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This study aimed to explore the selection of induction chemotherapy (IC) cycles for stage N3 nasopharyngeal carcinoma (NPC). We employed propensity score matching (PSM) to categorize patients into 3-cycle and 4-cycle IC groups (IC = 3 and IC = 4). The log-rank and chi-squared tests were used respectively to evaluate the differences in survival and acute toxicities. Survival outcomes including overall survival (OS), progression-free survival (PFS), distant metastasis-free survival (DMFS), and locoregional relapse-free survival (LRRFS) were evaluated among the two groups. After PSM, each group comprised 99 patients. The IC = 4 group exhibited markedly improved survival outcomes compared with the IC = 3 group. Multivariate analysis revealed that pre-EBV DNA was an independent risk factor affecting PFS and DMFS. For high-risk patients with pre-EBV DNA ≥ 7800 copies/ml, the IC = 4 group demonstrated greater survival compared to the IC = 3 group. Among low-risk patients with pre-EBV DNA < 7800 copies/ml, both groups showed comparable survival outcomes. In terms of acute adverse reactions, the IC = 4 group experienced higher incidences, particularly with grade 2–4 alanine transaminase elevation and thrombocytopenia. For stage N3 NPC, pre-EBV DNA could be a powerful predictor for guiding the selection of IC cycles. The IC = 4 regimen is probably more beneficial to high-risk patients due to superior survival, while for low-risk patients, the IC = 3 regimen may be sufficient. [ABSTRACT FROM AUTHOR]

The rapid rise of live-streaming selling and the increasing diversity of e-commerce channels have sparked ongoing debates regarding product pricing power. This price competition behaviour of online dual channels triggered by the supply-side pricing dilemma is a topic of significant research interest and forms the focal point of this paper. Therefore, we leverage the principles of game theory to construct a three-party evolutionary game model consisting of a brand, a steamer, and a traditional e-commerce platform. Within this framework, the brand holds the primary pricing authority. Our analysis examines the equilibrium strategies adopted by the various players under diverse scenarios and explores how the evolving behaviours of these parties impact the overall game system. Additionally, we employ the "people-goods-scene" perspective to deconstruct the intricate dynamics within this complex supply chain ecosystem. Our findings reveal two distinct evolutionary stable strategies: one in which brands maintain a stance against low-price negotiation, streamers avoid bargaining, and platforms avoid providing subsidies; the other in which brands are open to negotiating low prices, streamers engage in bargaining, and platforms provide subsidies. Notably, in most cases, brands tend to avoid explicitly stating their opposition to low-price negotiation. However, if a brand's strategy shifts towards rejecting such negotiation, the negative implications of price wars across channels become a critical consideration. Conversely, when a brand opts for a more concessive pricing strategy, we need to consider the potential positive impacts of low-price competition. [ABSTRACT FROM AUTHOR]

In order to study the progressive collapse resistance of plate-member composite reticulated shell structure and propose an applicable analysis method, the complete and member removal experimental models of composite reticulated shell structure were designed by Alternate Path (AP) analysis method, and the progressive collapse resistance test and numerical analysis were carried out. The results show that the honeycomb plate contributes greatly to the bearing capacity of the composite structure, the ultimate bearing capacity of member removal structure is 3.64% smaller than that of the complete structure, and the ultimate displacement is 30% smaller. The average error of load-displacement curves between the progressive collapse resistance test and numerical analysis is less than 10% in the elastic stage, and the overall failure mode of the numerical analysis model and the experimental model is consistent. The anti-progressive collapse analysis method based on modal sensitivity is suitable for the plate-member composite reticulated shell structure due to high computational efficiency and accuracy, which provides a reference for progressive collapse resistance analysis of plate-member composite reticulated shell structure. [ABSTRACT FROM AUTHOR]

The objectives of the study were to systematically evaluate the rehabilitation effect of noninvasive brain stimulation (NIBS) on upper extremity motor function and activities of daily living in stroke patients and to prioritize various stimulation protocols for reliable evidence-based medical recommendations in patients with upper extremity motor dysfunction after stroke. Web of Science, PubMed, Embase, Cochrane Library, CNKI, Wanfang, VIP, and CBM were searched to collect all randomized controlled trials (RCTs) of NIBS to improve upper extremity motor function in stroke patients. The retrieval time was from the establishment of all databases to May 2023. According to the Cochrane system evaluation manual, the quality of the included studies was evaluated, and the data were extracted. Statistical analysis was carried out by using RevMan 5.3, R 4.3.0, and Stata 17.0 software. Finally, 94 RCTs were included, with a total of 5546 patients. Meta-analysis showed that NIBS improved the Fugl-Meyer assessment (FMA) score (mean difference (MD) = 6.51, 95% CI 6.20 ~ 6.82, P < 0.05), MBI score (MD = 7.69, 95% CI 6.57 ~ 8.81, P < 0.05), ARAT score (MD = 5.06, 95% CI 3.85 ~ 6.27, P < 0.05), and motor evoked potential (MEP) amplitude. The modified Ashworth scale score (MD = − 0.37, 95% CI − 0.60 to − 0.14, P < 0.05), National Institutes of Health Stroke Scale score (MD = − 2.17, 95% CI − 3.32 to − 1.11, P < 0.05), incubation period of MEP (MD = − 0.72, 95% CI − 1.06 to − 0.38, P < 0.05), and central motor conduction time (MD = − 0.90, 95% CI − 1.29 to − 0.50, P < 0.05) were decreased in stroke patients. Network meta-analysis showed that the order of interventions in improving FMA scores from high to low was anodal-transcranial direct current stimulation (tDCS) (surface under the cumulative ranking curve (SUCRA) = 83.7%) > cathodal-tDCS (SUCRA = 80.2%) > high-frequency (HF)-repetitive transcranial magnetic stimulation (rTMS) (SUCRA = 68.5%) > low-frequency (LF)-rTMS (SUCRA = 66.5%) > continuous theta burst stimulation (cTBS) (SUCRA = 54.2%) > bilateral-tDCS (SUCRA = 45.2%) > intermittent theta burst stimulation (iTBS) (SUCRA = 34.1%) > sham-NIBS (SUCRA = 16.0%) > CR (SUCRA = 1.6%). In terms of improving MBI scores, the order from high to low was anodal-tDCS (SUCRA = 88.7%) > cathodal-tDCS (SUCRA = 85.4%) > HF-rTMS (SUCRA = 63.4%) > bilateral-tDCS (SUCRA = 56.0%) > LF-rTMS (SUCRA = 54.2%) > iTBS (SUCRA = 32.4%) > sham-NIBS (SUCRA = 13.8%) > CR (SUCRA = 6.1%). NIBS can effectively improve upper extremity motor function and activities of daily living after stroke. Among the various NIBS protocols, anodal-tDCS demonstrated the most significant intervention effect, followed by cathodal-tDCS and HF-rTMS. [ABSTRACT FROM AUTHOR]

Here, we report a novel ourmia-like mycovirus, named “Phomopsis asparagi magoulivirus 1” (PaMV1), derived from the phytopathogenic fungus Phomopsis asparagi. The genome of PaMV1 consists of a positive-sense single-stranded RNA (+ ssRNA) that is 2,639 nucleotides in length, with a GC content of 57.13%. It contains a single open reading frame (ORF) encoding a putative RNA-dependent RNA polymerase (RdRp) consisting of 686 amino acids with a molecular mass of 78.57 kDa. Phylogenetic analysis based on RdRp sequences revealed that PaMV1 grouped together with Diaporthe gulyae magoulivirus 1 (DgMV1) in a distinct clade. Sequence comparisons and phylogenetic analysis suggest that PaMV1 is a novel member of the genus Magoulivirus, family Botourmiaviridae. [ABSTRACT FROM AUTHOR]

Cerebral small vessel disease (CSVD) is the most common progressive vascular disease that causes vascular dementia. Aging and hypertension are major contributors to CSVD, but the pathophysiological mechanism remains unclear, mainly due to the lack of an ideal animal model. Our previous study revealed that vascular smooth muscle cell (VSMC)-specific myosin phosphatase target subunit 1 (MYPT1) knockout (MYPT1SMKO) leads to constant hypertension, prompting us to explore whether hypertensive MYPT1SMKO mice can be considered a novel CSVD animal model. Here, we found that MYPT1SMKO mice displayed age-dependent CSVD-like neurobehaviors, including decreased motion speed, anxiety, and cognitive decline. MYPT1SMKO mice exhibited remarkable white matter injury compared with control mice, as shown by the more prominent loss of myelin at 12 months of age. Additionally, MYPT1SMKO mice were found to exhibit CSVD-like small vessel impairment, including intravascular hyalinization, perivascular space enlargement, and microbleed and blood-brain barrier (BBB) disruption. Last, our results revealed that the brain of MYPT1SMKO mice was characterized by an exacerbated inflammatory microenvironment, which is similar to patients with CSVD. In light of the above structural and functional phenotypes that closely mimic the conditions of human CSVD, we suggest that MYPT1SMKO mice are a novel age- and hypertension-dependent animal model of CSVD. [ABSTRACT FROM AUTHOR]

The study delves into the behavior of cos-Gaussian breathers navigating through a highly nonlocal medium, with a spotlight on their width and intensity variations under different parameter adjustments. The mathematical formulas describing the characteristics of cos-Gaussian beams have been derived. By controlling the parameters, the transverse width of cos-Gaussian beams can exhibit various variations, reflecting various mode transformations. We scrutinized how alterations in cosine parameters, offset distance, and initial energy manifest in the breather's width and shape during propagation. Our observations underscore the significant influence of these parameters on the dynamic evolution and morphology of cos-Gaussian breathers, thereby unveiling the potential application of using parameters to control the dynamic characteristics of cos-Gaussian beam transmission. [ABSTRACT FROM AUTHOR]

Chronic cerebral hypoperfusion is one of the pathophysiological mechanisms contributing to cognitive decline by causing white matter injury. Microglia phagocytosing myelin debris in a timely manner can promote remyelination and contribute to the repair of white matter. However, milk fat globule-epidermal growth factor-factor 8 (MFG-E8), a microglial phagocytosis-related protein, has not been well studied in hypoperfusion-related cognitive dysfunction. We found that the expression of MFG-E8 was significantly decreased in the brain of mice after bilateral carotid artery stenosis (BCAS). MFG-E8 knockout mice demonstrated more severe BCAS-induced cognitive impairments in the behavioral tests. In addition, we discovered that the deletion of MFG-E8 aggravated white matter damage and the destruction of myelin microstructure through fluorescent staining and electron microscopy. Meanwhile, MFG-E8 overexpression by AAV improved white matter injury and increased the number of mature oligodendrocytes after BCAS. Moreover, in vitro and in vivo experiments showed that MFG-E8 could enhance the phagocytic function of microglia via the αVβ3/αVβ5/Rac1 pathway and IGF-1 production to promote the differentiation of oligodendrocyte progenitor cells into mature oligodendrocytes. Interestingly, we found that MFG-E8 was mainly derived from astrocytes, not microglia. Our findings suggest that MFG-E8 is a potential therapeutic target for cognitive impairments following cerebral hypoperfusion. [ABSTRACT FROM AUTHOR]

Moyamoya disease (MMD) is a chronic occlusive cerebrovascular disease with the development of a network of abnormal vessels. Immune inflammation is associated with the occurrence and development of MMD. However, the mechanisms underlying the formation of the abnormal vascular network remain unclear. Twenty-eight patients with MMD, 26 ischemic stroke patients, and 26 unrelated healthy volunteers were enrolled in this study The data showed that the levels of granulocyte-macrophage colony-stimulating factor (GM-CSF) were higher in MMD patients than in healthy controls (P <0.01), and GM-CSF was mainly from Th1 and Th17 cells in MMD. We found that increased GM-CSF drove monocytes to secrete a series of cytokines associated with angiogenesis, inflammation, and chemotaxis. In summary, our findings demonstrate for the first time the important involvement of GM-CSF in MMD and that GM-CSF is an important factor in the formation of abnormal vascular networks in MMD. [ABSTRACT FROM AUTHOR]

To explore the livelihood status and key influencing factors of rural households in the minority areas, we collected flat data from 284 rural households in 32 villages across 12 counties of Western Sichuan from 2021 to 2022. We conducted participatory household survey on the livelihood status of the rural households and try to identify the key factors to influence their livelihood vulnerability using multiple linear regression. The results showed that: the livelihood situation of the rural households is relatively vulnerable. The vulnerability varies significantly with the income levels, education levels, and income sources. The vulnerability of farm households, categorized from low to high livelihood types, follows the sequence: non-agricultural dominant households, non-agricultural households, agricultural dominant households, and pure agricultural households. The degree of damage to the natural environment, education costs, loan opportunities, the proportion of agricultural income to annual household income, and the presence of sick people in the household have significant positive effects on the livelihood vulnerability index (LVI) of rural households; while help from relatives and friends, net income per capita, household size, household education, agricultural land area, participation in industrial organizations, number of livestock, purchase of commercial houses, drinking water source, and self-supply of food have significant negative effects. Based on the findings, we believe that local rural households operate in a complex livelihood system and recommend continuous interventions targeting key influences to provide empirical research support for areas facing similar situations. [ABSTRACT FROM AUTHOR]

Long-term electroencephalogram (Long-Term EEG) has the capacity to monitor over a long period, making it a valuable tool in medical institutions. However, due to the large volume of patient data, selecting clean data segments from raw Long-Term EEG for further analysis is an extremely time-consuming and labor-intensive task. Furthermore, the various actions of patients during recording make it difficult to use algorithms to denoise part of the EEG data, and thus lead to the rejection of these data. Therefore, tools for the quick rejection of heavily corrupted epochs in Long-Term EEG records are highly beneficial. In this paper, a new reliable and fast automatic artifact rejection method for Long-Term EEG based on Isolation Forest (IF) is proposed. Specifically, the IF algorithm is repetitively applied to detect outliers in the EEG data, and the boundary of inliers is promptly adjusted by using a statistical indicator to make the algorithm proceed in an iterative manner. The iteration is terminated when the distance metric between clean epochs and artifact-corrupted epochs remains unchanged. Six statistical indicators (i.e., min, max, median, mean, kurtosis, and skewness) are evaluated by setting them as centroid to adjust the boundary during iteration, and the proposed method is compared with several state-of-the-art methods on a retrospectively collected dataset. The experimental results indicate that utilizing the min value of data as the centroid yields the most optimal performance, and the proposed method is highly efficacious and reliable in the automatic artifact rejection of Long-Term EEG, as it significantly improves the overall data quality. Furthermore, the proposed method surpasses compared methods on most data segments with poor data quality, demonstrating its superior capacity to enhance the data quality of the heavily corrupted data. Besides, owing to the linear time complexity of IF, the proposed method is much faster than other methods, thus providing an advantage when dealing with extensive datasets. [ABSTRACT FROM AUTHOR]

Idiopathic pulmonary fibrosis (IPF) is a chronic pulmonary fibrosis disease that is fatal. Mesenchymal stem cells (MSCs)-secreted exosomes (exos) have been linked to improving PF. Moreover, exosomal microRNAs (miRs) can control the growth of numerous diseases, including lung disorders. Our bioinformatics analysis showed that miR-30b was downregulated in tissue samples from surgical remnants of biopsies or lungs explanted from patients with IPF who underwent pulmonary transplantation. This suggests that miR-30b plays an important role in both the pathogenesis and treatment of IPF. Herein, this research was designed to ascertain the mechanism of MSCs-exos-packaged miR-30b in alleviating PF. The serum was harvested from idiopathic PF (IPF) patients with interstitial pneumonia caused by dermatomyositis and the MLE12 lung epithelial cell fibrosis model was built with TGF-β1 (10 ng/mL), followed by miR-30b expression determination. TGF-β1-stimulated MLE12 cells were co-incubated with exos from MSCs with or without Spred2 or Runx1 overexpression, followed by measurement of cell viability and apoptosis. After establishing the IPF mouse model with bleomycin and injecting exos and/or silencing and overexpressing adenovirus vectors, fibrosis evaluation was conducted. In mice and cells, the expression of TGF-β1, TNF-α, and IL-1β was tested via ELISA, and the levels of E-cad, ZO-1, α-SMA, and collagen type I via western blot analysis. The promoters of miR-30b, Runx1, and Spred2 were investigated. miR-30b was downregulated in the serum of IPF patients and TGF-β1-stimulated MLE12 cells. Mechanistically, miR-30b inhibited Spred2 transcription by negatively targeting Runx1. MSCs-exos or MSCs-exo-miR-30b decreased the apoptosis, inflammation, and fibrosis while increasing their viability in TGF-β1-stimulated MLE12 cells, which was annulled by overexpressing Runx1 or Spred2. Exo-miR-30b decreased Runx1 expression to downregulate Spred2, reducing fibrosis and inflammation in IPF mice. Our results indicated that MSCs-exos-encapsulated miR-30b had a potential function to inhibit PF and part of its function may be achieved by targeting RUNX1 to reduce the Spred2 transcription level. Moreover, this work offered evidence and therapeutic targets for therapeutic strategies for managing clinical PF in patients. [ABSTRACT FROM AUTHOR]

The TMEM63 family proteins (A, B, and C), calcium-permeable channels in animals that are preferentially activated by hypo-osmolality, have been implicated in various physiological functions. Deficiency of these channels would cause many diseases including hearing loss. However, their structures and physiological roles are not yet well understood. In this study, we determine the cryo-electron microscopy (cryo-EM) structure of the mouse TMEM63C at 3.56 Å, and revealed structural differences compared to TMEM63A, TMEM63B, and the plant orthologues OSCAs. Further structural guided mutagenesis and calcium imaging demonstrated the important roles of the coupling of TM0 and TM6 in channel activity. Additionally, we confirm that TMEM63C exists primarily as a monomer under physiological conditions, in contrast, TMEM63B is a mix of monomer and dimer in cells, suggesting that oligomerization is a regulatory mechanism for TMEM63 proteins. TMEM63s are mechanosensitive ion channels activated by hypo-osmolality. Here, the authors determine the cryo-EM structure of mouse TMEM63C confirming its predominant monomeric state and the significance of TM0-TM6 coupling in channel activity. [ABSTRACT FROM AUTHOR]

Intracranial hemorrhage (ICH) is a neurological emergency with a poor prognosis. This study aimed to understand the association between hemoglobin levels, red blood cell distribution width ratio (HRR), and mortality in patients with ICH. Information on patients with ICH was extracted from the Medical Information Mart for Intensive Care IV (MIMIC-IV) database. Cox proportional risk models were used to assess the relationship between HRR and 28-day and 90-day mortality, and constructed by adjusting for relevant covariates. Segmented regression models and smoothing curves were used to analyze the linear relationship between HRR and mortality. This study recruited 4,716 patients with ICH. The HRR Q4 group was negatively associated with the 28- and 90-day mortality. For patients aged > 65 years, the right-hand threshold inflection points of the HRR were 0.92 and 0.93, respectively, which were negatively associated with 28-day mortality (HR:0.06, 95% CI 0.01, 0.35, p = 0.0016) and with 90-day mortality (p = 0.0006). In the non-linear model, both 28-day mortality (HR, 0.17; 95% CI 0.04–0.75, p = 0.0191; HRR > 0.89) and 90-day mortality (HR, 0.13; 95% CI 0.04–0.49, p = 0.0022; HRR > 0.85) were associated in men. In the subgroup analysis, the negative association between HRR and mortality was more pronounced in patients > 65 years of age, as well as in patients with non-dementia, diabetes, and malignant cancer. We found a non-linear relationship between mortality and the HRR in elderly patients, and a higher HRR was negatively associated with mortality in patients with ICH. [ABSTRACT FROM AUTHOR]

Purpose: We aimed to analyse the correlation between prostaglandin D2 synthase (PTGDS) and a diffuse large B-cell lymphoma (DLBCL) prognosis. Methods: We retrospectively collected two hundred paraffin-embedded tissue specimens that were pathologically diagnosed as DLBCL in the Fujian Tumour Hospital between January 2014 and December 2018. An abundance of paraffin-embedded tumour tissues were obtained. Twenty patients with lymphocyte-rich, benign, tissue-reactive, hypertrophic tonsillitis were selected as controls. Wax blocks were selected for primary cases and the controls were screened by professional pathologists. The levels of prostaglandin D2 synthase (PTGDS) and the EMT-related molecules, E-cadherin and vimentin, were detected by immunohistochemistry in clinical samples. A chi-square test revealed the correlations between PTGDS expression and clinicopathological characteristics, including age, sex, primary site, clinical stage, immunotyping, and International Prognostic Index (IPI) score. The Kaplan–Meier survival analysis was performed, and the diagnostic value was evaluated by receiver operating characteristic (ROC) curve analysis. Results: A total of 138 cases (69%) were found to be PTGDS positive (> 30% positive cells). PTGDS staining was negative (< 30% positive cells) in 62 cases (31%). We collected the corresponding clinicopathological information and found that PTGDS expression was not significantly related to the patients' age, tumour stage, presence of extranodal invasion, or IPI score. According to the follow-up data, patients with low PTGDS expression had poor progression-free survival (PFS) and overall survival (OS), with 2-year PFS and OS rates of 41.7% and 50%, respectively. The 2-year PFS and OS rates of PTGDS-positive patients were 89.3% and 92.9%, respectively (P < 0.0001), and the differences were significant. Conclusion: We found that the expression level of PTGDS is significantly correlated with the prognosis of diffuse large B-cell lymphoma. [ABSTRACT FROM AUTHOR]

Lipid droplets (LDs) were reported to play an important role in the modulation of inflammation and various cellular processes among multiple cell types. However, LDs accumulation, its function and mechanisms of its formation during ischemic stroke remained poorly-identified. In this study, we observed increased LDs accumulation in microglia at the acute stage of ischemic stroke by immunofluorescence and flow cytometry. Transcriptomic analysis indicated that microglia accumulated with LDs were associated with inflammation and phagocytosis. Both inflammatory activation and phagocytosis of tissue debris in microglia could contribute to LDs formation. Moreover, through specific LDs depletion and overload experiments by pharmacological approaches, we proposed that LDs was critical for the maintenance of anti-inflammatory properties of microglia. Furthermore, Atglistatin, a specific adipose triglyceride lipase (ATGL) inhibitor, was shown to prevent proinflammatory cytokines production in primary microglia through decreased LDs lipolysis. After Atglistatin treatment, middle cerebral artery occlusion (MCAO) mice showed decreased infarct volume and improved neurobehavioral performance at the acute stage of stroke. Our findings provided a biological basis for microglial LDs regulation as a potential therapeutic strategy for acute ischemic stroke and uncovered the neuroprotective role of Atglistatin in the treatment of MCAO mice. [ABSTRACT FROM AUTHOR]

Chronic cerebral hypoperfusion leads to white matter injury (WMI), which subsequently causes neurodegeneration and even cognitive impairment. However, due to the lack of treatment specifically for WMI, novel recognized and effective therapeutic strategies are urgently needed. In this study, we found that honokiol and magnolol, two compounds derived from Magnolia officinalis, significantly facilitated the differentiation of primary oligodendrocyte precursor cells (OPCs) into mature oligodendrocytes, with a more prominent effect of the former compound. Moreover, our results demonstrated that honokiol treatment improved myelin injury, induced mature oligodendrocyte protein expression, attenuated cognitive decline, promoted oligodendrocyte regeneration, and inhibited astrocytic activation in the bilateral carotid artery stenosis model. Mechanistically, honokiol increased the phosphorylation of serine/threonine kinase (Akt) and mammalian target of rapamycin (mTOR) by activating cannabinoid receptor 1 during OPC differentiation. Collectively, our study indicates that honokiol might serve as a potential treatment for WMI in chronic cerebral ischemia. [ABSTRACT FROM AUTHOR]

Circular RNAs have been extensively studied in eukaryotes, but their presence and/or biological functionality in bacteria are unclear. Here, we show that a regulatory noncoding RNA (DucS) exists in both linear and circular conformation in Bacillus altitudinis. The linear forms promote B. altitudinis tolerance to H2O2 stress, partly through increased translation of a stress-responsive gene, htrA. The 3′ end sequences of the linear forms are crucial for RNA circularization, and formation of circular forms can decrease the levels of the regulatory linear cognates. Bioinformatic analysis of available RNA-seq datasets from 30 bacterial species revealed multiple circular RNA candidates, distinct from DucS, for all the examined species. Experiments testing for the presence of selected circular RNA candidates in four species successfully validated 7 out of 9 candidates from B. altitudinis and 4 out of 5 candidates from Bacillus paralicheniformis; However, none of the candidates tested for Bacillus subtilis and Escherichia coli were detected. Our work identifies a dual-conformation regulatory RNA in B. altitutidinis, and indicates that circular RNAs exist in diverse bacteria. However, circularization of specific RNAs does not seem to be conserved across species, and the circularization mechanisms and biological functionality of the circular forms remain unclear. The presence and/or biological functionality of circular RNAs in bacteria are unclear. Here, the authors identify a dual-conformation (linear and circular) noncoding RNA that promotes tolerance to oxidative stress in Bacillus altitutidinis, and provide evidence for the existence of other circular RNAs in diverse bacterial species. [ABSTRACT FROM AUTHOR]

We report a copper-catalyzed ligand-controlled selective 1,2- and 1,4-hydrosilylation of 1,3-enynes, which furnishes enantiomerically enriched propargyl- and 1,2-allenylsilane products in high yields with excellent enantioselectivities (up to 99% ee). This reaction proceeds under mild conditions, shows broad substrate scope for both 1,3-enynes and trihydrosilanes, and displays excellent regioselectivities. Mechanistic studies based on deuterium-labeling reactions and density functional theory (DFT) calculations suggest that allenylcopper is the dominant reactive intermediate under both 1,2- and 1,4-hydrosilylation conditions, and it undergoes metathesis with silanes via selective four-membered or six-membered transition state, depending on the nature of the ligand. The weak interactions between the ligands and the reacting partners are found to be the key controlling factor for the observed regioselectivity switch. The origin of high enantiocontrol in the 1,4-hydrosilylation is also revealed by high level DLPNO-CCSD(T) calculations. Functionalization of 1,3-enyne represent a powerful method to construct allenyl- and propargylderivatives, but hydrosilylation reaction of 1,3-enynes remains underdeveloped. Here, the authors report a copper-catalyzed ligand-controlled selective 1,2- and 1,4- hydrosilylation of 1,3-enynes which proceeds enantioselectively with high yields. [ABSTRACT FROM AUTHOR]

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For decades, the preparation of polyisoprene rubber that can match the comprehensive properties of natural rubber (NR) has been pursued. While sacrificial bonds have been used to promote the strength and toughness of rubbers, little is known about their effects on fatigue resistance, which is important in dynamic environments. Herein, terminal block and randomly functionalized polyisoprene rubbers tethered with di-alanine, tri-alanine and tetra-alanine were prepared. The results showed that the flow activation energy, aggregates ordering and energy dissipation of the hydrogen-bonded aggregates increase with the elongation of oligopeptide length (XA, X=2, 3, 4), therefore resulting in enhanced mechanical strength and toughness of corresponding samples. Comparably, the tear strengths are barely affected by oligopeptide lengths in block samples, but promoted from dipeptide to tetrapeptide in random samples, probably due to the well dispersed oligopeptide aggregates. Most importantly, it is found that the tight binding aggregates of oligopeptides are critical for the excellent fatigue resistance, which is absent in polyisoprene and natural rubber. The loose aggregates dissociate and recombine repeatedly under cyclic loading and the tight aggregates keep the network integrated and robust. Interestingly, the largest hysteresis of PIP-4A-V with the longest oligopeptide length give the lowest heat generation, which is contrary to the traditional sacrificial bonds. Overall, the oligopeptide aggregates have repeatable energy dissipation properties and cycle life comparable to or even surpassing those of the linked proteins in NR, resulting in similar tensile strength, fracture toughness, and better fatigue resistance relative to NR. This deep insight on the role of oligopeptide aggregates is very useful for the engineering rubbers served in dynamic environments. [ABSTRACT FROM AUTHOR]

A numerical simulation method based on CFD has been established to simulate the fully coupled motion for an attenuator-type wave energy converter (WEC). Based on this method, a detailed parametric analysis has been conducted to investigate the design of the rafts. The effects of different parameters (wave parameters, structural parameters and PTO parameters) on the hydrodynamic characteristics of the attenuator-type WEC were studied in detail. The results show that in terms of wave parameters, there is an optimal wave period, which makes the relative pitching angle amplitude of the WEC reach the maximum, and the increase of wave height is conducive to the relative pitching angle amplitude of wave energy. Under different wave conditions, the relative pitch angle of the parallelogram raft device is the maximum. In terms of structural parameters, the parallelogram attenuator-type device has the optimal values in different relative directions, different distances and different apex angle, which makes the relative motion amplitude of the device reach the maximum, and the spacing and the apex angle have influence on the motion frequency of the device, while the relative direction has almost no influence on it. In terms of PTO parameters, there is an optimal damping coefficient, which makes the power generation efficiency of the WEC reach the maximum. The research results provide a valuable reference for future research and design of the attenuator-type WEC. [ABSTRACT FROM AUTHOR]

The crystal structure, phase transition, metamagnetic behavior and magnetic entropy change (ΔSM) of the Mn49−xCuxNi41Sn10(x = 0–2) alloys were systematically studied. The results of experiment about crystal structure confirm that Mn49−xCuxNi41Sn10 alloys have a highly ordered cubic L21 Heusler structure. With Cu element improving, all samples have a martensitic transformation and inverse martensitic transformation by analyzing the result of magnetic measurement. The saturation magnetization of samples decreases gradually, the martensitic transformation temperature (TM) of as-annealed samples increases to a high-temperature area, and the Curie temperature (TC) of samples moves to a low-temperature area with the content of Cu element increasing from x = 0 to x = 2. The positive values of magnetic entropy change (ΔSM) of all alloys were investigated near the martensitic transformation. When the applied magnetic field is 3 T, a worthwhile value of ΔSM can reach up to 32.0 J·kg−1·K−1 in Mn47Cu2Ni41Sn10 alloy. [ABSTRACT FROM AUTHOR]

The production and composition of extracellular polymeric substances (EPS), as well as the EPS-related functional resistance genes and metabolic levels of Bacillus sp. under Cu(II) stress, were investigated. EPS production increased by 2.73 ± 0.29 times compared to the control when the strain was treated with 30 mg L−1 Cu(II). Specifically, the polysaccharide (PS) content in EPS increased by 2.26 ± 0.28 g CDW−1 and the PN/PS (protein/polysaccharide) ratio value increased by 3.18 ± 0.33 times under 30 mg L−1 Cu(II) compared to the control. The increased EPS secretion and higher PN/PS ratio in EPS strengthened the cells' ability to resist the toxic effect of Cu(II). Differential expression of functional genes under Cu(II) stress was revealed by Gene Ontology pathway enrichment analysis and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis. The enriched genes were most obviously upregulated in the UMP biosynthesis pathway, the pyrimidine metabolism pathway, and the TCS metabolism pathway. This indicates an enhancement of EPS regulation-related metabolic levels and their role as a defense mechanism for cells to adapt to Cu(II) stress. Additionally, seven copper resistance genes were upregulated while three were downregulated. The upregulated genes were related to the heavy metal resistance, while downregulated genes were related to cell differentiation, indicating that the strain had initiated an obvious resistance to Cu(II) despite its severe cell toxicity. These results provided a basis for promoting EPS-regulated associated functional genes and the application of gene-regulated bacteria in heavy metal-containing wastewater treatment. [ABSTRACT FROM AUTHOR]

In geophysical exploration, the expression of the electromagnetic field in homogeneous layered media involves Hankel integrals, and the kernel function can be simplified into a combination of zero- and first-order Bessel functions. Owing to the complex nature of the electromagnetic field, only a few analytical expressions for the Hankel integrals of the electromagnetic field in layered media exist. Moreover, as the argument of the Bessel functions increases, the Hankel integrals exhibit rapid oscillations and slow decay, making it difficult for the computation of the electromagnetic field to achieve high accuracy, especially for high-frequency and long-range transmission and reception scenarios. To address this issue, we propose a fully analytical integration method applicable to both high and low frequencies and long and short transmission and reception distances. The study used quadratic function interpolation for the coefficient part of the Hankel integrals in the expression of the electromagnetic field in layered media. The interpolated results within each element, multiplied by the Bessel functions, could derive analytical expressions. Then, by summing up all the discrete elements, the numerical solution of the Hankel integrals could be obtained, resulting in a high-precision numerical simulation of the layered media. The fully analytical integration algorithm was tested using the expressions of the electromagnetic field in the entire space with different forms of Bessel function integrals. The computational results demonstrated the correctness and high accuracy of the algorithm. The proposed algorithm exhibited high computational accuracy in different frequency ranges and transmission and reception distances, especially in high-frequency and long-distance scenarios, where its accuracy exceeds that of the four selected digital filtering algorithms by over two orders of magnitude, indicating its strong universality.. [ABSTRACT FROM AUTHOR]

Stroke usually causes prolonged or lifelong disability, owing to the permanent loss of infarcted tissue. Although a variety of stem cell transplantation has been explored to improve neuronal defect behavior by enhancing neuroplasticity, it remains unknown whether the infarcted tissue can be reconstructed. We here cultured human cerebral organoids derived from human pluripotent stem cells (hPSCs) and transplanted them into the junction of the infarct core and the peri-infarct zone of NOD-SCID mice subjected to stroke. Months later, we found that the grafted organoids survived well in the infarcted core, differentiated into target neurons, repaired infarcted tissue, sent axons to distant brain targets, and integrated into the host neural circuit and thereby eliminated sensorimotor defect behaviors of stroke mice, whereas transplantation of dissociated single cells from organoids failed to repair the infarcted tissue. Our study offers a new strategy for reconstructing infarcted tissue via organoids transplantation thereby reversing stroke-induced disability. [ABSTRACT FROM AUTHOR]

The study aimed to investigate the consistency and diversity between metabolic and structural brain networks at individual level constructed with divergence-based method in healthy Chinese population. The 18F-FDG PET and T1-weighted images of brain were collected from 209 healthy participants. The Jensen-Shannon divergence (JSD) was used to calculate metabolic or structural connectivities between any pair of brain regions and then individual brain networks were constructed. The global and regional topological properties of both networks were analyzed with graph theoretical analysis. Regional properties including nodal efficiency, degree, and betweenness centrality were used to define hub regions of networks. Cross-modality similarity of brain connectivity was analyzed with differential power (DP) analysis. The default mode network (DMN) had the largest number of brain connectivities with high DP values. The small-worldness indexes of metabolic and structural networks in all participants were greater than 1. The structural network showed higher assortativity and local efficiency than metabolic network, while hierarchy and global efficiency were greater in the metabolic network (all P < 0.001). Most of hubs in both networks were symmetrically spatial distributed in the regions of the DMN and subcortical nuclei including thalamus and amygdala, etc. The human brain presented small-world architecture both in perspective of individual metabolic and structural networks. There was a structural substrate that supported the brain to globally and efficiently integrate and process metabolic interaction across brain regions. The cross-modality cooperation or specialization in both networks might imply mechanisms of achieving higher-order brain functions. [ABSTRACT FROM AUTHOR]

We propose a family of mixed finite elements that are robust for the nearly incompressible strain gradient model, which is a fourth-order singular perturbed elliptic system. The element is similar to [C. Taylor and P. Hood, Comput. & Fluids, 1(1973), 73–100] in the Stokes flow. Using a uniform discrete B-B inequality for the mixed finite element pairs, we show the optimal rate of convergence that is robust in the incompressible limit. By a new regularity result that is uniform in both the materials parameter and the incompressibility, we prove the method converges with 1/2 order to the solution with strong boundary layer effects. Moreover, we estimate the convergence rate of the numerical solution to the unperturbed second-order elliptic system. Numerical results for both smooth solutions and the solutions with sharp layers confirm the theoretical prediction. [ABSTRACT FROM AUTHOR]

In this work, we develop a new type of mesoporous 2D N, B, and P codoped carbon network (NBP-CNW) arranged into high-order 3D nanotube arrays (NTAs), which are wrapped onto a flexible carbon fiber microelectrode, and this microelectrode is employed as a high-performance carbon-based nanocatalyst for electrochemical biosensing. The NBP-CNW-NTAs synthesized by a facile, controllable, ecofriendly and sustainable template strategy using ionic liquids as precursors possess a high structural stability, large surface area, abundant active sites, and effective charge transport pathways, which dramatically improve their electrocatalytic activity and durability in the redox reaction of cancer biomarker H2O2. Benefiting from these unique structural merits, superb electrochemical activity and good biocompatibility, the NBP-CNW-NTAs-modified microelectrode demonstrates excellent sensing performance toward H2O2 and is embedded in a homemade microfluidic electrochemical biosensor chip for the real-time tracking of H2O2 secreted from different live cancer cells with or without radiotherapy treatment, which provides a new strategy for distinguishing the types of cancer cells and evaluating the radiotherapeutic efficacy of cancer cells. Furthermore, the functional microelectrode is integrated into an implantable probe for the in situ detection of surgically resected human specimens to distinguish cancer tissues from normal tissues. These will be of vital significance for cancer diagnoses and therapy in clinical practice. High-order 3D nanotube arrays (NTAs) arranged from mesoporous 2D N, B, P co-doped carbon network (NBP-CNW) were synthesize by a facile, well-controllable, eco-friendly and sustainable strategy using task-specific ionic liquids as precursors. NBP-CNW-NTAs modified flexible microelectrode was embed in microfluidic electrochemical biosensor chip for real-time tracking H2O2 secreted from different cancer cells (i.e., breast cancer cells, hepatoma cells and cervical cancer cells) with or without radiotherapy treatment. Furthermore, the functional microelectrode has been integrated into an implantable probe for in situ minimally invasive detection of surgically resected human breast specimens to identify the tumor tissues from the normal one. [ABSTRACT FROM AUTHOR]

The two-electron cytoplasmic reductase NAD(P)H:quinone oxidoreductase 1 is expressed in many tissues. NAD(P)H:quinone oxidoreductase 1 is well-known for being highly expressed in most cancers. Therefore, it could be a target for cancer therapy. Because it is a quinone reductase, many bioimaging probes based on quinone structures target NAD(P)H:quinone oxidoreductase 1 to diagnose tumours. Its expression is higher in tumours than in normal tissues, and using target drugs such as β-lapachone to reduce side effects in normal tissues can help. However, the physicochemical properties of β-lapachone limit its application. The problem can be solved by using nanosystems to deliver β-lapachone. This mini-review summarizes quinone-based fluorescent, near-infrared and two-photon fluorescent probes, as well as nanosystems for delivering the NAD(P)H:quinone oxidoreductase 1-activating drug β-lapachone. This review provides valuable information for the future development of probes and nano-delivery systems that target NAD(P)H:quinone oxidoreductase 1. [ABSTRACT FROM AUTHOR]

Objective: To verify the efficacy of Quxie Capsule in patients with metastatic colorectal cancer (mCRC). Methods: It was a block randomized, double-blind, placebo-controlled trial. Sixty patients with mCRC were randomized into 2 groups at a 1:1 ratio. The patients in the treatment group received conventional therapy including chemotherapy, radiotherapy, targeted therapy and supportive care, and Chinese herbal medicine combined with Quxie Capsule (each capsule of 0.4 g was orally administered at 50 mg/kg, twice daily, day 1–20, in a 30-day course) for 3 months. The patients in the control group received conventional therapy and Chinese herbal medicine combined with placebo for 3 months. Main outcome measures were overall survival (OS) and progression-free survival (PFS). Subgroup analysis was performed according to therapy lines, right or left-sided colon, targeted therapy and RAS gene status to determine the differences in PFS and OS between the two groups. Patients were followed up every 3 months until December 31st, 2018. Results: Median follow-up time was 19.4 months. The median OS was 23.9 months in the treatment group [95% confidence interval (CI) 15.9–28.5] vs. 14.3 months in the control group (95% CI 11.3–21.4, P<;0.05). Hazard ratio (95% CI) was 0.55 (0.31–0.95, P=0.040). There were no significant differences between the two groups in PFS (P>0.05). In the subgroups of ⩾second-line therapy, non-targeted therapy, RAS gene wild type and left-sided colon, the treatment group showed a significant survival benefit compared with the control group (P<;0.05 or P<;0.01), respectively. Conclusion: Quxie Capsule can reduce the risk of death and prolong the OS of patients with mCRC. (Registration No. ChiCTR-IOR-16009733) [ABSTRACT FROM AUTHOR]

Objectives: Several studies have compared the efficacy of hepatic resection (HR) and thermal ablation (TA) for unresectable tumors; however, results remain inconsistent. Most cohorts in previous studies were heterogeneous groups of synchronous colorectal liver metastases (CRLM) and extrahepatic metastases. This retrospective study aimed to compare the therapeutic efficacy between TA and HR in synchronous CRLM without extrahepatic metastases.Methods: Cases with initially synchronous CRLM without extrahepatic metastases between January 2007 and December 2018 were enrolled. Of the 448 cases, 346 received HR and 102 TA. Propensity score matching with a 1:1 ratio was used to improve the comparability between the HR and TA groups. Technical success, complications, disease-free survival (DFS), and overall survival (OS) were compared before and after matching.Results: All patients achieved technical success. Major complication rates in the HR and TA groups were, respectively, 36.7% and 8.8% (p < 0.001). Before matching, the 5-year OS and DFS (p = 0.004 and p = 0.020, respectively) were significantly higher in the HR group than in the TA group. After matching, no significant difference in the 5-year OS and DFS was found between the groups (p = 0.770 and p = 0.939, respectively). Local tumor progression rate was significantly higher in the TA group both before (p = 0.027) and after (p = 0.029) matching.Conclusions: For patients with CRC with synchronous CRLM, TA and HR provide comparable OS and DFS. TA is preferable if complete ablation is predicted.Key Points: • Thermal ablation and hepatic resection provide comparable overall survival and disease-free survival. • Thermal ablation is a safe and effective treatment for patients with colorectal cancer with synchronous liver metastases and has a lower major complication rate and higher repeatability than hepatic resection. • Thermal ablation is preferable if complete ablation is predicted. [ABSTRACT FROM AUTHOR]

Background: Although many reports have shown that preoperative dementia affects surgical prognosis, it is unclear whether the dementia based on Mini-Mental State Examination (MMSE) affect hospital mortality in old old patients undergoing elective gastrointestinal surgery. Aims: This study aims to investigate whether preoperative dementia might affect the outcomes of old old patients undergoing elective gastrointestinal surgery by evaluating with the MMSE. Methods: All patients aged ≥ 75 years who undergoing elective gastrointestinal surgery form January 2015 to December 2021 in a Chinese tertiary hospital were retrospectively analyzed. Their preoperative cognitive status was evaluated using the MMSE, and analysis was performed to compare the patients with MMSE score < 24 (dementia group) or MMSE score 24–30 (non-dementia group). Risk factors for hospital mortality were explored using multivariate logistic regression analysis. Results: 980 patients were rolled in the study, and 102 (10.4%) patients were in the dementia group. ICU and hospital stay were longer in the dementia group. Regarding the postoperative complications, the incidence of cerebral infarction (P = 0.014), delirium (P = 0.019), and pulmonary infection (P = 0.017) was more frequent in dementia group. Hospital mortality was 11% in the dementia group and 3% in the non-dementia group (P = 0.009). Multivariate logistic regression analysis revealed that dementia (P = 0.0135), preoperative lower albumin (P = 0.0018) and malignancy (P = 0.0212) were independent risk factors for hospital mortality. Conclusions: Among old old patients undergoing elective gastrointestinal surgery, hospital mortality was increased significantly for patients with dementia evaluating with the MMSE. [ABSTRACT FROM AUTHOR]

Applying discrete Calderón's identity, we study weighted multi-parameter mixed Hardy space H mix p (ω , ℝ n 1 × ℝ n 2 ) . Different from classical multi-parameter Hardy space, this space has characteristics of local Hardy space and Hardy space in different directions, respectively. As applications, we discuss the boundedness on H mix p (ω , ℝ n 1 × ℝ n 2 ) of operators in mixed Journé's class. [ABSTRACT FROM AUTHOR]

Ischemic stroke caused by intracranial vascular occlusion has become increasingly prevalent with considerable mortality and disability, which gravely burdens the global economy. Current relatively effective clinical treatments are limited to intravenous alteplase and thrombectomy. Even so, patients still benefit little due to the short therapeutic window and the risk of ischemia/reperfusion injury. It is therefore urgent to figure out the neuronal death mechanisms following ischemic stroke in order to develop new neuroprotective strategies. Regarding the pathogenesis, multiple pathological events trigger the activation of cell death pathways. Particular attention should be devoted to excitotoxicity, oxidative stress, and inflammatory responses. Thus, in this article, we first review the principal mechanisms underlying neuronal death mediated by these significant events, such as intrinsic and extrinsic apoptosis, ferroptosis, parthanatos, pyroptosis, necroptosis, and autophagic cell death. Then, we further discuss the possibility of interventions targeting these pathological events and summarize the present pharmacological achievements. [ABSTRACT FROM AUTHOR]

Low molecular weight proteins (LMWPs) in the bloodstream participate in various biological processes and are closely associated with disease status, whereas identification of serous LMWPs remains a great technical challenge due to the wide dynamic range of protein components. In this study, we constructed an integrated LMWP library by combining the LMWPs obtained by three enrichment methods (50% ACN, 20% ACN + 20 mM ABC, and 30 kDa) and their fractions identified by the data-dependent acquisition method. With this newly constructed library, we comprehensively profiled LMWPs in serum using data-independent acquisition and reliably achieved quantitative results for 75% serous LMWPs. When applying this strategy to quantify LMWPs in human serum samples, we could identify 405 proteins on average per sample, of which 136 proteins were with a MW less than 30 kDa and 293 proteins were with a MW less than 65 kDa. Of note, pre- and post-operative gastric carcinoma (GC) patients showed differentially expressed serous LWMPs, which was also different from the pattern of LWMP expression in healthy controls. In conclusion, our results showed that LMWPs could efficiently distinguish GC patients from healthy controls as well as between pre- and post-operative statuses, and more importantly, our newly developed LMWP profiling platform could be used to discover candidate LMWP biomarkers for disease diagnosis and status monitoring. [ABSTRACT FROM AUTHOR]

Amorphous calcium carbonate plays a key role as transient precursor in the early stages of biogenic calcium carbonate formation in nature. However, due to its instability in aqueous solution, there is still rare success to utilize amorphous calcium carbonate in biomedicine. Here, we report the mutual effect between paramagnetic gadolinium ions and amorphous calcium carbonate, resulting in ultrafine paramagnetic amorphous carbonate nanoclusters in the presence of both gadolinium occluded highly hydrated carbonate-like environment and poly(acrylic acid). Gadolinium is confirmed to enhance the water content in amorphous calcium carbonate, and the high water content of amorphous carbonate nanoclusters contributes to the much enhanced magnetic resonance imaging contrast efficiency compared with commercially available gadolinium-based contrast agents. Furthermore, the enhanced T1 weighted magnetic resonance imaging performance and biocompatibility of amorphous carbonate nanoclusters are further evaluated in various animals including rat, rabbit and beagle dog, in combination with promising safety in vivo. Overall, exceptionally facile mass-productive amorphous carbonate nanoclusters exhibit superb imaging performance and impressive stability, which provides a promising strategy to design magnetic resonance contrast agent. Sensitive, biocompatible and stable contrast agents for MRI are in demand. Here, the authors combine gadolinium ions with amorphous calcium carbonate to make stable paramagnetic amorphous carbonate nanoclusters with high MRI contrast and significantly improved biocompatibility over commercial gadolinium-based agents. [ABSTRACT FROM AUTHOR]

In this work, the "functionalization-polymerization" (FP) method has been used to construct fullerene-contained double-cable conjugated polymers with "donor-acceptor" backbones. It was realized via synthesizing a fullerene-contained monomer and performing Stille polymerization. With this method, a series of double-cable conjugated polymers with different fullerene contents were developed and applied into single-component organic solar cells. The power conversion efficiencies (PCEs) based on these polymers increased from 0.71% to 1.71% with the enhanced fullerene contents. The relatively low PCEs might be originated from the poor microstructure in these polymers. These new conjugated polymers with molecular heterojunction would show potential application in organic electronic devices. [ABSTRACT FROM AUTHOR]

Introduction: Glycerol is a byproduct from the biodiesel industry that can be biotransformed by Escherichia coli to high added-value products such as succinate under aerobic conditions. The main genetic engineering strategies to achieve this aim involve the mutation of succinate dehydrogenase (sdhA) gene and also those responsible for acetate synthesis including acetate kinase, phosphate acetyl transferase and pyruvate oxidase encoded by ackA, pta and pox genes respectively in the ΔsdhAΔack-ptaΔpox (M4) mutant. Other genetic manipulations to rewire the metabolism toward succinate consist on the activation of the glyoxylate shunt or blockage the pentose phosphate pathway (PPP) by deletion of isocitrate lyase repressor (iclR) or gluconate dehydrogenase (gnd) genes on M4-ΔiclR and M4-Δgnd mutants respectively. Objective: To deeply understand the effect of the blocking of the pentose phosphate pathway (PPP) or the activation of the glyoxylate shunt, metabolite profiles were analyzed on M4-Δgnd, M4-ΔiclR and M4 mutants. Methods: Metabolomics was performed by FT-IR and GC–MS for metabolite fingerprinting and HPLC for quantification of succinate and glycerol. Results: Most of the 65 identified metabolites showed lower relative levels in the M4-ΔiclR and M4-Δgnd mutants than those of the M4. However, fructose 1,6-biphosphate, trehalose, isovaleric acid and mannitol relative concentrations were increased in M4-ΔiclR and M4-Δgnd mutants. To further improve succinate production, the synthesis of mannitol was suppressed by deletion of mannitol dehydrogenase (mtlD) on M4-ΔgndΔmtlD mutant that increase ~ 20% respect to M4-Δgnd. Conclusion: Metabolomics can serve as a holistic tool to identify bottlenecks in metabolic pathways by a non-rational design. Genetic manipulation to release these restrictions could increase the production of succinate. [ABSTRACT FROM AUTHOR]

Protein therapeutics frequently face major challenges, including complicated production, instability, poor solubility, and aggregation. De novo protein design can readily address these challenges. Here, we demonstrate the utility of a topological refactoring strategy to design novel granulopoietic proteins starting from the granulocyte-colony stimulating factor (G-CSF) structure. We change a protein fold by rearranging the sequence and optimising it towards the new fold. Testing four designs, we obtain two that possess nanomolar activity, the most active of which is highly thermostable and protease-resistant, and matches its designed structure to atomic accuracy. While the designs possess starkly different sequence and structure from the native G-CSF, they show specific activity in differentiating primary human haematopoietic stem cells into mature neutrophils. The designs also show significant and specific activity in vivo. Our topological refactoring approach is largely independent of sequence or structural context, and is therefore applicable to a wide range of protein targets. Skokowa et al. reconstruct the fold of a granulopoietic cytokine, resulting in de novo, hyperstable, highly active proteins with therapeutic potential for treating several neutropenia disorders. [ABSTRACT FROM AUTHOR]