•A new heterotrophic nitrifying and metal resistant bacterium, named S1, was isolated and identified as Cupriavidus sp.•Strain S1 possessed excellent heterotrophic nitrification and aerobic ...denitrification ability.•It could utilize different forms of nitrogen source in simultaneous nitrification and denitrification.•NH4+ could effectually remove with high concentration heavy metal involved in.
A novel heterotrophic nitrifying and metal resistant bacterium was isolated and identified as Cupriavidus sp. S1. The utilization of ammonium, nitrate and nitrite as well as the production of N2 proved the heterotrophic nitrification and aerobic denitrification ability of S1. The ammonium, nitrate and nitrite removal efficiencies were 99.68%, 98.03% and 99.81%, with removal rates of 10.43, 8.64 and 8.36mg/L/h, respectively. A multiple regression equation well described the relationship between carbon source utilization, cell growth and nitrification. Keeping the shaking speed at 120rpm was beneficial for denitrification. Moreover, different forms of nitrogen source could be utilize in simultaneous nitrification and denitrification. Additionally, the efficient removal of ammonium occurred at 20.0mg/LZn2+, or 10.0mg/LNi2+ or 8.0mg/LCu2+ or 5.0mg/LCr6+, 33.35mmol/L sodium pyruvate, C/N 12–28. These findings demonstrate that S1 was effective for nitrogen removal in industrial wastewater containing heavy metal.
Bisulfite sequencing detects 5mC and 5hmC at single-base resolution. However, bisulfite treatment damages DNA, which results in fragmentation, DNA loss, and biased sequencing data. To overcome these ...problems, enzymatic methyl-seq (EM-seq) was developed. This method detects 5mC and 5hmC using two sets of enzymatic reactions. In the first reaction, TET2 and T4-BGT convert 5mC and 5hmC into products that cannot be deaminated by APOBEC3A. In the second reaction, APOBEC3A deaminates unmodified cytosines by converting them to uracils. Therefore, these three enzymes enable the identification of 5mC and 5hmC. EM-seq libraries were compared with bisulfite-converted DNA, and each library type was ligated to Illumina adaptors before conversion. Libraries were made using NA12878 genomic DNA, cell-free DNA, and FFPE DNA over a range of DNA inputs. The 5mC and 5hmC detected in EM-seq libraries were similar to those of bisulfite libraries. However, libraries made using EM-seq outperformed bisulfite-converted libraries in all specific measures examined (coverage, duplication, sensitivity, etc.). EM-seq libraries displayed even GC distribution, better correlations across DNA inputs, increased numbers of CpGs within genomic features, and accuracy of cytosine methylation calls. EM-seq was effective using as little as 100 pg of DNA, and these libraries maintained the described advantages over bisulfite sequencing. EM-seq library construction, using challenging samples and lower DNA inputs, opens new avenues for research and clinical applications.
High-throughput sequencing (HTS) has become a powerful tool for the detection of and sequence characterization of microRNAs (miRNA) and other small RNAs (sRNA). Unfortunately, the use of HTS data to ...determine the relative quantity of different miRNAs in a sample has been shown to be inconsistent with quantitative PCR and Northern Blot results. Several recent studies have concluded that the major contributor to this inconsistency is bias introduced during the construction of sRNA libraries for HTS and that the bias is primarily derived from the adaptor ligation steps, specifically where single stranded adaptors are sequentially ligated to the 3' and 5'-end of sRNAs using T4 RNA ligases. In this study we investigated the effects of ligation bias by using a pool of randomized ligation substrates, defined mixtures of miRNA sequences and several combinations of adaptors in HTS library construction. We show that like the 3' adaptor ligation step, the 5' adaptor ligation is also biased, not because of primary sequence, but instead due to secondary structures of the two ligation substrates. We find that multiple secondary structural factors influence final representation in HTS results. Our results provide insight about the nature of ligation bias and allowed us to design adaptors that reduce ligation bias and produce HTS results that more accurately reflect the actual concentrations of miRNAs in the defined starting material.
Photoelectrochemical water splitting has long been considered an ideal approach to producing green hydrogen by utilizing solar energy. However, the limited photocurrents and large overpotentials of ...the anodes seriously impede large-scale application of this technology. Here, we use an interfacial engineering strategy to construct a nanostructural photoelectrochemical catalyst by incorporating a semiconductor CdS/CdSe-MoS
and NiFe layered double hydroxide for the oxygen evolution reaction. Impressively, the as-prepared photoelectrode requires an low potential of 1.001 V vs. reversible hydrogen electrode for a photocurrent density of 10 mA cm
, and this is 228 mV lower than the theoretical water splitting potential (1.229 vs. reversible hydrogen electrode). Additionally, the generated current density (15 mA cm
) of the photoelectrode at a given overpotential of 0.2 V remains at 95% after long-term testing (100 h). Operando X-ray absorption spectroscopy revealed that the formation of highly oxidized Ni species under illumination provides large photocurrent gains. This finding opens an avenue for designing high-efficiency photoelectrochemical catalysts for successive water splitting.
Accurately detecting distant evolutionary relationships between proteins remains an ongoing challenge in bioinformatics. Search methods based on primary sequence struggle to accurately detect ...homology between sequences with less than 20% amino acid identity. Profile- and structure-based strategies extend sensitive search capabilities into this twilight zone of sequence similarity but require slow pre-processing steps. Recently, whole-protein and positional embeddings from deep neural networks have shown promise for providing sensitive sequence comparison and annotation at long evolutionary distances. Embeddings are generally faster to compute than profiles and predicted structures but still suffer several drawbacks related to the ability of whole-protein embeddings to discriminate domain-level homology, and the database size and search speed of methods using positional embeddings. In this work, we show that low-dimensionality positional embeddings can be used directly in speed-optimized local search algorithms. As a proof of concept, we use the ESM2 3B model to convert primary sequences directly into the 3D interaction (3Di) alphabet or amino acid profiles and use these embeddings as input to the highly optimized Foldseek, HMMER3, and HH-suite search algorithms. Our results suggest that positional embeddings as small as a single byte can provide sufficient information for dramatically improved sensitivity over amino acid sequence searches without sacrificing search speed.
The predominant methodology for DNA methylation analysis relies on the chemical deamination by sodium bisulfite of unmodified cytosine to uracil to permit the differential readout of methylated ...cytosines. Bisulfite treatment damages the DNA, leading to fragmentation and loss of long-range methylation information. To overcome this limitation of bisulfite-treated DNA, we applied a new enzymatic deamination approach, termed enzymatic methyl-seq (EM-seq), to long-range sequencing technologies. Our methodology, named long-read enzymatic modification sequencing (LR-EM-seq), preserves the integrity of DNA, allowing long-range methylation profiling of 5-methylcytosine (5mC) and 5-hydroxymethylcytosine (5hmC) over multikilobase length of genomic DNA. When applied to known differentially methylated regions (DMRs), LR-EM-seq achieves phasing of >5 kb, resulting in broader and better defined DMRs compared with that previously reported. This result showed the importance of phasing methylation for biologically relevant questions and the applicability of LR-EM-seq for long-range epigenetic analysis at single-molecule and single-nucleotide resolution.
T4 RNA ligases are commonly used to attach adapters to RNAs, but large differences in ligation efficiency make detection and quantitation problematic. We developed a ligation selection strategy using ...random RNAs in combination with high-throughput sequencing to gain insight into the differences in efficiency of ligating pre-adenylated DNA adapters to RNA 3'-ends. After analyzing biases in RNA sequence, secondary structure and RNA-adapter cofold structure, we conclude that T4 RNA ligases do not show significant primary sequence preference in RNA substrates, but are biased against structural features within RNAs and adapters. Specifically, RNAs with less than three unstructured nucleotides at the 3'-end and RNAs that are predicted to cofold with an adapter in unfavorable structures are likely to be poorly ligated. The effect of RNA-adapter cofold structures on ligation is supported by experiments where the ligation efficiency of specific miRNAs was changed by designing adapters to alter cofold structure. In addition, we show that using adapters with randomized regions results in higher ligation efficiency and reduced ligation bias. We propose that using randomized adapters may improve RNA representation in experiments that include a 3'-adapter ligation step.
Ulcerative colitis (UC) and metabolic dysfunction-associated steatotic liver disease (MASLD) are closely intertwined; however, the precise molecular mechanisms governing their coexistence remain ...unclear.
We obtained UC (GSE75214) and MASLD (GSE151158) datasets from the Gene Expression Omnibus (GEO) database. Differentially expressed genes (DEGs) were acquired by the 'edgeR' and 'limma' packages of R. We then performed functional enrichment analysis of common DEGs. Hub genes were selected using the cytoHubba plugin and validated using GSE87466 for UC and GSE33814 for MASLD. Immunohistochemistry was employed to validate the hub genes' expression in clinical samples. Immune infiltration and gene set enrichment analyses of the hub genes were performed. Finally, we estimated the Spearman's correlation coefficients for the clinical correlation of the core genes.
Within a cohort of 26 differentially regulated genes in both UC and MASLD, pathways involving cytokine-mediated signaling, cell chemotaxis, and leukocyte migration were enriched. After further validation,
, and
were identified as the hub genes. Analysis of immune infiltration patterns highlighted an association between elevated pivotal gene expression and M1 macrophage activation. Immunohistochemical staining revealed widespread expression of pivotal genes in UC- and MASLD-affected tissues. Furthermore, significant correlations were observed between the increased expression of hub genes and biochemical markers, such as albumin and prothrombin time.
This bioinformatics analysis highlights
, and
as crucial genes involved in the co-occurrence of UC and MASLD, providing insights into the underlying mechanisms of these two conditions.
The smallest genomes of any photosynthetic organisms are found in a group of free-living marine cyanobacteria, Prochlorococcus. To determine the underlying evolutionary mechanisms, we developed a new ...method to reconstruct the steps leading to the Prochlorococcus genome reduction using 12 Prochlorococcus and 6 marine Synechococcus genomes. Our results reveal that small genome sizes within Prochlorococcus were largely determined shortly after the split of Prochlorococcus and Synechococcus (an early big shrink) and thus for the first time decouple the genome reduction from Prochlorococcus diversification. A maximum likelihood approach was then used to estimate changes of nucleotide substitution rate and selection strength along Prochlorococcus evolution in a phylogenetic framework. Strong genome wide purifying selection was associated with the loss of many genes in the early evolutionary stage. The deleted genes were distributed around the genome, participated in many different functional categories and in general had been under relaxed selection pressure. We propose that shortly after Prochlorococcus diverged from its common ancestor with marine Synechococcus, its population size increased quickly thus increasing efficacy of selection. Due to limited nutrients and a relatively constant environment, selection favored a streamlined genome for maximum economy. Strong genome wide selection subsequently caused the loss of genes with small functional effect including the loss of some DNA repair genes. In summary, genome reduction in Prochlorococcus resulted in genome features that are similar to symbiotic bacteria and pathogens, however, the small genome sizes resulted from an increase in genome wide selection rather than a consequence of a reduced ecological niche or relaxed selection due to genetic drift.
Due to the fact that intelligent algorithms such as Particle Swarm Optimization (PSO) and Differential Evolution (DE) are susceptible to local optima and the efficiency of solving an optimal solution ...is low when solving the optimal trajectory, this paper uses the Sequential Quadratic Programming (SQP) algorithm for the optimal trajectory planning of a hydraulic robotic excavator. To achieve high efficiency and stationarity during the operation of the hydraulic robotic excavator, the trade-off between the time and jerk is considered. Cubic splines were used to interpolate in joint space, and the optimal time-jerk trajectory was obtained using the SQP with joint angular velocity, angular acceleration, and jerk as constraints. The optimal angle curves of each joint were obtained, and the optimal time-jerk trajectory planning of the excavator was realized. Experimental results show that the SQP method under the same weight is more efficient in solving the optimal solution and the optimal excavating trajectory is smoother, and each joint can reach the target point with smaller angular velocity, and acceleration change, which avoids the impact of each joint during operation and conserves working time. Finally, the excavator autonomous operation becomes more stable and efficient.
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