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Biomedical subjects

Y Pu

Publications and source records attributed to Y Pu.

At least 19 recordsLinked to original sources

Spectral polarization imaging of human prostate cancer tissue using a near-infrared receptor-targeted contrast agent.

The Cypate-Bombesin Peptide Analogue Conjugate (Cybesin) was used as a prostate tumor receptor-targeted contrast agent. The absorption and fluorescence spectra of Cybesin were measured and shown to exist in the NIR tissue "optical window". The spectral polarization imaging of Cybesin-stained prostate cancerous and normal tissues shows that prostate cancerous tissue takes-up more Cybesin than that of prostate normal tissue, making Cybesin a potential marker of prostate cancer.

Bombesin↗

Mobility of polymer chains confined at a free surface.

Dynamic secondary ion mass spectrometry was used to investigate the chain mobility of polystyrene (MW ranging from 4.3 to 957 kg/mol) at the free surface. The data show that the diffusion coefficient was reduced relative to the bulk value within a distance, d < or = 4R(g), from the surface and scaled as 1/N(2.5) at fixed d. These results are in excellent agreement with self-consistent field calculations of the surface segmental distribution and provide the first direct confirmation of various theoretical models that predict asymmetric segmental fluctuation which arises from surface induced orientation of polymer chains.

Journal Article↗

Application of the fluorescence resonance energy transfer method for studying the dynamics of caspase-3 activation during UV-induced apoptosis in living HeLa cells.

Activation of caspase-3 is a central event in apoptosis. We have developed a GFP-based FRET (fluorescence resonance energy transfer) probe that is highly sensitive to the activation of caspase-3 in intact living cells. This probe was constructed by fusing a CFP (cyan fluorescent protein) and a YFP (yellow fluorescent protein) with a specialized linker containing the caspase-3 cleavage sequence: DEVD. The linker design was optimized to produce a large FRET effect. Using purified protein, we observed a fivefold change in the fluorescence emission ratio when the probe was cleaved by caspase-3. To demonstrate the usefulness of this method, we introduced this FRET probe into HeLa cells by both transient and stable transfection. We observed that during UV-induced apoptosis, the activation of caspase-3 varied significantly between different cells; but once the caspase was activated, the enzyme within the cell became fully active within a few minutes. This technique will be highly useful for correlating the caspase-3 activation with other apoptotic events and for rapid-screening of potential drugs that may target the apoptotic process.

Apoptosis↗

Cytosolic Ca(2+) signal is involved in regulating UV-induced apoptosis in hela cells.

Results of recent studies using BAPTA/AM have raised a serious question on whether Ca(2+) signal is truly involved in regulating the progression of apoptosis. To resolve this question, we examined the differential effects of three different Ca(2+) signaling blockers (BAPTA/AM, membrane-impermeant BAPTA, and heparin) on UV-induced apoptosis in HeLa cells. We found that although the membrane-permeable form of BAPTA (i.e., BAPTA/AM) could not inhibit cell death, the membrane-impermeant form of BAPTA, loaded into the cytosol by electroporation, clearly protected cells from entering apoptosis. Furthermore, when we injected heparin to block Ca(2+) release from the endoplasmic reticulum (ER) to cytosol, apoptosis was greatly suppressed. These findings strongly suggest that elevation of cytosolic Ca(2+) is part of the signal that drives the progression of apoptosis. The negative result of BAPTA/AM is probably due to its dual effect on subcellular Ca(2+) distribution; besides suppressing the Ca(2+) elevation in cytosol, BAPTA/AM can also enter into the ER to reduce the free Ca(2+) level there. The depletion of Ca(2+) in ER is believed to stimulate apoptosis and thus would counterbalance the protection effect of BAPTA/AM in suppressing the cytosolic Ca(2+) elevation.

Apoptosis↗

Perfusion-weighted imaging of interictal hypoperfusion in temporal lobe epilepsy using FAIR-HASTE: comparison with H(2)(15)O PET measurements.

To detect perfusion abnormalities in areas of high magnetic susceptibility in the brain, an arterial spin-labeling MRI technique utilizing flow-sensitive alternating inversion recovery (FAIR) and half-Fourier single shot turbo spin-echo (HASTE) for spin preparation and image acquisition, respectively, was developed. It was initially tested in a functional study involving visual stimulation, and was able to detect significant activation with an increase (approximately 70%) in relative cerebral blood flow. Subsequently, it was applied in a clinical situation in eight patients with temporal lobe epilepsy (TLE). The perfusion-weighted images obtained showed no susceptibility artifacts even in the region of the inferior temporal lobe and were able to detect interictal hypoperfusion in TLE. The results were compared with those derived from H(2)(15)O PET perfusion imaging in each patient. A statistically significant correlation (r = 0.75, P < 0.05) was found between results acquired from these two modalities. Magn Reson Med 45:431-435, 2001.

Artifacts↗

Cooking oil fume-induced cytokine expression and oxidative stress in human lung epithelial cells.

Epidemiological studies have shown an association between exposure to indoor air pollution from Chinese-style cooking and risk of lung cancer among Chinese females. Several toxic substances have been identified in cooking oil fumes (COF) collected from heated rapeseed oil. In this study, we examined the biological effects of COF on CL3 human lung epithelial cells. Exposure to 200 microg/ml COF significantly reduced cell growth within 4 days. In addition, we examined the effect of COF on TGFbeta1, TGFbeta2, IL-6, IL-8, and IFN-gamma gene expressions with the RT-PCR method. We found that TGFbeta1 mRNA levels increased after exposure to 200 microg/ml COF for 24 h. Similarly, exposure to 10 microM benzo[a]pyrene or 100 nM 12-O-tetradecanoylphorbol-13-acetate increased TGFbeta1 mRNA levels at 24 h. The mRNA levels of TGFbeta2, IL-6, IL-8, and IFN-gamma did not increase after treatment with COF, benzo[a]pyrene, or 12-O-tetradecanoylphorbol-13-acetate. COF-induced TGFbeta1 production was confirmed by quantification of TGFbeta1 in conditioned medium with enzyme-linked immunosorbent assay. Exposure to 200 microg/ml COF significantly increased TGFbeta1 secretion in a time-dependent and dose-dependent manner. It has been demonstrated that reactive oxygen intermediates induce TGFbeta1 gene expression. When CL3 cells were exposed to 200 microg/ml COF for 15 min, there was an increase in intracellular peroxide formation with the dichlorofluorescein method. Furthermore, treatment with 200 microg/ml COF for 12 h also significantly induced lipid peroxidation in CL3 cells. Our results show that exposure to COF inhibits cell growth, increases TGFbeta1 secretion, and induces oxidative stress in CL3 lung epithelial cells. This suggests that TGFbeta1 and oxidative stress play a role in the biological effects of COF on lung epithelial cells.

Air Pollution, Indoor↗

Cerebral hemodynamic response in Chinese (first) and English (second) language processing revealed by event-related functional MRI.

Comparative functional neuroimaging studies using the block design paradigm have previously demonstrated that there are no significant differences in the location of areas of cerebral activation when native Chinese speakers independently process single words or sentences in both the Chinese (first) and English (second) languages. While it has also been documented that significant domains of brain response include the inferior to middle left frontal lobe, the latency, amplitude and duration of the associated hemodynamic changes during isolated neural processing of Chinese and English languages still remain unknown. The aim of this study, therefore, was to examine the characteristics of the hemodynamic alterations in the above-mentioned regions with event-related functional MRI (ER-fMRI) when native Chinese speakers performed verb generation tasks in both the Chinese (first) and English (second) languages. Our results demonstrate the presence of a similar neural activity-induced hemodynamic response in the inferior to middle left frontal lobe during both tasks. Further, there were also no statistically significant differences among the variables that described the hemodynamic response curves. These findings strongly imply that the underlying neural mechanism for Chinese (first) and English (second) language processing may be similar in native Chinese speakers.

Adult↗

Temporal relationship between cytochrome c release and mitochondrial swelling during UV-induced apoptosis in living HeLa cells.

During apoptosis, cytochrome c is released from mitochondria to the cytosol to activate a caspase cascade, which commits the cell to the death process. It has been proposed that the release of cytochrome c is caused by a swelling of the mitochondrial matrix triggered by the apoptotic stimuli. To test this theory, we measured directly the dynamic re-distribution of green fluorescence protein (GFP)-tagged cytochrome c and morphological change of mitochondria within living HeLa cells during u.v.-induced apoptosis. We observed that mitochondria did not swell when cytochrome c was released from mitochondria to cytosol during apoptosis. Instead, mitochondria swelled to spherical shapes within 10 minutes of cytochrome c release. This finding strongly suggests that cytochrome c release in apoptosis was not caused by mitochondrial swelling. This conclusion was further supported in two separated experiments using an immunostaining method and carbonyl cyanide m-chlorophenyl-hydrazone (CCCP) treatment. In addition, we found evidence that cytochrome c was also released before mitochondrial swelling in apoptosis induced by other cell death-inducing treatments, including tumor necrosis factor (TNF) and actinomycin D.

Amino Acid Chloromethyl Ketones↗

[Rat keratinocyte primary cultures based on conductive polypyrrole primary cell culture technique].

Orthogonal experimental design has been applied to study the suitable cell culture medium and condition of conductive polypyrrole on rat keratinocyte cell cultures. The results showed that one thousand milliliters of complete minimum essential medium(CMEM) which contained 900 ml minimum essential medium(MEM), 10% fetal bovine serum(FBS), 1% NEAA, 6 ml insulin and 10 mg hydrocortisone was fit for keratinocyte cell culture. And the conductive polypyrrole obtained with 10 mA current density for 100 second switching times, given 100 mV electrical stimulation voltage for 2 hours, would stimulate the growth of rat keratinocyte cell in vitro cultures. The cell density under culture conditions described above was 1.30 times as high as that under routine culture conditions.

Animals↗

[Detection of aflatoxin-albumin adducts in human serum and its application].

Exposure to aflatoxin B1 (AFB1) is one of the major risk factors in the etiology of human hepatocellular carcinoma (HCC). The development of highly sensitive and specific methods for detecting AFB1 and its macromolecular biomarkers are critical for identifying individuals at risk for hepatocellular carcinoma. The detection of aflatoxin-albumin adducts (AFB-ALB) in serum has been used to investigate the relationship between aflatoxin exposure and HCC. A more sensitive and reliable method to detect AFB-ALB in serum was modified by separating serum albumin with Microcon-50 microconcentrators in stead of using precipitation. The method could separate serum albumin quickly and make protein digested in the same container. Thus, the step for transferring samples was reduced and recovery rate was improved. The AFB-ALB adducts were then determined by radioimmunoassay. The AFB-ALB adducts of human serum samples from Qidong, Jiangsu Province, and Fusui, Guangxi Zhuang Autonomous Region of the People's Republic of China were detected. The results indicated that the exposure of AFB1 is common and the positive rate for AFB-ALB adducts in serum of these populations is high.

Aflatoxin B1↗

Tamm-Horsfall protein binds to type 1 fimbriated Escherichia coli and prevents E. coli from binding to uroplakin Ia and Ib receptors.

The adherence of uropathogenic Escherichia coli to the urothelial surface, a critical first step in the pathogenesis of urinary tract infection (UTI), is controlled by three key elements: E. coli adhesins, host receptors, and host defense mechanisms. Although much has been learned about E. coli adhesins and their urothelial receptors, little is known about the role of host defense in the adherence process. Here we show that Tamm-Horsfall protein (THP) is the principal urinary protein that binds specifically to type 1 fimbriated E. coli, the main cause of UTI. The binding was highly specific and saturable and could be inhibited by d-mannose and abolished by endoglycosidase H treatment of THP, suggesting that the binding is mediated by the high-mannose moieties of THP. It is species-conserved, occurring in both human and mouse THPs. In addition, the binding to THP was much greater with an E. coli strain bearing a phenotypic variant of the type 1 fimbrial FimH adhesin characteristic of those prevalent in UTI isolates compared with the one prevalent in isolates from the large intestine of healthy individuals. Finally, a physiological concentration of THP completely abolished the binding of type 1 fimbriated E. coli to uroplakins Ia and Ib, two putative urothelial receptors for type 1 fimbriae. These results establish, on a functional level, that THP contains conserved high-mannose moieties capable of specific interaction with type 1 fimbriae and strongly suggest that this major urinary glycoprotein is a key urinary anti-adherence factor serving to prevent type 1 fimbriated E. coli from binding to the urothelial receptors.

Animals↗

Shear modulation force microscopy study of near surface glass transition temperatures

We report results of glass transition (T(g)) measurements for polymer thin films using atomic force microscopy (AFM). The AFM mode, shear modulation force microscopy (SMFM), involves measuring the temperature-dependent shear force on a tip modulated parallel to the sample surface. Using this method we have measured the surface T(g) of thin (17-500 nm) polymer films and found that T(g) is independent of film thickness (t>17 nm), strength of substrate interactions, or even presence of substrate.

Journal Article↗

Detection of the brain response during a cognitive task using perfusion-based event-related functional MRI.

Event-related (ER) fMRI has evoked great interest due to the ability to depict the dynamic features of human brain function during various cognitive tasks. Thus far, all cognitive ER-fMRI studies have been based on blood oxygenation level-dependent (BOLD) contrast techniques. Compared with BOLD-based fMRI techniques, perfusion-based fMRI is able to localize the region of neuronal activity more accurately. This report demonstrates, for the first time, the detection of the brain response to a cognitive task using high temporal resolution perfusion-based ER-fMRI. An English verb generation task was used in this study. Results show that perfusion-based ER-fMRI accurately depicts the activation in Broca's area. Average changes in regional relative cerebral blood flow reached a maximum value of 30.7% at approximately 6.5 s after the start of stimulation and returned to 10% of the maximum value at approximately 12.8 s. Our results show that perfusion-based ER-fMRI is a useful tool for cognitive neuroscience studies, providing comparable temporal resolution and better localization of brain function than BOLD ER-fMRI.

Adult↗

Rapid delineation of humic and non-humic organic matter fractions in water.

Dissolved organic matter (DOM) in water is often characterized by aggregate parameters like dissolved organic carbon (DOC). DOM from conventional surface water treatment plant in Northern New Jersey was isolated and fractionated using resin adsorption chromatography into six different fractions, which were operationally categorized as hydrophobic acid, hydrophobic neutral, hydrophobic base, hydrophilic acid, hydrophilic neutral and hydrophilic base. The spectral fluorescent signatures (SFS) technique was developed for the quantitative identification of the six fractions by post-processing analysis that includes a statistical model. The SFS is the total sum of emission spectra of a sample at different excitation wavelengths, recorded as a matrix of fluorescent intensity in coordinates of excitation and emission wavelengths, in a definite spectral window. High sensitivity and rapid identification and quantification of DOM fractions are among the main features of the technique. Since hydrophobic and hydrophilic substances are considered more humic and non-humid in nature, respectively, the technique provided an opportunity to rapidly delineate source waters in terms of such categories.

Adsorption↗

In vivo fMRI demonstration of hypothalamic function following intraperitoneal glucose administration in a rat model.

The hypothalamic functional MRI (fMRI) response in an animal model was studied following energy intake. Six fasted (12 h) Sprague-Dawley (SD) rats were administered an intraperitoneal injection of glucose (0.72 grams/kg body weight), while a mid-sagittal slice through the hypothalamus was continuously imaged for 60 min using a conventional T2*-weighted gradient-echo sequence. All rats demonstrated a significant acute transient decrease in the fMRI signal intensity (mean: 3.4%) in the hypothalamic region within 12-16 min after intraperitoneal glucose injection. The SD rat may be a suitable model for future fMRI studies of the hypothalamus involving the administration of exogenous nutrients and medications.

Animals↗

Comparison of the temporal response in perfusion and BOLD-based event-related functional MRI.

Event-related functional MRI (ER-fMRI) based on both blood oxygen level-dependent (BOLD) contrast and perfusion contrast has been recently developed to study human brain activation due to brief stimulation. In this report, both BOLD- and perfusion-based ER-fMRI were directly compared using repeated single-trial, short visual stimulation (1 sec) in six human volunteers. The results show that the cerebral blood flow change reached a maximum approximately 1 sec earlier than the BOLD signal change (4.2 +/- 0.2 sec vs. 5.1 +/- 0.2 sec after the stimulation, P < 0.05). The full width at half maximum of the hemodynamic response measured by perfusion was not significantly different from that measured with BOLD (5.1 +/- 0.6 sec vs. 5.9 +/- 0.6 sec). A positive linear correlation was found between the maximum perfusion and maximum BOLD signal changes (r = 0. 77, P = 0.07).

Cerebrovascular Circulation↗

The human red nucleus and lateral cerebellum in supporting roles for sensory information processing.

A functional MRI study compared activation in the red nucleus to that in the lateral cerebellar dentate nucleus during passive and active tactile discrimination tasks. The study pursued recent neuroimaging results suggesting that the cerebellum may be more associated with sensory processing than with the control of movement for its own sake. Because the red nucleus interacts closely with the cerebellum, the possibility was examined that activity in red nucleus might also be driven by the requirement for tactile sensory processing with the fingers rather than by finger movement alone. The red and dentate nuclei were about 300% more active (a combination of activation areas and intensities) during passive (non-motor) tactile stimulation when discrimination was required than when it was not. Thus, the red nucleus was activated by purely sensory stimuli even in the absence of the opportunity to coordinate finger movements or to use the sensory cues to guide movement. The red and dentate nuclei were about 70% more active during active tactile tasks when discrimination was required than when it was not (i.e., for simple finger movements alone). Thus, the red nucleus was most active when the fingers were being used for tactile sensory discrimination. In both the passive and active tactile tasks, the observed activation had a contralateralized pattern, with stronger activation in the left red nucleus and right dentate nucleus. Significant covariation was observed between activity in the red nucleus and the contralateral dentate during the discrimination tasks and no significant correlation between the red nucleus and the contralateral dentate activity was detected during the two non-discrimination tasks. The observed interregional covariance and contralateralized activation patterns suggest strong functional connectivity during tactile discrimination tasks. Overall, the pattern of findings suggests that the activity in the red nucleus, as in the lateral cerebellum, is more driven by the requirements for sensory processing than by motor coordination per se.

Adult↗

Increased detectability of alpha brain glutamate/glutamine in neonatal hypoxic-ischemic encephalopathy.

BACKGROUND AND PURPOSE: Proton MR spectroscopy (MRS) detectability of brain glutamate/glutamine (Glx) is increased in hypoxic-ischemic insults and is implicated in the neuronal injury and death that follows. Our aim was to correlate the proton MRS detectability of alpha-CH protons of Glx (alpha-Glx) with the Sarnat stage of neonatal hypoxic-ischemic encephalopathy (HIE). METHODS: Initial and follow-up proton MRS studies at 1.9 T were performed in 28 neonates aged 1 to 7 days (seven healthy control subjects and 21 with HIE: 10 mild, nine moderate, and two severe) and in 12 neonates aged 13 to 17 days (12 with HIE: eight mild, three moderate, and one severe), respectively. Both point-resolved spectroscopy (PRESS) and stimulated-echo acquisition mode (STEAM) sequences were used. The spectral volume of interest was in the basal ganglia, thalami, and adjoining regions. The detectability of alpha-Glx was assessed by two different parameters: the detection rate of the alpha-Glx peak and the peak-area ratio of alpha-Glx to creatine and phosphocreatine. RESULTS: On both the initial and follow-up PRESS studies, all the neonates with moderate and severe HIE showed an alpha-Glx peak, compared with one healthy control subject in the initial study and one neonate with mild HIE in both the studies. They also demonstrated a significantly higher peak-area ratio of alpha-Glx/(creatine and phosphocreatine) on both the initial and follow-up studies. The peak-area ratios in neonates with HIE positively correlated with the Sarnat stage of HIE on both the initial and follow-up studies. Neonates with moderate and severe HIE also showed a consistently higher alpha-Glx peak on both the initial and follow-up studies with the STEAM sequence. CONCLUSION: Proton MRS detectability of alpha-Glx is increased in moderate and severe HIE and correlates with the Sarnat stage of HIE.

Brain↗