Determination of the Efficiency of Magnetic Resonance Imaging in the Evaluation of Compressive Myelopathy.

Background The phrase "compressive myelopathy" refers to compression of the spinal cord, either internally or externally. This compression might arise from various sources such as a herniated disc, post-traumatic compression, and epidural abscess as well as epidural or intradural neoplasms. Magnetic resonance imaging (MRI) plays a crucial role in differentiating between compressive and non-compressive myelopathy. After eliminating compressive lesions, attention is directed toward intrinsic cord-related causes of acute myelopathy including vascular, infectious, and inflammatory pathologies. Aims The study aimed to assess different etiologies of compressive myelopathy, analyze the MRI features of spinal cord compressive lesions, classify the lesions depending on site, and correlate MRI findings with intraoperative findings and histopathology in operated cases. Material & methods A total of 50 patients, who exhibited clinical symptoms indicative of compressive myelopathy sent to the Radiology department, Rangaraya Medical College (RMC), Kakinada for MRI spine were included in the study. It's an observational cross-sectional study. Statistical Package for Social Sciences (SPSS) version 22.0 (IBM Corp., Armonk, USA) was used for statistical calculations. Result Among the 50 cases of compressive myelopathy, the etiologies are distributed as follows: trauma (22 cases), infection (12 cases), primary neoplasm (eight cases), and secondary neoplasm (eight cases); extradural compressive lesions (84%) and Intradural-extramedullary lesions (16%). Conclusion Utilizing MRI successfully assessed the spinal cord integrity and characterized spinal tumors. Consequently, the study concludes that MRI is a highly definitive, sensitive, and accurate tool for evaluating compressive myelopathy.

Tailoring Synthetic Polypeptide Design for Directed Fibril Superstructure Formation and Enhanced Hydrogel Properties.

The biological significance of self-assembled protein filament networks and their unique mechanical properties have sparked interest in the development of synthetic filament networks that mimic these attributes. Building on the recent advancement of autoaccelerated ring-opening polymerization of amino acid N-carboxyanhydrides (NCAs), this study strategically explores a series of random copolymers comprising multiple amino acids, aiming to elucidate the core principles governing gelation pathways of these purpose-designed copolypeptides. Utilizing glutamate (Glu) as the primary component of copolypeptides, two targeted pathways were pursued: first, achieving a fast fibrillation rate with lower interaction potential using serine (Ser) as a comonomer, facilitating the creation of homogeneous fibril networks; and second, creating more rigid networks of fibril clusters by incorporating alanine (Ala) and valine (Val) as comonomers. The selection of amino acids played a pivotal role in steering both the morphology of fibril superstructures and their assembly kinetics, subsequently determining their potential to form sample-spanning networks. Importantly, the viscoelastic properties of the resulting supramolecular hydrogels can be tailored according to the specific copolypeptide composition through modulations in filament densities and lengths. The findings enhance our understanding of directed self-assembly in high molecular weight synthetic copolypeptides, offering valuable insights for the development of synthetic fibrous networks and biomimetic supramolecular materials with custom-designed properties.

Non-Covalent Assembly of Multiple Fluorophores in Edible Protein/Lipid Hydrogels for Applications in Multi-Step Light Harvesting and White-Light Emission.

The design and production of biodegradable and sustainable non-toxic materials for solar-energy harvesting and conversion is a significant challenge. Here, our goal was to report the preparation of novel protein/lipid hydrogels and demonstrate their utility in two orthogonal fundamental studies-light harvesting and white-light emission. Our hydrogels contained up to 90% water, while also being self-standing and injectable with a syringe. In one application, we loaded these hydrogels with suitable organic donor-acceptor dyes and demonstrated the energy-transfer cascade among four different dyes, with the most red-emitting dye as the energy destination. We hypothesized that the dyes were embedded in the protein/lipid phase away from the water pools as monomeric entities and that the excitation of any of the four dyes resulted in intense emission from the lowest-energy acceptor. In contrast to the energy-transfer cascade, we demonstrate the use of these gels to form a white-light-emitting hydrogel dye assembly, in which excitation migration is severely constrained. By restricting the dye-to-dye energy transfer, the blue, green, and red dyes emit at their respective wavelengths, thereby producing the composite white-light emission. The CIE color coordinates of the emission were 0.336 and 0.339-nearly pure white-light emission. Thus, two related studies with opposite requirements could be accommodated in the same hydrogel, which was made from edible ingredients by a simple method. These gels are biodegradable when released into the environment, sustainable, and may be of interest for energy applications.

Advances in Structural Modifications and Properties of Graphene Quantum Dots for Biomedical Applications.

Graphene quantum dots (GQDs) are carbon-based, zero-dimensional nanomaterials and unique due to their astonishing optical, electronic, chemical, and biological properties. Chemical, photochemical, and biochemical properties of GQDs are intensely being explored for bioimaging, biosensing, and drug delivery. The synthesis of GQDs by top-down and bottom-up approaches, their chemical functionalization, bandgap engineering, and biomedical applications are reviewed here. Current challenges and future perspectives of GQDs are also presented.

Biogenic Photo-Catalyst TiO2 Nanoparticles for Remediation of Environment Pollutants.

This article reports a benign environmentally friendly fabrication method of titanium dioxide (TDO) nanoparticles (named TDO NPs3, TDO NPs5, and TDO NPs8) using aqueous extract of durva herb waste. This synthesis process avoids use of harmful substances and persistent chemicals throughout the order and enables us to control the size of the nanomaterials. Characterization of TDO nanoparticles was analyzed by ultraviolet-visible spectroscopy, X-ray diffraction, and Fourier transform infrared spectroscopy. The morphological nature of the TDO samples was inspected by transmission electron microscopy, which indicated that the TDO NPs3, TDO NPs5, and TDO NPs8 were spherical in shape, with average sizes of 5.14, 12.54, and 29.61 nm, respectively. The stability of TDO nanoparticles was assessed using thermogravimetric analysis and dynamic light scattering analysis. These samples could be used for degradation of polluting industrial textile dyes, such as methylene blue (MB) and rhodamine B (Rh-B). Remarkably, the TDO NPs3 sample (5.14 nm size) exhibits a noticeable degradation of the MB dye in a shorter time period (50 min) than the TDO NPs8 sample with a size of 29.61 nm (120 min). The TDO NPs3 sample was also tested for degradation of Rh-B dye, showing high degradation efficiency over a short period of time (60 min). In contrast, the TDO NPs8 sample showed degradation of the Rh-B dye in 120 min. The effect of the dye concentration and the catalyst dose to remove dye pollutants has also been investigated. The synthesized TDO NPs act as exceptional catalysts for the degradation of dyes, and they are promising materials for the degradation of industrial polluting dyes.

Nanoarmored Multi-Enzyme Cascade Catalysis.

This chapter reports a single-step preparation of nanoarmored bi-enzyme systems assembled on 1-D and 2-D nanomaterials, with glucose oxidase and peroxidase enzymes as model systems for cascade bio-catalysis. This is a simple and facile method to both exfoliate the bulk 1D (carbon nanotubes, CNT) and 2D nanomaterials (α-Zirconium phosphate, α-ZrP) and bind the enzymes in a single step. Exfoliation of the bulk material enhances the accessible surface area of the materials for the enzyme binding, and it also boosts the diffusion of reagents from the bulk phase to the active sites of the bio-catalysts. For example, a mixture of horseradish peroxidase, glucose oxidase, and bovine serum albumin (BSA) were adsorbed on the surfaces of the α-ZrP nanoplates or carbon nanotubes (CNT) as the bulk materials are exfoliated simultaneously, in a one-step process. The resulting bio-catalysts were thoroughly characterized by powder X-ray diffraction, electron microscopy, biochemical and biophysical methods, while enzyme activity studies proved successful binding of enzymes with retention of activities or even enhancements in their specific activities. For example, GOx/HRP/BSA/CNT displayed 6 times the activity of a mixture of GOx/HRP/BSA, under otherwise identical conditions. Similarly, GOx/HRP/BSA/ZrP had 3.5 times the activity of the corresponding mixture of GOx/HRP/BSA, in the absence of the nanoplates. These robust nano-dispersions worked extraordinarily well as active bio-catalysts. These two kinds of fabricated biocatalyst dispersions are also highly stable.

Modeling and Designing Particle-Regulated Amyloid-like Assembly of Synthetic Polypeptides in Aqueous Solution.

In cells, actin and tubulin polymerization is regulated by nucleation factors, which promote the nucleation and subsequent growth of protein filaments in a controlled manner. Mimicking this natural mechanism to control the supramolecular polymerization of macromolecular monomers by artificially created nucleation factors remains a largely unmet challenge. Biological nucleation factors act as molecular scaffolds to boost the local concentrations of protein monomers and facilitate the required conformational changes to accelerate the nucleation and subsequent polymerization. An accelerated assembly of synthetic poly(l-glutamic acid) into amyloid fibrils catalyzed by cationic silica nanoparticle clusters (NPCs) as artificial nucleation factors is demonstrated here and modeled as supramolecular polymerization with a surface-induced heterogeneous nucleation pathway. Kinetic studies of fibril growth coupled with mechanistic analysis demonstrate that the artificial nucleators predictably accelerate the supramolecular polymerization process by orders of magnitude (e.g., shortening the assembly time by more than 10 times) when compared to the uncatalyzed reaction, under otherwise identical conditions. Amyloid-like fibrillation was supported by a variety of standard characterization methods. Nucleation followed a Michaelis-Menten-like scheme for the cationic silica NPCs, while the corresponding anionic or neutral nanoparticles had no effect on fibrillation. This approach shows the effectiveness of charge-charge interactions and surface functionalities in facilitating the conformational change of macromolecular monomers and controlling the rates of nucleation for fibril growth. Molecular design approaches like these inspire the development of novel materials via biomimetic supramolecular polymerizations.

One-step preparation of bioactive enzyme/inorganic materials.

Simultaneous exfoliation of crystalline α-zirconium phosphate (α-ZrP) nanosheets and enzyme binding, induced by shearing, without the addition of any toxic additives is reported here for the first time. These materials were thoroughly characterized and used for applications. The bulk α-ZrP material (20 mg mL-1) was exfoliated with low concentrations of a protein such as bovine serum albumin (BSA, 3 mg mL-1) in a shear reactor at 10k rpm for <80 minutes. Exfoliation was monitored by powder X-ray diffraction with samples displaying a gradual but complete loss of the 7.6 Å (002) peak, which is characteristic of bulk α-ZrP. The fully exfoliated sample loaded with the protein was characterized by transmission and scanning electron microscopy in addition to other biophysical methods. Lysozyme, glucose oxidase, met-hemoglobin, and ovalbumin also induced exfoliation and directly produced enzyme/ZrP biocatalysts. Thus, exfoliation, biophilization and enzyme binding are accomplished in a single step. Several factors contributed to the exfoliation kinetics, and the rate increased with α-ZrP and BSA concentrations and decreased with pH. However, the exfoliation efficiency inversely depended on the isoelectric point of the protein with ovalbumin (pI = 4.5) being the best and lysozyme (pI = 11.1) being the worst. A strong correlation between the protein size and exfoliation efficiency was noted, and the latter suggests the role of hydrodynamic factors in the process. Exfoliation was also achieved by simple stirring using a magnetic stirrer, under low volumes, and model enzymes, indicating 60-90% retention of bound enzymatic activities. The addition of BSA to enzymes as the diluent and stabilizing agent also prevents enzymes from the denaturing effect caused by stirring. This new method requires no pre-treatment of α-ZrP with toxic exfoliating agents such as tetrabutyl ammonium hydroxide and provides bioactive enzyme/inorganic materials in a single step. These protein-loaded biocompatible nanosheets may be useful for biocatalysis and biomedical applications.

Kinetic Study on Enzymatic Hydrolysis of Cellulose in an Open, Inhibition-Free System.

Due to the complexity of cellulases and the requirement of enzyme adsorption on cellulose prior to reactions, it is difficult to evaluate their reaction with a general mechanistic scheme. Nevertheless, it is of great interest to come up with an approximate analytic description of a valid model for the purpose of developing an intuitive understanding of these complex enzyme systems. Herein, we used the surface plasmonic resonance method to monitor the action of a cellobiohydrolase by itself, as well as its mixture with a synergetic endoglucanase, on the surface of a regenerated model cellulose film, under continuous flow conditions. We found a phenomenological approach by taking advantage of the long steady state of cellulose hydrolysis in the open, inhibition-free system. This provided a direct and reliable way to analyze the adsorption and reaction processes with a minimum number of fitting parameters. We investigated a generalized Langmuir-Michaelis-Menten model to describe a full set of kinetic results across a range of enzyme concentrations, compositions, and temperatures. The overall form of the equations describing the pseudo-steady-state kinetics of the flow-system shares some interesting similarities with the Michaelis-Menten equation. The use of familiar Michaelis-Menten parameters in the analysis provides a unifying framework to study cellulase kinetics. The strategy may provide a shortcut for approaching a quantitative while intuitive understanding of enzymatic degradation of cellulose from top to bottom. The open system approach and the kinetic analysis should be applicable to a variety of cellulases and reaction systems to accelerate the progress in the field.

Micelles Embedded in Multiphasic Protein Hydrogel Enable Efficient and Air-Tolerant Triplet Fusion Upconversion with Heavy-Atom and Spin-Orbit Charge-Transfer Sensitizers.

The applications of triplet-triplet annihilation-based photon upconversion (TTA-UC) in solar devices have been limited by the challenges in designing a TTA-UC system that is efficient under aerobic conditions. Efficient TTA-UC under aerobic conditions is typically accomplished by using soft matter or solid-state media, which succeed at protecting the triplet excited states of upconverters (sensitizer and annihilator) from quenching by molecular oxygen but fail at preserving their mobility, thus limiting the TTA-UC efficiency (ΦUC). We showcase a protein/lipid hydrogel that succeeded in doing both of the above due to its unique multiphasic design, with a high ΦUC of 19.0 ± 0.7% using a palladium octaethylporphyrin sensitizer. This hydrogel was made via an industrially compatible method using low-cost and eco-friendly materials: bovine serum albumin (BSA), sodium dodecyl sulfate (SDS), and water. A dense BSA network provided oxygen protection while the encapsulation of upconverters within a micellar SDS environment preserved upconverter mobility, ensuring near-unity triplet energy transfer efficiency. In addition to heavy atom-containing sensitizers, several completely organic, spin-orbit charge-transfer intersystem crossing (SOCT-ISC) Bodipy-based sensitizers were also studied; one of which achieved a ΦUC of 3.5 ± 0.2%, the only reported SOCT-ISC-sensitized ΦUC in soft matter to date. These high efficiencies showed that our multiphasic design was an excellent platform for air-tolerant TTA-UC and that it can be easily adapted to a variety of upconverters.

Exfoliated and water dispersible biocarbon nanotubes for enzymology applications.

In this chapter, we report a simple and facile method to armor enzymes with carbon nanotubes (CNTs) which are exfoliated, and debundled using bovine serum albumin (BSA). The fabricated CNT/BSA dispersions are biofriendly, biocompatible, defect-free, and highly stable solutions. BSA gives maximum exfoliation efficiency, exceeding the 4mg/mL of CNT concentration compared to any previous reports. Further, the produced bioCNT dispersions were characterized by UV-visible, Raman, circular dichroism spectroscopy, and scanning electron microscopy (SEM). Exfoliation and debundling of the bioCNT dispersions is possible due to the π-π interaction, hydrogen bonding, hydrophobic interaction, and electrostatic attractive forces driving the adsorption of BSA on CNTs surface. Protein adsorption then makes a highly stable suspension in water that can be stored for a prolonged period. CNT dispersions are stable over a wide range of pH from 3 to 10 and at 4°C or 25°C for more than 2 months. Here, we also report the facile, inexpensive and green-chemistry method to fabricate a buckypaper (CNT paper), composed of the high packing density, self-assembled and randomly oriented bioCNTs, and these assemblies could be used in many emerging applications like air and water purification, nanocomposites, energy storage, and biosensing. Moreover, the CNT dispersions stabilized by BSA were successfully used in enzyme binding and kinetic studies and bound enzyme retained substantial catalytic activity. The current approach may facilitate bulk production of water dispersed CNTs in both academic and industrial laboratories. This is done by a simple method of stirring, which provides new opportunities for a wider range of CNT applications.

Cerium Oxide Nanoparticles Improve Outcome after In Vitro and In Vivo Mild Traumatic Brain Injury.

Mild traumatic brain injury results in aberrant free radical generation, which is associated with oxidative stress, secondary injury signaling cascades, mitochondrial dysfunction, and poor functional outcome. Pharmacological targeting of free radicals with antioxidants has been examined as an approach to treatment, but has met with limited success in clinical trials. Conventional antioxidants that are currently available scavenge a single free radical before they are destroyed in the process. Here, we report for the first time that a novel regenerative cerium oxide nanoparticle antioxidant reduces neuronal death and calcium dysregulation after in vitro trauma. Further, using an in vivo model of mild lateral fluid percussion brain injury in the rat, we report that cerium oxide nanoparticles also preserve endogenous antioxidant systems, decrease macromolecular free radical damage, and improve cognitive function. Taken together, our results demonstrate that cerium oxide nanoparticles are a novel nanopharmaceutical with potential for mitigating neuropathological effects of mild traumatic brain injury and modifying the course of recovery.

Engineering functional inorganic nanobiomaterials: controlling interactions between 2D-nanosheets and enzymes.

A better understanding of the enzyme-nanosheet interface is imperative for the design of functional, robust inorganic nanobiomaterials and biodevices, now more than ever, for use in a broad spectrum of applications. This feature article discusses recent advances in controlling the enzyme-nanosheet interface with regards to α-zirconium(iv) phosphate (α-ZrP), graphene oxide (GO), graphene, and MoS2 nanosheets. Specific focus will be placed on understanding the mechanisms with which these materials interact with enzymes and elaborate on particular ways to engineer and control these interactions. Our main discoveries include: (1) upon adsorption to the nanosheet surface, a decrease in the entropy of the enzyme's denatured state enhances stability; (2) proteins are used to create biophilic landing pads for increased enzyme stability on many different types of nanosheets; (3) proteins and enzymes are used as exfoliants by shear force to produce biofunctionalized nanosheet suspensions; and (4) bionfunctionalized nanosheets exhibit no acute toxicity. Recognizing proper methods to engineer the interface between enzymes and 2D-nanosheets, therefore, is an important step towards making green, sustainable, and environmentally conscious inorganic bionanomaterials for sensing, catalysis and drug delivery applications, as well as towards the successful manipulation of enzymes for advanced applications.

Stirred Not Shaken: Facile Production of High-Quality, High-Concentration Graphene Aqueous Suspensions Assisted by a Protein.

A simple method to produce record concentrations (up to 10 mg mL-1) of high-quality aqueous graphene suspensions by using an ordinary benchtop magnetic stirrer is reported. The shear rates employed here are almost 10 times less than those in previous reports, and graphene is efficiently separated from unexfoliated graphite during the synthesis. Systematic optimization of synthesis parameters, such as pH, protein concentration, temperature, stirrer speed, and volume of solution, afforded efficient conversion (100%) of graphite to graphene-aqueous suspensions. The synthesis is readily scaled-up with a continuous flow reactor where the graphene is produced and separated 24/7, with little or no human intervention. Raman spectroscopy confirmed little to no sp3 or oxidative defects, and that the graphene nanosheets consisted of three to five layers. The graphene suspensions were coated on aluminum and tested for thermal conductivity applications. The thermal conductivity of our graphene sample was calculated to be 684 W m-1 K-1, a value greater than that of a commercial sample. The activation energy measured for shear exfoliation by stirring was found to be over 45 billion times smaller than the corresponding thermal activation energy, affording physical insight into the process. We hypothesize that stirring selectively populates translational states that are necessary for exfoliation and thus requires far less energy than conventional exfoliation methods, where the energy is uniformly distributed among all available modes. Therefore, an efficient, convenient, and inexpensive method for graphene production in limited-resource settings is reported here.

Interlocking Enzymes in Graphene-Coated Cellulose Paper for Increased Enzymatic Efficiency.

A simple method for interlocking glucose oxidase and horseradish peroxidase in a network of cellulose fibers coated with bovine serum albumin (BSA)-exfoliated graphene (biographene) is reported here. The resulting paper reactor is inexpensive and stable. Biographene is expected to function as an electron shuttle, making the reaction between the enzyme and the substrate more efficient, and this hypothesis is examined here. The BSA used to separate the sheets of graphene provides extra carboxylic acid groups and primary amines to help interlock the enzymes and the graphene in between the fibers. The decrease in entropy associated with interlocking the enzymes on a solid support is likely responsible for the increase in enzymatic stability/activity observed. Each cellulose disk contained 5.2mg of enzyme per gram of paper and 93% of the enzyme is retained after washing for 0.5-2h. This simple methodology provides a low cost, effective approach for achieving high enzymatic activity and good loadings on a benign, versatile support.

A Simple Flow Reactor for Continuous Synthesis of Biographene for Enzymology Studies.

The unique properties of graphene make it an intriguing platform for the attachment and enhancement of biological molecules, but it has yet to achieve its full potential in terms of biological applications. Single-layer graphene is expensive, making alternatives to this material highly desired for applications that require high-quality graphene in large quantities. In this context, we report a simple, environmentally friendly, nonlabor-intensive method for the synthesis of colloidal graphene suspensions of 3-5 layers, stabilized by bovine serum albumin, in water. The method involves a flow reactor designed to continually yield high-quality graphene colloids, synthesized, purified, and optimized all in one setup. The flow reactor is able to produce colloidal graphene sheets on a multigram scale, and these colloids were characterized by Raman spectroscopy, electron microscopy, and zeta potential studies. The average size of the sheets is 0.16µm2, each consisting of 3-5 layers of graphene with little or no sp3 defects. These graphene colloids stabilized by the protein were successfully used in protein kinetic studies as well as in surface plasmon resonance protein binding studies. The ease of synthesis of these high-quality graphene colloidal suspensions in water provides an exciting opportunity for biographene to be used on an industrial scale for electronic, thermal, and enzymology applications.