Construction of multifunctional coating with cationic amino acid-coupled peptides for osseointegration of implants.

Osseointegration is an important indicator of implant success. This process can be improved by coating modified bioactive molecules with multiple functions on the surface of implants. Herein, a simple multifunctional coating that could effectively improve osseointegration was prepared through layer-by-layer self-assembly of cationic amino acids and tannic acid (TA), a negatively charged molecule. Osteogenic growth peptide (OGP) and the arginine-glycine-aspartic acid (RGD) functional polypeptides were coupled with Lys6 (K6), the two polypeptides then self-assembled with TA layer by layer to form a composite film, (TA-OGP@RGD)n. The surface morphology and biomechanical properties of the coating were analyzed in gas and liquid phases, and the deposition process and kinetics of the two peptides onto TA were monitored using a quartz crystal microbalance. In addition, the feeding consistency and adsorption ratios of the two peptides were explored by using fluorescence visualization and quantification. The (TA-OGP@RGD)n composite membrane mediated the early migration and adhesion of cells and significantly promoted osteogenic differentiation and mineralization of the extracellular matrix in vitro. Additionally, the bifunctional peptide exhibited excellent osteogenesis and osseointegration owing to the synergistic effect of the OGP and RGD peptides in vivo. Simultaneously, the (TA-OGP@RGD)n membrane regulated the balance of reactive oxygen species in the cell growth environment, thereby influencing the complex biological process of osseointegration. Thus, the results of this study provide a novel perspective for constructing multifunctional coatings for implants and has considerable application potential in orthopedics and dentistry.

NUDCD1 knockdown inhibits the proliferation, migration, and invasion of pancreatic cancer via the EMT process.

NudC domain containing 1 (NUDCD1) is an oncoprotein frequently activated or upregulated in various human cancers, but its role in pancreatic cancer (PC) remains unknown. Thus, we aimed to determine the function and mechanism of NUDCD1 in PC. We employed Western blot and quantitative real-time polymerase chain reaction to assess NUDCD1 expression in cells and PC tissues. NUDCD1 was knocked down in Patu8988 and PANC-1 cells. We conducted real-time cell analysis, wound healing assay, transwell assay and colony formation assay to evaluate the metastatic and proliferative abilities of PC cells. Western blot was conducted to assess the expression of markers associated with apoptosis and epithelial-mesenchymal transition (EMT). Also, we established a tumor xenograft model to determine the role of NUDCD1 in vivo. NUDCD1 was overexpressed in PC tissues and cells. NUDCD1 knockdown suppressed the invasion, migration, and proliferative abilities of the cells and induced PC cell apoptosis. The specific mechanism of NUDCD1 was related to the modulation of the EMT process. Data obtained from in vivo experiments revealed that NUDCD1 knockdown inhibited the tumor growth, proliferation, and metastasis by modulating the EMT and inducing the apoptosis of PC cells.

Relative effectiveness of facemask therapy with alternate maxillary expansion and constriction in the early treatment of Class III malocclusion.

INTRODUCTION: This study aimed to investigate the relative efficacy of maxillary protraction combined with a modified alternate rapid maxillary expansion and constriction (Alt-RAMEC) protocol compared with conventional protocols in the early orthopedic treatment of skeletal Class III malocclusion. METHODS: A sample of 39 patients was divided into 3 groups on the basis of different interventions. Conventional facemask (FM) with splint-type intraoral devices was performed in the FM group (7 males and 5 females; mean age, 9.53 ± 1.37 years). Maxillary expansion with an activation rate of 0.5 mm/d (twice a day) followed by FM therapy was applied in the rapid maxillary expansion group (RME/FM) (6 males and 6 females; mean age, 9.31 ± 1.60 years). In the Alt-RAMEC/FM group (7 males and 8 females; mean age, 10.01 ± 1.31 years), Alt-RAMEC was started simultaneously and throughout the entire course of maxillary protraction, with repetitive alternations between activation and deactivation of expanders (0.5 mm/d for 7 days). The patients in all groups were instructed to wear FMs for a minimum of 12 h/d. Pretreatment and posttreatment lateral cephalograms were all traced and measured. RESULTS: The Alt-RAMEC group showed statistically more significant maxillary advancement than other groups (A-VRP, 3.87 mm vs 3.04 mm [RME/FM], vs 2.04 mm [FM]; P <0.05). Analysis of variance did not reveal significant intergroup differences in palatal plane angulation changes (P >0.05). No pronounced mandibular clockwise rotations were noted in the Alt-RAMEC/FM group with distinct intergroup differences (P <0.05). There were more skeletal effects (88.7%) during overjet correction in the Alt-RAMEC/FM protocol. CONCLUSIONS: A combination of the modified Alt-RAMEC protocol with FM revealed more favorable skeletal effects compared with FM and RME/FM protocols in treating prepubertal patients with maxillary deficiency.

A Bayesian network meta-analysis of orthopaedic treatment in Class III malocclusion: Maxillary protraction with skeletal anchorage or a rapid maxillary expander.

To evaluate and compare the effectiveness of orthopaedic treatment for Class III malocclusions using skeletal anchorage or a rapid maxillary expander for maxillary protraction. Electronic databases, including PubMed, EMBASE, Cochrane Library and Web of Science, were searched for randomized controlled trials (RCTs) and non-randomized clinical trials (CCTs) for orthopaedic treatment of Class III malocclusions. Five interventions were studied: a facemask with a maxillary temporary anchorage device (MTAD), a bone-anchored rapid maxillary expansion (BARME), a rapid maxillary expansion (RME), an alternate rapid maxillary expansion and contraction (Alt-RAMEC), and a bone-anchored intermaxillary traction (BAIMT). Eight outcomes (SNA, SNB, ANB, overjet, SN-GoGn, ANS-Me, IMPA (L1-MP), and U1-PP) were statistically polled. We conducted network meta-analysis using R statistical software with the GeMTC package. Twenty-five studies met the inclusion criteria. Compared with the RME group, the Alt-RAMEC group (mean difference (MD): 1.3; 95% credibility interval (CrI): 0.26, 2.3) and MTAD group (MD: 0.85; 95% CrI: 0.065, 1.6) showed a better effect on ANB in CCTs. Regarding the vertical relationship, the BAIMT group (MD: -2.2; 95% CrI: -5.2, 0.73) showed a smaller effect regarding increasing the vertical dimension of ANS-Me. The RME, MTAD and Alt-RAMEC group showed a higher ability to decrease the angle of L1-MP. The Alt-RAMEC and MTAD protocol have a higher possibility to obtain a skeletal and tooth effect in sagittal relationships. The BAIMT protocol can acquire a better skeletal effect in sagittal relationships with less vertical and dental changes. More well-designed RCTs are needed to ensure that the conclusion is reliable.

Nicotine may promote tongue squamous cell carcinoma progression by activating the Wnt/ß-catenin and Wnt/PCP signaling pathways.

To investigate the effects and the possible underlying mechanisms of nicotine stimulation on tongue squamous cell carcinoma (TSCC) progression, a TSCC cell line Cal27 and 34 samples of paraffin-embedded TSCC were examined. Immunofluorescence, western blot analysis, and TOP/FOP flash, CCK-8, wound healing and Transwell invasion assays were used to evaluate Cal27 in response to nicotine stimulation. We also investigated expression levels of related proteins of Wnt/ß-catenin and Wnt/PCP pathways in paraffin-embedded TSCC samples with or without a history of smoking by immunohistochemistry. Nicotine stimulation can promote proliferation, migration, and invasion of TSCC cells in vitro, downregulate E-cadherin, and activate the Wnt/ß-catenin and Wnt/PCP pathways, which could be antagonized by the α7 nicotine acetylcholine receptor (α7 nAChR) inhibitor α-BTX. Moreover, the expression levels of ß-catenin, Wnt5a and Ror2 were higher in TSCC patients with a history of smoking than those without a history of smoking. Our results suggest nicotine may promote tongue squamous carcinoma cells progression by activating the Wnt/ß-catenin and Wnt/PCP signaling pathways and may play a significant role in the progression and metastasis of smoking-related TSCC.

Forms of nitrogen uptake, translocation, and transfer via arbuscular mycorrhizal fungi: a review.

Arbuscular mycorrhizal (AM) fungi are obligate symbionts that colonize the roots of more than 80% of land plants. Experiments on the relationship between the host plant and AM in soil or in sterile root-organ culture have provided clear evidence that the extraradical mycelia of AM fungi uptake various forms of nitrogen (N) and transport the assimilated N to the roots of the host plant. However, the uptake mechanisms of various forms of N and its translocation and transfer from the fungus to the host are virtually unknown. Therefore, there is a dearth of integrated models describing the movement of N through the AM fungal hyphae. Recent studies examined Ri T-DNA-transformed carrot roots colonized with AM fungi in (15)N tracer experiments. In these experiments, the activities of key enzymes were determined, and expressions of genes related to N assimilation and translocation pathways were quantified. This review summarizes and discusses the results of recent research on the forms of N uptake, transport, degradation, and transfer to the roots of the host plant and the underlying mechanisms, as well as research on the forms of N and carbon used by germinating spores and their effects on amino acid metabolism. Finally, a pathway model summarizing the entire mechanism of N metabolism in AM fungi is outlined.

Germinating spores of Glomus intraradices can use internal and exogenous nitrogen sources for de novo biosynthesis of amino acids.

* Here, nitrogen (N) uptake and metabolism, and related gene expression, were analyzed in germinating spores of Glomus intraradices to examine the mechanisms and the regulation of N handling during presymbiotic growth. * The uptake and incorporation of organic and inorganic N sources into free amino acids were analyzed using stable and radioactive isotope labeling followed by high-performance liquid chromatography (HPLC), gas chromatography-mass spectrometry (GC-MS) and liquid scintillation counting and the fungal gene expression was measured by quantitative polymerase chain reaction (Q-PCR). * Quiescent spores store Asp, Ala and Arg and can use these internal N resources during germination. Although not required for presymbiotic growth, exogenous N can also be utilized for the de novo biosynthesis of amino acids. Ammonium and urea are more rapidly assimilated than nitrate and amino acids. Root exudates do not stimulate the uptake and utilization of exogenous ammonium, but the expression of genes encoding a putative glutamate dehydrogenase (GDH), a urease accessory protein (UAP) and an ornithine aminotransferase (OAT) were stimulated by root exudates. The transcript levels of an ammonium transporter (AMT) and a glutamine synthetase (GS) were not affected. * Germinating spores can make effective use of different N sources and the ability to synthesize amino acids does not limit presymbiotic growth of arbuscular mycorrhizal (AM) spores.

Arginine bi-directional translocation and breakdown into ornithine along the arbuscular mycorrhizal mycelium.

Bi-directional translocation and degradation of Arginine (Arg) along the arbuscular mycorrhizal (AM) fungal mycelium were testified through (15)N and/or (13)C isotopic labeling. In vitro mycorrhizas of Glomus intraradices and Ri T-DNA-transformed carrot roots were grown in dual compartment Petri dishes. [(15)N- and/or(13)C]Arg was supplied to either the fungal compartment or the mycorrhizal compartment or separate dishes containing the uncolonized roots. The levels and labeling of free amino acids (AAs) in the mycorrhizal roots and in the extraradical mycelia(ERM) were measured by gas chromatography/mass spectrometry (GC-MS) and high-performance liquid chromatography (HPLC). The ERM of AM fungi exposed in either NH(4) (+) or urea as sole external nitrogen source had much higher (15)N enrichment of Arg, compared with those in nitrate or exogenous Arg; however, glycerol supplied as an external carbon source to the ERM had no significant effect on the level of Arg in the ERM. Meanwhile, Arg biosynthesized in the ERM could be translocated intact to the mycorrhizal roots and thereby the level of Arg in the mycorrhizal roots increased to about 20% after culture of ERM in 4 mmol/L NH(4) (+) for 6 weeks. Also Arg was found to be bi-directionally transported along the AM fungal mycelium through [U-(13)C]Arg labeling either in the mycorrhizal compartment or in the fungal compartment. Once Arg was translocated to the potential N-limited sites, it would be further degraded into ornithine (Orn) and urea since either [U-(13)C] or [U-(15)N/U-(13)C]Orn was apparently shown up in the mycorrhizal root tissues when [U-(13)C] or [U-(15)N/U-(13)C]Arg was labeled in the fungal compartment, respectively. Evidently Orn formation indicated the ongoing activities of Arg translocation and degradation through the urea cycle in AM fungal mycelium.

Nitrogen transfer in the arbuscular mycorrhizal symbiosis.

Most land plants are symbiotic with arbuscular mycorrhizal fungi (AMF), which take up mineral nutrients from the soil and exchange them with plants for photosynthetically fixed carbon. This exchange is a significant factor in global nutrient cycles as well as in the ecology, evolution and physiology of plants. Despite its importance as a nutrient, very little is known about how AMF take up nitrogen and transfer it to their host plants. Here we report the results of stable isotope labelling experiments showing that inorganic nitrogen taken up by the fungus outside the roots is incorporated into amino acids, translocated from the extraradical to the intraradical mycelium as arginine, but transferred to the plant without carbon. Consistent with this mechanism, the genes of primary nitrogen assimilation are preferentially expressed in the extraradical tissues, whereas genes associated with arginine breakdown are more highly expressed in the intraradical mycelium. Strong changes in the expression of these genes in response to nitrogen availability and form also support the operation of this novel metabolic pathway in the arbuscular mycorrhizal symbiosis.

By-product formation by a novel glycerol-producing yeast, Candida glycerinogenes, with different O2 supplies.

Candida glycerinogenes is an aerobe which does not depend on sulphite for production of glycerol. With a sufficient O2 supply, up to 130 g glycerol l(-1) was produced with 2.6 g acetic acid l(-1) as by-product. However, with an insufficient O2 supply--with higher volumes of medium or at higher corn steep liquid concentrations--the glycerol concentration was lower because the by-products, ethanol, pyruvate and lactic acid, were produced in greater amounts, up to 45 g l(-1), 4.3 g l(-1), 1.6 g l(-1), respectively, whereas, less acetic acid (0.6 g l(-1)) was produced. In addition, ethanol decreased to 0.4 g l(-1) and the glycerol yield improved from 34 to 50% (w/w) by adding 50 g sulphite l(-1), nevertheless, acetic acid increased to 7.8 g l(-1).