Novel TFG mutation causes autosomal-dominant spastic paraplegia and defects in autophagy.

BACKGROUND: Mutations in the tropomyosin receptor kinase fused (TFG) gene are associated with various neurological disorders, including autosomal recessive hereditary spastic paraplegia (HSP), autosomal dominant hereditary motor and sensory neuropathy with proximal dominant involvement (HMSN-P) and autosomal dominant type of Charcot-Marie-Tooth disease type 2. METHODS: Whole genome sequencing and whole-exome sequencing were used, followed by Sanger sequencing for validation. Haplotype analysis was performed to confirm the inheritance mode of the novel TFG mutation in a large Chinese family with HSP. Additionally, another family diagnosed with HMSN-P and carrying the reported TFG mutation was studied. Clinical data and muscle pathology comparisons were drawn between patients with HSP and patients with HMSN-P. Furthermore, functional studies using skin fibroblasts derived from patients with HSP and patients with HMSN-P were conducted to investigate the pathomechanisms of TFG mutations. RESULTS: A novel heterozygous TFG variant (NM_006070.6: c.125G>A (p.R42Q)) was identified and caused pure HSP. We further confirmed that the well-documented recessively inherited spastic paraplegia, caused by homozygous TFG mutations, exists in a dominantly inherited form. Although the clinical features and muscle pathology between patients with HSP and patients with HMSN-P were distinct, skin fibroblasts derived from both patient groups exhibited reduced levels of autophagy-related proteins and the presence of TFG-positive puncta. CONCLUSIONS: Our findings suggest that autophagy impairment may serve as a common pathomechanism among different clinical phenotypes caused by TFG mutations. Consequently, targeting autophagy may facilitate the development of a uniform treatment for TFG-related neurological disorders.

Fluorescent probe based on GO/g-C3N4-PEG@Cu NPs/MIP for the detection of dopamine in banana.

Graphene oxide (GO) and copper nanoparticles (Cu NPs) were incorporated to modulate and enhance the fluorescence properties of pegylated graphite phase carbon nitride (g-C3N4-PEG). Combined with the specific recognition capability of a molecular imprinted polymer (MIP), a highly sensitive and selective fluorescent molecular imprinted probe for dopamine detection was developed. The fluorescent g-C3N4-PEG was synthesized from melamine and modified with GO and Cu NPs to obtain GO/g-C3N4-PEG@Cu NPs. Subsequently, MIP was prepared on the surface of GO/g-C3N4-PEG@Cu NPs using dopamine as the template molecule. Upon elution of the template molecule, a dopamine-specific GO/g-C3N4-PEG@Cu NPs/MIP fluorescence probe was obtained. The fluorescence intensity of the probe was quenched through the adsorption of different concentrations of dopamine by the MIP, thus establishing a novel method for the detection of dopamine. The linear range of dopamine detection was from 5 × 10-11 to 6 × 10-8 mol L-1, with a detection limit of 2.32 × 10-11 mol L-1. The sensor was utilised for the detection of dopamine in bananas, achieving a spiked recovery rate between 90.3% and 101.3%. These results demonstrate that the fluorescence molecular imprinted sensor developed in this study offers a highly sensitive approach for dopamine detection in bananas.

Silicone elastomer gel impregnated with 20(S)-protopanaxadiol-loaded nanostructured lipid carriers for ordered diabetic ulcer recovery.

Inefficient diabetic ulcer healing and scar formation remain a challenge worldwide, owing to a series of disordered and dynamic biological events that occur during the process of healing. A functional wound dressing that is capable of promoting ordered diabetic wound recovery is eagerly anticipated. In this study, we designed a silicone elastomer with embedded 20(S)-protopanaxadiol-loaded nanostructured lipid carriers (PPD-NS) to achieve ordered recovery in scarless diabetic ulcer healing. The nanostructured lipid carriers were prepared through an emulsion evaporation-solidification method and then incorporated into a network of silicone elastomer to form a unique nanostructured lipid carrier-enriched gel formulation. Interestingly, the PPD-NS showed excellent in vitro anti-inflammatory and proangiogenic activity. Moreover, in diabetic mice with full-thickness skin excision wound, treatment with PPD-NS significantly promoted in vivo scarless wound healing through suppressing inflammatory infiltration in the inflammatory phase, promoting angiogenesis during the proliferation phase, and regulating collagen deposition in the remodeling phase. Hence, this study demonstrates that the developed PPD-NS could facilitate ordered diabetic wound recovery via multifunctional improvement during different wound-healing phases. This novel approach could be promising for scarless diabetic wound healing.

Uniaxially aligned electrospun all-cellulose nanocomposite nanofibers reinforced with cellulose nanocrystals: scaffold for tissue engineering.

Uniaxially aligned cellulose nanofibers with well oriented cellulose nanocrystals (CNCs) embedded were fabricated via electrospinning using a rotating drum as the collector. Scanning electron microscope (SEM) images indicated that most cellulose nanofibers were uniaxially aligned. The incorporation of CNCs into the spinning dope resulted in more uniform morphology of the electrospun cellulose/CNCs nanocomposite nanofibers (ECCNN). Polarized light microscope (PLM) and transmission electron microscope (TEM) showed that CNCs dispersed well in ECCNN nonwovens and achieved considerable orientation along the long axis direction. This unique hierarchical microstructure of ECCNN nonwovens gave rise to remarkable enhancement of their physical properties. By incorporating 20% loading (in weight) of CNCs, the tensile strength and elastic modulus of ECCNN along the fiber alignment direction were increased by 101.7 and 171.6%, respectively. Their thermal stability was significantly improved as well. In addition, the ECCNN nonwovens were assessed as potential scaffold materials for tissue engineering. It was elucidated from MTT tests that the ECCNN were essentially nontoxic to human cells. Cell culture experiments demonstrated that cells could proliferate rapidly not only on the surface but also deep inside the ECCNN. More importantly, the aligned nanofibers of ECCNN exhibited a strong effect on directing cellular organization. This feature made the scaffold particularly useful for various artificial tissues or organs, such as blood vessel, tendon, nerve, and so on, in which cell orientation was crucial for their performance.

Ultrasound microbubble-mediated miR-503-5p downregulation suppressed in vitro CRC progression via promoting SALL1 expression.

BACKGROUND: This study compared the effect of ultrasound microbubble-mediated miR-503-5p downregulation with that of pure liposome-mediated miR-503-5p downregulation on colorectal cancer (CRC) progression and explored the downstream mechanism. METHODS: Bioinformatics tools were utilized to predict miR-503-5p-targeted genes and CRC progression-associated genes. MiR-503-5p and sal-like 1 (SALL1) expressions in CRC cells and tissues were analyzed by qRT-PCR and/or bioinformatics tools; their correlations with overall survival and clinicopathological features of CRC patients were presented, and their interaction was validated by dual-luciferase reporter assay. CRC cells received ultrasound microbubble-mediated miR-503-5p downregulation and/or liposome-mediated miR-503-5p downregulation or SALL1 silencing. Cell phenotype changes were evaluated by flow cytometry, as well as MTT, Wound healing, Transwell and tube formation assays. E-cadherin, N-cadherin, Vimentin, B-cell lymphoma (Bcl)- 2, Cleaved caspase-3, and SALL1 expressions in cells were analyzed by Western blot. RESULTS: Upregulated miR-503-5p in CRC tissues and cells was detected, associated with poorer cell differentiation, easier lymph node metastasis and higher TNM stages, and related to poorer prognoses of CRC patients. Ultrasound microbubble-mediated miR-503-5p downregulation relative to pure liposome-mediated miR-503-5p downregulation better decreased viability, inhibited migration, invasion and tube formation, enhanced apoptosis, upregulated SALL1, E-cadherin and Cleaved caspase-3, and downregulated miR-503-5p, N-cadherin, Vimentin and Bcl-2 in CRC cells. SALL1 was targeted by miR-503-5p, low-expressed in CRC tissues and cells and positively related to CRC patients' survival. Silencing SALL1 exerted the opposite effects, which reversed the effects of ultrasound microbubble-mediated miR-503-5p downregulation and vice versa. CONCLUSION: Ultrasound microbubble-mediated miR-503-5p downregulation suppressed in vitro CRC progression via promoting SALL1 expression.

Highly Effective Stroke Therapy Enabled by Genetically Engineered Viral Nanofibers.

Stroke results in the formation of a cavity in the infarcted brain tissue. Angiogenesis and neurogenesis are poor in the cavity, preventing brain-tissue regeneration for stroke therapy. To regenerate brain tissue in the cavity, filamentous phages, the human-safe nanofiber-like bacteria-specific viruses, are genetically engineered to display many copies of RGD peptide on the sidewalls. The viral nanofibers, electrostatically coated on biocompatible injectable silk protein microparticles, not only promote adhesion, proliferation, and infiltration of neural stem cells (NSCs), but also induce NSCs to differentiate preferentially into neurons in basal medium within 3 d. After the NSC-loaded microparticles are injected into the stroke cavity of rat models, the phage nanofibers on the microparticles stimulate angiogenesis and neurogenesis in the stroke sites within two weeks for brain regeneration, leading to functional recovery of limb motor control of rats within 12 weeks. The viral nanofibers also brought about the desired outcomes for stroke therapy, such as reducing inflammatory response, decreasing thickness of astrocytes scars, and increasing neuroblasts response in the subventricular zone. As virtually any functional peptide can be displayed on the phage by genetic means, the phage nanofibers hold promise as a unique and effective injectable biomaterial for stroke therapy.

One-Step Construct Responsive Lignin/Polysaccharide/Fe Nano Loading System Driven by Digestive Enzymes of Lepidopteran for Precise Delivery of Pesticides.

A strategy that relies on the differences in feeding behavior between pests and natural enemies to deliver insecticides precisely was proposed. After proving that the digestive enzymes in Lepidopteran pests can act as triggers for lignin-based controlled-release carriers, a novel multiple-enzyme-responsive lignin/polysaccharide/Fe nanocarrier was constructed by combining the electrostatic self-assembly and chelation and loaded with lambda-cyhalothrin (LC) to form a nanocapsule suspension loading system. The nanocapsules were LC@sodium lignosulfonate/chitosan/Fe (LC@SL/CS/Fe) and LC@sodium lignosulfonate/alkyl polyglycoside quaternary ammonium salt/Fe (LC@SL/APQAS/Fe). LC@SL/APQAS/Fe was more stable than LC@SL/CS/Fe because it adsorbs more Fe3+, and the half-lives of LC in LC@SL/APQAS/Fe under UV irradiation were prolonged at 4.02- and 6.03-folds than those of LC@SL/CS/Fe and LC emulsifiable concentrate (LC EC), respectively. Both LC@SL/APQAS/Fe and LC@SL/CS/Fe have responsive release functions to laccase and cellulase, and the release rate of the former was slower. The insecticidal activity of LC@SL/APQAS/Fe against Agrotis ipsilonis was similar to those of LC@SL/CS/Fe and LC EC, while the toxicity of LC@SL/APQAS/Fe to the natural enemy was 2-3 times less than those of LC@SL/CS/Fe and LC EC. Meanwhile, the organic solvent component in the nanocapsule suspension was 94% less than that in the EC preparation. Therefore, the nano loading system based on SL/APQAS/Fe is a promising nanoplatform with the advantages of high efficiency, low toxicity, and environmental friendliness.

Boosting enzymatic degradation of cellulose using a fungal expansin: Structural insight into the pretreatment mechanism.

The recalcitrance of cellulosic biomass greatly hinders its enzymatic degradation. Expansins induce cell wall loosening and promote efficient cellulose utilization; however, the molecular mechanism underlying their action is not well understood. In this study, TlEXLX1, a fungal expansin from Talaromyces leycettanus JCM12802, was characterized in terms of phylogeny, synergy, structure, and mechanism of action. TlEXLX1 displayed varying degrees of synergism with commercial cellulase in the pretreatment of corn straw and filter paper. TlEXLX1 binds to cellulose via domain 2, mediated by CH-π interactions with residues Tyr291, Trp292, and Tyr327. Residues Asp237, Glu238, and Asp248 in domain 1 form hydrogen bonds with glucose units and break the inherent hydrogen bonding within the cellulose matrix. This study identified the expansin amino acid residues crucial for cellulose binding, and elucidated the structure and function of expansins in cell wall networks; this has potential applications in biomass utilization.

Air-plasma treatment promotes bone-like nano-hydroxylapatite formation on protein films for enhanced in vivo osteogenesis.

Introducing hydroxylapatite (HAp) into biomolecular materials is a promising approach to improve their bone regenerative capability. Thus a facile method needs to be developed to achieve this goal. Here we show that a simple air-plasma treatment of silk fibroin (SF) films for 5 min induced the formation of bone-like plate-shaped nano-HAp (nHAp) on their surface and the resultant material efficiently enhanced in vivo osteogenesis. The air-plasma-treated SF films (termed A-SF) presented surface nano-pillars and enhanced hydrophilicity compared to the pristine SF films (termed SF), making the A-SF and SF films induce the formation of plate-shaped/more-crystalline and needle-like/less-crystalline nHAp, respectively. The mineralized A-SF and SF films (termed A-SF-nHAp and SF-nHAp, respectively) and their non-mineralized counterparts were seeded with rat mesenchymal stem cells and subcutaneously implanted into the rat models. The A-SF-nHAp and A-SF films exhibited more efficient bone formation than the SF-nHAp and SF films in 4 weeks due to their unique nanotopography, with the A-SF-nHAp films being more efficient than the A-SF films. This work shows that a combination of the air-plasma treatment and the subsequent nHAp mineralization most efficiently promotes bone formation. Our plasma-based method is an attractive approach to enhance the bone regenerative capacity of protein-based biomaterials.

Facile surface-engineered polymeric absorbents for simultaneous adsorption and degradation of organic wastes.

Polymeric absorbents were surface-engineered with photocatalysts for simultaneous adsorption and degradation of organic wastes in water using a facile Pickering emulsion polymerization method. This facile strategy not only overcome the low light energy utilization of traditional photocatalysts-polymer nanocomposites due to the core-shell structure, but also could convenient control the microstructure of the photocatalysts owing to the separated preparation procedure when compared to the direct growth method. Firstly, binary bismuth oxyhalide composed as BiOI0.5Cl0.5 was chosen to engineered polymeric absorbents due to its higher photodegradation efficiency especially. After the emulsification and polymerization process, BiOI0.5Cl0.5 acting as the stabilizer would be fixed on the surface of the functional polymeric absorbents to form PA@BiOI0.5Cl0.5. Batch experiments were lunched using phenol as the test substance, the results shown that PA@BiOI0.5Cl0.5 could remove phenol completely within a short time and could be reused without any treatments with a simultaneous adsorption and degradation process. Furthermore, polymeric absorbents were engineered with commercial TiO2 to prove the generality of this strategy.

Water repellent Ag/Ag2O@bamboo cellulose fiber membrane as bioinspired cargo carriers.

Water striders can walk on water. To mimic this function, a porous membrane consisted of bamboo cellulose fiber was hybridized with Ag/Ag2O nanoparticles through a facile in situ method to produce water repellent and well-ventilated materials. Herein, we report the sole surface roughness created by Ag/Ag2O nanoparticles could render the membrane a water contact angle (CA) of 140±3.0°. When floating on water, the hybrid membrane was able to support a heavy load more than 10 times the weight of the membrane itself. Additionally, this membrane demonstrated capabilities for oil sampling under water or oil/water separation and strong antibacterial activity against Escherichia coli. Thus we foresee that this novel hybrid membrane can be potentially utilized as drag-reducing, gas permeable and antibiotic substrates for constructing miniature aquatic devices.

Reinforcement of all-cellulose nanocomposite films using native cellulose nanofibrils.

All-cellulose nanocomposite films were prepared using native cellulose nanofibrils (CNFs) as fillers and lithium chloride/N,N-dimethylacetamide (LiCl/DMAc) dissolved regenerated cellulose as the matrix. The CNFs, with diameters in the range of 15-40 nm were obtained by combined physical methods of ultrasonic treatment and high shear homogenization. The morphology, structure, and properties of the nanocomposite films were characterized by scanning electron microscope (SEM), X-ray diffraction (XRD), optical transmittance, thermal gravimetric analysis (TGA), and mechanical testing. The nanocomposite films exhibited good optical transparency, thermal stability, and remarkably enhanced mechanical properties compared to the regenerated cellulose matrix. By varying the CNFs content, the tensile strength of the nanocomposite films increased from 61.56 MPa to 99.92 MPa and the Young's modulus increased from 0.76 GPa to 4.16 GPa. This work provided a promising pathway for manufacturing high performance and environmental-friendly all-cellulose nanocomposites.

Allosteric analysis of glucocorticoid receptor-DNA interface induced by cyclic Py-Im polyamide: a molecular dynamics simulation study.

BACKGROUND: It has been extensively developed in recent years that cell-permeable small molecules, such as polyamide, can be programmed to disrupt transcription factor-DNA interfaces and can silence aberrant gene expression. For example, cyclic pyrrole-imidazole polyamide that competes with glucocorticoid receptor (GR) for binding to glucocorticoid response elements could be expected to affect the DNA dependent binding by interfering with the protein-DNA interface. However, how such small molecules affect the transcription factor-DNA interfaces and gene regulatory pathways through DNA structure distortion is not fully understood so far. METHODOLOGY/PRINCIPAL FINDINGS: In the present work, we have constructed some models, especially the ternary model of polyamides+DNA+GR DNA-binding domain (GRDBD) dimer, and carried out molecular dynamics simulations and free energy calculations for them to address how polyamide molecules disrupt the GRDBD and DNA interface when polyamide and protein bind at the same sites on opposite grooves of DNA. CONCLUSIONS/SIGNIFICANCE: We found that the cyclic polyamide binding in minor groove of DNA can induce a large structural perturbation of DNA, i.e. a >4 Šwidening of the DNA minor groove and a compression of the major groove by more than 4 Šas compared with the DNA molecule in the GRDBD dimer+DNA complex. Further investigations for the ternary system of polyamides+DNA+GRDBD dimer and the binary system of allosteric DNA+GRDBD dimer revealed that the compression of DNA major groove surface causes GRDBD to move away from the DNA major groove with the initial average distance of ∼4 Što the final average distance of ∼10 Šduring 40 ns simulation course. Therefore, this study straightforward explores how small molecule targeting specific sites in the DNA minor groove disrupts the transcription factor-DNA interface in DNA major groove, and consequently modulates gene expression.

A thermophilic cellulase complex from Phialophora sp. G5 showing high capacity in cellulose hydrolysis.

A cellulase-producing mesophilic fungal strain, named G5, was isolated from the acidic wastewater and mud of a tin mine and identified as Phialophora sp. based on the internal transcribed spacer sequence. The volumetric activities and specific activities of cellulase induced by different carbon sources (Avicel, corn cob, wheat bran and corn stover) were compared. The cellulase complex of Phialophora sp. G5 exhibited the optimal activities at 60-65 °C and pH 4.0-5.0, and had good long-term thermostability at 50 °C. Compared with the commercial cellulase (Accellerase 1500, Genencor), the enzyme under study showed 60% and 80% of the capacity to hydrolyze pure cellulose and natural cellulose, respectively. This is the first study to report that a cellulytic enzymes complex from Phialophora genus, and the superior properties of this enzyme complex make strain G5 a potential microbial source to produce cellulase for industrial applications, and the production ability could be improved by mutagenesis.