Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 1.857
Filtrar
1.
Food Res Int ; 164: 112361, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36737949

RESUMO

This study investigated the transport stress (crowding stress and duration) on the physicochemical properties, energy metabolism and antioxidant enzyme activities of the red swamp crayfish (Procambarus clarkii) tail muscle (CTM). Besides, transcriptomic and metabolomic were conducted to elucidate the possible mechanism of CTM alternations during transport stress. The survival rate of crayfish gradually decreased with the external crowding stress and crowding time increasing. The transport stress also led to the increased distance among muscle fibers, water mobility and energy consumption, and the decreased of water holding capacity (WHC), hardness of CTM. The hepatopancreas exhibited more sensitive to crowding stress than muscle. The multi-omics analysis revealed that transport stress could interfere the translation and protein folding functions of ribosomal proteins, fatty acid metabolism and degradation, physiological functions of mitochondria in CTM. This study could provide critical information to increase the understanding of the regulation mechanism of crayfish when subjected to transport stress.

2.
Brain Behav ; : e2890, 2023 Feb 03.
Artigo em Inglês | MEDLINE | ID: mdl-36738135

RESUMO

BACKGROUND: Ischemic stroke is a clinical emergency caused by insufficient intracranial blood supply, which eventually leads to brain tissue necrosis and neurological impairment. Predictive nursing intervention has achieved impressive success in the nursing of multiple surgeries. However, the role of predictive nursing intervention in the care of patients with ischemic stroke remains unclear. METHODS: This study was a randomized controlled trial. Based on the inclusion and exclusion criteria, 126 patients were randomly assigned into two groups, namely the control group and the predictive nursing intervention group. Both groups were treated with thrombolytic therapy with alteplase. The patients in the control group were given routine nursing intervention and the predictive nursing intervention group received additional predictive care. Neurologic functions and cognitive impairment were evaluated by National Institutes of Health Stroke Scale (NIHSS), Fugl-Meyer assessment (FMA), Montreal cognitive assessment (MoCA), and mini-mental state examination (MMSE) scales, respectively. Door-to-Needle Times, venous thromboembolism (VTE)-related parameters, and complications were recorded. RESULTS: Predictive nursing intervention significantly shortened the Door-to-Needle Times and enhanced the peak/average femoral venous blood flow and femoral venous diameter. In addition, predictive nursing intervention improved the NIHSS, FMA, MMSE, and MoCA scores and remarkably reduced the recurrence of ischemic stroke, deep vein thrombosis and gingival bleeding. CONCLUSION: Predictive nursing intervention is beneficial to improve the effects of thrombolytic therapy in patients with ischemic stroke, which improves the neurological, cognitive and motor functions of patients, and reduces the occurrence of complications, suggesting an important clinical application value.

3.
Eur Radiol ; 2023 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-36725721

RESUMO

OBJECTIVES: To evaluate the clinical value of contrast-enhanced ultrasound (CEUS) with time-intensity curve (TIC) in distinguishing different types of hypovascular solid pancreatic lesions. METHODS: A total of 89 patients with 90 pancreatic lesions (all confirmed by surgery or biopsy pathology) that manifested hypoenhancement on contrast-enhanced ultrasound (CEUS) were included in this study. Six peak enhancement patterns were proposed for differentiating hypovascular pancreatic lesions. CEUS qualitative and TIC-based quantitative parameters were analyzed, and each lesion was scored based on the statistically significant qualitative parameters to evaluate the diagnostic ability of CEUS for hypovascular solid pancreatic lesions. RESULTS: Qualitative parameters such as peak enhancement pattern II/III/IV, penetrating vessels, centripetal enhancement, and early washout were reliable indicators of malignant lesions, and lesions scored based on these qualitative parameters, with a score ≥ 2, were highly suspected to be malignant lesions. Pattern I had an accuracy of 83.33% for predicting mass-forming pancreatitis (MFP), pattern V had an accuracy of 96.67% for predicting solid pseudopapillary tumors of the pancreas (SPTP), and pattern VI had an accuracy of 81.11% for predicting neuroendocrine tumors/carcinomas (NETs/NECs). For quantitative analysis, nodule/pancreatic parenchyma echo intensity reduction ratio was significantly greater in malignant lesions. CONCLUSIONS: CEUS qualitative and TIC-based quantitative parameters have clinical value in distinguishing malignant from benign hypovascular pancreatic lesions. KEY POINTS: • Contrast-enhanced ultrasound helps clinicians assess patients with pancreatic lesions. • Six peak enhancement patterns are proposed for differentiating pancreatic hypovascular lesions. • Qualitative parameters such as peak enhancement pattern II/III/IV, penetrating vessels, centripetal enhancement, early washout, and quantitative parameter nodule/pancreatic parenchyma echo intensity reduction ratio were important characteristics to discriminate malignant from hypovascular benign lesions.

4.
Food Chem ; 411: 135386, 2023 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-36652882

RESUMO

This paper investigated the effects of Lentinus edodes stipes insoluble dietary fiber (LESIDF, 0%-3.0%) on the quality and microscopic properties of pork myofibrillar protein (MP) gels. The results showed that the water holding capacity and gel strength of composite gels enhanced with increasing LESIDF (1.0%-2.5%), and reached the maximum at the level of 2.5%-3.0%. Disulfide and non-disulfide covalent bonds were major chemical forces maintaining the 3D network of LESIDF-MP composite gels. LESIDF also promoted the formation of ionic and hydrogen bonds, confirmed by the self-assembly of ß-sheets to α-helices, leading to a compact gel network structure. The observation of paraffin section revealed that LESIDF could capture more water molecules in gels, which was consistent with the transformation of free water to immobilized water. Overall, the optimal addition of LESIDF was 2.5%-3.0%, which provided a good strategy for LESIDF as an agricultural by-product to improve the quality of gel meat products.

5.
Acad Radiol ; 2023 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-36610930

RESUMO

OBJECTIVES: To prolong the survival, the value of a computed tomography-based radiomic score (RS) in stratifying survival and guiding personalized chemotherapy strategies in far-advanced gastric cancer (FGC) was investigated. MATERIALS AND METHODS: This retrospective multicenter study enrolled 283 FGC patients (cT4a/bNxM0-1) from three centers. Patients from one center were randomly divided into the training (n = 166) and internal validation (n = 83) cohorts, whereas the external validation cohort (n = 34) consisted of patients from the two other centers. The RS was calculated for each patient to predict progression-free survival (PFS). Features from the primary tumor and main metastasis (peritoneum, liver, and lymph node) were integrated in the training cohort and then validated for its ability to stratify PFS and overall survival (OS) in the validation cohort. The association between the RS and efficacy of neoadjuvant intraperitoneal and systemic (NIPS) therapy was also explored. RESULTS: The RS demonstrated a favorable prognostic ability to predict PFS in all cohorts (training: C-index 0.83, 95% confidence interval [CI]: 0.788-0.872; internal validation: C-index 0.75, 95% CI: 0.682-0.818; external validation: C-index 0.76, 95% CI: 0.669-0.851; all p < 0.05), as well as an excellent ability to stratify the PFS and OS in both the whole population and metastatic subgroups (p < 0.05). Patients with a low score were more likely to undergo surgery after perioperative chemotherapy (p < 0.05). Furthermore, only high-scoring patients with peritoneal metastasis benefited from NIPS. CONCLUSION: The RS may be an effective risk stratifier for the outcomes of FGC patients and may be used to select patients who can benefit from NIPS therapy.

6.
Fish Shellfish Immunol ; 134: 108559, 2023 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-36681364

RESUMO

Antibacterial peptide (AMP), an effector of the innate immune system, is an essential component of invertebrate innate immunity. Crustin is a family of antimicrobial peptides that are widely studied in crustaceans. Here we report a novel crustin (designated Shcrustin) from the freshwater crab Sinopotamon henanense. The results revealed that the full-length cDNA of Shcrustin was 691 bp with an open reading frame (ORF) of 510 bp. Phylogenetic analysis of the Shcrustin sequence showed that it clustered with type II crustin. Shcrustin exists in different tissues, among which the highest expression level is found in the gills. After the bacterial challenge, the expression of Shcrustin increased in hemocytes or gills. However, crustin expression was suppressed in the presence of cadmium (Cd). To elucidate the biological activity of Shcrustin, we constructed a recombinant Shcrustin protein. Purified rShcrustin could bind to a variety of bacteria and inhibit the growth of different bacteria indicating that Shcrustin has inhibitory activity against gram-positive and gram-negative bacteria. In addition, the phagocytic rate of hemocytes toward bacteria decreased after the interference of Shcrustin expression by RNA interference, suggesting that Shcrustin may be involved in such a process. Therefore, we conclude that Shcrustin may be involved in the innate immunity of S. henanense by binding to bacteria and promoting hemolymph phagocytosis to clear invading pathogens. It is an important immune effector against pathogen infection. In the presence of Cd, it may alter the expression of Shcrustin and suppress its immune function.

7.
Fish Shellfish Immunol ; : 108565, 2023 Jan 23.
Artigo em Inglês | MEDLINE | ID: mdl-36702328

RESUMO

Prophenoloxidase (proPO) is essential in the prophenoloxidase-activating system (proPO-AS) which is important for defense against foreign infection in crustaceans. However, most studies have focused on expression in the presence of a single pathogenic bacterium, and very few have addressed the presence of environmental contaminants simultaneously, such as cadmium (Cd) and Aeromonas hydrophila. Our study aimed to investigate the function of proPO in the freshwater crab Sinopotamon henanense and the changes in its expression by Cd and infection of A. hydrophila. A novel proPO from the hemocytes of S. henanense (ShproPO) was found in this research, the full-length cDNA of ShproPO was 2620 bp of encoding a protein of 678 amino acids containing three typical hemocyanin domains. The ShproPO protein could be found in both the granular (GHc) and the semi-granular hemocytes (SGHc). The ShproPO mRNA was found to be abundantly expressed in hemocytes and could be influenced by A. hydrophila infection. These results indicate that ShproPO could be involved in the antibacterial process. Further research found that low concentrations of Cd could promote its expression after infection with A. hydrophila. Therefore, it was hypothesized that Cd disrupted the response of crabs to A. hydrophila infection. Subsequently, PO enzyme activity was found to be significantly reduced through in vivo RNA interference with ShproPO, and the results suggested that ShproPO is likely to be a key enzyme in the melanization response. Finally, ShproPO was found to significantly enhance the phagocytosis of A. hydrophila-infected hemocytes by in vitro recombination, confirming that ShproPO is involved in hemocyte-mediated melanization and phagocytosis. Our findings reveal completely new insight into the immunotoxicity of Cd and the immune function of ShproPO in S. henanense.

8.
Food Chem ; 410: 135353, 2023 Jun 01.
Artigo em Inglês | MEDLINE | ID: mdl-36608548

RESUMO

This study investigated the influence of pile fermentation on the physicochemical, functional, and biological properties of tea polysaccharides (TPS). Results indicated that the extraction yield, uronic acid content, and polyphenol content of TPS greatly increased from 1.8, 13.1 and 6.3 % to 4.1, 27.9, and 7.8 %, respectively, but the molecular weight markedly decreased from 153.7 to 76.0 kDa after pile fermentation. Additionally, the interfacial, emulsion formation, and emulsion stabilization properties of TPS were significantly improved after pile fermentation. For instance, 1.0 wt% TPS isolated from dark tea (D-TPS) can fabricate 8.0 wt% MCT oil-in-water nanoemulsion (d32 ≈ 159 nm) with potent storage stability. Moreover, the antioxidant and α-glucosidase inhibitory activities of D-TPS was higher than that of TPS isolated from sun-dried raw tea (R-TPS). Overall, this study indicated that pile fermentation markedly affected the physicochemical and structural characteristics of TPS, thereby improving their functional and biological properties.

9.
Cell Discov ; 9(1): 3, 2023 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-36609558

RESUMO

SARS-CoV-2 Omicron subvariants have demonstrated extensive evasion from monoclonal antibodies (mAbs) developed for clinical use, which raises an urgent need to develop new broad-spectrum mAbs. Here, we report the isolation and analysis of two anti-RBD neutralizing antibodies BA7208 and BA7125 from mice engineered to produce human antibodies. While BA7125 showed broadly neutralizing activity against all variants except the Omicron sublineages, BA7208 was potently neutralizing against all tested SARS-CoV-2 variants (including Omicron BA.1-BA.5) except Mu. By combining BA7208 and BA7125 through the knobs-into-holes technology, we generated a biparatopic antibody BA7208/7125 that was able to neutralize all tested circulating SARS-CoV-2 variants. Cryo-electron microscopy structure of these broad-spectrum antibodies in complex with trimeric Delta and Omicron spike indicated that the contact residues are highly conserved and had minimal interactions with mutational residues in RBD of current variants. In addition, we showed that administration of BA7208/7125 via the intraperitoneal, intranasal, or aerosol inhalation route showed potent therapeutic efficacy against Omicron BA.1 and BA.2 in hACE2-transgenic and wild-type mice and, separately, effective prophylaxis. BA7208/7125 thus has the potential to be an effective candidate as an intervention against COVID-19.

10.
Sci Rep ; 13(1): 1225, 2023 Jan 21.
Artigo em Inglês | MEDLINE | ID: mdl-36681777

RESUMO

Idiopathic pulmonary fibrosis (IPF) is a progressive interstitial lung disease that causes irreversible damage to lung tissue characterized by excessive deposition of extracellular matrix (ECM) and remodeling of lung parenchyma. The current diagnosis of IPF is complex and usually completed by a multidisciplinary team including clinicians, radiologists and pathologists they work together and make decision for an effective treatment, it is imperative to introduce novel practical methods for IPF diagnosis. This study provided a new diagnostic model of idiopathic pulmonary fibrosis based on machine learning. Six genes including CDH3, DIO2, ADAMTS14, HS6ST2, IL13RA2, and IGFL2 were identified based on the differentially expressed genes in IPF patients compare to healthy subjects through a random forest classifier with the existing gene expression databases. An artificial neural network model was constructed for IPF diagnosis based these genes, and this model was validated by the distinctive public datasets with a satisfactory diagnostic accuracy. These six genes identified were significant correlated with lung function, and among them, CDH3 and DIO2 were further determined to be significantly associated with the survival. Putting together, artificial neural network model identified the significant genes to distinguish idiopathic pulmonary fibrosis from healthy people and it is potential for molecular diagnosis of IPF.


Assuntos
Fibrose Pulmonar Idiopática , Doenças Pulmonares Intersticiais , Humanos , Fibrose Pulmonar Idiopática/diagnóstico , Fibrose Pulmonar Idiopática/genética , Pulmão , Sulfotransferases
11.
Health Place ; 79: 102971, 2023 Jan 20.
Artigo em Inglês | MEDLINE | ID: mdl-36682263

RESUMO

Automobile dependence and physical inactivity have become common health challenges for residents in large suburban residential areas. Limited literature has examined the associations between the built environment and active travel in such residential areas and the differences in these associations among residents from different neighborhoods. To avoid inaccurate results potentially derived from residence-based measures, we adopt a mobility-based approach for environmental exposure assessment. Using GPS data from 530 trips made by 98 participants in a large residential area in Shanghai, we investigate the relationships between neighborhood types, pollution perceptions, built environment features and active travel. The results indicate that residents in affordable and relocation housing make fewer active trips than those in market-rate housing, while the built environment seems to mitigate this difference. Sports facilities promote active travel while commercial facilities and road intersections discourage it. We identify significant interactions between the percentage of green space and neighborhood type, as well as floor area ratio and air pollution perception. Interventions promoting active travel include active-travel-friendly design for commercial facilities and road intersections, the provision of more sports facilities, a careful increase in floor area ratio, and the provision of more green space that is attractive to residents from different neighborhoods.

12.
J Exp Child Psychol ; 228: 105619, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36592578

RESUMO

Many cross-sectional studies have shown that number line estimation is associated with other mathematical skills, but there has been limited longitudinal research. To systematically examine such associations longitudinally at the earliest stage of mathematical learning, the current study tested 40 Chinese preschoolers (mean age = 4.97 years, SD = 0.18) and followed them up 8 months later. For both waves of data collection, children were administered six tasks: number line estimation, dot counting, comparison of two dot arrays, comparison of triple dot arrays, symbolic number comparison, and simple addition. Results of two-wave cross-lagged panel analysis showed that (a) dot counting and non-symbolic numerical comparison at Time 1 had significant longitudinal associations with number line estimation at Time 2, (b) number line estimation had bidirectional associations with symbolic number comparison, and (c) number line estimation at Time 1 had a marginally significant longitudinal association with simple addition at Time 2. These results extend the small but accumulating literature on the longitudinal relations between number line estimation and other mathematical skills and specify the important role of number line estimation in the early development of mathematical skills.


Assuntos
Cognição , Humanos , Pré-Escolar , Estudos Transversais , Aprendizagem , Matemática
13.
Chemosphere ; 316: 137863, 2023 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-36649895

RESUMO

Glufosinate-ammonium, the second largest transgene crop resistant herbicide, is classified as a mobile persistent pollutant by the U.S. Environmental Protection Agencybecause of its slow decomposition and easy mobile transfer in a water environment. The chronic and multigeneration toxicity of this compound to environmental organisms are alarming. In this study, racemic glufosinate-ammonium and the effective isomer, l-glufosinate-ammonium, were used as the test agents. The developmental, neurotoxic and reproductive toxicities of Caenorhabditis elegans to their parents and progeny were studied by continuous exposure in water at concentrations of 0.1, 1, 10 and 100 µg/L. The causes of toxicity differences were analysed from oxidative stress and transcription levels. Through oxidative stress of C. elegans, racemic glufosinate-ammonium and l-glufosinate-ammonium both mediated the developmental toxicity (shortened developmental cycle, reduced body length and width, promoted ageingand decreased longevity), neurotoxicity (inhibited head swinging, body bending frequency and acetylcholinesterase [AchE] activity) and reproductive toxicity (significant reductions in the number of eggs and offspring in vivo and induced apoptosis of gonadal cells). These phenomena caused oxidative damage (protein and membrane lipid peroxidation) and further induced apoptosis. The changes in various indicators caused by racemic glufosinate-ammonium exposure were more significant than those caused by l-glufosinate-ammonium exposure, and the reproduction-related indicators were more significant than the developmental and neurological indicators. A continuous accumulation of toxicity was observed after multiple generations of continuous exposure. These research results provide a data reference for the ecotoxicological evaluation and risk assessment of glufosinate-ammonium and contribute to the revision and improvement of the related environmental policies of glufosinate-ammonium.


Assuntos
Acetilcolinesterase , Caenorhabditis elegans , Animais , Caenorhabditis elegans/genética , Aminobutiratos/toxicidade , Reprodução
14.
Anal Chem ; 95(4): 2548-2560, 2023 Jan 31.
Artigo em Inglês | MEDLINE | ID: mdl-36656605

RESUMO

Imaged capillary isoelectric focusing (icIEF) and ion-exchange chromatography (IEX) are two essential techniques that are routinely used for charge variant analysis of therapeutic monoclonal antibodies (mAbs) during their development and in quality control. These two techniques that separate mAb charge variants based on different mechanisms and IEX have been developed as front-end separation techniques for online mass spectrometry (MS) detection, which is robust for intact protein identification. Recently, an innovative, coupled icIEF-MS technology has been constructed for protein charge variant analysis in our laboratory. In this study, icIEF-MS developed and strong cation exchange (SCX)-MS were optimized for charge heterogeneity characterization of a diverse of mAbs and their results were compared based on methodological validation. It was found that icIEF-MS outperformed SCX-MS in this study by demonstrating outstanding sensitivity, low carryover effect, accurate protein identification, and higher separation resolution although SCX-MS contributed to higher analysis throughput. Ultimately, integrating our novel icIEF-HRMS analysis with the more common SCX-MS can provide a promising and comprehensive strategy for accelerating the development of complex protein therapeutics.


Assuntos
Anticorpos Monoclonais , Anticorpos Monoclonais/química , Espectrometria de Massas/métodos , Focalização Isoelétrica/métodos , Cromatografia por Troca Iônica/métodos
15.
Chin J Nat Med ; 21(1): 58-64, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36641233

RESUMO

Drimane-type sesquiterpenoids are widely distributed in fungi. From the ethyl acetate extract of the earwig-derived Aspergillus sp. NF2396, seven new drimane-type sesquiterpenoids, named drimanenoids A-G (1-7), were isolated. Their structures were elucidated by diverse spectroscopic analysis including high-resolution ESI-MS, one- and two-dimensional NMR spectroscopy. Drimanenoids A-F (1-6) are new members of drimane-type sesquiterpenoid esterified with unsaturated fatty acid side chain at C-6. Drimanenoids C (3), D (4) and F (6) showed antibacterial activity against five types of bacteria with different inhibition diameters. Drimanenoid D (4) exhibited moderate cytotoxicity against human myelogenous leukemia cell line K562 with an IC50 value of 12.88 ± 0.11 µmol·L-1.


Assuntos
Sesquiterpenos , Humanos , Sesquiterpenos Policíclicos , Sesquiterpenos/química , Aspergillus/química , Espectroscopia de Ressonância Magnética , Estrutura Molecular
16.
Plant Cell ; 2023 Jan 10.
Artigo em Inglês | MEDLINE | ID: mdl-36625683

RESUMO

Elicitins are a large family of secreted proteins in Phytophthora. Clade 1 elicitins were identified decades ago as potent elicitors of immune responses in Nicotiana species, but the mechanisms underlying elicitin recognition are largely unknown. Here we identified an elicitin receptor in Nicotiana benthamiana that we named REL for Responsive to ELicitins. REL is a receptor-like protein (RLP) with an extracellular leucine-rich repeat (LRR) domain that mediates Phytophthora resistance by binding elicitins. Silencing or knocking out REL in N. benthamiana abolished elicitin-triggered cell death and immune responses. Domain deletion and site-directed mutagenesis revealed that the island domain (ID) located within the LRR domain of REL is crucial for elicitin recognition. In addition, sequence polymorphism in the ID underpins the genetic diversity of REL homologs in various Nicotiana species in elicitin recognition and binding. Remarkably, REL is phylogenetically distant from the elicitin response (ELR) protein, a LRR-RLP that was previously identified in the wild potato species Solanum microdontum and REL and ELR differ in the way they bind and recognize elicitins. Our findings provide insights into the molecular basis of plant innate immunity and highlight a convergent evolution of immune receptors towards perceiving the same elicitor.

17.
Ann Med ; 55(1): 342-347, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36598144

RESUMO

Aim: Tuberculous meningitis is an infectious disease of the central nervous system caused by Mycobacterium tuberculosis (M. tuberculosis). It mainly involves the meninges and brain parenchyma, as well as the spinal cord and meninges; Disability and mortality rates are high. In recent years, due to the increase of drug-resistant tuberculosis patients, population mobility and the prevalence of acquired immune deficiency syndrome, the incidence rate of tuberculosis has increased significantly, and tuberculous meningitis has also increased.Methods: At present, tuberculosis is still a worldwide infectious disease that seriously threatens human health, especially in underdeveloped and developing countries. China is the largest developing country in the world with a large population.Results: The situation of tuberculosis prevention and control is grim. Its disability rate is the highest in tuberculosis infection. In addition to the common non-specific manifestations, tuberculous meningoencephalitis may also have rare manifestations of stroke, hearing loss and visual loss.Conclusion: Understanding and timely improvement of corresponding examinations and targeted treatment will help improve the prognosis of patients.


Assuntos
Meningoencefalite , Mycobacterium tuberculosis , Tuberculose Meníngea , Humanos , Tuberculose Meníngea/complicações , Tuberculose Meníngea/diagnóstico , Tuberculose Meníngea/epidemiologia , Encéfalo , Meningoencefalite/epidemiologia , China
18.
Anal Biochem ; 663: 115034, 2023 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-36586502

RESUMO

Because the spike (S) protein of the severe acute respiratory syndrome coronavirus (SARS-CoV) is the immunodominant antigen, the S protein and its receptor-binding domain (RBD) are both targets currently to be genetically engineered for designing the broad-spectrum vaccine. In theory, the expressed protein exists as a set of variants that are roughly the same but slightly different, which depends on the protein expression system. The variants can be phenotypically manifested as charge heterogeneity. Here, we attempted to depict the charge heterogeneity of the trimeric SARS-CoV-2 RBD by using capillary isoelectric focusing with whole-column imaging detection (cIEF-WCID). In its nature form, the electropherogram fingerprints of the trimeric RBD were presented under optimized experimental conditions. The peaks of matrix buffers can be fully distinguishable from peaks of trimeric RBD. The isoelectric point (pI) was determined to be within a range of 6.67-9.54 covering the theoretical pI of 9.02. The fingerprints of three batches of trimeric RBDs are completely the same, with the intra-batch and batch-to-batch relative standard deviations (RSDs) of both pI values and area percentage of each peak no more than 1.0%, indicating that the production process is stable and this method can be used to surveillance the batch-to-batch consistency. The fingerprint remained unchanged after incubating at 37 °C for 7 d and oxidizing by 0.015% H2O2. In addition, the fingerprint was destroyed when adjusting the pH value to higher than 10.0 but still stable when the pH was lower than 4.0. In summary, the cIEF-WCID fingerprint can be used for the identification, batch-to-batch consistency evaluation, and stability study of the trimeric SARS-CoV-2 RBD, as part of a quality control strategy during the potential vaccine production.


Assuntos
COVID-19 , SARS-CoV-2 , Humanos , Focalização Isoelétrica/métodos , Peróxido de Hidrogênio , Ligação Proteica
19.
Exp Eye Res ; 226: 109352, 2023 01.
Artigo em Inglês | MEDLINE | ID: mdl-36528083

RESUMO

Müller cells, the glial cells of the retina, provide metabolic support for photoreceptors and inner retinal neurons, and have been proposed as source of the significant lactate production of this tissue. To better understand the role of lactate in retinal metabolism, we expressed a lactate and a glucose nanosensor in organotypic mouse retinal explants cultured for 14 days, and used FRET imaging in acute vibratome sections of the explants to study metabolite flux in real time. Pharmacological manipulation with specific monocarboxylate transporter (MCT) inhibitors and immunohistochemistry revealed the functional expression of MCT1, MCT2 and MCT4 in Müller cells of retinal explants. The introduction of FRET nanosensors to measure key metabolites at the cellular level may contribute to a better understanding of heretofore poorly understood issues in retinal metabolism.


Assuntos
Células Ependimogliais , Transferência Ressonante de Energia de Fluorescência , Camundongos , Animais , Células Ependimogliais/metabolismo , Proteínas de Membrana Transportadoras/metabolismo , Retina/metabolismo , Ácido Láctico/metabolismo , Transportadores de Ácidos Monocarboxílicos/metabolismo
20.
Front Public Health ; 10: 1048108, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36457331

RESUMO

Objective: Human adenovirus (HAdV) coinfection with other respiratory viruses is common, but adenovirus infection combined with human coronavirus-229E (HCoV-229E) is very rare. Study design and setting: Clinical manifestations, laboratory examinations, and disease severity were compared between three groups: one coinfected with HAdV-Ad7 and HCoV-229E, one infected only with adenovirus (mono-adenovirus), and one infected only with HCoV-229E (mono-HCoV-229E). Results: From July to August 2019, there were 24 hospitalized children: two were coinfected with HAdV-Ad7 and HCoV-229E, and 21 were infected with a single adenovirus infection. Finally, one 14-year-old boy presented with a high fever, but tested negative for HAdV-Ad7 and HCoV-229E. Additionally, three adult asymptotic cases with HCoV-229E were screened. No significant difference in age was found in the coinfection and mono-adenovirus groups (11 vs. 8 years, p = 0.332). Both groups had the same incubation period (2.5 vs. 3 days, p = 0.8302), fever duration (2.5 vs. 2.9 days, p = 0.5062), and length of hospital stay (7 vs. 6.76 days, p = 0.640). No obvious differences were found in viral loads between the coinfection and mono-adenovirus groups (25.4 vs. 23.7, p = 0.570), or in the coinfection and mono-HCoV-229E groups (32.9 vs. 30.06, p = 0.067). All cases recovered and were discharged from the hospital. Conclusion: HAdV-Ad7 and HCoV-229E coinfection in healthy children may not increase the clinical severity or prolong the clinical course. The specific interaction mechanism between the viruses requires further study.


Assuntos
Adenovírus Humanos , Coinfecção , Coronavirus , Adulto , Masculino , Criança , Humanos , Idoso de 80 Anos ou mais , Carga Viral , Hospitais
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA
...