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Porcine circoviruses disease (PCVD), caused by porcine circovirus (PCVs), is an important swine disease characterized by porcine dermatitis, nephrotic syndrome and reproductive disorders in sows. However, diseases caused by PCV2, PCV3, or PCV4 are difficult to distinguish, so a simple, rapid, accurate and high-throughput diagnostic and identification method is urgently needed to differentiate these three types. In this study, specific primers and probes were designed based on the conserved region sequences of the Rep gene of PCV2, and the Cap gene of PCV3 and PCV4. A multiplex qPCR assay was developed and optimized that the limit of detection concentration could reach as low as 3.8 copies/µL, with all correlation coefficients (R2) exceeding 0.999. Furthermore, the method showed no cross-reaction with other crucial porcine viral pathogens, and both intra-repeatability and inter-reproducibility coefficients of variation were below 2%. The assay was applied to the detection of 738 pig samples collected from 2020 to 2021 in Guangdong Province, China. This revealed positive infection rates of 65.18% for PCV2, 29.27% for PCV3, and 0% for PCV4, with a PCV2/PCV3 co-infection rate of 23.17%. Subsequently, complete genome sequences of 17 PCV2 and 4 PCV3 strains were obtained from the above positive samples and pre-preserved positive circovirus samples. Nucleotide sequence analysis revealed that the 17 PCV2 strains shared 96.7-100% complete nucleotide identity, with 6 strains being PCV2b and 11 strains being PCV2d; the 4 PCV3 strains shared 98.9-99.4% complete nucleotide identity, with 2 strains being PCV3a-1 and 2 strains being PCV3b. This research provides a reliable tool for rapid PCVs identification and detection. Molecular epidemiological investigation of PCVs in pigs in Guangdong Province will help us to understand PCV2 and PCV3 epidemiological characteristics and evolutionary trends.
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Phosphorus (P) and metal accumulation in manured agricultural soils and subsequent losses to waterways have been extensively studied; however, the magnitudes and the factors governing their losses during spring snowmelt flooding are less known. We examined the P and metal release from long-term manured soil to floodwater under simulated snowmelt flooding with recent manure additions. Intact soil columns collected from field plots located in Randolph, Southern Manitoba, 2 weeks after liquid swine manure treatments (surface-applied, injected, or control with no recent manure addition) were flooded and incubated for 8 weeks at 4 ± 1°C to simulate snowmelt conditions. Floodwater (syringe filtered through 0.45 µm) and soil porewater (extracted using Rhizon-Mom samplers) samples were periodically extracted and analyzed for dissolved reactive phosphorus (DRP), pH, zinc (Zn), manganese (Mn), iron (Fe), magnesium (Mg), calcium (Ca), and arsenic (As). Mean floodwater DRP concentrations (mg L-1) for manure injected (2.0 ± 0.26), surface-applied (2.6 ± 0.26), and control (2.2 ± 0.26) treatments did not differ significantly. Despite manure application, DRP loss to floodwater did not significantly increase compared to the control, possibly due to the elevated residual soil P at this site from the long-term manure use. At the end of simulated flooding, the DRP concentrations increased by 1.5-fold and 5-fold in porewater and floodwater, respectively. Metal(loid) concentrations were not affected by manure treatments in general, except for Zn and Mg on certain days. Unlike DRP, where porewater and floodwater concentrations increased with time, metalloid concentration in porewater and floodwater did not show consistent trends with flooding time.
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Inundaciones , Estiércol , Fósforo , Contaminantes del Suelo , Suelo , Contaminantes Químicos del Agua , Fósforo/análisis , Estiércol/análisis , Suelo/química , Contaminantes del Suelo/análisis , Contaminantes Químicos del Agua/análisis , Monitoreo del Ambiente , Metales/análisis , Manitoba , Nieve/químicaRESUMEN
Returning organic nutrient sources (for example, straw and manure) to rice fields is inevitable for coupling crop-livestock production. However, an accurate estimate of net carbon (C) emissions and strategies to mitigate the abundant methane (CH4) emission from rice fields supplied with organic sources remain unclear. Here, using machine learning and a global dataset, we scaled the field findings up to worldwide rice fields to reconcile rice yields and net C emissions. An optimal organic nitrogen (N) management was developed considering total N input, type of organic N source and organic N proportion. A combination of optimal organic N management with intermittent flooding achieved a 21% reduction in net global warming potential and a 9% rise in global rice production compared with the business-as-usual scenario. Our study provides a solution for recycling organic N sources towards a more productive, carbon-neutral and sustainable rice-livestock production system on a global scale.
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Nitrógeno , Oryza , Animales , Nitrógeno/análisis , Agricultura , Suelo , Carbono , Agua , Óxido Nitroso/análisis , Fertilizantes/análisis , GanadoRESUMEN
African swine fever virus (ASFV) has caused enormous economic losses since its first reported detection, and there is still no effective vaccines or drug treatment. During infection, viruses may employ various strategies, such as regulating the host endoplasmic reticulum stress/unfolded protein response or the formation of stress granules (SGs), to form an optimal environment for virus replication. However, how ASFV infection regulates host endoplasmic reticulum stress, eIF2α-regulated protein synthesis, and the formation of SGs remains unclear. Here, we evaluated the activation of ER stress and its three downstream axes during ASFV infection and identified a powerful dephosphorylation of eIF2α by ASFV ex vivo. This strong dephosphorylation property could maintain the efficiency of eIF2α-mediated de novo global protein synthesis, thus ensuring efficient viral protein synthesis at early stage. In addition, the powerful dephosphorylation of eIF2α by ASFV upon infection could also inhibit the formation of SGs induced by sodium arsenite. In addition, a specific eIF2α dephosphorylation inhibitor, salubrinal, could partially counteract ASFV-mediated eIF2α dephosphorylation and inhibit viral replication. Our results provide new insights into the areas of ASFV`s escape from host immunity and hijacking of the host protein translation system.
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Virus de la Fiebre Porcina Africana , Fiebre Porcina Africana , Enfermedades de los Porcinos , Animales , Porcinos , Virus de la Fiebre Porcina Africana/genética , Gránulos de Estrés , Replicación Viral , Biosíntesis de ProteínasRESUMEN
The effects of increased nitrogen (N) deposition on desert ecosystems have been extensively studied from a plant community perspective. However, the response of soil microbial communities, which play a crucial role in nutrient cycling, to N inputs and plant community types remains poorly understood. In this study, we conducted a two-year N-addition experiment with five gradients (0, 10, 30, 60, and 120 kg N ha-1 year-1) to evaluate the effect of increased N deposition on soil bacterial and fungal communities in three plant community types, namely, Alhagi sparsifolia Shap., Karelinia caspia (Pall.) Less. monocultures and their mixed community in a desert steppe located on the southern edge of the Taklimakan Desert, Northwest China. Our results indicate that N deposition and plant community types exerted an independent and significant influence on the soil microbial community. Bacterial α-diversity and community dissimilarity showed a unimodal pattern with peaks at 30 and 60 kg N ha-1 year-1, respectively. By contrast, fungal α-diversity and community dissimilarity did not vary significantly with increased N inputs. Furthermore, plant community type significantly altered microbial community dissimilarity. The Mantel test and redundancy analysis indicated that soil pH and total and inorganic N (NH4+ and NO3-) levels were the most critical factors regulating soil microbial communities. Similar to the patterns observed in taxonomic composition, fungi exhibit stronger resistance to N addition compared to bacteria in terms of their functionality. Overall, our findings suggest that the response of soil microbial communities to N deposition is domain-specific and independent of desert plant community diversity, and the bacterial community has a critical threshold under N enrichment in arid deserts.
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Organic farming in extensive production regions, such as the Canadian prairies have a particularly difficult challenge of replenishing soil reserves of phosphorus (P). Organic grains are exported off the farm while resupply of lost P is difficult due to limited availability of animal manures and low solubility of rock organic fertilizers. As a result, many organic farms on the prairies are deficient in plant-available P, leading to productivity breakdown. A portion of the solution may involve crop genetic improvement. A hypothetical 'catch and release' wheat ideotype for organic production systems is proposed to (i) enhance P uptake and use efficiency but (ii) translocate less P from the vegetative biomass into the grain. Root traits that would improve P uptake efficiency from less-available P pools under organic production are explored. The need to understand and classify 'phosphorus use efficiency' using appropriate indices for organic production is considered, as well as the appropriate efficiency indices for use if genetically selecting for the proposed ideotype. The implications for low seed P and high vegetative P are considered from a crop physiology, environmental, and human nutrition standpoint; considerations that are imperative for future feasibility of the ideotype.
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Leucemia Mieloide Aguda , Linfohistiocitosis Hemofagocítica , Humanos , Linfohistiocitosis Hemofagocítica/diagnóstico , Linfohistiocitosis Hemofagocítica/etiología , Subunidad alfa 2 del Factor de Unión al Sitio Principal/genética , Leucemia Mieloide Aguda/diagnóstico , Leucemia Mieloide Aguda/genética , Translocación Genética , Proteína 1 Compañera de Translocación de RUNX1/genéticaRESUMEN
Objective To prepare specific mouse monoclonal antibody (mAb) against human adenovirus type 55 Hexon protein (HAdV55 Hexon). Methods The Hexon genes of HAdV55, 3, 4, 7, 16 and 21 were chemically synthesized as templates for PCR amplification. The prokaryotic expression plasmids pET28a-HAdV55 Hexon and eukaryotic expression plasmids pCAGGS-HAdV3, 4, 7, 16, 21 and 55 Hexon were constructed respectively. The pET28a-HAdV55 Hexon plasmid was transformed into E. coli competent cell BL21 (DE3) and was induced by IPTG. After the purified inclusion body was denatured and renatured, Hexon55 protein was purified by tangential flow filtration system. pCAGGS-HAdV55 Hexon was used to immunize BALB/c mice by cupping, and HAdV55 Hexon protein was used to booster immunization. The anti-HAdV55 Hexon mAb was prepared by hybridoma technique and the titer and subclass were determined. The specificity of antibody was identified by Western blot using HEK293T cells transfected with pCAGGS-HAdV55 Hexon and by immunofluorescence assay (IFA) using BHK cells transfected with pCAGGS-HAdV55 Hexon. Both clones with high titer were selected, and the cross-reactivity of pCAGGS-HAdV3, 4, 7, 16, 21 and 55 Hexon transfected cells were analyzed by Western blot analysis and IFA. Results PET28a-HAdV55 Hexon and pCAGGS-HAdV55 Hexon, 3, 4, 7, 16 and 21 expression plasmids were successfully constructed. BL21 transformed with pET28a-HAdV55 Hexon was induced by IPTG. The HAdV55 Hexon protein was mainly expressed in the form of inclusion body. After denaturation and renaturation, the purified HAdV55 Hexon protein was obtained by ultrafiltration. Six hybridoma cell lines secreting HAdV55 Hexon mAb were obtained. The antibody subclass analysis showed that 2 strains were IgG2a subtypes and 4 strains were IgG2b. Two specific HAdV55 Hexon antibodies with high titer were obtained, and there was no cross-reactivity with HAdV3, 4, 7, 16, 21 Hexon. Conclusion The specific mice mAb against HAdV55 Hexon provides an experimental basis for establishing its antigen detection method.
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Adenovirus Humanos , Animales , Ratones , Humanos , Adenovirus Humanos/genética , Escherichia coli/genética , Células HEK293 , Isopropil Tiogalactósido , Western Blotting , Inmunoglobulina G , Anticuerpos Monoclonales , Especificidad de Anticuerpos , Ratones Endogámicos BALB CAsunto(s)
Osteomielitis , Leucemia-Linfoma Linfoblástico de Células Precursoras , Humanos , Proteínas de Unión al ADN/genética , Fusión Génica , Proteínas de Fusión Oncogénica/genética , Osteomielitis/genética , Leucemia-Linfoma Linfoblástico de Células Precursoras/genética , Transactivadores/genética , Factores de Transcripción/genética , Femenino , NiñoRESUMEN
Application of nitrification inhibitors (NIs) in agricultural systems is an important strategy to enhance fertilizer nitrogen use efficiency and mitigate soil nitrous oxide (N2O) emissions. Here, we conducted a global meta-analysis of 88 published studies to assess the response of N2O-related functional gene and transcript abundances, and community structure to NIs application. Application of NIs significantly reduced the abundance of ammonia-oxidizing bacteria ammonia monooxygenase (AOB amoA) genes, AOB amoA transcript and nitrite reductase (nirS and nirK) genes. The effectiveness of NIs on reducing the AOB amoA abundance was influenced by N form, soil texture, soil pH and the experimental type (field vs. laboratory). Specifically, NIs were more effective when a mixed inorganic and organic N source was applied to a medium-textured soils. The NIs effectiveness increased with increasing soil pH. The response of AOB amoA abundance to NIs application was not affected by NI type, N rate, soil moisture, soil temperature and soil organic carbon (SOC). The inhibitory effect of NIs on nirS abundance increased with increasing soil temperature. NIs decreased soil nitrifying enzyme activity (NEA) and denitrifying enzyme activity (DEA) by 34.5 % and 27.0 %, respectively, leading to an overall 63.6 % reduction of N2O emissions. Soil NEA correlated positively with the abundance and community structure of AOB amoA but not with AOA amoA. Decrease in DEA with NIs application coincided with the decreasing nirS and nirK abundances. This global-scale assessment demonstrates that the effectiveness of NIs in reducing N2O emissions was attributed to the inhibiting effects on AOB amoA, nirS and nirK genes. Our findings highlight that NIs' inhibition effects on bacterial ammonia-oxidizing community and the encode enzymes in transformation of nitrite to nitric oxide are the main mechanisms for mitigation of N fertilizer-induced N2O emissions.
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Archaea , Suelo , Suelo/química , Nitrificación , Amoníaco/química , Carbono , Fertilizantes/análisis , Microbiología del Suelo , Óxido Nitroso/análisisAsunto(s)
Infecciones por VIH , Micosis , Niño , Humanos , Infecciones por VIH/complicaciones , Micosis/complicacionesRESUMEN
Objective: T cells represent a predominant cell type in autoimmune disease. However, their exact roles are not fully clear in systemic sclerosis (SSc). This study aimed to mainly investigate the alteration in the absolute numbers of T-lymphocyte subsets and the serum levels of cytokines in SSc patients. Methods: A total of 76 patients with SSc and 76 age- and sex-matched healthy controls (HCs) were enrolled. The levels of circulating T cell subsets and serum cytokines were measured by flow cytometry. T cell subsets or serum cytokines correlations with disease activity and organ involvement were analyzed. Results: The absolute numbers of Th2 and Treg cells in SSc patients were lower than those in HCs (p < 0.05), resulting in the ratios of Th1/Th2 [25.01 (12.24, 38.61) vs. 11.64 (6.38, 20.34)] and Th17/Treg [0.42 (0.17, 0.66) vs. 0.17 (0.13, 0.29)] were increased significantly (p < 0.001). The absolute numbers of total T, Th, and Treg cells were negatively correlated with CRP (r = -0.406, p = 0.002; r = -0.263, p < 0.05; r = -0.367 p < 0.01). The serum levels of IL-2, SIL-2R, IL-6, IL-10, INF-γ, and TNF-α were significantly higher than those in HCs (p < 0.001). Increasing IL-2 in the wake of the augment of ESR (r = 0.671, p = 0.004), so did IL-6 (r = 0.378, p < 0.05). The ratio of Th17/Treg in SSc-ILD patients had lower levels than that in other patients [0.35 (0.14, 0.53) vs. 0.64 (0.26, 0.93) p = 0.028]; Treg cells were lessened in patients with Raynaud's phenomenon relative to controls [3.00 (2.41, 4.28) vs. 3.55 (2.86, 4.53) p < 0.05]. The levels of IL-2, IL-10 and INF-γ [3.32 (1.05,11.73) vs. 2.32 (0.44,6.45), p = 0.045], [8.08 (3.63, 355,77) vs. 4.89 (0.78, 21.44), p = 0.02], [6.31 (2.66, 44.03) vs. 4.03 (0.22, 16.96), p = 0.009] were elevated in patients with arthralgia, while the level of Th17 was decreased [0.62 (0.20,2.16) vs. 1.26 (0.22,10.93), p = 0.026]. ROC curve analysis yielded an optimal cut-off IL-2, IL-10, and INF-γ levels of 2.67, 5.93, and 5.32 pg/ml for the presence of arthralgia. Conclusion: We exhibited abnormalities in T subsets and the production of their cytokines in SSc, as compared with those in HCs. This may allow the pathogenesis of SSc and the development of novel therapeutic interventions aimed at targeting these cells and the cytokines they produce.
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Over-application of fertilizers could not improve crop yield and agronomic efficiency, but result in increasing nitrogen (N) surplus and adverse effects on the ecosystem sustainability. Although some previous studies have addressed one or a few environmental aspects in crop production, an integrated assessment for the effects of N fertilizer on multiple environmental impacts, and the optional steps of normalization and weighting is required. A consecutive 2-year plot-based field experiment was conducted with five N fertilizer levels (0, 90, 180, 270, and 360 kg N ha-1) in maize production at three sites in Southwest China, to evaluate the environmental performance and sustainability through joint use of life cycle assessment (LCA) and energy consumption analysis. Results demonstrated that the optimal N rate (180 kg N ha-1) showed greater potential for maintaining high yield (achieved 86% of the yield potential) and reducing the global warming (- 31%), acidification (- 47%), eutrophication (- 44%) compared to farmers' practice, and energy depletion potentials, by reducing pollutants emission during the production and transportation of N fertilizer and Nr losses at farm stage. Optimal N treatment indirectly reduced the land use, life-cycle human toxicity, aquatic eco-toxicity, and terrestrial eco-toxicity potentials by improving grain yield and agronomic efficiency. In addition, the optimal N treatment reduced the energy consumption by enhancing the energy use efficiency (EUE) (+ 74%) and reducing non-renewable energy form (- 45%) than the farmer's practice. This study will provide comprehensive information for both scientists and farmers involved in maize production and N management in subtropical region.
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Contaminantes Ambientales , Fertilizantes , Agricultura/métodos , Efectos Antropogénicos , China , Ecosistema , Contaminantes Ambientales/análisis , Fertilizantes/análisis , Humanos , Nitrógeno/análisis , Zea maysRESUMEN
BACKGROUND: Pain relief for patients in the intensive care unit (ICU) can improve treatment outcomes and reduce the burden on doctors and nurses. This study aims to report the clinical analgesic and sedative effects of nalbuphine and sufentanil on ICU patients. METHODS: This study retrospectively analyzed the medical records of 87 critically ill patients who received nalbuphine or sufentanil infusion in the ICU, including demographic data, diagnosis, Acute Physiology and Chronic Health Evaluation (APACHE) II, Critical Care Pain Observation Tool (CPOT), Richmond Agitation-Sedation Scale (RASS), systolic and diastolic blood pressure, heart rate and blood oxygen saturation (SpO2). The primary outcomes of this study were CPOT and RASS scores. The secondary outcomes were hemodynamic changes, including systolic blood pressure, diastolic blood pressure, heart rate, and SpO2. The adverse events recorded during pain management, such as hypoxemia, respiration depression and bradycardia, were also collected and analyzed. RESULTS: None of the patients in both groups experienced episode of hypoxemia, respiration depression and bradycardia. However, age-stratified analyses showed that nalbuphine has a better analgesic effect than sufentanil for patients aged ≤ 60 (P < 0.05). In contrast, sufentanil showed a better analgesic effect than nalbuphine for patients aged > 60 ( P < 0.05). Furthermore, nalbuphine has a significantly better sedative effect than sufentanil for patients aged ≤ 60 (P < 0.05). CONCLUSION: ICU patients of different age groups may be suitable for different analgesics. For patients under the age of 60, nalbuphine has better analgesia and sedation than sufentanil, and does not cause respiratory depression and drastic hemodynamic changes.
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Nalbufina , Sufentanilo , Anciano , Humanos , Unidades de Cuidados Intensivos , Nalbufina/uso terapéutico , Manejo del Dolor , Estudios Retrospectivos , Sufentanilo/farmacología , Sufentanilo/uso terapéuticoRESUMEN
In the present study, we found the expression of miR-15a-5p (miR-15a) was increased in glioma tissues, and we further explore the underlying mechanism of miR-15a in glioma progression. Microarray analysis used to identify the differentially expressed microRNAs (miRNAs) in glioma tissues. The expression of miR-15a in glioma tissues and cell lines was tested by qRT-PCR. Luciferase assay was used to determine the binding between miR-15a and Smad7. Wound healing and transwell assay were used to examine the role of miR-15a/Smad7 in SHG139 cells. Western blot was used to detect the protein level of Smad7 and epithelial-mesenchymal transition (EMT) markers. A tumor formation model in nude mice was established to measure the role of miR-15a in vivo. MiR-15a was significantly increased in glioma tissues and cells, which indicated a poor prognosis of glioma patients. MiR-15a mimics induced miR-15a level in SHG139 cells, and promoted the malignancy of SHG139 cells, while miR-15a inhibitor showed the opposite effects. Luciferase assay indicated that Smad7 was the direct target of miR-15a, and Smad7 was down-regulated in glioma tissues. Functional experiments revealed that miR-15a inhibitor inhibited the EMT pathway and the migration and invasion of glioma cells, but the silencing of Smad7 reversed the effects of miR-15a inhibitor in EMT pathway and glioma progression. Finally, we performed animal experiments to verify the role of miR-15a in vivo. Present study showed that deletion of miR-15a inhibited the activation of EMT signaling via targeting Smad7, thus suppressed the tumorigenesis and tumor growth of glioma.
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Transición Epitelial-Mesenquimal/genética , Glioma , MicroARNs/genética , Proteína smad7/genética , Animales , Línea Celular Tumoral , Silenciador del Gen , Glioma/genética , Glioma/metabolismo , Glioma/patología , Humanos , Masculino , Ratones , Ratones DesnudosRESUMEN
OBJECTIVE: To analyze the clinical features and genetic variants of two patients from a pedigree affected with Smith-Lemli-Opitz syndrome and explore their genotype-phenotype correlation. METHODS: Clinical data and family history of the pedigree were collected. Whole exome sequencing was carried out to identify the potential variants. Suspected variants were verified by Sanger sequencing of the family members. RESULTS: The proband and her sister both presented with feeding difficulty, facial dysmorphism, seizures, and mental and speech retardation. The third child of this family presented with feeding difficulty, poor weight gain and severe malnutrition after birth. He had died of unknown cause at 6 months without genetic testing. The fourth child was a healthy boy. Genetic testing showed that both the proband and her sister have carried c.127G>T (p.Val43Phe) and c.820_825del (p.Asn274_Val275del) compound heterozygous variants of the DHCR7 gene (NM_001360.2), but the fourth child carried neither of the variants. The two variants were unreported in the literature and disease-related databases, and were not included in the 1000G and gnomAD databases. The c.820_825del variant may affect the sterol-sensitive region of the DHCR7 protein, which can lead to deletion of two amino acids at positions 247 and 275, causing truncation of the DHCR7 protein. It is speculated that this may affect the stability of protein's spatial conformation, thereby decrease the activity of the enzyme. The c.127G>T variant may affect the first transmembrane region of the protein, which is involved in the transmembrane transport of proteins. Multiple software predicted it to be harmful. Conservation analysis suggested that the three amino acids all locate in a highly conserved region of the protein. In consideration of the clinical phenotype, family history and result of genetic testing, we speculated that both patients had Smith-Lemli-Opitz syndrome due to variants of the DHCR7 gene. CONCLUSION: This pedigree has enriched the phenotypic and genotypic data of Smith-Lemli-Opitz syndrome, which clarified the genetic etiology of the patients and provided a basis for genetic counseling of this pedigree.
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Oxidorreductasas actuantes sobre Donantes de Grupo CH-CH , Síndrome de Smith-Lemli-Opitz , China , Femenino , Pruebas Genéticas , Humanos , Masculino , Mutación , Oxidorreductasas actuantes sobre Donantes de Grupo CH-CH/genética , Linaje , Síndrome de Smith-Lemli-Opitz/genéticaRESUMEN
MUC4, a transmembrane mucin, plays important roles in epithelial renewal and differentiation. Recent studies suggest that MUC4 has been implicated in pancreatic cancer pathogenesis and is expressed in various normal and cancer tissues. The underlying features of MUC4 across various cancer types may allow us to ensure appropriate treatment and patient monitoring. However, the contributions of MUC4 to pan-cancer have not been well characterized. In this study, we investigated the expression pattern and prognostic value of MUC4 across multiple databases. We further explored genomic and epigenetic alterations of MUC4, its association with proliferation and metastasis, and the correlation with immune infiltration in different cancers. Our results characterized the distinct expression profile and prognostic values of MUC4 in pan-cancer. Through examining its association with genomic alteration, tumor proliferation, and metastasis, as well as tumor infiltration, we revealed multiple function effects of MUC4. MUC4 may influence prognosis, proliferation, metastasis, and immune response in opposite directions. In conclusion, our findings suggested the necessity to more carefully evaluate MUC4 as a biomarker and therapeutic target and develop the new antibodies for cancer detection and intervention.
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Glycosidases are widely used in food and pharmaceutical industries due to its ability to hydrolyze the glycosidic bonds of various sugar-containing compounds including glycosides, oligosaccharides and polysaccharides to generate derivatives with important physiological and pharmacological activity. While glycosidases often need to be used under high temperature to improve reaction efficiency and reduce contamination, most glycosidases are mesophilic enzymes with low activity under industrial production conditions. It is therefore critical to improve the thermo-stability of glycosidases. This review summarizes the recent advances achieved in engineering the thermo-stability of glycosidases using strategies such as directed evolution, rational design and semi-rational design. We also compared the pros and cons of various techniques and discussed the future prospects in this area.
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Glicósido Hidrolasas , Ingeniería de Proteínas , Glicósido Hidrolasas/genética , Oligosacáridos , PolisacáridosRESUMEN
pH regulation is a serious concern in the industrial fermentation process as pH adjustment heavily utilizes acid/base and pollutes the environment. Under pH-stress conditions, microbial growth and production of valuable target products may be severely affected. Furthermore, some strains generating acidic or alkaline products require self pH regulation and increased tolerance against pH-stress. For pH control, synthetic biology has provided advanced engineering approaches to construct robust and more intelligent microbial strains, exhibiting tolerance to pH-stress to cope with limitations of pH regulation. This study reviewed the current progress of advanced strain evolution strategies to engineer pH-stress tolerant strains via synthetic biology. In addition, a large number of pH-responsive elements, including promoters, riboswitches, and some proteins have been investigated and applied for construction of pH-responsive genetic circuits and intelligent pH-responsive microbial strains.