Cystic fibrosis (CF) is a life-shortening disease caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene. Although bacterial lung infection and the resulting inflammation cause most of the morbidity and mortality, how the loss of CFTR function first disrupts airway host defence has remained uncertain. To investigate the abnormalities that impair elimination when a bacterium lands on the pristine surface of a newborn CF airway, we interrogated the viability of individual bacteria immobilized on solid grids and placed onto the airway surface. As a model, we studied CF pigs, which spontaneously develop hallmark features of CF lung disease. At birth, their lungs lack infection and inflammation, but have a reduced ability to eradicate bacteria. Here we show that in newborn wild-type pigs, the thin layer of airway surface liquid (ASL) rapidly kills bacteria in vivo, when removed from the lung and in primary epithelial cultures. Lack of CFTR reduces bacterial killing. We found that the ASL pH was more acidic in CF pigs, and reducing pH inhibited the antimicrobial activity of ASL. Reducing ASL pH diminished bacterial killing in wild-type pigs, and, conversely, increasing ASL pH rescued killing in CF pigs. These results directly link the initial host defence defect to the loss of CFTR, an anion channel that facilitates HCO3− transport. Without CFTR, airway epithelial HCO3− secretion is defective, the ASL pH falls and inhibits antimicrobial function, and thereby impairs the killing of bacteria that enter the newborn lung. These findings suggest that increasing ASL pH might prevent the initial infection in patients with CF, and that assaying bacterial killing could report on the benefit of therapeutic interventions.
Aqueous solutions of lead salts (1, 2) and saturated solutions of lead hydroxide (1) have been used as stains to enhance the electron-scattering properties of components of biological materials examined in the electron microscope. Saturated solutions of lead hydroxide (1), while staining more intensely than either lead acetate or monobasic lead acetate (l , 2), form insoluble lead carbonate upon exposure to air. The avoidance of such precipitates which contaminate surfaces of sections during staining has been the stimulus for the development of elaborate procedures for exclusion of air or carbon dioxide (3, 4). Several modifications of Watson's lead hydroxide stain (1) have recently appeared (5-7). All utilize relatively high pH (approximately 12) and one contains small amounts of tartrate (6), a relatively weak complexing agent (8), in addition to lead. These modified lead stains are less liable to contaminate the surface of the section with precipitated stain products. The stain reported here differs from previous alkaline lead stains in that the chelating agent, citrate, is in sufficient excess to sequester all lead present. Lead citrate, soluble in high concentrations in basic solutions, is a chelate compound with an apparent association constant (log Ka) between ligand and lead ion of 6.5 (9). Tissue binding sites, presumably organophosphates, and other anionic species present in biological components following fixation, dehydration, and plastic embedding apparently have a greater affinity for this cation than lead citrate inasmuch as cellular and extracellular structures in the section sequester lead from the staining solution. Alkaline lead citrate solutions are less likely to contaminate sections, as no precipitates form when droplets of fresh staining solution are exposed to air for periods of up to 30 minutes. The resultant staining of the sections is of high intensity in sections of Aralditeor Epon-embedded material. Cytoplasmic membranes, ribosomes, glycogen, and nuclear material are stained (Figs. 1 to 3). STAIN SOLUTION: Lead citrate is prepared by
Summary We demonstrate a long-lifetime, aqueous redox-flow battery that can operate at a pH as low as 12 while maintaining an open-circuit voltage of over 1 V. We functionalized 2,6-dihydroxyanthraquinone (2,6-DHAQ) with highly alkali-soluble carboxylate terminal groups. The resulting negative electrolyte material 4,4′-((9,10-anthraquinone-2,6-diyl)dioxy)dibutyrate (2,6-DBEAQ) was six times more soluble than 2,6-DHAQ at pH 12. Symmetric cell cycling with 2,6-DBEAQ on both sides of the cell demonstrates a capacity fade rate of
The cytoplasmic membrane of a prokaryotic cell consists of a lipid bilayer or a monolayer that shields the cellular content from the environment. In addition, the membrane contains proteins that are responsible for transport of proteins and metabolites as well as for signalling and energy transduction. Maintenance of the functionality of the membrane during changing environmental conditions relies on the cell’s potential to rapidly adjust the lipid composition of its membrane. Despite the fundamental chemical differences between bacterial ester lipids and archaeal ether lipids, both types are functional under a wide range of environmental conditions. We here provide an overview of archaeal and bacterial strategies of changing the lipid compositions of their membranes. Some molecular adjustments are unique for archaea or bacteria, whereas others are shared between the two domains. Strikingly, shared adjustments were predominantly observed near the growth boundaries of bacteria. Here, we demonstrate that the presence of membrane spanning ether-lipids and methyl branches shows a striking relationship with the growth boundaries of archaea and bacteria.
Significance The climate and ocean pH of the early Earth are important for understanding the origin and early evolution of life. However, estimates of early climate range from below freezing to over 70 °C, and ocean pH estimates span from strongly acidic to alkaline. To better constrain environmental conditions, we applied a self-consistent geological carbon cycle model to the last 4 billion years. The model predicts a temperate (0–50 °C) climate and circumneutral ocean pH throughout the Precambrian due to stabilizing feedbacks from continental and seafloor weathering. These environmental conditions under which life emerged and diversified were akin to the modern Earth. Similar stabilizing feedbacks on climate and ocean pH may operate on earthlike exoplanets, implying life elsewhere could emerge in comparable environments. The early Earth’s environment is controversial. Climatic estimates range from hot to glacial, and inferred marine pH spans strongly alkaline to acidic. Better understanding of early climate and ocean chemistry would improve our knowledge of the origin of life and its coevolution with the environment. Here, we use a geological carbon cycle model with ocean chemistry to calculate self-consistent histories of climate and ocean pH. Our carbon cycle model includes an empirically justified temperature and pH dependence of seafloor weathering, allowing the relative importance of continental and seafloor weathering to be evaluated. We find that the Archean climate was likely temperate (0–50 °C) due to the combined negative feedbacks of continental and seafloor weathering. Ocean pH evolves monotonically from 6.6−0.4+0.6 (2σ) at 4.0 Ga to 7.0−0.5+0.7 (2σ) at the Archean–Proterozoic boundary, and to 7.9−0.2+0.1 (2σ) at the Proterozoic–Phanerozoic boundary. This evolution is driven by the secular decline of pCO2, which in turn is a consequence of increasing solar luminosity, but is moderated by carbonate alkalinity delivered from continental and seafloor weathering. Archean seafloor weathering may have been a comparable carbon sink to continental weathering, but is less dominant than previously assumed, and would not have induced global glaciation. We show how these conclusions are robust to a wide range of scenarios for continental growth, internal heat flow evolution and outgassing history, greenhouse gas abundances, and changes in the biotic enhancement of weathering.
pH influences the occurrence and distribution of microorganisms. Microbes typically live over a range of 3 to 4 pH units and are described as acidophiles, neutrophiles, and alkaliphiles, depending on the optimal pH for growth. Their growth rates vary with pH along bell- or triangle-shaped curve, which reflects pH limits of cell structure integrity and the interference of pH with cell metabolism. We propose that pH can also affect the thermodynamics and kinetics of microbial respiration, which then help shape the composition and function of microbial communities. Here we use geochemical reaction modeling to examine how environmental pH controls the energy yields of common redox reactions in anoxic environments, including syntrophic oxidation, iron reduction, sulfate reduction, and methanogenesis. The results reveal that environmental pH changes the energy yields both directly and indirectly. The direct change applies to the reactions that consume or produce protons whereas the indirect effect, which applies to all redox reactions, comes from the regulation of chemical speciation by pH. The results also show that the energy yields respond strongly to pH variation, which may modulate microbial interactions and help give rise to the pH limits of microbial metabolisms. These results underscore the importance of pH as a control on microbial metabolisms and provide insight into potential impacts of pH variation on the composition and activity of microbial communities. In a companion paper, we continue to explore how the kinetics of microbial metabolisms responds to pH variations, and how these responses control the outcome of microbial interactions, including the activity and membership of microbial consortia.
The pH of the tumor microenvironment drives the metastatic phenotype and chemotherapeutic resistance of tumors. Understanding the mechanisms underlying this pH-dependent phenomenon will lead to improved drug delivery and allow the identification of new therapeutic targets. This includes an understanding of the role pH plays in primary tumor cells, and the regulatory factors that permit cancer cells to thrive. Over the last decade, carbonic anhydrases (CAs) have been shown to be important mediators of tumor cell pH by modulating the bicarbonate and proton concentrations for cell survival and proliferation. This has prompted an effort to inhibit specific CA isoforms, as an anti-cancer therapeutic strategy. Of the 12 active CA isoforms, two, CA IX and XII, have been considered anti-cancer targets. However, other CA isoforms also show similar activity and tissue distribution in cancers and have not been considered as therapeutic targets for cancer treatment. In this review, we consider all the CA isoforms and their possible role in tumors and their potential as targets for cancer therapy.
Biochars are potentially effective sorbents for NH4+ and NO3− in water treatment and soil applications. Here we compare NH4+ and NO3− sorption rates to acid-washed biochars produced from red oak (Quercus rubra) and corn stover (Zea mays) at three pyrolysis temperatures (400, 500 and 600 °C) and a range of solution pHs (3.5–7.5). Additionally, we examined sorption mechanisms by quantification of NH4+ and NO3− sorption, as well as Ca2+ and Cl− displacement for corn stover biochars. Solution pH curves showed that NH4+ sorption was maximized (0.7–0.8 mg N g−1) with low pyrolysis temperature (400 °C) biochar at near neutral pH (7.0–7.5), whereas NO3− sorption was maximized (1.4–1.5 mg N g−1) with high pyrolysis temperatures (600 °C) and low pH (3.5–4). The Langmuir (r2 = 0.90–1.00) and Freundlich (r2 = 0.81–0.97) models were good predictors for both NH4+ (pH 7) and NO3− (pH 3.7) sorption isotherms. Lastly, NH4+ and NO3− displaced Ca2+ and Cl−, respectively, from previously CaCl2-saturated corn stover biochars. Results from the pH curves, Langmuir isotherms, and cation displacement curves all support the predominance of ion exchange mechanisms. Our results demonstrate the importance of solution pH and chemical composition in influencing NH4+ and NO3− sorption capacities of biochar.
Regarded as a silent epidemic, chronic wounds are a global public health issue. Wound healing is a complex, synchronized cascade of physiological processes restoring the anatomic and functional integrity of the skin; however, chronic wounds fail to proceed through the wound healing cascade. Wound pH oscillates during wound healing, usually traversing from a neutral pH to an acidic pH, while chronic wounds perpetuate in an elevated alkaline milieu. Although a neglected clinical parameter, pH has implications for relatively all pathologies of wound healing affecting oxygen release, angiogenesis, protease activity, bacterial toxicity and antimicrobial activity. Despite the array of wound healing products currently marketed, understanding the implications of pH on arresting wound healing can stimulate innovation within this vast market.
Abstract Soil exoenzymes released by microorganisms break down organic matter and are crucial in regulating C, N and P cycling. Soil pH is known to influence enzyme activity, and is also a strong driver of microbial community composition; but little is known about how alterations in soil pH affect enzymatic activity and how this is mediated by microbial communities. To assess long term enzymatic adaptation to soil pH, we conducted enzyme assays at buffered pH levels on two historically managed soils maintained at either pH 5 or 7 from the Rothamsted Park Grass Long-term experiment. The pH optima for a range of exoenzymes involved in C, N, P cycling, differed between the two soils, the direction of the shift being toward the source soil pH, indicating the production of pH adapted isoenzymes by the soil microbial community. Soil bacterial and fungal communities determined by amplicon sequencing were clearly distinct between pH 5 and soil pH 7 soils, possibly explaining differences in enzymatic responses. Furthermore, β-glucosidase gene sequences extracted from metagenomes revealed an increased abundance of Acidobacterial producers in the pH 5 soils, and Actinobacteria in pH 7 soils. Our findings demonstrate that the pH optimum of soil exoenzymes adapt to long term changes in soil pH, the direction being dependent on the soil pH shift; and we provide further evidence that changes in functional microbial communities may underpin this phenomena, though new research is now needed to directly link change in enzyme activity optima with microbial communities. More generally, our new findings have large implications for modelling the efficiency of different microbial enzymatic processes under changing environmental conditions.
Distinct subcellular pH levels, especially in lysosomes and endosomes, are essential for the degradation, modification, sorting, accumulation, and secretion of macromolecules. Here, we engineered a novel genetically encoded pH probe by fusing the pH-stable cyan fluorescent protein (FP) variant, mTurquoise2, to the highly pH-sensitive enhanced yellow fluorescent protein, EYFP. This approach yielded a ratiometric biosensor—referred to as pH-Lemon—optimized for live imaging of distinct pH conditions within acidic cellular compartments. Protonation of pH-Lemon under acidic conditions significantly decreases the yellow fluorescence while the cyan fluorescence increases due to reduced Förster resonance energy transfer (FRET) efficiency. Because of its freely reversible and ratiometric responses, pH-Lemon represents a fluorescent biosensor for pH dynamics. pH-Lemon also shows a sizable pH-dependent fluorescence lifetime change that can be used in fluorescence lifetime imaging microscopy as an alternative observation method for the study of pH in acidic cellular compartments. Fusion of pH-Lemon to the protein microtubule-associated protein 1A/1B-light chain 3B (LC3B), a specific marker of autophagic membranes, resulted in its targeting within autolysosomes of HeLa cells. Moreover, fusion of pH-Lemon to a glycophosphatidylinositol (GPI) anchor allowed us to monitor the entire luminal space of the secretory pathway and the exoplasmic leaflet of the plasma membrane. Utilizing this new pH probe, we revealed neutral and acidic vesicles and substructures inside cells, highlighting compartments of distinct pH throughout the endomembrane system. These data demonstrate, that this novel pH sensor, pH-Lemon, is very suitable for the study of local pH dynamics of subcellular microstructures in living cells.