Enhanced trans-cleavage activity using CRISPR-Cas12a variant designed to reduce steric inhibition by cis-cleavage products.

The CRISPR-Cas12a system has emerged as a promising tool for molecular diagnostics due to its indiscriminate trans-ssDNase activity. However, the sensitivity of Cas12a-based diagnostics remains insufficient for clinical use without a pre-amplification step such as loop-mediated isothermal amplification, and therefore the trans-cleavage activity of Cas12a needs to be enhanced. Here, we present a novel strategy to enhance the trans-cleavage activity of Cas12a by reducing the steric hindrance from cis-cleavage products.

Emerging strategies to fabricate polymeric nanocarriers for enhanced drug delivery across blood-brain barrier: An overview.

Blood-brain barrier (BBB) serves as an essential interface between central nervous system (CNS) and its periphery, allowing selective permeation of ions, gaseous molecules, and other nutrients to maintain metabolic functions of brain. Concurrently, it restricts passage of unsolicited materials from bloodstream to CNS which could otherwise lead to neurotoxicity. Nevertheless, in the treatment of neurodegenerative diseases such as Parkinson's, Alzheimer's, diffuse intrinsic pontine glioma, and other brain cancers, drugs must reach CNS.

pH-responsive polyelectrolyte complexation on upconversion nanoparticles: a multifunctional nanocarrier for protection, delivery, and 3D-imaging of therapeutic protein.

The delicate tertiary structure of proteins, their susceptibility to heat- and enzyme-induced irreversible denaturation, and their tendency to get accumulated at the cell membrane during uptake are daunting challenges in proteinaceous therapeutic delivery. Herein, a polyelectrolyte complex having encapsulated therapeutic protein has been designed on the surface of upconverting luminescent nanoparticles (NaYF:20%Yb,2%Er). This nanosized complex system has been found to overcome the challenges of protein aggregation at the cell membrane.

Effect of Immersive Virtual Reality-Based Bilateral Arm Training in Patients with Chronic Stroke.

Virtual reality (VR)-based therapies are widely used in stroke rehabilitation. Although various studies have used VR techniques for bilateral upper limb training, most have been only semi-immersive and have only been performed in an artificial environment. This study developed VR content and protocols based on activities of daily living to provide immersive VR-based bilateral arm training (VRBAT) for upper limb rehabilitation in stroke patients.

Uptake of Polyelectrolyte Functionalized Upconversion Nanoparticles by Tau-Aggregated Neuron Cells.

Tauopathy is the aggregation phenomenon of tau proteins and associated with neurodegenerative diseases. It metastasizes via the transfer of tau aggregates to adjacent neuron cells; however, the mechanism has not yet been fully understood. Moreover, if the materials used for designing drug delivery system to treat such neurodegenerative diseases do not undergo biodegradation or exocytosis but remains in cells or tissues, they raise concerns about their possible negative impacts.

Structural basis for the inhibition of PDK2 by novel ATP- and lipoyl-binding site targeting compounds.

Pyruvate dehydrogenase kinase (PDK) controls the activity of pyruvate decarboxylase complex (PDC) by phosphorylating key serine residues on the E1 subunit, which leads to a decreased oxidative phosphorylation in mitochondria. Inhibition of PDK activity by natural/synthetic compounds has been shown to reverse the Warburg effect, a characteristic metabolism in cancer cells. PDK-PDC axis also has been associated with diabetes and heart disease.

Anomalous Dynamics of in Vivo Cargo Delivery by Motor Protein Multiplexes.

Vesicle transport conducted by motor protein multiplexes (MPMs), which is ubiquitous among eukaryotes, shows anomalous and stochastic dynamics qualitatively different from the dynamics of thermal motion and artificial active matter; the relationship between in vivo vesicle-delivery dynamics and the underlying physicochemical processes is not yet quantitatively understood. Addressing this issue, we perform accurate tracking of individual vesicles, containing upconverting nanoparticles, transported by kinesin-dynein-multiplexes along axonal microtubules. The mean-square-displacement of vesicles along the microtubule exhibits unusual dynamic phase transitions that are seemingly inconsistent with the scaling behavior of the mean-first-passage time over the travel length.

Two-Dimensional and Three-Dimensional Single Particle Tracking of Upconverting Nanoparticles in Living Cells.

Lanthanide-doped upconversion nanoparticles (UCNPs) are inorganic nanomaterials in which the lanthanide cations embedded in the host matrix can convert incident near-infrared light to visible or ultraviolet light. These particles are often used for long-term and real-time imaging because they are extremely stable even when subjected to continuous irradiation for a long time. It is now possible to image their movement at the single particle level with a scale of a few nanometers and track their trajectories as a function of time with a scale of a few microseconds.

Osteoconductivity of Binary Titanium Alloys with Different Micro/Nanoporous Surfaces.

The surface characteristics and osteoconductivity were evaluated for the micro/nanoporous surfaces of titanium (Ti) alloys prepared by micro-arc oxidation (MAO) and hydrothermal treatment (HT) of binary Ti-5 wt% A alloys (A = Au, Mn, Nb, and Pd). Surface properties were analyzed using X-ray diffractometry, scanning electron microscopy, and energy dispersive X-ray spectroscopy. The osteoconductivity was evaluated by measuring the total protein, ALPase activity, and osteocalcin production.

Cellular uptake efficiency of nanoparticles investigated by three-dimensional imaging.

Understanding the interaction of nanoparticles with living cells on the basis of cellular uptake efficiency is a fundamental requisite in biomedical research. Cellular internalization of nanoparticles takes place by mechanisms like ATP hydrolysis-driven endocytosis that deliver nanoparticles to the cytoplasm, organelles and nuclei. Despite its importance in nanomedicine, this uptake procedure is not understood in-depth because of the complexity of the biochemical mechanisms and the lack of available experimental methods for quantitative analysis.

Osteoblast cell adhesion and viability on nanostructured surfaces of porous titanium oxide layer.

Surface characteristics and osteoblast cell functions were investigated for the nano-structured oxide layer on commercially pure titanium (CP-Ti) fabricated using microarc oxidation (MAO) and hydrothermal treatment (HT) methods. Ti-MAO-135HT, Ti-MAO-150HT, and Ti-MAO-175HT groups were fabricated by hydrothermally treating the MAO-treated specimens (Ti-MAO) in phosphorus-containing alkaline solution at temperatures of 135, 150, or 175 °C, respectively. After hydrothermal treatment, a nanosheet-shaped morphology, nano-needles and nanorods were observed on the porous surface of the Ti-MAO-135HT, Ti-MAO-150HT and Ti-MAO-175HT groups, respectively.

Cytotoxicity of alloying elements and experimental titanium alloys by WST-1 and agar overlay tests.

Objective: This study was performed to evaluate the biocompatibility of nine types of pure metals using 36 experimental prosthetic titanium-based alloys containing 5, 10, 15, and 20wt% of each substituted metal.

Methods: The cell viabilities for pure metals on Ti alloys that contain these elements were compared with that of commercially pure (CP) Ti using the WST-1 test and agar overlay test.

Results: The ranking of pure metal cytotoxicity from most potent to least potent was: Co>Cu>In>Ag>Cr>Sn>Au>Pd>Pt>CP Ti.

Cytotoxicity of soft denture lining materials depending on their component types.

Purpose: To evaluate the difference in cytotoxicity of soft denture lining materials depending on their component types.

Materials And Methods: Ten commercially available soft denture lining materials (SDLM) consisting of five silicone-based materials and five acrylic-based materials were evaluated. For the MTT test, cured SDLM samples were extracted in a culture medium for 24 hours, and L-929 cells were incubated in the extracted medium for 24 hours.

Cytocompatibility of pure metals and experimental binary titanium alloys for implant materials.

Objective: This study was performed to evaluate the biocompatibility of nine types of pure metal ingots (Ag, Al, Cr, Cu, Mn, Mo, Nb, V, Zr) and 36 experimental titanium (Ti) alloys containing 5, 10, 15, and 20 wt% of each alloying element.

Methods: The cell viabilities for each test group were compared with that of CP-Ti using the WST-1 test and agar overlay test.

Results: The ranking of pure metal cytotoxicity from most potent to least potent was as follows: Cu>Al>Ag>V>Mn>Cr>Zr>Nb>Mo>CP-Ti.

Chemical composition and antimicrobial activity of the essential oil of Cryptomeria japonica.

The composition and the antibacterial activity of the essential oil obtained from Cryptomeria japonica D. Don on oral bacteria were studied. The chemical composition of the essential oil was analysed by GC and GC-MS.

Genes of periodontopathogens expressed during human disease.

Background: Since many bacterial genes are environmentally regulated, the screening for virulence-associated factors using classical genetic and molecular biology approaches can be biased under laboratory growth conditions of a given pathogen, because the required conditions for expression of many virulence factors may not occur during in vitro growth. Thus, technologies have been developed during the past several years to identify genes that are expressed during disease using animal models of human disease. However, animal models are not always truly representative of human disease, and with many pathogens, there is no appropriate animal model.

Chemical composition and antimicrobial activity of the essential oil of Artemisia lavandulaefolia.

The chemical components of the essential oil obtained from Artemisia lavandulaefolia DC. were analyzed by GC-MS. Ninety-nine compounds accounting for 94.

Chemical composition and antimicrobial activity of the essential oils of Artemisia scoparia and A. capillaris.

The chemical composition of the essential oils obtained from Artemisia scoparia Waldst. et Kitamura and Artemisia capillaris Thunb. was analyzed by GC/MS.