Fucoxanthin Inhibits the Proliferation and Metastasis of Human Pharyngeal Squamous Cell Carcinoma by Regulating the PI3K/Akt/mTOR Signaling Pathway.

Human pharyngeal squamous cell carcinoma (HPSCC) is the most common malignancy in the head and neck region, characterized by high mortality and a propensity for metastasis. Fucoxanthin, a carotenoid isolated from brown algae, exhibits pharmacological properties associated with the suppression of tumor proliferation and metastasis. Nevertheless, its potential to inhibit HPSCC proliferation and metastasis has not been fully elucidated. This study represents the first exploration of the inhibitory effects of fucoxanthin on two human pharyngeal squamous carcinoma cell lines (FaDu and Detroit 562), as well as the mechanisms underlying those effects. The results showed dose-dependent decreases in the proliferation, migration, and invasion of HPSCC cells after fucoxanthin treatment. Further studies indicated that fucoxanthin caused a significant reduction in the expression levels of proteins in the phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT)/mechanistic target of rapamycin (mTOR) pathway, as well as the downstream proteins matrix metalloproteinase (MMP)-2 and MMP-9. Specific activators of PI3K/AKT reversed the effects of fucoxanthin on these proteins, as well as on cell proliferation and metastasis, in FaDu and Detroit 562 cells. Molecular docking assays confirmed that fucoxanthin strongly interacted with PI3K, AKT, mTOR, MMP-2, and MMP-9. Overall, fucoxanthin, a functional food component, is a potential therapeutic agent for HPSCC.

Synthesis of Sialidase-Resistant Oligosaccharide and Antibody Glycoform Containing α2,6-Linked 3Fax-Neu5Ac.

Fluorinated glycosides are known to resist the glycosidase-catalyzed glycosidic bond cleavage; however, the synthesis of such glycans, especially 3-fluoro-sialic acid (3F-Neu5Ac) containing sialosides, has been a major challenge. Though the enzymatic synthesis of α-2,3-linked 3F-sialosides was reported, until recently there has not been any effective method available for the synthesis of 3F-sialosides in the α-2,6-linkage. In order to understand the biological effect of such modification, we report here a chemical synthesis of 3Fax-Neu5Ac-α2,6-Gal as a building block for the assembly of 3Fax-Neu5Ac-containing sialosides and a representative homogeneous antibody glycoform. Our results showed that the sialosides are stable under sialidase catalysis and the rituximab glycoform with a sialylated complex-type biantennary glycan terminated with 3Fax-Neu5Ac in the α-2,6-linkage (α2,6-F-SCT) has a similar binding avidity as its parent glycoform. These findings open up new opportunities for the development of therapeutic glycoproteins with improved pharmacokinetic parameters.

Dietary administration of Bacillus subtilis HAINUP40 enhances growth, digestive enzyme activities, innate immune responses and disease resistance of tilapia, Oreochromis niloticus.

The probiotic properties of Bacillus subtilis HAINUP40 isolated from the aquatic environment, and the effects of dietary administration of B. subtilis HAINUP40 on the growth performance, intestinal probiotic recovery, digestive enzyme activities, innate immunity and disease resistance of tilapia (Oreochromis niloticus) were evaluated. The probiotic properties investigated include tolerance to simulated gastrointestinal stress, auto-aggregation, cell surface hydrophobicity and extracellular enzyme production. The cell number of B. subtilis changed little after 4 h in simulated gastric fluid at pH = 2.0, 3.0, 4.0 and simulated intestinal fluid at pH = 6.8.B.subtilis HAINUP40 revealed strong auto-aggregation property (34.6-87.0%) after 24 h incubation period. It exhibited significant cell surface hydrophobicity in xylene (28.8%) and chloroform (41.3%) and produced extracellular proteases and amylase. After tilapia (mean weight = 95 ± 8 g) were fed with a diet containing 108 cfu/g B. subtilis HAINUP40, their final body weight, percent weight gain (PWG), specific growth rate (SGR), total antioxidant capacity (T-AOC) and serum superoxide dismutase (SOD) increased significantly (p < 0.05) after 8 weeks; feed conversion rate (FCR) is significantly lower (p < 0.05) after 8 weeks; the protease and amylase activity in the digestive tract increased significantly (p < 0.05) after 4 and 8 weeks; and respiratory bursts and serum lysozyme activity increased significantly (p < 0.05) after 2 weeks. Moreover, being challenged with pathogenic Streptococcus agalactiae for 2 weeks, the relative percent survival (RPS%) is 52.94%. The results of this study strongly suggest that dietary supplement of B. subtilis HAINUP40 can effectively enhances the growth performance, immune response, and disease resistance of Nile tilapia.

Molecular mechanism of the sweet taste enhancers.

Positive allosteric modulators of the human sweet taste receptor have been developed as a new way of reducing dietary sugar intake. Besides their potential health benefit, the sweet taste enhancers are also valuable tool molecules to study the general mechanism of positive allosteric modulations of T1R taste receptors. Using chimeric receptors, mutagenesis, and molecular modeling, we reveal how these sweet enhancers work at the molecular level. Our data argue that the sweet enhancers follow a similar mechanism as the natural umami taste enhancer molecules. Whereas the sweeteners bind to the hinge region and induce the closure of the Venus flytrap domain of T1R2, the enhancers bind close to the opening and further stabilize the closed and active conformation of the receptor.

Clinical application of modified hip joint lateral position in femoral neck fracture.

BACKGROUND: To show the femoral neck better in hip lateral view of X-ray, we design a modified hip lateral view and then investigate the value in femoral neck fractures. METHODS: CT images of 10 normal hip joints for 3D reconstruction were selected, the Mimics Medical 21.0 was used, and rotating the proximal femur was to find the most suitable angle for showing the femoral neck well, designed the modified lateral view according to this angle. We collected 35 healthy cases and 35 femoral neck fractures as the normal and fracture group. And two groups were all taken hip anteroposterior view, cross-table lateral view and modified lateral view, which were analyzed by two radiologists to score the anatomical structures of the articular surface, femoral head, head neck junction, femoral neck, basal region and intertrochanteric region. Friedman test was used to analyze the score of femoral neck at different angles. T test and Wilcoxon signed-rank test were to compare inter-groups. RESULTS: The modified lateral view was designed as follows: The subjects were supine, with the sagittal axis biased toward the healthy side at an angle of approximately 20° to the long axis of the examination table, the hip joint flexed at 45°, the lower extremity abducted at 40°, the centerline inclined 45° toward the head and the centerline aligned with the center of the groin. The modified lateral view showed the femoral head, head neck junction and femoral neck more clearly than the cross-table lateral view, but the cross-table lateral view showed the femoral neck basal and intertrochanteric region better. In addition, the time of taking the modified lateral view was significantly less than the cross-table lateral view (normal group: 0.789 min ± 0.223 vs 0.623 min ± 0.207, P < 0.001; fracture group: 1.131 min ± 0.362 vs 0.946 min ± 0.390, P < 0.001). CONCLUSIONS: The modified lateral view can obtain a standard sagittal image of femoral neck, which can show the dislocation and angulation of the sagittal femoral neck fracture clearly, and improve the accuracy of diagnosis. And it is more convenient and easier for patients to cooperate, which is worthy promoting and applying in clinical work.

Multiparametric MRI combined with liver volume for quantitative evaluation of liver function in patients with cirrhosis.

PURPOSE We aimed to establish a liver function evaluation model by combining multiparametric magnetic resonance imaging (MRI) with liver volume (LV) and further verify the effectiveness of the model to evaluate liver function. METHODS This retrospective study included 101 consecutive cirrhosis patients (69 cases for modeling group and 32 cases for validation group) who underwent gadoxetic acid-enhanced MRI. Five signal intensity parameters were obtained by measuring the signal intensities of the liver, spleen, and erector spinae before and 20 minutes after gadoxetic acid disodium enhancement. The dif fusion coefficient (D), pseudo-diffusion coefficient (D*), and perfusion fraction (f) were obtained from intravoxel incoherent motion diffusion-weighted imaging. The LV parameters (Vliver, Vspleen, and Vliver/Vspleen) were obtained using 3-dimensional image generation software. The most effec tive parameter was selected from each of the 3 methods, and a multivariate regression model for liver function evaluation was established and validated. RESULTS In the modeling group, relative enhancement (RE), D*, and Vliver/Vspleen showed significant dif ferences among the different liver function groups (P < .001). Receiver operating characteristic analysis showed that these parameters had the highest area under the curve (AUC) values for dis tinguishing Child-Pugh A from Child-Pugh B and C groups (0.917, 0.929, and 0.885, respectively). The following liver function model was obtained by multivariate regression analysis: F(x)=3.96 - 1.243 (RE) - 0.034 (D*) - 0.080 (Vliver/Vspleen) (R2=0.811, P < .001). In the patients with cirrhosis, the F(x) of Child-Pugh A, B, and C were 1.16 ± 0.44, 1.95 ± 0.29, and 2.79 ± 0.38, respectively. In the validation group, the AUC for F(x) to distinguish Child-Pugh A from Child-Pugh B and C was 0.973. CONCLUSION Combining multiparametric MRI with LV effectively distinguished patients with different Child Pugh grades. This model could hence be useful as a novel radiological marker to estimate the liver function.

Is the NH4 +-induced growth inhibition caused by the NH4 + form of the nitrogen source or by soil acidification?

Soil acidification often occurs when the concentration of ammonium (NH4 +) in soil rises, such as that observed in farmland. Both soil acidification and excess NH4 + have serious adverse effects on crop growth and food production. However, we still do not know which of these two inhibitors has a greater impact on the growth of crops, and the degree of their inhibitory effect on crop growth have not been accurately evaluated. 31 wheat cultivars originating in various areas of China were planted under 5 mM sole NH4 + (ammonium nitrogen, AN) or nitrate nitrogen in combined with two pH levels resembling acidified conditions (5.0 and 6.5). The results showed that the shoots and roots biomass were severely reduced by AN in both and these reduction effects were strengthened by a low medium pH. The concentration of free NH4 + and amino acids, the glutamine synthetase activity were significantly higher, but the total soluble sugar content was reduced under NH4 + conditions, and the glutamine synthetase activity was reduced by a low medium pH. Cultivar variance was responsible for the largest proportion of the total variance in plant dry weight, leaf area, nodal root number, total root length and root volume; the nitrogen (N) form explains most of the variation in N and C metabolism; the effects of pH were the greatest for plant height and root average diameter. So, soil acidification and excess NH4 + would cause different degrees of inhibition effects on different plant tissues. The findings are expected to be useful for applying effective strategies for reducing NH4 + stress in the field.

Molecular mechanism for the umami taste synergism.

Umami is one of the 5 basic taste qualities. The umami taste of L-glutamate can be drastically enhanced by 5' ribonucleotides and the synergy is a hallmark of this taste quality. The umami taste receptor is a heteromeric complex of 2 class C G-protein-coupled receptors, T1R1 and T1R3. Here we elucidate the molecular mechanism of the synergy using chimeric T1R receptors, site-directed mutagenesis, and molecular modeling. We propose a cooperative ligand-binding model involving the Venus flytrap domain of T1R1, where L-glutamate binds close to the hinge region, and 5' ribonucleotides bind to an adjacent site close to the opening of the flytrap to further stabilize the closed conformation. This unique mechanism may apply to other class C G-protein-coupled receptors.

Application of BOLD-MRI in the classification of renal function in chronic kidney disease.

PURPOSE: The purpose of the study was to explore the application of blood oxygenation level-dependent functional magnetic resonance imaging (BOLD-MRI) in classification of chronic kidney disease (CKD). METHODS: Twenty-nine cases with CKD and 27 healthy volunteers underwent renal BOLD-MRI. Cases of CKD were divided into two groups according to the estimated glomerular filtration rate (eGFR). The R2* values were measured in renal cortex and medulla, respectively. The difference of R2* between renal cortex and medulla was compared, and the correlations of R2* value in renal cortex and medulla with eGFR were analyzed. RESULTS: Twenty-nine cases of CKD were divided into two groups, with 13 cases of mild renal impairment and 16 cases of moderate to severe renal impairment. In the control and mild renal impairment group, the R2* of renal cortex was significantly lower than that of medulla (P < 0.001). In the control group, mild renal impairment and moderate to severe renal impairment group, the R2* value of cortex increased, while the R2* value of medulla gradually decreased. The eGFR of patients was positively correlated with R2* of medulla (r = 0.81, P < 0.001), while displayed no correlation with R2* of cortex (r = - 0.32, P > 0.05). When the threshold of R2* of medulla was set at 28.4 Hz, the sensitivity and specificity to distinguish normal and mild renal impairment group were 92.31% and 85.19%, respectively. CONCLUSION: The change of blood oxygen in renal cortex and medulla could be detected with BOLD-MRI, so as to evaluate the renal function and anoxic injury of CKD.

Diffusion-weighted imaging for staging chronic kidney disease: a meta-analysis.

OBJECTIVE:: To evaluate stages of chronic kidney disease (CKD) by apparent diffusion coefficient (ADC) obtained from diffusion weighted imaging (DWI) using a meta-analysis. METHODS:: Literature databases were searched from PubMed, Web of Science, Cochrane and Embase to identify relevant articles about DWI in CKD between 1999 and 2017. ADC values were extracted from the healthy group and CKD patients with different stages. Meta-analysis was conducted using STATA v. 12.0. A random-effects model was performed to acquire the effect estimate, which was expressed as a pooled weighted mean difference (WMD) with 95% confidence interval (CI). We performed comparisons of ADC values between the following groups: (1) the ADC values of the normal kidneys vs earlier Stage 1-2 of CKD; (2) Stage 3 vs the Stage 1-2 of CKD; (3) the Stage 4-5 vs the Stage 3. RESULTS:: Six studies were included in this meta-analysis. The CKD patients with earlier Stage 1-2 showed lower ADC values than the healthy subjects [WMD = -0.09, 95% CI(-0.12 to -0.06), p < 0.001]. However, no obvious difference in ADC values was found between the Stage 3 and Stage1-2 of CKD [WMD = -0.09, 95% CI (-0.18 to 0.01), p = 0.08]. The CKD Stage3 had higher ADC values than those of Stage4-5 [WMD = -0.21, 95% CI (-0.32 to -0.11), p = 0.01]. CONCLUSION:: DWI is an accurate and non-invasive imaging technique for early diagnosis and staging of CKD. Quantitative DWI may potentially play a role in making clinical decisions in the follow-up of CKD. ADVANCES IN KNOWLEDGE: DWI can be a valuable tool for staging of CKD.

Characterization of a transglycosylase domain of Streptococcus pneumoniae PBP1b.

Inhibitors of transglycosylases may serve as potent antibiotics that are less prone to resistance development in bacterial pathogens. To facilitate the search of such compounds, a transglycosylase (TGase) domain of the membrane integral multidomain Streptococcus pneumoniae PBP1b was cloned and expressed. This TGase domain was characterized by a substrate-dependent fluorescence coupled enzyme assay and an inhibitor-tethered surface plasmon resonance binding assay. Both assays show that the catalytic efficiency of the domain is comparable to that of the monofunctional transglycosylases, and it is fully active in the absence of other domains. The isolation of the active TGase domain makes it possible to screen for potential antibiotics targeting transglycosylases.

A method for the generation of glycoprotein mimetics.

A general method for preparing glycoprotein mimetics with defined glycan structure using the Z domain protein as an example is reported. An unnatural amino acid containing the keto group was site-specifically incorporated into a target protein, Z domain, in response to the amber nonsense codon with high translational fidelity and efficiency. An aminooxy saccharide derivative was then selectively coupled to this genetically encoded keto group. The resulting saccharide core was elaborated to a glycan complex with glycosyltransferases. Alternatively, aminooxy analogues of more complicated glycans were prepared and directly attached to the keto group. Homogeneous glycoprotein mimetics thus prepared should prove useful for the study of carbohydrate effects on glycoprotein structure and function. This method may also lead to the production of glycoprotein therapeutics from Escherichia coli.

Acceptor specificity and inhibition of the bacterial cell-wall glycosyltransferase MurG.

A continuous fluorescence coupled enzyme assay was developed to study the acceptor specificity of the glycosyltransferase MurG toward different lipid I analogues with various substituents replacing the undecaprenyl moiety. It was found that most lipid I analogues are accepted as substrates and, amongst these, the saturated C14 analogue exhibits the best activity. This substrate was used to evaluate the inhibition activity of such antibiotics as moenomycin, vancomycin, and two chlorobiphenyl vancomycin derivatives. A vancomycin derivative with a chlorobiphenyl moiety on the aglycon section was identified as a potent inhibitor of MurG.

Synthesis and high-throughput screening of N-acetyl-beta-hexosaminidase inhibitor libraries targeting osteoarthritis.

C1 Nitrogen iminocyclitols are potent inhibitors of N-acetyl-beta-hexosaminidases. Given hexosaminidases' important roles in osteoarthritis, we developed two straightforward and efficient syntheses of C1 nitrogen iminocyclitols from two readily available starting materials, D-mannosamine hydrochloride and the microbial oxidation product of fructose. A diversity-oriented synthetic strategy was then performed by coupling these core structures with various aldehydes, carboxylic acids, and alkynes to generate three separate libraries. High-throughput screening of the generated libraries with human N-acetyl-beta-hexosaminidases produced only moderate inhibitory activities. However, the synthetic approach and screening strategy for these compounds will be applied to develop new potent inhibitors of human N-acetyl-beta-hexosaminidases, particularly when combined with the structural information of these enzymes.

Cell-wall engineering of living bacteria.

The cell walls of living bacteria were chemically modified by adding cell-wall precursors. As the precursors to be incorporated into the cell wall, UDP-MurNAc pentapeptide, lipid I, and lipid II derivatives were synthesized. The aimed compounds were attached to the amine residue of lysine at the pentapeptide moiety. Fluorescein-attached UDP-MurNAc pentapeptide was efficiently incorporated into both Gram-positive and Gram-negative bacteria. In the case of Gram-negative bacteria, such as Escherichia coli, the permeability of the outer membrane (lipopolysaccharide layer) was enhanced by EDTA treatment before the incorporation. For Gram-positive bacteria, UDP-MurNAc derivatives were incorporated in the cell wall without EDTA treatment due to the lack of the lipopolysaccharide layer. Furthermore, instead of dyes, a ketone group was attached to the UDP-MurNAc pentapeptide. The ketone group was also delivered to the bacterial cell wall of lactic acid bacteria, giving a platform to attach large molecules on the surface.