Strategies to reduce nutrient pollution from manure management in China

As the demand for livestock products continues to increase in China, so too does the challenge of managing increasing quantities of manure. Urgent action is needed to control point source (housing, storage and processing) and diffuse (field application) pollution and improve the utilization of manure nutrients and organic matter. Here, we review strategies to improve management at each stage of the manure management chain and at different scales. Many strategies require infrastructure investment, e.g., for containment of all manure fractions. Engineering solutions are needed to develop advanced composting systems with lower environmental footprints and design more efficient nutrient stripping technologies. At the field-scale, there is an urgent need to develop a manure nutrient recommendation system that accounts for the range of manure types, cropping systems, soils and climates throughout China. At the regional scale, coordinated planning is necessary to promote recoupling of livestock and cropping systems, and reduce nutrient accumulation in regions with little available landbank, while minimizing the risk of pollution swapping from one region to another. A range of stakeholders are needed to support the step change and innovation required to improve manure management, reduce reliance on inorganic fertilizers, and generate new business opportunities

Interactions between Melanin Enzymes and Their Atypical Recruitment to the Secretory Pathway by Palmitoylation

Melanins are biopolymers that confer coloration and protection to the host organism against biotic or abiotic insults. The level of protection offered by melanin depends on its biosynthesis and its subcellular localization. Previously, we discovered that Aspergillus fumigatus compartmentalizes melanization in endosomes by recruiting all melanin enzymes to the secretory pathway. Surprisingly, although two laccases involved in the late steps of melanization are conventional secretory proteins, the four enzymes involved in the early steps of melanization lack a signal peptide or a transmembrane domain and are thus considered “atypical” secretory proteins. In this work, we found interactions among melanin enzymes and all melanin enzymes formed protein complexes. Surprisingly, the formation of protein complexes by melanin enzymes was not critical for their trafficking to the endosomal system. By palmitoylation profiling and biochemical analyses, we discovered that all four early melanin enzymes were strongly palmitoylated during conidiation. However, only the polyketide synthase (PKS) Alb1 was strongly palmitoylated during both vegetative hyphal growth and conidiation when constitutively expressed alone. This posttranslational lipid modification correlates the endosomal localization of all early melanin enzymes. Intriguingly, bioinformatic analyses predict that palmitoylation is a common mechanism for potential membrane association of polyketide synthases (PKSs) and nonribosomal peptide synthetases (NRPSs) in A. fumigatus. Our findings indicate that protein-protein interactions facilitate melanization by metabolic channeling, while posttranslational lipid modifications help recruit the atypical enzymes to the secretory pathway, which is critical for compartmentalization of secondary metabolism

Secreted Acb1 Contributes to the Yeast-to-Hypha Transition in Cryptococcus neoformans

Adaptation to stress by eukaryotic pathogens is often accompanied by a transition in cellular morphology. The human fungal pathogen Cryptococcus neoformans is known to switch between the yeast and the filamentous form in response to amoebic predation or during mating. As in the classic dimorphic fungal pathogens, the morphotype is associated with the ability of cryptococci to infect various hosts. Many cryptococcal factors and environmental stimuli, including pheromones (small peptides) and nutrient limitation, are known to induce the yeast-to-hypha transition. We recently discovered that secreted matricellular proteins could also act as intercellular signals to promote the yeast-to-hypha transition. Here we show that the secreted acyl coenzyme A (acyl-CoA)-binding protein Acb1 plays an important role in enhancing this morphotype transition. Acb1 does not possess a signal peptide. Its extracellular secretion and, consequently, its function in filamentation are dependent on an unconventional GRASP (Golgi reassembly stacking protein)-dependent secretion pathway. Surprisingly, intracellular recruitment of Acb1 to the secretory vesicles is independent of Grasp. In addition to Acb1, Grasp possibly controls the secretion of other cargos, because the graspΔ mutant, but not the acb1Δ mutant, is defective in capsule production and macrophage phagocytosis. Nonetheless, Acb1 is likely the major or the sole effector of Grasp in terms of filamentation. Furthermore, we found that the key residue of Acb1 for acyl binding, Y80, is critical for the proper subcellular localization and secretion of Acb1 and for cryptococcal morphogenesis

Dermaseptin-PH: A Novel Peptide with Antimicrobial and Anticancer Activities from the Skin Secretion of the South American Orange-Legged Leaf Frog, Pithecopus (Phyllomedusa) hypochondrialis

The dermaseptin peptides, mainly derived from the skin secretions of Hylidae frogs, belong to a superfamily of antimicrobial peptides and exhibit diverse antimicrobial and anticancer activities with low cytotoxicity. Here, we reported a novel dermaseptin peptide, from the South American orange-legged leaf frogs, Pithecopus (Phyllomedusa) hypochondrialis, processing the shortest peptide length, namely Dermaseptin-PH. The complementary DNA (cDNA) encoding biosynthetic precursor of Dermaseptin-PH was initially identified by the rapid amplification of cDNA ends PCR (RACE-PCR) technique from the skin secretion. The predicted primary structure was confirmed by a combination of reverse-phase high performance liquid chromatography (RP-HPLC) and MS/MS fragmentation from the skin secretion. Chemically-synthetic Dermaseptin-PH was investigated using a range of bioactivity assessment assays to evaluate the biological activities and cytotoxicity of Dermaseptin-PH. Dermaseptin-PH inhibited the growth of Gram-negative bacteria, Gram-positive bacteria, and pathogenic yeast Candida albicans. In addition, Dermaseptin-PH showed a broad-spectrum of anticancer activities against several cancer cell lines including MCF-7, H157, U251MG, MDA-MB-435S, and PC-3. The potent antimicrobial and anticancer activities of Dermaseptin-PH make it a promising candidate in the discovery of new drugs for clinical applications, and the relatively short sequence of Dermaseptin-PH can provide new insight for the research and structural modification of new peptide drugs

Long-Term Outcomes of Three-Dimensional High-Dose-Rate Brachytherapy for Locally Recurrent Early T-Stage Nasopharyngeal Carcinoma

Background: Brachytherapy (BT) is one of the techniques available for retreatment of patients with locally recurrent nasopharyng eal carcinoma (rNPC). In this study, we evaluated the treatment outcome and late toxicities of three-dimensional high-dose-rate brachytherapy (3D-HDR-BT) for patients with locally rNPC.Materials and Methods: This is a retrospective study involving 36 patients with histologically confirmed rNPC from 2004 to 2011. Of the 36 patients, 17 underwent combined-modality treatment (CMT) consisting of external beam radiotherapy (EBRT) followed by 3D-HDR-BT, while the other 19 underwent 3D-HDR-BT alone. The median dose of EBRT for the CMT group was 60 (range, 50–66) Gy, with an additional median dose of BT of 16 (range, 9–20) Gy. The median dose for the 3D-HDR-BT group was 32 (range, 20–36) Gy. The measured treatment outcomes were the 5- and 10-year locoregional recurrence-free survival (LRFS), disease-free survival (DFS), overall survival (OS), and late toxicities.Results: The median age at recurrence was 44.5 years. The median follow-up period was 70 (range, 6–142) months. The 5-year LRFS, DFS, and OS for the entire patient group were 75.4, 55.6, and 74.3%, respectively, while the 10-year LRFS, DFS, and OS for the entire patient group were 75.4, 44.2, and 53.7%, respectively. The 10-year LRFS in the CMT group was higher than that in the 3D-HDR-BT-alone group (93.8 vs. 58.8%, HR: 7.595, 95%CI: 1.233–61.826, p = 0.025). No grade 4 late radiotherapy-induced toxicities were observed.Conclusions: 3D-HDR-BT achieves favorable clinical outcomes with mild late toxicity in patients with locally rNPC

Dynamic and Functional Characteristics of Predominant Species in Industrial Paocai as Revealed by Combined DGGE and Metagenomic Sequencing

The microbial community during the fermentation of industrial paocai, a lactic acid fermented vegetable food, was investigated via combined denaturing gradient gel electrophoresis (DGGE) and metagenomic sequencing. Firmicutes and Proteobacteria were identified as the dominant phyla during the fermentation. DGGE results of the bacterial community analysis showed that many genera were observed during the fermentation of industrial paocai, but the same predominant genus and species were observed: Lactobacillus and Lactobacillus (L.) alimentarius/L. paralimentarius. The abundance of L. alimentarius/L. paralimentarius increased fast during the initial stage of fermentation and approximately remained constant during the later stage. Metagenomic sequencing was used to finally identify the predominant species and their genetic functions. Metabolism was the primary functions of the microbial community in industrial paocai fermentation, including carbohydrate metabolism (CM), overview (OV), amino acid metabolism (AAM), nucleotide metabolism (NM), energy metabolism (EM), etc. The predominant species L. alimentarius and L. paralimentarius were involved in plenty of pathways in metabolism and played different roles in the metabolism of carbohydrate, amino acid, lipid to form flavor compounds during industrial paocai fermentation. This study provided valuable information about the predominant species in industrial paocai and its functional properties, which could enable us to advance our understanding of the fermentation mechanism during fermentation of industrial paocai. Our results will advance the understanding of the microbial roles in the industrial paocai fermentation and provide a theoretical basis for improving the quality of industrial paocai products