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Nanobodies derived from camelids and sharks offer unique advantages in therapeutic applications due to their ability to bind to epitopes that were previously inaccessible. Traditional methods of nanobody development face challenges such as ethical concerns and antigen toxicity. Our study presents a synthetic, phagedisplayed nanobody library using trinucleotide-directed mutagenesis technology, which allows precise amino acid composition in complementarity-determining regions (CDRs), with a focus on CDR3 diversity. This approach avoids common problems such as frameshift mutations and stop codon insertions associated with other synthetic antibody library construction methods. By analyzing FDA-approved nanobodies and Protein Data Bank sequences, we designed sub-libraries with different CDR3 lengths and introduced amino acid substitutions to improve solubility. The validation of our library through the successful isolation of nanobodies against targets such as PD-1, ATXN1 and STAT3 demonstrates a versatile and ethical platform for the development of high specificity and affinity nanobodies and represents a significant advance in biotechnology.
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This paper proposes a retinal prosthesis edge detection (RPED) algorithm that can achieve high visual acuity and low power. Retinal prostheses have been used to stimulate retinal tissue by injecting charge via an electrode array, thereby artificially restoring the vision of visually impaired patients. The retinal prosthetic chip, which generates biphasic current pulses, should be located in the foveal area measuring 5 mm × 5 mm. When a high-density stimulation pixel array is realized in a limited area, the distance between the stimulation pixels narrows, resulting in current dispersion and high-power dissipation related to heat generation. Various edge detection methods have been proposed over the past decade to reduce these deleterious effects and achieve high-resolution pixels. However, conventional methods have the disadvantages of high-power consumption and long data processing times because many pixels are activated to detect edges. In this study, we propose a novel RPED algorithm that has a higher visual acuity and less power consumption despite using fewer active pixels than existing techniques. To verify the performance of the devised RPED algorithm, the peak signal-to-noise ratio and structural similarity index map, which evaluates the quantitative numerical value of the image are employed and compared with the Sobel, Canny, and past edge detection algorithms in MATLAB. Finally, we demonstrate the effectiveness of the proposed RPED algorithm using a 1600-pixel retinal stimulation chip fabricated using a 0.35 µm complementary metal-oxide-semiconductor process.
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Algoritmos , Acuidade Visual , Próteses Visuais , Humanos , Acuidade Visual/fisiologia , Retina/fisiologia , Retina/diagnóstico por imagem , Razão Sinal-RuídoRESUMO
USP47 is widely involved in tumor development, metastasis, and other processes while performing a more regulatory role in inflammatory responses, myocardial infarction, and neuronal development. In this study, we investigate the functional and biochemical properties of USP47, whereby depleting USP47 inhibited cancer cell growth in a p53-dependent manner-a phenomenon that enhances during the simultaneous knockdown of USP7. Full-length USP47 shows higher deubiquitinase activity than the catalytic domain. The crystal structures of the catalytic domain, in its free and ubiquitin-bound states, reveal that the misaligned catalytic triads, ultimately, become aligned upon ubiquitin-binding, similar to USP7, thereby becoming ready for catalysis. Yet, the composition and lengths of BL1, BL2, and BL3 of USP47 differ from those for USP7, and they contribute to the observed selectivity. Our study provides molecular details of USP47 regulation, substrate recognition, and the hotspots for drug discovery by targeting USP47.
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Transformação Celular Neoplásica , Ubiquitina , Humanos , Peptidase 7 Específica de Ubiquitina/genética , Catálise , Domínio CatalíticoRESUMO
In this study, a pulse frequency modulation (PFM)-based stimulator is proposed for use in biomedical implantable devices. Conventionally, functional electrical stimulation (FES) techniques have been used to reinforce damaged nerves, such as retina tissue and brain tissue, by injecting a certain amount of charge into tissues. Although several design methods are present for implementing FES devices, an FES stimulator for retinal implants is difficult to realize because of the chip area, which needs to be inserted in a fovea, sized 5 mm x 5 mm, and power limitations to prevent the heat generation that causes tissue damage. In this work, we propose a novel stimulation structure to reduce the compliance voltage during stimulation, which can result in high-speed and low-voltage operation. A new stimulator that is composed of a modified high-speed PFM, a 4-bit counter, a serializer, a digital controller, and a current driver is designed and verified using a DB HiTek standard 0.18 µm process. This proposed stimulator can generate a charge up to 130 nC, consumes an average power of 375 µW during a stimulation period, and occupies a total area of 700 µm × 68 µm.
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Terapia por Estimulação Elétrica , Próteses Visuais , Eletrodos Implantados , Retina , Fóvea Central , Estimulação Elétrica , Desenho de EquipamentoRESUMO
(-)-Epigallocatechin gallate (EGCG) is one of the autophagy stimulators that have been reported to protect vascular endothelial cells from oxidative stress-induced damage. In this study, we attempted potentiation of the autophagy-stimulating activity of EGCG in human aortic epithelial cells (HAECs) by using the EGCG-phenylalanine conjugate, E10. Autophagy-stimulating activity of E10 was evaluated by LC3-II measurement in the absence and presence of the lysosomal blocker chloroquine, CTYO-ID staining, and reporter assay using tandem fluorescence-tagged LC3. These experiments revealed significantly enhanced autophagic flux stimulation in HAECs by E10 compared with EGCG. Further elaboration of E10 showed that activation of AMPK through phosphorylation as the major mechanism of its autophagy stimulation. Like other autophagy stimulators, E10 protected HAECs from lipotoxicity as well as accompanying endothelial senescence. Finally, stimulation of autophagy by E10 was shown to protect HAECs from oxidative stress-induced apoptosis. These findings collectively suggest potential clinical implications of E10 for various cardiovascular complications through stimulation of autophagy.
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Catequina , Células Endoteliais , Humanos , Fenilalanina/farmacologia , Chá , Catequina/farmacologia , Catecóis , AutofagiaRESUMO
In this study, we propose a low-area multi-channel controlled dielectric breakdown (CDB) system that simultaneously produces several nanopore sensors. Conventionally, solid-state nanopores are prepared by etching or drilling openings in a silicon nitride (SiNx) substrate, which is expensive and requires a long processing time. To address these challenges, a CDB technique was introduced and used to fabricate nanopore channels in SiNx membranes. However, the nanopore sensors produced by the CDB result in a severe pore-to-pore diameter variation as a result of different fabrication conditions and processing times. Accordingly, it is indispensable to simultaneously fabricate nanopore sensors in the same environment to reduce the deleterious effects of pore-to-pore variation. In this study, we propose a four-channel CDB system that comprises an amplifier that boosts the command voltage, a 1-to-4 multiplexer, a level shifter, a low-noise transimpedance amplifier and a data acquisition device. To prove our design concept, we used the CDB system to fabricate four nanopore sensors with diameters of <10 nm, and its in vitro performance was verified using λ-DNA samples.
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Nanoporos , Nanotecnologia , DNA , Nanotecnologia/métodos , Sistemas Automatizados de Assistência Junto ao LeitoRESUMO
Several imidazolium-based ionic liquids significantly enhance the activity of MoO2 for electrochemical reduction of CO2. The overpotential of CO2 reduction is as low as 40 mV. The ionic liquids act as both electrolytes and co-catalysts, which not only leads to lower overpotentials, but also alters the product selectivity.
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The clinical efficacy of conventional chemotherapeutic agent, methotrexate (MTX), can be limited by its very short plasma half-life, the drug resistance, and the high dosage required for cancer cell suppression. In this study, a new drug delivery system is proposed to overcome such limitations. To realize such a system, MTX was intercalated into layered double hydroxides (LDHs), inorganic drug delivery vehicle, through a co-precipitation route to produce a MTX-LDH nanohybrid with an average particle size of approximately 130 nm. Biodistribution studies in mice bearing orthotopic human breast tumors revealed that the tumor-to-liver ratio of MTX in the MTX-LDH-treated-group was 6-fold higher than that of MTX-treated-one after drug treatment for 2 hr. Moreover, MTX-LDH exhibited superior targeting effect resulting in high antitumor efficacy inducing a 74.3% reduction in tumor volume compared to MTX alone, and as a consequence, significant survival benefits. Annexin-V and propidium iodine dual staining and TUNEL analysis showed that MTX-LDH induced a greater degree of apoptosis than free MTX. Taken together, our data demonstrate that a new MTX-LDH nanohybrid exhibits a superior efficacy profile and improved distribution compared to MTX alone and has the potential to enhance therapeutic efficacy via inhibition of tumor proliferation and induction of apoptosis.
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Silicatos de Alumínio/química , Antimetabólitos Antineoplásicos/farmacologia , Neoplasias da Mama/tratamento farmacológico , Hidróxido de Magnésio/química , Glândulas Mamárias Animais/efeitos dos fármacos , Metotrexato/farmacologia , Animais , Anexina A5 , Antimetabólitos Antineoplásicos/química , Apoptose/efeitos dos fármacos , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacos , Portadores de Fármacos , Composição de Medicamentos , Feminino , Meia-Vida , Xenoenxertos , Humanos , Fígado/efeitos dos fármacos , Glândulas Mamárias Animais/patologia , Metotrexato/química , Camundongos , Transplante de Neoplasias , Tamanho da Partícula , Propídio , Carga Tumoral/efeitos dos fármacosRESUMO
MoO2 microparticles act as an active catalyst for the electrochemical reduction of CO2 in organic solvents such as acetonitrile and dimethylformamide. The catalytic activity and product selectivity depend on temperature and water content of the solvent.
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Poor aqueous solubility and the unpleasant taste of aripiprazole (APZ) have been recurring problems, owing to its low bioavailability and low patient tolerance, respectively. Herein, we prepared a nanohybrid system that was based on a bentonite clay material, montmorillonite (MMT), which could both mask the taste and enhance the solubility of APZ (i.e., APZ-MMT). To further improve the efficacy of this taste masking and drug solubility, APZ-MMT was also coated with a cationic polymer, polyvinylacetal diethylamino acetate (AEA). In vitro dissolution tests at neutral pH showed that the amount of drug that was released from the AEA-coated APZ-MMT was greatly suppressed (<1%) for the first 3 min, thus suggesting that AEA-coated APZ-MMT has strong potential for the taste masking of APZ. Notably, in simulated gastric juice at pH 1.2, the total percentage of APZ that was released within the first 2 h increased up to 95% for AEA-coated APZ-MMT. Furthermore, this in vitro release profile was also similar to that of Abilify®, a commercially available medication. In vivo experiments by using Sprague-Dawley rats were also performed to compare the pharmacokinetics of AEA-coated APZ-MMT and Abilify®. AEA-coated APZ-MMT exhibited about 20% higher systemic exposure of APZ and its metabolite, dehydro-APZ, compared with Abilify®. Therefore, a new MMT-based nanovehicle, which is coated with a cationic polymer, can act as a promising delivery system for both taste masking and for enhancing the bioavailability of APZ.
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Bentonita/farmacologia , Piperazinas/farmacologia , Quinolonas/farmacologia , Animais , Aripiprazol , Bentonita/química , Bentonita/farmacocinética , Disponibilidade Biológica , Humanos , Concentração de Íons de Hidrogênio , Masculino , Nanoestruturas , Piperazinas/química , Piperazinas/farmacocinética , Polímeros/química , Quinolonas/química , Quinolonas/farmacocinética , Ratos , Ratos Sprague-Dawley , Solubilidade , PaladarRESUMO
Reduction of CO(2) by electrochemical and photoelectrochemical methods to produce carbon-rich fuels is a heavily pursued research theme. Most of the current efforts are focused on the development of transition-metal-based catalysts. In this tutorial review, we present an overview of the development of organic molecules as mediators and catalysts for CO(2) reduction. Four classes of organic molecules are discussed: tetraalkylammonium salts, aromatic esters and nitriles, ionic liquids, and pyridinium derivatives. It is shown that reactions mediated or catalyzed by these organic molecules can be competitive compared to their metal-catalyzed counterparts, both in terms of product selectivity and energy efficiency.
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A nanohybrid was prepared with an inorganic clay material, montmorillonite (MMT), for taste masking of sildenafil (SDN). To further improve the taste-masking efficiency and enhance the drug-release rate, we coated the nanohybrid of SDN-MMT with a basic polymer, polyvinylacetal diethylaminoacetate (AEA). Powder X-ray diffraction and Fourier transform infrared experiments showed that SDN was successfully intercalated into the interlayer space of MMT. The AEA-coated SDN-MMT nanohybrid showed drug release was much suppressed at neutral pH (release rate, 4.70 ± 0.53%), suggesting a potential for drug taste masking at the buccal cavity. We also performed in vitro drug release experiments in a simulated gastric fluid (pH = 1.2) and compared the drug-release profiles of AEA-coated SDN-MMT and Viagra(®), an approved dosage form of SDN. As a result, about 90% of SDN was released from the AEA-coated SDN-MMT during the first 2 hours while almost 100% of drug was released from Viagra(®). However, an in vivo experiment showed that the AEA-coated SDN-MMT exhibited higher drug exposure than Viagra(®). For the AEA-coated SDN-MMT, the area under the plasma concentration- time curve from 0 hours to infinity (AUC(0-∞)) and maximum concentration (C(max)) were 78.8 ± 2.32 µg · hour/mL and 12.4 ± 0.673 µg/mL, respectively, both of which were larger than those obtained with Viagra(®) (AUC(0-∞) = 69.2 ± 3.19 µg · hour/mL; C(max) = 10.5 ± 0.641 µg/mL). Therefore, we concluded that the MMT-based nanohybrid is a promising delivery system for taste masking of SDN with possibly improved drug exposure.
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Nanoconjugados/administração & dosagem , Piperazinas/administração & dosagem , Sulfonas/administração & dosagem , Paladar , Administração Oral , Animais , Bentonita/química , Preparações de Ação Retardada/administração & dosagem , Preparações de Ação Retardada/química , Cães , Portadores de Fármacos/química , Estabilidade de Medicamentos , Masculino , Nanoconjugados/química , Nanomedicina , Piperazinas/sangue , Piperazinas/farmacocinética , Polivinil/química , Difração de Pó , Purinas/administração & dosagem , Purinas/sangue , Purinas/farmacocinética , Citrato de Sildenafila , Espectroscopia de Infravermelho com Transformada de Fourier , Sulfonas/sangue , Sulfonas/farmacocinéticaRESUMO
Delivery of poorly soluble drugs has been problematic due to its low absorption profile and bioavailability. In this work, ursodeoxycholic acid (UDCA), a poorly-soluble drug, was intercalated into inorganic nanovehicle, layered double hydroxides (LDHs), with a molecular level to enhance its solubility in biological fluid. The UDCA-loaded nanovehicle (i.e., UDCA-LDHs) was also coated with an anionic polymer, Eudragit(®) S100, to increase the dissolution rate of UDCA. According to the powder X-ray diffraction (PXRD) patterns of UDCA-LDHs, the gallery height of LDHs was expanded from 3.6Å to 28.3Å, indicating that the UDCA molecules were successfully intercalated into the interlayer space of LDHs. Fourier transform infrared (FT-IR) spectra also revealed that the UDCA molecules were well stabilized in the LDHs through electrostatic interaction. The in vitro dissolution test in a simulated biological fluid (pH=6.8) showed that the total dissolved fraction of UDCA for the first 2h was about 60.2% for the Eudragit(®) S100 coated UDCA-LDHs, which was a dramatic increase as compared with 19.0% dissolution from intact UDCA. It is, therefore, concluded that LDHs nanovehicle coated with an anionic polymer is a promising delivery system for improving aqueous solubility of poorly soluble drugs.