Engineering Escherichia coli to Efficiently Produce Colanic Acid with Low Molecular Mass and Viscosity.

Colanic acid (CA) is exopolysaccharide that presents growing potential in the food and healthcare industry as a versatile polymer. Previously, we have constructed the Escherichia coli strain WWM16 which can efficiently produce CA. In this study, WWM16 has been further engineered to produce a higher yield of CA with low molecular mass and viscosity. The gene mcbR encoding a transcriptional factor, and the genes opgD, opgG, and opgH related to the biosynthesis of osmoregulated periplasmic glucans were deleted in E. coli WWM16, and the resulting strain WWM166 produced 18.1 g/L CA. The expression level of wcaD encoding the polymerase in WWM166 was downregulated using CRISPRi. As a result, the strain WWM166/pWpD1 could produce 49.9 g/L CA with lower molecular mass. CA products were purified from both WWM166 and WWM166/pWpD1, and their molecular mass, viscosity, fluidity, hygroscopicity, and antioxidant activity were determined and compared. These findings demonstrate the potential application of CA with different molecular masses to prolong life and protect skin in the food and cosmetic industries.

Treatment outcomes and prognostic factors of the apical barrier technique with premixed calcium silicate-based putty in necrotic permanent teeth with open apices: a retrospective cohort study with up to six years follow-up.

OBJECTIVES: To evaluate treatment outcomes of the apical barrier technique with premixed calcium silicate-based putty for treating necrotic permanent teeth with open apices and to identify prognostic factors. MATERIALS AND METHODS: Permanent teeth with necrotic pulps and open apices treated by the apical barrier technique with premixed calcium silicate-based putty, with a minimum follow-up of 12 months, were included. Treatment outcomes were based on clinical signs, symptoms, and radiographic evaluation. The treatment outcome was dichotomized into success or failure according to strict and loose criteria. The chi-square test (or Fisher's exact test) and multiple logistic regression analysis were used to evaluate possible prognostic factors associated with treatment outcomes. RESULTS: Seventy-four teeth with a follow-up time of 12-72 months (mean, 25.74 ± 14.36 months) were included in the final evaluation. The success rate was 97.30% using the loose criteria and 66.22% using the strict criteria. Multiple logistic regression analysis indicated that the size of pre-operative periapical lesion (≥ 5 mm) (odds ratio [OR]: 18.96; P = 0.0153) and root canal underfilling (OR: 8.341; P = 0.0448) were significant predictors for treatment failure under the strict criteria. CONCLUSION: The apical barrier technique with premixed calcium silicate-based putty is a highly successful procedure for treating necrotic permanent teeth with open apices after an observation period of up to 6 years. Treatment success under the strict criteria is primarily affected by the size of the pre-operative periapical lesion and the apical extent of root-filling. CLINICAL RELEVANCE: Careful case selection and ensuring adequate root filling quality are essential to the successful outcome of the apical barrier technique with premixed calcium silicate-based putty.

Lacustrine carbon sink: A hidden driver of the Late Cretaceous Cooling Event.

Lacustrine systems since the Mesozoic have sequestered large quantities of organic carbon, which may have important value for global climate cooling, but there is still a lack of geological evidence of this sequestration. Taking the Songliao Basin in China as a case study, we elucidate the important function of lacustrine basins as sinks of a large amount of organic carbon, particularly when the contemporaneous marine sediments were poor sinks of organic carbon. Volcanic activities and orbital forcing were likely key factors influencing the water transportation between the land and oceans, as well as the alternating burial of organic carbon in the oceans and land. Microorganisms related to methane metabolism may have been highly involved in the mineralization and sequestration of lacustrine organic carbon. This study provides new insights into the coupled carbon-water cycle between the land and oceans and the influence of this process on global climate evolution.

Impact and migration behavior of triclosan on waste-activated sludge anaerobic digestion.

Triclosan (TCS), a hydrophobic antibacterial agent, is extensive application in daily life. Despite a low biodegradability rate, its hydrophobicity results in its accumulation in waste-activated sludge (WAS) during domestic and industrial wastewater treatment. While anaerobic digestion is the foremost strategy for WAS treatment, limited research has explored the interphase migration behavior and impacts of TCS on WAS degradation during anaerobic digestion. This study revealed TCS migration between solid- and liquid-phase in WAS digestion. The solid-liquid distribution coefficients of TCS were negative for proteins and polysaccharides and positive for ammonium. High TCS levels promoted volatile-fatty-acid accumulation and reduced methane production. Enzyme activity tests and functional prediction indicated that TCS increased enzyme activity associated with acid production, in contrast to the inhibition of key methanogenic enzymes. The findings of the TCS migration behavior and its impacts on WAS anaerobic digestion provide an in-depth understanding of the evolution of enhanced TCS-removing technology.

Advancements in Iron Oxide Nanoparticles for Multimodal Imaging and Tumor Theranostics.

The emergence of nanomedicine offers renewed promise in the diagnosis and treatment of diseases. Due to their unique physical and chemical properties, iron oxide nanoparticles (IONPs) exhibit widespread application in the diagnosis and treatment of various ailments, particularly tumors. IONPs have magnetic resonance (MR) T1/T2 imaging capabilities due to their different sizes. In addition, IONPs also have biocatalytic activity (nanozymes) and magnetocaloric effects. They are widely used in chemodynamic therapy (CDT), magnetic hyperthermia treatment (MHT), photodynamic therapy (PDT), and drug delivery. This review outlines the synthesis, modification, and biomedical applications of IONPs, emphasizing their role in enhancing diagnostic imaging (including single-mode and multimodal imaging) and their potential in cancer therapies (including chemotherapy, radiotherapy, CDT, and PDT). Furthermore, we briefly explore the challenges in the clinical application of IONPs, such as surface modification and protein adsorption, and put forward opinions on the clinical transformation of IONPs.

Synthetic routes and clinical application of Small-Molecule HER2 inhibitors for cancer therapy.

This comprehensive review undertakes a meticulous scrutiny of the synthesis and clinical applications pertaining to small-molecule tyrosine kinase inhibitors (TKIs) directed towards the human epidermal growth factor receptor 2 (HER2), a pivotal protagonist in the pathogenesis of cancer. Focused on compounds like lapatinib, neratinib, and tucatinib, the review delves into the intricate synthesis strategies, emphasizing the challenges associated with their structural complexity. The clinical utilization of HER2 TKIs underscores noteworthy strides in the therapeutic landscape for HER2-positive breast and gastric malignancies. Lapatinib, a dual HER2/ epidermal growth factor receptor (EGFR) inhibitor, has demonstrated efficacy in combination therapies, addressing the need for overcoming resistance mechanisms. Neratinib, an irreversible HER2 inhibitor, presents a promising avenue for patients with refractory tumors. Tucatinib, strategically engineered to traverse the blood-brain barrier, epitomizes a groundbreaking advancement in the management of metastatic HER2-positive breast cancer manifesting cerebral involvement. Despite their success, challenges such as resistance mechanisms and off-target effects persist, urging continuous research for the development of next-generation HER2 TKIs. This comprehensive review serves as a valuable resource for pharmaceutical scientists, offering insights into the synthetic intricacies and clinical impact of small-molecule TKIs targeting HER2.

Combining binding pocket mutagenesis and substrate tunnel engineering to improve an (R)-selective transaminase for the efficient synthesis of (R)-3-aminobutanol.

(R)-selective transaminases have the potential to act as efficient biocatalysts for the synthesis of important pharmaceutical intermediates. However, their low catalytic efficiency and unfavorable equilibrium limit their industrial application. Seven (R)-selective transaminases were identified using homologous sequence mining. Beginning with the optimal candidate from Mycolicibacterium hippocampi, virtual mutagenesis and substrate tunnel engineering were performed to improve catalytic efficiency. The obtained variant, T282S/Q137E, exhibited 3.68-fold greater catalytic efficiency (kcat/Km) than the wild-type enzyme. Using substrate fed-batch and air sweeping processes, effective conversion of 100 mM 4-hydroxy-2-butanone was achieved with a conversion rate of 93 % and an ee value > 99.9 %. This study provides a basis for mutation of (R)-selective transaminases and offers an efficient biocatalytic process for the asymmetric synthesis of (R)-3-aminobutanol.

Farmland's silent threat: Comprehensive multimedia assessment of micropollutants through non-targeted screening and targeted analysis in agricultural systems.

Intricate agricultural ecosystems markedly influence the dynamics of organic micropollutants, posing substantial threats to aquatic organisms and human health. This study examined the occurrence and distribution of organic micropollutants across soils, ditch sediment, and water within highly intensified farming setups. Using a non-targeted screening method, we identified 405 micropollutants across 10 sampling sites, which mainly included pesticides, pharmaceuticals, industrial chemicals, and personal care products. This inventory comprised emerging contaminants, banned pesticides, and controlled pharmaceuticals that had eluded detection via conventional monitoring. Targeted analysis showed concentrations of 3.99-1021 ng/g in soils, 4.67-2488 ng/g in sediment, and 12.5-9373 ng/L in water, respectively, for Σ40pesticides, Σ8pharmaceuticals, and Σ3industrial chemicals, indicating notable spatial variability. Soil organic carbon content and wastewater discharge were likely responsible for their spatial distribution. Principal component analysis and correlation analysis revealed a potential transfer of micropollutants across the three media. Particularly, a heightened correlation was decerned between soil and sediment micropollutant levels, highlighting the role of sorption processes. Risk quotients surpassed the threshold of 1 for 13-23 micropollutants across the three media, indicating high environmental risks. This study highlights the importance of employing non-targeted and targeted screening in assessing and managing environmental risks associated with micropollutants.

Atypical Guillain-Barré syndrome with positive anti-sulfatide, anti-GT1b, and anti-GT1a antibodies: A case report.

BACKGROUND: The role of diverse antibodies in mediating peripheral nerve injury in Guillain-Barré syndrome (GBS) is becoming clearer, but positivity for multiple antibodies in one case is uncommon. To our knowledge, this is the first case involving GBS with positive anti-sulfatide, anti-GT1a, and anti-GT1b antibodies. CASE SUMMARY: A 20-year-old female patient was admitted to the hospital due to weakness of limbs for 5 d, and deterioration of the weakness and muscle aches for 1 d. The patient's limbs were weak, but the tendon reflexes in the part of the limbs were normal. There was no comorbid peripheral nociception or deep sensory dysfunction. She was diagnosed with GBS and was discharged after receiving intravenous human immunoglobulin pulse therapy. CONCLUSION: In this article, the clinical manifestations, neurophysiological examination, and auxiliary examination findings of a GBS patient positive for multiple antibodies were analyzed to improve the identification of the disease by clinical physicians at an early stage.

Endodontic Treatment of a Mandibular Second Molar Featuring Vertucci Type V Configuration in the Distal Root: A Case Report.

Successful root canal treatment relies primarily on thorough shaping, cleaning, and filling of the entire root canal system. Neglecting even a single canal can significantly raise the risk of post-treatment apical periodontitis. While the distal root of mandibular second molars typically has one canal, they can also present with anatomical variations, including the presence of a Vertucci Type V configuration. This article discusses a case in which a Vertucci Type V configuration in a mandibular second molar was effectively identified and treated.

Influence of ear tags on the results of body composition analysis in mice.

BACKGROUND: The aim of this study was to investigate the influence of marking methods on the outcomes of body composition analysis and provide guidance for the selection of marking methods in mouse body composition analysis. METHODS: Male C57BL/6J mice aged 6 weeks were randomly assigned for pre- and post- ear tagging measurements. The body composition of the mice was measured using a small animal body composition analyzer, which provided measurements of the mass of fat, lean, and free fluid. Then, the mass of fat, lean and free fluid to body weight ratio was gained. Further data analysis was conducted to obtain the range and coefficient of variation in body composition measurements for each mouse. The distribution of fat and lean tissue in the mice was also analyzed by comparing the fat-to-lean ratio. RESULTS: (1) The mass of all body composition components in the ear tagging group was significantly lower than that in the control group. (2) There was a significant increase in the range and coefficient of variation of body composition measurements between the ear tagging group and the control group. (3) The fat-to-lean ratio in the ear tagging group was significantly lower than that in the control group. CONCLUSIONS: Ear tagging significantly lowered the results of body composition analysis in mice and higher the results of measurement error. Therefore, ear tagging should be avoided as much as possible when conducting body composition analysis experiments in mice.

Preparation and Characterization of Prickly Ash Peel Oleoresin Microcapsules and Flavor Retention Analysis.

Prickly ash peel oleoresin (PPO) is a highly concentrated oil of Prickly ash essential oil and has a stronger aroma. However, its low water solubility, high volatility, difficulty in transport and storage, and decomposition by light, heat, and oxygen limit its wider application. To solve this problem, this study used freeze-drying or spray-drying, with soybean protein isolate (SPI) or gum Arabic (GA), combined with aqueous maltodextrin (MD) as the encapsulating agents to prepare four types of PPO microcapsules (POMs). Spray-dried microcapsules with GA as the encapsulating agent achieved a high encapsulation efficiency (EE) of 92.31 ± 0.31%, improved the thermal stability of the PPO, and had spherical morphology. (Headspace solid-phase microextraction/gas chromatography-mass spectrometry) HS-SPME/GC-MS detected 41 volatile compounds in PPO; of these, linalool, ß-myrcene, sabinene, and D-limonene were identified as key flavor components. Principal component analysis (PCA) effectively distinguished the significant differences in flavor between PPO, spray-dried SPI/MD microcapsules (SS), and spray-dried GA/MD microcapsules (SG). During 15 days of air-exposure, the loss of flavor from SG (54.62 ± 0.54%) was significantly lower than PPO (79.45 ± 1.45%) and SS (57.55 ± 0.36%). During the air-exposure period, SG consistently had the highest antioxidant capacity, making it desirable for PPO packaging, and expanding its potential applications within the food industry.

Procalcitonin and C-reactive protein as early predictors in patients at high risk of colorectal anastomotic leakage.

OBJECTIVE: To assess the diagnostic value of C-reactive protein (CRP) and procalcitonin (PCT) for anastomotic leakage (AL) following colorectal surgery. METHODS: We retrospectively analyzed data for patients who underwent colorectal surgery at our hospital between November 2019 and December 2023. CRP and PCT were measured postoperatively to compare patients with/without AL, and changes were compared between low- and high-risk groups. Receiver operating characteristic (ROC) curve analysis was used to assess the diagnostic accuracy of CRP and PCT to identify AL in high-risk patients. RESULTS: Mean CRP was 142.53 mg/L and 189.57 mg/L in the low- and high-risk groups, respectively, on postoperative day (POD)3. On POD2, mean PCT was 2.75 ng/mL and 8.16 ng/mL in low- and high-risk patients, respectively; values on POD3 were 3.53 ng/mL and 14.86 ng/mL, respectively. The areas under the curve (AUC) for CRP and PCT on POD3 were 0.71 and 0.78, respectively (CRP cut-off: 235.64 mg/L; sensitivity: 96%; specificity: 89.42% vs PCT cut-off: 3.94 ng/mL; sensitivity: 86%; specificity: 93.56%; AUC: 0.78). The AUC, sensitivity, and specificity for the combined diagnostic ability of CRP and PCT on POD3 were 0.92, 90%, and 100%, respectively (cut-off: 0.44). CONCLUSIONS: Combining PCT and CRP on POD3 enhances the diagnostic accuracy for AL.

Endodontic Treatment of a Dilacerated Maxillary Second Premolar With a Severely Curved Root Canal: A Case Report and Literature Review.

Dilaceration is a developmental anomaly characterized by a sharp change in the axial inclination between the crown and the root of a tooth. Severe root curvature in a dilacerated tooth can greatly complicate root canal treatment. This case report details the successful endodontic treatment of a dilacerated maxillary second premolar with significant root curvature. It highlights the importance of a thorough understanding of root canal anatomy and demonstrates the effectiveness of using pre-curved hand files along with heat-treated nickel-titanium rotary instruments in navigating complex root structures to achieve successful treatment outcomes.

Antimicrobial efficacy of DJK-5 peptide in combination with EDTA against biofilms in dentinal tubules: Primary irrigation, recovery and re-irrigation.

AIM: To investigate the dynamic recovery of biofilms within dentinal tubules after primary irrigation with different protocols, and to evaluate the efficacy of various re-irrigation protocols on recovered biofilm, considering factors such as smear layer, nutrient conditions, and primary irrigants. METHODOLOGY: A total of 416 mono or multi-species biofilms samples were prepared from human teeth and incubated for 3 weeks. After inducing a smear layer on half of the samples, all specimens were irrigated with one of the following irrigant sequences: (1) 6% NaOCl +17% EDTA, (2) 6% NaOCl +8.5% EDTA, (3) 6% NaOCl and (8.5% EDTA +10 µg/mL DJK-5 antimicrobial peptide), or (4) sterile water. Thirty-two samples were used to assess immediate effect, whilst the rest were re-incubated to assess biofilms recovery. Nutrient conditions were defined based on whether culture media were changed (nutrient-rich) or not (nutrient-poor) during re-incubation. After 16 weeks, recovered biofilms underwent re-irrigation using four additional protocols, with or without DJK-5 peptide, based on primary irrigants. Confocal laser scanning microscopy was employed to evaluate immediate irrigant effects, biofilms recovery intervals (1, 3, 5, 8, 12, and 16 weeks after primary irrigation), and re-irrigation effects at the 16-week. Statistical analysis included one-way anova and two-way mixed anova tests. RESULTS: The DJK-5 peptide irrigation protocols demonstrated the highest killing rates during primary irrigation and resulted in a longer biofilms recovery time of 16 weeks compared to non-peptide protocols (p < .001). Both primary irrigation type and smear layer presence significantly influenced biofilms recovery (p < .001). In the absence of smear layer, re-irrigation efficacy didn't significantly differ from primary irrigation, regardless of primary irrigation type or nutrient conditions. However, with a smear layer present, re-irrigation led to significantly higher proportion of dead bacteria compared to primary irrigation (p < .05). Inclusion of the DJK-5 peptide into the re-irrigation protocol displayed superior killing rate compared to other protocols (p < .001). CONCLUSIONS: Biofilms exhibited susceptibility to both peptide and non-peptide protocols during re-irrigation, irrespective of nutrient conditions or primary irrigation protocols. The DJK-5 peptide irrigation protocols consistently displayed superior effectiveness compared to non-peptide protocols.

One-Pot Two-Stage Biocatalytic Cascade to Produce l-Phosphinothricin by Two Enantioselective Complementary Aminotransferases at High Substrate Loading via a Deracemization Process.

The bioderacemization of racemic phosphinothricin (D, L-PPT) is a promising route for the synthesis of l-phosphinothricin (L-PPT). However, the low activity and tolerance of wild-type enzymes restrict their industrial applications. Two stereocomplementary aminotransferases with high activity and substrate tolerance were identified in a metagenomic library, and a one-pot, two-stage artificial cascade biocatalytic system was developed to produce L-PPT through kinetic resolution and asymmetric amination. We observed that 500 mM D, L-PPT (100 g/L) could be converted into L-PPT with 94% final conversion and >99.9% enantiomeric excess (e.e.) within 24 h, with only 0.02 eq amino acceptor pyruvate and 1.2 eq amino donor l-aspartate required. The process could be scaled up to 10 L under sufficient oxygen and stirring. The superior catalytic performance of this system provides an eco-friendly and sustainable approach to the industrial deracemization of D, L-PPT to L-PPT.

Midbrain glutamatergic circuit mechanism of resilience to socially transferred allodynia in male mice.

The potential brain mechanism underlying resilience to socially transferred allodynia remains unknown. Here, we utilize a well-established socially transferred allodynia paradigm to segregate male mice into pain-susceptible and pain-resilient subgroups. Brain screening results show that ventral tegmental area glutamatergic neurons are selectively activated in pain-resilient mice as compared to control and pain-susceptible mice. Chemogenetic manipulations demonstrate that activation and inhibition of ventral tegmental area glutamatergic neurons bi-directionally regulate resilience to socially transferred allodynia. Moreover, ventral tegmental area glutamatergic neurons that project specifically to the nucleus accumbens shell and lateral habenula regulate the development and maintenance of the pain-resilient phenotype, respectively. Together, we establish an approach to explore individual variations in pain response and identify ventral tegmental area glutamatergic neurons and related downstream circuits as critical targets for resilience to socially transferred allodynia and the development of conceptually innovative analgesics.

Enhancing frozen dough quality: Investigating the impact of soy hull polysaccharide (SHP) on rheological properties and microstructure.

The effects of SHP on the texture, rheological properties, starch crystallinity and microstructure of frozen dough were investigated. The efficacy of SHP in enhancing dough quality is concentration-dependent, with frozen dough containing 1.5% SHP exhibiting hardness comparable to fresh dough without SHP (221.31 vs. 221.42 g). Even at 0.5% SHP, there is a noticeable improvement in frozen dough quality. The rheological results showed that the viscoelasticity of dough increased with higher SHP concentration. What's more, XRD and SEM results indicated that the SHP's hydrophilicity reduces the degree of starch hydrolysis, slows down the damage of starch particles during freezing, and consequently lowers the crystallinity of starch. Additionally, CLSM observations revealed that SHP enhances the gluten network structure, diminishing the appearance of holes. Therefore, the physical, chemical properties, and microstructure of frozen dough with SHP demonstrate significant enhancement, suggesting SHP's promising antifreeze properties and potential as a food antifreeze agent.

An investigation of a strengthening polysaccharide interfacial membrane strategy utilizing an anionic polysaccharide-alkaline ligand interfacial assembly for all-liquid printing.

The applications of polysaccharides as emulsifiers are limited due to the lack of hydrophobicity. However, traditional hydrophobic modification methods used for polysaccharides are complicated and involve significant mechanical and thermal losses. In this study, soy hull polysaccharide (SHP) and terminally aminopropylated polydimethylsiloxane (NPN) were selected to investigate the feasibility of a simple and green interfacial membrane strengthening strategy based on the interfacial polymerization of anionic polysaccharides and fat-soluble alkaline ligands. Our results show that deprotonated SHP and protonated NPN can be complexed at the water/oil (W/O) interface, reduce interfacial tension, and form a strong membrane structure. Moreover, they can quickly form a membrane at the W/O interface upon the moment of contact to produce stable all-liquid printing products with complex patterns. However, the molecular weight of NPN affects the complexation reaction. Consequently, this study has long-term implications to expanding the areas of application for anionic polysaccharides.