Low pH Research Articles

Molecular mechanisms and environmental adaptations of flagellar loss and biofilm growth of Rhodanobacter under environmental stress.

Biofilms aid bacterial adhesion to surfaces via direct and indirect mechanisms, and formation of biofilms is considered as an important strategy for adaptation and survival in sub-optimal environmental conditions. However, the molecular underpinnings of biofilm formation in subsurface sediment/groundwater ecosystems where microorganisms often experience fluctuations in nutrient input, pH, nitrate or metal concentrations is underexplored. We examined biofilm formation under different nutrient, pH, metal, and nitrate regimes of 16 Rhodanobacter strains isolated from subsurface groundwater wells spanning diverse pH (3.5 to 5) and nitrate levels (13.7 to 146mM). Eight Rhodanobacter strains demonstrated significant biofilm growth under low pH, suggesting adaptation to survive and grow at low pH. Biofilms intensified under aluminum stress, particularly in strains possessing fewer genetic traits associated with biofilm formation warranting further investigation. Through RB-TnSeq, proteomics, use of specific mutants and transmission electron microscopy analysis, we discovered flagellar loss under aluminum stress, indicating a potential relationship between motility, metal tolerance, and biofilm growth. Comparative genomic analyses revealed absence of flagella and chemotaxis genes, and presence of putative Type VI secretion system in the high biofilm-forming strain FW021-MT20. This study identifies genetic determinants associated with biofilm growth in a predominant environmental genus, Rhodanobacter, under metal stress and identifies traits aiding survival and adaptation to contaminated subsurface environments.

Recent Progress of pH-Responsive Peptides, Polypeptides, and Their Supramolecular Assemblies for Biomedical Applications.

Peptides and polypeptides feature a variety of active functional groups on their side chains (including carboxylic acid, hydroxyl, amino, and thiol groups), enabling diverse chemical modifications. This versatility makes them highly valuable in stimuli-responsive systems. Notably, pH-responsive peptides and polypeptides, due to their ability to respond to pH changes, hold significant promise for applications in cellular pathology and tumor targeting. Extensive researches have highlighted the potentials of low pH insertion peptides (pHLIPs), peptide-drug conjugates (PDCs), and antibody-drug conjugates (ADCs) in biomedicine. Peptide self-assemblies, with their structural stability, ease of regulation, excellent biocompatibility, and biodegradability, offer immense potentials in the development of novel materials and biomedical applications. We also explore specific examples of their applications in drug delivery, tumor targeting, and tissue engineering, while discussing future challenges and potential advancements in the field of pH-responsive self-assembling peptide-based biomaterials.

Biochemical and Structural Characterization of a Novel Psychrophilic Laccase (Multicopper Oxidase) Discovered from Oenococcus oeni 229 (ENOLAB 4002)

Recently, prokaryotic laccases from lactic acid bacteria (LAB), which can degrade biogenic amines, were discovered. A laccase enzyme has been cloned from Oenococcus oeni, a very important LAB in winemaking, and it has been expressed in Escherichia coli. This enzyme has similar characteristics to those previously isolated from LAB as the ability to oxidize canonical substrates such as 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,6-dimethoxyphenol (2,6-DMP), and potassium ferrocyanide K4[Fe(CN6)], and non-conventional substrates as biogenic amines. However, it presents some distinctiveness, the most characteristic being its psychrophilic behaviour, not seen before among these enzymes. Psychrophilic enzymes capable of efficient catalysis at low temperatures are of great interest due to their potential applications in various biotechnological processes. In this study, we report the discovery and characterization of a new psychrophilic laccase, a multicopper oxidase (MCO), from the bacterium Oenococcus oeni. The psychrophilic laccase gene, designated as LcOe 229, was identified through the genomic analysis of O. oeni, a Gram-positive bacterium commonly found in wine fermentation. The gene was successfully cloned and heterologously expressed in Escherichia coli, and the recombinant enzyme was purified to homogeneity. Biochemical characterization of the psychrophilic laccase revealed its optimal activity at low temperatures, with a peak at 10 °C. To our knowledge, this is the lowest optimum temperature described so far for laccases. Furthermore, the psychrophilic laccase demonstrated remarkable stability and activity at low pH (optimum pH 2.5 for ABTS), suggesting its potential for diverse biotechnological applications. The kinetic properties of LcOe 229 were determined, revealing a high catalytic efficiency (kcat/Km) for several substrates at low temperatures. This exceptional cold adaptation of LcOe 229 indicates its potential as a biocatalyst in cold environments or applications requiring low-temperature processes. The crystal structure of the psychrophilic laccase was determined using X-ray crystallography demonstrating structural features similar to other LAB laccases, such as an extended N-terminal and an extended C-terminal end, with the latter containing a disulphide bond. Also, the structure shows two Met residues at the entrance of the T1Cu site, common in LAB laccases, which we suggest could be involved in substrate binding, thus expanding the substrate-binding pocket for laccases. A structural comparison of LcOe 229 with Antarctic laccases has not revealed specific features assigned to cold-active laccases versus mesophilic. Thus, further investigation of this psychrophilic laccase and its engineering could lead to enhanced cold-active enzymes with improved properties for future biotechnological applications. Overall, the discovery of this novel psychrophilic laccase from O. oeni expands our understanding of cold-adapted enzymes and presents new opportunities for their industrial applications in cold environments.

Gold Nanoparticle Mesoporous Carbon Composite as Catalyst for Hydrogen Evolution Reaction

Increased environmental pollution and the shortage of the current fossil fuel energy supply has increased the demand for eco-friendly energy sources. Hydrogen energy has become a potential solution due to its availability and green combustion byproduct. Hydrogen feedstock materials like sodium borohydride (NaBH4) are promising sources of hydrogen; however, the rate at which the hydrogen is released during its reaction with water is slow and requires a stable catalyst. In this study, gold nanoparticles were deposited onto mesoporous carbon to form a nano-composite catalyst (AuNP-MCM), which was then characterized via transmission electron microscopy (TEM), powder X-ray diffraction (P-XRD), and scanning electron microscopy/energy dispersive X-ray spectroscopy (SEM/EDS). The composite’s catalytic ability in a hydrogen evolution reaction was tested under varying conditions, including NaBH4 concentration, pH, and temperature, and it showed an activation of energy of 30.0 kJ mol−1. It was determined that the optimal reaction conditions include high NaBH4 concentrations, lower pH, and higher temperatures. This catalyst, with its stability and competitively low activation energy, makes it a promising material for hydrogen generation.

Impacts of Solidago canadensis and Erigeron canadensis co‐invasion on soil microbial communities

AbstractInvasive plants disrupt the structure and function of ecosystems, causing a decline in their diversity. The coexistence of Solidago canadensis and Erigeron canadensis within shared habitats is a common phenomenon, and the frequent occurrence of co‐invasion has substantial implications for ecosystems. However, the effects of the co‐invasion of two plants on the soil ecosystem across varying degrees of invasion remain uninvestigated. Therefore, we investigated how the co‐invasion of S. canadensis and E. canadensis affects soil physiochemical properties and microorganisms across various degrees of invasion. The invaded regions exhibited significantly lower soil moisture content and pH values than those in the uninvaded areas (p < .05). Additionally, co‐invasion resulted in higher soil organic matter (OM), total nitrogen (TN), total phosphorus (TP), and available phosphorus (AP) contents compared to single invasion by E. canadensis. Furthermore, the invaded regions exhibited a greater diversity of soil bacterial and fungal communities than the uninvaded regions. Under diverse invasion conditions, Actinobacteria and Ascomycota emerged as the predominant phyla of soil bacteria and fungi, respectively, leading to a shift in the soil microbial community structure. Moreover, homogenizing dispersal significantly influenced bacterial community succession, while ecological drift influenced the progression of fungal communities. Using structural equation modeling, this study established significant correlations between soil properties (pH, TN, and nitrate nitrogen) and bacterial interactions (diversity and community assembly) relative to microbial composition. This study revealed the influence of S. canadensis and E. canadensis co‐invasion on soil microbial communities, thereby contributing a theoretical framework delineating the impact of these two invasive plant species.

Morphological and physiological responses of the cosmopolitan marine diatom Thalassiosira rotula to acidification

We investigated the effect of decreased pH on the morphology and nutrient physiology of the cosmopolitan marine diatom Thalassiosira rotula (CCMP3362) by acclimating unialgal cultures to two different CO2 gas concentrations under optimal light, temperature and nutrient conditions. At lower pH (higher CO2), T. rotula exhibited a reduction in cell diameter (7%), surface area (13%), and volume (20%), and an increase in surface area-to-volume ratio (7%). All measures of silicification in T. rotula, i.e., silica (SiO2) quota, rate of silicic acid (Si(OH)4) utilization, and elemental ratios of SiO2:C and SiO2:N, remained unchanged. Similarly, carbon (C) and nitrogen (N) quotas, ratios and utilization rates were mostly unaffected by pH. In contrast, the utilization rate of nitrate (ρNO3) was significantly lower at decreased pH when the rate was normalized by cell number instead of by cell volume. The changes in cell morphology found in this study under low pH were likely not large enough to significantly impact physiological processes and the role of this species in marine food webs and biogeochemical cycles. It is possible that the interactive effects of pH, temperature, light, and nutrient availability characteristic of different regions of the world ocean could result in stronger physiological and morphological responses in this widespread diatom. However, under constant optimal growth conditions, T. rotula was only mildly affected by changes in pH, and in particular the physiology and elemental stoichiometry of C, N and Si were not sensitive to acidification.

Mild alkalinity preheating treatment regulates the heat and ionic strength co-tolerance of whey protein aggregates

A major obstacle to the use of whey protein in protein-enriched sports beverages is the heat-induced gelation of the protein in the presence of salt. In this study, whey protein soluble aggregates (WPSAs) with high tolerance to NaCl and heat were successfully generated by preheating whey protein isolate (WPI) at a low concentration (1 % w/v) and pH 8.5. The suspension of WPSAs (5 % w/v) with 100 mM NaCl maintained clarity, transparency, and good flowability even after 30 min of heating at 100 °C. However, suspensions prepared by untreated WPI turned into milky white gels. WPSAs had a reduced Zeta potential at pH 7 compared to WPI, making them more resistant to the electrostatic screening caused by NaCl. Additionally, WPSAs exhibited reduced sensitivity to heat treatment due to a more compact structure achieved through preheating modification. In light of these findings, a straightforward and effective method was presented for regulating the heat and ionic strength tolerance of whey protein aggregates.

Application of a Novel Dissolution Medium with Lipids for In Vitro Simulation of the Postprandial Gastric Content

Food can change various physiological parameters along the gastrointestinal tract, potentially impacting postprandial drug absorption. It is thus important to consider different in vivo conditions during in vitro studies. Therefore, a novel dissolution medium simulating variable postprandial pH values and lipid concentrations was developed and used in this study. Additionally, by establishing and validating a suitable analytical method, the effects of these parameters on the dissolution of a model drug, cinnarizine, and on its distribution between the lipid and aqueous phases of the medium were studied. Both parameters, pH value and lipid concentration, were shown to influence cinnarizine behavior in the in vitro dissolution studies. The amount of dissolved drug decreased with increasing pH due to cinnarizine’s decreasing solubility. At pH values 5 and 7, the higher concentration of lipids in the medium increased drug dissolution, and most of the dissolved drug was distributed in the lipid phase. In all media with a lower pH of 3, dissolution was fast and complete, with a significant amount of drug distributed in the lipid phase. These results are in accordance with the in vivo observed positive food effect on cinnarizine bioavailability described in the literature. The developed medium, with its ability to easily adjust the pH level and lipid concentration, thus offers a promising tool for assessing the effect of co-ingested food on the dissolution kinetics of poorly soluble drugs.

Soil organic carbon regulation by pH in acidic red soil subjected to long-term liming and straw incorporation

The manipulation of soil pH through liming and straw incorporation plays a pivotal role in influencing soil organic carbon (SOC) dynamics in acidic red soil. This study aimed to assess the impact of these practices on SOC and elucidate the relationship between SOC and pH. Over a 31-year field experiment, seven different fertilization treatments were implemented: unfertilized (CK), nitrogen and potassium fertilizers (NK), NK with lime (NKCa), nitrogen, phosphorous, and potassium fertilizers (NPK), NPK with lime (NPKCa), NPK with straw (NPKS), and NPKS with lime (NPKSCa). Results revealed that liming and straw incorporation significantly elevated soil pH by 0.13–0.73 units. Lime application boosted SOC and mineral-associated organic carbon (MAOC) by 20.2% and 28.7%, respectively, in NK treatment, whereas its impact on SOC in NPK and NPKS treatments were negligible. SOC witnessed a 17.1% increase with NPKS and a 15.2% increase with NPKSCa compared to NPK alone. Notably, NPKS and NPKSCa led to a significant surge in particulate organic carbon (POC) by 19.7% and 37.7%, respectively, albeit NPKSCa reduced MAOC by 14.9% relative to NPK. Linear regression analysis unveiled a positive correlation between POC and soil pH, while SOC and MAOC exhibited an initial rise at lower pH levels followed by stabilization as pH continuously increasing. A partial least squares path model showed two pathways through which pH influenced SOC: firstly, by positively affecting SOC through increasing Fe and Al oxides contents and enhanced aggregate stability, and secondly, by negatively influencing SOC through altered ratios of fungi/bacteria and Gram-positive bacteria/Gram-negative bacteria. In conclusion, the long-term effects of lime and straw application on SOC and MAOC were contingent upon soil pH, with more pronounced positive effects observed at lower pH levels. These findings underscore the importance of considering soil pH when implementing lime and straw strategies to mitigate acidification and regulate SOC in acidic red soil.

Ferrous iron oxidation efficiency and kinetics by indigenous iron-oxidizing bacteria in acid mine drainage

Biological Fe(II) oxidation by iron-oxidising bacteria (FeOBs) at low pH is a cost-effective treatment for acid mine drainage (AMD). However, treatments based on this process are limited because of uncertainties regarding the ability and rate of oxidation of Fe(II) from AMD. In the present study, an indigenous FeOBs consortium was enriched in AMD, and its ability to oxidise Fe(II) is described. The bio-oxidation rate of Fe(II) was 39.1mg/(L·h) under optimal culture conditions [35 ℃, pH 2.0, 500mg/L Fe(II)]. In addition, the oxidation rate equation of Fe(II) could be fitted to a zero-order kinetic model, indicating that Fe(II) was oxidised at a constant rate. Furthermore, a continuous-flow bioreactor was developed to simulate the Fe(II) oxidation efficiency of indigenous FeOBs for in situ AMD biological treatment. The maximum Fe(II) oxidation rate was 22.8mg/(L·h) when the influent Fe(II) load was 32.3mg/(L·h). Acidithiobacillus and Acidiphilium were the dominant species contributing to Fe(II) oxidation in the bioreactor, accounting for 67.7% and 32.8%, respectively. The results will help promote the application of FeOBs in AMD treatment.

Mechanical and physicochemical characteristics of sub-millimeter and macrobubbles: Insights from AFM analysis, zeta potential measurements and rise velocity

This study presents the change in mechanical and physicochemical behaviour of macrobubbles and sub-millimeter bubbles with respect to pH. While macrobubbles conventionally range from 100 μm to 2 mm in diameter, this study focuses on the comparatively less-explored sub-millimeter bubble range, centered around a median diameter of 500 μm. The difference in sub-millimeter bubbles (500 μm) compared to their larger counterparts (1100 μm) within macrobubble size range were evidenced through atomic force microscopy (AFM) analysis, zeta potential measurements, and rise velocity experiments. The adhesion force and energy between bubbles and the silicon nitride cantilever tip showed a decreasing trend with increasing pH, notably diminishing by about 50% beyond pH 9. Although no significant differences were statistically revealed between the two bubble sizes at lower pH, the adhesion force and energy were consistently smaller for macrobubble compared to sub-millimeter bubble at higher pH beyond pH 9. This finding suggests that the electrical double layer (EDL) interactions are influenced by the specific surface area, particularly in the alkaline region. Zeta potential measurements exhibited a cubic trend, with an isoelectric point at pH 3.5 and a minimum zeta potential at pH 11. The study also depicted a similar trend in the rise velocity of bubbles, suggesting a direct relationship between bubble stiffness and its rise velocity. Therefore, changes in pH affected the bubbles' properties, while on the other hand, certain physicochemical properties are also influenced by bubble sizes.

Zn-BTA complex-plugged halloysite nanocontainers for construction of stimuli-responsive epoxy coating

A strategy is proposed to construct stimuli-responsive halloysite nanotubes (HNTs) inhibitor containers in this study. After the benzotriazole (BTA) corrosion inhibitor was loaded into the etched nanotubes (EHNTs), insoluble Zn-BTA complexes as plugs are formed at both ends of the nanotubes by the interaction of BTA with Zn2+ ions (Zn-BTA@EHNTs). The nanocontainers exhibit good pH-sensitivity. At the lowered pH, the extraction of BTA from the nanocontainers is rapid. By contrast, at the neutral pH, the release rate of BTA is relatively slow. Through the electrochemical measurements for the scribed coatings, it is demonstrated that the Zn-BTA@EHNTs-incorporated epoxy coating exhibits distinct stimuli-responsive behavior. And the incorporation of the Zn-BTA@EHNTs remarkably improves the corrosion resistance of the epoxy coating.

Positive Feedback Mechanism in Aristolochic Acid I Exposure-Induced Anemia and DNA Adduct Formation: Implications for Balkan Endemic Nephropathy.

Balkan endemic nephropathy (BEN) is a chronic kidney disease that predominantly affects inhabitants of rural farming communities along the Danube River tributaries in the Balkans. Long-standing research has identified dietary exposure to aristolochic acids (AAs) as the principal toxicological cause. This study investigates the pathophysiological role of anemia in BEN, noting its earlier and more severe manifestation in BEN patients compared to those with other chronic kidney diseases. Utilizing a mouse model, our research demonstrates that prolonged exposure to aristolochic acid I (AA-I) (the most prevalent AA variant) leads to significant red blood cell depletion through DNA damage, such as DNA adduct formation in bone marrow, prior to observable kidney function decline. Furthermore, in vitro experiments with kidney cells exposed to lowered oxygen and pH conditions mimicking an anemia environment show enhanced DNA adduct formation, suggesting increased AA-I mutagenicity and carcinogenicity. These findings indicate for the first time a positive feedback mechanism of AA-induced anemia, DNA damage, and kidney impairment in BEN progression. These results not only advance our understanding of the underlying mechanisms of BEN but also highlight anemia as a potential target for early BEN diagnosis and therapy.

Understanding zinc isotopic signatures in volcanic lakes

Magmatic activity can be monitored through the chemical variations of volcanic lakes. While those lakes can be influenced by direct magmatic input, water-rock interaction processes in hydrothermal system also have a large influence. Disentangling those effects is critical to use crater lakes for volcanic activity monitoring purposes. Zinc is a volatile metal that is easily degassed and can be incorporated into fluid circulation. During degassing, Zn isotopes can be fractionated and can thus be a potential tracer of magmatic input. Here we present a novel approach using stable Zn isotopic systems to understand how dissolution/precipitation processes affect isotopic compositions of volcanic lakes and if they can be used as tracers of magmatic activity. We investigate Zn isotopes expressed as δ66Zn for four volcanic lakes ranging from hyperacidic to neutral waters. Two of them, Kawah Ijen (Indonesia) and Santa Ana (El Salvador) volcanic lakes have been sampled from 2010 to 2012 and 2000–2002, respectively, and show a clear magmatic input with extremely low pH [0.1–1]. Taal (Philippines, pH ≈ 3) and Kelud (Indonesia, pH ≈ 6.5) volcanic lakes were sampled from 2017 to 2019 and 2003–2007 respectively, and are dominated by hydrothermal rather than magmatic inputs. Hyper-acidic lakes show minimal isotopic variation over the investigated time ranges, with δ66Zn values similar to their basalt-andesitic host rocks (0.26 ± 0.08 ‰). In contrast, we observe large δ66Zn fractionation for the Taal waters (0.4 ± 0.01 ‰) compared to their basalt-andesitic host rocks but no δ66Zn temporal variations. Kelud waters show significant δ66Zn (−0.08 ± 0.06 ‰ to 0.43 ± 0.06 ‰) temporal variation. This could be related to an evolution of the lake water chemistry as the sampling period occurred between two increases in lake temperature culminating into a lava dome eruption in 2007. For Taal and Kelud volcanic lakes, enrichment factors (EF) and PHREEQc modelling indicate that Zn is depleted and suggest that Zn could precipitate in the hydrothermal system, hence fractionating Zn isotopes during these precipitation processes. This contrasts to Zn elemental and isotope values from Kawah Ijen and Santa Ana with an EF ≈ 1 suggesting that Zn is derived from host rock dissolution, in agreement with their δ66Zn. These data are still limited and a longer observation is needed in the future to highlight significant geochemical changes in volcanic lakes and their relation to volcanic activity.

AIE-active large Stokes-shift BODIPY Functionalized with Carbazolyl for Lysosome-Targeted Imaging in Living Cells

A large number of studies have shown that lysosomal microcircumstances changes can affect many physiological and pathological processes at the cellular level. However, the visual detection of lysosomal microcircumstances is relatively difficult due to low pH (4.5–6.0) value in lysosomal that require the probe not only stable under acidic condition but also has a good localization effect to lysosomal. Obviously, novel fluorescent which possessed both acidic stability and lysosomal-target property together with lysosomal viscosity active is highly demanded. Herein, a novel BODIPY molecular CarBDP based on carbazole group was rationally designed and synthesized for the lysosomal imaging. CarBDP exhibited AIE feature with a large Stokes shift of up to 157 nm. More importantly, co-localization assay of the CarBDP-treated MCF-7 cells indicated that CarBDP has a good localization effect on lysosomal (Rr = 0.7109) due to the carbazole group while the normal BODIPY that without carbazole group (PhBDP) shows poor localization performance, this was the first time that a small molecule can locate lysosomes only based on carbazole group. CarBDP exhibits strong solid emission with long fluorescence decay lifetime (τ = 44.54 ns) and was stable under acid condition.The probe CarBDP assembled with carbazole group was successfully utilized for lysosomal localization and mapping lysosomal viscosity in live cells, which provides a novel candidate tool for the determination of lysosomal microcircumstances.

Enhanced bioleaching of spent Li-ion batteries using A. ferrooxidans by application of external magnetic field

Recycling spent batteries is increasingly important for the sustainable use of Li-ion batteries (LIBs) and for countering the supply uncertainty of critical raw minerals (Li, Co, and Ni). Bioleaching, which uses microorganisms to extract valuable metals, is both economical and environmentally safe compared to other recycling methods, but its practical application is impaired by slow kinetics. Accelerating the process is a key for bioleaching spent LIBs on an industrial scale. Acidithiobacillus ferrooxidans (A. ferrooxidans), which thrives in extremely low pH conditions, has long been explored for bioleaching of spent LIBs. Metabolism of A. ferrooxidans involves the oxidation of magnetic Fe2+ and produces intracellular magnetic nanoparticles. The possibility of accelerating the leaching kinetics of A. ferrooxidans by the application of an external magnetic field is explored in this work. A weak static magnetic field is applied during the bioleaching of spent LIBs to recover Li, Ni, and Co using A. ferrooxidans. It is determined that 3 mT is the optimal field strength which allows the leaching efficiency of Li to reach 100% after only 2 days of leaching at a pulp density of 3 w/v % while without the external magnetic field, the leaching efficiency is limited to 57% even after 4 days. The leaching efficiency of Ni and Co also increases by nearly three-fold to >80% after 4 days of leaching. The proposed magnetic field-assisted bioleaching of spent LIBs using A. ferrooxidans substantially improves the leaching kinetics and thus the cost-effectiveness of the bioleaching process with minimal environmental impact, hence enabling environment-friendly recycling of raw materials that are increasingly becoming scarce. The positive effect of an external magnetic field on the metabolism of A. ferrooxidans demonstrated in this work provide a new set of tools to engineer the bioleaching process and the possibility for genetic modification of acidophile bacteria, especially targeted for magnetic enhancement.

Development of a scalable, high-anthocyanin and low-acidity natural red food colorant from Hibiscus sabdariffa L.

The hibiscus calyx contains 0.3–2.4% total anthocyanins, and is a promising source for naturally red food colorants. In this study, commercially available hibiscus calyces were subjected to ethanolic-aqueous extraction and chromatographic enrichment with the XAD-7HP resin, to create scalable, high-anthocyanin and low-acidity natural food colorants. Anthocyanins, organic and phenolic acids were monitored after each step using UHPLC-DAD and UHPLC-QQQ/MS. 75.67% total anthocyanins were recovered from calyces after double extractions, and the content increased by 8.50–14.90 times after the column enrichment, reaching 14.51–31.90% (by dry weight) in the final product. Chromatographic fractionation was also shown to effectively increase the total phenolic acids by 11.01–16.22 times, and remove an average of 98.58% of the total organic acids. High intensity redness at pH 2.5–3.5 indicated that the final product may be a promising, versatile natural food and beverage colorant in low pH products.

Application of acid-activated near-infrared viscosity fluorescent probe targeting lysosomes in cancer visualization

The higher viscosity and lower pH in lysosomes of cancer cells highlight their potential as biomarkers for cancer. Therefore, the development of acid-activated viscosity fluorescent probes is significant for the early diagnosis and treatment of cancer. Based on this, we have designed and synthesized a near-infrared fluorescent probe based on the 2-(2-hydroxyphenyl)benzothiazole (HBT) group, namely HBTH, to monitor the viscosity changes within lysosomes. It has been demonstrated that HBTH was extremely sensitive to viscosity, with a strong linear relationship between fluorescence intensity and log(viscosity) within the range of (logη) = 0–3.06 (a correlation coefficient of 0.98), proving its capability for quantitative viscosity measurement. In particular, the most obvious fluorescence enhancement of HBTH was only efficiently triggered by the combined effect of low pH and high viscosity. Furthermore, HBTH can rapidly localize to lysosomes by wash-free procedure at a low concentration (100 nM) and achieve high-fidelity imaging within 20 s. It can also monitor the dynamic processes of lysosomes in cells, viscosity changes under drug stimuli, and lysosomal behavior during mitophagy. Importantly, HBTH is capable of identifying tumors in tumor-bearing nude mice through in vivo imaging. These features make HBTH a powerful tool for the early diagnosis and treatment of cancer.

Rice-crayfish integrated system enhances global warming potential via increasing methane emission mainly driven by continuous deep flooding

The conversion from rice monoculture (RM) to rice-crayfish integrated system (RCIS) could change soil physicochemical properties and microorganisms related to greenhouse gas (GHG) emissions. Nevertheless, it is still unclear the responses of GHG emissions and global warming potential (GWP) from paddy fields to RCIS. Here, we conducted a field experiment to investigate the changes in the emissions of methane (CH4) and nitrous oxide (N2O), GWP, soil physicochemical properties and the associated microbial abundances (methanogen, methanotrophs, denitrifier and nitrifier) under RCIS in Gaoyou City, Jiangsu Province, China from 2022 to 2023. There were three treatments, including RM, rice monoculture with continuous deep flooding (RMF) and RCIS. Compared with RM, RCIS and RMF all significantly increased CH4 emission and decreased N2O emission. But the effectiveness of increasing CH4 emission and decreasing N2O emission was larger under RCIS than RMF. The increased CH4 emission was due to the significantly higher mcrA gene abundance and ratio of mcrA/pmoA under RCIS. While the significantly higher nosZ gene abundance and ratio of nosZ/(amoA+nirK+nirS) under RCIS were responsible for its reduced N2O emission. Furthermore, the increases in gene abundances of mcrA and nosZ under RCIS were closely correlated with the significantly lower redox potential and pH as well as the significantly higher contents of dissolved organic carbon, ammonium and nitrate in soil. Averaged across two years, GWP under RCIS and RMF were 4.6-fold and 3.4-fold that under RM, respectively. This revealed that continuous deep flooding had a greater effectiveness in increasing GWP than crayfish culture under RCIS. Notably, the contribution rates of CH4 emission to the total GWP decreased in the order of RCIS (98.6%) > RMF (97.5%) > RM (86.2%). Besides, greenhouse gas intensity under RCIS was 5.1-fold that of RM due to its enhanced GWP and reduced rice grain yield. In summary, RCIS could enhance GWP from paddy fields through increasing CH4 emission mainly caused by the continuous deep flooding.

Tad pili contribute to the virulence and biofilm formation of virulent Aeromonas hydrophila

Type IV pili (T4P) are versatile proteinaceous protrusions that mediate diverse bacterial processes, including adhesion, motility, and biofilm formation. Aeromonas hydrophila, a Gram-negative facultative anaerobe, causes disease in a wide range of hosts. Previously, we reported the presence of a unique Type IV class C pilus, known as tight adherence (Tad), in virulent Aeromonas hydrophila (vAh). In the present study, we sought to functionalize the role of Tad pili in the pathogenicity of A. hydrophila ML09-119. Through a comprehensive comparative genomics analysis of 170 A. hydrophila genomes, the conserved presence of the Tad operon in vAh isolates was confirmed, suggesting its potential contribution to pathogenicity. Herein, the entire Tad operon was knocked out from A. hydrophila ML09-119 to elucidate its specific role in A. hydrophila virulence. The absence of the Tad operon did not affect growth kinetics but significantly reduced virulence in catfish fingerlings, highlighting the essential role of the Tad operon during infection. Biofilm formation of A. hydrophila ML09-119 was significantly decreased in the Tad operon deletant. Absence of the Tad operon had no effect on sensitivity to other environmental stressors, including hydrogen peroxide, osmolarity, alkalinity, and temperature; however, it was more sensitive to low pH conditions. Scanning electron microscopy revealed that the Tad mutant had a rougher surface structure during log phase growth than the wildtype strain, indicating the absence of Tad impacts the outer surface of vAh during cell division, of which the biological consequences are unknown. These findings highlight the role of Tad in vAh pathogenesis and biofilm formation, signifying the importance of T4P in bacterial infections.