Multivalent dipeptidyl peptidase IV fragment-nanogold complex inhibits cancer metastasis by blocking pericellular fibronectin.

Inhibition of cancer metastasis is a fundamental challenge in cancer treatment. We have previously shown that metastasis of cancer cells in the lung is critically promoted by the interaction between the superficial dipeptidyl peptidase IV (DPP IV) expressed on lung endothelial cells and the pericellular polymeric fibronectin (polyFN) of circulating cancer cells. In the present study, we aimed to search for DPP IV fragments with high avidity to polyFN and develop FN-targeted gold nanoparticles (AuNPs) conjugated with DPP IV fragments for treating cancer metastasis. We first identified a DPP IV fragment encompassing amino acids 29-130 of DPP IV, designated DP4A, which contained FN-binding sites and could specifically bind to FN immobilized on gelatin agarose beads. Furthermore, we conjugated maltose binding protein (MBP)-fused DP4A proteins to AuNPs for fabricating a DP4A-AuNP complex and evaluated its FN-targeted activity in vitro and anti-metastatic efficacy in vivo. Our results show that DP4A-AuNP exhibited higher binding avidity to polyFN than DP4A by 9 folds. Furthermore, DP4A-AuNP was more potent than DP4A in inhibiting DPP IV binding to polyFN. In terms of polyFN-targeted effect, DP4A-AuNP interacted with FN-overexpressing cancer cells and was endocytosed into cells 10 to 100 times more efficiently than untargeted MBP-AuNP or PEG-AuNP with no noticeable cytotoxicity. Furthermore, DP4A-AuNP was superior to DP4A in competitive inhibition of cancer cell adhesion to DPP IV. Confocal microscopy analysis revealed that binding of DP4A-AuNP to pericellular FN induced FN clustering without altering its surface expression on cancer cells. Notably, intravenous treatment with DP4A-AuNP significantly reduced metastatic lung tumor nodules and prolonged the survival in the experimental metastatic 4T1 tumor model. Collectively, our findings suggest that the DP4A-AuNP complex with potent FN-targeted effects may have therapeutic potential for prevention and treatment of tumor metastasis to the lung.

Genotypic Diversity of Ciprofloxacin Nonsusceptibility and Its Relationship with Minimum Inhibitory Concentrations in Nontyphoidal Salmonella Clinical Isolates in Taiwan.

This study analyzed the genetic diversity of ciprofloxacin (CIP) nonsusceptibility and the relationship between two major mechanisms and minimum inhibitory concentrations (MICs) of CIP in nontyphoidal Salmonella (NTS). Chromosomal mutations in quinolone resistance-determining regions (QRDRs) and plasmid-mediated quinolone resistance (PMQR) genes were searched from ResFinder, ARG-ANNOT, and PubMed for designing the sequencing regions in gyrA, gyrB, parC, and parE, and the 13 polymerase chain reactions for PMQR genes. We found that QRDR mutations were detected in gyrA (82.1%), parC (59.0%), and parE (20.5%) but not in gyrB among the 39 isolates. Five of the 13 PMQR genes were identified, including oqxA (28.2%), oqxB (28.2%), qnrS (18.0%), aac(6')-Ib-cr (10.3%), and qnrB (5.1%), which correlated with the MICs of CIP within 0.25-2 µg/mL, and it was found that oxqAB contributed more than qnr genes to increase the MICs. All the isolates contained either QRDR mutations (53.8%), PMQR genes (15.4%), or both (30.8%). QRDR mutations (84.6%) were more commonly detected than PMQR genes (46.2%). QRDR mutation numbers were significantly associated with MICs (p < 0.001). Double mutations in gyrA and parC determined high CIP resistance (MICs ≥ 4 µg/mL). PMQR genes contributed to intermediate to low CIP resistance (MICs 0.25-2 µg/mL), thus providing insights into mechanisms underlying CIP resistance.

Utility of quantitative ultrasound in community screening for hepatic steatosis.

Nonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disease. Quantitative ultrasound facilitates clinical grading of hepatic steatosis (the early stage of NAFLD). However, the utility of quantitative ultrasound as a first-line method for community screening of hepatic steatosis remains unclear. Therefore, this study aimed to investigate the utility of quantitative ultrasound to screen for hepatic steatosis and for metabolic evaluation at the community level. In total, 278 participants enrolled from a community satisfied the study criteria. Each subject underwent anthropometric and biochemical examinations, and abdominal ultrasound imaging was performed to measure the controlled attenuation (CAP), integrated backscatter (IB), and information Shannon entropy (ISE). The assessment outcomes were compared with the fatty liver index (FLI), hepatic steatosis index (HSI), metabolic syndrome (MetS), and insulin resistance to evaluate the screening performance through the area under the receiver operating characteristic curve (AUROC) and Delong's test. Ultrasound ISE, CAP, and IB were effective in screening hepatic steatosis, MetS, and insulin resistance. In screening for hepatic steatosis, the AUROCs of ISE, CAP, and IB were 0.85, 0.83, and 0.80 (the cutoff FLI = 60), respectively, and 0.84, 0.75, 0.77 (the cutoff HSI = 36), respectively, and those for the evaluation of MetS and insulin resistance were 0.79, 0.75, 0.79, respectively, and 0.83, 0.76, 0.78, respectively. Delong's test revealed that ISE outperformed CAP and IB for the detection of hepatic steatosis and insulin resistance (P < .05). Based on the present results, ultrasound ISE is a potential imaging biomarker during first-line community screening of hepatic steatosis and insulin resistance.

Characterization of limb lymphedema using the statistical analysis of ultrasound backscattering.

BACKGROUND: Lymphedema is a disease in which tissue swelling is caused by interstitial fluid retention in subcutaneous tissue. It is caused by a compromised lymphatic system. Lymphoscintigraphy is the current and primary modality used to assess lymphatic system dysfunction. Ultrasound elastography is a complementary tool used for evaluating the tissue stiffness of the lymphedematous limb. Tissue stiffness implies the existence of changes in tissue microstructures. However, ultrasound features related to tissue microstructures are neglected in clinical assessments of lymphedematous limbs. In this study, we aimed to evaluate the lymphedematous diagnostic values of ultrasound Nakagami and entropy imaging, which are, respectively, model- and nonmodel-based backscattered statistical analysis methods for scatterer characterization. METHODS: A total of 60 patients were recruited, and lymphoscintigraphy was used to score the patient's clinical severity of each of their limb lymphedema (0: normal; 1: partial lymphatic obstruction; and 2: total lymphatic obstruction). We performed ultrasound examinations to acquire ultrasound backscattered signals for B-mode, Nakagami, and entropy imaging. The envelope amplitude, Nakagami, and entropy values, as a function of the patients' lymphatic obstruction grades, were expressed in terms of their median and interquartile range (IQR). The values were then used in both an independent t test and a receiver operating characteristic (ROC) curve analysis. RESULTS: For each increase in a patient's score from 0 to 2, the envelope amplitude values were 405.44 (IQR: 238.72-488.17), 411.52 (IQR: 298.53-644.25), and 476.37 (IQR: 348.86-648.16), respectively. The Nakagami parameters were 0.16 (IQR: 0.14-0.22), 0.26 (IQR: 0.23-0.34), and 0.24 (IQR: 0.16-0.36), respectively, and the entropy values were 4.55 (IQR: 4.41-4.66), 4.86 (IQR: 4.78-4.99), and 4.87 (IQR: 4.81-4.97), respectively. The P values between the normal control and lymphedema groups obtained from B-mode and Nakagami analysis were larger than 0.05; whereas that of entropy imaging was smaller than 0.05. The areas under the ROC curve for B-mode, Nakagami, and entropy imaging were 0.64 (sensitivity: 70%; specificity: 47.5%), 0.75 (sensitivity: 70%; specificity: 75%), and 0.94 (sensitivity: 95%; specificity: 87.5%), respectively. CONCLUSIONS: The current findings demonstrated the diagnostic values of ultrasound Nakagami and entropy imaging techniques. In particular, the use of non-model-based entropy imaging enables for improved performance when characterizing limb lymphedema.

Value of homodyned K distribution in ultrasound parametric imaging of hepatic steatosis: An animal study.

Ultrasound is the first-line tool for screening hepatic steatosis. Statistical distributions can be used to model the backscattered signals for liver characterization. The Nakagami distribution is the most frequently adopted model; however, the homodyned K (HK) distribution has received attention due to its link to physical meaning and improved parameter estimation through X- and U-statistics (termed "XU"). To assess hepatic steatosis, we proposed HK parametric imaging based on the α parameter (a measure of the number of scatterers per resolution cell) calculated using the XU estimator. Using a commercial system equipped with a 7-MHz linear array transducer, phantom experiments were performed to suggest an appropriate window size for α imaging using the sliding window technique, which was further applied to measuring the livers of rats (n = 66) with hepatic steatosis induced by feeding the rats a methionine- and choline-deficient diet. The relationships between the α parameter, the stage of hepatic steatosis, and histological features were verified by the correlation coefficient r, one-way analysis of variance, and regression analysis. The phantom results showed that the window side length corresponding to five times the pulse length supported a reliable α imaging. The α parameter showed a promising performance for grading hepatic steatosis (p < 0.05; r2 = 0.68). Compared with conventional Nakagami imaging, α parametric imaging provided significant information associated with fat droplet size (p < 0.05; r2 = 0.53), enabling further analysis and evaluation of severe hepatic steatosis.

Considerations of Ultrasound Scanning Approaches in Non-alcoholic Fatty Liver Disease Assessment through Acoustic Structure Quantification.

Non-alcoholic fatty liver disease (NAFLD) is a risk factor for hepatic fibrosis and cirrhosis. Acoustic structure quantification (ASQ), based on statistical analysis of ultrasound echoes, is an emerging technique for hepatic steatosis diagnosis. A standardized measurement protocol for ASQ analysis was suggested previously; however, an optimal ultrasound scanning approach has not been concluded thus far. In this study, the suitability of scanning approaches for the ASQ-based evaluation of hepatic steatosis was investigated. Hepatic fat fractions (HFFs; liver segments VIII, III and VI) of 70 living liver donors were assessed with magnetic resonance spectroscopy. A clinical ultrasound machine equipped with a 3-MHz convex transducer was used to scan each participant using the intercostal, epigastric and subcostal planes to acquire raw data for estimating two ASQ parameters (Cm2 and focal disturbance [FD] ratio) of segments VIII, III and VI, respectively. The parameters were plotted as functions of the HFF for calculating the values of the correlation coefficient (r) and probability value (p). The diagnostic performance of the parameters in discriminating between the normal and steatotic (≥5 and ≥10%) groups was also compared using receiver operating characteristic (ROC) curves. The Cm2 and FD ratio values measured using the epigastric and subcostal planes did not correlate with the severity of hepatic steatosis. However, intercostal imaging exhibited a higher correlation between the ASQ parameters and HFF (r = -0.64, p < 0.001). The diagnostic performance of Cm2 and FD ratio in detecting hepatic steatosis using intercostal imaging was also satisfactory (areas under ROC curves >0.8). Intercostal imaging is an appropriate scanning approach for ASQ analysis of the liver.

Hepatic steatosis assessment using ultrasound homodyned-K parametric imaging: the effects of estimators.

BACKGROUND: The homodyned-K (HK) distribution is an important statistical model for describing ultrasound backscatter envelope statistics. HK parametric imaging has shown potential for characterizing hepatic steatosis. However, the feasibility of HK parametric imaging in assessing human hepatic steatosis in vivo remains unclear. METHODS: In this paper, ultrasound HK µ parametric imaging was proposed for assessing human hepatic steatosis in vivo. Two recent estimators for the HK model, RSK (the level-curve method that uses the signal-to-noise ratio (SNR), skewness, and kurtosis based on the fractional moments of the envelope) and XU (the estimation method based on the first moment of the intensity and two log-moments, namely X- and U-statistics), were investigated. Liver donors (n=72) and patients (n=204) were recruited to evaluate hepatic fat fractions (HFFs) using magnetic resonance spectroscopy and to evaluate the stages of fatty liver disease (normal, mild, moderate, and severe) using liver biopsy with histopathology. Livers were scanned using a 3-MHz ultrasound to construct µ RSK and µ XU images to correlate with HFF analyses and fatty liver stages. The µ RSK and µ XU parametric images were constructed using the sliding window technique with the window side length (WSL) =1-9 pulse lengths (PLs). The diagnostic values of the µ RSK and µ XU parametric imaging methods were evaluated using receiver operating characteristic (ROC) curves. RESULTS: For the 72 participants in Group A, the µ RSK parametric imaging with WSL =2-9 PLs exhibited similar correlation with log10(HFF), and the µ RSK parametric imaging with WSL = 3 PLs had the highest correlation with log10(HFF) (r=0.592); the µ XU parametric imaging with WSL =1-9 PLs exhibited similar correlation with log10(HFF), and the µ XU parametric imaging with WSL =1 PL had the highest correlation with log10(HFF) (r=0.628). For the 204 patients in Group B, the areas under the ROC (AUROCs) obtained using µ RSK for fatty stages ≥ mild (AUROC1), ≥ moderate (AUROC2), and ≥ severe (AUROC3) were (AUROC1, AUROC2, AUROC3) = (0.56, 0.57, 0.53), (0.68, 0.72, 0.75), (0.73, 0.78, 0.80), (0.74, 0.77, 0.79), (0.74, 0.78, 0.79), (0.75, 0.80, 0.82), (0.74, 0.77, 0.83), (0.74, 0.78, 0.84) and (0.73, 0.76, 0.83) for WSL =1, 2, 3, 4, 5, 6, 7, 8 and 9 PLs, respectively. The AUROCs obtained using µ XU for fatty stages ≥ mild, ≥ moderate, and ≥ severe were (AUROC1, AUROC2, AUROC3) = (0.75, 0.83, 0.81), (0.74, 0.80, 0.80), (0.76, 0.82, 0.82), (0.74, 0.80, 0.84), (0.76, 0.80, 0.83), (0.75, 0.80, 0.84), (0.75, 0.79, 0.85), (0.75, 0.80, 0.85) and (0.73, 0.77, 0.83) for WSL = 1, 2, 3, 4, 5, 6, 7, 8 and 9 PLs, respectively. CONCLUSIONS: Both the µ RSK and µ XU parametric images are feasible for evaluating human hepatic steatosis. The WSL exhibits little impact on the diagnosing performance of the µ RSK and µ XU parametric imaging. The µ XU parametric imaging provided improved performance compared to the µ RSK parametric imaging in characterizing human hepatic steatosis in vivo.

Characterization of stm3030 and stm3031 genes of Salmonella enterica serovar Typhimurium in relation to cephalosporin resistance.

BACKGROUND/PURPOSE: The outer membrane protein STM3031 had been shown to confer Salmonella enterica serovar Typhimurium resistance to ceftriaxone. In this study, the STM3030 was increased in strain R200 and decreased in strain R200(Δstm3031). How stm3030 and stm3031 contributing to antibiotic resistance was investigated. METHODS: The level of STM3030 protein in R200(Δstm3031) were compared between 01-4, R200, and R200(Δstm3031) by 2-DE analysis. The stm3030 gene deleted strain, R200(Δstm3030), was generated by the one-step inactivation chromosome gene method. The various antibiotic susceptibility of strains 01-4, R200, R200(Δstm3031) and R200(Δstm3030) were determined by agar dilutions assays and E-test. The co-transcription of stm3031 and stm3030 were determined by RT-PCR. The promoter activities of these two genes fused with LacZ were determined. The binding of the regulatory protein BaeR on the promoter of both genes was detected by EMSA. The interaction between STM3030 and STM3031 proteins was determined by GST pull-down assay. RESULTS: Strain R200(Δstm3030) displayed a 32- to 64-fold reduction in resistance to cephalosporin drugs. Transcription analyses revealed that stm3030 and stm3031 are independent genes and that the promoter of stm3030 is stronger than that of stm3031. The regulator BaeR binds to the promoter region of stm3031 but not that of stm3030. The STM3031 decreased in R200(Δstm3030) compared to R200 by western blot analysis. The pull-down assay revealed that STM3030 and STM3031 bind to each other. CONCLUSION: Our data indicate that STM3030 has a chaperone-like activity and may modulate or stabilize STM3031, leading to resistance of S. enterica serovar Typhimurium to cephalosporin drugs.

Automated tongue diagnosis on the smartphone and its applications.

Tongue features are important objective basis for clinical diagnosis and treatment in both western medicine and Chinese medicine. The need for continuous monitoring of health conditions inspires us to develop an automatic tongue diagnosis system based on built-in sensors of smartphones. However, tongue images taken by smartphone are quite different in color due to various lighting conditions, and it consequently affects the diagnosis especially when we use the appearance of tongue fur to infer health conditions. In this paper, we captured paired tongue images with and without flash, and the color difference between the paired images is used to estimate the lighting condition based on the Support Vector Machine (SVM). The color correction matrices for three kinds of common lights (i.e., fluorescent, halogen and incandescent) are pre-trained by using a ColorChecker-based method, and the corresponding pre-trained matrix for the estimated lighting is then applied to eliminate the effect of color distortion. We further use tongue fur detection as an example to discuss the effect of different model parameters and ColorCheckers for training the tongue color correction matrix under different lighting conditions. Finally, in order to demonstrate the potential use of our proposed system, we recruited 246 patients over a period of 2.5 years from a local hospital in Taiwan and examined the correlations between the captured tongue features and alanine aminotransferase (ALT)/aspartate aminotransferase (AST), which are important bio-markers for liver diseases. We found that some tongue features have strong correlation with AST or ALT, which suggests the possible use of these tongue features captured on a smartphone to provide an early warning of liver diseases.

Ultrasound Entropy Imaging of Nonalcoholic Fatty Liver Disease: Association with Metabolic Syndrome.

Nonalcoholic fatty liver disease (NAFLD) is the leading cause of advanced liver diseases. Fat accumulation in the liver changes the hepatic microstructure and the corresponding statistics of ultrasound backscattered signals. Acoustic structure quantification (ASQ) is a typical model-based method for analyzing backscattered statistics. Shannon entropy, initially proposed in information theory, has been demonstrated as a more flexible solution for imaging and describing backscattered statistics without considering data distribution. NAFLD is a hepatic manifestation of metabolic syndrome (MetS). Therefore, we investigated the association between ultrasound entropy imaging of NAFLD and MetS for comparison with that obtained from ASQ. A total of 394 participants were recruited to undergo physical examinations and blood tests to diagnose MetS. Then, abdominal ultrasound screening of the liver was performed to calculate the ultrasonographic fatty liver indicator (US-FLI) as a measure of NAFLD severity. The ASQ analysis and ultrasound entropy parametric imaging were further constructed using the raw image data to calculate the focal disturbance (FD) ratio and entropy value, respectively. Tertiles were used to split the data of the FD ratio and entropy into three groups for statistical analysis. The correlation coefficient r, probability value p, and odds ratio (OR) were calculated. With an increase in the US-FLI, the entropy value increased (r = 0.713; p < 0.0001) and the FD ratio decreased (r = -0.630; p < 0.0001). In addition, the entropy value and FD ratio correlated with metabolic indices (p < 0.0001). After adjustment for confounding factors, entropy imaging (OR = 7.91, 95% confidence interval (CI): 0.96-65.18 for the second tertile; OR = 20.47, 95% CI: 2.48-168.67 for the third tertile; p = 0.0021) still provided a more significant link to the risk of MetS than did the FD ratio obtained from ASQ (OR = 0.55, 95% CI: 0.27-1.14 for the second tertile; OR = 0.42, 95% CI: 0.15-1.17 for the third tertile; p = 0.13). Thus, ultrasound entropy imaging can provide information on hepatic steatosis. In particular, ultrasound entropy imaging can describe the risk of MetS for individuals with NAFLD and is superior to the conventional ASQ technique.

Effect of ultrasound frequency on the Nakagami statistics of human liver tissues.

The analysis of the backscattered statistics using the Nakagami parameter is an emerging ultrasound technique for assessing hepatic steatosis and fibrosis. Previous studies indicated that the echo amplitude distribution of a normal liver follows the Rayleigh distribution (the Nakagami parameter m is close to 1). However, using different frequencies may change the backscattered statistics of normal livers. This study explored the frequency dependence of the backscattered statistics in human livers and then discussed the sources of ultrasound scattering in the liver. A total of 30 healthy participants were enrolled to undergo a standard care ultrasound examination on the liver, which is a natural model containing diffuse and coherent scatterers. The liver of each volunteer was scanned from the right intercostal view to obtain image raw data at different central frequencies ranging from 2 to 3.5 MHz. Phantoms with diffuse scatterers only were also made to perform ultrasound scanning using the same protocol for comparisons with clinical data. The Nakagami parameter-frequency correlation was evaluated using Pearson correlation analysis. The median and interquartile range of the Nakagami parameter obtained from livers was 1.00 (0.98-1.05) for 2 MHz, 0.93 (0.89-0.98) for 2.3 MHz, 0.87 (0.84-0.92) for 2.5 MHz, 0.82 (0.77-0.88) for 3.3 MHz, and 0.81 (0.76-0.88) for 3.5 MHz. The Nakagami parameter decreased with the increasing central frequency (r = -0.67, p < 0.0001). However, the effect of ultrasound frequency on the statistical distribution of the backscattered envelopes was not found in the phantom results (r = -0.147, p = 0.0727). The current results demonstrated that the backscattered statistics of normal livers is frequency-dependent. Moreover, the coherent scatterers may be the primary factor to dominate the frequency dependence of the backscattered statistics in a liver.

Detecting changes in ultrasound backscattered statistics by using Nakagami parameters: Comparisons of moment-based and maximum likelihood estimators.

The Nakagami distribution is an approximation useful to the statistics of ultrasound backscattered signals for tissue characterization. Various estimators may affect the Nakagami parameter in the detection of changes in backscattered statistics. In particular, the moment-based estimator (MBE) and maximum likelihood estimator (MLE) are two primary methods used to estimate the Nakagami parameters of ultrasound signals. This study explored the effects of the MBE and different MLE approximations on Nakagami parameter estimations. Ultrasound backscattered signals of different scatterer number densities were generated using a simulation model, and phantom experiments and measurements of human liver tissues were also conducted to acquire real backscattered echoes. Envelope signals were employed to estimate the Nakagami parameters by using the MBE, first- and second-order approximations of MLE (MLE1 and MLE2, respectively), and Greenwood approximation (MLEgw) for comparisons. The simulation results demonstrated that, compared with the MBE and MLE1, the MLE2 and MLEgw enabled more stable parameter estimations with small sample sizes. Notably, the required data length of the envelope signal was 3.6 times the pulse length. The phantom and tissue measurement results also showed that the Nakagami parameters estimated using the MLE2 and MLEgw could simultaneously differentiate various scatterer concentrations with lower standard deviations and reliably reflect physical meanings associated with the backscattered statistics. Therefore, the MLE2 and MLEgw are suggested as estimators for the development of Nakagami-based methodologies for ultrasound tissue characterization.

Acoustic structure quantification by using ultrasound Nakagami imaging for assessing liver fibrosis.

Acoustic structure quantification (ASQ) is a recently developed technique widely used for detecting liver fibrosis. Ultrasound Nakagami parametric imaging based on the Nakagami distribution has been widely used to model echo amplitude distribution for tissue characterization. We explored the feasibility of using ultrasound Nakagami imaging as a model-based ASQ technique for assessing liver fibrosis. Standard ultrasound examinations were performed on 19 healthy volunteers and 91 patients with chronic hepatitis B and C (n = 110). Liver biopsy and ultrasound Nakagami imaging analysis were conducted to compare the METAVIR score and Nakagami parameter. The diagnostic value of ultrasound Nakagami imaging was evaluated using receiver operating characteristic (ROC) curves. The Nakagami parameter obtained through ultrasound Nakagami imaging decreased with an increase in the METAVIR score (p < 0.0001), representing an increase in the extent of pre-Rayleigh statistics for echo amplitude distribution. The area under the ROC curve (AUROC) was 0.88 for the diagnosis of any degree of fibrosis (≥F1), whereas it was 0.84, 0.69, and 0.67 for ≥F2, ≥F3, and ≥F4, respectively. Ultrasound Nakagami imaging is a model-based ASQ technique that can be beneficial for the clinical diagnosis of early liver fibrosis.

Ultrasound window-modulated compounding Nakagami imaging: Resolution improvement and computational acceleration for liver characterization.

Ultrasound Nakagami imaging is an attractive method for visualizing changes in envelope statistics. Window-modulated compounding (WMC) Nakagami imaging was reported to improve image smoothness. The sliding window technique is typically used for constructing ultrasound parametric and Nakagami images. Using a large window overlap ratio may improve the WMC Nakagami image resolution but reduces computational efficiency. Therefore, the objectives of this study include: (i) exploring the effects of the window overlap ratio on the resolution and smoothness of WMC Nakagami images; (ii) proposing a fast algorithm that is based on the convolution operator (FACO) to accelerate WMC Nakagami imaging. Computer simulations and preliminary clinical tests on liver fibrosis samples (n=48) were performed to validate the FACO-based WMC Nakagami imaging. The results demonstrated that the width of the autocorrelation function and the parameter distribution of the WMC Nakagami image reduce with the increase in the window overlap ratio. One-pixel shifting (i.e., sliding the window on the image data in steps of one pixel for parametric imaging) as the maximum overlap ratio significantly improves the WMC Nakagami image quality. Concurrently, the proposed FACO method combined with a computational platform that optimizes the matrix computation can accelerate WMC Nakagami imaging, allowing the detection of liver fibrosis-induced changes in envelope statistics. FACO-accelerated WMC Nakagami imaging is a new-generation Nakagami imaging technique with an improved image quality and fast computation.

Nanoscopic substructures of raft-mimetic liquid-ordered membrane domains revealed by high-speed single-particle tracking.

Lipid rafts are membrane nanodomains that facilitate important cell functions. Despite recent advances in identifying the biological significance of rafts, nature and regulation mechanism of rafts are largely unknown due to the difficulty of resolving dynamic molecular interaction of rafts at the nanoscale. Here, we investigate organization and single-molecule dynamics of rafts by monitoring lateral diffusion of single molecules in raft-containing reconstituted membranes supported on mica substrates. Using high-speed interferometric scattering (iSCAT) optical microscopy and small gold nanoparticles as labels, motion of single lipids is recorded via single-particle tracking (SPT) with nanometer spatial precision and microsecond temporal resolution. Processes of single molecules partitioning into and escaping from the raft-mimetic liquid-ordered (Lo) domains are directly visualized in a continuous manner with unprecedented clarity. Importantly, we observe subdiffusion of saturated lipids in the Lo domain in microsecond timescale, indicating the nanoscopic heterogeneous molecular arrangement of the Lo domain. Further analysis of the diffusion trajectory shows the presence of nano-subdomains of the Lo phase, as small as 10 nm, which transiently trap the lipids. Our results provide the first experimental evidence of non-uniform molecular organization of the Lo phase, giving a new view of how rafts recruit and confine molecules in cell membranes.

Shot-noise limited localization of single 20 nm gold particles with nanometer spatial precision within microseconds.

Single-particle tracking (SPT) is a powerful approach to investigate dynamics without ensemble average. Continuing effort has been made to track smaller particles with better spatial precision at higher speed. In this work, we demonstrate SPT of 20 nm gold nanoparticle (GNP) with 2 nm spatial precision up to 500 kHz by using microsecond interferometric scattering (µs-iSCAT) microscopy. The linear scattering signal from single GNPs is detected by a high-speed CMOS camera via interference. Through this homodyne detection, shot-noise limited sensitivity, and therefore optimal localization precision are achieved at high speed where considerable electronic noise is present. Using µs-iSCAT microscopy, we observe anomalous diffusion of GNPs labeled to lipid molecules in a supported bilayer membrane prepared on a glass substrate. The combination of nanometer spatial precision and microsecond temporal resolution provides the opportunity to study rapid motions of nano-objects on molecular scale with unprecedented clarity.

In vivo MR/optical imaging for gastrin releasing peptide receptor of prostate cancer tumor using Gd-TTDA-NP-BN-Cy5.5.

Magnetic resonance imaging (MRI) has become the leading imaging tool for providing fine anatomical and physiology details. Optical imaging is offering a sensitive and specific method for in vivo molecular imaging of targeting molecules. The goal of this study is to design, synthesize, and characterize a new target-specific dual contrast agent for MR and optical imaging. Hence, [Gd(TTDA-NP)(H(2)O)](2-) was prepared and characterized. In addition, an 8-amino acid Bombesin analogue (BN) peptide substrate, which can target prostate, breast, and colon cancer, was synthesized by solid-phase peptide synthesis and subsequently conjugated with [Gd(TTDA-NP)(H(2)O)](2-) to form BN conjugated Gd-TTDA-NP-BN. The water-exchange rate (k(ex)(298)) for [Gd(TTDA-NP)(H(2)O)](2-) (110×10(6)s(-1)) is significantly higher than that of [Gd(DTPA)(H(2)O)](2-) complex and the rotational correlation time (τ(R)) for [Gd(TTDA-NP)(H(2)O)](2-) (145ps) is also higher than those of [Gd(TTDA)(H(2)O)](2-) (104ps) and [Gd(DTPA)(H(2)O)](2-) (103ps). The Gd-TTDA-NP-BN shows remarkable high relaxivity (7.12mM(-1)s(-1)) comparing to that of [Gd(TTDA-NP)(H(2)O)](2-). The fluorescence studies showed that the Gd-TTDA-NP-BN could efficiently enter PC-3 cells. Additionally, the human cancer cells xenografts using Gd-TTDA-NP-BN-Cy5.5 as an optical imaging probe clearly visualized subcutaneous PC-3 tumor and demonstrated its targeting ability to the gastrin releasing peptide (GRP) receptor overexpression. Furthermore, the biodistribution studies demonstrated significantly high tumor uptake (25.97±1.07% ID/g) and high tumor-to-normal tissue ratios at one hour post-injection of Gd-TTDA-NP-BN-Cy5.5 in the animal model. These results suggest that the Gd-TTDA-NP-BN-Cy5.5 is a superior probe for in vivo optical imaging. Importantly, the MR imaging studies showed notable signal enhancement (44.9±4.2%) on the tumor, indicating a high level accumulation of the contrast agent within the PC-3 tumor sites. Hence, targeting of prostate cancer cells was observed under in vitro and in vivo MR imaging studies using Gd-TTDA-NP-BN contrast agent. We conclude that Gd-TTDA-NP-BN and Gd-TTDA-NP-BN-Cy5.5 can be potentially used as the contrast agents for targeting GRP receptor overexpressing cells and tumors.

Outer membrane protein STM3031 (Ail/OmpX-like protein) plays a key role in the ceftriaxone resistance of Salmonella enterica serovar Typhimurium.

Previously, the putative outer membrane protein STM3031 has been correlated with ceftriaxone resistance in Salmonella enterica serovar Typhimurium. In this study, this protein was almost undetectable in the ceftriaxone-susceptible strain 01-4, but its levels were increased in 01-4 isogenic strains for which MICs were higher. The stm3031 gene deletion mutant, R200(Deltastm3031), was generated and showed >64-fold lower ceftriaxone resistance than R200, supporting a key role for STM3031 in ceftriaxone resistance. To investigate which outer membrane protein(s) was associated with resistance, the outer membrane protein profiles of 01-4, R200, and R200(Deltastm3031) were compared proteomically. Nine proteins were identified as altered. The expression levels of AcrA, TolC, STM3031, STM1530, VacJ, and Psd in R200 were increased; those of OmpC, OmpD, and OmpW were decreased. The expression levels of OmpD, OmpW, STM1530, VacJ, and Psd, but not those of OmpC, AcrA, and TolC, in R200(Deltastm3031) were returned to the levels in strain 01-4. Furthermore, the genes' mRNA levels correlated with their protein levels when the three strains were compared. The detection of higher AcrB levels, linked to higher acrB, acrD, and acrF mRNA levels, in strain R200 than in strains 01-4 and R200(Deltastm3031) suggests that AcrB, AcrD, and AcrF participate in ceftriaxone resistance. Taken together with the location of STM3031 in the outer membrane, these results suggest that STM3031 plays a key role in ceftriaxone resistance, probably by reducing permeability via a decreased porin OmpD level and enhancing export via increased AcrD efflux pump activity.

A new isoquinolinone derivative with noble vasorelaxation activity.

The pharmacological effects of BDPBI (7-bromo-1,4-dihydro-2-phenyl-4,4-bis(4-pyridinylmethyl)2H-isoquinolin-3-one dihydrochloride) were tested on isolated endothelium-containing or denuded aorta of the guinea pig. BDPBI with the formula C(27)H(24)BrCl(2)N(3)O was synthesized starting with 3-isochromanone. In the endothelium-containing preparations of the aortic rings, phenylephrine (PHE; 10 micromol/l) elicited contracture and acetylcholine (ACh; 10 micromol/l) or BDPBI (0.01-10 micromol/l) elicited relaxation effects on the PHE-precontracted preparations. The BDPBI-elicited effect on the PHE-precontracted aortic rings was not altered in the presence of adrenergic blockers (propranolol or yohimbine; 1 micromol/l) or pretreated preparations with aspirin, indomethacin (10 micromol/l) or L-NAME (1 mmol/l). However, the relaxation effects of BDPBI were blocked if the preparations were pretreated with diphenhydramine (10 micromol/l) or chloropheniramine maleate (10 micromol/l). In contrast to lower concentrations of atropine (1 micromol/l), higher concentrations of atropine (30 micromol/l) did block the effects of BDPBI on the PHE-precontracted aortic rings. HTMT dimaleate (0.01-10 micromol/l), a histamine H(1) receptor agonist, also elicited relaxation effects on the PHE-precontracted preparation, and the effects were blocked if the preparations were pretreated with diphenhydramine or chloropheniramine maleate. On isolated denuded aorta of the guinea pig, BDPBI did not elicit relaxation effects on the PHE-precontracted aortic rings. These results demonstrated that the vasorelaxation effect of BDPBI on PHE-precontracted aortic rings is partly dependent on the activation of a histaminergic receptor from the vascular endothelium. We suggested that BDPBI would be an effective vasorelaxant for cardiovascular systems.