Current Research in Biotechnology最新文献

Smart colorimetric sensor for the naked-eye detection of food freshness is considered as the most attractive tool in food safety. Herein, we prepared curcumin (CUR) coated electrospun regenerated cellulose nanofiber (RCA) composites (CUR-Cot), to monitor the real-time spoilage of raw chicken. The physicochemical properties of the CUR-Cot sensor were completely studied. The surface modification, morphology, crystalline nature, and thermal stability of CUR-Cot were investigated by using various spectral, analytical, and microscopic techniques. Based on the results, the successful coating of CUR on the surface of RCA was confirmed. Interestingly, the CUR-Cot showed a significant change in total color difference value (ΔE, 0 days − 0.0–14.93, after 1 day – 14.93–23.64, after 2 days – 23.64–44.78, after 3 days- 44.78–55.22, and after 4 days – 55.22–60.96, detectable by the naked eye) in the real-time monitoring for chicken freshness. In addition, the present CUR-Cot smart colorimetric sensor is reversible with change in pH, and the reversed sensor can be reused. Furthermore, the hydrophobic nature of the CUR-Cot was confirmed by water contact angle analysis (WCA, contact angle of 90 ± 3.00), which increases its application and adaptability. Good antibacterial, barrier, and optical properties of the present CUR-Cot were also found. Overall, the results confirm that the usage of CUR-Cot sensor can be highly efficient, user-friendly, and non-destructive for the real-time monitoring of chicken freshness.

Background

Head and neck squamous cell carcinoma (HNSCC) is a common invasive malignant tumor that lacks powerful predictive or prognostic biomarkers. Ferroptosis and cuproptosis are two new forms of programmed cell death. Our study was aimed at constructing a prognostic model with a combination of cuproptosis and ferroptosis-related genes (CFRGs) for the early clinical detection of HNSCC.

Methods

We obtained the information of CFRGs, including the RNASeq data and corresponding clinical data in HNSCC patients from the TCGA and GEO databases. We assessed 28 CFRGs, and analyzed the relationship between those genes and their clinical features and prognosis of HNSCC. The consensus cluster analysis was employed to generate three CFRGclusters. Then, we investigated the association of molecular patterns and prognostic significance in these subtypes. The clinical indicators of the prognosis-related genes were identified and prognostic CFRG_score were constructed. We then built a predictive nomogram with confirmed consistency and reliability by calibration curve analysis. At last, we verified the expression of CFRGs in HNSCC tissues by qRT-PCR and immunohistochemical results.

Results

The DEGs were different between the normal and HNSCC tumor tissues and we screened out 28 CFRGs related to the prognosis in HNSCC. Associations between the clinical information and prognosis were found in the molecular subtypes related to prognosis. We utilized enrichment analysis of the differential genes and showed that those DEGs were mostly enriched in the biological processes associated with the pathways of neurodegeneration-multiple diseases, Alzheimer disease, Prion disease, Parkinson disease and Amyotrophic lateral sclerosis. CFRG_score was established to predict the survival of HNSCC patients and found that higher CFRG_score suggested favorable OS for patients, indicating the prediction of better prognosis. Moreover, we created highly reliable nomogram which could predict well for the expected prognosis. In addition, we confirmed that the expression of EGFR, VEGFA, HSPA5, SLC3A2, CAV1 and CD44 were consistent with qRT-PCR and immunohistochemical analysis in HNSCC tissues by qRT-PCR.

Conclusions

This prognostic model based on prognostic differential CFRG_score is strongly related to clinical characteristics, prognosis, and therapy in HNSCC patients and could be used as a promising tool which is dedicated to guiding the treatment of HNSCC.

In vitro studies of stone-related crystals and crystals-cell interactions have been extensively done to investigate cellular, molecular and pathogenic mechanisms leading to renal calculi. Effective preparation of various types of stone-related crystals is thus crucial for such studies. Nevertheless, various protocols for preparing these stone-related crystals were scatteredly reported without comparative analysis of their efficacies and yields. Herein, we systematically compared our protocols (with the suffix “-Si”) for preparing calcium oxalate (CaOx) monohydrate (COM), CaOx dihydrate (COD), magnesium ammonium phosphate (struvite), uric acid (UA), calcium phosphate dihydrate (brushite), hydroxyapatite (HAP), and calcium carbonate (CaCO₃) crystals with other protocols published previously. The morphological evaluation revealed that our protocols provided the most homogeneous and most typical monoclinic prismatic, bipyramidal, coffin lid and rectangle shapes of COM, COD, struvite and UA crystals, respectively. There were comparable morphological results for brushite, HAP and CaCO₃ crystals generated by different protocols. Our protocols provided the greatest yield for generating brushite crystals but with lower yields for others. Chemical analysis by Fourier transform infrared (FTIR) spectroscopy revealed comparable results among different protocols to generate each crystal type. In summary, all these protocols can be used to generate each crystal type. But our protocols offer the best quality, in terms of homogeneity and typical shape, for generating COM, COD, struvite and UA crystals.

Severe inflammation in joints caused by the detrimental effects of the immune system is termed Rheumatoid arthritis. The unconstrained proliferation of immune cells and pro-inflammatory cytokines deteriorates Synovium which secretes synovial fluid to lubricate joints and cartilage. Non-steroidal anti-inflammatory drugs (NSAIDs) are the only therapeutics for treating rheumatoid arthritis, and long-term intake causes serious side effects on the organs. Fucoidan, a sulfated polysaccharide found on the cell walls of brown algae shows bioactive potential. In our study, fucoidan was extracted from Padina pavonica (PD), Stoechospermum marginatum (StM), Spatolossum macrodontum (SpM), Dictyota bartayresiana (DD), and Turbinaria decurrens (TD) and evaluated for anti-inflammatory and anti-arthritis activities. Fucoidan was extracted and evaluated for anti-inflammatory activity in vitro using RAW 264.7 macrophage cell lines, followed by in vivo anti-arthritis activity on Wistar male rats. Nitric oxide suppression was comparatively high in fucoidan from TD (IC₅₀ − 12.93 µg/mL). Purified fucoidan from TD, significantly reduced inflammation, size of paw edema, downregulated proinflammatory cytokines (IL-6, IL-1β, TNF-α), and upregulated anti-inflammatory cytokine (IL10) in CFA-induced arthritis in Wistar male rats. Biochemical parameters like SOD, CAT, GSH, GPX, and GST and haematological parameters like total-protein, albumin, haemoglobin, and RBC were upregulated, and other parameters like urea, uric acid, creatinine, bilirubin, SGOT, SGPT, ALP, WBC, ESR, RF, and CRP were downregulated. Histopathology of the liver, kidney, and ankle joints reveals that fucoidan intake restrained inflammation and tissue damage. Therefore, fucoidan extracted from TD is a potential candidate for the treatment of rheumatoid arthritis.

Creatine kinase-MM (CK-MM) is a relatively muscle-specific enzyme with a plasma half-life of approximately 2 h. Total creatine kinase elevation is evident in several conditions associated with acute muscle injury, severe muscular exertion, and myocardial infarction. The presence of a large amount of the CK-MM enzyme in blood serum is a biomarker of muscular injuries and cardiac assault. In this study, we developed a structure-switching aptamer that can be immobilized on a sensor to detect CK-MM. CK-BB was used as a counter-target to ensure the specific targeting of CK-MM. Melting-Off SELEX was employed to develop aptamers exhibiting significant structural changes on binding. The selected aptamer shows a high affinity towards CK-MM with a K_d value of 14.7 nM.

In the last years, many research efforts have been applied for the development of filamentous fungi as hosts for heterologous protein production. Aspergillus vadensis CBS 113365, a close relative of the industrial workhorse Aspergillus niger, has been suggested as a more suitable cell factory as it does not acidify the culture medium and produces very low levels of secreted proteases. Therefore, efficient methods and tools that allow the genetic manipulation and exploitation of this biotechnologically relevant fungus are needed. To date, only protoplast-mediated transformation and classical cloning strategies have been implemented for A. vadensis genetic modification, which decreases the exploitation capacity of this fungus at the industrial level. In this study, we have adapted and implemented an Agrobacterium tumefaciens-mediated transformation protocol for A. vadensis for the first time, and applied the FungalBraid system to genetically modify this species by means of synthetic biology. As proof of concept, we have successfully complemented and fluorescently labelled a uridine auxotrophic A. vadensis pyrA^- strain and generated A. vadensis mutants carrying the Penicillium expansum-based expression cassette for the heterologous production of the antifungal protein PeAfpA from P. expansum. Even though we have yet to find the conditions that trigger PeAfpA production in this species, the implementation of the ATMT method reported here, along with the application of the FungalBraid system, will greatly aid in this task and will facilitate the exploitation of A. vadensis as a fungal workhorse for protein production for multiple biotechnological applications.

Light quality (spectral arrangement) and quantity (photoperiod and intensity) influence plant growth and metabolism and also interact with several factors including environmental parameters in defining the plant behavior. The Light Emitting Diode (LED) lights are extensively utilized in the cultivation of several plant species, especially horticultural plants due to their lower power consumption and higher luminous efficiency compared to the conventional fluorescent lights. The aim of this review paper is to examine the potential of LED technology as it relates to plant lighting in greenhouses and other horticultural environments. It also desires to give an in-depth study of the advantages of LED lighting on plant development, yield, the production of secondary metabolites, and defense mechanisms. Horticultural lighting might undergo a revolution because LEDs are used in solid-state lighting, which would be a tremendous advancement after decades of research. LEDs may be used in a variety of horticulture lighting applications, such as tissue culture lighting, controlled environment research lighting, supplementary lighting, and photoperiod lighting for greenhouses. The primary impacts of light colors on plant performance are shown by the spectrum effects of LEDs as an independent source of light, together with the diverse sensitivity of many plant species and alternatives. LED light influences performance of enzyme, gene expression, cell wall formation, plant defense and postharvest quality. The spectrum reactions are mediated by the ambient lighting in a greenhouse, which also indicates a strong relationship between the additional supplementary lighting and changing environmental factors. LEDs are growing further to become cost-effective for even large-scale horticulture lighting applications as light output increases and device expenditures decrease.

Leishmaniasis is recognised as the second largest parasitic disease worldwide and yet a neglected disease. The current pharmacological treatments are associated with significant challenges, including high toxicity, high cost and parasitic resistance. Considering the potential of isopentyl caffeate (ICaf) as an anti-leishmanial agent, the present work evaluated the in vivo toxicity of ICaf and the absorption, distribution, metabolism, and excretion (ADME) properties in silico, aiming at the treatment of Leishmania amazonensis. For the in vivo toxicity testing, Swiss mice (Mus musculus) were treated with a single dose of ICaf. During the 14-day evaluation period, the animals underwent assessments including hippocratic screening, weight measurement, as well as histological and hematological evaluations. Analysis of ADME properties of ICaf was conducted to evaluate its pharmacokinetic characteristics and bioavailability. Characteristics, such as molar refractivity through Lipinski's Rule of Five, were identified. The in silico results showed that ICaf is considered to have good oral bioavailability and has potential to be considered as a new drug. From the in vivo toxicity testing, none of the evaluated parameters revealed toxicity of ICaf to the animals when treated intraperitoneally. The in vivo treatment reduced the lesion and the parasite load at the tested doses, corroborating the assumption that ICaf may be a potential pharmacological alternative against L. amazonensis.