Understanding phosphorus mobilization mechanisms in acidic soil amended with calcium-silicon-magnesium-potassium fertilizer.

Calcium-silicon-magnesium-potassium fertilizer (CSMP) is usually used as an amendment to counteract soil acidification caused by historical excessive nitrogen (N) applications. However, the impact of CSMP addition on phosphorus (P) mobilization in acidic soils and the related mechanisms are not fully understood. Specifically, a knowledge gap exists with regards to changes in soil extracellular enzymes that contribute to P release. Such a knowledge gap was investigated by an incubation study with four treatments: i) initial soil (Control), ii) urea (60 mg kg-1) addition (U); iii) CSMP (1%) addition (CSMP) and iv) urea (60 mg kg-1) and CSMP (1%) additions (U + CSMP). Phosphorus mobilization induced by different processes was distinguished by biologically based P extraction. The Langmuir equation, K edge X-ray absorption near-edge structure spectroscopy, and ecoenzyme vector analysis according to the extracellular enzyme activity stoichiometry were deployed to investigate soil P sorption intensity, precipitation species, and microbial-driven turnover of organophosphorus. Results showed that CaCl2 extractable P (or citric acid extractable P) content increased by 63.4% (or 39.2%) in the soil with CSMP addition, compared with the study control. The accelerated mobilization of aluminum (Al)/iron (Fe)-bound P after CSMP addition, indicated by the reduction of the sum of FePO4·2H2O and AlPO4 proportion, contributed to this increase. The decrease of P sorption capacity can also be responsible for it. The CSMP addition increased enzyme extractable P in the soil nearly 7-fold and mitigated the limitations of carbon (C) and P for soil microorganisms (indicated by the enzyme stoichiometry and ecoenzyme vector analysis), suggesting that microbial turnover processes also contribute to P mobilization in amended acidic soil. These findings indicate that the P mobilization in CSMP amended acidic soil not only attributed to both decreasing P sorption capacity and dissolving phosphate precipitation, but also to the increase of the microbial turnover of the organophosphorus pool.

Medial Septal Glutamatergic Neurons Modulate States of Consciousness during Sevoflurane Anesthesia in Mice.

BACKGROUND: Multiple neural structures involved in maintaining wakefulness have been found to promote arousal from general anesthesia. The medial septum is a critical region that modulates arousal behavior. This study hypothesized that glutamatergic neurons in the medial septum play a crucial role in regulating states of consciousness during sevoflurane general anesthesia. METHODS: Adult male mice were used in this study. The effects of sevoflurane anesthesia on neuronal activity were determined by fiber photometry. Lesions and chemogenetic manipulations were used to study the effects of the altered activity of medial septal glutamatergic neurons on anesthesia induction, emergence, and sensitivity to sevoflurane. Optogenetic stimulation was used to observe the role of acute activation of medial septal glutamatergic neurons on cortical activity and behavioral changes during sevoflurane-induced continuous steady state of general anesthesia and burst suppression state. RESULTS: The authors found that medial septal glutamatergic neuronal activity decreased during sevoflurane anesthesia induction and recovered in the early period of emergence. Chemogenetic activation of medial septal glutamatergic neurons prolonged the induction time (mean ± SD, hM3Dq-clozapine N-oxide vs. hM3Dq-saline, 297.5 ± 60.1 s vs. 229.4 ± 29.9 s, P < 0.001, n = 11) and decreased the emergence time (53.2 ± 11.8 s vs. 77.5 ± 33.5 s, P = 0.025, n = 11). Lesions or chemogenetic inhibition of these neurons produced the opposite effects. During steady state of general anesthesia and deep anesthesia-induced burst suppression state, acute optogenetic activation of medial septal glutamatergic neurons induced cortical activation and behavioral emergence. CONCLUSIONS: The study findings reveal that activation of medial septal glutamatergic neurons has arousal-promoting effects during sevoflurane anesthesia in male mice. The activation of these neurons prolongs the induction and accelerates the emergence of anesthesia.

Medial septum glutamatergic neurons modulate nociception in chronic neuropathic pain via projections to lateral hypothalamus.

The medial septum (MS) contributes in pain processing and regulation, especially concerning persistent nociception. However, the role of MS glutamatergic neurons in pain and the underlying neural circuit mechanisms in pain remain poorly understood. In this study, chronic constrictive injury of the sciatic nerve (CCI) surgery was performed to induce thermal and mechanical hyperalgesia in mice. The chemogenetic activation of MS glutamatergic neurons decreased pain thresholds in naïve mice. In contrast, inhibition or ablation of these neurons has improved nociception thresholds in naïve mice and relieved thermal and mechanical hyperalgesia in CCI mice. Anterograde viral tracing revealed that MS glutamatergic neurons had projections to the lateral hypothalamus (LH) and supramammillary nucleus (SuM). We further demonstrated that MS glutamatergic neurons regulate pain thresholds by projecting to LH but not SuM, because the inhibition of MS-LH glutamatergic projections suppressed pain thresholds in CCI and naïve mice, yet, optogenetic activation or inhibition of MS-SuM glutamatergic projections had no effect on pain thresholds in naïve mice. In conclusion, our results reveal that MS glutamatergic neurons play a significant role in regulating pain perception and decipher that MS glutamatergic neurons modulate nociception via projections to LH.

Fate of 15N-labelled urea as affected by long-term manure substitution.

Quantifying the fate of fertilizer nitrogen (N) is essential to develop more sustainable agricultural fertilization practices. However, the fate of chemical fertilizer N, particularly in long-term manure substitution treatment regimes, is not fully understood. The present study aimed to investigate the fate of 15N-labelled urea in a chemical fertilizer treatment (CF, 240 kg 15N ha-1) and N manure 50 % substitution treatment (1/2N + M, 120 kg 15N ha-1 + 120 kg manure N ha-1) in two continuous crop seasons, based on a 10-year long-term experiment in the North China Plain (NCP). The results showed that manure substitution greatly enhanced 15N use efficiency (15NUE) (39.9 % vs. 31.3 %) and suppressed 15N loss (6.9 % vs. 7.5 %) compared with the CF treatment in the first crop. However, the N2O emissions factor in the 1/2N + M treatment was increased by 0.1 % (0.5 kg 15N ha-1 for CF vs. 0.4 kg 15N ha-1 for 1/2N + M) compared with the CF treatment, although N leaching and NH3 volatilization rates decreased by 0.2 % (10.8 kg 15N ha-1 for CF vs. 5.1 kg 15N ha-1 for 1/2N + M) and 0.5 % (6.6 kg 15N ha-1 for CF vs. 2.8 kg 15N ha-1 for 1/2N + M), respectively. In which, only NH3 volatilization presented significantly difference between treatments. It is important to note that in the second crop, the residual 15N in soil (0-20 cm) remained mostly in the soil for the CF (79.1 %) and the 1/2N + M treatment (85.3 %), and contributed less to crop N uptake (3.3 % vs. 0.8 %) and leached losses (2.2 % vs. 0.6 %). This proved that manure substitution could enhance the stabilization of chemical N. These results suggested that long-term manure substitution effectively increases NUE, suppresses N loss, and improves N stabilization in soil, but negative impacts such as N2O emissions due to climate change should be investigated further.

Enhanced phosphorus fixation in red mud-amended acidic soil subjected to periodic flooding-drying and straw incorporation.

Globally, red mud is a solid waste from the aluminum industry, which is rich in iron oxides. It is an effective soil amendment in agriculture that protects connected waters from legacy diffuse phosphorus (P) soil losses. However, other management practices such as flooding and drying and/or organic carbon inputs could potentially alter P fixation in these red mud-amended soils thereby releasing P to waters. The present study was designed and conducted to monitor the mobilization of P in a red mud-amended acidic soil subjected to periodic flooding-drying, straw incorporation, and a mix of both management practices. Sequential extraction and K edge X-ray absorption near-edge structure spectroscopy (k-XANES) were employed to distinguish P fractions/species and the Langmuir model was fitted to evaluate soil P sorption capacity. The content of labile P indicated by CaCl2-P was increased significantly by 101% and 28.7% in the straw incorporation and periodic flooding-drying treatments, while it decreased significantly by 22.3% in the combined periodic flooding-drying with straw incorporation treatment, compared with Control. The inherent phosphate contained in sorghum straw, and the enhanced iron (Fe) reduction and dissolution of Calcium (Ca)-bound P induced by straw addition contributed to mobilization of P in the straw incorporation treatment. In contrast, the increased poorly crystalline Al/Fe oxides-bound P and occluded Fe-bound P fraction in the combined periodic flooding-drying with straw incorporation treatment explains the decrease in CaCl2-P. Furthermore, the increased soil P sorption capacity and the decreased P desorption rate were also responsible for the reduced P loss risk in the treatment. The results of structural equation modelling (SEM) indicated that organically complexed Fe and Fe-bound P were directly affecting P mobilization in the amended soil. Overall, the present study shows that appropriate flooding-drying events coupled with straw incorporation could be a mitigation practice for stabilizing P in red mud-amended soil. However, before it can be applied on a wide scale, multi-point and field trials should be carried out to further evaluate actual environmental implications.

Neural and molecular investigation into the paraventricular thalamic-nucleus accumbens circuit for pain sensation and non-opioid analgesia.

The paucity of medications with novel mechanisms for pain treatment combined with the severe adverse effects of opioid analgesics has led to an imperative pursuit of non-opioid analgesia and a better understanding of pain mechanisms. Here, we identify the putative glutamatergic inputs from the paraventricular thalamic nucleus to the nucleus accumbens (PVTGlut→NAc) as a novel neural circuit for pain sensation and non-opioid analgesia. Our in vivo fiber photometry and in vitro electrophysiology experiments found that PVTGlut→NAc neuronal activity increased in response to acute thermal/mechanical stimuli and persistent inflammatory pain. Direct optogenetic activation of these neurons in the PVT or their terminals in the NAc induced pain-like behaviors. Conversely, inhibition of PVTGlut→NAc neurons or their NAc terminals exhibited a potent analgesic effect in both naïve and pathological pain mice, which could not be prevented by pretreatment of naloxone, an opioid receptor antagonist. Anterograde trans-synaptic optogenetic experiments consistently demonstrated that the PVTGlut→NAc circuit bi-directionally modulates pain behaviors. Furthermore, circuit-specific molecular profiling and pharmacological studies revealed dopamine receptor 3 as a candidate target for pain modulation and non-opioid analgesic development. Taken together, these findings provide a previously unknown neural circuit for pain sensation and non-opioid analgesia and a valuable molecular target for developing future safer medication.

Alteration of neural activity and neuroinflammatory factors in the insular cortex of mice with corneal neuropathic pain.

Dry eye disease (DED) affects nearly 55% of people worldwide; several studies have proposed that central sensitization and neuroinflammation may contribute to the developing corneal neuropathic pain of DED, while the underlying mechanisms of this contribution remain to be investigated. Excision of extra orbital lacrimal glands established the dry eye model. Corneal hypersensitivity was examined through chemical and mechanical stimulation, and open field test measured the anxiety levels. Restingstate fMRI is a method of functional magnetic resonance imaging (rs-fMRI) was performed for anatomical involvement of the brain regions. The amplitude of low-frequency fluctuation (ALFF) determined brain activity. Immunofluorescence testing and Quantitative real-time polymerase chain reaction were also performed to further validate the findings. Compared with the Sham group, ALFF signals in the supplemental somatosensory area, secondary auditory cortex, agranular insular cortex, temporal association areas, and ectorhinal cortex brain areas were increased in the dry eye group. This change of ALFF in the insular cortex was linked with the increment in corneal hypersensitivity (p < 0.01), c-Fos (p < 0.001), brain-derived neurotrophic factor (p < 0.01), TNF-α, IL-6, and IL-1ß (p < 0.05). In contrast, IL-10 levels (p < 0.05) decreased in the dry eye group. DED-induced corneal hypersensitivity and upregulation of inflammatory cytokines could be blocked by insular cortex injection of Tyrosine Kinase receptor B agonist cyclotraxin-B (p < 0.01) without affecting anxiety levels. Our study reveals that the functional activity of the brain associated with corneal neuropathic pain and neuroinflammation in the insular cortex might contribute to dry eye-related corneal neuropathic pain.

Representation, mining and analysis of unsafe behaviour based on pan-scene data.

To describe the safety rules of various industrial process data and explore the characteristics of unsafe behaviour, the association rules of unsafe behaviour based on pan-scene were proposed in this study. First, based on the scene data theory, unsafe behaviour was described by eight dimensions (time, location, behavioural individual, unsafe action, behavioural attribute, behavioural trace, professional category and risk level) to achieve scene data description and structural transformation. Second, the Apriori algorithm was used to explore the distribution rules of unsafe behaviour dimensions and the interaction between different dimensions from two perspectives: single-dimensional statistical analysis and multidimensional association rule mining. Finally, through SPSS Modeler software, an empirical analysis of pan-scene data for subway construction was conducted, and the association rules between type of work, construction stage, working time and unsafe action were identified. Some strong association rules were produced by the association analysis. For example, during the 13:00-17:00 of the excavation floor stage, the most frequent unsafe action of machine operators is the irregular binding of lifting objects. This result could explain why some unsafe actions are prone to occur in different construction stages and working times for workers of different types, which can be controlled and managed in a targeted manner, thus reducing the possibility of accidents.

Selective adsorption of various phosphorus species coexistence in water-soluble ammonium polyphosphate on goethite: Experimental investigation and molecular dynamics simulation.

The geochemical processes of polyphosphates (poly-Ps) are important for phosphorus (P) management and environmental protection. Water-soluble ammonium polyphosphate (APP) containing various P species has been increasingly used as an alternative P-fertilizer. The various P species coexistence and the chelation of poly-Ps with mental would trigger the P's competitive adsorption and affect the APP's adsorption intensity on goethite, compared to single orthophosphate (P1). P adsorption behaviors of APP1 with two P species and APP2 with seven P species on goethite were investigated via batch experiments in comparison to the traditional P-fertilizer of mono-ammonium phosphate (MAP). Coadsorption of P1 and pyrophosphate (P2) on goethite was investigated by molecular dynamics (MD) simulation. The more Fe3+ dissolved from goethite as a bridge due to the chelation of poly-Ps in APP and contributed to the stronger APP adsorption on goethite compared with MAP. Ion chromatography and spectral analysis showed P1 and P2 in APP were mainly adsorbed by goethite via mainly forming bidentate complexes. The goethite preferentially adsorbed P1 at lower APP concentration but increased the poly-Ps' adsorption at higher APP concentration. MD simulation showed that electrostatic interaction and hydrogen bonds played a key role in water-phosphates-goethite systems. The P1 pre-adsorbed on goethite could be replaced by P2 at high P2 concentration. The results develop new insights regarding the selective adsorption of various P species coexistence in goethite-rich environments.

Spatio-temporal variation of net anthropogenic nitrogen inputs (NANI) from 1991 to 2019 and its impacts analysis from parameters in Northwest China.

At present, excessive nutrient inputs caused by human activities have resulted in environmental problems such as agricultural non-point source pollution and water eutrophication. The Net Anthropogenic Nitrogen Inputs (NANI) model can be used to estimate the nitrogen (N) inputs to a region that are related to human activities. To explore the net nitrogen input of human activities in the main grain-producing areas of Northwestern China, the county-level statistical data for the Ningxia province and NANI model parameters were collected, the spatio-temporal distribution characteristics of NANI were analyzed and the uncertainty and sensitivity of the parameters for each component of NANI were quantitatively studied. The results showed that: (1) The average value of NANI in Ningxia from 1991 to 2019 was 7752 kg N km-2 yr-1. Over the study period, the inputs first showed an overall increase, followed by a decrease, and then tended to stabilize. Fertilizer N application was the main contributing factor, accounting for 55.6%. The high value of NANI in Ningxia was mainly concentrated in the Yellow River Diversion Irrigation Area. (2) The 95% confidence interval of NANI obtained by the Monte Carlo approach was compared with the results from common parameters in existing literature. The simulation results varied from -6.4% to 27.4% under the influence of the changing parameters. Net food and animal feed imports were the most uncertain input components affected by parameters, the variation range was -20.7%-77%. (3) The parameters of inputs that accounted for higher proportions of the NANI were more sensitive than the inputs with lower contributions. The sensitivity indexes of the parameters contained in the fertilizer N applications were higher than those of net food and animal feed imports and agricultural N-fixation. This study quantified the uncertainty and sensitivity of parameters in the process of NANI simulation and provides a reference for global peers in the application and selection of parameters to obtain more accurate simulation results.

Enhanced phosphorus mobility in a calcareous soil with organic amendments additions: Insights from a long term study with equal phosphorus input.

The agricultural practice of replacing chemical fertilizers with organic amendments (manure and/or straw) may have consequences for phosphorus (P) loss to the environment. Such a knowledge gap was examined using a ten-year field trial in calcareous soil containing four treatments with the equal annual P input but varied organic amendment combinations as follows: mineral fertilizer only as control (MF), mineral fertilizer coupled with manure (MM), mineral fertilizer coupled with manure and straw (MMS) and mineral fertilizer coupled with straw (MS). The soil P distribution, P fractions and speciation, Fe(III) reduction and P sorption kinetics were investigated using the chemical extraction, K edge X-ray absorption near-edge structure and Langmuir equations. The electronic shuttle capacity of soils and speciation of soil dissolved organic matter (DOM) were also evaluated using electrochemical methods, three-dimensional excitation-emission matrix fluorescence spectroscopy and Fourier transform infrared spectra methods. Results showed that soil Olsen-P and total P increased at depths of 20-40 cm in MM, MMS and MS treatments, suggesting that manure and/or straw addition significantly mobilized P in the soil profile. Manure and/or straw addition also decreased soil maximum P sorption capacity (Smax) and increased the desorption rate at depths of 0-20 cm in soil across treatments. At a depth of 0-20 cm in soil of the MS treatment, the enhanced Fe(Ⅲ) reduction coupled with a decrease of Fe-bound P supports that Fe reduction dominates the mobilization of P. The transformation of Ca bound-P to Al/Fe bound-P in a depth of 0-20 cm in soil of the MM treatment may be due to the high proportion of humic-like substances in the DOM at a depth of 0-20 cm in soil of the MM treatment, which may have caused a slight/microsite acidification. These results can help to develop optimized fertilization practices to effectively mitigate P loss from calcareous soils with manure and/or straw addition.

Investigation of differential levels of phosphorus fixation in dolomite and calcium carbonate amended red soil.

BACKGROUND: The pH adjustment of acidic red soils with lime materials is beneficial for the reduction of phosphorus (P) fixation. However, the reasons for varying levels of P activation after adding different lime materials have not been fully investigated. Therefore, this study examined changes in soil labile P and P forms after phosphate application to calcium carbonate (CaCO3 ) and dolomite amended red soil during a 120-day incubation period. Also change of P sorption properties in the amended soil samples from day 120 were examined through a sorption-desorption experiment. RESULTS: The increase of soil H2 O-P and NaHCO3 -P in the CaCO3 and dolomite amended soil treatments was mainly ascribed to the decline of the NaOH-P. However, when compared with the control treatment after 120 days, soil Olsen-P significantly increased by 34% and 66% in the CaCO3 and dolomite treatments. The Hedley P fractionation results demonstrated that the CaCO3 application caused a notable increase of HCl-P (stable Ca-P), which was 88.4% higher than that in the dolomite treatment. However, the formation of stable P was strongly suppressed in the dolomite treatment due to the presence of magnesium (Mg), which was identified by the negative relationship between M3-Mg and HCl-P. In line with these findings, P sorption-desorption work showed weaker P binding energy in the dolomite treatment relative to the CaCO3 treatment. CONCLUSION: In terms of increasing P availability in red soil, this study suggests that dolomite should be used to substitute CaCO3 in order to reduce the soil P fixation. © 2021 Society of Chemical Industry.

Leached phosphorus apportionment and future management strategies across the main soil areas and cropping system types in northern China.

Excess phosphorus (P) leached from high fertiliser input cropping systems in northern China is having detrimental effects on water quality. Before improved management can be directed at specific soils and cropping system types estimates of P leached loss apportionment and mitigation potentials across the main soil (fluvo-aquic soil, FAS; cinnamon soil, CS; black soil, BS) areas and cropping systems (protected vegetable fields, PVFs; open vegetable fields, OVFs; cereal fields, CFs) are needed. The present study designed and implemented conventional fertilisation and low input system trials at 75 sites inclusive of these main soils and cropping system types in northern China. At all sites, a uniform lysimeter design (to 0.9 m depth) enabled the collection and analysis of leachate samples from 7578 individual events between 2008 and 2018. In addition, site-specific static and dynamic activity data were recorded. Results showed that annual total phosphorus (TP) leached losses across the main soil areas and cropping systems were 4.99 × 106 kg in northern China. A major finding was PVFs contributed to 48.5% of the TP leached losses but only accounted for 5.7% of the total cropping areas. The CFs and OVFs accounted for 40.3% and 11.2% of the TP leached losses, respectively. Across northern China, the TP leached losses in PVFs and OVFs were greatest in FAS areas followed by CS and BS areas. The higher TP leached losses in FAS areas were closely correlated with greater P fertiliser inputs and irrigation practices. From a management perspective in PVFs and OVFs systems, a decrease of P inputs by 10-30% would not negatively affect yields while protecting water quality. The present study highlights the importance of decreasing P inputs in PVFs and OVFs and supporting soil P nutrient advocacy for farmers in China.

Elucidation and Regulation of Polyphenols in the Smoking Process of Shanxi Aged Vinegar.

Polyphenols (PPs) are the main contributors to the health functions of Shanxi aged vinegar (SAV) and are mainly produced during the smoking process. This study aimed to explore the feasibility of regulating the accumulation of total water-soluble PPs (TWSP) by changing environmental factors based on the distribution of PPs. A total of eleven PPs, such as vanillin, vanillic acid, and (e)-ferulic acid, were detected during the smoking process. During the smoking process, the content of TWSP gradually increased and was accompanied by changes in environmental factors. Spearman correlation analysis and verification experiments showed that temperature, amino acids, and reducing sugars, as the main influencing factors, promoted the accumulation of TWSP. The in situ regulation strategy of changing environmental factors significantly increased the accumulation of TWSP by 12.24%.

Behaviors Related to Psychiatric Disorders and Pain Perception in C57BL/6J Mice During Different Phases of Estrous Cycle.

Robust sex difference among humans regarding psychiatry- and pain-related behaviors is being researched; however, the use of female mice in preclinical research is relatively rare due to an unchecked potential behavioral variation over the estrous cycle. In the present study, a battery of psychiatry- and pain-related behaviors are examined under physiological condition in female C57BL/6J mice over different estrous cycle phases: proestrus, estrous, metestrous, diestrous. Our behavioral results reveal that there is no significant difference over different phases of the estrous cycle in social interaction test, sucrose preference test, tail suspension test, open field test, marble burying test, novelty-suppressed feeding test, Hargreaves thermal pain test, and Von Frey mechanical pain test. These findings implicate those psychiatry- and pain-related behaviors in normal female C57BL/6J mice appear to be relatively consistent throughout the estrous cycle; the estrous cycle might not be a main contributor to female C57BL/6J mice's variability of behaviors.

Activation of anterior thalamic reticular nucleus GABAergic neurons promotes arousal from propofol anesthesia in mice.

Propofol is widely used for the induction and maintenance of anesthesia, which causes a rapid loss of consciousness. However, the mechanisms underlying the hypnosis effect of propofol are still not fully understood. The thalamic reticular nucleus (TRN) is crucial for regulating wakefulness, sleep rhythm generation, and sleep stability, while the role of TRN in the process of propofol-induced anesthesia is still unknown. Here, we investigated the function of the anterior TRN in propofol general anesthesia. Our results demonstrated that the neural activity of anterior TRN is suppressed during propofol anesthesia, whereas it is robustly activated from anesthesia by recording the calcium signals using fiber photometry technology. The results showed that the activation of anterior TRN neurons by chemogenetic and optogenetic methods shortens the emergency time without changing the induction time. Conversely, chemogenetic or optogenetic inhibition of the TRN neurons leads to a delay in the recovery time. Our study showed that anterior TRN is crucial for behavioral arousal without affecting the induction time of propofol anesthesia.

Identification of an Intermediate Form of Ferredoxin That Binds Only Iron Suggests That Conversion to Holo-Ferredoxin Is Independent of the ISC System in Escherichia coli.

Escherichia coli [2Fe-2S]-ferredoxin and other ISC proteins encoded by the iscRSUA-hscBA-fdx-iscX (isc) operon are responsible for the assembly of iron-sulfur clusters. It is proposed that ferredoxin (Fdx) donates electrons from its reduced [2Fe-2S] center to iron-sulfur cluster biogenesis reactions. However, the underlying mechanisms of the [2Fe-2S] cluster assembly in Fdx remain elusive. Here, we report that Fdx preferentially binds iron, but not the [2Fe-2S] cluster, under cold stress conditions (≤16°C). The iron binding in Fdx is characterized by a unique absorption peak at 320 nm based on UV-visible spectroscopy. In addition, the iron-binding form of Fdx could be converted to the [2Fe-2S] cluster-bound form after transferring cold-stressed cells to normal cultivation temperatures above 25°C. In vitro experiments also revealed that Fdx could utilize bound iron to assemble the [2Fe-2S] cluster by itself. Furthermore, inactivation of the genes encoding IscS, IscU, and IscA did not limit [2Fe-2S] cluster assembly in Fdx, which was also observed by inactivating the isc or suf operon, indicating that iron-sulfur cluster biogenesis in Fdx arose from a unique pathway in E. coli Our results suggest that the intracellular assembly of [2Fe-2S] clusters in Fdx is susceptible to environmental temperatures. The iron binding form of Fdx (Fe-Fdx) is a precursor during its maturation to a cluster binding form ([2Fe-2S]-Fdx), and reassembly of the [2Fe-2S] clusters during temperature increases is not strictly reliant on other specific iron donors and scaffold proteins within the Isc or Suf system.IMPORTANCE Fdx is an electron carrier that is required for the maturation of many other iron-sulfur proteins. Its function strictly depends on its [2Fe-2S] center that bonds with the cysteinyl S atoms of four cysteine residues within Fdx. However, the assembly mechanism of the [2Fe-2S] clusters in Fdx remains controversial. This study reports that Fdx fails to form its [2Fe-2S] cluster under cold stress conditions but instead binds a single Fe atom at the cluster binding site. Moreover, when temperatures increase, Fdx can assemble clusters by itself from its iron-only binding form in E. coli cells. The possibility remains that Fdx can effectively accept clusters from multiple sources. Nevertheless, our results suggest that Fdx has a strong iron binding activity that contributes to the assembly of its own [2Fe-2S] cluster and that Fdx acts as a temperature sensor to regulate Isc system-mediated iron-sulfur cluster biogenesis.

Alum split applications strengthened phosphorus fixation and phosphate sorption in high legacy phosphorus calcareous soil.

High phosphorus (P) saturation arising from historic P inputs to protected vegetable fields (PVFs) drives high P mobilisation to waterbodies. Amendment of soils with alum has shown potential in terms of fixing labile P and protecting water quality. The present 15 month pot experiment investigated P stabilisation across single alum application (Alum-1 treatment, 20 g alum/kg soil incorporated into soil before the maize was sown), alum split applications (Alum-4 treatment, 5 g alum/kg soil incorporated into soil before each crop was sown i.e. 4 × 5 g/kg) and soil only treatment (Control). Results showed that the Alum-1 treatment caused the strongest stabilisation of soil labile P after maize plant removal, whereas the P stabilisation effect was gradually weakened due to the transformation of soil non-labile P to labile P and the reduced active Al3+ in soil solution. For the Alum-4 treatment, soil labile P decreased gradually with each crop planting and was lower than the Alum-1 treatment at the end of the final crop removal, without any impairment on plant growth. The better P stabilisation at the end of Alum-4 treatment was closely correlated with a progressive supply of Al3+ and a gradual decrease of pH, which resulted in higher contents of poorly-crystalline Al, Fe and exchangeable Ca. These aspects were conducive to increasing the soil P stabilisation and phosphate sorption. In terms of management, growers in continuous cropping systems could utilise split alum applications as a strategy to alleviate P losses in high-P enriched calcareous soil.

Removal of Chromium (VI) by Escherichia coli Cells Expressing Cytoplasmic or Surface-Displayed ChrB: a Comparative Study.

Various genetically engineered microorganisms have been developed for the removal of heavy metal contaminants. Metal biosorption by whole-cell biosorbents can be enhanced by overproduction of metal-binding proteins/peptides in the cytoplasm or on the cell surface. However, few studies have compared the biosorption capacity of whole cells expressing intracellular or surface-displayed metal-adsorbing proteins. In this study, several constructs were prepared for expressing intracellular and surface-displayed Ochrobactrum tritici 5bvl1 ChrB in Escherichia coli BL21(DE3) cells. E. coli cells expressing surface-displayed ChrB removed more Cr(VI) from aqueous solutions than cells with cytoplasmic ChrB under the same conditions. However, intracellular ChrB was less susceptible to variation in extracellular conditions (pH and ionic strength), and more effectively removed Cr(VI) from industrial wastewater than the surface-displayed ChrB at low pH (<3). An adsorptiondesorption experiment demonstrated that compared with intracellular accumulation, cell-surface adsorption is reversible, which allows easy desorption of the adsorbed metal ions and regeneration of the bioadsorbent. In addition, an intrinsic ChrB protein fluorescence assay suggested that pH and salinity may influence the Cr(VI) adsorption capacity of ChrB-expressing E. coli cells by modulating the ChrB protein conformation. Although the characteristics of ChrB may not be universal for all metal-binding proteins, our study provides new insights into different engineering strategies for whole-cell biosorbents for removing heavy metals from industrial effluents.

Understanding phosphate sorption characteristics of mineral amendments in relation to stabilising high legacy P calcareous soil.

In China, excessive phosphorus (P) application in protected vegetable fields has led to high legacy P stores. Soil amendment with alum or dolomite is one of many best management practices (BMPs) used to reduce P losses in calcareous soils. However, both the kinetics and mechanisms of P sorption and soil available P in amended soils are understudied. Herein, both aspects were looked at under controlled conditions. Firstly, a sorption study which coupled P concentrations with poorly-crystalline Al hydroxides and dolomite was conducted. Results from this batch experiment showed that P sorption on poorly-crystalline Al hydroxides was homogenous and occurred mainly via displacement of inner-sphere hydroxyl (Al-OH) instead of the formation of AlPO4. However, the amount of sorbed P reached maximum sorption of 73.1 mg g-1 and did not change with further increase in P concentration. It was observed that P adsorbed onto the dolomite surface at low P concentrations, whereas hydroxyl replacement and uneven cluster precipitation of Ca3(PO4)2 occurred at high P concentrations. A second 90 day incubation experiment investigated changes to soil available P and sorption-desorption across variable rates of amendments (0-50 g kg-1). Results showed that alum amendment at a rate of 50 g kg-1 decreased soil CaCl2-P and Olsen-P concentrations by 91.9% and 57.8%, respectively. However, Olsen-P increased when the dolomite rates were <20 g kg-1. Phosphorus sorption-desorption of the amended soil showed alum had higher P sorption efficiency than dolomite at low addition rates (<10 g kg-1). However, soil amended with high dolomite rates (>10 g kg-1) could sorb more P in comparison with alum when P concentrations were increased. The P status of the amended soil was closely connected to the P sorption mechanisms on mineral amendments, soil P concentrations and soil properties.