Proteomic Analysis of Marinesco–Sjogren Syndrome Fibroblasts Indicates Pro-Survival Metabolic Adaptation to SIL1 Loss

Marinesco–Sjogren syndrome (MSS) is a rare multisystem pediatric disorder, caused by loss-of-function mutations in the gene encoding the endoplasmic reticulum cochaperone SIL1. SIL1 acts as a nucleotide exchange factor for BiP, which plays a central role in secretory protein folding. SIL1 mutant cells have reduced BiP-assisted protein folding, cannot fulfil their protein needs, and experience chronic activation of the unfolded protein response (UPR). Maladaptive UPR may explain the cerebellar and skeletal muscle degeneration responsible for the ataxia and muscle weakness typical of MSS. However, the cause of other more variable, clinical manifestations, such as mild to severe mental retardation, hypogonadism, short stature, and skeletal deformities, is less clear. To gain insights into the pathogenic mechanisms and/or adaptive responses to SIL1 loss, we carried out cell biological and proteomic investigations in skin fibroblasts derived from a young patient carrying the SIL1 R111X mutation. Despite fibroblasts not being overtly affected in MSS, we found morphological and biochemical changes indicative of UPR activation and altered cell metabolism. All the cell machineries involved in RNA splicing and translation were strongly downregulated, while protein degradation via lysosome-based structures was boosted, consistent with an attempt of the cell to reduce the workload of the endoplasmic reticulum and dispose of misfolded proteins. Cell metabolism was extensively affected as we observed a reduction in lipid synthesis, an increase in beta oxidation, and an enhancement of the tricarboxylic acid cycle, with upregulation of eight of its enzymes. Finally, the catabolic pathways of various amino acids, including valine, leucine, isoleucine, tryptophan, lysine, aspartate, and phenylalanine, were enhanced, while the biosynthetic pathways of arginine, serine, glycine, and cysteine were reduced. These results indicate that, in addition to UPR activation and increased protein degradation, MSS fibroblasts have profound metabolic alterations, which may help them cope with the absence of SIL1.     

Nondestructive evaluation of cork enclosures using terahertz/millimeter wave spectroscopy and imaging

Natural cork enclosures, due to their cell structure, composition, and low moisture are fairly transparent to terahertz (THz) and millimeter waves enabling nondestructive evaluation of the cork's surface and interior. It is shown that the attenuation coefficient of the defect-free cork can be modeled with a Mie scattering model in the weakly scattering limit. Contrast in the THz images is a result of enhanced scattering of THz radiation by defects or voids as well as variations in the cork cell structure. The presence of voids, defects, and changes in grain structure can be determined with roughly 100-300 microm resolution.     

Controlled Release of Epigenetically-Enhanced Extracellular Vesicles from a GelMA/Nanoclay Composite Hydrogel to Promote Bone Repair

Extracellular vesicles (EVs) have garnered growing attention as promising acellular tools for bone repair. Although EVs’ potential for bone regeneration has been shown, issues associated with their therapeutic potency and short half-life in vivo hinders their clinical utility. Epigenetic reprogramming with the histone deacetylase inhibitor Trichostatin A (TSA) has been reported to promote the osteoinductive potency of osteoblast-derived EVs. Gelatin methacryloyl (GelMA) hydrogels functionalised with the synthetic nanoclay laponite (LAP) have been shown to effectively bind, stabilise, and improve the retention of bioactive factors. This study investigated the potential of utilising a GelMA-LAP hydrogel to improve local retention and control delivery of epigenetically enhanced osteoblast-derived EVs as a novel bone repair strategy. LAP was found to elicit a dose-dependent increase in GelMA compressive modulus and shear-thinning properties. Incorporation of the nanoclay was also found to enhance shape fidelity when 3D printed compared to LAP-free gels. Interestingly, GelMA hydrogels containing LAP displayed increased mineralisation capacity (1.41-fold) (p ≤ 0.01) over 14 days. EV release kinetics from these nanocomposite systems were also strongly influenced by LAP concentration with significantly more vesicles being released from GelMA constructs as detected by a CD63 ELISA (p ≤ 0.001). EVs derived from TSA-treated osteoblasts (TSA-EVs) enhanced proliferation (1.09-fold), migration (1.83-fold), histone acetylation (1.32-fold) and mineralisation (1.87-fold) of human bone marrow stromal cells (hBMSCs) when released from the GelMA-LAP hydrogel compared to the untreated EV gels (p ≤ 0.01). Importantly, the TSA-EV functionalised GelMA-LAP hydrogel significantly promoted encapsulated hBMSCs extracellular matrix collagen production (≥1.3-fold) and mineralisation (≥1.78-fold) in a dose-dependent manner compared to untreated EV constructs (p ≤ 0.001). Taken together, these findings demonstrate the potential of combining epigenetically enhanced osteoblast-derived EVs with a nanocomposite photocurable hydrogel to promote the therapeutic efficacy of acellular vesicle approaches for bone regeneration.     

Non-Destructive Measurement of Water Diffusion in Natural Cork Enclosures Using Terahertz Spectroscopy and Imaging

Terahertz (THz) imaging enables non-destructive evaluation of many materials’ internal structures which could not be probed by visual analysis alone. In this paper, we apply THz imaging to non-destructively probe and image the diffusion of water through natural cork samples. The temporal and spatial distribution of water is analyzed as it diffuses throughout the cork structure. An average diffusion coefficient comparable to previous work in the field is extracted from this analysis. Data is also presented to show that the diffusion coefficient is not uniform throughout the cork but changes dramatically due to the local structure and composition of the cork.     

Evaluation of Cu, Mn, and Se in Vegetables Using Ultrasonic Extraction and GFAAS Quantification

This paper proposes a method for the determination of copper, manganese, and selenium in vegetables of conventional and organic farming in the state of S?o Paulo/Brazil using ultrasonic extraction of analytes and subsequent quantification by graphite furnace atomic absorption spectrometry (GFAAS). Using 0.10 mol L⁻¹ of hydrochloric acid as the extracting solution, the following optimal extraction conditions were established: sample mass of 100 mg, sample grain size of <60 μm, sonication time of five cycles of 40 s, and sonication power of 136 W. The analytes were determined by GFAAS, applying drying temperatures of 90 °C to 250 °C, pyrolysis temperature of 1,300 °C, atomization temperature of 2,300 °C, and cleaning temperature of 2,800 °C. Palladium nitrate was co-injected with the samples as a chemical modifier and tungsten as a permanent modifier. The accuracy and precision of the proposed extraction method were evaluated based on a certified standard, RM 8433 Corn Bran (NIST RM 8433). The results obtained by the ultrasonic extraction method were equivalent to those obtained by the method of acid mineralization of samples in a microwave oven. However, the proposed methodology reduces the analytical time considerably, favoring the analytical speed. Moreover, the quantity of residues generated and discharged into the environment is also considerably lower.     

Experimental comparison of performance degradation from terahertz and infrared wireless links in fog

We describe a lab setup for analyzing impairments of terahertz (THz) and infrared (IR) free space links caused by local refraction index changes in the signal's propagation paths that could be induced by turbulence, particles, humidity, etc. A THz signal comprising a 2.5 Gb/s data load modulated on a carrier at 625 GHz, is launched through a weather emulating chamber, detected, and its performance analyzed. An IR beam at 1.5 um wavelength carrying the same data load is superposed with the THz beam, propagating through the same weather conditions and also performance analyzed. We modulate the IR channel with a usual non-return-to-zero (NRZ) format but use duobinary coding for driving our THz source, which enables signaling at high data rate and higher output power. As both beams pass through the same channel perturbations and as their degradations are recorded simultaneously we can simultaneously compare the weather impact on both. We investigate scintillation and fog attenuation effects for the THz and IR signals by measuring bit error rates (BER), signal power, and phase front distortions.     

Polarized light reflection from strained sinusoidal surfaces

We propose optical polarization imaging as a minimally invasive technique for measuring the mechanical properties of plastics and soft tissues through their change in reflectance properties with applied strain or force. We suggest that changes in surface roughness are responsible for the linear reflectivity changes with applied stretch or strain. Several aspects of this model are tested, including the dependence on the angle of incidence, the change in scattering and absorption coefficients with strain, and the lateral spatial resolution. The application of the technique to multilayer structures such as skin and competing optical effects such as laser speckle are discussed.     

Formulation development and human in vitro-in vivo correlation for a novel, monolithic controlled-release matrix system of high load and highly water-soluble drug niacin

Novel, controlled-release formulations for high drug load, highly water soluble compound niacin based on polyethylene oxide (PEO) and hydroxypropylmethyl cellulose (HPMC) matrices were developed and investigated. The effect of sodium bicarbonate as a modulator of swelling, erosion, and drug release and its impact on changes in the kinetics of axial swelling and gel strength were evaluated by textural analysis during dissolution study. The drug release rate from PEO-based matrices was faster and correlated with lower gel strength, greater water uptake, and greater matrix erosion. Slower release rate and greater release duration correlated significantly with greater matrix swelling with negligible matrix erosion for the HPMC-based matrix system. Inclusion of sodium bicarbonate in the polymeric matrix salted out the macromolecules and increased gel strength and gel viscosity, especially in the vicinity of the swelling fronts. An in vivo study in human subjects after administration of the formulations and a commercial product exhibited similar plasma concentrations. For the formulation of interest, the mean drug fraction absorbed by the body was calculated by the Wagner-Nelson technique, and a level A “in vitro-in vivo correlation” was observed between the percent released in vitro and percent absorbed in vivo. The developed formulations appear to be robust and easy to manufacture with maximum flexibility with respect to drug dose, polymeric carriers, duration, and kinetics of drug release.     

Lentinus (Panus) tigrinus augmentation of a historically contaminated soil: matrix decontamination and structure and function of the resident bacterial community

The ability of Lentinus tigrinus to grow and to degrade persistent aromatic hydrocarbons in aged contaminated soil was assessed in this study. L. tigrinus extensively colonized the soil; its degradation activity after 60 d incubation at 28°C, however, was mostly limited to dichloroaniline isomers, polychlorinated benzenes and diphenyl ether while the fungus was unable to deplete 9,10-anthracenedione and 7-H-benz[DE]anthracene-7-one which were the major soil contaminants. Although clean-up levels were limited, both density of cultivable heterotrophic bacteria and richness of the resident bacterial community in L. tigrinus microcosms (LtM) increased over time to a significantly larger extent than the respective amended incubation controls (1.9×10(9) CFU g(-1) vs. 1.0×10(9) CFU g(-1) and 37 vs. 16, respectively). Naphthalene- and catechol 2,3-dioxygenase gene copy numbers, however, decreased over time at a higher rate in LtM than in incubation controls likely due to a higher stimulation on heterotrophs than xenobiotics-degrading community members.     

Interactions amongHeliothis virescens larvae,cotton condensed tannin and the CryIA(c) δ-endotoxin ofBacillus thuringiensis

The potential interactions among a plant-produced allelochemical, a phytophagous insect, and an endotoxin produced byBacillus thuringiensis were investigated using purified cotton condensed tannins, the CryIA(c)-endotoxin fromB. thuringiensis subsp.kurstaki strain HD-73, and larvae ofHeliothis virescens. Purified condensed tannin from cotton fed to neonateH. virescens reduced feeding and mortality caused by insecticidal crystals ofB. thuringiensis. In fifth instars, tannin reduced relative growth rate (RGR), relative consumption rate (RCR), but antagonized the effects of the crystal-endotoxin. Tannin did not deter feeding of fifth instars in choice tests with cellulose-ester disks. Masking tannin from interacting with the dietary ingredients of artificial diets and crystal protein by encapsulation in alginate gel suggested that tannin adversely affected feeding after ingestion.These results suggest that insect control tactics that employ-endotoxins in microbial insecticides and transgenic cotton plants may not be compatible when used in conjunction with plants containing high tannin concentrations.