Effects of aftercrop tomato and maize on the soil microenvironment and microbial diversity in a long-term cotton continuous cropping field.

Long-term continuous cropping affects the soil microecological community and leads to nutrient imbalances, which reduces crop yields, and crop rotation can increase soil productivity. To study the effects of the cultivation of tomato (Solanum lycopersicum) and corn (Zea mays) on the microbial community, physical and chemical factors and the structure of aggregates in cotton (Gossypium hirsutum) long-term continuous cropping soils were examined. Four cropping patterns were established, including one continuous cropping pattern and three crop rotation patterns, and the diversity of the soil microecological community was measured using high-throughput sequencing. The physical and chemical properties of different models of soil were measured, and the soil aggregate structure was determined by dry and wet sieving. Planting of aftercrop tomato and corn altered the bacterial community of the cotton continuous soil to a lesser extent and the fungal community to a greater extent. In addition, continuous cropping reduced the diversity and richness of the soil fungal community. Different aftercrop planting patterns showed that there were very high contents of soil organic carbon and organic matter in the cotton-maize rotation model, while the soil aggregate structure was the most stable in the corn-cotton rotation model. Planting tomato in continuous cropping cotton fields has a greater effect on the soil microbial community than planting maize. Therefore, according to the characteristics of different succeeding crop planting patterns, the damage of continuous cropping of cotton to the soil microenvironment can be alleviated directionally, which will enable the sustainable development of cotton production.

Effect of Aminating Lignin Loading with Arbuscular Mycorrhizal Fungi on Soil Aggregate Structure Improvement.

Lignin is an important component of plant fiber raw materials, and is a three-dimensional network structure aromatic polymer with abundant resources and a complex structure in nature. Lignin is generally used as industrial waste, and its potential value has not been fully utilized. Modern agriculture extensively uses chemical fertilizers, leading to the gradual degradation of soil fertility and structure, which seriously affects crop growth, nutrient transport, and root respiration function. Based on soil bulk density, porosity, aggregates, and their stability indicators, this study analyzed the effects of aminated industrial lignin and its loading with arbuscular mycorrhizal fungi on soil structure improvement and plant growth. It was hoped that resource-rich lignin could play a beneficial role in improving soil structure and promoting crop growth. The phenolic hydroxyl group of lignin was epoxidized and further aminated to load with arbuscular mycorrhizal fungi. The results indicated that amine-modified lignin could effectively load with arbuscular mycorrhizal fungi. The application of arbuscular mycorrhizal fungi-supported aminated lignin to soil aggregate structure improvement greatly reduced the bulk density of soil, and increased the porosity of soil and the content of large granular soil. Compared with unmodified soil, soil bulk density decreased by 73.08%, the porosity of soil increased by 70.43%, and the content of large granular soil increased by 56.38%. Using the improved soil for corn cultivation efficiently increased the biomass of corn. The plant height was increased by 72.16%, the root-shoot ratio was increased by 156.25%, and other indexes were also improved to varying degrees. The experimental method provides an important basis for the effective utilization of lignin materials in agriculture in the future.

Impact of flaxseed gum on the aggregate structure, pasting properties, and rheological behavior of waxy rice starch.

This study examines the effects of flaxseed gum (FG) on the aggregate structure, pasting and rheological properties of waxy rice starch (WRS). Results display an increase in the ordered molecular structure (R1047/1024), relative crystallinity (RC), compactness (α), and microphase heterogeneity (ε, density degree of nanoaggregates, from 3.52 to 4.23) for WRS-FG complexes. These suggested FG facilitated the development of more organized molecular and crystalline structures of WRS, accompanied by the formation of ordered nanoaggregates with higher density (i.e., nano-aggregation structure). Also, FG addition resulted in the formation of enhanced gel network structure characterized by thicker layer walls and more uniform pores. These structural transformations contributed to a rise in gelatinization temperature (To, from 56.90 °C to 62.10 °C) and enthalpy (ΔH), as well as alterations in paste viscosities (PV, from 1285.00 mPa·s to 1734.00 mPa·s), and the rigidity of network structure (e.g., decreased loss tangent). These results indicate that FG could effectively regulate the techno-functional properties of WRS by rationally controlling the starch intrinsic structures of starch. And this study may improve the pasting and gelling properties of starch, thus driving the development of high-quality starchy foods and prolonging their shelf life, especially for glutinous rice flour products.

Growth prolongation of human induced pluripotent stem cell aggregate in three-dimensional suspension culture system by addition of botulinum hemagglutinin.

Human induced pluripotent stem cells (hiPSCs) can be used in regenerative therapy as an irresistible cell source, and so the development of scalable production of hiPSCs for three-dimensional (3D) suspension culture is required. In this study, we established a simple culture strategy for improving hiPSC aggregate growth using botulinum hemagglutinin (HA), which disrupts cell-cell adhesion mediated by E-cadherin. When HA was added to the suspension culture of hiPSC aggregates, E-cadherin-mediated cell-cell adhesion was temporarily disrupted within 24 h, but then recovered. Phosphorylated myosin light chain, a contractile force marker, was also recovered at the periphery of hiPSC aggregates. The cell aggregates were suppressed the formation of collagen type I shell-like structures at the periphery by HA and collagen type I was homogenously distributed within the cell aggregates. In addition, these cell aggregates retained the proliferation marker Ki-67 throughout the cell aggregates. The apparent specific growth rate with HA addition was maintained continuously throughout the culture, and the final cell density was 1.7-fold higher than that in the control culture. These cells retained high expression levels of pluripotency markers. These observations indicated that relaxation of cell-cell adhesions by HA addition induced rearrangement of the mechanical tensions generated by actomyosin in hiPSC aggregates and suppression of collagen type I shell-like structure formation. These results suggest that this simple and readily culture strategy is a potentially useful tool for improving the scalable production of hiPSCs for 3D suspension cultures.

Mesoscopic Mechanical Properties of Aggregate Structure in Asphalt Mixtures and Gradation Optimization.

Particle media are widely used in engineering and greatly influence the performance of engineering materials. Asphalt mixtures are multi-phase composite materials, of which coarse aggregates account for more than 60%. These coarse aggregates form a stable structure to transfer and disperse traffic loads. Therefore, knowing how to adjust the structural composition of coarse aggregates to optimize their performance is the key to optimize the performance of asphalt mixtures. In this study, the effects of different roughness and different sizes on the interlocking force and contact force of coarse aggregates were investigated through means of simulation (DEM), and then the formation-evolution mechanism of the coarse aggregate structure and the role of different sizes of aggregates in the coarse aggregate structure were analyzed. Subsequently, the optimal ratio of coarse aggregates was explored through indoor tests, and finally, the gradation of asphalt mixture based on the optimization of fine structure was formed and verified through indoor tests. The results showed that the major model can effectively reveal the role of different types of aggregates in the fine structure and the relationship between the strength of contact forces between them and clarify that the strength of the fine structure increases with the increase in aggregate roughness. Hence, the coarse aggregate structure can be regarded as a contact force transmission system composed of some strong and sub-strong contact forces. Their formation-evolution mechanism can be regarded as a process of the formation of strong and sub-strong contact forces and the transformation from sub-strong contact force to strong contact force. Moreover, the dynamic stability of the optimized graded asphalt mixture was increased by 30%, and the fracture toughness was increased by 26%.

Full-length prion protein incorporated into prion aggregates is a marker for prion strain-specific destabilization of aggregate structure following cellular uptake.

Accumulation of insoluble aggregates of infectious, partially protease-resistant prion protein (PrPD) generated via the misfolding of protease sensitive prion protein (PrPC) into the same infectious conformer, is a hallmark of prion diseases. Aggregated PrPD is taken up and degraded by cells, a process likely involving changes in aggregate structure that can be monitored by accessibility of the N-terminus of full-length PrPD to cellular proteases. We therefore tracked the protease sensitivity of full-length PrPD before and after cellular uptake for two murine prion strains, 22L and 87V. For both strains, PrPD aggregates were less stable following cellular uptake with increased accessibility of the N-terminus to cellular proteases across most aggregate sizes. However, a limited size range of aggregates was able to better protect the N-termini of full-length PrPD, with the N-terminus of 22L-derived PrPD more protected than that of 87V. Interestingly, changes in aggregate structure were associated with minimal changes to the protease-resistant core of PrPD. Our data show that cells destabilize the aggregate quaternary structure protecting PrPD from proteases in a strain-dependent manner, with structural changes exposing protease sensitive PrPD having little effect on the protease-resistant core, and thus conformation, of aggregated PrPD.

Amidoamine Oxide Surfactants as Low-Molecular-Weight Hydrogelators: Effect of Methylene Chain Length on Aggregate Structure and Rheological Behavior.

Rheology control is an important issue in many industrial products such as cosmetics and paints. Recently, low-molecular-weight compounds have attracted considerable attention as thickeners/gelators for various solvents; however, there is still a significant need for molecular design guidelines for industrial applications. Amidoamine oxides (AAOs), which are long-chain alkylamine oxides with three amide groups, are surfactants that act as hydrogelators. Here, we show the relationship between the length of methylene chains at four different locations of AAOs, the aggregate structure, the gelation temperature Tgel, and the viscoelasticity of the formed hydrogels. As seen from the results of electron microscopic observations, the aggregate structure (ribbon-like or rod-like) can be controlled by changing the length of methylene chain in the hydrophobic part, the length of methylene chain between the amide and amine oxide groups, and the lengths of methylene chains between amide groups. Furthermore, hydrogels consisting of rod-like aggregates showed significantly higher viscoelasticity than those consisting of ribbon-like aggregates. In other words, it was shown that the gel viscoelasticity could be controlled by changing the methylene chain lengths at four different locations of the AAO.

Mutant SOD1 aggregates formed in vitro and in cultured cells are polymorphic and differ from those arising in the CNS.

Mutations in the human Superoxide dismutase 1 (hSOD1) gene are well-established cause of the motor neuron disease ALS. Patients and transgenic (Tg) ALS model mice carrying mutant variants develop hSOD1 aggregates in the CNS. We have identified two hSOD1 aggregate strains, which both transmit spreading template-directed aggregation and premature fatal paralysis when inoculated into adult transgenic mice. This prion-like spread of aggregation could be a primary disease mechanism in SOD1-induced ALS. Human SOD1 aggregation has been studied extensively both in cultured cells and under various conditions in vitro. To determine how the structure of aggregates formed in these model systems related to disease-associated aggregates in the CNS, we used a binary epitope-mapping assay to examine aggregates of hSOD1 variants G93A, G85R, A4V, D90A, and G127X formed in vitro, in four different cell lines and in the CNS of Tg mice. We found considerable variability between replicate sets of in vitro-generated aggregates. In contrast, there was a high similarity between replicates of a given hSOD1 mutant in a given cell line, but pronounced variations between different hSOD1 mutants and different cell lines in both structures and amounts of aggregates formed. The aggregates formed in vitro or in cultured cells did not replicate the aggregate strains that arise in the CNS. Our findings suggest that the distinct aggregate morphologies in the CNS could result from a micro-environment with stringent quality control combined with second-order selection by spreading ability. Explorations of pathogenesis and development of therapeutics should be conducted in models that replicate aggregate structures forming in the CNS.

Immature ALS-associated mutant superoxide dismutases form variable aggregate structures through distinct oligomerization processes.

Protein misfolding and aggregation are hallmarks of many diseases, including amyotrophic lateral sclerosis (ALS). In familial ALS, aberrant self-association of mutant Cu,Zn-superoxide dismutase (SOD1) is implicated as a key contributor to disease. Mutations have the largest impacts on the stability of the most immature form of SOD1, the unmetallated, disulfide-reduced monomer (apoSH SOD1). Here we demonstrate that, despite the marginal stability of apoSH SOD1, aggregation is little correlated with the degree of protein unfolding, and multiple modes of aggregation occur, depending on the mutation and solution conditions. Light scattering and atomic force microscopy reveal two distinct mutant SOD1 behaviours: high aggregator mutants form abundant small assemblies, while low aggregator mutants form fewer, more fibre-like aggregates. Attenuated total reflectance-Fourier transform infrared spectroscopy and Thioflavin T binding show the aggregates maintain native-like anti-parallel beta structure. These results provide new evidence that ALS-associated mutations promote the aggregation of apoSH SOD1 through multiple pathways, with broad implications for understanding mechanisms of protein self-association in disease and biotechnology.

Methodological advances and strategies for high resolution structure determination of cellular protein aggregates.

Aggregation of proteins is at the nexus of molecular processes crucial to aging, disease, and employing proteins for biotechnology and medical applications. There has been much recent progress in determining the structural features of protein aggregates that form in cells; yet, owing to prevalent heterogeneity in aggregation, many aspects remain obscure and often experimentally intractable to define. Here, we review recent results of structural studies for cell-derived aggregates of normally globular proteins, with a focus on high-resolution methods for their analysis and prediction. Complementary results obtained by solid-state NMR spectroscopy, FTIR spectroscopy and microspectroscopy, cryo-EM, and amide hydrogen/deuterium exchange measured by NMR and mass spectrometry, applied to bacterial inclusion bodies and disease inclusions, are uncovering novel information on in-cell aggregation patterns as well as great diversity in the structural features of useful and aberrant protein aggregates. Using these advances as a guide, this review aims to advise the reader on which combination of approaches may be the most appropriate to apply to their unique system.

Quenched hydrogen-deuterium amide exchange optimization for high-resolution structural analysis of cellular protein aggregates.

Inclusion bodies (IBs) are large, insoluble aggregates that often form during the overexpression of proteins in bacteria. These aggregates are of broad fundamental and practical significance, for recombinant protein preparation and due to their relevance to aggregation-related medical conditions and their recent emergence as promising functional nanomaterials. Despite their significance, high resolution knowledge of IB structure remains very limited. Such knowledge will advance understanding and control of IB formation and properties in myriad practical applications. Here, we report a detailed quenched hydrogen-deuterium amide exchange (qHDX) method with NMR readout to define the structure of IBs at the level of individual residues throughout the protein. Applying proper control of experimental conditions, such as sample pH, water content, temperature, and intrinsic rate of amide exchange, yields in depth results for these cellular protein aggregates. qHDX results illustrated for Cu, Zn superoxide dismutase 1 (SOD1) and Adnectins show their IBs include native-like structure and some but not all mutations alter IB structure.

Electrical impedance spectroscopy as a potential tool to investigate the structure and size of aggregates during water and wastewater treatment.

Microscopic structure and size are important metrics for estimating aggregates environmental behaviors during water and wastewater treatment. However, in-situ determination of these characteristics is still a challenge. Here, we drew inspiration from a block disassembly process to propose an electrical impedance spectroscopy (EIS) method and constructed a generalized framework to associate macroscale electrical properties with microscopic structure and size-related characteristics of aggregates of different hierarchies. Extracted via EIS, the proposed models were verified to be capable of describing the self-similarity of aggregates and capturing the fractal and size information. Further, the proposed models exhibited a wide range of applications, which agrees well with the data gathered from various activated sludges, other colloids, and microgels in water and wastewater treatment. Finally, the EIS method was achieved online monitoring of fractal dimension and floc size during a sludge pre-oxidation conditioning process, which was elected as an example to illustrate the potential online applications of this EIS method in real water and wastewater environment. The obtained on-line data were used to indicate the potential suitable oxidation time during sludge pre-oxidation conditioning. These observations may inspire new methods of quantifying the aggregate structure and promote intelligent and dynamic decision-making during water and wastewater treatment.

Conversion of alpine pastureland to artificial grassland altered CO2 and N2O emissions by decreasing C and N in different soil aggregates.

BACKGROUND: The impacts of land use on greenhouse gases (GHGs) emissions have been extensively studied. However, the underlying mechanisms on how soil aggregate structure, soil organic carbon (SOC) and total N (TN) distributions in different soil aggregate sizes influencing carbon dioxide (CO2), and nitrous oxide (N2O) emissions from alpine grassland ecosystems remain largely unexplored. METHODS: A microcosm experiment was conducted to investigate the effect of land use change on CO2and N2O emissions from different soil aggregate fractions. Soil samples were collected from three land use types, i.e., non-grazing natural grassland (CK), grazing grassland (GG), and artificial grassland (GC) in the Bayinbuluk alpine pastureland. Soil aggregate fractionation was performed using a wet-sieving method. The variations of soil aggregate structure, SOC, and TN in different soil aggregates were measured. The fluxes of CO2 and N2O were measured by a gas chromatograph. RESULTS: Compared to CK and GG, GC treatment significantly decreased SOC (by 24.9-45.2%) and TN (by 20.6-41.6%) across all soil aggregate sizes, and altered their distributions among soil aggregate fractions. The cumulative emissions of CO2 and N2O in soil aggregate fractions in the treatments of CK and GG were 39.5-76.1% and 92.7-96.7% higher than in the GC treatment, respectively. Moreover, cumulative CO2emissions from different soil aggregate sizes in the treatments of CK and GG followed the order of small macroaggregates (2-0.25 mm) > large macroaggregates (> 2 mm) > micro aggregates (0.25-0.053 mm) > clay +silt (< 0.053 mm), whereas it decreased with aggregate sizes decreasing in the GC treatment. Additionally, soil CO2 emissions were positively correlated with SOC and TN contents. The highest cumulative N2O emission occurred in micro aggregates under the treatments of CK and GG, and N2O emissions among different aggregate sizes almost no significant difference under the GC treatment. CONCLUSIONS: Conversion of natural grassland to artificial grassland changed the pattern of CO2 emissions from different soil aggregate fractions by deteriorating soil aggregate structure and altering soil SOC and TN distributions. Our findings will be helpful to develop a pragmatic management strategy for mitigating GHGs emissions from alpine grassland.

Characterization of silver nanoparticle aggregates using single particle-inductively coupled plasma-mass spectrometry (spICP-MS).

The single particle-inductively coupled plasma-mass spectrometry was applied to characterize the aggregates of AgNPs. was applied to characterize the aggregates of AgNPs. Two sizes of citrate-AgNPs and PVP-AgNPs were used at relatively high and predicted environmental concentrations under various ionic strengths. Citrate-AgNP aggregated with increases in the ionic strength, whereas PVP-AgNPs were sterically stable. The critical coagulation concentrations were 85 mM and 100 mM NaNO3 for 60 nm and 100 nm citrate-AgNPs at 2 mg L-1 as total Ag obtained by dynamic light scattering (DLS). At 2 mg L-1 as total Ag, the mass of an aggregate gradually increased with increasing ionic strength for both citrate-AgNP during spICP-MS analyses. The average number of single particles derived from the mass in an aggregate was calculated to be 8.68 and 5.95 for 60 nm and 100 nm citrate-AgNPs at 85 mM and 100 mM NaNO3, respectively after 2 h. The mass fractal dimensions were determined to be 2.97 and 2.83, further implying that the aggregate structures were very rigid and compact. Only marginal increases in the average mass and number of single particles in the aggregate units were found during 24 h under environmentally relevant AgNP concentrations. The average number of single particles constituting an aggregate unit for 60 nm and 100 nm citrate-AgNPs was 1.24 and 1.37 after 24 h at a high ionic strength. These results indicate that under environmentally relevant conditions, the collision frequency is predominant in the aggregation and that NPs are likely to encounter natural colloids such as clay and organic matter to form hetero-aggregates.

Hierarchical Structure Formation of Nanoparticulate Spray-Dried Composite Aggregates.

The design of hierarchically structured nano- and microparticles of different sizes, porosities, surface areas, compositions, and internal structures from nanoparticle building blocks is important for new or enhanced application properties of high-quality products in a variety of industries. Spray-drying processes are well-suited for the design of hierarchical structures of multicomponent products. This structure design using various nanoparticles as building blocks is one of the most important challenges for the future to create products with optimized or completely new properties. Furthermore, the transfer of designed nanomaterials to large-scale products with favorable handling and processing can be achieved. The resultant aggregate structure depends on the utilized nanoparticle building blocks as well as on a large number of process and formulation parameters. In this study, structure formation and segregation phenomena during the spray drying process were investigated to enable the synthesis of tailor-made nanostructures with defined properties. Moreover, a theoretical model of this segregation and structure formation in nanosuspensions is presented using a discrete element method simulation.

Kinetic modeling of AS(III) and AS(V) adsorption by a novel tetravalent manganese feroxyhyte.

HYPOTHESIS: The purpose of the present work is the development of a kinetic model for the adsorption of As(III) and As(V) onto tetravalent manganese feroxyhyte (δ-Fe0.75Mn0.25OOH), which have been recently proved to be very efficient adsorbent for the particular species. EXPERIMENTS: In this respect equilibrium and adsorption kinetic experiments onto this type of adsorbent for As(III) and As(V) were performed. Two sizes of adsorbate particles are tested in order to acquire better insight to the adsorption process. RESULTS: The adsorption kinetic curves cannot be described by the well-known adsorption kinetic models so a detailed model that takes into account the structure of the adsorbent particle is developed. The model parameters were extracted by the requirement of agreement between model and experimental results. The batch model developed here is necessary for the development of models for fixed bed adsorption devices in order to exploit the commercial prospects of the particular adsorbent. This work constitutes the first attempt of kinetic study and adsorption model development for the specific very promising adsorbent.

Agglomeration behaviour of titanium dioxide nanoparticles in river waters: A multi-method approach combining light scattering and field-flow fractionation techniques.

Titanium dioxide nanoparticles (TiO2 NPs) are currently one of the most prolifically used nanomaterials, resulting in an increasing likelihood of release to the environment. This is of concern as the potential toxicity of TiO2 NPs has been investigated in several recent studies. Research into their fate and behaviour once entering the environment is urgently needed to support risk assessment and policy development. In this study, we used a multi-method approach combining light scattering and field-flow fractionation techniques to assess both the aggregation behaviour and aggregate structure of TiO2 NPs in different river waters. Results showed that both the aggregate size and surface-adsorbed dissolved organic matter (DOM) were strongly related to the initial DOM concentration of the tested waters (i.e. R(2) > 0.90) suggesting that aggregation of TiO2 NPs is controlled by the presence and concentration of DOM. The conformation of the formed aggregates was also found to be strongly related to the surface-adsorbed DOM (i.e. R(2) > 0.95) with increasing surface-adsorbed DOM leading to more compact structures. Finally, the concentration of TiO2 NPs remaining in the supernatant after sedimentation of the larger aggregates was found to decrease proportionally with both increasing IS and decreasing DOM concentration, resulting in more than 95% sedimentation in the highest IS sample.

Aggregation behaviour of engineered nanoparticles in natural waters: characterising aggregate structure using on-line laser light scattering.

Adsorption of natural organic matter, aggregation and disaggregation have been identified as three of the main processes affecting the fate and behaviour of engineered nanoparticles (ENPs) in aquatic environments. However, although several methods have been developed to study the aggregation behaviour of ENPs in natural waters, there are only a few studies focusing on the fate of such aggregates and their potential disaggregation behaviour. In this study, we proposed and demonstrated a simple method for characterising the aggregation behaviour and aggregate structure of ENPs in different natural waters. Both the aggregate size of ENPs and their adsorption capacity for dissolved organic matter (DOM) were strongly related (R(2)>0.97, p<.05) to the combined effect of initial concentration of dissolved organic matter (DOM) and the ionic strength of the natural waters. The structure of the formed aggregates was strongly correlated (R(2)>0.95, p<.05) to the amount of DOM adsorbed by the ENPs during the aggregation process. Under high ionic strength conditions, aggregation is mainly governed by diffusion and the aggregates formed under these conditions showed the lowest stability and fractal dimension, forming linear, chain-like aggregates. In contrast, under low ionic strength conditions, the aggregate structure was more compact, most likely due to strong chemical binding with DOM and bridging mechanisms involving divalent cations formed during reaction-limited aggregation.

Mechanism of Hbγ-35-induced an increase in the activation of the human immune system by endotoxins.

Endotoxins (LPS) are highly potent immune stimulatory molecules and are mainly known for triggering Gram-negative sepsis. However, besides their toxic effects, this stimulatory function may be advantageous, for example when used as an adjuvant during vaccination. Thus, there is always a narrow range between the useful wake-up of the immune system and its overwhelming reaction, which can lead to diseases like sepsis. This raises the question of which conformational properties are responsible for making the LPS aggregates more or less potent. As described previously, the size, type and form of LPS aggregates play a major role in their immune stimulatory activity. In this study we investigate the role of these parameters. On the one hand, we use a peptide (Pep19-2.5; Aspidasept) that causes a change of the LPS aggregate structure into a less toxic state; on the other hand, we use a potent immune stimulating peptide (Hbγ-35), leading to higher toxicity. We have found opposing effects on LPS aggregate conformations allowing a better understanding of the processes of immune stimulation.

Reduction of the C191-C220 disulfide of α-chymotrypsinogen A reduces nucleation barriers for aggregation.

Proper disulfide formation can be essential for the conformational stability of natively folded proteins. For proteins that must unfold in order to aggregate, disruption of native disulfides may therefore promote aggregation. This study characterizes differences in the aggregation process for wild-type (WT) α-chymostrypsinogen A (aCgn) and the same molecule with one of its native disulfides (C191-C220) reduced to free thiols (aCgnSH) at acidic pH, where WT aCgn forms semi-flexible amyloid polymers. Loss of the disulfide leads to no discernable differences in folded monomer secondary or tertiary structure based on circular dichroism (CD) or intrinsic fluorescence (FL), and causes a small decrease in the free energy change upon unfolding. After unfolding-mediated aggregation, the resulting amyloid morphology and structure are similar or indistinguishable for aCgn and aCgnSH by CD, FL, ThT binding, multi-angle laser light scattering, and transmission electron microscopy. Aggregates of aCgn and aCgnSH are also able to cross-seed with monomers of the other species. However, aggregates of aCgnSH are more resistive than aCgn aggregates to urea-mediated dissociation, suggesting some degree of structural differences in the aggregated species that was not resolvable in detail without higher resolution methods. Mechanistic analyses of aggregation kinetics indicate that the initiation or nucleation of new aggregates from aCgnSH involves a mono-molecular rate limiting step, possibly the unfolding step. In contrast, that for aCgn involves an oligomeric intermediate, suggesting native disulfide linkages help to hinder non-native protein aggregation by providing conformational barriers to key nucleation event(s).