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Infrared attenuated total reflection spectroscopic analysis and quantitative detection of forage spectral features in ruminant systems
摘要： This study aimed to (1) access protein molecular structure profile and metabolic characteristics of model forages [Foreign sourced-origin (coded as: “FSO”, n = 7 vs. Chinese sourced-origin alfalfa hay “CSO”, n = 5] in ruminant systems; (2) Quantify the relationship between forage protein molecular structures and protein utilization and availability. Advanced non-invasive vibrational molecular spectroscopic technique (ATR-FTIR: Attenuated Total Reflection Fourier Transform Infrared spectroscopy) with chemometrics was applied to reveal forage protein molecular structure. Both univariate and multivariate molecular spectral analyses were applied to study molecular structure features in model forages. The molecular structure study provided the detailed protein structure profiles of Amide I and Amide II areas and height, total Amide I and II area ratios, Amide I to II height ratio as well as Amide I to II area ratio using ATR-FTIR spectroscopy. The results showed FSO and CSO had similar (P > 0.05) protein rumen degradation kinetics. However, FSO had superior quality than CSO in intestinal (IDP) and total digestible protein (TDP) and truly absorbed nutrient supply (P < 0.05). As intestinal digestion of protein, FSO was higher (P < 0.05) in protein digestion in terms of: intestinal digestibility of rumen undegraded protein (dIDP: 47.5 vs. 38.3 %RUP); Intestinal digestible protein (IDP: 17.6 vs. 13.7 %CP). As truly absorbed nutrient supply, FSO contained higher (P < 0.05) truly absorbed rumen synthesized microbial protein, absorbable rumen undegradable feed protein in the small intestine, total truly digested protein in the small intestine, metabolizable protein and Feed Milk Value (FMVDVE: 1.2 vs. 1.1 g/kg DM). The molecular structure-nutrition interactive relationship study showed that protein molecular structure profiles were highly associated to protein rumen degradation kinetics, significantly correlated to protein subfractions, protein intestinal digestion, and truly absorbed nutrient supply in ruminant systems.
关键词： Vibrational molecular spectroscopy-ATR-FTIR,Intestine digestion,Model forages,Truly absorbed protein supply,Rumen degradation,Protein molecular structure and conformation,Univariate and multivariate molecular spectral analyses
更新于2025-09-12 10:27:22
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Accuracy and biases in predicting the chemical and physical traits of many types of cheeses using different visible and near-infrared spectroscopic techniques and spectrum intervals
摘要： Near-infrared spectroscopy (NIRS) has been widely used to determine various composition traits of many dairy products in the industry. In the last few years, near-infrared (NIR) instruments have become more and more accessible, and now, portable devices can be easily used in the field, allowing the direct measurement of important quality traits. However, the comparison of the predictive performances of different NIR instruments is not simple, and the literature is lacking. These instruments may use different wavelength intervals and calibration procedures, making it difficult to establish whether differences are due to the spectral interval, the chemometric approach, or the instrument’s technology. Hence, the aims of this study were (1) to evaluate the prediction accuracy of chemical contents (5 traits), pH, texture (2 traits), and color (5 traits) of 37 categories of cheese; (2) to compare 3 instruments [2 benchtop, working in reflectance (R) and transmittance (T) mode (NIRS-R and NIRS-T, respectively) and 1 portable device (VisNIRS-R)], using their entire spectral ranges (1100–2498, 850–1048, and 350–1830 nm, respectively, for NIRS-R, NIRS-T and VisNIRS-R); (3) to examine different wavelength intervals of the spectrum within instrument, comparing also the common intervals among the 3 instruments; and (4) to determine the presence of bias in predicted traits for specific cheese categories. A Bayesian approach was used to develop 8 calibration models for each of 13 traits. This study confirmed that NIR spectroscopy can be used to predict the chemical composition of a large number of different cheeses, whereas pH and texture traits were poorly predicted. Color showed variable predictability, according to the trait considered, the instrument used, and, within instrument, according to the wavelength intervals. The predictive performance of the VisNIRS-R portable device was generally better than the 2 laboratory NIRS instruments, whether with the entire spectrum or selected intervals. The VisNIRS-R was found suitable for analyzing chemical composition in real time, without the need for sample uptake and processing. Our results also indicated that instrument technology is much more important than the NIR spectral range for accurate prediction equations, but the visible range is useful when predicting color traits, other than lightness. Specifically for certain categories (i.e., caprine, moldy, and fresh cheeses), dedicated calibrations seem to be needed to obtain unbiased and more accurate results.
关键词： chemometric,texture trait,cheese quality,water-soluble protein,Bayesian calibration
更新于2025-09-12 10:27:22
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Development of Highly Reliable SERS‐active Photonic Crystal Fiber Probe and its Application in the Detection of Ovarian Cancer Biomarker in Cyst Fluid
摘要： Conventionally Surface enhanced Raman spectroscopy (SERS) is realized by adsorbing analytes onto nano-roughened planar substrate coated with noble metals (silver or gold) or their colloidal nanoparticles (NPs). Nano scale irregularities in such substrates/NPs could lead to SERS sensors with poor reproducibility and repeatability. Herein, we demonstrate a suspended core photonic crystal fiber (PCF) based SERS sensor with extremely high reproducibility and repeatability in measurement with a relative standard deviation of only 1.5% and 4.6% respectively, which makes it more reliable than any existing SERS sensor platforms. In addition, our platform could improve the detection sensitivity owing to the increased interaction area between the guided light and the analyte, which is incorporated into the holes that runs along the length of the PCF. Numerical calculation established the significance of the interplay between light coupling efficiency and evanescent field distribution, which could eventually determine the sensitivity and reliability of the developed SERS active-PCF sensor. As a proof of concept, using this sensor, we demonstrated the detection of haptoglobin, a biomarker for ovarian cancer, contained within the ovarian cyst fluid, which facilitated in differentiating the stages of the cancer. We envision that with necessary refinements, this platform could potentially be translated as a next generation highly sensitive SERS-active opto-fluidic biopsy needle for the detection of biomarkers in body fluids.
关键词： reproducibility and repeatability in measurement,Surface enhanced Raman scattering,photonic crystal fiber,biopsy needle,protein sensing,ovarian cancer
更新于2025-09-12 10:27:22
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Semiconductor Quantum Dots Are Efficient and Recyclable Photocatalysts for Aqueous PET-RAFT Polymerization
摘要： This Letter describes the use of CdSe quantum dots (QDs) as photocatalysts for photoinduced electron transfer reversible addition?fragmentation chain transfer (PET-RAFT) polymerization of a series of aqueous acrylamides and acrylates. The high colloidal solubility and photostability of these QDs allowed polymerization to occur with high efficiency (>90% conversion in 2.5 h), low dispersity (PDI < 1.1), and ultralow catalyst loading (<0.5 ppm). The use of protein concentrators enabled the removal of the photocatalyst from the polymer and monomer with tolerable metal contamination (8.41 ug/g). These isolated QDs could be recycled for four separate polymerizations without a significant decrease in efficiency. By changing the pore size of the protein concentrators, the QDs and polymer could be separated from the remaining monomer, allowing for the synthesis of block copolymers using a single batch of QDs with minimal purification steps and demonstrating the fidelity of chain ends.
关键词： block copolymers,CdSe quantum dots,photocatalysts,aqueous acrylamides,acrylates,protein concentrators,PET-RAFT polymerization
更新于2025-09-12 10:27:22
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Matrix-assisted pulsed laser evaporation of β-glucosidase from a dopa/quinone target
摘要： β-glucosidase (BG) plays a key role in determining the efficiency of the enzymatic complex cellulase for the degradation of cellulose into sugars. It hydrolyses the cellobiose, an inhibitor of the enzymatic complex. Therefore, the immobilization of BG is a great challenge for the industrial application of cellulases. Cellulases usually contains a BG amount insufficient to avoid inhibition by cellobiose. Here the BG was immobilized by matrix assisted pulsed laser evaporation (MAPLE) technique. The frozen matrix was composed of water, water/m-DOPA and water/m-DOPA/quinone. The effect of the excipients on the final conformation of the enzyme after the MAPLE processing was determined. The enzyme secondary structure was studied by FTIR analysis. The catalytic performances of the deposited films were tested in the cellobiose hydrolysis reaction. The results demonstrate that the presence of the oxidized form of m-DOPA, the O-quinone form, can protect the protein native structure, with the laser inducing little or no damage. In fact, only the samples deposited from this target preserved the secondary structure of the polypeptide chain and allowed a complete hydrolysis of cellobiose for four consecutive runs, showing a high operational stability of the biocatalyst.
关键词： MAPLE,β-glucosidase,Thin films,Protein conformation,m-DOPA,O-quinone
更新于2025-09-12 10:27:22
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Role of Clathrin Protein to Increase Electric Current on Dye-Sensitized Solar Cell (DSSC) Based Natural Dyes
摘要： Dye-Sensitized Solar Cell (DSSC) has the cost of fabrication is relatively cheap, easy to produce, work on the area of the visible light and environmentally friendly. The shortage of DSSC, namely the efficiency is still low compared with solar cells of silicon. In this study, have been successfully fabricated DSSC with clathrin protein deposited. The method used in this research is to deposit clathrin protein in porous TiO2 semiconductors with a concentration of 0%, 25%, 50%, and 75%. Tests carried out on the DSSC that has been made current testing electricity using a solar simulator to determine the changes in electrical current that occur in DSSC. The conclusion from this research is that with increasing concentration of the addition of the clathrin protein in the DSSC leads to an increase in the electric current generated by DSSC. From the results of testing using a solar simulator obtained the value of electric current is the highest, namely 5.247 mA with the addition of the concentration of the clathrin protein by 75%.
关键词： electric current,Dye-Sensitized Solar Cell,TiO2,clathrin protein,DSSC
更新于2025-09-12 10:27:22
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Step-growth polymerization of traptavidin-DNA conjugates for plasmonic nanostructures
摘要： Here, we use two important biomaterials, protein and DNA, to construct self-assembled nanostructures through Watson-Crick base-paring of DNAs. We apply a simple magnetic separation method to purify traptavidin-DNA conjugates, and demonstrate synthesis of linear arrays of traptavidin-DNA conjugates via the step-growth polymerization approach with pre-determined DNA sequences. Using the traptavidin-DNA array as a template, we assemble gold nanoparticles to form linear plasmonic nanostructures in a programmable manner. The traptavidin-DNA conjugates thus provide a convenient platform for one-dimensional assembly of biotinylated nanomaterials for many biomedical applications from drug delivery to bio-sensing.
关键词： Traptavidin,Plasmonic nanostructure,Supramolecular assembly,Protein-DNA conjugate,DNA nanostructure
更新于2025-09-12 10:27:22
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Replacing Pumps with Light Controlled Insulin Delivery
摘要： Purpose of Review The aim of this review is to summarize the development of the photoactivated depot (PAD) approach for the minimally invasive and continuously variable delivery of insulin. Recent Findings Using an insulin PAD, we have demonstrated that we can release native, bioactive insulin into diabetic animals in response to light signals from a small external LED light source. We have further shown that this released insulin retains bioactivity and reduces blood glucose. In addition, we have designed and constructed second generation materials that have high insulin densities, with the potential for multiple day delivery. Summary The PAD approach for insulin therapy holds promise for addressing the pressing need for continuously variable delivery methods that do not rely on pumps, and their myriad associated problems.
关键词： Depot,Glucagon,Insulin,Artificial pancreas,Protein delivery,Insulin pump,Light
更新于2025-09-12 10:27:22
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The in vivo mechanics of the magnetotactic backbone as revealed by correlative FLIM-FRET and STED microscopy
摘要： Protein interaction and protein imaging strongly benefit from the advancements in time-resolved and superresolution fluorescence microscopic techniques. However, the techniques were typically applied separately and ex vivo because of technical challenges and the absence of suitable fluorescent protein pairs. Here, we show correlative in vivo fluorescence lifetime imaging microscopy F?rster resonance energy transfer (fLiM-fRet) and stimulated emission depletion (SteD) microscopy to unravel protein mechanics and structure in living cells. We use magnetotactic bacteria as a model system where two proteins, MamJ and MamK, are used to assemble magnetic particles called magnetosomes. The filament polymerizes out of MamK and the magnetosomes are connected via the linker MamJ. Our system reveals that bacterial filamentous structures are more fragile than the connection of biomineralized particles to this filament. More importantly, we anticipate the technique to find wide applicability for the study and quantification of biological processes in living cells and at high resolution.
关键词： FLIM-FRET,living cells,magnetotactic bacteria,STED microscopy,protein mechanics
更新于2025-09-12 10:27:22
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A Robust and General Approach to Quantitatively Conjugate Enzymes to Plasmonic Nanoparticles
摘要： Bioconjugates of plasmonic nanoparticles have received considerable attention due to their potential biomedical applications. Succesfull bioconjugation requires control over the number and activity of the conjugated proteins, and the colloidal stability of the particles. In practice, this requires re-optimization of the conjugation protocol for each combination of protein and nanoparticle. Here we report a robust and general protocol that allows for the conjugation of a range of proteins to di?erent types of nanoparticles using very short polyethylene-glycol(PEG) linkers, while simultaneously preserving protein activity and colloidal stability. The use of short linkers ensures that the protein is located close to the particle surface, where their refractive index sensitivity and near-?eld enhancement is maximal. We demonstrate that the use a Tween20 containing stabilizing bu?er is critical in maintaining colloidal stability and protein function throughout the protocol. We obtain quantitative control over the average number of enzymes per particle by either varying the number of functional groups on the particle, or the enzyme concentration during incubation. This new route of preparing quantitative protein-nanoparticle bioconjugates paves the way to develop rational and quantitative strategies to functionalize nanoparticles for applications in sensing, medical diagnostics and drug delivery.
关键词： medical diagnostics,drug delivery,quantitative control,protein activity,plasmonic nanoparticles,colloidal stability,sensing,bioconjugation,PEG linkers,Tween20
更新于2025-09-12 10:27:22