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1.
Vaccines (Basel) ; 12(5)2024 Apr 29.
Artigo em Inglês | MEDLINE | ID: mdl-38793727

RESUMO

Outbreaks caused by foot-and-mouth disease (FMD) A/ASIA/G-VII lineage viruses have often occurred in Middle Eastern and Southeast Asian countries since 2015. Because A/ASIA/G-VII lineage viruses are reported to have distinct antigenic relatedness with available commercial FMD vaccine strains, it is necessary to investigate whether inoculation with vaccines used in Korea could confer cross-protection against A/ASIA/G-VII lineage viruses. In the present study, we conducted two vaccination challenge trials to evaluate the efficacy of three commercial FMD vaccines (O/Manisa + O/3039 + A/Iraq, O/Campos + A/Cruzeiro + A/2001, and O/Primorsky + A/Zabaikalsky) against heterologous challenge with ASIA/G-VII lineage viruses (A/TUR/13/2017 or A/BHU/3/2017 strains) in pigs. In each trial, clinical signs, viremia, and salivary shedding of virus were measured for 7 days after challenge. In summary, the O/Campos + A/Cruzeiro + A/2001 vaccine provided full protection against two A/ASIA/G-VII lineage viruses in vaccinated pigs, where significant protection was observed. Although unprotected animals were observed in groups vaccinated with O/Manisa + O/3039 + A/Iraq or O/Primorsky + A/Zabaikalsky vaccines, the clinical scores and viral RNA levels in the sera and oral swabs of vaccinated animals were significantly lower than those of unvaccinated controls.

2.
Artigo em Inglês | MEDLINE | ID: mdl-37632203

RESUMO

Rare but consistent reports of abscopal remission in patients challenge the notion that radiotherapy (RT) is a local treatment; radiation-induced cancer cell death can trigger activation and recruitment of dendritic cells to the primary tumor site, which subsequently initiates systemic immune responses against metastatic lesions. Although this abscopal effect was initially considered an anomaly, combining RT with immune checkpoint inhibitor therapies has been shown to greatly improve the incidence of abscopal responses via modulation of the immunosuppressive tumor microenvironment. Preclinical studies have demonstrated that nanomaterials can further improve the reliability and potency of the abscopal effect for various different types of cancer by (1) altering the cell death process to be more immunogenic, (2) facilitating the capture and transfer of tumor antigens from the site of cancer cell death to antigen-presenting cells, and (3) co-delivering immune checkpoint inhibitors along with radio-enhancing agents. Several unanswered questions remain concerning the exact mechanisms of action for nanomaterial-enhanced RT and for its combination with immune checkpoint inhibition and other immunostimulatory treatments in clinically relevant settings. The purpose of this article is to summarize key recent developments in this field and also highlight knowledge gaps that exist in this field. An improved mechanistic understanding will be critical for clinical translation of nanomaterials for advanced radio-immunotherapy. This article is categorized under: Therapeutic Approaches and Drug Discovery > Nanomedicine for Oncologic Disease.


Assuntos
Nanoestruturas , Neoplasias , Humanos , Reprodutibilidade dos Testes , Imunoterapia , Neoplasias/radioterapia , Nanotecnologia , Nanoestruturas/uso terapêutico , Microambiente Tumoral
3.
Molecules ; 28(16)2023 Aug 16.
Artigo em Inglês | MEDLINE | ID: mdl-37630339

RESUMO

The issue of food fraud has become a significant global concern as it affects both the quality and safety of food products, ultimately resulting in the loss of customer trust and brand loyalty. To address this problem, we have developed an innovative approach that can tackle various types of food fraud, including adulteration, substitution, and dilution. Our methodology utilizes an integrated system that combines laser-induced breakdown spectroscopy (LIBS) and Raman spectroscopy. Although both techniques emerged as valuable tools for food analysis, they have until now been used separately, and their combined potential in food fraud has not been thoroughly tested. The aim of our study was to demonstrate the potential benefits of integrating Raman and LIBS modalities in a portable system for improved product classification and subsequent authentication. In pursuit of this objective, we designed and tested a compact, hybrid Raman/LIBS system, which exhibited distinct advantages over the individual modalities. Our findings illustrate that the combination of these two modalities can achieve higher accuracy in product classification, leading to more effective and reliable product authentication. Overall, our research highlights the potential of hybrid systems for practical applications in a variety of industries. The integration and design were mainly focused on the detection and characterization of both elemental and molecular elements in various food products. Two different sets of solid food samples (sixteen Alpine-style cheeses and seven brands of Arabica coffee beans) were chosen for the authentication analysis. Class detection and classification were accomplished through the use of multivariate feature selection and machine-learning procedures. The accuracy of classification was observed to improve by approximately 10% when utilizing the hybrid Raman/LIBS spectra, as opposed to the analysis of spectra from the individual methods. This clearly demonstrates that the hybrid system can significantly improve food authentication accuracy while maintaining the portability of the combined system. Thus, the successful implementation of a hybrid Raman-LIBS technique is expected to contribute to the development of novel portable devices for food authentication in food as well as other various industries.


Assuntos
Queijo , Análise Espectral Raman , Contaminação de Medicamentos , Fraude , Indústrias
4.
Biomater Sci ; 11(18): 6311-6324, 2023 Sep 12.
Artigo em Inglês | MEDLINE | ID: mdl-37552121

RESUMO

Photodynamic therapy (PDT) using 5-aminolevulinic acid (ALA) prodrug is a clinically tried and proven treatment modality for surface-level lesions. However, its use for deep-seated tumors has been limited due to the poor penetration depth of visible light needed to activate the photosensitizer protoporphyrin IX (PPIX), which is produced from ALA metabolism. Herein, we report the usage of poly(ethylene glycol-b-lactic acid) (PEG-PLA)-encapsulated calcium tungstate (CaWO4, CWO for short) nanoparticles (PEG-PLA/CWO NPs) as energy transducers for X-ray-activated PDT using ALA. Owing to the spectral overlap between radioluminescence afforded by the CWO core and the absorbance of PPIX, these NPs can serve as an in situ visible light activation source during radiotherapy (RT), thereby mitigating the limitation of penetration depth. We demonstrate that this effect is observed across different cell lines with varying radio-sensitivity. Importantly, both PPIX and PEG-PLA/CWO NPs exhibit no significant toxicities at therapeutic doses in the absence of radiation. To assess the efficacy of this approach, we conducted a study using a syngeneic mouse model subcutaneously implanted with inherently radio-resistant 4T1 tumors. The results show a significantly improved prognosis compared to conventional RT, even with as few as 2 fractions of 4 Gy X-rays. Taken together, these results suggest that PEG-PLA/CWO NPs are promising agents for application of ALA-PDT in deep-seated tumors, thereby significantly expanding the utility of the already established treatment strategy.


Assuntos
Nanopartículas , Neoplasias , Fotoquimioterapia , Pró-Fármacos , Animais , Camundongos , Ácido Aminolevulínico/farmacologia , Ácido Aminolevulínico/uso terapêutico , Pró-Fármacos/farmacologia , Pró-Fármacos/uso terapêutico , Fotoquimioterapia/métodos , Fármacos Fotossensibilizantes/farmacologia , Fármacos Fotossensibilizantes/uso terapêutico , Neoplasias/tratamento farmacológico , Nanopartículas/uso terapêutico , Linhagem Celular Tumoral
5.
Stem Cell Res Ther ; 14(1): 178, 2023 07 19.
Artigo em Inglês | MEDLINE | ID: mdl-37468918

RESUMO

BACKGROUND: Preclinical studies showed that mesenchymal stem cells (MSCs) ameliorate tau phosphorylation, amyloid-beta accumulation, and inflammation in Alzheimer's disease (AD) mouse models via secretion of neurotrophic factors and cytokines. We aimed to identify CSF biomarkers that can be used to predict or monitor the response to MSCs in patients with AD. METHODS: AD patients were injected with human umbilical cord blood-MSCs (n = 22) or placebo (n = 12). The cerebrospinal fluid (CSF) samples were collected at baseline, one day after the first injection, and one day after the third injection. The patients injected with MSCs were classified into good responder (GR) or poor responder (PR) groups based on the rate of changes in the ratio of total-tau and phosphorylated-tau in the CSF. We selected three typical participants in each group, and their CSF protein levels were analyzed using liquid chromatography/tandem mass spectrometry (LC-MS/MS). RESULTS: In the LC-MS/MS analysis, 1,667 proteins were identified. Eleven proteins showed significant differences between the typical GR and PR at baseline. Based on their significance level and known functions, two proteins, reticulocalbin-3 (RCN3) and follistatin-related protein 3 (FSTL3), were selected as potential biomarkers to predict MSC response. A total of 173 proteins showed significant change one day after the third injection compared to the baseline in typical GR. We excluded 45 proteins that showed significant change after the third injection compared to the baseline in the typical PR. Based on their significance level and known function, four proteins, scrapie-responsive protein 1 (SCRG1), neural proliferation differentiation and control protein (NPDC1), apolipoprotein E (ApoE), and cystatin C (CysC), were selected as potential biomarker to monitor MSC response. Additionally, functional analysis revealed that the increased CSF proteins after the third injection compared to the baseline in the typical GR were associated with synaptogenesis. CONCLUSIONS: This study identified two proteins (RCN3 and FSTL3) that may be potential biomarkers for predicting MSC response and four proteins (SCRG1, NPDC1, ApoE, CysC) that may be potential biomarkers for monitoring MSC response in patients with AD. Further studies are needed to validate our results. Trial registration Clinical Trials.gov, NCT02054208. Registered on 4 February 2014. Samsung Medical Center IRB File No.2017-04-025. Registered on 20 June 2017.


Assuntos
Doença de Alzheimer , Animais , Camundongos , Humanos , Doença de Alzheimer/terapia , Cromatografia Líquida , Proteínas tau/genética , Proteínas tau/metabolismo , Espectrometria de Massas em Tandem , Peptídeos beta-Amiloides , Apolipoproteínas E/metabolismo , Biomarcadores , Fragmentos de Peptídeos , Proteínas de Ligação ao Cálcio
6.
Mol Biotechnol ; 2023 Jul 10.
Artigo em Inglês | MEDLINE | ID: mdl-37428433

RESUMO

Potential threat of smallpox bioterrorism and concerns related to the adverse effects of currently licensed live-virus vaccines suggest the need to develop novel vaccines with better efficacy against smallpox. Use of DNA vaccines containing specific antigen-encoding plasmids prevents the risks associated with live-virus vaccines, offering a promising alternative to conventional smallpox vaccines. In this study, we investigated the efficiency of toll-like receptor (TLR) ligands in enhancing the immunogenicity of smallpox DNA vaccines. BALB/c mice were immunized with a DNA vaccine encoding the vaccinia virus L1R protein, along with the cytosine-phosphate-guanine (CpG) motif as a vaccine adjuvant, and their immune response was analyzed. Administration of B-type CpG oligodeoxynucleotides (ODNs) as TLR9 ligands 24 h after DNA vaccination enhanced the Th2-biased L1R-specific antibody immunity in mice. Moreover, B-type CpG ODNs improved the protective effects of the DNA vaccine against the lethal Orthopoxvirus challenge. Therefore, use of L1R DNA vaccines with CpG ODNs as adjuvants is a promising approach to achieve effective immunogenicity against smallpox infection.

7.
Int J Mol Sci ; 24(10)2023 May 12.
Artigo em Inglês | MEDLINE | ID: mdl-37239986

RESUMO

Let-7 miRNAs have pleiotropic cellular functions in cell proliferation, migration, and regenerative processes. Here, we investigate whether the inhibition of let-7 miRNAs with antisense oligonucleotides (ASOs) can be a transient and safe strategy enhancing the therapeutic potential of mesenchymal stromal cells (MSCs) to overcome their limitations in cell therapeutic trials. We first identified major subfamilies of let-7 miRNAs preferentially expressed in MSCs, and efficient ASO combinations against these selected subfamilies that mimic the effects of LIN28 activation. When let-7 miRNAs were inhibited with an ASO combination (anti-let7-ASOs), MSCs exhibited higher proliferation with delayed senescence during the passaging into a culture. They also exhibited increased migration and enhanced osteogenic differentiation potential. However, these changes in MSCs were not accompanied by cell-fate changes into pericytes or the additional acquisition of stemness, but instead occurred as functional changes accompanied by changes in proteomics. Interestingly, MSCs with let-7 inhibition exhibited metabolic reprogramming characterized by an enhanced glycolytic pathway, decreased reactive oxygen species, and lower transmembrane potential in mitochondria. Moreover, let-7-inhibited MSCs promoted the self-renewal of neighboring hematopoietic progenitor cells, and enhanced capillary formation in endothelial cells. These findings together show that our optimized ASO combination efficiently reprograms the MSC functional state, allowing for more efficient MSC cell therapy.


Assuntos
Células-Tronco Mesenquimais , MicroRNAs , Osteogênese , Oligonucleotídeos Antissenso/genética , Oligonucleotídeos Antissenso/farmacologia , Oligonucleotídeos Antissenso/metabolismo , Células Endoteliais/metabolismo , Células-Tronco Mesenquimais/metabolismo , Diferenciação Celular/genética , MicroRNAs/metabolismo
8.
Sensors (Basel) ; 23(7)2023 Mar 27.
Artigo em Inglês | MEDLINE | ID: mdl-37050545

RESUMO

The elastic light-scatter (ELS) technique, which detects and discriminates microbial organisms based on the light-scatter pattern of their colonies, has demonstrated excellent classification accuracy in pathogen screening tasks. The implementation of the multispectral approach has brought further advantages and motivated the design and validation of a hyperspectral elastic light-scatter phenotyping instrument (HESPI). The newly developed instrument consists of a supercontinuum (SC) laser and an acousto-optic tunable filter (AOTF). The use of these two components provided a broad spectrum of excitation light and a rapid selection of the wavelength of interest, which enables the collection of multiple spectral patterns for each colony instead of relying on single band analysis. The performance was validated by classifying microflora of green-leafed vegetables using the hyperspectral ELS patterns of the bacterial colonies. The accuracy ranged from 88.7% to 93.2% when the classification was performed with the scattering pattern created at a wavelength within the 473-709 nm region. When all of the hyperspectral ELS patterns were used, owing to the vastly increased size of the data, feature reduction and selection algorithms were utilized to enhance the robustness and ultimately lessen the complexity of the data collection. A new classification model with the feature reduction process improved the overall classification rate to 95.9%.


Assuntos
Bactérias , Elasticidade , Luz , Fenômenos Fisiológicos Bacterianos , Algoritmos
9.
Appl Spectrosc ; 77(6): 603-615, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-37097821

RESUMO

In this study, we propose a transfer learning-based classification model for identifying scrap metal using an augmented training dataset consisting of laser-induced breakdown spectroscopy (LIBS) measurement of standard reference material (SRMs) samples, considering varying experimental setups and environmental conditions. LIBS provides unique spectra for identifying unknown samples without complicated sample preparation. Thus, LIBS systems combined with machine learning methods have been actively studied for industrial applications such as scrap metal recycling. However, in machine learning models, a training set of the used samples may not cover the diversity of the scrap metal encountered in field measurements. Moreover, differences in experimental configuration, where laboratory standards and real samples are analyzed in situ, may lead to a wider gap in the distribution of training and test sets, dramatically reducing the performance of the LIBS-based fast classification system for real samples. To address these challenges, we propose a two-step Aug2Tran model. First, we augment the SRM dataset by synthesizing spectra of unobserved types through attenuation of dominant peaks corresponding to sample composition and generating spectra depending on the target sample using a generative adversarial network. Second, we used the augmented SRM dataset to build a robust real-time classification model with a convolutional neural network, which is further customized for the target scrap metal with limited measurements through transfer learning. For evaluation, SRMs of five representative metal types, including aluminum, copper, iron, stainless steel, and brass, are measured with a typical setup to form the SRM dataset. For testing, scrap metal from actual industrial fields is experimented with three different configurations, resulting in eight different test datasets. The experimental results show that the proposed scheme produces an average classification accuracy of 98.25% for the three experimental conditions, as high as the results of the conventional scheme with three separately trained and executed models. Additionally, the proposed model improves the classification accuracy of arbitrarily shaped static or moving samples with various surface contaminations and compositions, and even for differing ranges of charted intensities and wavelengths. Therefore, the proposed Aug2Tran model can be used as a systematic model for scrap metal classification with generalizability and ease of implementation.


Assuntos
Alumínio , Metais , Contaminação de Medicamentos , Análise Espectral , Lasers
10.
Sensors (Basel) ; 23(4)2023 Feb 14.
Artigo em Inglês | MEDLINE | ID: mdl-36850732

RESUMO

Real-time detection and disinfection of foodborne pathogens are important for preventing foodborne outbreaks and for maintaining a safe environment for consumers. There are numerous methods for the disinfection of hazardous organisms, including heat treatment, chemical reaction, filtration, and irradiation. This report evaluated a portable instrument to validate its simultaneous detection and disinfection capability in typical laboratory situations. In this challenging study, three gram-negative and two gram-positive microorganisms were used. For the detection of contamination, inoculations of various concentrations were dispensed on three different surface types to estimate the performance for minimum-detectable cell concentration. Inoculations higher than 103~104 CFU/mm2 and 0.15 mm of detectable contaminant size were estimated to generate a sufficient level of fluorescence signal. The evaluation of disinfection efficacy was conducted on three distinct types of surfaces, with the energy density of UVC light (275-nm) ranging from 4.5 to 22.5 mJ/cm2 and the exposure time varying from 1 to 5 s. The study determined the optimal energy dose for each of the microorganisms species. In addition, surface characteristics may also be an important factor that results in different inactivation efficacy. These results demonstrate that the proposed portable device could serve as an in-field detection and disinfection unit in various environments, and provide a more efficient and user-friendly way of performing disinfection on large surface areas.


Assuntos
Desinfecção , Filtração , Fenômenos Físicos , Surtos de Doenças , Contaminação de Medicamentos
11.
Sensors (Basel) ; 23(2)2023 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-36679567

RESUMO

A haptic sensor coupled to a gamepad or headset is frequently used to enhance the sense of immersion for game players. However, providing haptic feedback for appropriate sound effects involves specialized audio engineering techniques to identify target sounds that vary according to the game. We propose a deep learning-based method for sound event detection (SED) to determine the optimal timing of haptic feedback in extremely noisy environments. To accomplish this, we introduce the BattleSound dataset, which contains a large volume of game sound recordings of game effects and other distracting sounds, including voice chats from a PlayerUnknown's Battlegrounds (PUBG) game. Given the highly noisy and distracting nature of war-game environments, we set the annotation interval to 0.5 s, which is significantly shorter than the existing benchmarks for SED, to increase the likelihood that the annotated label contains sound from a single source. As a baseline, we adopt mobile-sized deep learning models to perform two tasks: weapon sound event detection (WSED) and voice chat activity detection (VCAD). The accuracy of the models trained on BattleSound was greater than 90% for both tasks; thus, BattleSound enables real-time game sound recognition in noisy environments via deep learning. In addition, we demonstrated that performance degraded significantly when the annotation interval was greater than 0.5 s, indicating that the BattleSound with short annotation intervals is advantageous for SED applications that demand real-time inferences.


Assuntos
Benchmarking , Som , Retroalimentação , Ruído , Audição
12.
Int J Mol Sci ; 24(1)2023 Jan 03.
Artigo em Inglês | MEDLINE | ID: mdl-36614321

RESUMO

Mesenchymal stromal cells derived from induced pluripotent stem cells (iMSCs) have been proposed as alternative sources of primary MSCs with various advantages for cell therapeutic trials. However, precise evaluation of the differences between iMSCs and primary MSCs is lacking due to individual variations in the donor cells, which obscure direct comparisons between the two. In this study, we generated donor-matched iMSCs from individual bone marrow-derived MSCs and directly compared their cell-autonomous and paracrine therapeutic effects. We found that the transition from primary MSCs to iMSCs is accompanied by a functional shift towards higher proliferative activity, with variations in differentiation potential in a donor cell-dependent manner. The transition from MSCs to iMSCs was associated with common changes in transcriptomic and proteomic profiles beyond the variations of their individual donors, revealing expression patterns unique for the iMSCs. These iMSC-specific patterns were characterized by a shift in cell fate towards a pericyte-like state and enhanced secretion of paracrine cytokine/growth factors. Accordingly, iMSCs exhibited higher support for the self-renewing expansion of primitive hematopoietic progenitors and more potent immune suppression of allogenic immune responses than MSCs. Our study suggests that iMSCs represent a separate entity of MSCs with unique therapeutic potential distinct from their parental MSCs, but points to the need for iMSC characterization in the individual basis.


Assuntos
Células-Tronco Pluripotentes Induzidas , Células-Tronco Mesenquimais , Proteômica , Diferenciação Celular/fisiologia , Transdução de Sinais , Células-Tronco Mesenquimais/metabolismo
13.
Foods ; 12(2)2023 Jan 14.
Artigo em Inglês | MEDLINE | ID: mdl-36673494

RESUMO

Laser-induced breakdown spectroscopy (LIBS) is an atomic-emission spectroscopy technique that employs a focused laser beam to produce microplasma. Although LIBS was designed for applications in the field of materials science, it has lately been proposed as a method for the compositional analysis of agricultural goods. We deployed commercial handheld LIBS equipment to illustrate the performance of this promising optical technology in the context of food authentication, as the growing incidence of food fraud necessitates the development of novel portable methods for detection. We focused on regional agricultural commodities such as European Alpine-style cheeses, coffee, spices, balsamic vinegar, and vanilla extracts. Liquid examples, including seven balsamic vinegar products and six representatives of vanilla extract, were measured on a nitrocellulose membrane. No sample preparation was required for solid foods, which consisted of seven brands of coffee beans, sixteen varieties of Alpine-style cheeses, and eight different spices. The pre-processed and standardized LIBS spectra were used to train and test the elastic net-regularized multinomial classifier. The performance of the portable and benchtop LIBS systems was compared and described. The results indicate that field-deployable, portable LIBS devices provide a robust, accurate, and simple-to-use platform for agricultural product verification that requires minimal sample preparation, if any.

14.
Vaccines (Basel) ; 12(1)2023 Dec 21.
Artigo em Inglês | MEDLINE | ID: mdl-38276669

RESUMO

To analyze the relationship between homologous and heterologous serological titers of immunized pigs and their protection statuses against FMD virus challenges, in the present study, the correlation between the virus neutralization titers at 21 and 28 dpv and the protection statuses at 28 dpv against challenge with FMD virus was analyzed using data sets comprising five different combinations of homologous or heterologous challenge experiments in pigs vaccinated with type O (n = 96), A (n = 69), and Asia 1 (n = 74). As a result, the experiments were divided into three groups (21D-1, 21D-2, and 21D-3) in the 21-dpv model and two groups (28D-1 and 28D-2) in the 28-dpv model. Each response curve of groups 21D-1 and 21D-2 in the 21-dpv model was very similar to each curve of groups 28D-1 and 28D-2 in the 28-dpv model, respectively, even though there was an exceptional extra group (21D-3) in the 21-dpv model. The average titers estimating 0.75 probability of protection ranged from 1.06 to 1.62 log10 in the 21-dpv model and from 1.26 to 1.64 log10 in the 28-dpv model. In summary, we demonstrated that the serological method is useful for predicting the homologous and heterologous protection statuses of vaccinated pigs.

15.
Int J Mol Sci ; 23(12)2022 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-35742968

RESUMO

Co-culture system, in which two or more distinct cell types are cultured together, is advantageous in that it can mimic the environment of the in vivo niche of the cells. In this study, we presented a strategy to analyze the secretome of a specific cell type under the co-culture condition in serum-supplemented media. For the cell-specific secretome analysis, we expressed the mouse mutant methionyl-tRNA synthetase for the incorporation of the non-canonical amino acid, azidonorleucine into the newly synthesized proteins in cells of which the secretome is targeted. The azidonorleucine-tagged secretome could be enriched, based on click chemistry, and distinguished from any other contaminating proteins, either from the cell culture media or the other cells co-cultured with the cells of interest. In order to have more reliable true-positive identifications of cell-specific secretory bodies, we established criteria to exclude any identified human peptide matched to bovine proteins. As a result, we identified a maximum of 719 secreted proteins in the secretome analysis under this co-culture condition. Last, we applied this platform to profile the secretome of mesenchymal stem cells and predicted its therapeutic potential on osteoarthritis based on secretome analysis.


Assuntos
Metionina tRNA Ligase , Animais , Bovinos , Química Click , Técnicas de Cocultura , Metionina tRNA Ligase/genética , Camundongos , Proteínas , Secretoma
16.
Anal Chem ; 94(10): 4192-4200, 2022 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-35239305

RESUMO

Phosphorylation is a crucial component of cellular signaling cascades. It controls a variety of biological cellular functions, including cell growth and apoptosis. Owing to the low stoichiometry of phosphorylated proteins, the enrichment of phosphopeptides prior to LC-MS/MS is necessary for comprehensive phosphoproteome analysis, and quantitative phosphoproteomic workflows are typically limited by the amount of sample required. To address this issue, we developed an easy-to-establish, widely applicable, and reproducible strategy to increase phosphoproteomic signals from a small amount of sample without a phosphoenrichment step. By exploiting the multiplexing nature of isobaric labeling to generate a merged signal from multiple samples, and using a larger amount of enriched phosphopeptides as a carrier, we were able to increase trace amounts of phosphopeptides in the unpurified sample to an identifiable level and perform quantification using the reporter ion intensity of the isobaric tag. Our results showed that >1400 phosphopeptides were quantified from 250 ng of tryptic peptides prepared from cells. In a proof-of-concept of our strategy, we distinguished three types of lung cancer cell lines based on their quantitative phosphoproteomic data and identified changes in the phosphoproteome induced by drug treatment.


Assuntos
Fosfopeptídeos , Proteômica , Cromatografia Líquida , Fosfopeptídeos/análise , Fosforilação , Proteoma/metabolismo , Proteômica/métodos , Espectrometria de Massas em Tandem/métodos
17.
Appl Microbiol Biotechnol ; 106(4): 1531-1542, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-35141866

RESUMO

The potential use of biological agents has become a major public health concern worldwide. According to the CDC classification, Bacillus anthracis and Clostridium botulinum, the bacterial pathogens that cause anthrax and botulism, respectively, are considered to be the most dangerous potential biological agents. Currently, there is no licensed vaccine that is well suited for mass immunization in the event of an anthrax or botulism epidemic. In the present study, we developed a dual-expression system-based multipathogen DNA vaccine that encodes the PA-D4 gene of B. anthracis and the HCt gene of C. botulinum. When the multipathogen DNA vaccine was administered to mice and guinea pigs, high level antibody responses were elicited against both PA-D4 and HCt. Analysis of the serum IgG subtype implied a combined Th1/Th2 response to both antigens, but one that was Th2 skewed. In addition, immunization with the multipathogen DNA vaccine induced effective neutralizing antibody activity against both PA-D4 and HCt. Finally, the protection efficiency of the multipathogen DNA vaccine was determined by sequential challenge with 10 LD50 of B. anthracis spores and 10 LD50 of botulinum toxin, or vice versa, and the multipathogen DNA vaccine provided higher than 50% protection against lethal challenge with both high-risk biothreat agents. Our studies suggest the strategy used for this anthrax-botulinum multipathogen DNA vaccine as a prospective approach for developing emergency vaccines that can be immediately distributed on a massive scale in response to a biothreat emergency or infectious disease outbreak. Key points • A novel multipathogen DNA vaccine was constructed against anthrax and botulism. • Robust immune responses were induced following vaccination. • Suggests a potential vaccine development strategy against biothreat agents.


Assuntos
Vacinas contra Antraz , Antraz , Bacillus anthracis , Botulismo , Vacinas de DNA , Animais , Antraz/prevenção & controle , Anticorpos Antibacterianos , Antígenos de Bactérias/genética , Bacillus anthracis/genética , Armas Biológicas , Botulismo/prevenção & controle , Cobaias , Imunidade , Camundongos , Vacinas de DNA/genética
18.
J Clin Med ; 10(8)2021 Apr 20.
Artigo em Inglês | MEDLINE | ID: mdl-33924095

RESUMO

The sources of mesenchymal stromal cells (MSCs) for cell therapy trials are expanding, increasing the need for their characterization. Here, we characterized multi-donor, turbinate-derived MSCs (TB-MSCs) that develop from the neural crest, and compared them to bone marrow-derived MSCs (BM-MSCs). TB-MSCs had higher proliferation potential and higher self-renewal of colony forming cells, but lower potential for multi-lineage differentiation than BM-MSCs. TB-MSCs expressed higher levels of neural crest markers and lower levels of pericyte-specific markers. These neural crest-like properties of TB-MSCs were reflected by their propensity to differentiate into neuronal cells and proliferative response to nerve growth factors. Proteomics (LC-MS/MS) analysis revealed a distinct secretome profile of TB-MSCs compared to BM and adipose tissue-derived MSCs, exhibiting enrichments of factors for cell-extracellular matrix interaction and neurogenic signaling. However, TB-MSCs and BM-MSCs exhibited comparable suppressive effects on the allo-immune response and comparable stimulatory effects on hematopoietic stem cell self-renewal. In contrast, TB-MSCs stimulated growth and metastasis of breast cancer cells more than BM-MSCs. Altogether, our multi-donor characterization of TB-MSCs reveals distinct cell autonomous and paracrine properties, reflecting their unique developmental origin. These findings support using TB-MSCs as an alternative source of MSCs with distinct biological characteristics for optimal applications in cell therapy.

19.
Food Chem ; 352: 129329, 2021 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-33684719

RESUMO

A simple, novel, rapid, and non-destructive spectroscopic method that employs the deep spectral network for beef-freshness classification was developed. The deep-learning-based model classified beef freshness by learning myoglobin information and reflectance spectra over different freshness states. The reflectance spectra (480-920 nm) were measured from 78 beef samples for 17 days, and the datasets were sorted into three freshness classes based on their pH values. Myoglobin information showed statistically significant differences depending on the freshness; consequently, it was utilized as a crucial parameter for classification. The model exhibited improved performance when the reflectance spectra were combined with the myoglobin information. The accuracy of the proposed model improved to 91.9%, whereas that of the single-spectra model was 83.6%. Further, a high value for the area under the receiver operating characteristic curve (0.958) was recorded. This study provides a basis for future studies on the investigation of myoglobin information associated with meat freshness.


Assuntos
Aprendizado Profundo , Qualidade dos Alimentos , Mioglobina/química , Carne Vermelha/classificação , Análise Espectral , Animais , Bovinos , Mioglobina/análise , Carne Vermelha/análise
20.
Int J Mol Sci ; 22(2)2021 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-33467726

RESUMO

Mesenchymal stem cells (MSCs) have the potential to be a viable therapy against various diseases due to their paracrine effects, such as secretion of immunomodulatory, trophic and protective factors. These cells are known to be distributed within various organs and tissues. Although they possess the same characteristics, MSCs from different sources are believed to have different secretion potentials and patterns, which may influence their therapeutic effects in disease environments. We characterized the protein secretome of adipose (AD), bone marrow (BM), placenta (PL), and Wharton's jelly (WJ)-derived human MSCs by using conditioned media and analyzing the secretome by mass spectrometry and follow-up bioinformatics. Each MSC secretome profile had distinct characteristics depending on the source. However, the functional analyses of the secretome from different sources showed that they share similar characteristics, such as cell migration and negative regulation of programmed cell death, even though differences in the composition of the secretome exist. This study shows that the secretome of fetal-derived MSCs, such as PL and WJ, had a more diverse composition than that of AD and BM-derived MSCs, and it was assumed that their therapeutic potential was greater because of these properties.


Assuntos
Tecido Adiposo/citologia , Células-Tronco Mesenquimais/metabolismo , Placenta/citologia , Cordão Umbilical/citologia , Geleia de Wharton/citologia , Medula Óssea , Células da Medula Óssea/citologia , Diferenciação Celular , Proliferação de Células , Cromatografia Líquida , Análise por Conglomerados , Técnicas de Cocultura , Biologia Computacional , Meios de Cultivo Condicionados , Meios de Cultura Livres de Soro , Feminino , Humanos , Espectrometria de Massas , Osteogênese , Gravidez , Proteômica , Espectrometria de Massas em Tandem
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