Sugar Glasses Research Articles

ISOLASI DAN INTENSITAS SERANGAN PENYAKIT BULAI (Peronosclerospora philippinensis Shaw) PADA TANAMAN JAGUNG (Zea mays L.) DI KABUPATEN MAROS

This study aims to determine the isolation and intensity of downy mildew (Peronosclerospora philippinensis) on maize (Zea mays L.) in Maros Regency during the 2022 planting period. This research was carried out on agricultural land in Pucak Village, Tompobulu District, Maros Regency and at the Pest Laboratory. and Diseases of the Indonesian Muslim University and the Hasanuddin University Disease Laboratory. This research was carried out for 3 months, starting from June to August 2022. The materials and tools used were leaves that were attacked by downy mildew, 2% sugar water, methylene blue, aquades, microscopes, scissors, glass objects, masking tape, tissue towels, plastic clips, camera and stationery. This study uses a survey method by direct observation in the field. Downy mildew leaves are taken and then taken to the laboratory to identify the fungus that causes downy mildew. Based on microscopic observations, Peronosclerospora philippinensis species were found with ovoid shape and thin walls. The results of the observation that the intensity of downy mildew in the field in the first month was 20% with the moderate attack intensity category, then in the second month it increased to 70.2% with the severe category and in the third month which was the highest attack intensity with an average of 96.7%. with the puso category which means that of the 30 plants observed, 29 were attacked by downy mildew. The attack intensity which reaches 96.7% can no longer be used as a seed source.

Hybrid Lipid Nanocapsules: A Robust Platform for mRNA Delivery.

The success of the mRNA vaccine against COVID-19 has garnered significant interest in the development of mRNA therapeutics against other diseases, but there remains a strong need for a stable and versatile delivery platform for these therapeutics. In this study, we report on a family of robust hybrid lipid nanocapsules (hLNCs) for the delivery of mRNA. The hLNCs are composed of kolliphore HS15, labrafac lipophile WL1349, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), and a conjugate of oleic acid (OA) and polyethylenimines of varying size (PEI─0.8, 1.8, and 25 kDa). They are prepared by a solvent-free, temperature-phase inversion method, yielding an average size of ∼40 nm and a particle distribution index (PDI) < 0.2. We demonstrate that the PDI remains <0.2 over a wide pH range and in a wide range of medium. We further show that the PDI and the functionality of mRNA condensed on the particles are robust to drying in a sugar glass and subsequent rehydration. Finally, we demonstrate that mRNA-loaded hLNCs yield reasonable transfection in vitro and in vivo settings.

Mechanical, Morphological, and Fire Behaviors of Sugar Palm/Glass Fiber Reinforced Epoxy Hybrid Composites

This research aims to investigate using sugar palm fiber (SPF) and glass fiber (GF) in an epoxy matrix to develop composite materials with improved mechanical, morphological, and flammability properties. The mechanical and flammability properties are examined per ASTM standards, while the morphological study examines the fractured surfaces of the samples. Using the hand lay-up technique, the hybrid composite comprises 15% SPF, 15% GF, and 70% epoxy resin. Three treatments are applied to the SPF: untreated, alkaline treated, and benzoyl chloride treated, which enables research into the effect of fiber treatment on mechanical properties and flammability. The morphological investigation reveals that both treated SPF/GF/EP composites exhibit lower tensile strength than the untreated SPF/GF/EP composite due to inadequate mechanical interlocking at the fiber-matrix interface. However, the alkaline-treated SPF/GF/EP composite demonstrates a 24.8% improvement in flexural strength, a 1.52% increase in impact strength, and a 9.76% enhancement in flammability. Similarly, the benzoyl chloride-treated SPF/GF/EP composite improves flexural strength, impact strength, and flammability by 24.6%, 0.51%, and 5.66%, respectively. These results highlight the potential of fiber treatment to improve composite materials’ mechanical and flammability properties.

Stabilization of a flavoprotein for solid-state photo-CIDNP MAS NMR at room temperature by embedding in a glassy sugar matrix

Hyperpolarization via the solid-state photochemically induced dynamic nuclear polarization (photo-CIDNP) effect can be detected in frozen solutions of electron transfer proteins generating a radical-pair upon illumination. The effect has been observed in various natural photosynthetic reaction centers and in light-oxygen-voltage (LOV) sensing domains incorporating a flavin mononucleotide (FMN) as chromophore. In LOV domains, where a highly conserved cysteine is mutated to a flavin to interrupt its natural photochemistry, a radical-pair is generated by electron transfer from a nearby tryptophan to the photoexcited triplet state of FMN. During the photocycle, both the LOV domain and the chromophore are photochemically degraded, e.g., by the formation of singlet oxygen. This limits the time for collection of hyperpolarized nuclear magnetic resonance (NMR) data. We show that embedding of the protein into a trehalose sugar glass matrix stabilizes the protein for 13C solid-state photo-CIDNP NMR experiments which can be conducted at room temperature in a powder sample. Additionally, this preparation allows for incorporation of high amounts of protein further boosting the intensity of the detected signals from FMN and tryptophan at natural abundance. Signal assignment is aided by quantum chemical calculations of absolute shieldings. The underlying mechanism for the surprising absorption-only signal pattern is not yet understood. Comparison to calculated isotropic hyperfine couplings imply that the enhancement is not due to the classical radical-pair mechanism (RPM). Analysis of the anisotropic hyperfine couplings associated with solid-state photo-CIDNP mechanisms also show no simple correlation, suggesting a more complex underlying mechanism.

Mechanism of protein stabilization in crowded solution and preservation in desiccated sugar glass.

The highly crowded cellular milieu includes countless different macromolecules and osmolytes aimed at regulating osmotic stress while protecting proteins and other macromolecules. Of special interest is the formation of cellular glassy matrices composed of sugars, which strongly correlate with the ability of some yeast, nematodes, and tardigrades to survive desiccation and extreme temperature changes. Remarkably, these carbohydrates stabilize proteins in crowded solutions and preserve them in the dry glassy state. Yet, although considerable efforts have been directed to discern how crowding stabilizes proteins, the molecular-level stabilization mechanism has remained elusive. We have followed the effect of various osmolytes on the native state stability of model miniproteins in crowded solutions using circular dichroism (CD) spectroscopy and our newly developed mean-field theory for crowding. We show that protein native state stability and folding thermodynamic signature can be explained using three distinct types of interactions: excluded volume, solvent-osmolyte non-ideal mixing, and osmolyte-protein soft-interactions. Furthermore, using our combination of synchrotron radiation CD spectroscopy together with sampling enhanced molecular dynamics (MD) simulations we have been able to shed light on the stability and new emerging nanostructures of proteins when they are embedded in sugar glass.

Biofabrication of Sodium Alginate Hydrogel Scaffolds for Heart Valve Tissue Engineering.

Every year, thousands of aortic valve replacements must take place due to valve diseases. Tissue-engineered heart valves represent promising valve substitutes with remodeling, regeneration, and growth capabilities. However, the accurate reproduction of the complex three-dimensional (3D) anatomy of the aortic valve remains a challenge for current biofabrication methods. We present a novel technique for rapid fabrication of native-like tricuspid aortic valve scaffolds made of an alginate-based hydrogel. Using this technique, a sodium alginate hydrogel formulation is injected into a mold produced using a custom-made sugar glass 3D printer. The mold is then dissolved using a custom-made dissolving module, revealing the aortic valve scaffold. To assess the reproducibility of the technique, three scaffolds were thoroughly compared. CT (computed tomography) scans showed that the scaffolds respect the complex native geometry with minimal variations. The scaffolds were then tested in a cardiac bioreactor specially designed to reproduce physiological flow and pressure (aortic and ventricular) conditions. The flow and pressure profiles were similar to the physiological ones for the three valve scaffolds, with small variabilities. These early results establish the functional repeatability of this new biofabrication method and suggest its application for rapid fabrication of ready-to-use cell-seeded sodium alginate scaffolds for heart valve tissue engineering.

Improving the 3D Printability of Sugar Glass to Engineer Sacrificial Vascular Templates.

A prominent obstacle in scaling up tissue engineering technologies for human applications is engineering an adequate supply of oxygen and nutrients throughout artificial tissues. Sugar glass has emerged as a promising 3D-printable, sacrificial material that can be used to embed perfusable networks within cell-laden matrices to improve mass transfer. To characterize and optimize a previously published sugar ink, we investigated the effects of sucrose, glucose, and dextran concentration on the glass transition temperature (Tg), printability, and stability of 3D-printed sugar glass constructs. We identified a sucrose ink formulation with a significantly higher Tg (40.0 ± 0.9°C) than the original formulation (sucrose-glucose blend, Tg = 26.2 ± 0.4°C), which demonstrated a pronounced improvement in printability, resistance to bending, and final print stability, all without changing dissolution kinetics and decomposition temperature. This formulation allowed printing of 10-cm-long horizontal cantilever filaments, which can enable the printing of complex vascular segments along the x-, y-, and z-axes without the need for supporting structures. Vascular templates with a single inlet and outlet branching into nine channels were 3D printed using the improved formulation and subsequently used to generate perfusable alginate constructs. The printed lattice showed high fidelity with respect to the input geometry, although with some channel deformation after alginate casting and gelation-likely due to alginate swelling. Compared with avascular controls, no significant acute cytotoxicity was noted when casting pancreatic beta cell-laden alginate constructs around improved ink filaments, whereas a significant decrease in cell viability was observed with the original ink. The improved formulation lends more flexibility to sugar glass 3D printing by facilitating the fabrication of larger, more complex, and more stable sacrificial networks. Rigorous characterization and optimization methods for improving sacrificial inks may facilitate the fabrication of functional cellular constructs for tissue engineering, cellular biology, and other biomedical applications.

Characterisation of factors contributing to the performance of nonwoven fibrous matrices as substrates for adenovirus vectored vaccine stabilisation

Adenovirus vectors offer a platform technology for vaccine development. The value of the platform has been proven during the COVID-19 pandemic. Although good stability at 2–8 °C is an advantage of the platform, non-cold-chain distribution would have substantial advantages, in particular in low-income countries. We have previously reported a novel, potentially less expensive thermostabilisation approach using a combination of simple sugars and glass micro-fibrous matrix, achieving excellent recovery of adenovirus-vectored vaccines after storage at temperatures as high as 45 °C. This matrix is, however, prone to fragmentation and so not suitable for clinical translation. Here, we report an investigation of alternative fibrous matrices which might be suitable for clinical use. A number of commercially-available matrices permitted good protein recovery, quality of sugar glass and moisture content of the dried product but did not achieve the thermostabilisation performance of the original glass fibre matrix. We therefore further investigated physical and chemical characteristics of the glass fibre matrix and its components, finding that the polyvinyl alcohol present in the glass fibre matrix assists vaccine stability. This finding enabled us to identify a potentially biocompatible matrix with encouraging performance. We discuss remaining challenges for transfer of the technology into clinical use, including reliability of process performance.

β-Hairpin Miniprotein Stabilization in Trehalose Glass Is Facilitated by an Emergent Compact Non-Native State.

From stem cell freeze-drying to organ storage, considerable recent efforts have been directed toward the development of new preservation technologies. A prominent protein stabilizing strategy involves vitrification in glassy matrices, most notably those formed of sugars such as the biologically relevant preservative trehalose. Here, we compare the folding thermodynamics of a model miniprotein in solution and in the glassy state of the sugars trehalose and glucose. Using synchrotron radiation circular dichroism (SRCD), we find that the same native structure persists in solution and glass. However, upon transition to the glass, a completely different, conformationally restricted unfolded state replaces the disordered denatured state found in solution, potentially inhibiting misfolding. Concomitantly, a large exothermic contribution is observed in glass, exposing the stabilizing effect of interactions with the sugar matrix on the native state. Our results shed light on the mechanism of protein stabilization in sugar glass and should aid in future preservation technologies.

Infliximab formulation strategy for a stable ileo-colonic targeted oral dosage form intended for the topical treatment of inflammatory bowel disease

Research shows that topically active infliximab therapy may be efficacious in the treatment of inflammatory bowel disease (IBD) with expected fewer side effects related to systemic exposure. Oral administration of infliximab with site-specific delivery could enable such therapy. In the present study, infliximab was incorporated in a sugar glass matrix (IFX-I) for additional stability and compounding flexibility after which IFX-I was compounded to ileo-colonic-targeted tablets containing 5 mg infliximab (ColoPulse-IFX). Potential critical steps in the production process that may decrease the formulation stability were identified and investigated. Furthermore, the long-term stability of IFX-I (6 months) and ColoPulse-IFX (12 months) stored either at room temperature (25 °C ± 2 °C/60% RH ± 5% RH) or refrigerated (5 °C ± 3 °C) was investigated according to ICH guidelines. Size-exclusion chromatography, fluorescence spectroscopy, and ELISA analyses were used to investigate the infliximab stability, content, tertiary protein structure, and potency at t0, t3, t6, t9, and t12 months. The coating performance of ColoPulse-IFX was investigated in a gastrointestinal simulation system at t0 and t12 months. All the analyses showed that IFX-I and ColoPulse-IFX were stable, potent, and that the coating performance was maintained during the entire storage period at both storage conditions. Thus, IFX-I is a stable dry-powder formulation and ColoPulse-IFX is a promising oral dosage form for the topical treatment of ileo-colonic IBD. This formulation strategy may serve as a new platform for the development of oral peptide or protein formulations that are targeted to the ileo-colonic region in IBD.

Stabilization of Dry Sucrose Glasses by Four LEA_4 Proteins from Arabidopsis thaliana.

Cells of many organisms and organs can withstand an (almost) total water loss (anhydrobiosis). Sugars play an essential role in desiccation tolerance due to their glass formation ability during dehydration. In addition, intrinsically disordered LEA proteins contribute to cellular survival under such conditions. One possible mechanism of LEA protein function is the stabilization of sugar glasses. However, little is known about the underlying mechanisms. Here we used FTIR spectroscopy to investigate sucrose (Suc) glass stability dried from water or from two buffer components in the presence of four recombinant LEA and globular reference proteins. Buffer ions influenced the strength of the Suc glass in the order Suc < Suc/Tris < Suc/NaP. LEA proteins strengthened the sugar H-bonded network and the molecular structure in the glassy state. The position of νOH peak and the wavenumber–temperature coefficient (WTCg) provided similar information about the H-bonded network. Protein aggregation of LEA proteins was reduced in the desiccation-induced Suc glassy state. Detailed knowledge about the role of LEA proteins in the stabilization of dry sugar glasses yields information about their role in anhydrobiosis. This may open the possibility to use such proteins in biotechnical applications requiring dry storage of biologicals such as proteins, cells or tissues.

Mechanical characterization of glass fiber resin reinforced with sugarcane

Nowadays, the use of organic fillers in polymer composite materials is on the rise due to ecological concerns. Natural additives are cheap and plentiful in India. Sugarcane is indeed the main staple crop in India. Bagasse is a leftover pellet fiber produced from sugar beet shattered as in the factory. This can be used for the production of biofuels and for certain other purposes. This paper reports the mechanical properties of the manufacturing of epoxy resin fiberglass laminates, including sugar cane and glass fibers mixed with composite resin. Each composite material should be developed and then the step-up produced in line with the ASTM requirements. Each sample will then conduct mechanical properties, including tensile strength, compressive strength and impact strength checking. The findings of the experiment would be correlated between fiber resin laminates and fiber resin blended with sugarcane fiber powder.

Roles of protein, starch and sugar in the texture of sorghum biscuits

This study investigated why biscuits made from sorghum flour have a similar texture to wheat biscuits despite the absence of gluten in sorghum dough. Electron microscopy revealed that the sorghum prolamin protein bodies remained intact in the sorghum biscuits and hence were unlikely to contribute to biscuit structure and texture. Polarized light microscopy showed that the starch granules in the sorghum biscuits were not gelatinized. Increasing dough water content increased the breaking strength and brittleness of sorghum biscuits. However, increasing the proportion of pre-gelatinized sorghum flour in the dough reduced the breaking strength of the sorghum biscuits, indicating that starch gelatinization weakened the biscuit structure. In contrast, increasing the sucrose content of the dough increased sorghum biscuit breaking strength and brittleness. At 20% sucrose (flour basis), the sorghum biscuits had similar breaking strength and brittleness to both Marie and sugar-snap wheat biscuits. DSC and X-ray diffractometry showed that the sugar in both the sorghum and wheat biscuits was in the glassy state and polarized light microscopy revealed that the sugar glass embedded or enveloped the sorghum biscuit flour particles. It is concluded that this sugar glass matrix is responsible for the strength and cohesiveness of the sorghum biscuits.

VaxiPatch™, a novel vaccination system comprised of subunit antigens, adjuvants and microneedle skin delivery: An application to influenza B/Colorado/06/2017

This work introduces VaxiPatch, a novel vaccination system comprised of subunit glycoprotein vaccine antigens, adjuvants and dermal delivery. For this study, rHA of influenza virus B/Colorado/06/2017 was incorporated into synthetic virosomes, and adjuvant liposomes were formed with QS-21 from Saponaria quillaja, with or without the synthetic TLR4 agonist 3D - (6-acyl) PHAD. These components were concentrated and co-formulated into trehalose with dye. Dermal delivery was achieved using an economical 37-point stainless steel microneedle array, designed for automated fill/finish by microfluidic dispensers used for mass production of immunodiagnostics. Vaccine and adjuvant are deposited to form a sugar glass in a pocket on the side of each of the tips, allowing skin penetration to be performed directly by the rigid steel structure. In this study, Sprague Dawley rats (n = 6 per group) were vaccinated by VaxiPatches containing 0.3 µg of rHA, 0.5 µg QS-21 and 0.2 µg 3D - (6-acyl) PHAD and dye, resulting in antigen-specific IgG titers 100-fold higher than 4.5 µg of FluBlok (p = 0.001) delivered intramuscularly. Similarly, hemagglutination inhibition titers in these animals were 14-fold higher than FluBlok controls (p = 0.01). Non-adjuvanted VaxiPatches were also compared with rHA virosomes injected intramuscularly. Accelerated shelf life studies further suggest that formulated virosomal antigens retain activity for at least two months at 60° C. Further, co-formulation of a dye could provide a visible verification of delivery based on the temporary pattern on the skin. A room-temperature-stable vaccination kit such as VaxiPatch has the potential to increase vaccine use and compliance globally.

3D Sugar Printing of Networks Mimicking the Vasculature

The vasculature plays a central role as the highway of the body, through which nutrients and oxygen as well as biochemical factors and signals are distributed by blood flow. Therefore, understanding the flow and distribution of particles inside the vasculature is valuable both in healthy and disease-associated networks. By creating models that mimic the microvasculature fundamental knowledge can be obtained about these parameters. However, microfabrication of such models remains a challenging goal. In this paper we demonstrate a promising 3D sugar printing method that is capable of recapitulating the vascular network geometry with a vessel diameter range of 1 mm down to 150 µm. For this work a dedicated 3D printing setup was built that is capable of accurately printing the sugar glass material with control over fibre diameter and shape. By casting of printed sugar glass networks in PDMS and dissolving the sugar glass, perfusable networks with circular cross-sectional channels are obtained. Using particle image velocimetry, analysis of the flow behaviour was conducted showing a Poisseuille flow profile inside the network and validating the quality of the printing process.

Thermal properties of treated sugar palm yarn/glass fiber reinforced unsaturated polyester hybrid composites

In this work, the effects of alkaline treatment and hybridization on the thermal properties of sugar palm yarn/glass fiber were investigated. The sugar palm fiber was treated with 1% of sodium hydroxide (NaOH) solution for 1h and the ratio of between matrix and reinforcement was 70/30wt.% and 60/40wt.%, respectively, while the ratio of reinforcement between sugar palm yarn fiber and glass fiber was 70/30wt.%, 60/40wt.% and 50/50wt.%, respectively. The thermal properties of the hybrid composites were analyzed using a dynamic mechanical analyzer (DMA) and Thermogravimetric analysis (TGA). The storage modulus (E'), loss modulus (E") and damping factor (tan δ) were evaluated as a function of the alkaline treatment and different percentages of fiber loading. Also, the peak high was investigated for the tan δ curves. In the glassy state area, a higher glass fiber loading hybridized with treated sugar palm fiber exhibited the highest storage modulus, loss modulus and the lowest damping factor. TGA demonstrated that the percentage of residue decreased as the glass fiber loading increased. Overall, the hybridization of glass fiber with treated sugar palm fiber enhances the thermal properties of the hybrid composites for structural applications.

Mechanical properties of sugar palm yarn/woven glass fiber reinforced unsaturated polyester composites: effect of fiber loadings and alkaline treatment

In this paper, hybrid sugar palm yarn and glass fiber reinforced unsaturated polyester composites were investigated in relation to the effects of fiber loadings and alkaline treatment on the composite mechanical properties, such as tensile, flexural, impact and compression strength. The composites were fabricated at a weight ratio of matrix to reinforcement of 70 : 30 and 60 : 40, respectively, while the ratio of sugar palm yarn fiber to glass fiber was selected at 70 : 30, 60 : 40 and 50 : 50, respectively. The results revealed that the mechanical properties of the hybrid composites were increased with an increase of glass fiber loading for both 30 wt % and 40 wt % reinforcement content. The alkaline treatment of the sugar palm fibers have advantageous effect on the hybrid composite performance. The overall results indicated that the developed hybrid composites can be used as an alternative material for glass fiber reinforced polymer composites for various structural applications.

Analysis of dynamic mechanical, low-velocity impact and compression after impact behaviour of benzoyl treated sugar palm/glass/epoxy composites

The objective of this work is to investigate the effect of glass fibre addition and benzoylation treatment on the dynamic mechanical, impact and post-impact properties of sugar palm composites. The hybrid composites were prepared using hand lay-up method with different volume fractions of sugar palm fibre and glass fibre. Dynamic mechanical analysis (DMA) results show that EP/30TSPF/70GF composites exhibited the highest loss and storage modulus compared to EP/UTSPF composites. For the low-velocity impact testing, the experimental results show that the impact energy displayed an excellent relationship with the impact response. The ultrasonic C-scan images show that the damaged area is increasing as the impact energy increasing. From the compression after impact (CAI) studies, it could be resolved that the impact damage exhibited outstanding connection with the impact response and compressive strength of the EP/30TSPF/70GF composite decreased as the impact energy increasing. Overall, it can be concluded that the benzoylation treatment and glass fibre addition on the sugar palm composites enhances the dynamic mechanical, impact and post-impact properties of sugar palm composites.

Thermostable lyoprotectant-enhanced cell-free protein synthesis for on-demand endotoxin-free therapeutic production

Cell-free protein synthesis has emerged as a promising platform for the production of therapeutic proteins due to its inherently open reaction environment, flexible reaction conditions and rapid protein synthesis capabilities. In recent years, lyophilized cell-free systems have widened the application space of cell-free technology by improving reagent stability outside of cold-chain storage. Current embodiments of the system, however, demonstrate poor stability at elevated temperatures. Lyoprotectants have long been recognized for the ability to preserve the activity of biological molecules during drying processes, but the application of this technology to lyophilized cell-free systems has been limited and has failed to address the negative effects that such lyoprotectants may have on cell-free systems. Here, several lyoprotected, lyophilized cell-free protein synthesis systems are demonstrated using antiplasticized sugar glasses as lyoprotectants, showing significant improvement over standard lyophilized systems or trehalose-preserved systems. Furthermore, we demonstrate for the first time, preservation and therapeutic expression, specifically of FDA-approved crisantaspase, from a truly single-pot lyophilized, endotoxin-free, cell-free protein synthesis system, exemplifying the potential for on-site therapeutic synthesis.

Использование сиропов в рецептуре сахаристых кондитерских изделий повышенной пищевой ценности

Confectionery products of high nutritional value are affordable and rich in vitamins and minerals. Their development requires new raw materials with a high physiological value, which would be available on the local consumer market. Vegetable syrups can be used in sugar confectionery products, especially if they are fortified with vitamins and minerals. The present research proves the expediency of using fortified syrups in the production of sugar glasses. This type of caramel has a high energy value but no biologically valuable substances in the composition. Therefore, fortified syrups increase the biological value of the product. The study featured samples of sugar glasses with syrups fortified with rosehip, sea buckthorn, lemongrass, and cranberries. The paper describes their nutritional value, as well as the therapeutic and prophylactic properties of the syrups. It also contains data on the sensory quality, physic-chemical parameters, and the nutritional value.