Characteristics of Biohydrogen Production and Performance of Hydrogen-Producing Acetogen by Increasing Normal Molasses Wastewater Proportion in Anaerobic Baffled Reactor.

The biohydrogen production efficiency and performance of hydrogen-producing acetogen in a four-compartment anaerobic baffled reactor (ABR) were studied by gradually increasing the influent normal molasses wastewater (NMWW) proportion. When the influent NMWW proportion increased to 55%, ABR could develop microbial community with methanogenic function in 63 days and reach a stable operation. When the influent NMWW proportion increased to 80% and reached a stable state, ethanol fermentation was established from butyric acid fermentation in the first three compartments, whereas butyric acid fermentation in the fourth compartment was strengthened. The average biohydrogen production yield and biohydrogen production capacity by COD removal increased to as high as 12.85 L/day and 360.22 L/kg COD when the influent NMWW proportion increased from 55% to 80%, respectively. Although the biogas yield and the specific biogas production rate reached 61.54 L/day and 232 L/kg MLVSS·day, the biohydrogen production yield and specific biohydrogen production rate were only 12.85 L/day and 48 L/kg MLVSS·day, which results in hydrogen consumption by homoacetogenesis and methanogenesis.

Decoding the biochemical dialogue: metabolomic insights into soybean defense strategies against diverse pathogens.

Soybean, a crucial global leguminous crop, confronts persistent threats from diverse pathogens, exerting a profound impact on global yields. While genetic dimensions of soybean-pathogen interactions have garnered attention, the intricate biochemical responses remain poorly elucidated. In this study, we applied targeted and untargeted liquid chromatography coupled to mass spectrometry (LC-MS) metabolite profiling to dissect the complex interplay between soybeans and five distinct pathogens. Our analysis uncovered 627 idMS/MS spectra, leading to the identification of four main modules, encompassing flavonoids, isoflavonoids, triterpenoids, and amino acids and peptides, alongside other compounds such as phenolics. Profound shifts were observed in both primary and secondary metabolism in response to pathogenic infections. Particularly notable were the bidirectional changes in total flavonoids across diverse pathogenic inoculations, while triterpenoids exhibited a general declining trend. Noteworthy among the highly inducible total flavonoids were known representative anti-pathogen compounds (glyceollin I), backbone forms of isoflavonoids (daidzein, genistein, glycitein, formononetin), and newly purified compounds in this study (prunin). Subsequently, we delved into the biological roles of these five compounds, validating their diverse functions against pathogens: prunin significantly inhibited the vegetative growth and virulence of Phytophthora sojae; genistein exhibited a pronounced inhibitory effect on the vegetative growth and virulence of Phomopsis longicolla; daidzein and formononetin displayed significant repressive effects on the virulence of P. longicolla. This study underscores the potent utility of metabolomic tools, providing in-depth insights into plant-pathogen interactions from a biochemical perspective. The findings not only contribute to plant pathology but also offer strategic pathways for bolstering plant resistance against diseases on a broader scale.

Mining oomycete proteomes for phosphatome leads to the identification of specific expanded phosphatases in oomycetes.

Phosphatases are important regulators of protein phosphorylation and various cellular processes, and they serve as counterparts to kinases. In this study, our comprehensive analysis of oomycete complete proteomes unveiled the presence of approximately 3833 phosphatases, with most species estimated to have between 100 and 300 putative phosphatases. Further investigation of these phosphatases revealed a significant increase in protein serine/threonine phosphatases (PSP) within oomycetes. In particular, we extensively studied the metallo-dependent protein phosphatase (PPM) within the PSP family in the model oomycete Phytophthora sojae. Our results showed notable differences in the expression patterns of PPMs throughout 10 life stages of P. sojae, indicating their vital roles in various stages of oomycete pathogens. Moreover, we identified 29 PPMs in P. sojae, and eight of them possessed accessory domains in addition to phosphate domains. We investigated the biological function of one PPM protein with an extra PH domain (PPM1); this protein exhibited high expression levels in both asexual developmental and infectious stages. Our analysis confirmed that PPM1 is indeed an active protein phosphatase, and its accessory domain does not affect its phosphatase activity. To delve further into its function, we generated knockout mutants of PPM1 and validated its essential roles in mycelial growth, sporangia and oospore production, as well as infectious stages. To the best of our knowledge, this study provides the first comprehensive inventory of phosphatases in oomycetes and identifies an important phosphatase within the expanded serine/threonine phosphatase group in oomycetes.

Enhanced proteins and amino acids production based on ammonia nitrogen assimilation and sludge increment by the integration of bioadsorption with anaerobic-anoxic-oxic (AAO) process.

Poor effect of contaminants removal efficiency and low organic matter content of activated sludge are common in wastewater treatment plants (WWTPs) in China due to the low-strength wastewater. An anaerobic-anoxic-oxic (AAO) and an adsorption/AAO (A/AAO) combined system were established simultaneously to conduct a comparative study for realizing the conversion of carbon source in influent and the enrichment and recovery of proteins and amino acids through the assimilation of ammonia nitrogen. The experimental results showed that 63.5% of the organic matter in influent was adsorbed and flocculated in adsorption process, and the removal rates of chemical oxygen demand, total nitrogen and total phosphorus in A/AAO process were 88.7%, 77.1%, and 93.0% respectively, which were remarkably better than those in AAO process owing to the addition of improved carbon source. Ammonia assimilation rate of A/AAO process was 26.7% higher than that of AAO process, which implied that the ammonia used to synthesize sludge protein was prominently increased. Furthermore, intracellular proteins and amino acids in A/AAO process were 20% higher than those of AAO process, and the quality was equivalent with fish meal or soybean meal as feed. In addition, the microbial community analysis based on 16S rDNA was conducted. Dechloromonas, Zoogloea, Nitrospira, and Flavobacterium were the main genera, and played important roles in nutrient removal and ammonia nitrogen assimilation. The integration of adsorption process was significant to low-strength wastewater treatment and the improvement of excess sludge quality, which is a prospective inspiration for the resource recovery-based wastewater treatment process.

Full-scale membrane bioreactor process WWTPs in East Taihu basin: Wastewater characteristics, energy consumption and sustainability.

Based on the collection and analysis of essential data from wastewater treatment plants (WWTPs) in recent ten years, the work provides the wastewater characteristics and energy consumption performance in full-scale membrane bioreactor (MBR) process in East Taihu basin, China. High-quality effluent was achieved although the influent carbon source was not beneficial to total nitrogen and total phosphorus removal. The average specific energy consumption (SEC) was 0.52 kWh/m3, which was remarkably lower than that of full-scale MBR process in developed countries, however, the average SEC value was higher than that of conventional activated sludge (CAS) process in China. In addition, the SEC value was largely reduced in 2018, and the regulation of suction pump and aeration mode were considered as the main control methods. Energy consumption will increase along with the influent volume, while the elevation of COD and NH4+-N reduction will bring about relatively low energy consumption. Furthermore, sustainability index was established to comprehensively evaluate the performance of full-scale MBR process, meaning that with relatively low permeate ratio of effluent, full-scale MBR process presented to be inferior to CAS process in sustainability and not feasible to be applied in the upgradation and construction of WWTPs.

Preclinical Evaluation of a Novel 99mTc-Labeled CB86 for Rheumatoid Arthritis Imaging.

Early diagnosis and therapy are crucial to control disease progression optimally and achieve a good prognosis in rheumatoid arthritis (RA). Previous study showed that a technetium-99m (99mTc)-labeled TSPO ligand (99mTc-CB256 [2-(8-(2-(bis(pyridin-2-yl)methyl)amino)acetamido)-2-(4-chlorophenyl)H-imidazo[1,2-a]pyridin-3-yl)-N,N-dipropylacetamide] composed of a translocator protein (TSPO) ligand CB86 [[2-(4-chlorophenyl)-8-amino-imidazo[1,2-a]-pyridin-3-yl]-N,N-di-n-propylacetamide] and di-(2-picolyl)amine, a bifunctional chelate agent, was used to image a TSPO-rich cancer cell in vitro; however, few 99mTc-CB256 in vivo evaluation has been reported so far probably due to the cytotoxicity of CB256 (ca. 75 times more than analogous CB86). Herein, we describe a novel TSPO targeting radiopharmaceutical consisting of CB86 and diethylenetriaminepentaacetic acid (DTPA), a conventional bifunctional chelating ligand in clinical trials used to prepare 99mTc-labeled CB86, and its evaluation as a 99mTc-single-photon emission computed tomography (SPECT) probe. The radiosynthesis and characterization of 99mTc-DPTA-CB86 including hydrophilicity and stability tests were determined. Additionally, the binding affinity and specificity of 99mTc-DTPA-CB86 to TSPO were evaluated using RAW264.7 macrophage cells. Biodistribution and 99mTc-SPECT studies were conducted on rheumatoid arthritis (RA) rat models after the injection of 99mTc-DTPA-CB86 with or without co-injection of unlabeled DTPA-CB86. The radiosynthesis of 99mTc-DTPA-CB86 was completed successfully with the labeling yields and radiochemical purity of 95.86 ± 2.45 and 97.45 ± 0.69%, respectively. The probe displayed good stability in vitro and binding specificity to RAW264.7 macrophage cells. In the biodistribution studies, 99mTc-DTPA-CB86 exhibited rapid inflammatory ankle accumulation. At 180 min after administration, 99mTc-DTPA-CB86 uptakes of the left inflammatory ankle were 2.35 ± 0.10 percentage of the injected radioactivity per gram of tissue (% ID/g), significantly higher than those of the normal tissues. 99mTc-SPECT imaging studies revealed that 99mTc-DTPA-CB86 could clearly identify the left inflammatory ankle with good contrast at 30-180 min after injection. Therefore, 99mTc-DTPA-CB86 may be a promising probe for arthritis 99mTc-SPECT imaging.

Distribution and diversity of filamentous bacteria in wastewater treatment plants exhibiting foaming of Taihu Lake Basin, China.

Foaming caused by filamentous bacteria in activated sludge (AS) is a common phenomenon in municipal wastewater treatment plants (WWTPs) in Taihu Lake Basin of South China. In this study, total bacterial and filamentous bacterial communities were comprehensively characterized in AS and foams from eight municipal WWTPs by high-throughput sequencing technology. Results showed that alpha diversities of total bacterial communities in foams were obviously lower than those in AS samples. The bacterial community structures were significantly different between WWTPs rather than sample types (AS vs. foam). For most WWTPs, the Actinobacteria phylum was highly enriched in foams and the most abundant genera in foams were common mycolata. Sixteen filamentous bacteria were identified against the improved bulking and foaming bacteria (BFB) database. Abundance and composition of BFB in different WWTPs and different sample types were significantly different. 'Nostocoida limicola' I Trichococcus and Microthrix were generally dominant in AS samples. The dominant BFB in foams were associated with Microthrix, Skermania, Gordonia, and Mycobacterium. A new Defluviicoccus spp. in cluster III was identified in severe and continuous foams. Moreover, dominant BFB in stable and continuous foams with light level in one typical WWTP were diverse, even, and dynamic. Bacterial co-occurrence network analysis implied that the bacterial community of AS was more sensitive to disturbance than that of foam.

The use of Pseudomonas fluorescens P13 to control sclerotinia stem rot (Sclerotinia sclerotiorum) of oilseed rape.

Sclerotinia stem rot (SSR) caused by the fungus Sclerotinia sclerotiorum has been an increasing threat to oilseed rape (Brassica napus L.) cultivation. Efficient and environment-friendly treatments are much needed. Here we focus on microbial control. The Pseudomonas fluorescens P13 that was isolated from oilseed rape cultivation soil, proved to be a useful biocontrol strain for application. Morphology, physiological and biochemical tests and 16S rDNA analysis demonstrated that it was P. fluorescens P13 and that it had a broad antagonistic spectrum, significantly lessening the mycelial growth of S. sclerotiorum by 84.4% and suppressing sclerotial formation by 95-100%. Scanning electron microscopy studies attested that P13 deformed S. sclerotiorum mycelia when they were cultured together. P13 did not produce chitinase but did produce hydrogen cyanide (HCN) which was likely one of the antagonistic mechanisms. The density of P13 remained at a high level (≥10(6) CFU/ml) during 5 weeks in the rhizosphere soil and roots. P13 reduced SSR severity at least by 59% in field studies and also promoted seedling growth (p<0.05) at the seedling stage. From these data, our work provided evidence that P13 could be a good alternative biological resource for biocontrol of S. sclerotiorum.

Characterization of the N-terminal domain of classical swine fever virus RNA-dependent RNA polymerase.

To investigate RNA-dependent RNA polymerase (RdRp) further, mutational analysis of the N-terminal domain of the NS5B protein of Classical swine fever virus was performed. Results show that the N-terminal domain (positions 1-300) of the protein might be divided artificially into four different regions, N1-N4. The N1 region (positions 1-61) contained neither conserved lysine nor conserved arginine residues. NS5B protein with deletion of the N1 region has the capacity for elongative RNA synthesis, but not for de novo RNA synthesis on natural templates. All substitutions of the conserved lysines and arginines in the N2 region (positions 63-216) destroyed RdRp activity completely. Substitutions of the conserved arginines in the N3 region (positions 217-280) seriously reduced RdRp activity. However, all substitutions of the conserved lysines in this region enhanced RNA synthesis and made the mutants synthesize RNA on any template. Substitutions of the conserved arginines in the N4 region (positions 281-300) reduced elongative synthesis and destroyed de novo RNA synthesis. In contrast, substitutions of lysines in this region did not affect RdRp activity significantly. These data indicate that the N3 region might be related to the enzymic specificity for templates, and the conserved lysines and arginines in different regions have different effects on RdRp activity. In combination with the published crystal structure of bovine viral diarrhea virus NS5B, these results define the important role of the N-terminal domain of NS5B for template recognition and de novo RNA synthesis.