoacústicas/métodos , MamíferosRESUMO
A simple, affordable point of care test (POCT) is necessary for on-site detection of coronavirus disease 2019 (COVID-19). The lateral flow assay (LFA) has great potential for use in POCT mainly because of factors such as low time consumption, low cost, and ease of use. However, it lacks sensitivity and limits of detection (LOD), which are essential for early diagnostics. In this study, we proposed a non-powered preconcentrator (NPP) based on nanoelectrokinetics for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) Antigen (Ag) lateral flow assay. The non-powered preconcentrator is composed of glass fiber-based composite paper and ion permselective material, and it can be simply operated by force balancing gravitational, capillary, and depletion-induced forces. The proposed approach helps enrich the SARS-CoV-2 viral nucleocapsid (N) proteins based on a 10-min operation, and it improved the LOD by up to 10-fold. The corresponding virus enrichment, which was evaluated using the reverse-transcriptase polymerase chain reaction (RT-PCR), revealed an improvement in ΔCt values > 3. We successfully demonstrated the enhancement of the NPP-assisted LFA, we extended to applying it to clinical samples. Further, we demonstrated an affordable, easy-to-implement form of LFA by simply designing NPP directly on the LFA buffer tube.
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Técnicas Biossensoriais , COVID-19 , Humanos , COVID-19/diagnóstico , SARS-CoV-2 , Teste para COVID-19 , Limite de Detecção , Sensibilidade e EspecificidadeRESUMO
The number of smartwatch users has been rapidly increasing in recent years. A smartwatch is a wearable device that collects various types of data using sensors and provides basic functions, such as healthcare-related measurements and audio recording. In this study, we proposed the forensic authentication method for audio recordings from the Voice Recording application in the Samsung Galaxy Watch4 series. First, a total of 240 audio recordings from each of the four different models, paired with four different smartphones for synchronization via Bluetooth, were collected and verified. To analyze the characteristics of smartwatch audio recordings, we examined the transition of the audio latency, writable audio bandwidth, timestamps, and file structure between those generated in the smartwatches and those edited using the Voice Recording application of the paired smartphones. In addition, the devices with the audio recordings were examined via the Android Debug Bridge (ADB) tool and compared with the timestamps stored in the file system. The experimental results showed that the audio latency, writable audio bandwidth, and file structure of audio recordings generated by smartwatches differed from those generated by smartphones. Additionally, by analyzing the file structure, audio recordings can be classified as unmanipulated, manipulation has been attempted, or manipulated. Finally, we can forensically authenticate the audio recordings generated by the Voice Recorder application in the Samsung Galaxy Watch4 series by accessing the smartwatches and analyzing the timestamps related to the audio recordings in the file system.
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Gravação de Som , Dispositivos Eletrônicos Vestíveis , Smartphone , Medicina LegalRESUMO
Manipulating matter at the nanometer scale to create desired plasmonic nanostructures holds great promise in the field of biomedical photoacoustic (PA) imaging. We demonstrate a strategy for regulating PA signal generation from anisotropic nano-sized assemblies of gold nanospheres (Au NSs) by adjusting the inter-particle connectivity between neighboring Au NSs. The inter-particle connectivity is controlled by modulating the diameter and inter-particle spacing of Au NSs in the nanoassemblies. The results indicate that nanoassemblies with semi-connectivity, i.e., assemblies with a finite inter-particle spacing shorter than the theoretical limit of repulsion between nearby Au NSs, exhibit 3.4-fold and 2.4-fold higher PA signals compared to nanoassemblies with no connectivity and full connectivity, respectively. Furthermore, due to the reduced diffusion of Au atoms, the semi-connectivity Au nanoassemblies demonstrate high photodamage threshold and, therefore, excellent photostability at fluences above the current American National Standards Institute limits. The exceptional photostability of the semi-connectivity nanoassemblies highlights their potential to surpass conventional plasmonic contrast agents for continuing PA imaging. Collectively, our findings indicate that semi-connected nanostructures are a promising option for reliable, high-contrast PA imaging applications over multiple imaging sessions due to their strong PA signals and enhanced photostability.
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The mammalian STE 20-like protein kinase 4 (MST4) gene is highly expressed in several cancer types, but little is known about the role of MST4 in breast cancer, and the function of MST4 during epithelial-mesenchymal transition (EMT) has not been fully elucidated. Here we report that overexpression of MST4 in breast cancer results in enhanced cell growth, migration, and invasion, whereas inhibition of MST4 expression significantly attenuates these properties. Further study shows that MST4 promotes EMT by activating Akt and its downstream signaling molecules such as E-cadherin/N-cadherin, Snail, and Slug. MST4 also activates AKT and its downstream pro-survival pathway. Furthermore, by analyzing breast cancer patient tissue microarray and silicon datasets, we found that MST4 expression is much higher in breast tumor tissue compared to normal tissue, and significantly correlates with cancer stage, lymph node metastasis and a poor overall survival rate (p < 0.05). Taken together, our findings demonstrate the oncogenic potential of MST4 in breast cancer, highlighting its role in cancer cell proliferation, migration/invasion, survival, and EMT, suggesting a possibility that MST4 may serve as a novel therapeutic target for breast cancer.
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Neoplasias da Mama , Proteínas Serina-Treonina Quinases , Proteínas Proto-Oncogênicas c-akt , Feminino , Humanos , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/genética , Neoplasias da Mama/metabolismo , Transição Epitelial-Mesenquimal/genética , Metástase Linfática , Oncogenes , Proteínas Proto-Oncogênicas c-akt/metabolismo , Transdução de Sinais , Proteínas Serina-Treonina Quinases/genéticaRESUMO
Acute myeloid leukemia (AML) is an aggressive blood cancer with a high rate of relapse associated with adverse survival outcomes, especially in elderly patients. An aberrant expression of cyclin dependent kinase 7 (CDK7) is associated with poor outcomes and CDK7 inhibition has showed antitumor activities in various cancers. We investigated the efficacy of YPN-005, a CDK7 inhibitor in AML cell lines, xenograft mouse model, and primary AML cells. YPN-005 effectively inhibited the proliferation of AML cells by inducing apoptosis and reducing phosphorylation of RNA polymerase II. The c-MYC expression decreased with treatment of YPN-005, and the effect of YPN-005 was negatively correlated with c-MYC expression. YPN-005 also showed antileukemic activities in primary AML cells, especially those harboring FMS-like tyrosine kinase 3-internal tandem duplication (FLT3-ITD) mutation and in in vivo mouse model. Phosphorylated FLT3/Signal transducer and activator of transcription 5 (STAT5) was decreased and FLT3/STAT5 was downregulated with YPN-005 treatment. Our data suggest that YPN-005 has a role in treating AML by suppressing c-MYC and FLT3.
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OBJECTIVES: This study analyzed inequities in excess mortality according to region and socioeconomic position to explain the distribution of excess mortality in Korea in 2020. METHODS: We acquired weekly all-cause mortality data from January 2015 to December 2020 from (1) the National Health Insurance Database and (2) Vital Statistics. Excess mortality for 2020 was calculated by comparing the weekly observed and expected deaths from the same period (2015-2019) using quasi-Poisson regression. RESULTS: An inequitable distribution of excess mortality was identified. The estimated excess mortality in Korea was -29,112 (95% confidence interval, -29,832 to -28,391), corresponding to -55 per 100,000, and the ratio of observed deaths to expected deaths was 0.91. Negative excess mortality was observed except for females in the 0-14 age group. Male Medical Aid beneficiaries showed positive excess mortality, while non-disabled and disabled groups showed similar negative values. When the standardized mortality ratio was calculated for the top 10 causes of death, deaths from Alzheimer's disease and septicemia increased, whereas those from diabetes mellitus and cerebrovascular disease decreased. The decrease in mortality was primarily concentrated in older adults, while the mortality of young females increased due to increased intentional self-harm. CONCLUSIONS: This study adds essential evidence regarding the overall performance of Korea. The observed inequalities according to various socioeconomic variables indicate that the results of strict measures to control coronavirus disease 2019 were not distributed equitably. Efforts should be made to properly evaluate the current and future problems related to the pandemic.
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COVID-19 , Feminino , Humanos , Masculino , Idoso , Pandemias , República da Coreia/epidemiologia , MortalidadeRESUMO
Objective: This study assessed the relationship between self-rated political orientation and attitude toward the cash transfer policy during the COVID-19 pandemic. Methods: This cross-sectional study conducted in South Korea during the pandemic included a stratified sample of 1,004 respondents (aged 19 years and older). We tested the hypotheses that political orientation shapes attitudes toward social policies and that other socioeconomic factors might have relatively minor importance. Logistic regression was used to identify associations between political orientation and attitude toward the cash transfer policy. Average marginal effects were calculated to determine the effect size of each variable. Results: Political orientation, age, and residential area were correlated with attitudes toward the policy. Compared to the conservatives, the non-committed and the moderate showed about 10% more favorable attitudes, and the progressive group showed robust support. People in their 30s and 40s showed similar attitudes to the 18-29 group, while older people showed much lower support. Compared to the Seoul metropolitan area, residents of the Ho-nam area showed favorable attitudes, and those of the Yeong-nam area had relatively unfavorable attitudes. Conclusions: This study suggests that attitudes toward the cash transfer policy are mainly associated with political orientation. Although these results illuminate pandemics' social and political dimensions, further efforts are needed to fully understand the determinants and mechanisms of attitudes toward policies outside the traditional health policy scope.
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COVID-19 , Pandemias , Idoso , Atitude , COVID-19/epidemiologia , Estudos Transversais , Humanos , Política Pública , República da Coreia/epidemiologiaRESUMO
A lateral flow assay (LFA) platform is a powerful tool for point-of-care testing (POCT), especially for self-testing. Although the LFA platform provides a simple and disposable tool for Coronavirus disease of 2019 (COVID-19) antigen (Ag) and antibody (Ab) screening tests, the lower sensitivity for low virus titers has been a bottleneck for practical applications. Herein, we report the combination of a microfluidic paper-based nanoelectrokinetic (NEK) preconcentrator and an LFA platform for enhancing the sensitivity and limit of detection (LOD). Biomarkers were electrokinetically preconcentrated onto a specific layer using the NEK preconcentrator, which was then coupled with LFA diagnostic devices for enhanced performance. Using this nanoelectrokinetic-assisted LFA (NEK-LFA) platform for self-testing, the severe acute respiratory syndrome coronavirus 2 Immunoglobulin G (SARS-CoV-2 IgG) sample was preconcentrated from serum samples. After preconcentration, the LOD of the LFA was enhanced by 32-fold, with an increase in analytical sensitivity (16.4%), which may offer a new opportunity for POCT and self-testing, especially in the COVID-19 pandemic and endemic global context.
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Técnicas Biossensoriais , COVID-19 , Anticorpos Antivirais , COVID-19/diagnóstico , Humanos , Imunoensaio , Pandemias , SARS-CoV-2 , Sensibilidade e EspecificidadeRESUMO
Multivalent O-antigen polysaccharide glycoconjugate vaccines are under development to prevent invasive infections caused by pathogenic Enterobacteriaceae. Sequence type 131 (ST131) Escherichia coli of serotype O25b has emerged as the predominant lineage causing invasive multidrug-resistant extraintestinal pathogenic E. coli (ExPEC) infections. We observed the prevalence of E. coli O25b ST131 among a contemporary collection of isolates from U.S. bloodstream infections from 2013 to 2016 (n = 444) and global urinary tract infections from 2014 to 2017 (n = 102) to be 25% and 24%, respectively. To maximize immunogenicity of the serotype O25b O antigen, we investigated glycoconjugate properties, including CRM197 carrier protein cross-linking (single-end versus cross-linked "lattice") and conjugation chemistry (reductive amination chemistry in dimethyl sulfoxide [RAC/DMSO] versus ((2-((2-oxoethyl)thio)ethyl)carbamate [eTEC] linker). Using opsonophagocytic assays (OPAs) to measure serum functional antibody responses to vaccination, we observed that higher-molecular-mass O25b long-chain lattice conjugates showed improved immunogenicity in mice compared with long- or short-chain O antigens conjugated via single-end attachment. The lattice conjugates protected mice from lethal challenge with acapsular O25b ST131 strains as well as against hypervirulent O25b isolates expressing K5 or K100 capsular polysaccharides. A single 1-µg dose of long-chain O25b lattice conjugate constructed with both chemistries also elicited robust serum IgG and OPA responses in cynomolgus macaques. Our findings show that key properties of the O-antigen carrier protein conjugate such as saccharide epitope density and degree of intermolecular cross-linking can significantly enhance functional immunogenicity.
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Infecções por Escherichia coli , Antígenos O , Animais , Proteínas de Transporte , Escherichia coli , Infecções por Escherichia coli/prevenção & controle , Glicoconjugados , CamundongosRESUMO
In this study, we propose an advanced forensic examination procedure for audio recordings generated by the Voice Memos application with iPhone Operation System (iOS)14, to verify that these are the original recordings and have not been manipulated. The proposed examination procedure consists of an analysis of the characteristics of audio recordings and of the file system of the device storing the audio recordings. To analyze the characteristics of audio recordings, we compare the encoding parameters (bitrate, sampling rate, timestamps, etc.) and the file structure to determine whether audio recordings were manipulated. Next, in the device examination step, we analyze the media-log history and temporary files of the file system obtained by mobile forensic tools. For comparative analysis, a total of 100 audio recording samples were obtained through the Voice Memos application from five iPhone mobile handsets of different models with iOS14 installed using Advanced audio coding (AAC) or Apple lossless audio codec (ALAC). As a result of analyzing the encoding parameters between the original and manipulated audio recordings, as well as the temporary files contained in the device file system, the difference in the encoding parameters and the very unique trace of the original audio recordings in the temporary files were confirmed when manipulating the audio recordings. In particular, the primary advantage of our proposed method is its potential ability to recover original audio recordings that were subsequently manipulated via the temporary files examined in the device file system analysis.
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Matrix metalloproteinase-9 (MMP-9) plays major roles in extracellular matrix (ECM) remodeling and membrane protein cleavage, suggesting a high correlation with cancer cell invasion and tumor metastasis. Here, we present a contrast agent based on a DNA aptamer that can selectively target human MMP-9 in the tumor microenvironment (TME) with high affinity and sensitivity. Surface modification of plasmonic gold nanospheres with the MMP-9 aptamer and its complementary sequences allows the nanospheres to aggregate in the presence of human MMP-9 through DNA displacement and hybridization. Aggregation of gold nanospheres enhances the optical absorption in the first near-infrared window (NIR-I) due to the plasmon coupling effect, thereby allowing us to detect the aggregated gold nanospheres within the TME via ultrasound-guided photoacoustic (US/PA) imaging. Selective and sensitive detection of human MMP-9 via US/PA imaging is demonstrated in solution of nanosensors with the pre-treatment of human MMP-9, in vitro in cell culture, and in vivo in a xenograft murine model of human breast cancer.
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The disturbance of strictly regulated self-regeneration in mammalian intestinal epithelium is associated with various intestinal disorders, particularly inflammatory bowel diseases (IBDs). TNFα, which plays a critical role in the pathogenesis of IBDs, has been reported to inhibit production of ketone bodies such as ß-hydroxybutyrate (ßHB). However, the role of ketogenesis in the TNFα-mediated pathological process is not entirely known. Here, we showed the regulation and role of HMGCS2, the rate-limiting enzyme of ketogenesis, in TNFα-induced apoptotic and inflammatory responses in intestinal epithelial cells. Treatment with TNFα dose-dependently decreased protein and mRNA expression of HMGCS2 and its product, ßHB production in human colon cancer cell lines HT29 and Caco2 cells and mouse small intestinal organoids. Moreover, the repressed level of HMGCS2 protein was found in intestinal epithelium of IBD patients with Crohn's disease and ulcerative colitis as compared with normal tissues. Furthermore, knockdown of HMGCS2 enhanced and in contrast, HMGCS2 overexpression attenuated, the TNFα-induced apoptosis and expression of pro-inflammatory chemokines (CXCL1-3) in HT29, Caco2 cells and DLD1 cells, respectively. Treatment with ßHB or rosiglitazone, an agonist of PPARγ, which increases ketogenesis, attenuated TNFα-induced apoptosis in the intestinal epithelial cells. Finally, HMGCS2 knockdown enhanced TNFα-induced reactive oxygen species (ROS) generation. In addition, hydrogen peroxide, the major ROS contributing to intestine injury, decreased HMGCS2 expression and ßHB production in the intestinal cells and mouse organoids. Our findings demonstrate that increased ketogenesis attenuates TNFα-induced apoptosis and inflammation in intestinal cells, suggesting a protective role for ketogenesis in TNFα-induced intestinal pathologies.