Buffering capacity is determinant for restoring early α-synuclein aggregation

2022 ◽  
pp. 106760
Author(s):  
Marco A. Saraiva ◽  
M. Helena Florêncio
Keyword(s):  
1998 ◽  
Vol 38 (4-5) ◽  
pp. 29-35 ◽  
Author(s):  
C. J. Banks ◽  
P. N. Humphreys

The stability and operational performance of single stage digestion with and without liquor recycle and two stage digestion were assessed using a mixture of paper and wood as the digestion substrate. Attempts to maintain stable digestion in both single stage reactors were unsuccessful due to the inherently low natural buffering capacity exhibited; this resulted in a rapid souring of the reactor due to unbuffered volatile fatty acid (VFA) accumulation. The use of lime to control pH was unsatisfactory due to interference with the carbonate/bicarbonate equilibrium resulting in wide oscillations in the control parameter. The two stage system overcame the pH stability problems allowing stable operation for a period of 200 days without any requirement for pH control; this was attributed to the rapid flushing of VFA from the first stage reactor into the second stage, where efficient conversion to methane was established. Reactor performance was judged to be satisfactory with the breakdown of 53% of influent volatile solids. It was concluded that the reactor configuration of the two stage system offers the potential for the treatment of cellulosic wastes with a sub-optimal carbon to nitrogen ratio for conventional digestion.


Pharmaceutics ◽  
2021 ◽  
Vol 13 (1) ◽  
pp. 87
Author(s):  
Jaehong Park ◽  
Kyusik Kim ◽  
Sohee Jeong ◽  
Migyeom Lee ◽  
Tae-il Kim

In this work, highly osmotic oxidized sucrose-crosslinked polyethylenimine (SP2K) polymers were developed for gene delivery systems, and the transfection mechanism is examined. First, periodate-oxidized sucrose and polyethylenimine 2K (PEI2K) were crosslinked with various feed ratios via reductive amination. The synthesis was confirmed by 1H NMR and FTIR. The synthesized SP2K polymers could form positively charged (~40 mV zeta-potential) and nano-sized (150–200 nm) spherical polyplexes with plasmid DNA (pDNA). They showed lower cytotoxicity than PEI25K but concentration-dependent cytotoxicity. Among them, SP2K7 and SP2K10 showed higher transfection efficiency than PEI25K in both serum and serum-free conditions, revealing the good serum stability. It was found that SP2K polymers possessed high osmolality and endosome buffering capacity. The transfection experiments with cellular uptake inhibitors suggest that the transfection of SP2K polymers would progress by multiple pathways, including caveolae-mediated endocytosis. It was also thought that caveolae-mediated endocytosis of SP2K polyplexes would be facilitated through cyclooxygenase-2 (COX-2) expression induced by high osmotic pressure of SP2K polymers. Confocal microscopy results also supported that SP2K polyplexes would be internalized into cells via multiple pathways and escape endosomes efficiently via high osmolality and endosome buffering capacity. These results demonstrate the potential of SP2K polymers for gene delivery systems.


2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Rianne C. Prins ◽  
Sonja Billerbeck

Abstract Background Fungi are premier hosts for the high-yield secretion of proteins for biomedical and industrial applications. The stability and activity of these secreted proteins is often dependent on the culture pH. As yeast acidifies the commonly used synthetic complete drop-out (SD) media that contains ammonium sulfate, the pH of the media needs to be buffered in order to maintain a desired extracellular pH during biomass production. At the same time, many buffering agents affect growth at the concentrations needed to support a stable pH. Although the standard for biotechnological research and development is shaken batch cultures or microtiter plate cultures that cannot be easily automatically pH-adjusted during growth, there is no comparative study that evaluates the buffering capacity and growth effects of different media types across pH-values in order to develop a pH-stable batch culture system. Results We systematically test the buffering capacity and growth effects of a citrate-phosphate buffer (CPB) from acidic to neutral pH across different media types. These media types differ in their nitrogen source (ammonium sulfate, urea or both). We find that the widely used synthetic drop-out media that uses ammonium sulfate as nitrogen source can only be effectively buffered at buffer concentrations that also affect growth. At lower concentrations, yeast biomass production still acidifies the media. When replacing the ammonium sulfate with urea, the media alkalizes. We then develop a medium combining ammonium sulfate and urea which can be buffered at low CPB concentrations that do not affect growth. In addition, we show that a buffer based on Tris/HCl is not effective in maintaining any of our media types at neutral pH even at relatively high concentrations. Conclusion Here we show that the buffering of yeast batch cultures is not straight-forward and addition of a buffering agent to set a desired starting pH does not guarantee pH-maintenance during growth. In response, we present a buffered media system based on an ammonium sulfate/urea medium that enables relatively stable pH-maintenance across a wide pH-range without affecting growth. This buffering system is useful for protein-secretion-screenings, antifungal activity assays, as well as for other pH-dependent basic biology or biotechnology projects.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Yi Wen ◽  
Nazila Salamat-Miller ◽  
Keethkumar Jain ◽  
Katherine Taylor

AbstractDirect delivery of therapeutic enzymes to the Central Nervous System requires stringent formulation design. Not only should the formulation design consider the delicate balance of existing ions, proteins, and osmolality in the cerebrospinal fluid, it must also provide long term efficacy and stability for the enzyme. One fundamental approach to this predicament is designing formulations with no buffering species. In this study, we report a high concentration, saline-based formulation for a human sulfatase for its delivery into the intrathecal space. A high concentration formulation (≤ 40 mg/mL) was developed through a series of systematic studies that demonstrated the feasibility of a self-buffered formulation for this molecule. The self-buffering capacity phenomenon was found to be a product of both the protein itself and potentially the residual phosphates associated with the protein. To date, the self-buffered formulation for this molecule has been stable for up to 4 years when stored at 5 ± 3 °C, with no changes either in the pH values or other quality attributes of the molecule. The high concentration self-buffered protein formulation was also observed to be stable when exposed to multiple freeze–thaw cycles and was robust during in-use and agitation studies.


Agronomy ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 851
Author(s):  
Sonia Cacini ◽  
Sara Di Lonardo ◽  
Simone Orsenigo ◽  
Daniele Massa

Professional peat-free substrates for ornamental plant production are increasingly required by nursery growers. Most promising materials are green compost, coconut coir dust, and woody fibre, used alone or in mixtures. One of the major concerns is pH, usually higher than optimal. In this work, a method based on a three-step procedure was adopted to acidify three organic matrices alone or in mixtures and to individuate the most suitable product, between iron(II) sulphate 7-hydrate and elemental sulphur chips. Firstly, the determination of the buffering capacity by dilution with sulphuric acid was carried out to determine dosages. Afterwards, an incubation trial of 84 (iron(II) sulphate) or 120 days (sulphur chips) was conducted on matrices and substrate mixtures with calculated doses in a climatic chamber maintained at 21 °C. Iron(II) sulphate resulted not suitable because it caused a rapid, but not lasting, pH lowering and an excessive electrical conductivity (EC) increase. Sulphur chips could instead guarantee an adequate and lasting pH lowering. These results were then validated in the open field trial on matrices and substrates. The proposed acidification methodology could be considered in developing new substrates, but the rapidity of pH acidification and EC increase on plant and mineral nutrition should be further investigated.


2021 ◽  
Author(s):  
Anirban Akhand ◽  
Abhra Chanda ◽  
Kenta Watanabe ◽  
Sourav Das ◽  
Tatsuki Tokoro ◽  
...  

AbstractGlobally, water bodies adjacent to mangroves are considered significant sources of atmospheric CO2. We directly measured the partial pressure of CO2 in water [pCO2(water)] and related biogeochemical parameters with high temporal resolution, covering both diel and tidal cycles, in the mangrove-surrounding waters around the northern Bay of Bengal during the post-monsoon season. Mean pCO2(water) was marginally oversaturated in two creeks (470 ± 162 µatm, mean ± SD) and undersaturated in the adjoining estuarine stations (387 ± 58 µatm) compared to atmospheric pCO2, and was considerably lower than the global average. We further estimated the pCO2(water) and buffering capacity of all possible sources of the mangrove-surrounding waters and concluded that their character as a CO2 sink or weak source is due to the predominance of marine water from the Bay of Bengal with low pCO2 and high buffering capacity. Marine water with high buffering capacity suppresses the effect of pCO2 increase within the mangrove system and lowers the CO2 evasion even in creek stations. The δ13C of dissolved inorganic carbon (DIC) in the mangrove-surrounding waters indicated that the DIC sources were a mixture of mangrove plants, pore-water, and groundwater, in addition to marine water. Finally, we showed that the CO2 evasion rate from the estuaries of the Sundarbans is much lower than the recently estimated world average. Our results demonstrate that mangrove areas having such low emissions should be considered when up-scaling the global mangrove carbon budget from regional observations.


1970 ◽  
Vol 75 (3) ◽  
pp. 517-521 ◽  
Author(s):  
D. I. H. Jones

SUMMARYThe effect of three levels of N fertilizer on the ensiling characteristics of S. 24 perennial ryegrass and S. 37 cocksfoot have been examined during first growth in two growing seasons. The effects of sucrose supplementation, inoculation with Lactobacillus plantarum and wilting were also examined in certain cuts. All silages were made in the laboratory using a small scale vacuum silage technique.The perennial ryegrass herbage was higher in water soluble carbohydrates than the cocksfoot, N fertilizers decreased soluble carbohydrates and dry-matter content in both species. Buffering capacity was not consistently different between grasses or between N levels.Herbage was cut at two stages of maturity in the first year. In the first cut (8 days before ear emergence), perennial ryegrass silages were well preserved irrespective of the amount of N applied to the grass. Cocksfoot silages were well preserved only when the lowest level of N fertilizer had been applied (50 kg/ha). Supplementation of cocksfoot with sucrose prior to ensiling markedly improved silage quality, but inoculation had no effect. In the second cut (26 days after ear emergence) the grasses were higher in drymatter content and showed a lower buffering capacity, but neither ryegrass nor cocksfoot silages were well preserved unless supplemented with sucrose prior to ensiling.In the second year of the experiment only one cut was taken (9 days after ear emergence). As in the previous year, silages made from herbage at a late stage of growth were poorly preserved. Wilting prior to ensiling resulted in well-preserved silages.It is concluded that the need for additives and wilting to ensure satisfactory preservation varies in relation to the variety of grass used and its stage of growth.


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