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Find video protocols related to scientific articles indexed in Pubmed.
[Rapid identification of potato cultivars using NIR-excited fluorescence and Raman spectroscopy].
Guang Pu Xue Yu Guang Pu Fen Xi
PUBLISHED: 09-12-2014
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Potato is one of the most important food in the world. Rapid and noninvasive identification of potato cultivars plays a important role in the better use of varieties. In this study, The identification ability of optical spectroscopy techniques, including near-infrared (NIR) Raman spectroscopy and NIR fluorescence spectroscopy, for invasive detection of potato cultivars was evaluated. A rapid NIR Raman spectroscopy system was applied to measure the composite Raman and NIR fluorescence spectroscopy of 3 different species of potatoes (98 samples in total) under 785 nm laser light excitation. Then pure Raman and NIR fluorescence spectroscopy were abstracted from the composite spectroscopy, respectively. At last, the partial least squares-discriminant analysis (PLS-DA) was utilized to analyze and classify Raman spectra of 3 different types of potatoes. All the samples were divided into two sets at random: the calibration set (74samples) and prediction set (24 samples), the model was validated using a leave-one-out, cross-validation method. The results showed that both the NIR-excited fluorescence spectra and pure Raman spectra could be used to identify three cultivars of potatoes. The fluorescence spectrum could distinguish the Favorita variety well (sensitivity: 1, specificity: 0.86 and accuracy: 0.92), but the result for Diamant (sensitivity: 0.75, specificity: 0.75 and accuracy: 0. 75) and Granola (sensitivity: 0.16, specificity: 0.89 and accuracy: 0.71) cultivars identification were a bit poorer. We demonstrated that Raman spectroscopy uncovered the main biochemical compositions contained in potato species, and provided a better classification sensitivity, specificity and accuracy (sensitivity: 1, specificity: 1 and accuracy: 1 for all 3 potato cultivars identification) among the three types of potatoes as compared to fluorescence spectroscopy.
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Multimodal nonlinear optical microscopic imaging provides new insights into acetowhitening mechanisms in live mammalian cells without labeling.
Biomed Opt Express
PUBLISHED: 09-01-2014
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We developed a multimodal nonlinear optical microscopy imaging (e.g., third-harmonic generation (THG) and two-photon excited fluorescence (TPEF)) platform based on a femtosecond laser pumped photonic crystal fiber to investigate the acetowhitening phenomenon induced by acetic acid in live mammalian cells without labeling. After treated by acetic acid with concentrations of higher than 0.2%, THG images show that light scattering is remarkably increased inside the nucleus and cytoplasm in cells. Co-localized TPEF and THG imaging on tryptophan and NADH in cells indicates that the change of scattering property is largely originating from the morphological change of metabolic proteins induced by acetic acids. Further TPEF imaging on NADH and FAD in cells confirms that this change is irreversible when acetic acid concentration is higher than 1.2%. These subcellular-level THG/TPEF imaging results reveal that the acetowhitening phenomenon is highly related with proteins involved in metabolic pathways in the nucleus and cytoplasm in live cells.
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Near-infrared Raman spectroscopy for assessing biochemical changes of cervical tissue associated with precarcinogenic transformation.
Analyst
PUBLISHED: 08-21-2014
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Raman spectroscopy measures the inelastically scattered light from tissue that is capable of identifying native tissue biochemical constituents and their changes associated with disease transformation. This study aims to characterize the Raman spectroscopic properties of cervical tissue associated with the multi-stage progression of cervical precarcinogenic sequence. A rapid-acquisition fiber-optic near-infrared (NIR) Raman diagnostic system was employed for tissue Raman spectral measurements at 785 nm excitation. A total of 68 Raman spectra (23 benign, 29 low-grade squamous intraepithelial lesions (LSIL) and 16 high grade squamous intraepithelial lesions (HSIL)) were measured from 25 cervical tissue biopsy specimens, as confirmed by colposcopy-histopathology. The semi-quantitative biochemical modeling based on the major biochemicals (i.e., DNA, proteins (histone, collagen), lipid (triolein) and carbohydrates (glycogen)) in cervical tissue uncovers the stepwise accumulation of biomolecular changes associated with progressive cervical precarcinogenesis. Multi-class partial least squares-discriminant analysis (PLS-DA) together with leave-one tissue site-out, cross-validation yielded the diagnostic sensitivities of 95.7%, 82.8% and 81.3%; specificities of 100.0%, 92.3% and 88.5%,for discrimination among benign, LSIL and HSIL cervical tissues, respectively. This work suggests that the Raman spectral biomarkers have identified the potential to be used for monitoring the multi-stage cervical precarcinogenesis, forming the foundation of applying NIR Raman spectroscopy for the early diagnosis of cervical precancer in vivo at the molecular level.
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Long-Term Effects of Continuous Positive Airway Pressure on Blood Pressure and Prognosis in Hypertensive Patients with Coronary Heart Disease and Obstructive Sleep Apnea: A Randomized Controlled Trial.
Am. J. Hypertens.
PUBLISHED: 08-14-2014
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Obstructive sleep apnea (OSA) can result in hypertension and significantly increase cardiovascular morbidity and mortality. There are few reports on the long-term effects of continuous positive airway pressure (CPAP) on blood pressure in patients with uncontrolled hypertension with coronary heart disease (CHD) and OSA.
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Genotoxicity of tri- and hexavalent chromium compounds in vivo and their modes of action on DNA damage in vitro.
PLoS ONE
PUBLISHED: 08-11-2014
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Chromium occurs mostly in tri- and hexavalent states in the environment. Hexavalent chromium [Cr(VI)] compounds are extensively used in diverse industries, and trivalent chromium [Cr(III)] salts are used as micronutrients and dietary supplements. In the present work, we report that they both induce genetic mutations in yeast cells. They both also cause DNA damage in both yeast and Jurkat cells and the effect of Cr(III) is greater than that of Cr(VI). We further show that Cr(III) and Cr(VI) cause DNA damage through different mechanisms. Cr(VI) intercalates DNA and Cr(III) interferes base pair stacking. Based on our results, we conclude that Cr(III) can directly cause genotoxicity in vivo.
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Expression of sulfur uptake assimilation-related genes in response to cadmium, bensulfuron-methyl and their co-contamination in rice roots.
J Environ Sci (China)
PUBLISHED: 08-01-2014
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The responses of sulfur (S) uptake assimilation-related genes' expression in roots of two rice cultivars to cadmium (Cd), bensulfuron-methyl (BSM) and their co-contamination (Cd+BSM) were investigated by gene-chip microarray analysis and quantitative real-time PCR (QRT-PCR) technology. Treatments of Cd and Cd+BSM induced expression of sulfate transporter and permease genes, and promoted sulfate uptake in rice roots. Cd+BSM could alleviate Cd toxicity to cv. Fengmeizhan seedlings, probably due to Cd+BSM promoting greater S absorption by seedlings. Cd and Cd+BSM induced expression of sulfate assimilation-related genes, and thus activated the sulfur assimilation pathway. Cd and Cd+BSM induced expression of phytochelatin synthase and metallothionein genes, and induced expression of glutathione S-transferases (GSTs), glutathione synthase (GS) and S-containing antioxidation enzyme genes, which detoxified Cd(2+). It is suggested that (to cope with the toxicity of Cd, BSM and their co-contamination) the S uptake and assimilation pathway was activated in rice roots by increased expression of related genes, thus enhancing the supply of organic S for synthesis of Cd or BSM resistance-related substances.
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Effect of oleic acid on the levels of eight metal ions in human hepatoma SMMC-7721 cells.
Biol Trace Elem Res
PUBLISHED: 03-17-2014
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Nonalcoholic fatty liver disease (NAFLD) is the most common chronic liver disease. Its incidence is rising worldwide. However, no specific therapy has been shown to be effective in its treatment. In the present study, the in vitro NAFLD model was established in human SMMC-7721 cells by using oleic acid (OA). Then, content changes of eight cations, including sodium, magnesium, potassium, calcium, iron, copper, zinc, and manganese, were investigated in the experimental model. The results showed that OA induced a decrease in magnesium level, while an increase in iron one. Additionally, the supplementation of magnesium in the cell culture model was studied. It showed that magnesium ameliorated lipid accumulation induced by OA. Our results suggest that magnesium could decrease the risk of NAFLD and be used as a promising candidate for the treatment of NAFLD.
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Genome-wide analysis of radiation-induced mutations in rice (Oryza sativa L. ssp. indica).
Mol Biosyst
PUBLISHED: 01-24-2014
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Radiation has been efficiently used for rice germplasm innovation. However, the molecular mechanisms by which radiation induces mutations are still unclear. In this study, we performed whole genome sequencing to reveal the comprehensive mutations in rice treated with radiation. Red-1 (a rice rich in beneficial ingredients for human health) was derived from rice 9311 after ?-radiation. Solexa sequencing technology was applied to uncover the mutations. Compared with the 9311 genome, 9.19% of genome sequences were altered in the Red-1 genome. Among these alterations, there were 381,403 SNPs, 50,116 1-5 bp Indels, 1279 copy number variations, and 10,026 presence/absence variations. These alterations were located in 14,493 genes, the majority of which contained a kinase domain, leucine rich repeats, or Cyt_P450. Point mutations were the main type of variation in the Red-1 genome. Gene ontology clustering revealed that genes that are associated with cell components, binding function, catalytic activity and metabolic processes were susceptible to ?-radiation. It was also predicted that 8 mutated genes were involved in the biosynthetic pathways of beneficial products or pigment accumulation. We conclude that genome-wide analysis of mutations provides novel insights into the mechanisms by which radiation improves the beneficial ingredients in rice Red-1.
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Structural basis for hijacking CBF-? and CUL5 E3 ligase complex by HIV-1 Vif.
Nature
PUBLISHED: 01-10-2014
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The human immunodeficiency virus (HIV)-1 protein Vif has a central role in the neutralization of host innate defences by hijacking cellular proteasomal degradation pathways to subvert the antiviral activity of host restriction factors; however, the underlying mechanism by which Vif achieves this remains unclear. Here we report a crystal structure of the Vif-CBF-?-CUL5-ELOB-ELOC complex. The structure reveals that Vif, by means of two domains, organizes formation of the pentameric complex by interacting with CBF-?, CUL5 and ELOC. The larger domain (?/? domain) of Vif binds to the same side of CBF-? as RUNX1, indicating that Vif and RUNX1 are exclusive for CBF-? binding. Interactions of the smaller domain (?-domain) of Vif with ELOC and CUL5 are cooperative and mimic those of SOCS2 with the latter two proteins. A unique zinc-finger motif of Vif, which is located between the two Vif domains, makes no contacts with the other proteins but stabilizes the conformation of the ?-domain, which may be important for Vif-CUL5 interaction. Together, our data reveal the structural basis for Vif hijacking of the CBF-? and CUL5 E3 ligase complex, laying a foundation for rational design of novel anti-HIV drugs.
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Toona Sinensis and Moschus Decoction Induced Cell Cycle Arrest in Human Cervical Carcinoma HeLa Cells.
Evid Based Complement Alternat Med
PUBLISHED: 01-08-2014
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Toona sinensis and Moschus are two herb materials used in traditional Chinese medicine, most commonly for their various biological activities. In this study, we investigated the inhibitory effect of three decoctions from Toona sinensis, Moschus, and Toona sinensis and Moschus in combination on cell growth in several normal and cancer cell lines by cell viability assay. The results showed that the combined decoction exhibited the strongest anticancer effects, compared to two single decoctions. The observations indicated that the combined decoction did not induce cell apoptosis and autophagy in HeLa cells by fluorescence microscopy. Flow cytometry analysis revealed that the combined decoction arrested HeLa cell cycle progression in S-phase. After the decoction incubation, among 41 cell cycle related genes, eight were reduced, while five were increased in mRNA levels by real-time PCR assay. Western blotting showed that there were no apparent changes of protein levels of Cyclin E1, while P27 expression significantly declined and the levels of CDC7 and CDK7 obviously increased. The data suggest that the RB pathway is partially responsible for the decoction-induced S-phase cell cycle arrest in HeLa cells. Therefore, the combined decoction may have therapeutic potential as an anticancer formula for certain cancers.
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Electronic metal-support interactions in single-atom catalysts.
Angew. Chem. Int. Ed. Engl.
PUBLISHED: 01-06-2014
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The synthesis of single-atom catalysts and the control of the electronic properties of catalytic sites to arrive at superior catalysts is a major challenge in heterogeneous catalysis. A stable supported single-atom silver catalyst with a controllable electronic state was obtained by anti-Ostwald ripening. An electronic perturbation of the catalytic sites that is induced by a subtle change in the structure of the support has a strong influence on the intrinsic reactivity. The higher depletion of the 4d?electronic state of the silver atoms causes stronger electronic metal-support interactions, which leads to easier reducibility and higher catalytic activity. These results may improve our understanding of the nature of electronic metal-support interactions and lead to structure-activity correlations.
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Multivariate reference technique for quantitative analysis of fiber-optic tissue Raman spectroscopy.
Anal. Chem.
PUBLISHED: 11-12-2013
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We report a novel method making use of multivariate reference signals of fused silica and sapphire Raman signals generated from a ball-lens fiber-optic Raman probe for quantitative analysis of in vivo tissue Raman measurements in real time. Partial least-squares (PLS) regression modeling is applied to extract the characteristic internal reference Raman signals (e.g., shoulder of the prominent fused silica boson peak (?130 cm(-1)); distinct sapphire ball-lens peaks (380, 417, 646, and 751 cm(-1))) from the ball-lens fiber-optic Raman probe for quantitative analysis of fiber-optic Raman spectroscopy. To evaluate the analytical value of this novel multivariate reference technique, a rapid Raman spectroscopy system coupled with a ball-lens fiber-optic Raman probe is used for in vivo oral tissue Raman measurements (n = 25 subjects) under 785 nm laser excitation powers ranging from 5 to 65 mW. An accurate linear relationship (R(2) = 0.981) with a root-mean-square error of cross validation (RMSECV) of 2.5 mW can be obtained for predicting the laser excitation power changes based on a leave-one-subject-out cross-validation, which is superior to the normal univariate reference method (RMSE = 6.2 mW). A root-mean-square error of prediction (RMSEP) of 2.4 mW (R(2) = 0.985) can also be achieved for laser power prediction in real time when we applied the multivariate method independently on the five new subjects (n = 166 spectra). We further apply the multivariate reference technique for quantitative analysis of gelatin tissue phantoms that gives rise to an RMSEP of ?2.0% (R(2) = 0.998) independent of laser excitation power variations. This work demonstrates that multivariate reference technique can be advantageously used to monitor and correct the variations of laser excitation power and fiber coupling efficiency in situ for standardizing the tissue Raman intensity to realize quantitative analysis of tissue Raman measurements in vivo, which is particularly appealing in challenging Raman endoscopic applications.
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Comparative Studies of Tri- and Hexavalent Chromium Cytotoxicity and Their Effects on Oxidative State of Saccharomyces cerevisiae Cells.
Curr. Microbiol.
PUBLISHED: 08-24-2013
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Chromium is a significant mutagen and carcinogen in environment. We compared the effects of tri- and hexavalent chromium on cytotoxicity and oxidative stress in yeast. Cell growth was inhibited by Cr(3+) or Cr(6+), and Cr(6+) significantly increased the lethal rate compared with Cr(3+). Both Cr(3+) and Cr(6+) can enter into the yeast cells. The percent of propidium iodide permeable cells treated with Cr(3+) is almost five times as that treated with the same concentration of Cr(6+). Levels of TBARS, O2 (-), and carbonyl protein were significantly increased in both Cr(6+)- and Cr(3+)-treated cells in a concentration- and time-dependent manner. Moreover, the accumulation of these stress markers in Cr(6+)-treated cells was over the Cr(3+)-treated ones. The decreased GSH level and increased activity of GPx were observed after 300 ?M Cr(6+)-exposure compared with the untreated control, whereas there was no other change of GSH content in cells treated with Cr(3+) even at very high concentration. Exposure to both Cr(3+) and Cr(6+) resulted in the decrease of activities of SOD and catalase. Furthermore, the effect of Cr(6+) is stronger than that of Cr(3+). Null mutation sensitivity assay demonstrated that the gsh1 mutant was sensitive to Cr(6+) other than Cr(3+), the apn1 mutant is more sensitive to Cr(6+) than Cr(3+), and the rad1 mutant is sensitive to both Cr(6+) and Cr(3+). Therefore, Cr(3+) can be concluded to inhibit cell growth probably due to the damage of plasma membrane integrality in yeast. Although both tri- and hexavalent chromium can induce cytotoxicity and oxidative stress, the action mode of Cr(3+) is different from that of Cr(6+), and serious membrane damage caused by Cr(3+) is not the direct consequence of the increase of lipid peroxidation.
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Development of a beveled fiber-optic confocal Raman probe for enhancing in vivo epithelial tissue Raman measurements at endoscopy.
Opt Lett
PUBLISHED: 07-02-2013
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We report on the development of a beveled fiber-optic confocal Raman probe coupled with a ball lens for enhancing in vivo epithelial tissue Raman measurements at endoscopy. Our Monte Carlo simulations show that by selecting a proper fiber-ball lens distance and beveled angle of collection fibers, the confocal Raman probe design can be optimized for maximizing shallower tissue Raman measurements in epithelial tissue; in addition, the ratio of epithelium to stromal Raman photons collected using an optimized confocal Raman probe is approximately 19-fold higher than that using a volume-type Raman probe. Further experiments confirm that the confocal Raman endoscopic probe developed is in favor of probing superficial tissue Raman signals from a two-layer tissue phantom as well as esophagus tissue in vivo during endoscopy. This work suggests the great potential of applying the beveled fiber-optic confocal Raman probe for improving in vivo diagnosis of precancer occurring in epithelial tissue at endoscopy.
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Near-infrared-excited confocal Raman spectroscopy advances in vivo diagnosis of cervical precancer.
J Biomed Opt
PUBLISHED: 06-26-2013
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Raman spectroscopy is a unique optical technique that can probe the changes of vibrational modes of biomolecules associated with tissue premalignant transformation. This study evaluates the clinical utility of confocal Raman spectroscopy over near-infrared (NIR) autofluorescence (AF) spectroscopy and composite NIR AF/Raman spectroscopy for improving early diagnosis of cervical precancer in vivo at colposcopy. A rapid NIR Raman system coupled with a ball-lens fiber-optic confocal Raman probe was utilized for in vivo NIR AF/Raman spectral measurements of the cervix. A total of 1240 in vivo Raman spectra [normal (n=993), dysplasia (n=247)] were acquired from 84 cervical patients. Principal components analysis (PCA) and linear discriminant analysis (LDA) together with a leave-one-patient-out, cross-validation method were used to extract the diagnostic information associated with distinctive spectroscopic modalities. The diagnostic ability of confocal Raman spectroscopy was evaluated using the PCA-LDA model developed from the significant principal components (PCs) [i.e., PC4, 0.0023%; PC5, 0.00095%; PC8, 0.00022%, (p<0.05)], representing the primary tissue Raman features (e.g., 854, 937, 1095, 1253, 1311, 1445, and 1654 cm(-1)). Confocal Raman spectroscopy coupled with PCA-LDA modeling yielded the diagnostic accuracy of 84.1% (a sensitivity of 81.0% and a specificity of 87.1%) for in vivo discrimination of dysplastic cervix. The receiver operating characteristic curves further confirmed that the best classification was achieved using confocal Raman spectroscopy compared to the composite NIR AF/Raman spectroscopy or NIR AF spectroscopy alone. This study illustrates that confocal Raman spectroscopy has great potential to improve early diagnosis of cervical precancer in vivo during clinical colposcopy.
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[Research on biometric method of heart sound signal based on GMM].
Zhongguo Yi Liao Qi Xie Za Zhi
PUBLISHED: 06-20-2013
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Extraction of cepstral coefficients combined with Gaussian Mixture Model (GMM) is used to propose a biometric method based on heart sound signal.
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Crystal structure of NLRC4 reveals its autoinhibition mechanism.
Science
PUBLISHED: 06-13-2013
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Nucleotide-binding and oligomerization domain-like receptor (NLR) proteins oligomerize into multiprotein complexes termed inflammasomes when activated. Their autoinhibition mechanism remains poorly defined. Here, we report the crystal structure of mouse NLRC4 in a closed form. The adenosine diphosphate-mediated interaction between the central nucleotide-binding domain (NBD) and the winged-helix domain (WHD) was critical for stabilizing the closed conformation of NLRC4. The helical domain HD2 repressively contacted a conserved and functionally important ?-helix of the NBD. The C-terminal leucine-rich repeat (LRR) domain is positioned to sterically occlude one side of the NBD domain and consequently sequester NLRC4 in a monomeric state. Disruption of ADP-mediated NBD-WHD or NBD-HD2/NBD-LRR interactions resulted in constitutive activation of NLRC4. Together, our data reveal the NBD-organized cooperative autoinhibition mechanism of NLRC4 and provide insight into its activation.
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Non-invasive analysis of hormonal variations and effect of postmenopausal Vagifem treatment on women using in vivo high wavenumber confocal Raman spectroscopy.
Analyst
PUBLISHED: 06-03-2013
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This study aims to evaluate the feasibility of applying high wavenumber (HW) confocal Raman spectroscopy for non-invasive assessment of menopause-related hormonal changes in the cervix as well as for determining the effect of Vagifem(®) treatment on postmenopausal women with atrophic cervix. A rapid HW confocal Raman spectroscopy system coupled with a ball lens fiber-optic Raman probe was utilized for in vivo cervical tissue Raman measurements at 785 nm excitation. A total of 164 in vivo HW Raman spectra (premenopausal (n = 104), postmenopausal-prevagifem (n = 34), postmenopausal-postvagifem (n = 26)) were measured from the normal cervix of 26 patients undergoing colposcopy. We established the biochemical basis of premenopausal, postmenopausal-prevagifem and postmenopausal-postvagifem cervix using semiquantitative biomolecular modeling derived from Raman-active biochemicals (i.e., lipids, proteins and water) that play a critical role in HW Raman spectral changes associated with the menopausal process. The diagnostic algorithms developed based on partial least squares-discriminant analysis (PLS-DA) together with leave-one patient-out, cross-validation yielded the diagnostic sensitivities of 88.5%, 91.2% and 88.5%, and specificities of 91.7%, 90.8% and 99.3%, respectively, for non-invasive in vivo discrimination among premenopausal, postmenopausal-prevagifem and postmenopausal-postvagifem cervix. This work demonstrates for the first time that HW confocal Raman spectroscopy in conjunction with biomolecular modeling can be a powerful diagnostic tool for identifying hormone/menopause-related variations in the native squamous epithelium of normal cervix, as well as for assessing the effect of Vagifem treatment on postmenopausal atrophic cervix in vivo during clinical colposcopic inspections.
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Circularly polarized coherent anti-Stokes Raman scattering microscopy.
Opt Lett
PUBLISHED: 04-19-2013
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We demonstrate circularly polarized coherent anti-Stokes Raman scattering (CP-CARS) microscopy that significantly suppresses the nonresonant background for high-contrast vibrational imaging. Circularly polarized pump and Stokes fields with opposite handedness are used to excite CARS signal. In this case, theoretically the nonresonant CARS signal and resonant CARS signal from isotropic media will completely vanish, while the resonant CARS signal from anisotropic structures can still exist. This allows CARS imaging of anisotropic samples with enhanced resonant contrast. Furthermore, we performed CP-CARS imaging on fibroin fibers from silkworm silk, and the results confirmed its effectiveness in background suppression.
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Ionic Extraction of a Novel Nano-sized Bioactive Glass Enhances Differentiation and Mineralization of Human Dental Pulp Cells.
J Endod
PUBLISHED: 03-21-2013
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This study aimed to investigate the effects of a novel nano-sized 58S bioactive glass (nano-58S BG) on the odontogenic differentiation and mineralization of human dental pulp cells (hDPCs) in vitro.
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Development of a multiplexing fingerprint and high wavenumber Raman spectroscopy technique for real-time in vivo tissue Raman measurements at endoscopy.
J Biomed Opt
PUBLISHED: 03-02-2013
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We report on the development of a novel multiplexing Raman spectroscopy technique using a single laser light together with a volume phase holographic (VPH) grating that simultaneously acquires both fingerprint (FP) and high wavenumber (HW) tissue Raman spectra at endoscopy. We utilize a customized VPH dual-transmission grating, which disperses the incident Raman scattered light vertically onto two separate segments (i.e., -150 to 1950??cm?¹; 1750 to 3600??cm?¹) of a charge-coupled device camera. We demonstrate that the multiplexing Raman technique can acquire high quality in vivo tissue Raman spectra ranging from 800 to 3600??cm?¹ within 1.0 s with a spectral resolution of 3 to 6??cm?¹ during clinical endoscopy. The rapid multiplexing Raman spectroscopy technique covering both FP and HW ranges developed in this work has potential for improving in vivo tissue diagnosis and characterization at endoscopy.
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Schnurri-3 regulates ERK downstream of WNT signaling in osteoblasts.
J. Clin. Invest.
PUBLISHED: 02-19-2013
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Mice deficient in Schnurri-3 (SHN3; also known as HIVEP3) display increased bone formation, but harnessing this observation for therapeutic benefit requires an improved understanding of how SHN3 functions in osteoblasts. Here we identified SHN3 as a dampener of ERK activity that functions in part downstream of WNT signaling in osteoblasts. A D-domain motif within SHN3 mediated the interaction with and inhibition of ERK activity and osteoblast differentiation, and knockin of a mutation in Shn3 that abolishes this interaction resulted in aberrant ERK activation and consequent osteoblast hyperactivity in vivo. Additionally, in vivo genetic interaction studies demonstrated that crossing to Lrp5(-/-) mice partially rescued the osteosclerotic phenotype of Shn3(-/-) mice; mechanistically, this corresponded to the ability of SHN3 to inhibit ERK-mediated suppression of GSK3?. Inducible knockdown of Shn3 in adult mice resulted in a high-bone mass phenotype, providing evidence that transient blockade of these pathways in adults holds promise as a therapy for osteoporosis.
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The relationship between serum calcium level, blood lipids, and blood pressure in hypertensive and normotensive subjects who come from a normal university in east of China.
Biol Trace Elem Res
PUBLISHED: 01-31-2013
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Previous studies revealed that low calcium intake is related to high prevalence of cardiovascular diseases such as hypertension. However, the relationship between serum calcium and blood pressure was unclear. The prevalence of hypertension is high in China. Thus, the aim of this study was to evaluate the serum calcium level between hypertensive and normotensive groups and to investigate the correlation between serum calcium, blood pressure, and blood lipid parameters. A total of 1,135 adult subjects participated in this study and were divide into two study groups: a hypertensive group (n?=?316) who had 140 mmHg or higher in systolic blood pressure (SBP) or 90 mmHg or higher in diastolic blood pressure (DBP) and an age- and sex-matched normotensive group (n?=?819, 120 mmHg or less SBP and 80 mmHg or less DBP). Our results indicate a significant trend for men (60 years old or older) in the direction of decreasing blood pressure with increasing serum calcium level, but no trend for women was indicated. In the normotensive group, a significant positive correlation was found between DBP and total cholesterol (P?
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Proteomic changes induced by podophyllotoxin in human cervical carcinoma HeLa cells.
Am. J. Chin. Med.
PUBLISHED: 01-23-2013
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Podophyllotoxin, a kind of lignan extracted from the Podophyllum plant, has been shown to inhibit the growth of various carcinoma cells. However, the molecular mechanism remains unclear. In this study, the inhibition of cell growth and changes in protein expression induced by podophyllotoxin were investigated in human cervical carcinoma HeLa cells. Our results demonstrate that Podophyllotoxin inhibits HeLa cell growth and induces apoptosis. By using proteomic techniques, seven proteins were found to be significantly regulated by podophyllotoxin compared to the untreated control; among them, four were down-regulated and three were up-regulated. All of the seven proteins were identified with peptide mass fingerprinting using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) after in-gel trypsin digestion. Five of these proteins are involved in protein metabolism, and the other two play roles in cell communication and signaling transduction pathways. It is suggested that the effect of podophyllotoxin on the growth of tumor cells is significantly related to the metabolism-associated proteins.
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A functional variomics tool for discovering drug-resistance genes and drug targets.
Cell Rep
PUBLISHED: 01-16-2013
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Comprehensive discovery of genetic mechanisms of drug resistance and identification of in vivo drug targets represent significant challenges. Here we present a functional variomics technology in the model organism Saccharomyces cerevisiae. This tool analyzes numerous genetic variants and effectively tackles both problems simultaneously. Using this tool, we discovered almost all genes that, due to mutations or modest overexpression, confer resistance to rapamycin, cycloheximide, and amphotericin B. Most significant among the resistance genes were drug targets, including multiple targets of a given drug. With amphotericin B, we discovered the highly conserved membrane protein Pmp3 as a potent resistance factor and a possible target. Widespread application of this tool should allow rapid identification of conserved resistance mechanisms and targets of many more compounds. New genes and alleles that confer resistance to other stresses can also be discovered. Similar tools in other systems, such as human cell lines, will also be useful.
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Serum calcium levels are not associated with coronary heart disease.
Vasc Health Risk Manag
PUBLISHED: 01-01-2013
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Numerous studies have reported that low calcium intake is related to a higher prevalence of cardiovascular disease. However, the relationship between serum calcium and coronary heart disease is unclear. The purpose of this study was to compare serum calcium levels in patients with coronary heart disease and those in healthy individuals.
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Assessment of liver steatosis and fibrosis in rats using integrated coherent anti-Stokes Raman scattering and multiphoton imaging technique.
J Biomed Opt
PUBLISHED: 11-25-2011
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We report the implementation of a unique integrated coherent anti-Stokes Raman scattering (CARS), second-harmonic generation (SHG), and two-photon excitation fluorescence (TPEF) microscopy imaging technique developed for label-free monitoring of the progression of liver steatosis and fibrosis generated in a bile duct ligation (BDL) rat model. Among the 21 adult rats used in this study, 18 rats were performed with BDL surgery and sacrificed each week from weeks 1 to 6 (n = 3 per week), respectively; whereas 3 rats as control were sacrificed at week 0. Colocalized imaging of the aggregated hepatic fats, collagen fibrils, and hepatocyte morphologies in liver tissue is realized by using the integrated CARS, SHG, and TPEF technique. The results show that there are significant accumulations of hepatic lipid droplets and collagen fibrils associated with severe hepatocyte necrosis in BDL rat liver as compared to a normal liver tissue. The volume of normal hepatocytes keeps decreasing and the fiber collagen content in BDL rat liver follows a growing trend until week 6; whereas the hepatic fat content reaches a maximum in week 4 and then appears to stop growing in week 6, indicating that liver steatosis and fibrosis induced in a BDL rat liver model may develop at different rates. This work demonstrates that the integrated CARS and multiphoton microscopy imaging technique has the potential to provide an effective means for early diagnosis and detection of liver steatosis and fibrosis without labeling.
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Novel photocatalytic antibacterial activity of TiO2 microspheres exposing 100% reactive {111} facets.
Chem. Commun. (Camb.)
PUBLISHED: 11-03-2011
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TiO(2) microspheres constructed by well-crystallized faceted nanorods with high aspect ratios expose 100% photocatalytic reactive {111} facets on the spherical surface. The microspheres demonstrated excellent photocatalytic antibacterial activity towards Staphylococcus aureus due to effective suppression of photoinduced electron-hole pair recombination and active TiO(2)@?OH core-shell structure.
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Sampangine inhibits heme biosynthesis in both yeast and human.
Eukaryotic Cell
PUBLISHED: 09-09-2011
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The azaoxoaporphine alkaloid sampangine exhibits strong antiproliferation activity in various organisms. Previous studies suggested that it somehow affects heme metabolism and stimulates production of reactive oxygen species (ROS). In this study, we show that inhibition of heme biosynthesis is the primary mechanism of action by sampangine and that increases in the levels of reactive oxygen species are secondary to heme deficiency. We directly demonstrate that sampangine inhibits heme synthesis in the yeast Saccharomyces cerevisiae. It also causes accumulation of uroporphyrinogen and its decarboxylated derivatives, intermediate products of the heme biosynthesis pathway. Our results also suggest that sampangine likely works through an unusual mechanism-by hyperactivating uroporhyrinogen III synthase-to inhibit heme biosynthesis. We also show that the inhibitory effect of sampangine on heme synthesis is conserved in human cells. This study also reveals a surprising essential role for the interaction between the mitochondrial ATP synthase and the electron transport chain.
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In vivo diagnosis of cervical precancer using Raman spectroscopy and genetic algorithm techniques.
Analyst
PUBLISHED: 08-25-2011
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This study aimed to evaluate the clinical utility of applying near-infrared (NIR) Raman spectroscopy and genetic algorithm-partial least squares-discriminant analysis (GA-PLS-DA) to identify biomolecular changes of cervical tissues associated with dysplastic transformation during colposcopic examination. A total of 105 in vivo Raman spectra were measured from 57 cervical sites (35 normal and 22 precancer sites) of 29 patients recruited, in which 65 spectra were from normal sites, while 40 spectra were from cervical precancerous lesions (i.e., 7 low-grade CIN and 33 high-grade CIN). The GA feature selection technique incorporated with PLS was utilized to study the significant biochemical Raman bands for differentiation between normal and precancer cervical tissues. The GA-PLS-DA algorithm with double cross-validation (dCV) identified seven diagnostically significant Raman bands in the ranges of 925-935, 979-999, 1080-1090, 1240-1260, 1320-1340, 1400-1420, and 1625-1645 cm(-1) related to proteins, nucleic acids and lipids in tissue, and yielded a diagnostic accuracy of 82.9% (sensitivity of 72.5% (29/40) and specificity of 89.2% (58/65)) for precancer detection. The results of this exploratory study suggest that Raman spectroscopy in conjunction with GA-PLS-DA and dCV methods has the potential to provide clinically significant discrimination between normal and precancer cervical tissues at the molecular level.
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Diagnosis of early stage nasopharyngeal carcinoma using ultraviolet autofluorescence excitation-emission matrix spectroscopy and parallel factor analysis.
Analyst
PUBLISHED: 08-04-2011
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We report the diagnostic ability of ultraviolet (UV)-excited autofluorescence (AF) excitation-emission matrix (EEM) spectroscopy associated with parallel factor (PARAFAC) analysis for differentiating cancer from normal nasopharyngeal tissue. A bifurcated fiber-optic probe coupled with an EEM system was used to acquire tissue AF EEMs using excitation wavelengths between 260 and 400 nm, and emission collection between 280 and 500 nm. A total of 152 AF EEM landscapes were acquired from 13 normal and 16 nasopharyngeal carcinoma (NPC) thawed ex vivo tissue samples from 23 patients. PARAFAC was introduced for curve resolution of individual AF EEM landscapes associated with the endogenous tissue constituents. The significant factors were further fed to a support vector machine (SVM) and cross-validated to construct diagnostic algorithms. Both the EEM intensity landscapes and the PARAFAC model revealed tryptophan, collagen, and elastin to be the three major endogenous fluorophores responsible for the AF signal from normal and NPC tissues. The EEM intensity distribution and PARAFAC factors suggest an increase of tryptophan and a decrease of collagen and elastin in NPC tissues compared to the normal. The classification results obtained from the PARAFAC-SVM modeling yielded a diagnostic accuracy of 94.7% (sensitivity of 95.0% (76/80); specificity of 94.4% (68/72)) for normal and NPC tissue differentiation. This study suggests that UV-excited AF EEM spectroscopy integrated with PARAFAC algorithms has the potential to provide clinical diagnostics of early onset and progression of NPC.
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Near-infrared autofluorescence spectroscopy for in vivo identification of hyperplastic and adenomatous polyps in the colon.
Biosens Bioelectron
PUBLISHED: 07-21-2011
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This study reports the implementation of an endoscope-based near-infrared (NIR) autofluorescence (AF) spectroscopy technique for in vivo differentiation of normal, hyperplastic and adenomatous colonic polyps during clinical colonoscopic examination. A total of 198 in vivo NIR AF spectra in the range of 810-1050 nm were acquired from colonic tissues (normal (n=116); hyperplastic (n=48); and adenomatous polyps (n=34)) of 96 patients undergoing colonoscopic screening. Significant differences (p<0.001, one-way analysis of variance (ANOVA)) in in vivo NIR AF intensity among normal, hyperplastic, and adenomatous polyps are observed. Multivariate statistical techniques, including principal components analysis (PCA) and linear discriminate analysis (LDA) together with the leave-one tissue site-out, cross-validation, were used to develop diagnostic algorithms for distinguishing adenomatous polyps from normal and hyperplastic colonic polyps based on NIR AF spectral features. The PCA-LDA modeling on in vivo colonic NIR AF dataset yields diagnostic sensitivities of 83.6%, 77.1%, and 88.2%; and specificities of 96.3%, 88.0%, and 92.1%, respectively, for classification of normal, hyperplastic and adenomatous colonic polyps. This work suggests that NIR AF spectroscopy associated with PCA-LDA algorithms has potential for in vivo diagnosis and detection of colonic precancer at colonoscopy.
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In vivo diagnosis of colonic precancer and cancer using near-infrared autofluorescence spectroscopy and biochemical modeling.
J Biomed Opt
PUBLISHED: 07-05-2011
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The aim of this study is to evaluate the biochemical foundation and clinical capability of an image-guided near-infrared (NIR) autofluorescence (AF) spectroscopy technique for in vivo diagnosis of colonic malignancies during clinical colonoscopy. A novel endoscopic fiber-optic AF system was utilized for in vivo NIR AF measurements at 785 nm excitation. A total of 263 in vivo NIR AF spectra of colonic tissues were measured from 100 patients, in which 164 spectra were from benign tissue (116 normal and 48 hyperplastic polyps), 34 spectra were from precancer (adenomatous polyps), and 65 spectra were from cancer. The non-negativity constrained least squares minimization biochemical modeling was explored to estimate the biochemical compositions of colonic tissue using nine basis reference spectra from the representative biochemicals (i.e., collagen I, elastin, ?-nicotinamide adenine dinucleotide, flavin adenine dinucleotide, L-tryptophan, hematoporphyrin, 4-pyridoxic acid, pyridoxal 5-phosphate, and water) associated with structural or cellular metabolic progression in colonic precancer and cancer. High-quality in vivo NIR AF spectra in the spectral range of 810 to 1000 nm were acquired from colonic benign, precancerous, and cancerous mucosa under white-light reflectance endoscopic imaging guidance. Partial least squares discriminant analysis, together with the leave-one tissue site-out, cross validation on in vivo NIR AF spectra yields diagnostic sensitivities of 85.4%, 76.5%, and 84.6%, and specificities of 89.9%, 93.4%, and 91.4%, respectively, for classification of benign, precancer, and cancer in the colon. This work demonstrates that image-guided NIR AF spectroscopy in conjunction with biochemical modeling has promising potential for improving in vivo detection and diagnosis of colonic precancer and cancer during clinical colonoscopic screening.
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Positive illusions and its association with cardiovascular functions.
Int J Psychophysiol
PUBLISHED: 04-12-2011
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The relationship between positive illusions (or self-enhancement) and cardiovascular functions was investigated using Asian samples in two studies. In phase 1 of Study 1, a generalized self-enhancement index was created for 241 participants using a paired word association memory task, a facial emotion recognition task, and a reading test. 122 participants subsequently volunteered for a second phase in this study where their ambulatory cardiovascular functions were measured throughout a single waking day. In Study 2, a priming procedure experimentally induced self-enhancement (n=35) and self-effacement (n=37) and the participants cardiovascular arousal and perceived control for a mental arithmetic task were measured. Self-enhancement predicted lower cardiovascular functions for both studies. In Study 1, self-enhancement assessed at phase 2 was a significant predictor while self-enhancement measured at phase 1 was not. In Study 2, the relationship between self-enhancement and vascular reactivity was partially mediated by perceived control. The findings indicate that the relationship between self-enhancement and cardiovascular stress response, which has implications for cardiovascular health, (i) is relevant for Asian populations, (ii) is not just correlational but potentially causal, and (iii) is partly mediated by an increase in perceived control for vascular reactivity.
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Characterizing variability in in vivo Raman spectra of different anatomical locations in the upper gastrointestinal tract toward cancer detection.
J Biomed Opt
PUBLISHED: 04-05-2011
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Raman spectroscopy is an optical vibrational technology capable of probing biomolecular changes of tissue associated with cancer transformation. This study aimed to characterize in vivo Raman spectroscopic properties of tissues belonging to different anatomical regions in the upper gastrointestinal (GI) tract and explore the implications for early detection of neoplastic lesions during clinical gastroscopy. A novel fiber-optic Raman endoscopy technique was utilized for real-time in vivo tissue Raman measurements of normal esophageal (distal, middle, and proximal), gastric (antrum, body, and cardia) as well as cancerous esophagous and gastric tissues from 107 patients who underwent endoscopic examinations. The non-negativity-constrained least squares minimization coupled with a reference database of Raman active biochemicals (i.e., actin, histones, collagen, DNA, and triolein) was employed for semiquantitative biomolecular modeling of tissue constituents in the upper GI. A total of 1189 in vivo Raman spectra were acquired from different locations in the upper GI. The Raman spectra among the distal, middle, and proximal sites of the esophagus showed no significant interanatomical variability. The interanatomical variability of Raman spectra among normal gastric tissue (antrum, body, and cardia) was subtle compared to cancerous tissue transformation, whereas biomolecular modeling revealed significant differences between the two organs, particularly in the gastroesophageal junction associated with proteins, DNA, and lipids. Cancerous tissues can be identified across interanatomical regions with accuracies of 89.3% [sensitivity of 92.6% (162?175); specificity of 88.6% (665?751)], and of 94.7% [sensitivity of 90.9% (30?33); specificity of 93.9% (216?230)] in the gastric and esophagus, respectively, using partial least squares-discriminant analysis together with the leave-one tissue site-out, cross validation. This work demonstrates that Raman endoscopy technique has promising clinical potential for real-time, in vivo diagnosis and detection of malignancies in the upper GI at the molecular level.
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Early detection of biomolecular changes in disrupted porcine cartilage using polarized Raman spectroscopy.
J Biomed Opt
PUBLISHED: 02-02-2011
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We evaluate the feasibility of applying polarized Raman spectroscopy in probing the early biochemical compositions and orientation changes in impacted porcine cartilage explants. We divide 100 fresh tibial cartilage explants into four groups: control (unimpacted) and 3 groups of single impact at 15, 20, and 25 MPa. Each group is examined for biochemical changes using Raman microscopy, cell viability changes using confocal fluorescence microscopy, and histological changes using the modified Mankin score. For the 15-MPa impact group, the modified Mankin score (p>0.05, n=15) and cell viability test (p>0.05, n=5) reveal no significant changes when compared to the control, but polarized Raman spectroscopy detects significant biochemical changes. A significant decrease in the parallel polarized intensity of the pyranose ring band at 1126 cm(-1) suggests a possible decrease in the glycoaminoglycan content in early cartilage damage (one-way analysis of variance with a post hoc Bonferonni test, p<0.05, n=10). For impacts greater than 15 MPa, cell viability and modified Mankin score are consistent with the changes in the observed polarized Raman signals. This suggests that the polarized Raman spectroscopy technique has potential for diagnosis and detection of early cartilage damage at the molecular level.
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Combining near-infrared-excited autofluorescence and Raman spectroscopy improves in vivo diagnosis of gastric cancer.
Biosens Bioelectron
PUBLISHED: 01-10-2011
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This study aims to evaluate the diagnostic utility of the combined near-infrared (NIR) autofluorescence (AF) and Raman spectroscopy for improving in vivo detection of gastric cancer at clinical gastroscopy. A rapid Raman endoscopic technique was employed for in vivo spectroscopic measurements of normal (n=1098) and cancer (n=140) gastric tissues from 81 gastric patients. The composite NIR AF and Raman spectra in the range of 800-1800 cm(-1) were analyzed using principal component analysis (PCA) and linear discriminant (LDA) to extract diagnostic information associated with distinctive spectroscopic processes of gastric malignancies. High quality in vivo composite NIR AF and Raman spectra can routinely be acquired from the gastric within 0.5s. The integrated intensity over the range of 800-1800 cm(-1) established the diagnostic implications (p=1.6E-14) of the change of NIR AF intensity associated with neoplastic transformation. PCA-LDA diagnostic modeling on the in vivo tissue NIR AF and Raman spectra acquired yielded a diagnostic accuracy of 92.2% (sensitivity of 97.9% and specificity of 91.5%) for identifying gastric cancer from normal tissue. The integration area under the receiver operating characteristic (ROC) curve using the combined NIR AF and Raman spectroscopy was 0.985, which is superior to either the Raman spectroscopy or NIR AF spectroscopy alone. This work demonstrates that the complementary Raman and NIR AF spectroscopy techniques can be integrated together for improving the in vivo diagnosis and detection of gastric cancer at endoscopy.
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Mapping the selection mechanisms by bacterial GEFs.
Virulence
PUBLISHED: 12-24-2010
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Mimicry of eukaryotic signaling enzymes is a common strategy used by bacterial pathogens to manipulate host cellular signaling. The E. coli type III effector protein Map belongs to a large family of bacterial virulence factors that activate host Rho GTPase signaling pathways through an unknown molecular mechanism. Our recent structural study, coupled with biochemical and functional assays, establishes that this family protein, including Map, IpgB1/2 and SifA/B secreted by E. coli, Shigella, and Salmonella respectively, acts as functional mimic of mammalian guanine nucleotide exchange factors (GEFs). Furthermore, we show that Map and its family members share a conserved mechanism with human Dbl GEFs for selection of various GTPase isoforms, revealing an evolutionary dynamic state of protein mimicry.
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Polarized near-infrared autofluorescence imaging combined with near-infrared diffuse reflectance imaging for improving colonic cancer detection.
Opt Express
PUBLISHED: 12-18-2010
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We evaluate the diagnostic feasibility of the integrated polarized near-infrared (NIR) autofluorescence (AF) and NIR diffuse reflectance (DR) imaging technique developed for colonic cancer detection. A total of 48 paired colonic tissue specimens (normal vs. cancer) were measured using the integrated NIR DR (850-1100 nm) and NIR AF imaging at the 785 nm laser excitation. The results showed that NIR AF intensities of cancer tissues are significantly lower than those of normal tissues (p<0.001, paired 2-sided Students t-test, n=48). NIR AF imaging under polarization conditions gives a higher diagnostic accuracy (of ~92-94%) compared to non-polarized NIR AF imaging or NIR DR imaging. Further, the ratio imaging of NIR DR to NIR AF with polarization provides the best diagnostic accuracy (of ~96%) among the NIR AF and NIR DR imaging techniques. This work suggests that the integrated NIR AF/DR imaging under polarization condition has the potential to improve the early diagnosis and detection of malignant lesions in the colon.
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Raman endoscopy for in vivo differentiation between benign and malignant ulcers in the stomach.
Analyst
PUBLISHED: 10-12-2010
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The aim of this study was to evaluate the clinical utility of an image-guided Raman endoscopy technique for in vivo differential diagnosis of benign and malignant ulcerous lesions in the stomach. A rapid-acquisition image-guided Raman endoscopy system with 785 nm laser excitation has been developed to acquire in vivo gastric tissue Raman spectra within 0.5 s during clinical gastroscopic examinations. A total of 1102 in vivo Raman spectra were acquired from 71 gastric patients, in which 924 Raman spectra were from normal tissue, 111 Raman spectra were from benign ulcers whereas 67 Raman spectra were from ulcerated adenocarcinoma. There were distinctive spectral differences in Raman spectra among normal mucosa, benign ulcers and malignant ulcers, particularly in the spectral ranges of 800-900, 1000-1100, 1245-1335, 1440-1450 and 1500-1800 cm(-1), which primarily contain signals related to proteins, DNA, lipids and blood. The malignant ulcerous lesions showed Raman signals to be mainly associated with abnormal nuclear activity and decrease in lipids as compared to benign ulcers. Partial least squares-discriminant analysis (PLS-DA) was employed to generate multi-class diagnostic algorithms for classification of Raman spectra of different gastric tissue types. The PLS-DA algorithms together with leave-one tissue site-out, cross validation technique yielded diagnostic sensitivities of 90.8%, 84.7%, 82.1%, and specificities of 93.8%, 94.5%, 95.3%, respectively, for classification of normal mucosa, benign and malignant ulcerous lesions in the stomach. This work demonstrates that image-guided Raman endoscopy technique associated with PLS-DA diagnostic algorithms has for the first time promising clinical potential for rapid, in vivo diagnosis and detection of malignant ulcerous gastric lesions at the molecular level.
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Integrated autofluorescence endoscopic imaging and point-wise spectroscopy for real-time in vivo tissue measurements.
J Biomed Opt
PUBLISHED: 08-31-2010
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We report on the development of an integrated point-wise spectroscopy and autofluorescence (AF) endoscopic imaging technique for real-time in vivo tissue measurements at endoscopy. We implement a unique point spectrum optical design to realize real-time AF imaging and AF or diffuse reflectance (DR) spectroscopy measurements from a small area of tissue of interest on the AF image. We demonstrate that both the AF image and the point-wise AF/DR spectra can be simultaneously acquired from the oral cavity in vivo within 0.1 s, suggesting the potential of the integrated spectroscopy and endoscopic imaging technique developed to facilitate in vivo tissue diagnosis and characterization at endoscopy.
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Triple-frequency symmetric subtraction scheme for nonresonant background suppression in coherent anti-Stokes Raman scattering (CARS) microscopy.
Opt Express
PUBLISHED: 08-20-2010
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We report a unique triple-frequency symmetric subtraction scheme to effectively remove the nonresonant background in coherent anti-Stokes Raman scattering (CARS) microscopy. Theoretical and experimental studies show that this unique scheme has an optimal performance for high contrast vibrational imaging, particularly useful when the resonant signal was larger than or comparable to the nonresonant background.
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Trivalent arsenic inhibits the functions of chaperonin complex.
Genetics
PUBLISHED: 07-26-2010
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The exact molecular mechanisms by which the environmental pollutant arsenic works in biological systems are not completely understood. Using an unbiased chemogenomics approach in Saccharomyces cerevisiae, we found that mutants of the chaperonin complex TRiC and the functionally related prefoldin complex are all hypersensitive to arsenic compared to a wild-type strain. In contrast, mutants with impaired ribosome functions were highly arsenic resistant. These observations led us to hypothesize that arsenic might inhibit TRiC function, required for folding of actin, tubulin, and other proteins postsynthesis. Consistent with this hypothesis, we found that arsenic treatment distorted morphology of both actin and microtubule filaments. Moreover, arsenic impaired substrate folding by both bovine and archaeal TRiC complexes in vitro. These results together indicate that TRiC is a conserved target of arsenic inhibition in various biological systems.
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The E3 ubiquitin ligase Wwp2 regulates craniofacial development through mono-ubiquitylation of Goosecoid.
Nat. Cell Biol.
PUBLISHED: 07-15-2010
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Craniofacial anomalies (CFAs) are the most frequently occurring human congenital disease, and a major cause of infant mortality and childhood morbidity. Although CFAs seems to arise from a combination of genetic factors and environmental influences, the underlying gene defects and pathophysiological mechanisms for most CFAs are currently unknown. Here we reveal a role for the E3 ubiquitin ligase Wwp2 in regulating craniofacial patterning. Mice deficient in Wwp2 develop malformations of the craniofacial region. Wwp2 is present in cartilage where its expression is controlled by Sox9. Our studies demonstrate that Wwp2 influences craniofacial patterning through its interactions with Goosecoid (Gsc), a paired-like homeobox transcription factor that has an important role in craniofacial development. We show that Wwp2-associated Gsc is a transcriptional activator of the key cartilage regulatory protein Sox6. Wwp2 interacts with Gsc to facilitate its mono-ubiquitylation, a post-translational modification required for optimal transcriptional activation of Gsc. Our results identify for the first time a physiological pathway regulated by Wwp2 in vivo, and also a unique non-proteolytic mechanism through which Wwp2 controls craniofacial development.
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In vivo early diagnosis of gastric dysplasia using narrow-band image-guided Raman endoscopy.
J Biomed Opt
PUBLISHED: 07-10-2010
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We first report on the implementation of a novel narrow-band image-guided Raman endoscopy technique for in vivo diagnosis of gastric dysplasia. High-quality in vivo Raman spectra can be acquired from normal and dysplastic gastric mucosal tissue within 0.5 sec under narrow-band image (NBI) guidance at gastroscopy. Significant differences are observed in in vivo Raman spectra between normal (n=54) and dysplastic (n=18) gastric tissue from 30 gastric patients, particularly in the spectral ranges of 825 to 950, 1000 to 1100, 1250 to 1500, and 1600 to 1800 cm(-1), which primarily contain signals related to proteins, nucleic acids, and lipids. The multivariate analysis [i.e., principal components analysis (PCA) and linear discriminant analysis (LDA)], together with the leave-one tissue site-out, cross validation on in vivo gastric Raman spectra yields a diagnostic sensitivity of 94.4% (1718) and specificity of 96.3% (5254) for distinction of gastric dysplastic tissue. This study suggests that narrowband image-guided Raman endoscopy associated with PCA-LDA diagnostic algorithms has potential for the noninvasive, in vivo early diagnosis and detection of gastric precancer during clinical gastroscopic examination.
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Effects of scatterers sizes on near-field coherent anti-Stokes Raman scattering under tightly focused radially and linearly polarized light excitation.
Opt Express
PUBLISHED: 07-01-2010
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We employ the finite-difference time-domain (FDTD) technique as a numerical approach to studying the effects of scatterers sizes on near-field coherent anti-Stokes Raman scattering (CARS) microscopy under tightly focused radially and linearly polarized light excitations. The FDTD results show that in a uniform medium (water), the full width at half maximum (FWHM) (transverse resolution) of radially polarized near-field CARS (RP-CARS) radiation is approximately 7.7% narrower than that of linearly polarized near-field CARS (LP-CARS) imaging, whereas the depth of focus (DOF) of RP-CARS radiation is 6.5% longer than LP-CARS. However, with the presence of scatterers in the uniform medium, both the FHWM and DOF of near-field RP-CARS radiation become much narrower compared to those of near-field LP-CARS radiation. In addition, the signal to nonresonant background ratio of near-field RP-CARS is significantly improved when the scatterers size is larger than a half wavelength of the pump light field. This work suggests that near-field CARS radiations are strongly influenced by the scatterers sizes in the medium; and near-field RP-CARS microscopy is superior to the near-field LP-CARS by providing both higher transverse and axial resolutions for three-dimensional molecular imaging of fine structures in biological systems.
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In vivo detection of epithelial neoplasia in the stomach using image-guided Raman endoscopy.
Biosens Bioelectron
PUBLISHED: 05-15-2010
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This study explores the utility of a novel image-guided Raman endoscopy technique for in vivo distinction of gastric cancer from normal tissue during clinical gastroscopy. The rapid-acquisition fiber-optic Raman endoscopy system developed was employed for in vivo gastric tissue Raman measurements at 785 nm laser excitation. A total of 1063 in vivo Raman spectra were acquired from 238 tissue sites of 67 gastric patients who underwent endoscopic ultrasound staging procedure, in which 934 Raman spectra were from 121 normal tissue sites whereas 129 Raman spectra were from 117 neoplastic gastric tissue sites. Gastric Raman spectra were fitted and reconstructed by using a linear combination of the eight basis reference spectra from the biochemicals (i.e., actin, albumin, collagen, DNA, histones, pepsinogen, phospholipids and triolein) in gastric tissue and also compared with the in vivo gastric Raman spectra measured. The resulting fit coefficients were further utilized through recursive partitioning techniques to develop diagnostic algorithms for gastric cancer diagnosis. High-quality in vivo Raman spectra in the range of 800-1800 cm(-1) can be acquired from gastric mucosa within 0.5s. The fit coefficients from albumin, nucleic acid, phospholipids and histones were found to be the most significant features for construction of the diagnostic model, giving rise to an overall accuracy of 93.7% (i.e., sensitivity of 94.0% (110/117) and specificity of 93.4% (113/121)) for in vivo discrimination of cancerous tissue from normal gastric tissue after the leave-one tissue site-out, cross-validation technique. This work demonstrates for the first time that image-guided Raman endoscopy technique has promising potential for the non-invasive, in vivo diagnosis and detection of gastric cancer at the molecular level.
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Autoprocessing and self-activation of the secreted protease CPAF in Chlamydia-infected cells.
Microb. Pathog.
PUBLISHED: 05-14-2010
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The Chlamydia-secreted protease/proteasome-like activity factor (CPAF) is synthesized as a proenzyme (proCPAF) and requires processing for proteolytic activity. Recent structural studies have further demonstrated that CPAF is a serine protease that can undergo autoprocessing and self-activation in a concentration-dependent manner in vitro. However, it is not known how CPAF is processed and activated during chlamydial infection. In the current study, we used a mutant CPAF designated as CPAF(E558A) that is deficient in processing by itself as a substrate to search for putative CPAF activation factor(s) in Chlamydia-infected cells. CPAF(E558A) was processed by the lysates made from Chlamydia-infected cells and the processing activity correlated with the presence of endogenous active CPAF in the fractionated lysate samples. CPAF produced in the Chlamydia-infected cells is required for processing the mutant CPAF(E558A) since the processing activity was removed by depletion with anti-CPAF but not control antibodies. Furthermore, a purified and activated wild type CPAF alone was sufficient for processing CPAF(E558A) and no other chlamydial proteases are required. Finally, fusion tag-induced oligomerization can lead to autoprocessing and self-activation of the wild type CPAF in mammalian cells. These observations together have demonstrated that CPAF undergoes autoprocessing and self-activation during chlamydial infection.
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Rapid near-infrared fluorescence excitation-emission matrix spectroscopy for multifluorophore characterization using an acousto-optic tunable filter technique.
J Biomed Opt
PUBLISHED: 05-13-2010
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We report on a novel acousto-optic tunable filter (AOTF)-based near-infrared (NIR) fluorescence excitation-emission matrix (EEM) spectroscopy technique for rapid multifluorophore characterization. We implement a unique light filtering module design by using cascaded AOTFs coupled with three orthogonally oriented polarizers to effectively remove the side-ripple artifacts of AOTFs as well as by using a pair of AOTFs coupled with two orthogonally oriented polarizers to improve detection efficiency for high-quality fluorescence EEM acquisitions. NIR fluorescence EEM spectroscopy (41 excitation wavelengths ranging from 550 to 950 nm in 10-nm increments; fluorescence emission from 570 to 1000 nm at 10-nm intervals) can be acquired from fluorescence dyes [e.g., diethylthiatricarbocyanine (DTTC) iodide, oxazine 750, and IR 140] within 10 s or even less, illustrating the potential of the AOTF-based NIR EEM technique developed for rapid multifluorophore analysis and characterization in biochemical and biomedical systems.
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Fiber-optic Raman probe couples ball lens for depth-selected Raman measurements of epithelial tissue.
Biomed Opt Express
PUBLISHED: 05-04-2010
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In this study, we present a fiber-optic ball lens Raman probe design for improving depth-selected Raman measurements of epithelial tissue. The Monte Carlo simulation results show that tissue Raman collection efficiency can be improved by properly selecting the refractive index and the diameter of the ball lens for the Raman probe design and the depth-selectivity of Raman measurements can also be improved by either increasing the refractive index or reducing the diameter of the ball lens. An appropriate arrangement of the Raman probe-tissue distance can also optimize the collection efficiency for depth-resolved Raman measurements. Experimental evaluation of a ball lens Raman probe design on a two-layer tissue phantom confirms the potential of the ball lens Raman probe design for efficient depth-selected measurement on epithelial tissue. This work suggests that the fiber-optic Raman probe coupled with a ball lens can facilitate the depth-selected Raman measurements of epithelial tissue, which may improve the diagnosis of epithelial precancer and early cancer at the molecular level.
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In vivo diagnosis of gastric cancer using Raman endoscopy and ant colony optimization techniques.
Int. J. Cancer
PUBLISHED: 05-03-2010
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This study aims to evaluate the clinical utility of image-guided Raman endoscopy for in vivo diagnosis of neoplastic lesions in the stomach at gastroscopy. A rapid-acquisition image-guided Raman endoscopy system with 785-nm excitation has been developed to acquire in vivo gastric tissue Raman spectra within 0.5 sec during clinical gastroscopic examinations. A total of 1,063 in vivo Raman spectra were acquired from 238 tissue sites of 67 gastric patients, in which 934 Raman spectra were from normal tissue whereas 129 Raman spectra were from neoplastic gastric tissue. The swarm intelligence-based algorithm (i.e., ant colony optimization (ACO) integrated with linear discriminant analysis (LDA)) was developed for spectral variables selection to identify the biochemical important Raman bands for differentiation between normal and neoplastic gastric tissue. The ACO-LDA algorithms together with the leave-one tissue site-out, cross validation method identified seven diagnostically important Raman bands in the regions of 850-875, 1,090-1,110, 1,120-1,130, 1,170-1,190, 1,320-1,340, 1,655-1,665 and 1,730-1,745 cm(-1) related to proteins, nucleic acids and lipids of tissue and provided a diagnostic sensitivity of 94.6% and specificity of 94.6% for distinction of gastric neoplasia. The predictive sensitivity of 89.3% and specificity of 97.8% were also achieved for an independent test validation dataset (20% of total dataset). This work demonstrates for the first time that the real-time image-guided Raman endoscopy associated with ACO-LDA diagnostic algorithms has potential for the noninvasive, in vivo diagnosis and detection of gastric neoplasia during clinical gastroscopy.
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Low-level detection of anti-cancer drug in blood plasma using microwave-treated gold-polystyrene beads as surface-enhanced Raman scattering substrates.
Biosens Bioelectron
PUBLISHED: 04-21-2010
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We report on a low-level detection of anti-cancer drug (paclitaxel) in blood plasma using microwave-treated gold (Au) film-polystyrene (PS) beads as surface-enhanced Raman scattering (SERS) substrates. SERS spectra of paclitaxel can be acquired within 10s at a concentration of 1×10(-8) M in blood plasma adsorbed on the modified Au-PS SERS substrates. The concentrations of paclitaxel ranging from 1.0×10(-8) to 1.0×10(-7) M in blood plasma can be well estimated with an accuracy of ?3.8×10(-9) M using SERS technique. This work shows the promise of applying microwave-modified Au-PS as SERS substrates for rapid and accurate monitoring of the anti-cancer drug in blood plasma for immediate treatment decisions during cancer therapy.
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High wavenumber Raman spectroscopy for in vivo detection of cervical dysplasia.
Anal. Chem.
PUBLISHED: 10-13-2009
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Raman spectroscopy is a vibrational spectroscopic technique capable of optically probing the biomolecular changes associated with neoplastic transformation. The purpose of this study was to apply near-infrared (NIR) Raman spectroscopy in the high wavenumber (HW) region (2800-3700 cm(-1)) for in vivo detection of cervical dysplasia. A rapid-acquisition NIR Raman spectroscopy system associated with a ball-lens fiber-optic Raman probe was developed for in vivo spectroscopic measurements at 785 nm excitation. A total of 92 in vivo HW Raman spectra (46 normal, 46 dysplasia) were acquired from 46 patients with Pap smear abnormalities of the cervix. Significant difference in Raman intensities of prominent Raman bands at 2850 and 2885 cm(-1) (CH(2) stretching of lipids), 2940 cm(-1) (CH(3) stretching of proteins), and the broad Raman band of water (peaking at 3400 cm(-1) in the 3100-3700 cm(-1) range) were observed in normal and dysplasia cervical tissue. The diagnostic algorithms based on principal components analysis and linear discriminant analysis together with the leave-one-patient-out cross-validation method on in vivo HW Raman spectra yielded a diagnostic sensitivity of 93.5% and specificity of 97.8% for dysplasia tissue identification. This study demonstrates for the first time that HW Raman spectroscopy has the potential for the noninvasive, in vivo diagnosis and detection of precancer of the cervix.
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Coherent anti-Stokes Raman scattering microscopy using tightly focused radially polarized light.
Opt Lett
PUBLISHED: 06-17-2009
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We report a radially polarized coherent anti-Stokes Raman scattering (RP-CARS) microscopy for facilitating longitudinally oriented molecules detection. We observe that under tight focusing of radially polarized pump and Stokes light fields with a high-NA objective, RP-CARS radiation from molecules oriented along the longitudinal direction is approximately threefold stronger than that using linearly polarized CARS (LP-CARS) technique. The lateral resolution of RP-CARS imaging can be improved by about 10% compared to the LP-CARS imaging. We demonstrate this RP-CARS technique by imaging the sectioned cottonwood leaf vascular bundles.
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Structural insights into host GTPase isoform selection by a family of bacterial GEF mimics.
Nat. Struct. Mol. Biol.
PUBLISHED: 05-06-2009
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The Escherichia coli type III effector Map belongs to a large family of bacterial virulence factors that activate host Rho GTPase signaling pathways through an unknown molecular mechanism. Here we report direct evidence that Map functions as a potent and selective guanine-nucleotide exchange factor (GEF) for Cdc42. The 2.3-A structure of the Map-Cdc42 complex revealed that Map mimics the GEF strategy of the mammalian Dbl family but has a three-dimensional architecture that is nearly identical to the bacterial GEF Salmonella spp. SopE. A comparative analysis between human and bacterial GEFs revealed a previously uncharacterized pairing mechanism between Map and the variable beta2-3 interswitch region of Cdc42. We propose a GTPase selection model that is experimentally validated by the preferential activation Rac1 and RhoA by the Shigella spp. effectors IpgB1 and IpgB2, respectively. These results significantly expand the repertoire of bacterial GEF mimics and unify a GEF selection mechanism for host GTPase substrates.
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Integrated Raman spectroscopy and trimodal wide-field imaging techniques for real-time in vivo tissue Raman measurements at endoscopy.
Opt Lett
PUBLISHED: 03-14-2009
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We report an integrated Raman spectroscopy and trimodal (white-light reflectance, autofluorescence, and narrow-band) imaging techniques for real-time in vivo tissue Raman measurements at endoscopy. A special 1.8 mm endoscopic Raman probe with filtering modules is developed, permitting effective elimination of interference of fluorescence background and silica Raman in fibers while maximizing tissue Raman collections. We demonstrate that high-quality in vivo Raman spectra of upper gastrointestinal tract can be acquired within 1 s or subseconds under the guidance of wide-field endoscopic imaging modalities, greatly facilitating the adoption of Raman spectroscopy into clinical research and practice during routine endoscopic inspections.
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Numerical study of effects of light polarization, scatterer sizes and orientations on near-field coherent anti-Stokes Raman scattering microscopy.
Opt Express
PUBLISHED: 02-17-2009
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We employ the finite-difference time-domain (FDTD) technique as a numerical approach to studying the effects of polarization, scatterers sizes and orientations on near-field coherent anti-Stokes Raman scattering (CARS) microscopy imaging. The results show that to acquire better image contrast and larger near-field CARS signals, the scatterers with diameters of less than three-eighths of the pump field wavelength (lambda(p)) are preferable to be oriented along the polarization direction of the excitation light fields. It is also found that when the scatterers sizes are smaller than half a wavelength of the pump field, the perpendicular polarization component of the induced near-field CARS radiations is strictly confined within the regions at the scatterer-water interface or the subsurface of scatterers, yielding a high image contrast (up to 200) with a spatial resolution of lambda(p)/16. This study indicates that perpendicular polarization components of near-field CARS microscopy could be used to uncover very fine structures of intra- and/or inter- cellular organelles in cells with nanoscale resolutions.
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Effects of emodin on the gene expression profiling of human breast carcinoma cells.
Cancer Detect. Prev.
PUBLISHED: 01-30-2009
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The mechanism of emodin-mediated cell apoptosis has been investigated extensively in many types of human cancer cells. Our previous study demonstrated that emodin induced apoptosis through the decrease of Bcl-2/Bax ratio and the increase of cytoplasm cytochrome c concentration in human breast cancer BCap-37 cells. However, emodins reaction to breast cancer cells remains elusive.
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Spectroscopic diagnosis of laryngeal carcinoma using near-infrared Raman spectroscopy and random recursive partitioning ensemble techniques.
Analyst
PUBLISHED: 01-07-2009
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In this work, we evaluated the diagnostic ability of near-infrared (NIR) Raman spectroscopy associated with the ensemble recursive partitioning algorithm based on random forests for identifying cancer from normal tissue in the larynx. A rapid-acquisition NIR Raman system was utilized for tissue Raman measurements at 785 nm excitation, and 50 human laryngeal tissue specimens (20 normal; 30 malignant tumors) were used for NIR Raman studies. The random forests method was introduced to develop effective diagnostic algorithms for classification of Raman spectra of different laryngeal tissues. High-quality Raman spectra in the range of 800-1800 cm(-1) can be acquired from laryngeal tissue within 5 seconds. Raman spectra differed significantly between normal and malignant laryngeal tissues. Classification results obtained from the random forests algorithm on tissue Raman spectra yielded a diagnostic sensitivity of 88.0% and specificity of 91.4% for laryngeal malignancy identification. The random forests technique also provided variables importance that facilitates correlation of significant Raman spectral features with cancer transformation. This study shows that NIR Raman spectroscopy in conjunction with random forests algorithm has a great potential for the rapid diagnosis and detection of malignant tumors in the larynx.
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Fiber-optic Raman spectroscopy probes gastric carcinogenesis in vivo at endoscopy.
J Biophotonics
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Intestinal-type gastric carcinogenesis is a complex multi-step disease, and early precursors (e.g. intestinal metaplasia (IM), dysplasia) can be very challenging to identify using conventional white-light endoscopic imaging. This study aims to assess the capability of Raman spectroscopy for multi-class elucidation of intestinal-type gastric carcinogenesis sequence in vivo for improving precancer detection at endoscopy. We employ a novel image-guided Raman endoscopy technique developed for in vivo gastric tissue Raman measurement within 0.5 s during clinical endoscopic examination. We have acquired a total of 1277 in vivo Raman spectra from 83 gastric patients associated with intestinal-type carcinogenesis. In vivo Raman spectroscopy integrated with semi-quantitative spectral modelling (e.g. DNA, lipids, glycoprotein, proteins and blood) reveals the progressive changes of biochemical constituents in gastric tissue associated with preneoplastic and neoplastic transformation (i.e., IM, dysplasia and adenocarcinoma). Multi-class probabilistic partial least squares-discriminant analysis (PLS-DA) diagnostic algorithms based on in vivo Raman spectra are able to identify normal mucosa with sensitivity of 75.88% and specificity of 87.21%; IM with sensitivity of 46.67% and specificity of 87.55%; dysplasia with sensitivity of 83.33%; specificity of 95.80%, and adenocarcinoma with sensitivity of 84.91% and specificity 95.57%, respectively. This work demonstrates that Raman spectroscopy is a sensitive biomolecular probe for monitoring intestinal-type gastric carcinogenesis to realize early diagnosis and detection of precancer and early gastric cancer in vivo during clinical endoscopic examination.
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Real-time Raman spectroscopy for in vivo, online gastric cancer diagnosis during clinical endoscopic examination.
J Biomed Opt
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Optical spectroscopic techniques including reflectance, fluorescence and Raman spectroscopy have shown promising potential for in vivo precancer and cancer diagnostics in a variety of organs. However, data-analysis has mostly been limited to post-processing and off-line algorithm development. In this work, we develop a fully automated on-line Raman spectral diagnostics framework integrated with a multimodal image-guided Raman technique for real-time in vivo cancer detection at endoscopy. A total of 2748 in vivo gastric tissue spectra (2465 normal and 283 cancer) were acquired from 305 patients recruited to construct a spectral database for diagnostic algorithms development. The novel diagnostic scheme developed implements on-line preprocessing, outlier detection based on principal component analysis statistics (i.e., Hotellings T2 and Q-residuals) for tissue Raman spectra verification as well as for organ specific probabilistic diagnostics using different diagnostic algorithms. Free-running optical diagnosis and processing time of < 0.5 s can be achieved, which is critical to realizing real-time in vivo tissue diagnostics during clinical endoscopic examination. The optimized partial least squares-discriminant analysis (PLS-DA) models based on the randomly resampled training database (80% for learning and 20% for testing) provide the diagnostic accuracy of 85.6% [95% confidence interval (CI): 82.9% to 88.2%] [sensitivity of 80.5% (95% CI: 71.4% to 89.6%) and specificity of 86.2% (95% CI: 83.6% to 88.7%)] for the detection of gastric cancer. The PLS-DA algorithms are further applied prospectively on 10 gastric patients at gastroscopy, achieving the predictive accuracy of 80.0% (60/75) [sensitivity of 90.0% (27/30) and specificity of 73.3% (33/45)] for in vivo diagnosis of gastric cancer. The receiver operating characteristics curves further confirmed the efficacy of Raman endoscopy together with PLS-DA algorithms for in vivo prospective diagnosis of gastric cancer. This work successfully moves biomedical Raman spectroscopic technique into real-time, on-line clinical cancer diagnosis, especially in routine endoscopic diagnostic applications.
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Discovering thiamine transporters as targets of chloroquine using a novel functional genomics strategy.
PLoS Genet.
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Chloroquine (CQ) and other quinoline-containing antimalarials are important drugs with many therapeutic benefits as well as adverse effects. However, the molecular targets underlying most such effects are largely unknown. By taking a novel functional genomics strategy, which employs a unique combination of genome-wide drug-gene synthetic lethality (DGSL), gene-gene synthetic lethality (GGSL), and dosage suppression (DS) screens in the model organism Saccharomyces cerevisiae and is thus termed SL/DS for simplicity, we found that CQ inhibits the thiamine transporters Thi7, Nrt1, and Thi72 in yeast. We first discovered a thi3? mutant as hypersensitive to CQ using a genome-wide DGSL analysis. Using genome-wide GGSL and DS screens, we then found that a thi7? mutation confers severe growth defect in the thi3? mutant and that THI7 overexpression suppresses CQ-hypersensitivity of this mutant. We subsequently showed that CQ inhibits the functions of Thi7 and its homologues Nrt1 and Thi72. In particular, the transporter activity of wild-type Thi7 but not a CQ-resistant mutant (Thi7(T287N)) was completely inhibited by the drug. Similar effects were also observed with other quinoline-containing antimalarials. In addition, CQ completely inhibited a human thiamine transporter (SLC19A3) expressed in yeast and significantly inhibited thiamine uptake in cultured human cell lines. Therefore, inhibition of thiamine uptake is a conserved mechanism of action of CQ. This study also demonstrated SL/DS as a uniquely effective methodology for discovering drug targets.
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A "smart" hollandite DeNO(x) catalyst: self-protection against alkali poisoning.
Angew. Chem. Int. Ed. Engl.
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Banish the villains to their own realm: biomass has gained widespread attention as a renewable energy source. However, commercial catalysts used in power plants (co-)fuelled by biomass are deactivated by the alkali-rich flue gas. In contrast, one of two types of active sites in a promising alkali-resistant hollandite catalyst traps alkali-metal ions to free up the catalytically active sites for the reduction of NO by NH(3).
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In vivo, real-time, transnasal, image-guided Raman endoscopy: defining spectral properties in the nasopharynx and larynx.
J Biomed Opt
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We report for the first time the implementation of transnasal, image-guided Raman endoscopy to directly assess Raman spectral properties of nasopharyngeal and laryngeal tissue in vivo during clinical endoscopic examinations. A rapid 785-nm excitation Raman endoscopy system, coupled with a miniaturized fiber-optic Raman probe, was utilized for real-time, in vivo Raman measurements of different anatomical locations in the head and neck. A total of 874 high-quality in vivo Raman spectra were successfully acquired from different anatomic locations of the nasopharynx and larynx [i.e., posterior nasopharynx (PN) (n=521), the fossa of Rosenmu?ller (FOR) (n=157), and true laryngeal vocal chords (LVC) (n=196)] in 23 normal subjects at transnasal endoscopy. Difference spectra and principal component analysis (PCA) were employed for tissue characterization, uncovering the tissue variability at the biomolecular level. The PCA-linear discriminant analysis (LDA) provides sensitivity of 77.0% and specificity of 89.2% for differentiation between PN and FOR, and sensitivity of 68.8% and specificity of 76.0% for distinguishing LVC and PN using the leave-one-subject-out, cross-validation method. This work demonstrates that transnasal, image-guided Raman endoscopy can be used to acquire in vivo Raman spectra from the nasopharynx and larynx in real time. Significant Raman spectral differences (p<0.05) identified as reflecting the distinct composition and morphology in the nasopharynx and larynx should be considered to be important parameters in the interpretation and rendering of diagnostic decision algorithms for in vivo tissue diagnosis and characterization in the head and neck.
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Simultaneous fingerprint and high-wavenumber confocal Raman spectroscopy enhances early detection of cervical precancer in vivo.
Anal. Chem.
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Raman spectroscopy is a vibrational spectroscopic technique capable of nondestructively probing endogenous biomolecules and their changes associated with dysplastic transformation in the tissue. The main objectives of this study are (i) to develop a simultaneous fingerprint (FP) and high-wavenumber (HW) confocal Raman spectroscopy and (ii) to investigate its diagnostic utility for improving in vivo diagnosis of cervical precancer (dysplasia). We have successfully developed an integrated FP/HW confocal Raman diagnostic system with a ball-lens Raman probe for simultaneous acquistion of FP/HW Raman signals of the cervix in vivo within 1 s. A total of 476 in vivo FP/HW Raman spectra (356 normal and 120 precancer) are acquired from 44 patients at clinical colposcopy. The distinctive Raman spectral differences between normal and dysplastic cervical tissue are observed at ~854, 937, 1001, 1095, 1253, 1313, 1445, 1654, 2946, and 3400 cm(-1) mainly related to proteins, lipids, glycogen, nucleic acids and water content in tissue. Multivariate diagnostic algorithms developed based on partial least-squares-discriminant analysis (PLS-DA) together with the leave-one-patient-out, cross-validation yield the diagnostic sensitivities of 84.2%, 76.7%, and 85.0%, respectively; specificities of 78.9%, 73.3%, and 81.7%, respectively; and overall diagnostic accuracies of 80.3%, 74.2%, and 82.6%, respectively, using FP, HW, and integrated FP/HW Raman spectroscopic techniques for in vivo diagnosis of cervical precancer. Receiver operating characteristic (ROC) analysis further confirms the best performance of the integrated FP/HW confocal Raman technique, compared to FP or HW Raman spectroscopy alone. This work demonstrates, for the first time, that the simultaneous FP/HW confocal Raman spectroscopy has the potential to be a clinically powerful tool for improving early diagnosis and detection of cervical precancer in vivo during clinical colposcopic examination.
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Optical diagnosis of laryngeal cancer using high wavenumber Raman spectroscopy.
Biosens Bioelectron
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We report the implementation of the transnasal image-guided high wavenumber (HW) Raman spectroscopy to differentiate tumor from normal laryngeal tissue at endoscopy. A rapid-acquisition Raman spectroscopy system coupled with a miniaturized fiber-optic Raman probe was utilized to realize real-time HW Raman (2800-3020 cm(-1)) measurements in the larynx. A total of 94 HW Raman spectra (22 normal sites, 72 tumor sites) were acquired from 39 patients who underwent laryngoscopic screening. Significant differences in Raman intensities of prominent Raman bands at 2845, 2880 and 2920 cm(-1) (CH(2) stretching of lipids), and 2940 cm(-1) (CH(3) stretching of proteins) were observed between normal and cancer laryngeal tissue. The diagnostic algorithms based on principal components analysis (PCA) and linear discriminant analysis (LDA) together with the leave-one subject-out, cross-validation method on HW Raman spectra yielded a diagnostic sensitivity of 90.3% (65/72) and specificity of 90.9% (20/22) for laryngeal cancer identification. This study demonstrates that HW Raman spectroscopy has the potential for the noninvasive, real-time diagnosis and detection of laryngeal cancer at the molecular level.
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Inhibition of type I insulin-like growth factor receptor tyrosine kinase by picropodophyllin induces apoptosis and cell cycle arrest in T lymphoblastic leukemia/lymphoma.
Leuk. Lymphoma
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Abstract It has been recently shown that IGF-IR contributes significantly to the survival of T lymphoblastic leukemia/lymphoma (T-LBL) cells, and it was therefore suggested that IGF-IR could represent a legitimate therapeutic target in this aggressive disease. Picropodphyllin (PPP) is a potent, selective inhibitor of IGF-IR that is currently used with notable success in clinical trials that include patients with aggressive types of epithelial tumors. In the present study, we tested the effects of PPP on Jurkat and Molt-3 cells; two prototype T-LBL cell lines. Our results demonstrate that PPP efficiently induced apoptotic cell death and cell cycle arrest of these two cells. These effects were attributable to alterations of downstream target proteins. By using proteomic analysis, 7 different proteins were found to be affected by PPP treatment of Jurkat cells. These proteins are involved in various aspects of cellular metabolism, cytoskeleton organization, and signal transduction pathways. The results suggest that PPP affects multiple signaling molecules and inhibits fundamental pathways that control cell growth and survival. Our study also provides novel evidence that PPP could be potentially utilized for the treatment of the aggressive T-LBL.
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JoVE Visualize is a tool created to match the last 5 years of PubMed publications to methods in JoVE's video library.

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In developing our video relationships, we compare around 5 million PubMed articles to our library of over 4,500 methods videos. In some cases the language used in the PubMed abstracts makes matching that content to a JoVE video difficult. In other cases, there happens not to be any content in our video library that is relevant to the topic of a given abstract. In these cases, our algorithms are trying their best to display videos with relevant content, which can sometimes result in matched videos with only a slight relation.