Construction of an Escherichia coli Strain Lacking Fimbriae by Deleting 64 Genes and Its Application for Efficient Production of Poly(3-Hydroxybutyrate) and l-Threonine.

Escherichia coli contains 12 chaperone-usher operons for biosynthesis and assembly of various fimbriae. In this study, each of the 12 operons was deleted in E. coli MG1655, and the resulting 12 deletion mutants all grew better than the wild type, especially in the nutrient-deficient M9 medium. When the plasmid pBHR68 containing the key genes for polyhydroxyalkanoate production was introduced into these 12 mutants, each mutant synthesized more polyhydroxyalkanoate than the wild-type control. These results indicate that the fimbria removal in E. coli benefits cell growth and polyhydroxyalkanoate production. Therefore, all 12 chaperone-usher operons, including 64 genes, were deleted in MG1655, resulting in the fimbria-lacking strain WQM026. WQM026 grew better than MG1655, and no fimbria structures were observed on the surface of WQM026 cells. Transcriptomic analysis showed that in WQM026 cells, the genes related to glucose consumption, glycolysis, flagellar synthesis, and biosynthetic pathways of some key amino acids were upregulated, while the tricarboxylic acid cycle-related genes were downregulated. When pBHR68 was introduced into WQM026, huge amounts of poly-3-hydroxybutyrate were produced; when the plasmid pFW01-thrA*BC-rhtC, containing the key genes for l-threonine biosynthesis and transport, was transferred into WQM026, more l-threonine was synthesized than with the control. These results suggest that this fimbria-lacking E. coli WQM026 is a good host for efficient production of polyhydroxyalkanoate and l-threonine and has the potential to be developed into a valuable chassis microorganism. IMPORTANCE In this study, we investigated the interaction between the biosynthesis and assembly of fimbriae and intracellular metabolic networks in E. coli. We found that eliminating fimbriae could effectively improve the production of polyhydroxyalkanoate and l-threonine in E. coli MG1655. These results contribute to understanding the necessity of fimbriae and the advantages of fimbria removal for industrial microorganisms. The knowledge gathered from this study may be applied to the development of superior chassis microorganisms.

Recurrence-Related Factors of Medication-Related Osteonecrosis of the Jaw: A Five-Year Experience.

PURPOSE: The treatment of medication-related osteonecrosis of the jaw (MRONJ) is greatly challenging for surgeons. In this study, we reviewed patients with MRONJ treated in our hospital in the past 5 years and explored the risk factors of recurrence. METHODS: A retrospective cohort study was conducted to review the patients with MRONJ from January 1, 2016 to December 31, 2020. All patients received a monthly intravenous application of zoledronic acid. The primary outcome variable was the treatment outcome during follow-up. The predictor variables were local and systemic factors related to the treatment outcome. Multivariate logistic regression analysis was performed to identify the risk factors of recurrence after MRONJ treatment. RESULTS: A total of 58 patients (62 sites) were included in this study. In multivariate regression analysis, the risk factor associated with recurrence after MRONJ treatment was the duration of medication of more than 18 months (odds ratio = 7.346; 95% confidence interval: 1.461-36.946; P = .016). CONCLUSIONS: Using zoledronic acid over 18 months may increase the risk of recurrence in MRONJ patients.

Pollution characteristics and potential health effects of airborne microplastics and culturable microorganisms during urban haze in Harbin, China.

In this study, active sampling technology was used to collect microplastics (MPs) and microorganisms simultaneously on haze days in Harbin, China. Airborne MPs concentrations in Junior high school (162.4 ± 44.6 particles/m3) with high vehicular and pedestrian traffic was higher than those in University (63.2 ± 21.8 particles/m3) and Park (12.8 ± 5.5 particles/m3). More airborne MPs were detected in the night samples than in the morning and noon samples. The majority (69.06 %) of airborne MPs measured less than 100 µm, with fibers (69.4 %) being the predominant form. Polyesters and polyethylene were the dominant polymers. In addition, airborne MPs concentrations were positively correlated with microorganisms and PM10 concentrations, and the health hazards associated with microorganisms and MPs exposure via inhalation far exceeded those associated with skin contact, which can serve as a theoretical foundation for considering MPs as indicators of air quality in the future.

Correlation between facial attractiveness and facial components assessed by laypersons and orthodontists.

BACKGROUND/PURPOSE: Relationship between facial attractiveness and facial components is not completely clear. The aim was to analyze the correlation between facial attractiveness and facial components assessed by laypersons and orthodontists. MATERIALS AND METHODS: The attractiveness of 100 adult patients (50 males and 50 females with their frontal, lateral, and frontal smiling photos), before orthodontic treatment, was assessed by 24 laypersons and 24 orthodontists using visual analog scale. Pearson correlation coefficient and stepwise regression were used for statistical analysis. RESULTS: When laypersons assessed, a significant correlation was found between facial attractiveness and the chin (r = 0.671), eyes (r = 0.669), lips (r = 0.585), hair (r = 0.527), teeth (r = 0.338), and nose (r = 0.247); the chin was responsible for 45.1% of the variation in facial attractiveness, the eyes for 14.3%, and the lips for 0.8%. When orthodontists assessed, a significant correlation was found between facial attractiveness and the lips (r = 0.789), eyes (r = 0.646), hair (r = 0.613), chin (r = 0.565), nose (r = 0.264), and teeth (r = 0.221); the lips were responsible for 62.2% of the variation in facial attractiveness, the eyes for 8.2%, the chin for 1.4%, and the hair for 0.5%. CONCLUSION: When laypersons evaluate, the chin contributes the most to facial attractiveness, and when orthodontists evaluate, the lips contribute the most. Whether in the evaluation of laypersons or orthodontists, the contribution of teeth to facial attractiveness is significantly less than that of the lips and chin.

Anti-Glioma Effect with Targeting Therapy Using Folate Modified Nano-Micelles Delivery Curcumin.

Targeted drug delivery systems have currently demonstrated considerable potential clinical benefits in cancer treatment. Curcumin has become a candidate anti-tumor drug for the therapy of glioblastoma multiforme (GBM) by increasing cell apoptosis and suppressing cell proliferation. In current research, we explored a novel targeted drug delivery system with a self-assembly measure by curcumin, MPEG-PLA and Fa-PEG-PLA. Compared with free curcumin and Cur/MPEG-PLA, Cur/Fa-PEG-PLA can remarkably suppress the growth of GL261 cells and promote apoptotic rate. Moreover, after the procession of tumor-bearing mice with curcumin/Fa-PEG-PLA complex, tumor growth in subcutaneous and intracranial tumor models were repressed via suppressing angiogenesis and facilitating apoptosis in vivo. The Curcumin/Fa-PEG-PLA nanoparticle may be a novel drug for the therapy of GBM.

Phosphatidylinositol 3-Kinase/Akt signal pathway resists the apoptosis and inflammation in human extravillous trophoblasts induced by Porphyromonas gingivalis.

Epidemiological studies suggested that periodontitis is a risk factor for pregnancy complications including preterm birth. Porphyromonas gingivalis, a vital periodontal pathogen found in amniotic fluid and intact membranes of women who deliver preterm low birth weight infants, is thought to contribute to preterm labor. However, molecular and cellular interactions between P. gingivalis and placental cells are not clear. In this study, we investigated the effect of P. gingivalis on human extravillous trophoblasts and observed that it triggered apoptosis and inflammation and that Akt was activated in this process. In addition, when Akt activation was inhibited, apoptosis and inflammation was significantly increased. Thus, P. gingivalis infection contributes to preterm low birth weight infants by triggering excessive inflammation and increasing apoptosis in trophoblasts and that the Phosphatidylinositol 3-Kinase/Akt signaling pathway is involved in the regulation of Pg-induced apoptosis and inflammation.

Development of Epigallocatechin-3-gallate-Encapsulated Nanohydroxyapatite/Mesoporous Silica for Therapeutic Management of Dentin Surface.

In dental clinic, unsatisfactory management of the dentin surface after dentin exposure often leads to the occurrence of dentin hypersensitivity and caries. Current approaches can occlude the tubules on the dentin surface to relieve dentin hypersensitivity; however, the blocked tubules are generally weak in combating daily tooth erosion and abrasion. Moreover, cariogenic bacteria, such as Streptococcus mutans, produce biofilm on the dentin surface, causing caries and compromising the tubules' sealing efficacy. To overcome this problem, the present study focused on establishing a versatile biomaterial, epigallocatechin-3-gallate-encapsulated nanohydroxyapatite/mesoporous silica nanoparticle (EGCG@nHAp@MSN), for therapeutic management of the dentin surface. The effectiveness of the biomaterial on dentinal tubule occlusion, including resistances against acid and abrasion, was evaluated by field-emission scanning electron microscopy (FESEM) and dentin permeability measurement. The inhibitory capability of the biomaterial on S. mutans biofilm formation was investigated by confocal laser scanning microscopy (CLSM), 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, colony forming units (CFU) counts, and FESEM. Results demonstrated for the first time that the use of EGCG@nHAp@MSN on the dentin surface was capable of effectively occluding dentinal tubules, reducing dentin permeability, and achieving favorable acid- and abrasion-resistant stability. Furthermore, EGCG@nHAp@MSN held the capability to continuously release EGCG, Ca, and P, and significantly inhibit the formation and growth of S. mutans biofilm on the dentin surface. Thus, the development of EGCG@nHAp@MSN bridges the gap between multifunctional concept and dental clinical practice and is promising in providing dentists a therapeutic strategy for the management of the dentin surface to counter dentin hypersensitivity and caries.

Interferon-Gamma and Fas Are Involved in Porphyromonas gingivalis-Induced Apoptosis of Human Extravillous Trophoblast-Derived HTR8/SVneo Cells via Extracellular Signal-Regulated Kinase 1/2 Pathway.

BACKGROUND: A number of studies recently revealed a link between periodontal disease and preterm birth (PTB). PTB can be induced by dental infection with Porphyromonas gingivalis (Pg), a periodontopathic bacterium. This study aims to investigate responses of human extravillous trophoblast-derived HTR8/SVneo cells to Pg infection. METHODS: Cell apoptosis, cell viability, protein expression, and cytokine production in HTR8 cells were measured via: 1) flow cytometry, 2) CCK-8 assay, 3) western blot, and 4) enzyme-linked immunosorbent assay methods, respectively. RESULTS: Pg decreased cell viability and increased cell apoptosis, active caspase-3 and Fas expression, and interferon-gamma (IFN-γ) secretion in HTR8 cells. Extracellular signal-regulated kinase (ERK) 1/2 inhibitor U0126 and FasL neutralizing antibody NOK1 that blocks FasL/Fas interaction both significantly suppressed Pg-induced apoptosis. U0126 also inhibited IFN-γ secretion and Fas expression close to control levels. Moreover, treatment with recombinant IFN-γ also significantly decreased number of viable HTR8 cells and increased Fas expression, suggesting IFN-γ may play an important role in Pg-induced apoptosis of HTR8 cells, at least partially through regulation of Fas expression. CONCLUSIONS: To the best of the authors' knowledge, this is the first study to demonstrate Pg induces IFN-γ secretion, Fas expression, and apoptosis in human extravillous trophoblast-derived HTR8/SVneo cells in an ERK1/2-dependent manner, and IFN-γ (explored by recombinant IFN-γ) and Fas are involved in Pg-induced apoptosis. The finding that Pg infection abnormally regulates inflammation and apoptosis of human trophoblasts may give new insights into the possible link of PTB with maternal periodontal disease and periodontal pathogens.

Porphyromonas gingivalis induces IL-8 and IFN-gamma secretion and apoptosis in human extravillous trophoblast derived HTR8/SVneo cells via activation of ERK1/2 and p38 signaling pathways.

INTRODUCTION: Preterm birth is a major cause for infant mortality and morbidity. A large number of studies have suggested a link between periodontal disease and preterm birth. The purpose of this study was to investigate the interaction between a periodontopathic bacterium Porphyromonas gingivalis and human extravillous trophoblast derived HTR8/SVneo cells. METHODS: Production of cytokines in HTR8 cells was measured via ELISA. Annexin V/PI flow cytometry was performed to assess apoptosis. Protein expression was measured by western blot. Specific pharmacological inhibitors were used to inactivate relevant signaling pathways (p38 MAPK, SB203580; ERK1/2, U0126; JNK, SP600125; NF-κB, JSH-23) to determine their roles in inflammation and apoptosis. RESULTS: HTR8 cells released significant amounts of IL-8 and IFN-γ during exposure to P. gingivalis. Meanwhile, the percentages of both early and late apoptotic cells increased significantly in response to P. gingivalis. The most significant effect on inflammation was found using SB203580 and U0126, followed by SP600125 and JSH-23. Moreover, U0126 and SB203580 both partially but significantly suppressed P. gingivalis-induced apoptosis, with a large effect by U0126. Additionally, both heat-killed P. gingivalis and P. gingivalis lipopolysaccharide significantly induced IL-8 production. CONCLUSION: P. gingivalis induces inflammation and apoptosis in HTR8 cells, and we demonstrated for the first time that activation of ERK1/2 and p38 MAPK pathways participates in P. gingivalis-induced inflammation and apoptosis. The abnormal regulation of inflammation and apoptosis in human trophoblasts by P. gingivalis infection may give new insights into how maternal periodontal disease and periodontal pathogens might be linked to preterm birth.

Improved 4-chlorophenol dechlorination at biocathode in bioelectrochemical system using optimized modular cathode design with composite stainless steel and carbon-based materials.

This study developed and optimized a modular biocathode materials design in bioelectrochemical system (BES) using composite metal and carbon-based materials. The 4-chlorophenol (4-CP) dechlorination could be improved with such composite materials. Results showed that stainless steel basket (SSB) filled with graphite granules (GG) and carbon brush (CB) (SSB/GG/CB) was optimum for dechlorination, followed by SSB/CB and SSB/GG, with rate constant k of 0.0418 ± 0.0002, 0.0374 ± 0.0004, and 0.0239 ± 0.0002 h(-1), respectively. Electrochemical impedance spectroscopy (EIS) demonstrated that the composite materials with metal can benefit the electron transfer and decrease the charge transfer resistance to be 80.4 Ω in BES-SSB/GG/CB, much lower than that in BES-SSB (1674.3 Ω), BES-GG (387.3 Ω), and BES-CB (193.8 Ω). This modular cathode design would be scalable with successive modules for BES scale-up, and may offer useful information to guide the selection and design of BES materials towards dechlorination improvement in wastewater treatment.

Comparison of cone-beam computed tomography and periapical radiography for detecting simulated apical root resorption.

OBJECTIVE: To compare the diagnostic accuracy between cone-beam computed tomography (CBCT) and periapical radiography for detecting simulated external apical root resorption (EARR) in vitro. MATERIALS AND METHODS: The study sample consisted of 160 single-rooted premolar teeth for simulating EARR of varying degrees according to four setups: no (intact teeth), mild (cavity of 1.0 mm in diameter and depth on root surface), moderate (0.4 mm, 0.8 mm, 1.2 mm, and 1.6 mm root shortening), and severe (2.4 mm, 2.8 mm, 3.2 mm, and 3.6 mm root shortening). Two groups of radiographic images were obtained via CBCT and periapical radiography. The absence or presence and the severity for all resorption lesions were evaluated blindly by two calibrated observers. RESULTS: With the CBCT method, the rates of correct classification of no, mild, moderate, and severe EARR were 96.3%, 98.8%, 41.3%, and 87.5%, respectively; with the periapical radiography method, the rates were 82.5%, 41.3%, 68.8%, and 92.5%, respectively. Highly significant differences were found between the two imaging methods for detection of mild (P < .001), moderate (P < .001), and all EARR (P < .001). For detection of all EARR, the sensitivity and specificity values were 75.8% and 96.3% for CBCT, compared with 67.5% and 82.5% for periapical radiography. CONCLUSION: CBCT is a reliable diagnostic tool to detect simulated EARR, whereas periapical radiography underestimates it. However, if a periapical radiograph is already available to the diagnosis of EARR, CBCT should be used with extreme caution to avoid additional radiation exposure.

Simultaneous saccharification and fermentation of fungal pretreated cornstalk for hydrogen production using Thermoanaerobacterium thermosaccharolyticum W16.

In this research, environmentally friendly fungal pretreatment was first adopted for deconstruction of cornstalk. Then the fungal-pretreated cornstalk was employed to produce hydrogen in simultaneous saccharification and fermentation (SSF) using crude enzyme from Trichoderma viride and Thermoanaerobacterium thermosaccharolyticum W16. The influence of various factors including substrate concentration, initial pH, and enzyme loading on hydrogen production were evaluated. The highest hydrogen yield of 89.3 ml/g-cornstalk was obtained with an initial pH 6.5, 0.75% substrate concentration, and 34 FPU/g cellulose. Compared the result with SSF of physical or chemical pretreated lignocellulosic materials, this research suggested an economic and efficient way for hydrogen production from lignocellulosic biomass.