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1.
Commun Biol ; 7(1): 474, 2024 Apr 18.
Artigo em Inglês | MEDLINE | ID: mdl-38637717

RESUMO

Coding transcript-derived siRNAs (ct-siRNAs) produced from specific endogenous loci can suppress the translation of their source genes to balance plant growth and stress response. In this study, we generated Arabidopsis mutants with deficiencies in RNA decay and/or post-transcriptional gene silencing (PTGS) pathways and performed comparative sRNA-seq analysis, revealing that multiple RNA decay and PTGS factors impede the ct-siRNA selective production. Genes that produce ct-siRNAs often show increased or unchanged expression and typically have higher GC content in sequence composition. The growth and development of plants can perturb the dynamic accumulation of ct-siRNAs from different gene loci. Two nitrate reductase genes, NIA1 and NIA2, produce massive amounts of 22-nt ct-siRNAs and are highly expressed in a subtype of mesophyll cells where DCL2 exhibits higher expression relative to DCL4, suggesting a potential role of cell-specific expression of ct-siRNAs. Overall, our findings unveil the multifaceted factors and features involved in the selective production and regulation of ct-siRNAs and enrich our understanding of gene silencing process in plants.


Assuntos
Proteínas de Arabidopsis , Arabidopsis , Arabidopsis/metabolismo , RNA Interferente Pequeno/genética , RNA Interferente Pequeno/metabolismo , Proteínas de Arabidopsis/metabolismo , Interferência de RNA , RNA de Cadeia Dupla/metabolismo , Plantas/metabolismo
2.
Langmuir ; 2024 Apr 16.
Artigo em Inglês | MEDLINE | ID: mdl-38626327

RESUMO

In this work, a trimetallic (Ni/Co/Zn) organic framework (tMOF), synthesized by a solvothermal method, was calcinated at 400 and 600 °C and the final products were used as a support for lipase immobilization. The material annealed at 400 °C (Ni-Co-Zn@400) had an improved surface area (66.01 m2/g) and pore volume (0.194 cm3/g), which showed the highest enzyme loading capacity (301 mg/g) with a specific activity of 0.196 U/mg, and could protect the enzyme against thermal denaturation at 65 °C. The optimal pH and temperature for the lipase were 8.0 and 45 °C but could tolerate pH levels 7.0-8.0 and temperatures 40-60 °C. Moreover, the immobilized enzyme (Ni-Co-Zn@Lipase, Ni-Co-Zn@400@Lipase, or Ni-Co-Zn@600@Lipase) could be recovered and reused for over seven cycles maintaining 80, 90, and 11% of its original activity and maintained a residual activity >90% after 40 storage days. The remarkable thermostability and storage stability of the immobilized lipase suggest that the rigid structure of the support acted as a protective shield against denaturation, while the improved pH tolerance toward the alkaline range indicates a shift in the ionization state attributed to unequal partitioning of hydroxyl and hydrogen ions within the microenvironment of the active site, suggesting that acidic residues may have been involved in forming an enzyme-support bond. The high enzyme loading capacity, specific activity, encouraging stability, and high recoverability of the tMOF@Lipase indicate that a multimetallic MOF could be a better platform for efficient enzyme immobilization.

3.
medRxiv ; 2024 Mar 26.
Artigo em Inglês | MEDLINE | ID: mdl-38585974

RESUMO

Most current studies rely on short-read sequencing to detect somatic structural variation (SV) in cancer genomes. Long-read sequencing offers the advantage of better mappability and long-range phasing, which results in substantial improvements in germline SV detection. However, current long-read SV detection methods do not generalize well to the analysis of somatic SVs in tumor genomes with complex rearrangements, heterogeneity, and aneuploidy. Here, we present Severus: a method for the accurate detection of different types of somatic SVs using a phased breakpoint graph approach. To benchmark various short- and long-read SV detection methods, we sequenced five tumor/normal cell line pairs with Illumina, Nanopore, and PacBio sequencing platforms; on this benchmark Severus showed the highest F1 scores (harmonic mean of the precision and recall) as compared to long-read and short-read methods. We then applied Severus to three clinical cases of pediatric cancer, demonstrating concordance with known genetic findings as well as revealing clinically relevant cryptic rearrangements missed by standard genomic panels.

4.
Nanotechnology ; 35(23)2024 Mar 18.
Artigo em Inglês | MEDLINE | ID: mdl-38497442

RESUMO

In contrast to lithium-ion batteries, lithium-sulfur batteries have higher theoretical energy density and lower cost, so they would become competitive in the practical application. However, the shuttle effect of polysulfides and slow oxidation-reduction kinetics can degrade their electrochemical performance and cycle life. In this work, we have first developed the porous FeNi Prussian blue cubes as precursors. The calcination in different atmospheres was employed to make precursors convert into common pyrolysis products or novel carbon-based phosphides, and sulfides, labeled as FeNiP/A-C, FeNiP/A-P, and FeNiP/A-S. When these products serve as host materials in the sulfur cathode, the electrochemical performance of lithium-sulfur batteries is in the order of S@FeNiP/A-P > S@FeNiP/A-S > S@FeNiP/A-C. Specifically, the initial discharge capacity of S@FeNiP/A-P can reach 679.1 mAh g-1at 1 C, and the capacity would maintain 594.6 mAh g-1after 300 cycles. That is because the combination of carbon-based porous structure and numerous well-dispersed Ni2P/Fe2P active sites contribute FeNiP/A-P to obtain larger lithium-ion diffusion, lower resistance, stronger chemisorption, and more excellent catalytic effect than other samples. This work may deliver that metal-organic framework-derived carbon-based phosphides are more suitable to serve as sulfur hosts than carbon-based sulfides or common pyrolysis products for enhancing Li-S batteries' performance.

5.
J Mol Med (Berl) ; 102(3): 313-335, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38265445

RESUMO

Endothelial cell (EC) dysfunction is associated with atherosclerosis. Circular RNAs (circRNAs) are covalently closed loops formed by back-splicing, are highly expressed in a tissue-specific or cell-specific manner, and regulate ECs mainly through miRNAs (mircoRNAs) or protein sponges. This review describes the regulatory mechanisms and physiological functions of circRNAs, as well as the differential expression of circRNAs in aberrant ECs. This review focuses on their roles in inflammation, proliferation, migration, angiogenesis, apoptosis, senescence, and autophagy in ECs from the perspective of signaling pathways, such as nuclear factor κB (NF-κB), nucleotide-binding domain, leucine-rich-repeat family, pyrin-domain-containing 3 (NLRP3)/caspase-1, Janus kinase/signal transducer and activator of transcription (JAK/STAT), and phosphoinositide-3 kinase/protein kinase B (PI3K/Akt). Finally, we address the issues and recent advances in circRNAs as well as circRNA-mediated regulation of ECs to improve our understanding of the molecular mechanisms underlying the progression of atherosclerosis and provide a reference for studies on circRNAs that regulate EC dysfunction and thus affect atherosclerosis.


Assuntos
Aterosclerose , MicroRNAs , Humanos , RNA Circular , Fosfatidilinositol 3-Quinases/metabolismo , MicroRNAs/genética , MicroRNAs/metabolismo , Transdução de Sinais , Aterosclerose/genética
6.
Int J Biol Macromol ; 254(Pt 3): 127901, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-37952798

RESUMO

This work describes the synthesis of fibrous nickel-based metal organic framework (Ni-ZIF) via simple solvothermal method. The material formed was calcinated at 400, 600, 800 °C to improve its surface area, porosity and enzyme binding capacity. Changes in X-ray diffraction pattern after calcination revealed the Ni-ZIF transitioned from amorphous to crystalline structure. The surface area, pore volume and pore size for Ni-ZIF@600 were found to be 312.15 m2/g, 0.88 cm3/g and 10.28 nm, with an enzyme loading capacity of 593.85 mg/g after 30 h The free (ß-Gal-LEH) and immobilized ß-Galactosidase were stable at pH 7.5, temperature 50 °C, and yielded 70.70 and 63.95 mM glucose after milk lactose hydrolysis, respectively. The Ni-ZIF@600@ß-Gal-LEH exhibited high enzyme retention capacity, maintaining 59.44 % of its original activity after 6-cycles. The enhanced magnetic property, enzyme binding capacity and easy recoverability of the calcinated Ni-ZIF could guarantee its industrial significance as immobilization module for enzyme-mediated catalysis.


Assuntos
Enzimas Imobilizadas , Níquel , Níquel/química , Enzimas Imobilizadas/química , Temperatura , beta-Galactosidase/química , Fenômenos Magnéticos
8.
Heliyon ; 9(11): e21915, 2023 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-38034615

RESUMO

Nkx2.3, a transcription factor, plays important roles in various developmental processes. However, the mechanisms underlying nkx2.3's regulation of pouch and pharyngeal arch development in zebrafish remain unclear. In this study, we demonstrated that knockdown or knockout of nkx2.3 resulted in the absence of posterior ceratobranchial cartilages in zebrafish. The absence of posterior pharyngeal cartilages is a consequence of the compromised proliferation and differentiation and survival of cranial neural crest cells (CNCCs). Notably, we found that nkx2.3 was not involved in endoderm pouch formation. Additionally, our findings suggested that nkx2.3 negatively regulated Fibroblast growth factor (Fgf) signaling, as overexpression of fgf8 could mimic the phenotype observed in nkx2.3 morphants, suppressing CNCC differentiation. Moreover, inhibiting Fgf signaling restored the abnormalities in posterior cartilages induced by nkx2.3 knockdown. These findings establish the essential role of nkx2.3 in the development of posterior ceratobranchial cartilages through the inhibition of fgf8.

9.
Med Sci Monit ; 29: e941937, 2023 Oct 21.
Artigo em Inglês | MEDLINE | ID: mdl-37864329

RESUMO

BACKGROUND Lumbar disc herniation (LDH) at L4-L5 impacts paravertebral muscle morphology. Intervertebral disc degeneration is linked to paravertebral muscle changes, affecting LDH treatment outcomes. This study explored L4-L5 LDH paravertebral muscle alterations, specifically in the erector spinae, multifidus, and psoas major, using Michigan State University's classification to guide LDH treatment. MATERIAL AND METHODS The study enrolled 160 patients, including 39 normal patients and 121 L4-L5 LDH patients. Patients with LDH were grouped according to MSU classification and compared to the normal group according to demographics and imaging changes. RESULTS In patients with L4-L5 herniation in Zone B, the FI of the ES muscle at L3-L4 level, L4-L5 level, and L5-S1 level was higher than that of normal people (P=0.018, P=0.043, P=0.010, respectively), and there was no difference between FI of MF and normal people. The Zone B patients also had a smaller CSA of the ES muscle at L4-L5 level than that in the normal group (P=0.049). Patients in the Zone C group were older than those in the normal group (P=0.014). The CSA of the PM of patients with Grade 3 herniation differed from that of the normal group at the L4-L5 and L5-S1 level. They were higher than in normal people at L4-L5 level (P=0.011) and lower at L5-S1 level (P=0.028). CONCLUSIONS In patients with L4-L5 herniation in Zone B, the FI of ES at L3-S1 level was higher than in normal people, and the CSA at L4-L5 level was smaller than in normal people. In patients with Grade3 herniation, PM CSA was larger at L4-L5 level and smaller at L5-S1 level than in normal people.


Assuntos
Degeneração do Disco Intervertebral , Deslocamento do Disco Intervertebral , Humanos , Michigan , Universidades , Vértebras Lombares , Imageamento por Ressonância Magnética/métodos , Músculos Psoas
10.
Heliyon ; 9(10): e20794, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37860527

RESUMO

As a zoonotic parasite, Cryptosporidium spp. could cause severe diarrhea mainly in calves and children globally. Monitoring and prevention of Cryptosporidium spp.'s prevalence is of great significance in both economy and public health aspects. In this study, specific primers and probes were designed within the conserved region of 18S rRNA gene for Cryptosporidium spp. and recombinase polymerase amplification assays based on the fluorescence monitoring (real-time RPA) as well as combined with a lateral flow strip (LFS RPA) were developed. Both of the two RPA assays allowed the exponential amplification of the target fragment within 20 min. After incubation on a metal bath at 42 °C, the LFS RPA results were displayed on the lateral flow strip within 5 min while real-time RPA allowed the real-time observation of the results in Genie III at 39 °C. The RPA assays showed high specificity for Cryptosporidium spp. without any cross-reaction with other tested pathogens causing diarrhea in cattle. With the recombinant plasmid DNA containing the 18S rRNA gene of Cryptosporidium spp. serving as a template, the limit of detection for real-time RPA and LFS RPA assays were 14.6 and 12.7 copies/reaction, respectively. Moreover, the RPA assays were validated by testing diarrheic cattle fecal samples and compared with a real-time PCR. The positive ratio of Cryptosporidium spp. was 24.04 % (44/183) and 26.23 % (48/183) in both RPA assays and real-time PCR assay, respectively, and the kappa coefficient value was 0.942. The diagnostic specificity and diagnostic sensitivity of both RPA assays were 100 % and 91.67 %, respectively. Forty-one of 48 positive samples were successfully sequenced and four Cryptosporidium species were detected, including C. parvum (n = 20), C. andersoni (n = 17), C. bovis (n = 3) and C. ryanae (n = 1). The developed RPA assays are easy to operate and faster to obtain the detection results, and they are suiting for the point-of-care detection and facilitating the prevention and control of Cryptosporidium spp. infections.

11.
Psych J ; 2023 Oct 31.
Artigo em Inglês | MEDLINE | ID: mdl-37905907

RESUMO

Previous study indicates that there are two distinct behavioral patterns in the sensory-motor synchronization task with short stimulus onset asynchrony (SOA; 2-3 s) or long SOA (beyond 4 s). However, the underlying neural indicators and mechanisms have not been elucidated. The present study applied magnetoencephalography (MEG) technology to examine the functional role of several oscillations (beta, gamma, and mu) in sensorimotor synchronization with different SOAs to identify a reliable neural indicator. During MEG recording, participants underwent a listening task without motor response, a sound-motor synchronization task, and a motor-only continuation task. These tasks were used to explore whether and how the activity of oscillations changes across different behavioral patterns with different tempos. Results showed that during both the listening and the synchronization task, the beta oscillation changes with the tempo. Moreover, the event-related synchronization of beta oscillations was significantly correlated with motor timing during synchronization. In contrast, mu activity only changes with the tempo in the synchronization task, while the gamma activity remains unchanged. In summary, the current study indicates that beta oscillation could be an indicator of behavioral patterns between fast tempo and slow tempo in sensorimotor synchronization. Also, it is likely to be the potential mechanism of maintaining rhythmic continuous movements with short SOA, which is embedded within the 3 s time window.

12.
Sci Total Environ ; 903: 166605, 2023 Dec 10.
Artigo em Inglês | MEDLINE | ID: mdl-37640078

RESUMO

Nitrous acid (HONO) is an established precursor of hydroxyl (OH) radical and has significant impacts on the formation of PM2.5 and O3. Despite extensive research on HONO observation in recent years, knowledge regarding its sources and sinks in urban areas remains inadequate. In this study, we monitored the atmospheric concentrations of HONO and related pollutants, including gaseous nitric acid and particulate nitrate, simultaneously at a supersite in downtown Chengdu, a megacity in southwestern China during spring, when was chosen due to its tolerance for both PM2.5 and O3 pollution. Furthermore, we employed the random forest model to fill the missing data of HONO, which exhibited good predictive performance (R2 = 0.96, RMSE = 0.36 ppbv). During this campaign, the average mixing ratio of HONO was measured to be 1.0 ± 0.7 ppbv. Notably, during periods of high O3 and PM2.5 concentrations, the mixing ratio of HONO was >50 % higher compared to the clean period. We developed a comprehensive parameterization scheme for the HONO budget, and it performed well in simulating diurnal variations of HONO. Based on the HONO budget analysis, we identified different mechanisms that dominate HONO formation at different times of the day. Vehicle emissions and NO2 heterogeneous conversions were found to be the primary sources of HONO during nighttime (21.0 %, 30.2 %, respectively, from 18:00 to 7:00 the next day). In the morning (7:00-12:00), NO2 heterogeneous conversions and the reaction of NO with OH became the main sources (35.0 %, 32.2 %, respectively). However, in the afternoon (12:00-18:00), the heterogeneous photolysis of HNO3 on PM2.5 was identified as the most substantial source of HONO (contributing 52.5 %). This study highlights the significant variations in primary HONO sources throughout the day.

13.
Int J Hyperthermia ; 40(1): 2240548, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37544652

RESUMO

OBJECTIVE: This nonrandomized prospective clinical trial aimed to assess the efficacy, safety and follow-up outcomes of ultrasound-guided high-intensity focused ultrasound (USgHIFU) surgery in patients with breast fibroadenoma. METHODS: With the approval of the institutional ethics committee and written informed consent, a total of 113 patients diagnosed with breast fibroadenoma by core-needle biopsy in our hospital were recruited. USgHIFU surgery was performed under local anesthesia. Contrast-enhanced ultrasound (CEUS) or contrast-enhanced MRI (CEMRI) was performed to evaluate the nonperfused volume (NPV). The patients were followed up with physical examination and ultrasound imaging. RESULTS: The clinical outcome of 85 patients with 147 fibroadenomas with a follow-up time of more than 3 months was analyzed in this study. Fifty-two patients had one lesion, twenty-one patients had two lesions and twelve patients had more than two lesions. During USgHIFU, the median localization time for all fibroadenomas was 3 (interquartile range: 1, 5) min, and the median treatment time was 9 (interquartile range: 5, 15) min. Under local anesthesia, all the patients tolerated the treatment well. No serious epidermal burns were observed in any of the patients. Based on CEUS or CEMRI imaging evaluation, the median NPV ratio was 100% (interquartile range: 79.2%, 116.8%). The VRR were 26.77 ± 50.05%, 50.22 ± 42.01% and 72.74 ± 35.39% at 3-6 months, 6-12 months and >12 months, respectively, which showed significant statistical difference (p < .001). CONCLUSION: Ultrasound-guided HIFU surgery is an effective and safe noninvasive alternative technique for the treatment of breast fibroadenoma.


Assuntos
Neoplasias da Mama , Fibroadenoma , Ablação por Ultrassom Focalizado de Alta Intensidade , Humanos , Feminino , Fibroadenoma/diagnóstico por imagem , Fibroadenoma/cirurgia , Estudos Prospectivos , Estudos de Viabilidade , Ultrassonografia , Neoplasias da Mama/diagnóstico por imagem , Neoplasias da Mama/cirurgia , Ablação por Ultrassom Focalizado de Alta Intensidade/métodos , Ultrassonografia de Intervenção/métodos , Resultado do Tratamento
14.
Genome Res ; 33(7): 1089-1100, 2023 07.
Artigo em Inglês | MEDLINE | ID: mdl-37316351

RESUMO

Recent studies exploring the impact of methylation in tumor evolution suggest that although the methylation status of many of the CpG sites are preserved across distinct lineages, others are altered as the cancer progresses. Because changes in methylation status of a CpG site may be retained in mitosis, they could be used to infer the progression history of a tumor via single-cell lineage tree reconstruction. In this work, we introduce the first principled distance-based computational method, Sgootr, for inferring a tumor's single-cell methylation lineage tree and for jointly identifying lineage-informative CpG sites that harbor changes in methylation status that are retained along the lineage. We apply Sgootr on single-cell bisulfite-treated whole-genome sequencing data of multiregionally sampled tumor cells from nine metastatic colorectal cancer patients, as well as multiregionally sampled single-cell reduced-representation bisulfite sequencing data from a glioblastoma patient. We show that the tumor lineages constructed reveal a simple model underlying tumor progression and metastatic seeding. A comparison of Sgootr against alternative approaches shows that Sgootr can construct lineage trees with fewer migration events and with more in concordance with the sequential-progression model of tumor evolution, with a running time a fraction of that used in prior studies. Lineage-informative CpG sites identified by Sgootr are in inter-CpG island (CGI) regions, as opposed to intra-CGIs, which have been the main regions of interest in genomic methylation-related analyses.


Assuntos
Metilação de DNA , Neoplasias , Humanos , Metilação de DNA/genética , Sulfitos , Análise de Sequência de DNA/métodos , Genoma , Neoplasias/genética , Ilhas de CpG/genética
15.
J Invertebr Pathol ; 198: 107921, 2023 06.
Artigo em Inglês | MEDLINE | ID: mdl-37023892

RESUMO

Nucleic acid sequence-based amplification (NASBA) provides a fast and convenient approach for nucleic acid amplification under isothermal conditions, and its combination with an immunoassay-based lateral flow dipstick (LFD) could produce a higher detection efficiency for M. rosenbergii nodavirus isolated from China (MrNV-chin). In this study, two specific primers and a labelled probe of the capsid protein gene of MrNV-chin were constructed. The process of this assay mainly included a single-step amplification at a temperature of 41 â„ƒ for 90 min, and hybridization with an FITC-labeled probe for 5 min, with the hybridization been required for visual identification during LFD assay. The test results indicated that, the NASBA-LFD assay showed sensitivity for 1.0 fg M. rosenbergii total RNA with MrNV-chin infection, which was 104 times that of the present RT-PCR approach for the detection of MrNV. In addition, no products were created for shrimps with infection of other kinds of either DNA or RNA virus, which indicated that the NASBA-LFD was specific for MrNV. Therefore, the combination of NASBA and LFD is a new alternative detection method for MrNV which is rapid, accurate, sensitive and specific without expensive equipment and specialised personnel. Early detection of this infectious disease among aquatic organisms will help implement efficient therapeutic strategy to prevent its spread, enhance animal health and limit loss of aquatic breeds in the event of an outbreak.


Assuntos
Nodaviridae , Palaemonidae , Vírus de RNA , Animais , Replicação de Sequência Autossustentável , Nodaviridae/genética , Vírus de RNA/genética , Técnicas de Amplificação de Ácido Nucleico/métodos
16.
Nat Commun ; 14(1): 1335, 2023 Mar 11.
Artigo em Inglês | MEDLINE | ID: mdl-36906683

RESUMO

Oceanic eddy-induced meridional heat transport (EHT) is an important process in the Southern Ocean heat budget, the variability of which significantly modulates global meridional overturning circulation (MOC) and Antarctic sea-ice extent. Although it is recognized that mesoscale eddies with scales of ~40-300 km greatly contribute to the EHT, the role of submesoscale eddies with scales of ~1-40 km remains unclear. Here, using two state-of-the-art high-resolution simulations (resolutions of 1/48° and 1/24°), we find that submesoscale eddies significantly enhance the total poleward EHT in the Southern Ocean with an enhancement percentage reaching 19-48% in the Antarctic Circumpolar Current band. By comparing the eddy energy budgets between the two simulations, we detect that the primary role of submesoscale eddies is to strengthen mesoscale eddies (and thus their heat transport capability) through inverse energy cascade rather than directly through submesoscale heat fluxes. Due to the submesoscale-mediated enhancement of mesoscale eddies in the 1/48° simulation, the clockwise upper cell and anti-clockwise lower cell of the residual-mean MOC in the Southern Ocean are weakened and strengthened, respectively. This finding identifies a potential route to improve the mesoscale parameterization in climate models for more accurate simulations of the MOC and sea ice variability in the Southern Ocean.

17.
BMC Vet Res ; 18(1): 339, 2022 Sep 08.
Artigo em Inglês | MEDLINE | ID: mdl-36076203

RESUMO

BACKGROUND: Bovine rotavirus A (BRVA) is considered to be the most common pathogen of severe diarrhea in cattle worldwide, which could lead to the death of newborn calves and cause the significant economic losses to the cattle industry. As a novel isothermal nucleic acid amplification technique, recombinase polymerase amplification (RPA) has been applied widely for the rapid detection of different important pathogens in human and animals. RESULTS: An RT-RPA assay based on the real time fluorescence monitoring (real-time RT-RPA) and an RT-RPA assay combined with a lateral flow strip (LFS RT-RPA) were successfully developed by targeting the VP6 gene of BRVA. The RT-RPA assays allowed the exponential amplification of the target fragment in 20 min. After incubation of the LFS RT-RPA on a metal bath at 40 °C, the results were displayed on the lateral flow strip within 5 min, while real-time RT-RPA allowed the real-time observation of the results in Genie III at 42 °C. Both of the two assays showed high specificity for BRVA without any cross-reaction with the other tested pathogens causing diarrhea in cattle. With the standard RNA of BRVA serving as a template, the limit of detection for real-time RT-RPA and LFS RT-RPA were 1.4 × 102 copies per reaction and 1.4 × 101 copies per reaction, respectively. In the 134 fecal samples collected from cattle with diarrhea, the BRVA positive rate were 45.52% (61/134) and 46.27% (62/134) in real-time RT-RPA and LFS RT-RPA, respectively. Compared to a previously published real-time PCR, the real-time RT-RPA and LFS RT-RPA showed a diagnostic specificity of 100%, diagnostic sensitivity of 98.39% and 100%, and a kappa coefficient of 0.985 and 1.0, respectively. CONCLUSIONS: In this study, BRVA was successfully detected in cattle fecal samples by the developed real-time RT-RPA and LFS RT-RPA assays. The developed RT-RPA assays had great potential for the rapid detection of BRVA in under-equipped diagnostic laboratory and the point-of-need diagnosis at quarantine stations and farms, which is of great importance to control BRVA-associated diarrhea in cattle herds.


Assuntos
Transcrição Reversa , Rotavirus , Animais , Bovinos , Diarreia/diagnóstico , Diarreia/veterinária , Humanos , Técnicas de Amplificação de Ácido Nucleico/métodos , Técnicas de Amplificação de Ácido Nucleico/veterinária , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Recombinases/genética , Rotavirus/genética , Sensibilidade e Especificidade
18.
Front Vet Sci ; 9: 953599, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35990282

RESUMO

Bovine kobuvirus (BKV) is a novel kobuvirus considered to be closely related to calf diarrhea and has become a worldwide epidemic. Currently, the BKV lacks an efficient and convenient detection method to assist the research on BKV prevalence. In this study, a new and specific TaqMan-based real-time RT-PCR for the detection of BKV was developed using the conserved region of the 3D gene. The assay was highly specific for BKV, without cross-amplification with other non-targeted pathogens. The limit of detection of this assay was 102 copies. Standard curves showed a strong linear correlation from 102 to 106 copies of BKV standard RNA per reaction, and the parameters revealed as a slope of -3.54, efficiency of 91.64%, and regression coefficients (R2) of 0.998. The assay was also reproducible, with the intra-assay and inter-assay coefficient of variation <1.0%. The newly developed real-time RT-PCR was validated using 243 fecal samples collected from diarrheic or non-diarrheic cattle from nine regions in Hebei province and revealed the positive detection of BKV at a ratio of 19.34% (47/243). Sequencing of partial 3D genes from 13 positive samples and the following phylogenetic analysis demonstrated the reliability of the assay. In conclusion, the newly developed TaqMan-based real-time RT-PCR could be used for the screening and epidemic monitoring of BKV.

19.
Water Sci Technol ; 85(9): 2639-2651, 2022 May.
Artigo em Inglês | MEDLINE | ID: mdl-35576258

RESUMO

The reduction of soluble U(VI) to insoluble U(IV) by photocatalytic technology is considered to be a valid method to remove U(VI) from water. Herein, g-C3N4/Ag/TiO2 Z-scheme heterojunction was synthesized for photocatalytic U(VI) reduction application. The SEM, XRD and XPS characterization results showed that a ternary g-C3N4/Ag/TiO2 composite photocatalyst was synthesized successfully. g-C3N4/Ag/TiO2 exhibited excellent photocatalytic reduction performance for U(VI) under visible light irradiation. After 30 min irradiation, the removal rate of U(VI) was above 99%. XPS indicated that the majority of U(VI) on the surface of g-C3N4/Ag/TiO2 was reduced to U(IV). In addition, the photocatalytic activity of g-C3N4/Ag/TiO2 has been kept significantly after five rounds of experiments, indicating good stability. g-C3N4/Ag/TiO2 exhibited better photocatalytic reduction of U(VI) under visible light irradiation, which is mainly ascribed to Z-scheme photocatalytic mechanism assisted by the LSPR effect (Local Surface Plasmon Resonance). Ag with plasmon resonance effect on the loading has a strong absorption of photon energy. In addition, an intermediate charge transfer channel is formed between Ag and the semiconductor to inhibit the combination of photogenerated electrons and holes, resulting in a significant increase in the photocatalytic activity of the photocatalyst. This idea has some significance in design of other composite photocatalytic systems.

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