Pyroptosis-mediator GSDMD promotes Parkinson's disease pathology via microglial activation and dopaminergic neuronal death.

GSDMD-mediated pyroptosis occurs in the nigrostriatal pathway in Parkinson's disease animals, yet the role of GSDMD in neuroinflammation and death of dopaminergic neurons in Parkinson's disease remains elusive. Here, our in vivo and in vitro studies demonstrated that GSDMD, as a pyroptosis executor, contributed to glial reaction and death of dopaminergic neurons across different Parkinson's disease models. The ablation of the Gsdmd attenuated Parkinson's disease damage by reducing dopaminergic neuronal death, microglial activation, and detrimental transformation. Disulfiram, an inhibitor blocking GSDMD pore formation, efficiently curtailed pyroptosis, thereby lessening the pathology of Parkinson's disease. Additionally, a modification in GSDMD was identified in the blood of Parkinson's disease patients in contrast to healthy subjects. Therefore, the detected alteration in GSDMD within the blood of Parkinson's disease patients and the protective impact of disulfiram could be promising for the diagnostic and therapeutic approaches against Parkinson's disease.

Hypoxia inducible factor-1α regulates microglial innate immune memory and the pathology of Parkinson's disease.

Neuroinflammation is one of the core pathological features of Parkinson's disease (PD). Innate immune cells play a crucial role in the progression of PD. Microglia, the major innate immune cells in the brain, exhibit innate immune memory effects and are recognized as key regulators of neuroinflammatory responses. Persistent modifications of microglia provoked by the first stimuli are pivotal for innate immune memory, resulting in an enhanced or suppressed immune response to second stimuli, which is known as innate immune training and innate immune tolerance, respectively. In this study, LPS was used to establish in vitro and in vivo models of innate immune memory. Microglia-specific Hif-1α knockout mice were further employed to elucidate the regulatory role of HIF-1α in innate immune memory and MPTP-induced PD pathology. Our results showed that different paradigms of LPS could induce innate immune training or tolerance in the nigrostriatal pathway of mice. We found that innate immune tolerance lasting for one month protected the dopaminergic system in PD mice, whereas the effect of innate immune training was limited. Deficiency of HIF-1α in microglia impeded the formation of innate immune memory and exerted protective effects in MPTP-intoxicated mice by suppressing neuroinflammation. Therefore, HIF-1α is essential for microglial innate immune memory and can promote neuroinflammation associated with PD.

Uncovering interactions among ternary electron donors of organic carbon source, thiosulfate and Fe0 in mixotrophic advanced denitrification: Proof of concept from simulated to authentic secondary effluent.

To offset the imperfections of higher cost and emission of CO2 greenhouse gas in heterotrophic denitrification (HDN) as well as longer start-up time in autotrophic denitrification (ADN), we synergized the potential ternary electron donors of organic carbon source, thiosulfate and zero-valent iron (Fe0) to achieve efficient mixotrophic denitrification (MDN) of oligotrophic secondary effluent. When the influent chemical oxygen demand to nitrogen (COD/N) ratio ascended gradually in the batch operation with sufficient sulfur to nitrogen (S/N) ratio, the MDN with thiosulfate and Fe0 added achieved the highest TN removal for treating simulated and authentic secondary effluents. The external carbon is imperative for initiating MDN, while thiosulfate is indispensable for promoting TN removal efficiency. Although Fe0 hardly donated electrons for denitrification, the suitable circumneutral environment for denitrification was implemented by OH- released from Fe0 corrosion, which neutralized H+generated during thiosulfate-driven ADN. Meanwhile, Fe0 corrosion consumed the dissolved oxygen (DO) and created the low DO environment suitable for anoxic denitrification. This process was further confirmed by the continuous flow operation for treating authentic secondary effluent. The TN removal efficiency achieved its maximum under the combination condition of influent COD/N ratio of 3.1-3.5 and S/N ratio of 2.0-2.1. Whether in batch or continuous flow operation, the coordination of thiosulfate and Fe0 maintained the dominance of Thiobacillus for ADN, with the dominant heterotrophic denitrifiers (e.g., Plasticicumulans, Terrimonas, Rhodanobacter and KD4-96) coexisting in MDN system. The interaction insights of ternary electron donors in MDN established a pathway for realizing high-efficiency nitrogen removal of secondary effluent.

TLR2 deficiency is beneficial at the late phase in MPTP-induced Parkinson' disease mice.

AIMS: Parkinson's disease (PD) is a progressive neurodegenerative disorder. The etiology of PD is still elusive but neuroinflammation is proved to be an important contributor. Toll-like receptor 2 (TLR2) involves in the release of several inflammatory cytokines. Whether TLR2 serves as a mediator contributing to the damage of DA system in PD remain unclear. MAIN METHODS: Tlr2 knockout (Tlr2-/-) and wild-type (WT) mice were treated with a subacute regimen of 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). At 3, 7 and 14 days after MPTP injection, the behavioral performance, including the Pole test, the Rotarod test, the Rearing test and the Wire hang test was evaluated. Moreover, the PD-like phenotypes, including dopaminergic degeneration, the activation of glial cells and the α-Syn expression were systematically analyzed in the nigrostriatal pathway. Finally, the composition of gut microbiota in the MPTP-treated groups were assessed. KEY FINDINGS: TLR2 deficiency had no obvious impact on the dopaminergic injury at 3 and 7 days following MPTP administration. On the contrary, at 14 days post injection, TLR2 deficiency not only significantly attenuated motor deficits in the Pole test and the Rotarod test, and the nigrostriatal dopaminergic degeneration, but also mitigated α-Syn abnormality, astrocyte activation and neuroinflammation through the suppressed TLR2/MyD88/TRAF6/NF-κB signaling pathways. Additionally, the alteration of gut microbiota was also detected in the mutant mice. SIGNIFICANCE: These findings highlight the neuroprotective effect of TLR2-pathways at the late phase in the MPTP-induced PD mouse model.

Proline Metabolism Process and Antioxidant Potential of Lycium ruthenicum Murr. in Response to NaCl Treatments.

Salinity influences the level of antioxidants and proline content, which are both involved in the regulation of stress responses in plants. To examine the interplay between the antioxidant system and proline metabolism in plant stress acclimation, explants of Lycium ruthenicum were subjected to NaCl treatments, and the growth characteristics, antioxidant enzyme activities, proline accumulation, and metabolic enzyme content were analyzed. The results revealed that NaCl concentrations between 50 to 150 mM have a positive effect on the growth of L. ruthenicum explants. Increasing NaCl concentrations elevated the activities of superoxide dismutase (SOD) and catalase (CAT), while hydrogen peroxide (H2O2) content was inhibited, suggesting that the elevated antioxidants play a central protective role in superoxide anion (O2•-) and H2O2 scavenging processes in response to NaCl treatments. Also, high proline levels also protect antioxidant enzyme machinery, thus protecting the plants from oxidative damage and enhancing osmotic adjustment. Increasing levels of pyrroline-5-carboxylate synthetase (P5CS), pyrroline-5-carboxylate reductase (P5CR), and ornithine-δ-aminotransferase (δ-OAT) were observed, resulting in elevated level of proline. In addition, the expression levels of LrP5CS1, -2, -3, LrOAT-1, and -2 were promoted in NaCl treatments. According to the combined analysis of metabolic enzyme activities and their relative expression, it is confirmed that the glutamate (Glu) pathway is activated in L. ruthenicum faced with different levels of NaCl concentrations. However, Glu supplied by δ-OAT is fed back into the main pathway for proline metabolism.

HDAC6 Deficiency Has Moderate Effects on Behaviors and Parkinson's Disease Pathology in Mice.

Histone deacetylase 6 (HDAC6) is involved in the regulation of protein aggregation and neuroinflammation, but its role in Parkinson's disease (PD) remains controversial. In this study, Hdac6-/- mice were generated by CRISPR-Cas9 technology for exploring the effect of HDAC6 on the pathological progression of PD. We found that male Hdac6-/- mice exhibit hyperactivity and certain anxiety. In the acute 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP)-induced PD mice, though motor injury was slightly alleviated by HDAC6 deficiency, dopamine (DA) depletion in the striatum, the decrease in the number of DA neurons in the substantia nigra (SN) and the reduction in DA neuronal terminals were not affected. In addition, activation of glial cells and the expression of α-synuclein, as well as the levels of apoptosis-related proteins in the nigrostriatal pathway, were not changed in MPTP-injected wild-type and Hdac6-/- mice. Therefore, HDAC6 deficiency leads to moderate alterations of behaviors and Parkinson's disease pathology in mice.

Comprehensive analysis of microbiota signature across 32 cancer types.

Microbial communities significantly inhabit the human body. Evidence shows the interaction between the human microbiome and host cells plays a central role in multiple physiological processes and organ microenvironments. However, the majority of related studies focus on gut microbiota or specific tissues/organs, and the component signature of intratumor microbiota across various cancer types remains unclear. Here, we systematically analyzed the correlation between intratumor microbial signature with survival outcomes, genomic features, and immune profiles across 32 cancer types based on the public databases of Bacteria in Cancer (BIC) and The Cancer Genome Atlas (TCGA). Results showed the relative abundance of microbial taxa in tumors compared to normal tissues was observed as particularly noticeable. Survival analysis found that specific candidate microbial taxa were correlated with prognosis across various cancers. Then, a microbial-based scoring system (MS), which was composed of 64 candidate prognostic microbes, was established. Further analyses showed significant differences in survival status, genomic function, and immune profiles among the distinct MS subgroups. Taken together, this study reveals the diversity and complexity of microbiomes in tumors. Classifying cancer into different subtypes based on intratumor microbial signatures might reasonably reflect genomic characteristics, immune features, and survival status.

A novel fatty-acid metabolism-based classification for triple negative breast cancer.

BACKGROUND: The high heterogeneity of triple negative breast cancer (TNBC) is the main clinical challenge for individualized therapy. Considering that fatty acid metabolism (FAM) plays an indispensable role in tumorigenesis and development of TNBC, we proposed a novel FAM-based classification to characterize the tumor microenvironment immune profiles and heterogeneous for TNBC. METHODS: Weighted gene correlation network analysis (WGCNA) was performed to identify FAM-related genes from 221 TNBC samples in Molecular Taxonomy of Breast Cancer International Consortium (METABRIC) dataset. Then, non-negative matrix factorization (NMF) clustering analysis was applied to determine FAM clusters based on the prognostic FAM-related genes, which chosen from the univariate/multivariate Cox regression model and the least absolute shrinkage and selection operator (LASSO) regression algorithm. Then, a FAM scoring scheme was constructed to further quantify FAM features of individual TNBC patient based on the prognostic differentially expressed genes (DEGs) between different FAM clusters. Systematically analyses were performed to evaluate the correlation between the FAM scoring system (FS) with survival outcomes, genomic characteristics, tumor microenvironment (TME) features and immunotherapeutic response for TNBC, which were further validated in the Cancer Genome Atlas (TCGA) and GSE58812 datasets. Moreover, the expression level and clinical significancy of the selected FS gene signatures were further validated in our cohort. RESULTS: 1860 FAM-genes were screened out using WGCNA. Three distinct FAM clusters were determined by NMF clustering analysis, which allowed to distinguish different groups of patients with distinct clinical outcomes and tumor microenvironment (TME) features. Then, prognostic gene signatures based on the DEGs between different FAM clusters were identified using univariate Cox regression analysis and Lasso regression algorithm. A FAM scoring scheme was constructed, which could divide TNBC patients into high and low-FS subgroups. Low FS subgroup, characterized by better prognosis and abundance with effective immune infiltration. While patients with higher FS were featured with poorer survival and lack of effective immune infiltration. In addition, two independent immunotherapy cohorts (Imvigor210 and GSE78220) confirmed that patients with lower FS demonstrated significant therapeutic advantages from anti-PD-1/PD-L1 immunotherapy and durable clinical benefits. Further analyses in our cohort found that the differential expression of CXCL13, FBP1 and PLCL2 were significantly associated with clinical outcomes of TNBC samples. CONCLUSIONS: This study revealed FAM plays an indispensable role in formation of TNBC heterogeneity and TME diversity. The novel FAM-based classification could provide a promising prognostic predictor and guide more effective immunotherapy strategies for TNBC.

Deficiency of miR-29a/b1 leads to premature aging and dopaminergic neuroprotection in mice.

Parkinson's disease (PD) is a neurodegenerative disorder characterized by progressive degeneration of midbrain dopaminergic neurons. The miR-29s family, including miR-29a and miR-29b1 as well as miR-29b2 and miR-29c, are implicated in aging, metabolism, neuronal survival, and neurological disorders. In this study, the roles of miR-29a/b1 in aging and PD were investigated. miR-29a/b1 knockout mice (named as 29a KO hereafter) and their wild-type (WT) controls were used to analyze aging-related phenotypes. After challenged with the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), dopaminergic injuries, glial activation, and mouse behaviors were evaluated. Primary glial cells were further cultured to explore the underlying mechanisms. Additionally, the levels of miR-29s in the cerebrospinal fluid (CSF) of PD patients (n = 18) and healthy subjects (n = 17) were quantified. 29a KO mice showed dramatic weight loss, kyphosis, and along with increased and deepened wrinkles in skins, when compared with WT mice. Moreover, both abdominal and brown adipose tissues reduced in 29a KO mice, compared to their WT counterpart. However, in MPTP-induced PD mouse model, the deficiency of miR-29a/b1 led to less severe damages of dopaminergic system and mitigated glial activation in the nigrostriatal pathway, and subsequently alleviated the motor impairments in 3-month-old mice. Eight-month-old mutant mice maintained such a resistance to MPTP intoxication. Mechanistically, the deficiency of miR-29a/b-1 promoted the expression of neurotrophic factors in 1-Methyl-4-phenylpyridinium (MPP+)-treated primary mixed glia and primary astrocytes. In lipopolysaccharide (LPS)-treated primary microglia, knockout of miR-29a/b-1 inhibited the expression of inflammatory factors, and promoted the expression of anti-inflammatory factors and neurotrophic factors. Knockout of miR-29a/b1 increased the activity of AMP-activated protein kinase (AMPK) and repressed NF-κB/p65 signaling in glial cells. Moreover, we found miR-29a level was increased in the CSF of patients with PD. Our results suggest that 29a KO mice display the peripheral premature senility. The combined effects of less activated glial cells might contribute to the mitigated inflammatory responses and elicit resistance to MPTP intoxication in miR-29a/b1 KO mice.

Atmospheric water vapor and soil moisture jointly determine the spatiotemporal variations of CO2 fluxes and evapotranspiration across the Qinghai-Tibetan Plateau grasslands.

Alpine grasslands play important functions in mitigating climate change and regulating water resources. However, the spatiotemporal variability of their carbon and water budgets remains unquantified. Here, 47 site-year observations of CO2 and water vapor fluxes (ET) are analyzed at sites situated along a hydrothermal gradient across the Qinghai-Tibetan Plateau, including an alpine wetland (wettest), an alpine shrub (coldest), an alpine meadow, an alpine meadow-steppe, and an alpine steppe (driest and warmest). The results show that the benchmarks for annual net ecosystem exchange (NEE) are -79.3, -77.8, -66.7, 20.2, and 100.9 g C m-2 year-1 at the meadow, shrub, meadow-steppe, steppe, and wetland, respectively. The peak daily NEE normalized by peak leaf area index converges to 0.93 g C m-2 d-1 at the 5 sites. Except in the wetland (722.8 mm), the benchmarks of annual ET fluctuate from 511.0 mm in the steppe to 589.2 mm in the meadow. Boosted regression trees-based analysis suggests that the enhanced vegetation index (EVI) and net radiation (Rn) determine the variations of growing season monthly CO2 fluxes and ET, respectively, although the effect is to some extent site-specific. Inter-annual variability in NEE, ecosystem respiration (RES), and ET are tightly (R2 > 0.60) related to the inter-growing season NEE, RES, and ET, respectively. Both annual RES and annual NEE are significantly constrained by annual gross primary productivity (GPP), with 85% of the per-unit GPP contributing to RES (R2 = 0.84) and 15% to NEE (R2 = 0.12). Annual GPP significantly correlates with annual ET alone at the drier sites of the meadow-steppe and the steppe, suggesting the coupling of carbon and water is moisture-dependent in alpine grasslands. Over half of the inter-annual spatial variability in GPP, RES, NEE, and ET is explained by EVI, atmospheric water vapor, topsoil water content, and bulk surface resistance (rs), respectively. Because the spatial variations of EVI and rs are strongly regulated by atmospheric water vapor (R2 = 0.48) and topsoil water content (R2 = 0.54), respectively, we conclude that atmospheric water vapor and topsoil water content, rather than the expected air/soil temperatures, drive the spatiotemporal variations in CO2 fluxes and ET across temperature-limited grasslands. These findings are critical for improving predictions of the carbon sequestration and water holding capacity of alpine grasslands.

Declining human activity intensity on alpine grasslands of the Tibetan Plateau.

Climate change and human activities have profoundly changed the structure and functioning of alpine grassland ecosystems on the Tibetan Plateau, the most critical ecological safety shelter for Asia. However, it remains unclear to what degree human activity intensity has impacted the alpine grasslands of the Tibetan Plateau. Here we quantify human activity intensity on alpine grasslands of the Tibetan Plateau based on the relationship between actual and potential net primary production. We found that human activity intensity decreased by 16.1% from 2000 to 2017 across the alpine grasslands, which might be driven by recent ecological conservation policies, especially reductions in livestock numbers. Critical thresholds, which show marked grassland responses to different levels of human disturbances, were identified for each ecozone. The net primary production of dry grasslands on the western ecozones was more resistant to human disturbances but with lower resilience than other alpine grasslands on the plateau. Our findings are beneficial to design practical countermeasures to adapt to climate change and recover damaged grasslands on Tibetan Plateau.

Warming homogenizes apparent temperature sensitivity of ecosystem respiration.

Warming-induced carbon loss through terrestrial ecosystem respiration (Re) is likely getting stronger in high latitudes and cold regions because of the more rapid warming and higher temperature sensitivity of Re (Q 10). However, it is not known whether the spatial relationship between Q 10 and temperature also holds temporally under a future warmer climate. Here, we analyzed apparent Q 10 values derived from multiyear observations at 74 FLUXNET sites spanning diverse climates and biomes. We found warming-induced decline in Q 10 is stronger at colder regions than other locations, which is consistent with a meta-analysis of 54 field warming experiments across the globe. We predict future warming will shrink the global variability of Q 10 values to an average of 1.44 across the globe under a high emission trajectory (RCP 8.5) by the end of the century. Therefore, warming-induced carbon loss may be less than previously assumed because of Q 10 homogenization in a warming world.

Assessment of the vulnerability of alpine grasslands on the Qinghai-Tibetan Plateau.

Assessing ecosystem vulnerability to climate change is critical for sustainable and adaptive ecosystem management. Alpine grasslands on the Qinghai-Tibetan Plateau are considered to be vulnerable to climate change, yet the ecosystem tends to maintain stability by increasing resilience and decreasing sensitivity. To date, the spatial pattern of grassland vulnerability to climate change and the mechanisms that vegetation applies to mitigate the impacts of climate change on grasslands by altering relevant ecosystem characteristics, especially sensitivity and resilience, remain unknown. In this study, we first assessed the spatial pattern of grassland vulnerability to climate change by integrating exposure, sensitivity, and resilience simultaneously, and then identified its driving forces. The results show that grasslands with high vulnerability were mainly located on the edges of the plateau, whereas alpine grasslands in the hinterlands of the plateau showed a low vulnerability. This spatial pattern of alpine grassland vulnerability was controlled by climatic exposure, and grassland sensitivity and resilience to climate change might also exacerbate or alleviate the degree of vulnerability. Climate change had variable impacts on different grassland types. Desert steppes were more vulnerable to climate change than alpine meadows and alpine steppes because of the high variability in environmental factors and their low ability to recover from perturbations. Our findings also confirm that grazing intensity, a quantitative index of the most important human disturbance on alpine grasslands in this plateau, was significantly correlated with ecosystem vulnerability. Moderate grazing intensity was of benefit for increasing grassland resilience and then subsequently reducing grassland vulnerability. Thus, this study suggests that future assessments of ecosystem vulnerability should not ignore anthropogenic disturbances, which might benefit environmental protection and sustainable management of grasslands on the Qinghai-Tibetan Plateau.

SNAP25/syntaxin4/VAMP2/Munc18-1 Complexes in Spinal Dorsal Horn Contributed to Inflammatory Pain.

Soluble N-ethylmaleimide-sensitive factor attachment protein receptors (SNAREs) have been implicated in the trafficking of postsynaptic glutamate receptors, including N-methyl-d-aspartate (NMDA)-subtype glutamate receptors (NMDARs) that are critical for nociceptive plasticity and behavioral sensitization. However, the components of SNAREs complex involved in spinal nociceptive processing remain largely unknown. Here we found that SNAP25, syntaxin4, VAMP2 and Munc18-1 were localized at postsynaptic sites and formed the complex in the superficial lamina of spinal cord dorsal horn of rats. The complex formation between these SNAREs components were accelerated after intraplantar injection of complete Freund's adjuvant (CFA), pharmacological removal of GABAergic inhibition or activation of NMDAR in intact rats. The increased SNAP25/syntaxin4/VAMP2/Munc18-1 interaction facilitated the surface delivery and synaptic accumulation of NMDAR during inflammatory pain. Disruption of the molecular interaction between SNAP25 with its SNARE partners by using a blocking peptide derived from the C-terminus of SNAP25 effectively repressed the surface and synaptic accumulation of GluN2B-containing NMDARs in CFA-injected rats. This peptide also alleviated inflammatory mechanical allodynia and thermal hypersensitivity. These data suggested that SNAREs complex assembly in spinal cord dorsal horn was involved in the inflammatory pain hypersensitivity through promoting NMDAR synaptic trafficking.

Reconsidering the efficiency of grazing exclusion using fences on the Tibetan Plateau.

Grazing exclusion using fences is a key policy being applied by the Chinese government to rehabilitate degraded grasslands on the Tibetan Plateau (TP) and elsewhere. However, there is a limited understanding of the effects of grazing exclusion on alpine ecosystem functions and services and its impacts on herders' livelihoods. Our meta-analyses and questionnaire-based surveys revealed that grazing exclusion with fences was effective in promoting aboveground vegetation growth for up to four years in degraded alpine meadows and for up to eight years in the alpine steppes of the TP. Longer-term fencing did not bring any ecological and economic benefits. We also found that fencing hindered wildlife movement, increased grazing pressure in unfenced areas, lowered the satisfaction of herders, and rendered substantial financial costs to both regional and national governments. We recommend that traditional free grazing should be encouraged if applicable, short-term fencing (for 4-8 years) should be adopted in severely degraded grasslands, and fencing should be avoided in key wildlife habitat areas, especially the protected large mammal species.

Pulmonary embolism as the initial manifestation of right atrial myxoma: A case report and review of the literature.

RATIONALE: Pulmonary embolisms (PEs) are caused by emboli, which mostly originate from deep venous thrombi that travel to and suddenly block the pulmonary arteries. The emboli are usually thrombi, and right atrial myxoma emboli are rare. PATIENT CONCERNS: A 55-year-old man presented with shortness of breath and syncope. We proceeded with computed tomography pulmonary angiography (CTPA) and transthoracic echocardiogram (TTE), the results of which suggested that the diagnosis was a right atrial mass. DIAGNOSIS: A definitive diagnosis compatible with a right atrial myxoma (RAM) with tumoral pulmonary emboli after surgical excision was made. INTERVENTION: Right atrial and pulmonary artery embolectomy. OUTCOMES: The patient followed an uneventful course during the 6 years of follow-up after surgery. According to a review of the literature, RAMs are often not diagnosed in a timely manner or even go completely undiagnosed. TTE, transesophageal echocardiography (TEE), CT, magnetic resonance imaging (MRI), and positron emission tomography/computed tomography may be helpful in the preoperative diagnosis. Surgical removal of the masses from the atrium and pulmonary arteries was relatively uneventful. LESSONS: RAMs should be considered unlikely reasons for fatal pulmonary embolisms.

High Below-Ground Productivity Allocation of Alpine Grasslands on the Northern Tibet.

The allocation of net primary production (NPP) between above- and belowground components is a key step of ecosystem material cycling and energy flows, which determines many critical parameters, e.g., the fraction of below ground NPP (BNPP) to NPP (fBNPP) and root turnover rates (RTR), in vegetation models. However, direct NPP estimation and partition are scarcely based on field measurements of biomass dynamics in the alpine grasslands on the Northern Tibetan Plateau (NTP). Consequently, these parameters are unverifiable and controversial. Here, we measured above- and belowground biomass dynamics (monthly from May to September each year from 2013 to 2015) to estimate NPP dynamics and allocations in four typical alpine grassland ecosystems, i.e., an alpine meadow, alpine meadow steppe, alpine steppe and alpine desert steppe. We found that NPP and its components, above and below ground NPP (ANPP and BNPP), increased significantly from west to east on the NTP, and ANPP was mainly affected by temperature while BNPP and NPP were mainly affected by precipitation. The bulk of BNPP was generally concentrated in the top 10 cm soil layers in all four alpine grasslands (76.1% ± 9.1%, mean ± SD). Our results showed that fBNPP was significantly different among these four alpine grasslands, with its means in alpine meadow (0.93), alpine desert steppe (0.92) being larger than that in the alpine meadow steppe (0.76) and alpine steppe (0.77). Both temperature and precipitation had significant and positive effects on the fBNPP, while their interaction effects were significantly opposite. RTR decreased with increasing precipitation, but increased with increasing temperature across this ecoregion. Our study illustrated that alpine grasslands on the NTP, especially in the alpine meadow and alpine desert steppe, partitioned an unexpected and greater NPP to below ground than most historical reports across global grasslands, indicating a more critical role of the root carbon pool in carbon cycling in alpine grasslands on the NTP.

Design and synthesis of new N-terminal fatty acid modified-antimicrobial peptide analogues with potent in vitro biological activity.

Developing novel antimicrobial agents is a top priority in fighting against bacterial resistance. Thus, a series of new monomer and dimer peptides were designed and synthesized by conjugating fatty acids at the N-terminus of partial d-amino acid substitution analogues of anoplin and dimerization. The new peptides exhibited more efficient killing of gram-negative and gram-positive bacteria, including methicillin-resistant Staphylococcus aureus compared with the parent peptide anoplin, and the dimer peptides were superior to the monomer peptides. It was important that the new peptides displayed low impact on bacterial resistance development. In addition, the antimicrobial activities were not significantly influenced by a physiological salt environment. They also presented high stability in the presence of protease or serum. Almost all of the new peptides had better selectivity towards anionic bacterial membranes over zwitterionic mammalian cell membranes. Moreover, the new peptides displayed synergistic or additive effects when used together with the antibiotics rifampicin and polymyxin B. These results showed that the new peptides could also prevent the formation of bacterial biofilms. Furthermore, outer/inner membrane permeabilization and cytoplasmic membrane depolarization experiments revealed that the new peptides had strong membrane permeabilization and depolarization. Confocal laser scanning microscopy, flow cytometry analysis and scanning electron microscopy further demonstrated that the new peptides could damage the integrity of the bacterial membrane. Finally, a DNA-binding affinity assay showed that the new peptides could bind to bacterial DNA. In summary, the conjugation of fatty acids at the N-terminus of peptides and dimerization are promising strategies for obtaining potent antimicrobial agents.