Total and denitrifying bacterial communities associated with the interception of nitrate leaching by carbon amendment in the subsoil.

Nitrate leaching is severe in greenhouse where excessive nitrogen is often applied to maintain high crop productivities. In this study, we investigated the effects of carbon amendment in the subsoil on nitrate leaching and the emission of greenhouse gases (CH4 and N2O) using a soil column experiment. Carbon amendment resulted in over 39% reduction in nitrate leaching and 25.3% to 60.6% increase of total N content in the subsoil zone as compared to non-amended control. Strikingly, the abundance of nirS, nosZ, and 16S rRNA were higher in the treatment than the corresponding controls while no significant effect was detected for nirK. Carbon amendment explained 14%, 10%, and 4% of the variation in the community of nosZ, nirS, and nirK, respectively. It also considerably (more than 7 times) enriched genera such as Anaerovorax, Pseudobacteroides, Magnetospirillum, Prolixibacter, Sporobacter, Ignavibacterium, Syntrophobacter, Oxobacter, Hydrogenispora, Desulfosporomusa, Mangrovibacterium, and Sporomusa, as revealed by the analysis of 16S rRNA amplicon. Network analysis further uncovered that carbon amendment enriched three microbial hubs which mainly consists of positively correlated nirS, nosZ, and anaerobic bacterial populations. In summary, carbon amendment in the subsoil mitigated nitrate leaching and increased the nitrogen pool by possible activation of denitrifying and anaerobic bacterial populations. KEY POINTS: • Carbon amendment in subsoil reduced NO3- leaching by over 39% under high N input. • Carbon amendment increased the total N in subsoil from 25.3% to 60.6%. • Carbon amendment enriched nirS- and nosZ-type denitrifying bacteria in subsoil.

Different functional susceptibilities of mouse retinal ganglion cell subtypes to optic nerve crush injury.

In optic neuropathies, the progressive deterioration of retinal ganglion cell (RGC) function leads to irreversible vision loss. Increasing experimental evidence suggests differing susceptibility for RGC functional subtypes. Here with multi-electrode array recordings, RGC functional loss was characterized at multiple time points in a mouse model of optic nerve crush. Firing rate, latency of response and receptive field size were analyzed for ON, OFF and ON-OFF RGCs separately. It was observed that responses and receptive fields of OFF cells were impaired earlier than ON cells after the injury. For the ON-OFF cells, the OFF component of response was also more susceptible to optic nerve injury than the ON component. Moreover, more ON transient cells survived than ON sustained cells post the crush, implying a diversified vulnerability for ON cells. Together, these data support the contention that RGCs' functional degeneration in optic nerve injury is subtype dependent, a fact that needs to be considered when developing treatments of glaucomatous retinal ganglion cell degeneration and other optic neuropathies.

[Information coding in retinal ganglion cells].

In vertebrate visual system, retina is the first stage for visual information processing. Retinal ganglion cells are the only output neurons of the retina, and their firing activities are dependent on visual stimuli. Retinal ganglion cells can effectively encode visual information via various manners, such as firing rate, temporal structure of spike trains, and concerted activity, etc. Adaptation is one of the basic characteristics of the nervous system, which enables retinal neurons to encode stimuli under a wide variety of natural conditions with limited range in their output. This article reviews the recent studies focused on the coding properties and adaptation of retinal ganglion cells. Relevant issues about dynamical adjustment of coding strategies of retinal ganglion cells in response to different visual stimulation, as well as physiological property and function of adaptation are discussed.

Gap junction permeability modulated by dopamine exerts effects on spatial and temporal correlation of retinal ganglion cells' firing activities.

Synchronized activities among retinal ganglion cells (RGCs) via gap junctions can be increased by exogenous dopamine (DA). During DA application, single neurons' firing activities become more synchronized with its adjacent neighbors. One intriguing question is how the enhanced spatial synchronization alters the temporal firing structure of single neurons. In the present study, firing activities of bullfrog's dimming detectors in response to binary pseudo-random checker-board flickering were recorded via a multi-channel recording system. DA was applied in the retina to modulate synchronized activities between RGCs, and the effect of DA on firing activities of single neurons was examined. It was found that, during application of DA, synchronized activities between single neuron and its neighboring neurons was enhanced. At the meantime, the temporal structures of single neuron spike train changed significantly, and the temporal correlation in single neuron's response decreased. The pharmacological study results indicated that the activation of D1 receptor might have effects on gap junction permeability between RGCs. Our results suggested that the dopaminergic pathway participated in the modulation of spatial and temporal correlation of RGCs' firing activities, and may exert critical effects on visual information processing in the retina.

Effect of the entorhinal cortex on ictal discharges in low-Mg²âº-induced epileptic hippocampal slice models.

The hippocampus plays an important role in the genesis of mesial temporal lobe epilepsy, and the entorhinal cortex (EC) may affect the hippocampal network activity because of the heavy interconnection between them. However, the mechanism by which the EC affects the discharge patterns and the transmission mode of epileptiform discharges within the hippocampus needs further study. Here, multielectrode recording techniques were used to study the spatiotemporal characteristics of epileptiform discharges in adult mouse hippocampal slices and combined EC-hippocampal slices and determine whether and how the EC affects the hippocampal neuron discharge patterns. The results showed that low-Mg²âº artificial cerebrospinal fluid induced interictal discharges in hippocampal slices, whereas, in combined EC-hippocampal slices the discharge pattern was alternated between interictal and ictal discharges, and ictal discharges initiated in the EC and propagated to the hippocampus. The pharmacological effect of the antiepileptic drug valproate (VPA) was tested. VPA reversibly suppressed the frequency of interictal discharges but did not change the initiation site and propagation speed, and it completely blocked ictal discharges. Our results suggested that EC was necessary for the hippocampal ictal discharges, and ictal discharges were more sensitive than interictal discharges in response to VPA.

Involvement of thalamus in initiation of epileptic seizures induced by pilocarpine in mice.

Studies have suggested that thalamus is involved in temporal lobe epilepsy, but the role of thalamus is still unclear. We obtained local filed potentials (LFPs) and single-unit activities from CA1 of hippocampus and parafascicular nucleus of thalamus during the development of epileptic seizures induced by pilocarpine in mice. Two measures, redundancy and directionality index, were used to analyze the electrophysiological characters of neuronal activities and the information flow between thalamus and hippocampus. We found that LFPs became more regular during the seizure in both hippocampus and thalamus, and in some cases LFPs showed a transient disorder at seizure onset. The variation tendency of the peak values of cross-correlation function between neurons matched the variation tendency of the redundancy of LFPs. The information tended to flow from thalamus to hippocampus during seizure initiation period no matter what the information flow direction was before the seizure. In some cases the information flow was symmetrically bidirectional, but none was found in which the information flowed from hippocampus to thalamus during the seizure initiation period. In addition, inactivation of thalamus by tetrodotoxin (TTX) resulted in a suppression of seizures. These results suggest that thalamus may play an important role in the initiation of epileptic seizures.

Promoting agricultural waste-driven denitrification and nitrogen sequestration with nano-enabled strategy.

Nanotechnology and biotechnology offer promising avenues for bolstering food security through the facilitation of soil nitrogen (N) sequestration and the reduction of nitrate leaching. Nonetheless, a comprehensive and mechanistic evaluation of their effectiveness and safety remains unclear. In this study, a soil remediation strategy employing nano-Fe3O4 and straw in N-contaminated soil was developed to elucidate N retention mechanisms via diverse metagenomics techniques. The findings revealed that subsoil amended with straw, particularly in conjunction with nano-Fe3O4, significantly increased subsoil N content (53.2%) and decreased nitrate concentration (74.6%) in leachate. Furthermore, the enrichment of functional genes associated with N-cycling, sulfate, nitrate, and iron uptake, along with chemotaxis, and responses to environmental stimuli or microbial collaboration, effectively mitigates nitrate leaching while enhancing soil N sequestration. This study introduces a pioneering approach utilizing nanomaterials in soil remediation, thereby offering the potential for the cultivation of safe vegetables in high N input greenhouse agriculture.

A microfluidic liquid phase nucleic acid purification chip to selectively isolate DNA or RNA from low copy/single bacterial cells in minute sample volume followed by direct on-chip quantitative PCR assay.

Purification of nucleic acids from a low quantity of bacterial cells in minute volume is important in many clinical and biological applications. We developed a novel microfluidic liquid phase nucleic acid purification chip to selectively isolate DNA or RNA from bacterial cells in the range of 5000 down to a single cell in the sample volume of 1 µl or 125 nl, which can be directly put through on-chip quantitative PCR assay. The aqueous phase bacterial lysate was isolated in an array of microwells, after which an immiscible organic (phenol-chloroform) phase was introduced in a headspace channel connecting the microwell array. Continuous flow of the organic phase increases the interfacial contact with the aqueous phase to achieve purification of target nucleic acid through phase partitioning. Significantly enhanced nucleic acid recovery yield, up to 10 fold higher, was achieved using the chip-based liquid phase nucleic acid purification technique compared to that obtained by the conventional column-based solid phase nucleic acid extraction method. One step vacuum-driven microfluidics allowed an on-chip quantitative PCR assay to be carried out in the same microwells within which bacterial nucleic acids were isolated, avoiding sample loss during liquid transfer. Using this nucleic acid purification device set in a two-dimensional (2D) array format of 900 microwells, it was demonstrated for the first time that high-throughput extraction of RNA couple with direct on-chip PCR analysis from single bacterial cells could be achieved. Our microfluidic platform offered a simple and effective solution for nucleic acid preparation, which can be integrated for automated bacterial pathogen detection and high throughput transcriptional profiling.

A high-throughput microfluidic biochip to quantify bacterial adhesion to single host cells by real-time PCR assay.

A high-throughput microfluidic poly-(dimethylsiloxane) biochip was developed to quantify bacterial adhesion to single host cells by real-time PCR assay. The biochip is simply structured with a two-dimensional array of 900 micro-wells, one inlet, and one outlet micro-channels. Isolation of single infected host cells into the individual micro-wells of the biochip was achieved by one-step vacuum-driven microfluidics. The adhered bacterial cells were then quantified by direct on-chip real-time PCR assay with single-bacterium-detection sensitivity. The performance of this microfluidic platform was demonstrated through profiling of the association of a common bacterial pathogen, Pseudomonas aeruginosa, to single host human lung epithelial A549 cells, revealing an adherence distribution that has not been previously reported. This microfluidic platform offers a simple and effective tool for biologists to analyze pathogen-host interaction at the single-cell level without the necessities of fluorescence labeling. The chip can similarly be used for other PCR-based applications requiring single-cell analysis.

Predefinable colorimetric quantum-dot barcodes with simple and express identification algorithm.

Specific fluorescent profiles were created by loading quantum-dot (Qdot) mixtures in liquid cores of monodispersed polymer microcapsules, which were used as colorimetric barcodes for small object identification. Since the emission intensities of Qdot-loaded liquid cores maintain a linear relation to the Qdot concentrations and the Qdots in the liquid cores with different emission peaks have no obvious interference, the colorimetric barcodes can be predefined by the compositions of Qdot mixtures. The colorimetric barcodes can be identified easily by recording the emission intensities of the encoded microcapsules at respective Qdot emission peaks with a simple and express algorithm, which are suitable to conduct high-throughput multiplexed assays by flow cytometer for biological screening applications.

Mechanism of low-frequency, low-intensity ultrasound modulation of the mouse retina.

Objective.Ultrasound has been shown to modulate the activity of retinal ganglion cells (RGCs) in mice, but the mechanism remains poorly understood. This study aims to address this question.Approach.Multi-electrode recordings together with pharmacological methods were used to investigate the possible cellular/circuitry mechanism(s) underlying the neuronal modulation induced by low-frequency (1 MHz), low-intensity (ISPTA0.5 W cm-2) ultrasound stimulation.Main results.We found that ultrasound activated mechanosensitive channels (transient receptor potential vanilloid 4 (TRPV4) channels are involved) in Müller cells, causing the release of glutamate, which acts on the extrasynapticN-methyl-D-aspartate receptors of RGCs, thus leading to the modulation of neuronal activity.Significance.Our results reveal a novel mechanism of low-frequency, low-intensity ultrasound modulation, involving TRPV4 as a mechanosensitive target for ultrasound and glutamate as an essential mediator of neuron-glia communication. These findings also demonstrate that the mechanical-force-mediated pathway is important for retinal signal modulation during visual processes, such as visual accommodation.

Getting a Foothold on Diabetic Foot Disease-Outcomes of a Multidisciplinary Clinical Pathway for Inpatient Diabetic Foot Care: A 17-Year Institutional Review.

INTRODUCTION: Diabetes foot disease (DFD) contributes to poor quality of life, clinical and economic burden. Multidisciplinary diabetes foot teams provide prompt access to specialist teams thereby improving limb salvage. We present a 17-year review of an inpatient multidisciplinary clinical care path (MCCP) for DFD in Singapore. METHODS: This was a retrospective cohort study of patients admitted for DFD and enrolled in our MCCP to a 1700-bed university hospital from 2005 to 2021. RESULTS: There were 9279 patients admitted with DFD with a mean of 545 (±119) admissions per year. The mean age was 64 (±13.3) years, 61% were Chinese, 18% Malay and 17% Indian. There was a higher proportion of Malay (18%) and Indian (17%) patients compared to the country's ethnic composition. A third of the patients had end stage renal disease and prior contralateral minor amputation. There was a reduction in inpatient major lower extremity amputation (LEA) from 18.2% in 2005 to 5.4% in 2021 (odds ratio 0.26, 95% confidence interval 0.16-0.40, P < .001) which was the lowest since pathway inception. Mean time from admission to first surgical intervention was 2.8 days and mean time from decision for revascularization to procedure was 4.8 days. The major-to-minor amputation rate reduced from 1.09 in 2005 to 0.18 in 2021, reflecting diabetic limb salvage efforts. Mean and median length of stay (LOS) for patients in the pathway was 8.2 (±14.9) and 5 (IQR = 3) days, respectively. There was a gradual trend of increase in the mean LOS from 2005 to 2021. Inpatient mortality and readmission rate was stable at 1% and 11%. CONCLUSION: Since the institution of a MCCP, there was a significant improvement in major LEA rate. An inpatient multidisciplinary diabetic foot care path helped to improve care for patients with DFD.

A high-flux isopore micro-fabricated membrane for effective concentration and recovering of waterborne pathogens.

A high-flux metallic micro/nano-filtration membrane has been fabricated and validated for isolation of waterborne pathogens from drinking water. Obtained membrane with smooth surface and perfectly ordered pores was achieved by a high yield and cost effective multilevel lithography and electroplating technique. The micro-fabricated membrane was also strengthened with an integrated back-support, which can withstand a high pressure during filtration. The results of microfiltration tests with model particles revealed the superior performance of the micro-fabricated filter than current commercial filters in sample throughput, recovery ratio, and reusability. This study highlighted the potential application of micro-fabricated filer in rapid filtration and recovery of C. parvum oocysts for downstream analysis.

Complementary effects of phosphorus supply and planting density on maize growth and phosphorus use efficiency.

Phosphorus (P) supply and planting density regulate plant growth by altering root morphological traits and soil P dynamics. However, the compensatory effects of P supply and planting density on maize (Zea mays L.) growth and P use efficiency remain unknown. In this study, we conducted pot experiments of approximately 60 days to determine the effect of P supply, i.e., no P (CK), single superphosphate (SSP), and monoammonium phosphate (MAP), and different planting densities (low: two plants per pot; and high: four plants per pot) on maize growth. A similar shoot biomass accumulation was observed at high planting density under CK treatment (91.5 g plot-1) and low planting density under SSP treatment (94.3 g plot-1), with similar trends in P uptake, root morphological traits, and arbuscular mycorrhizal colonization. There was no significant difference in shoot biomass between high planting density under SSP (107.3 g plot-1) and low planting density under MAP (105.2 g plot-1); the corresponding P uptake, root growth, and P fraction in the soil showed the same trend. These results suggest that improved P supply could compensate for the limitations of low planting density by regulating the interaction between root morphological traits and soil P dynamics. Furthermore, under the same P supply, the limitations of low planting density could be compensated for by substituting MAP for SSP. Our results indicate that maize growth and P use efficiency could be improved by harnessing the compensatory effects of P supply and planting density to alter root plasticity and soil P dynamics.

Using knowledge-based management for sustainable phosphorus use in China.

Sustainable phosphorus (P) management presents challenges in crop production and environmental protection; the current understanding of chemical P-fertilizer manufacturing, rock phosphate (RP) mining, P loss within supply chains, and strategies to mitigate loss is incomplete because of a fragmented understanding of P in the crop production supply chain. Therefore, we develop a knowledge-based management theoretical framework to analyze P supply chains to explore ways to mitigate China's P crisis. This framework connects upstream P industries and crop production, addressing knowledge gaps and stakeholder involvement. We demonstrate the potential to improve P use efficiency in the supply chain, thereby mitigating the P crisis using optimized P management. Our results showed that P footprint and grain production demand for RP can be reduced without yield penalty using a crop-demand-oriented P supply chain management that integrates P use in crop production, P-fertilizer manufacturing, and RP mining. Food security and P-related environment sustainability can be achieved by sharing responsibility and knowledge among stakeholders.

Synergies in sustainable phosphorus use and greenhouse gas emissions mitigation in China: Perspectives from the entire supply chain from fertilizer production to agricultural use.

Synergies to achieve high phosphorus (P) use efficiency (PUE) and mitigate greenhouse gas (GHG) emissions are critical for developing strategies aimed toward agricultural green development. However, the potential effects of such synergies in the entire P supply chain through optimizing P management in crop production are poorly understood. In this study, a partial life cycle of a GHG emissions model was developed to quantify the P-related GHG emissions in the entire P supply chain in China. Our results showed that 16.3 kg CO2-equivalent (CO2-eq) was produced from the entire P supply chain per unit of P used for grain agriculture (maize, rice, and wheat). P-related GHG emissions in China increased more than five-fold from 1980 (7.2 Tg CO2-eq) to 2018 (44.9 Tg CO2-eq). GHG emissions were found to be strongly associated with the intensity of grain production in China, and they varied considerably across production regions owing to the differences in the P fertilizer production efficiency. Mineral P fertilizer use in crop production was the primary source of P-related GHG emissions. The results suggest that sustainable P management by matching mineral P fertilizer rates and fertilizer types with crop needs can mitigate GHG emissions by 10.8-27.7 Tg (24.0-65.1%). Moreover, this can improve PUE and reduce mineral P input by 0.7-1.4 Tg (24.0-46.0%). These findings highlight that potential synergies between high PUE and low P-related GHG emissions can be achieved via sustainable P management, thereby enhancing green agricultural development in China and other regions worldwide.

Low-frequency, low-intensity ultrasound modulates light responsiveness of mouse retinal ganglion cells.

Objective. Ultrasound modulates the firing activity of retinal ganglion cells (RGCs), but the effects of lower-frequency, lower-intensity ultrasound on RGCs and underlying mechanism(s) remain poorly understood. This study aims to address these questions.Approach. Multi-electrode recordings were used in this study to record the firing sequences of RGCs in isolated mouse retinas. RGCs' background firing activities as well as their light responses were recorded with or without ultrasound stimulation. Cross-correlation analyses were performed to investigate the possible cellular/circuitry mechanism(s) underlying ultrasound modulation.Main results. It was found that ultrasound stimulation of isolated mouse retina enhanced the background activity of ON-RGCs and OFF-RGCs. In addition, background ultrasound stimulation shortened the light response latency of both ON-RGCs and OFF-RGCs, while enhancing part of the RGCs' (both ON- and OFF-subtypes) light response and decreasing that of the others. In some ON-OFF RGCs, the ON- and OFF-responses of an individual cell were oppositely modulated by the ultrasound stimulation, which suggests that ultrasound stimulation does not necessarily exert its effect directly on RGCs, but rather via its influence on other type(s) of cells. By analyzing the cross-correlation between the firing sequences of RGC pairs, it was found that concerted activity occurred during ultrasound stimulation differed from that occurred during light stimulation, in both spatial and temporal aspects. These results suggest that the cellular circuits involved in ultrasound- and light-induced concerted activities are different and glial cells may be involved in the circuit in response to ultrasound.Significance. These findings demonstrate that ultrasound affects neuronal background activity and light responsiveness, which are critical for visual information processing. These results may also imply a hitherto unrecognized role of glial cell activation in the bidirectional modulation effects of RGCs and may be critical for the nervous system.

Toward the sustainable use of mineral phosphorus fertilizers for crop production in China: From primary resource demand to final agricultural use.

Sustainable phosphorus (P) management is crucial to both food security and environmental conservation. The optimization of P input from mineral fertilizers has been advocated as an effective approach to improve P use efficiency. However, strategies for maximizing P use efficiency by linking soil-crop systems and fertilizer types with the P flow, from a whole P supply chain perspective, are lacking. In this study, a meta-analysis and substance flow analysis (SFA) were employed to evaluate the effects of different mineral P fertilizer types on crop yield and P flow from rock phosphate (RP) exploitation to P use in China. Compared to single superphosphate (SSP), triple superphosphate (TSP), and calcium magnesium phosphate (CMP), a significantly higher yield was obtained when diammonium phosphate (DAP) and monoammonium phosphate (MAP) were used 2005 onwards. However, P loss, from RP extraction to application, was 24% higher for DAP and MAP than for SSP, TSP, and CMP. DAP and MAP use led to a 6% larger P footprint than SSP, TSP, and CMP use. The P use efficiency could be improved by 22%, 36%, and 40% in wheat, maize, and rice production, respectively, by integrating the soil-crop system with mineral P fertilizer types, while P loss and P footprint could be reduced by 13% and 17%, respectively. These results indicate that P use efficiency can be significantly improved by integrating mineral P fertilizer types with soil-crop systems, providing an effective approach for RP exploitation to improve P use efficiency and alleviate the overexploitation of RP.

Spatial and temporal correlations of spike trains in frog retinal ganglion cells.

For a neuron, firing activity can be in synchrony with that of others, which results in spatial correlation; on the other hand, spike events within each individual spike train may also correlate with each other, which results in temporal correlation. In order to investigate the relationship between these two phenomena, population neurons' activities of frog retinal ganglion cells in response to binary pseudo-random checker-board flickering were recorded via a multi-electrode recording system. The spatial correlation index (SCI) and temporal correlation index (TCI) were calculated for the investigated neurons. Statistical results showed that, for a single neuron, the SCI and TCI values were highly related--a neuron with a high SCI value generally had a high TCI value, and these two indices were both associated with burst activities in spike train of the investigated neuron. These results may suggest that spatial and temporal correlations of single neuron's spiking activities could be mutually modulated; and that burst activities could play a role in the modulation. We also applied models to test the contribution of spatial and temporal correlations for visual information processing. We show that a model considering spatial and temporal correlations could predict spikes more accurately than a model does not include any correlation.

Phosphorus-Use Efficiency Modified by Complementary Effects of P Supply Intensity With Limited Root Growth Space.

Space availability and the maintenance of adequate phosphorus (P) supply in the root zone are essential for achieving high yield and P-use efficiency in maize production by manipulating the root morphology and arbuscular mycorrhizal (AM) fungi colonization. A major trade-off exists between root growth and AM colonization that is influenced by soil P supply intensity and space availability. However, how soil P manipulates the root morphological characteristics and AM colonization to compensate for the limitation of root-growth space induced by high-planting density is not clear. Therefore, pot experiments were conducted to investigate interactions between the root growth and AM fungi by optimizing soil P supply to compensate for limited root growth space induced by high-planting density. Similar shoot biomass and P uptake values were obtained in P200 (200 mg P kg-1 soil) under D = 40 (i.e., diameter of the pot is 40 cm) and P400 under D = 30, and similar values were obtained for root length, tap root length, root angle, lateral root density, and AM colonization. However, the improvement in P supply in the root zone, shoot biomass, and P uptake in P400 under D = 20 were lower than in P200 under D = 30, and there were no significant differences in the root parameters between P200 and P400 under D = 20; similarly, the root growth and AM colonization exhibited similar trends. These results suggest that optimizing P supply in the root zone to regulate the interaction between root morphological traits and AM colonization can compensate for limited root-growth space. Although P supply in the root zone increased after the root-growth space was compressed, it could not meet the P demand of maize; thus, to achieve the most efficient use of P under intensive high-density maize production, it is necessary to optimally coordinate root growth space and P supply in the root zone.