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1.
Food Chem ; 451: 139410, 2024 Apr 20.
Artigo em Inglês | MEDLINE | ID: mdl-38670024

RESUMO

Dipicolinic acid (DPA), as a biomarker for Bacillus anthracis, is highly toxic at trace levels. Rapid and on-site quantitative detection of DPA is essential for maintaining food safety and public health. This work develops a dual-channel self-calibrated fluorescence sensor constructed by the YVO4:Eu and Tb-ß-diketone complex for rapid visual detection of DPA. This sensor exhibits high selectivity, fast response time, excellent detection sensitivity, and the detection limit is as low as 4.5 nM in the linear range of 0-16 µM. A smartphone APP and portable ultraviolet lamp can assemble a mobile fluorescence sensor for on-site analysis. Interestingly, adding Cu2+ ions can quench the fluorescence intensity of Tb3+. In contrast, the addition of cysteine can restore the fluorescence, allowing the accurate detection of Cu2+ ions and cysteine in environmental water and food samples. This work provides a portable sensor that facilitates real-time analysis of multiple targets in food and the environment.

2.
Neural Regen Res ; 19(9): 2027-2035, 2024 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-38227532

RESUMO

JOURNAL/nrgr/04.03/01300535-202409000-00035/figure1/v/2024-01-16T170235Z/r/image-tiff Neonatal hypoxic-ischemic encephalopathy is often associated with permanent cerebral palsy, neurosensory impairments, and cognitive deficits, and there is no effective treatment for complications related to hypoxic-ischemic encephalopathy. The therapeutic potential of human placental chorionic plate-derived mesenchymal stem cells for various diseases has been explored. However, the potential use of human placental chorionic plate-derived mesenchymal stem cells for the treatment of neonatal hypoxic-ischemic encephalopathy has not yet been investigated. In this study, we injected human placental chorionic plate-derived mesenchymal stem cells into the lateral ventricle of a neonatal hypoxic-ischemic encephalopathy rat model and observed significant improvements in both cognitive and motor function. Protein chip analysis showed that interleukin-3 expression was significantly elevated in neonatal hypoxic-ischemic encephalopathy model rats. Following transplantation of human placental chorionic plate-derived mesenchymal stem cells, interleukin-3 expression was downregulated. To further investigate the role of interleukin-3 in neonatal hypoxic-ischemic encephalopathy, we established an in vitro SH-SY5Y cell model of hypoxic-ischemic injury through oxygen-glucose deprivation and silenced interleukin-3 expression using small interfering RNA. We found that the activity and proliferation of SH-SY5Y cells subjected to oxygen-glucose deprivation were further suppressed by interleukin-3 knockdown. Furthermore, interleukin-3 knockout exacerbated neuronal damage and cognitive and motor function impairment in rat models of hypoxic-ischemic encephalopathy. The findings suggest that transplantation of hpcMSCs ameliorated behavioral impairments in a rat model of hypoxic-ischemic encephalopathy, and this effect was mediated by interleukin-3-dependent neurological function.

3.
Biochem Genet ; 2024 Jan 25.
Artigo em Inglês | MEDLINE | ID: mdl-38273154

RESUMO

To investigate the effect and potential mechanism of human-derived urine stem cells (hUSCs) in inhibiting retinal aging by using experimental and bioinformatics. Retinal pigment epithelial cells cultured in vitro, which were randomly divided into normal group, aging group and supernatant of hUSCs group. Cell counting kit-8 detection, senescence-related ß-galactosidase, and Annexin V/PI staining were performed to detect cell viability, senescence, and apoptosis. Subsequently, bioinformatics methods were used to explore the underlying mechanisms, in which, targets both hUSCs and aging retina-related targets were obtained from GeneCards. Then, Gene Ontology, Kyoto Encyclopedia of Genes and Genomes enrichment analysis, and protein-protein interaction network were analysis, and the expressional level of hub gene was validated by q-PCR. Supernatant addition of hUSCs promoted markedly cellular proliferation, improved viability and inhibited senescence and apoptosis in vitro. A total of 1476 hUSCs-related targets (Relevance score > 20), 692 retinal disease-related targets, and 732 targets related to disease of aging were selected from GeneCards database, and 289 common targets of hUSCs against aging retina were confirmed through Venn analysis. Enrichment analysis demonstrated that hUSCs might exert its anti-apoptosis efficacy in multiple biological processes, including oxidative stress, inflammation and apoptosis, and core targets were associated with HIF-1, MAPK and PI3K-Akt signal. hUSCs inhibited retinal senescence by regulating multiply targets and signaling pathways, of these, HIF-1, MAPK, and PI3K may be important candidates.

4.
Sci China Life Sci ; 67(2): 332-344, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-37480470

RESUMO

Macular corneal dystrophy (MCD) is a progressive, bilateral stromal dystrophic disease that arises from mutations in carbohydrate sulfotransferase 6 (CHST6). Corneal transplantation is the ultimate therapeutic solution for MCD patients. Unfortunately, postoperative recurrence remains a significant challenge. We conducted a retrospective review of a clinical cohort comprising 102 MCD patients with 124 eyes that underwent either penetrating keratoplasty (PKP) or deep anterior lamellar keratoplasty (DALK). Our results revealed that the recurrence rate was nearly three times higher in the DALK group (39.13%, 9/23 eyes) compared with the PKP group (10.89%, 11/101 eyes), suggesting that surgical replacement of the corneal endothelium for treating MCD is advisable to prevent postoperative recurrence. Our experimental data confirmed the robust mRNA and protein expression of CHST6 in human corneal endothelium and the rodent homolog CHST5 in mouse endothelium. Selective knockdown of wild-type Chst5 in mouse corneal endothelium (ACsiChst5), but not in the corneal stroma, induced experimental MCD with similar extracellular matrix synthesis impairments and corneal thinning as observed in MCD patients. Mice carrying Chst5 point mutation also recapitulated clinical phenotypes of MCD, along with corneal endothelial abnormalities. Intracameral injection of wild-type Chst5 rescued the corneal impairments in ACsiChst5 mice and retarded the disease progression in Chst5 mutant mice. Overall, our study provides new mechanistic insights and therapeutic approaches for MCD treatment by high-lighting the role of corneal endothelium in MCD development.


Assuntos
Distrofias Hereditárias da Córnea , Endotélio Corneano , Humanos , Animais , Camundongos , Distrofias Hereditárias da Córnea/genética , Carboidrato Sulfotransferases , Progressão da Doença
5.
Food Chem ; 439: 138095, 2024 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-38039616

RESUMO

Excess formaldehyde (FA) is a strong carcinogen, so the development of a rapid visualized and portable formaldehyde detection platform is of great research importance. A multi-color fluorescence sensing system constituted of model compound (NAHN) and red-emitting InP/ZnS QDs was constructed herein, which can simultaneously realize fluorometric-colorimetric dual-mode sensing when exposed to FA environment. Its preparation process was simplified, the detection process was green, and the limits of detection (LOD) were 0.623 µM and 0.791 µM, respectively. The high recoveries of FA in actual water samples indicated that the sensor had broad application prospects. The prepared fluorescent film can be utilized for rapid visual simulation analysis of FA on the surface of various fruits and vegetables. In addition, a serial logic gate was designed to quickly semi-quantitatively assess FA concentration, which promoted the realization of on-site intelligent evaluation of FA.


Assuntos
Colorimetria , Corantes Fluorescentes , Fluorometria , Formaldeído , Limite de Detecção
6.
ACS Nano ; 17(23): 24055-24069, 2023 Dec 12.
Artigo em Inglês | MEDLINE | ID: mdl-38044579

RESUMO

Hydrogels have attracted tremendous attention as favorable corneal substitutes for treating severe infectious keratitis (IK). However, current hydrogel-based corneal substitutes were majorly designed to promote the single stage of corneal regeneration, which falls short in meeting the clinical management needs of severe IK including the multiple phases of corneal wound healing. Herein, we introduce a versatile hybrid hydrogel (SQPV) composed of silk fibroin and chitosan, which exhibits spatiotemporal properties for drug release. The SQPV is fabricated by incorporating verteporfin-loaded poly(lactic-co-glycolic)-polyethylene glycol-o-nitrobenzene micelles into a hydrogel network, which is formed from methacrylate silk fibroin and glycidyl methacrylate functionalized quaternized chitosan containing polydeoxyribonucleotide. This double network approach results in a material with exceptional anti-inflammatory, antibacterial, and proliferative stimulation and tissue remodeling regulation capabilities. Furthermore, SQPV showcases mechanical strength and transparency akin to those of native cornea. Extensive in vitro and in vivo studies validate SQPV's ability to effectively eliminate residual bacteria, mitigate inflammation, foster regeneration of corneal epithelium and stroma, prevent corneal scarring, and ultimately expedite wound healing. In summary, the SF/CS-based hybrid hydrogel may represent a promising substitute for comprehensive corneal repair and regeneration in severe IK.


Assuntos
Quitosana , Fibroínas , Ceratite , Humanos , Hidrogéis/farmacologia , Quitosana/farmacologia , Liberação Controlada de Fármacos , Cicatrização , Ceratite/tratamento farmacológico , Antibacterianos/farmacologia
7.
Am J Ophthalmol ; 256: 146-155, 2023 12.
Artigo em Inglês | MEDLINE | ID: mdl-37567431

RESUMO

PURPOSE: To investigate the hereditability of corneal tomographic and biomechanical parameters in keratoconus (KC). DESIGN: Prospective cohort study. METHODS: This study was conducted at Qingdao Eye Hospital of Shandong First Medical University in Qingdao, China. Forty-four patients with KC and their biological parents (n = 88) were recruited as the study group. The control group consisted of 84 healthy adults with matched age and gender. Both eyes of each participant underwent clinical examinations, and 1 eye was selected for statistical analysis. Exclusion criteria were as follows: individuals with glaucoma, ocular surgery, systemic diseases known to affect the eyes, or poor cooperation during examination. Subjects were asked to discontinue soft contact lens (CL) wear for 2 weeks and rigid gas permeable CL wear for 4 weeks before ocular examination. All participants underwent a comprehensive assessment including Pentacam Scheimpflug tomography, Corvis ST, visual acuity, refraction examination, axial length, and slitlamp examination for both eyes. Individuals presenting with KC manifestations in at least 1 eye were classified as having KC. A total of 9 Pentacam indices including keratometry in the flat/steep meridian (K1/K2), maximal keratometry (Kmax), thinnest point pachymetry (TP), and maximum/average Ambrósio relational thickness (ARTmax/ARTave), anterior and posterior surfaces elevation of the cornea (Ef/Eb) and total deviation value (Final D), and 21 biomechanical indices were collected. Associations of these factors with KC were evaluated using multiple comparison and binary logistics regression analyses. RESULTS: Two parents (2.27%) from 2 different families were diagnosed with KC. Parents of patients with KC had thinner corneas with altered corneal biomechanical parameters compared with healthy controls (P < .05). The combined tomographic and biomechanical index demonstrated the highest discriminatory power (area under the receiver operating characteristic curve 0.785) and strong specificity (84.5%). Parental corneal tomographic and biomechanical index, Corvis biomechanical index, and TP were identified as the major influential factors for KC in their offspring by logistic regression analysis, with a 73.3% accuracy in identifying offspring with KC. CONCLUSIONS: Parental corneal tomographic and biomechanical properties of patients with KC suggest a possible predisposition to KC. A combination of tomography and corneal biomechanics can be helpful in predicting the incidence rate of KC in the offspring of patients with subclinical KC.


Assuntos
Ceratocone , Adulto , Humanos , Ceratocone/diagnóstico , Estudos Prospectivos , Topografia da Córnea/métodos , Paquimetria Corneana/métodos , Córnea , Curva ROC , Tomografia/métodos , Pais , Fenômenos Biomecânicos
8.
Transl Vis Sci Technol ; 12(6): 1, 2023 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-37261381

RESUMO

Purpose: Our objective was to investigate differences in the ocular surface bacterial composition in cataract patients with and without type 2 diabetes (T2D). Methods: Twenty-four diabetic patients with cataracts (group D) and 14 sex- and age-matched patients with age-related cataracts (group N) were recruited for this study. All samples underwent DNA extraction, fragmentation, purification, library construction, and metagenomic sequencing. Results: The overall conjunctival sac bacterial composition was similar between group D and group N, as determined by alpha diversity and beta diversity. Nevertheless, significant differences were observed in the relative abundance of specific bacteria. At the phylum level, group D had a significantly lower abundance of Chlamydiae, Tenericutes, Chloroflexi, Cyanobacteria, Cossaviricota, Chytridiomycota, Artverviricota, Zoopagomycota, Peploviricota, Deinococcus-Thermus, Preplasmiviricota, and Nucleocytoviricota. At the genus level, group D had a significantly lower abundance of Chlamydia, Mycoplasma, Salmonella, Chryseobacterium, Roseovarius, Desulfococcus, Kangiella, Anaerococcus, and Idiomarina but a significantly higher abundance of Parabacteroides, Phocaeicola, and Sphingomonas. Bacteria such as Aquificae, Parabacteroides, Flavobacterium, Austwickia, Aquifex, Tenacibaculum, and Chryseobacterium in group D and Tenericutes, Chlamydiae, Porphyromonas, Mycoplasma, Chlamydia, Kangiella, Idiomarina, Roseovarius, Aliiroseovarius, and Desulfococcus in group N could be used as conjunctival sac biomarkers, according to the linear discriminant analysis effect size. Gene Ontology functional annotation indicated that bacterial catalytic activity, metabolic processes, locomotion, virion, and reproduction were enriched in group D, while immune system processes were enriched in group N. In addition, the top 30 differentially expressed virulence factors (VFs) were all more enriched in group D. Conclusions: The bacterial composition was similar between the two groups. Several bacterial strains that were reported beneficial in gut were decreased, and pathogenic bacteria were increased in T2D. Furthermore, group D had more active bacterial terms and increased VF expression, suggesting that the susceptibility of diabetic patients to infection is closely related to functional changes in the ocular surface flora. Our conjunctival microbiota atlas provides a reference for investigating ocular complications related to diabetes. Translational Relevance: The altered composition and functional profile of the ocular microbial community in diabetic patients offer evidence for the need to prevent infection during cataract surgery.


Assuntos
Catarata , Diabetes Mellitus Tipo 2 , Microbiota , Humanos , Bactérias/genética , Catarata/complicações , Catarata/genética , Túnica Conjuntiva , Diabetes Mellitus Tipo 2/complicações , Diabetes Mellitus Tipo 2/epidemiologia , Diabetes Mellitus Tipo 2/genética , Metagenoma/genética , Microbiota/genética , Masculino , Feminino
9.
Colloids Surf B Biointerfaces ; 228: 113404, 2023 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-37356140

RESUMO

Fluorescent probes with on-site visual detection function have received extensive attention in the detection of chlortetracycline (CTC), which was widely used in aquaculture and animal husbandry. Copper nanoclusters (Cu NCs) with excellent optical properties were prepared using bovine serum albumin (BSA) as a template, and a multicolor fluorescence strategy based on BSA-stabilized Cu NCs (BSA-Cu NCs) for detecting CTC was proposed. BSA-Cu NCs had a red emission at 640 nm. After the addition of CTC, the red emission of BSA-Cu NCs gradually decreased for internal filtering effect, while the green emission of CTC was significantly enhanced under the sensitization of BSA. This simple sensing process can be achieved in real time by directly mixing the target sample with BSA-Cu NCs, and the detection limit (LOD) of the system for CTC was 12.01 nM. Based on this sensing strategy, a fluorescence film sensing detection platform was constructed to achieve ultra-fast detection of CTC within 30 s. This work provided a fluorescent film sensor with the advantages of portability, ultra-fast and low cost, which provided a feasible alternative for on-site ultra-fast screening of CTC.


Assuntos
Clortetraciclina , Nanopartículas Metálicas , Animais , Cobre , Soroalbumina Bovina , Corantes Fluorescentes , Espectrometria de Fluorescência
10.
Invest Ophthalmol Vis Sci ; 64(5): 26, 2023 05 01.
Artigo em Inglês | MEDLINE | ID: mdl-37234000

RESUMO

Purpose: Contact lens wear (CLW) is one of the leading risk factors for Pseudomonas aeruginosa keratitis (PAK). However, the intrinsic factors that contribute to the high susceptibility to keratitis during CLW remain to be elucidated. CLW over an extended period can elevate corneal norepinephrine (NE) concentration. In this study, we investigated the role of NE in promoting PAK. Methods: We constructed an injury-induced PAK model and a CLW-induced PAK model to confirm the impact of NE during corneal infection. Pharmacological blockage of NE and gene knockdown mouse were used to investigate the downstream effector of NE. RNA sequencing was performed to explore the cellular alterations during NE treatment. Non-parametric Mann-Whitney U test or Kruskal-Wallis test were used to ascertain the significance (P < 0.05). Results: Supplementation of NE led to PAK even without artificial corneal injury during CLW. The effect was mediated by the ß2-adrenergic receptor (ß2-AR) in the corneal epithelium. The ß2-AR blockage by the NE antagonist ICI118,551 (ICI) or by deleting of its encoding gene Adrb2 significantly alleviated infection during CLW. Conversely, ß2-AR activation compromised the integrity of the epithelium and significantly increased the cortical plaque marker ezrin. Transcriptome analysis identified that the protective effect of ICI on the keratitis was mediated by dual-specificity phosphatases. Suramin, a Dusp5 antagonist, abrogated the protective effect of ICI. Conclusions: These data reveal a new mechanism by which NE acts as an intrinsic factor that promotes CLW-induced PAK and provide novel therapeutic targets for treating keratitis by targeting NE-ß2-AR.


Assuntos
Lentes de Contato , Ceratite , Infecções por Pseudomonas , Animais , Camundongos , Pseudomonas aeruginosa/fisiologia , Norepinefrina/farmacologia , Ceratite/etiologia , Lentes de Contato/efeitos adversos , Córnea , Infecções por Pseudomonas/etiologia
11.
Food Chem ; 424: 136376, 2023 Oct 30.
Artigo em Inglês | MEDLINE | ID: mdl-37244186

RESUMO

The abuse of tetracycline antibiotics leads to accumulating residues in the human body, seriously affecting human health. Establishing a sensitive, efficient, and reliable method for qualitative and quantitative detection of tetracycline (TC) is necessary. This study integrated silver nanoclusters and europium-based materials into the same nano-detection system to construct a visual and rapid TC sensor with rich fluorescence color changes. The nanosensor has the advantages of a low detection limit (10.5 nM), high detection sensitivity, fast response, and wide linear range (0-30 µM), which can meet the analysis requirements of different types of food samples. In addition, portable devices based on paper and gloves were designed. Through the smartphone's chromaticity acquisition and calculation analysis application (APP), the real-time rapid visual intelligent analysis of TC in the sample can be realized, which guides the intelligent application of multicolor fluorescent nanosensors.


Assuntos
Nanopartículas Metálicas , Dispositivos Eletrônicos Vestíveis , Humanos , Európio/química , Nanopartículas Metálicas/química , Prata , Corantes Fluorescentes/química , Espectrometria de Fluorescência/métodos , Antibacterianos/análise , Tetraciclina/análise , Limite de Detecção
12.
Gene ; 866: 147349, 2023 May 25.
Artigo em Inglês | MEDLINE | ID: mdl-36893874

RESUMO

To survive under harsh environments, embryonic development of Artemia was arrested at the gastrula stage and released as the diapause embryo. Cell cycle and metabolism were highly suppressed in this state of quiescence. However, cellular mechanisms underlying diapause remain largely unclear. In this study, we found that the expression level of a CT10 regulator of kinase-encoding gene (Ar-Crk) in diapause embryos was significantly lower than non-diapause embryos at the early embryogenetic stage of Artemia. Knockdown of Ar-Crk by RNA interference induced formation of diapause embryos, while the control group produced nauplii. Western blot analysis and metabolic assays revealed that the diapause embryos produced by Ar-Crk-knocked-down Artemia had similar characteristics of diapause markers, arrested cell cycle, and suppressed metabolism with those diapause embryos produced by natural oviparous Artemia. Transcriptomic analysis of Artemia embryos revealed knockdown of Ar-Crk induced downregulation of the aurora kinase A (AURKA) signaling pathway, as well as energetic and biomolecular metabolisms. Taken together, we proposed that Ar-Crk is a crucial factor in determining the process of diapause in Artemia. Our results provide insight into the functions of Crk in fundamental regulations such as cellular quiescence.


Assuntos
Artemia , Diapausa , Animais , Artemia/genética , Regulação para Baixo , Diapausa/genética , Divisão Celular , Ciclo Celular , Embrião não Mamífero/metabolismo
13.
Invest Ophthalmol Vis Sci ; 64(3): 20, 2023 03 01.
Artigo em Inglês | MEDLINE | ID: mdl-36897151

RESUMO

Purpose: To investigate the role of the sympathetic nervous system in corneal neovascularization (CNV) and to identify the downstream pathway involved in this regulation. Methods: Three types of CNV models were constructed with C57BL/6J mice, including the alkali burn model, suture model, and basic fibroblast growth factor (bFGF) corneal micropocket model. Subconjunctival injection of the sympathetic neurotransmitter norepinephrine (NE) was administered in these three models. Control mice received injections of water of the same volume. The corneal CNV was detected using slit-lamp microscopy and immunostaining with CD31, and the results were quantified by ImageJ. The expression of ß2-adrenergic receptor (ß2-AR) was stained with mouse corneas and human umbilical vein endothelial cells (HUVECs). Furthermore, the anti-CNV effects of ß2-AR antagonist ICI-118,551 (ICI) were examined with HUVEC tube formation assay and with a bFGF micropocket model. Additionally, partial ß2-AR knockdown mice (Adrb2+/-) were used to establish the bFGF micropocket model, and the corneal CNV size was quantified based on the slit-lamp images and vessel staining. Results: Sympathetic nerves invaded the cornea in the suture CNV model. The NE receptor ß2-AR was highly expressed in corneal epithelium and blood vessels. The addition of NE significantly promoted corneal angiogenesis, whereas ICI effectively inhibited CNV invasion and HUVEC tube formation. Adrb2 knockdown significantly reduced the cornea area occupied by CNV. Conclusions: Our study found that sympathetic nerves grow into the cornea in conjunction with newly formed vessels. The addition of the sympathetic neurotransmitter NE and activation of its downstream receptor ß2-AR promoted CNV. Targeting ß2-AR could potentially be used as an anti-CNV strategy.


Assuntos
Neovascularização da Córnea , Camundongos , Humanos , Animais , Neovascularização da Córnea/metabolismo , Norepinefrina/farmacologia , Camundongos Endogâmicos C57BL , Córnea/metabolismo , Células Endoteliais da Veia Umbilical Humana/metabolismo , Modelos Animais de Doenças
14.
Chromosoma ; 132(2): 89-103, 2023 06.
Artigo em Inglês | MEDLINE | ID: mdl-36939898

RESUMO

Although parthenogenesis is widespread in nature and known to have close relationships with bisexuality, the transitional mechanism is poorly understood. Artemia is an ideal model to address this issue because bisexuality and "contagious" obligate parthenogenesis independently exist in its congeneric members. In the present study, we first performed chromosome spreading and immunofluorescence to compare meiotic processes of Artemia adopting two distinct reproductive ways. The results showed that, unlike conventional meiosis in bisexual Artemia, meiosis II in parthenogenic Artemia is entirely absent and anaphase I is followed by a single mitosis-like equational division. Interspecific comparative transcriptomics showed that two central molecules in homologous recombination (HR), Dmc1 and Rad51, exhibited significantly higher expression in bisexual versus parthenogenetic Artemia. qRT-PCR indicated that the expression of both genes peaked at the early oogenesis and gradually decreased afterward. Knocking-down by RNAi of Dmc1 in unfertilized females of bisexual Artemia resulted in a severe deficiency of homologous chromosome pairing and produced univalents at the middle oogenesis stage, which was similar to that of parthenogenic Artemia, while in contrast, silencing Rad51 led to no significant chromosome morphological change. Our results indicated that Dmc1 is vital for HR in bisexual Artemia, and the deficiency of Dmc1 may be correlated with or even possibly one of core factors in the transition from bisexuality to parthenogenesis.


Assuntos
Artemia , Recombinases , Animais , Feminino , Recombinases/genética , Artemia/genética , Artemia/metabolismo , Bissexualidade , Meiose , Proteínas de Ciclo Celular/genética , Proteínas de Ciclo Celular/metabolismo , Partenogênese/genética , Rad51 Recombinase/genética , Rad51 Recombinase/metabolismo
15.
Biochem J ; 480(5): 385-401, 2023 03 15.
Artigo em Inglês | MEDLINE | ID: mdl-36852878

RESUMO

Doublesex (DSX) proteins are members of the Doublesex/mab-3-related (DMRT) protein family and play crucial roles in sex determination and differentiation among the animal kingdom. In the present study, we identified two Doublesex (Dsx)-like mRNA isoforms in the brine shrimp Artemia franciscana (Kellogg 1906), which are generated by the combination of alternative promoters, alternative splicing and alternative polyadenylation. The two transcripts exhibited sex-biased enrichment, which we termed AfrDsxM and AfrDsxF. They share a common region which encodes an identical N-terminal DNA-binding (DM) domain. RT-qPCR analyses showed that AfrDsxM is dominantly expressed in male Artemia while AfrDsxF is specifically expressed in females. Expression levels of both isoforms increased along with the developmental stages of their respective sexes. RNA interference with dsRNA showed that the knockdown of AfrDsxM in male larvae led to the appearance of female traits including an ovary-like structure in the original male reproductive system and an elevated expression of vitellogenin. However, silencing of AfrDsxF induced no clear phenotypic change in female Artemia. These results indicated that the male AfrDSXM may act as inhibiting regulator upon the default female developmental mode in Artemia. Furthermore, electrophoretic mobility shift assay analyses revealed that the unique DM domain of AfrDSXs can specifically bind to promoter segments of potential downstream target genes like AfrVtg. These data show that AfrDSXs play crucial roles in regulating sexual development in Artemia, and further provide insight into the evolution of sex determination/differentiation in sexual organisms.


Assuntos
Artemia , Isoformas de RNA , Animais , Masculino , Feminino , Artemia/genética , Isoformas de RNA/metabolismo , Processamento Alternativo , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Diferenciação Sexual/genética
16.
Transl Vis Sci Technol ; 12(2): 21, 2023 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-36786745

RESUMO

Purpose: Previously, we found norepinephrine (NE) could affect the corneal epithelial integrity, herein we investigated the feasibility and safety of NE serving as a chemical enhancer to promote corneal penetration of riboflavin during transepithelial corneal crosslinking (CXL). Methods: The dosage of NE that could promote riboflavin diffusion through the healthy epithelial barrier without inducing epithelial damage in C57BL/6 mice was determined. The safety of NE treatment was confirmed by morphological and histological examinations of the whole cornea. The efficacy of NE in promoting riboflavin penetration was verified by slit lamp and scanning electron microscope (SEM), and corneal biomechanical measurement after CXL. To better fit the clinical scenario, increased NE dosage and shortened riboflavin infiltration time were further evaluated. Results: The lowest dosage of NE (1 mg/mL) that facilitated transepithelial riboflavin permeation was 2 µL. No visible corneal structure alteration was observed after NE treatment. SEM indicated dissociation of intercellular junctions among corneal epithelial cells. Homogenous distribution of riboflavin throughout corneal stroma was observed. NE-treated corneas reached comparable biomechanical properties after CXL, including stress-relaxation curve and elastic modulus, with corneas treated with the commercially available transepithelial drug Peschke TE. To better fit the clinical scenario, increasing NE up to 5.5 µL helped riboflavin infiltrate the corneal stroma within 30 minutes. After CXL with 9 mW/cm2 ultraviolet-A (UVA) for 2.5 minutes, the cornea showed significantly enhanced corneal biomechanical properties with undisturbed corneal endothelium. Conclusions: NE serves as an effective enhancer in increasing riboflavin diffusion with limited impairment on corneal epithelium and has great potential for clinical application. Translation Relevance: NE serves as an effective enhancer for riboflavin penetration and clinical transepithelial CXL.


Assuntos
Norepinefrina , Fármacos Fotossensibilizantes , Animais , Camundongos , Norepinefrina/farmacologia , Fármacos Fotossensibilizantes/farmacologia , Raios Ultravioleta , Camundongos Endogâmicos C57BL , Córnea , Riboflavina/farmacologia , Reagentes de Ligações Cruzadas/farmacologia
17.
Med Image Anal ; 83: 102678, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-36403308

RESUMO

Deformable image registration (DIR) can be used to track cardiac motion. Conventional DIR algorithms aim to establish a dense and non-linear correspondence between independent pairs of images. They are, nevertheless, computationally intensive and do not consider temporal dependencies to regulate the estimated motion in a cardiac cycle. In this paper, leveraging deep learning methods, we formulate a novel hierarchical probabilistic model, termed DragNet, for fast and reliable spatio-temporal registration in cine cardiac magnetic resonance (CMR) images and for generating synthetic heart motion sequences. DragNet is a variational inference framework, which takes an image from the sequence in combination with the hidden states of a recurrent neural network (RNN) as inputs to an inference network per time step. As part of this framework, we condition the prior probability of the latent variables on the hidden states of the RNN utilised to capture temporal dependencies. We further condition the posterior of the motion field on a latent variable from hierarchy and features from the moving image. Subsequently, the RNN updates the hidden state variables based on the feature maps of the fixed image and the latent variables. Different from traditional methods, DragNet performs registration on unseen sequences in a forward pass, which significantly expedites the registration process. Besides, DragNet enables generating a large number of realistic synthetic image sequences given only one frame, where the corresponding deformations are also retrieved. The probabilistic framework allows for computing spatio-temporal uncertainties in the estimated motion fields. Our results show that DragNet performance is comparable with state-of-the-art methods in terms of registration accuracy, with the advantage of offering analytical pixel-wise motion uncertainty estimation across a cardiac cycle and being a motion generator. We will make our code publicly available.

18.
Ophthalmol Ther ; 12(2): 925-938, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36574139

RESUMO

INTRODUCTION: This study aimed to evaluate the habitual reading distance among non-myopic children and also myopic children with undercorrection and with full correction. METHODS: This was a population-based cross-sectional study with a total of 2363 children aged 6-8 years who were recruited from the Hong Kong Children Eye Study. Cycloplegic autorefraction, subjective refraction, habitual visual acuity, and best corrected visual acuity were measured. The entire reading process (9 min) was recorded using a hidden video camera placed 5 m away from the reading desk. Reading distances were taken at 6, 7, 8, and 9 min after the child began reading and were measured using a customized computer program developed in MATLAB. The main outcome was the association of habitual reading distances with refraction status. Habitual reading distances of children were documented via video camera footage. RESULTS: The habitual reading distances of undercorrected myopic children (23.37 ± 4.31 cm) were the shortest when compared to non-myopic children (24.20 ± 4.73 cm, P = 0.002) and fully corrected myopic children (24.81 ± 5.21 cm, P < 0.001), while there was no significant difference between the last two children groups (P = 0.17). A shorter reading distance was associated with myopia (OR 1.67; 95% CI 1.11-2.51; P = 0.013) after adjusting for age, sex, height, near work time, outdoor time, and parental myopia. The association of reading distance with myopia did not hold after undercorrected myopic children were excluded (OR 0.97, 95% CI 0.55-1.73; P = 0.92). A shorter reading distance correlated with poorer vision under habitual correction (ß = - 0.003, P < 0.001). CONCLUSION: A shorter reading distance was present among undercorrected myopic children. Myopia undercorrection is not recommended as a strategy for slowing myopic progression.

19.
Int Ophthalmol ; 43(1): 293-304, 2023 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-35859083

RESUMO

PURPOSE: To determine the bacterial spectrum of exogenous endophthalmitis of different origins, namely, posttraumatic, postcataract surgery, filtering bleb-associated, and intravitreal treatment-related endophthalmitis, using the 16S rDNA sequencing method. METHODS: Aqueous humor or vitreous humor samples were collected from 24 endophthalmitis patients. Traditional cultivation and 16S rDNA sequencing were conducted with these samples. Three senile cataract controls and one intraocular irrigating solution were used as sequencing control. RESULTS: Eleven of the 24 samples (45.8%) obtained positive bacterial cultivation, and each sample positive for only one species. The 11 culture-positive species could all be identified in their corresponding sequencing results, but only four strains being the top one pathogen in the sequencing. A total of 567 species were isolated using 16S rDNA sequencing, with the top five species being Pseudomonas sp., Staphylococcus epidermidis, Staphylococcus sp., Streptococcus sp., and Enterococcus faecalis. The dominant bacterial strains varied among the different endophthalmitis categories but with no significant difference in the overall bacterial spectrum. Bacterial atlas containing Pseudomonas, Streptococcus, Staphylococcus, Actinomycetales_unclassified, Thermus, and Janibacter was shared by the four categories. Aqueous humor bacterial profile showed a higher overlap with contaminating bacteria from the environment. CONCLUSIONS: 16S rDNA sequencing is more efficient for endophthalmitis pathogen screening than the traditional cultivation method in terms of positive detection rate and the number of bacteria identified. But the risk of environmental contamination exists when using 16S rDNA sequencing method for endophthalmitis diagnosis. Different categories of endophthalmitis displayed diversified bacterial composition.


Assuntos
Endoftalmite , Infecções Oculares Bacterianas , Humanos , DNA Ribossômico/genética , Endoftalmite/diagnóstico , Endoftalmite/microbiologia , Bactérias/genética , Humor Aquoso/microbiologia , Infecções Oculares Bacterianas/diagnóstico , Infecções Oculares Bacterianas/microbiologia , DNA Bacteriano/genética
20.
Spectrochim Acta A Mol Biomol Spectrosc ; 284: 121777, 2023 Jan 05.
Artigo em Inglês | MEDLINE | ID: mdl-36058171

RESUMO

A novel multicolor fluorescent nano-probe based on the hybridization of Tb3+ ion with gold nanoclusters (Au NCs) was synthesized to monitor and on-site visual assay of 2,6-pyridinedicarboxylic acid (DPA), a biomarker of bacterial spores. DPA can replace the water molecule in the center of Tb3+ and strongly coordinate with Tb3+ based on the analyte-triggered antenna effect. Simultaneously, the red fluorescence of Au NCs is not influenced after addition of DPA and can be used as steady inside fluorescence reference channel to measure background noise. On this basis, the multicolor fluorescence nano-probe based on Tb3+-doped Au NCs for fast analysis of DPA was fabricated. The linear range of this method is 0 to 12.5 µM and the limit of detection is 3.4 nM, which is well below the quantity of DPA concentration of 60 µM released by the spore transmission dose of anthrax infection. The proposed multicolor fluorescence nano-probe was successfully detecting DPA in actual sample with good sensitivity and specificity. In addition, the visual paper-based nano-probe is designed to detect DPA by using the color scanning application of smart phone. This developed platform possesses abroad application prospects with advantages of effective, convenient carrying, simple operation, good selectivity and repeatability.


Assuntos
Ouro , Térbio , Biomarcadores/análise , Corantes Fluorescentes/análise , Ácidos Picolínicos/análise , Água
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