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Biomedical subjects

L Guan

Publications and source records attributed to L Guan.

At least 19 recordsLinked to original sources
PubMed2006

A three-phase, fully resorbable, polyester/calcium phosphate scaffold for bone tissue engineering: Evolution of scaffold design.

Bone tissue engineering strategies are fundamentally based upon porous scaffold materials that serve as a support for ingrowth of host cells and/or provide a substrate for exogenously delivered cells. Here we report the application of a surface calcium phosphate (CaP) mineral layer to a macroporous polymeric/CaP composite biomaterial, with a macroporous interconnectivity, and its subsequent in vivo evaluation in a rodent femoral defect. The application of the mineral layer eliminates the fibrous tissue encapsulation and foreign body giant cell response commonly seen at the interface of polymeric materials, yet retains the unique characteristics of the parent material as being macroporous, completely biodegradable and possessing a high degree of interconnectivity. This represents the third generation of this scaffold material, incorporating iterative changes to the scaffold design in response to both materials and biological design criteria to produce a material with enhanced in vitro and in vivo performance.

Absorbable Implants↗
PubMed2001

Helix packing in the lactose permease of Escherichia coli: localization of helix VI.

Plasmids encoding "split" lactose permease constructs with discontinuities in either the periplasmic loop between helices V and VI (N(5)/C(7)) or between helices VI and VII (N(6)/C(6)) were used to localize helix VI within the tertiary structure by site-directed thiol cross-linking. A total of 57 double-Cys pairs, with one Cys residue in helix VI and another in helix V or VIII, were studied with homobifunctional cross-linking agents. Significant cross-linking is observed between the periplasmic ends of helices V (position 158 or 161) and VI (position 170) with rigid 6 or 10 A reagents. Furthermore, the Cys residue at position 170 (helix VI) also cross-links to a Cys residue at either position 264 or 265 (helix VIII) with a 21 A cross-linking agent. The data indicate that helices V, VI and VIII are in close proximity at the periplasmic face of the membrane, with helix VI significantly closer to helix V. In addition, beta,D-galactopyranosyl 1-thio-beta,D-galactopyranoside induces a significant increase in cross-linking efficiency between helices VI and VIII and between helices V and VIII, with no significant change in cross-linking between helices V and VI.

Biological Transport↗
PubMed2001

Feature extraction of chromosomes from 3-D confocal microscope images.

An investigation of local energy surface detection integrated with neural network techniques for image segmentation is presented, as applied in the feature extraction of chromosomes from image datasets obtained using an experimental confocal microscope. Use of the confocal microscope enables biologists to observe dividing cells (living or preserved) within a three-dimensional (3-D) volume, that can be visualised from multiple aspects, allowing for increased structural insight. The Nomarski differential interference contrast mode used for imaging translucent specimens, such as chromosomes, produces images not suitable for volume rendering. Segmentation of the chromosomes from this data is, thus, necessary. A neural network based on competitive learning, known as Kohonen's self-organizing feature map (SOFM) was used to perform segmentation, using a collection of statistics or features defining the image. Our past investigation showed that standard features such as the localized mean and variance of pixel intensities provided reasonable extraction of objects such as mitotic chromosomes, but surface detail was only moderately resolved. In this current work, a biologically inspired feature known as local energy is investigated as an alternative image statistic based on phase congruency in the image. This, along with different combinations of other image statistics, is applied in a SOFM, producing 3-D images exhibiting vast improvement in the level of detail and clearly isolating the chromosomes from the background. Index Terms-DIC, differential interference contrast, feature extraction, feature space, image segmentation, local energy, Morlet wavelet, phase congruency, self organizing feature map, SOFM.

Algorithms↗
PubMed2001

Identification of essential charged residues in transmembrane segments of the multidrug transporter MexB of Pseudomonas aeruginosa.

The MexABM efflux pump exports structurally diverse xenobiotics, utilizing the proton electrochemical gradient to confer drug resistance on Pseudomonas aeruginosa. The MexB subunit traverses the inner membrane 12 times and has two, two, and one charged residues in putative transmembrane segments 2 (TMS-2), TMS-4, and TMS-10, respectively. All five residues were mutated, and MexB function was evaluated by determining the MICs of antibiotics and fluorescent dye efflux. Replacement of Lys342 with Ala, Arg, or Glu and Glu346 with Ala, Gln, or Asp in TMS-2 did not have a discernible effect. Ala, Asn, or Lys substitution for Asp407 in TMS-4, which is well conserved, led to loss of activity. Moreover, a mutant with Glu in place of Asp407 exhibited only marginal function, suggesting that the length of the side chain at this position is important. The only replacements for Asp408 in TMS-4 or Lys939 in TMS-10 that exhibited significant function were Glu and Arg, respectively, suggesting that the native charge at these positions is required. In addition, double neutral mutants or mutants in which the charged residues Asp407 and Lys939 or Asp408 and Lys939 were interchanged completely lost function. An Asp408-->Glu/Lys939-->Arg mutant retained significant activity, while an Asp407-->Glu/Lys939-->Arg mutant exhibited only marginal function. An Asp407-->Glu/Asp408-->Glu double mutant also lost activity, but significant function was restored by replacing Lys939 with Arg (Asp407-->Glu/Asp408-->Glu/Lys939-->Arg). Taken as a whole, the findings indicate that Asp407, Asp408, and Lys939 are functionally important and raise the possibility that Asp407, Asp408, and Lys939 may form a charge network between TMS-4 and TMS-10 that is important for proton translocation and/or energy coupling.

Amino Acid Sequence↗
PubMed2001

Evaluation of cytotoxic T-lymphocyte responses in human and nonhuman primate subjects infected with human immunodeficiency virus type 1 or simian/human immunodeficiency virus.

Cytotoxic T-lymphocyte (CTL) responses have been implicated as playing an important role in control of human immunodeficiency virus (HIV) infection. However, it is technically difficult to demonstrate CTL responses consistently in nonhuman primate and human subjects using traditional cytotoxicity assay methods. In this study, we systematically evaluated culture conditions that may affect the proliferation and expansion of CTL effector cells and presented a sensitive method for detection of cytotoxicity responses with bulk CTL cultures. We confirmed the sensitivity and specificity of this method by demonstration of vigorous CTL responses in a simian-HIV (SHIV)-infected rhesus macaque. The expansion of epitope-specific CTL effector cells was also measured quantitatively by CTL epitope-major histocompatibility complex tetramer complex staining. In addition, two new T-cell determinants in the SIV gag region are identified. Last, we showed the utility of this method for studying CTL responses in chimpanzee and human subjects.

Acquired Immunodeficiency Syndrome↗
PubMed2001

Spatiotemporal properties of an evoked population activity in rat sensory cortical slices.

We have examined the spatiotemporal properties of ensemble activity, an evoked all-or-none polysynaptic activity in rat neocortical slices. Ensemble activity occurred in cortical slices bathed in normal artificial cerebrospinal fluid (ACSF) and was evoked by a single electrical shock either to the white matter or directly to the cortical tissue. This activity was seen in slices of somatosensory and auditory cortices; in other cortical areas we have not been able to evoke it. The activity developed 10 to 250 ms poststimulus and lasted 280 +/- 120 ms in local field potential (LFP) recordings. Voltage-sensitive dye imaging showed that this activity was an area of activation 0.8 +/- 0.4 mm wide that propagated slowly (11.4 +/- 6.2 mm/s, n = 60, 6 animals) in the horizontal direction. Due to this propagation, the actual duration in the whole tissue may be longer (approximately 400 ms) than that recorded by a single LFP electrode. Ensemble activity produced a low-amplitude optical signal (7-14% of the interictal-like spikes in the same tissue), suggesting a moderate net depolarization of the population. These were very different from hyperexcitable (epileptiform) events in the same tissue that had about 10 times the optical signal amplitude and propagated at 125 +/- 24 mm/s (n = 21, 6 animals). On a global spatial scale (approximately 0.8 mm wide in layers II-III) ensemble activity had a smooth waveform in voltage-sensitive dye signals (population transmembrane potential). On a local scale, field potential recordings showed large fluctuations with complex oscillations and substantial trial-to-trial variation. This suggests that oscillations in cortical circuits occurred only in small clusters of correlated neurons. Ensemble activity was sensitive to the excitation-inhibition balance of the local network. Antagonists of N-methyl-D-aspartate, alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid, and GABAa receptors, and muscarinic agonists and other modest manipulations such as increasing bath concentration of Mg(2+) to 2.5-4 mM (normally at 2 mM), or K(+) to 5-7 mM (normally 3 mM), all significantly reduced the probability of evoking the activity. The metabotropic glutamate receptor agonist, aminocyclopentane-1,3-dicarboxylic acid, blocked the activity at a low concentration (10-15 microM), while the antagonist (R,S)-alpha-methyl-4-carboxyphenylglycine had no effect even at high concentration (240 microM). Our data suggest that locally organized neuronal clusters may play a role in the organization of oscillatory activities in the gamma band and may participate in cortical integration/amplification occurring on a scale of approximately 1 mm x 300 ms.

Animals↗
PubMed2001

Impact of movement and movement-related feedback on the lamprey central pattern generator for locomotion.

A semi-reduced, minimally restrained lamprey preparation was used to investigate the impact of movement and movement-related feedback during D-glutamate-induced locomotion. The preparation consisted of the trunk alone with the spinal cord exposed to the bathing solution. Two conditions were compared using electromyography or nerve recording: (i) muscle and spinal cord, (ii) spinal cord alone supported by the notochord. Compared with the isolated spinal cord, movement in the presence of muscle consistently and significantly increased the frequency of the motor output and reduced the phase delay among the segments. In moving preparations, coupling among the segments was reduced by two staggered hemisections to permit the strength and direction of intersegmental coupling to be estimated. The estimates revealed that movement increased the total intersegmental coupling strength and increased the proportion of the coupling that was descending over those of the isolated spinal cord. The effects on the phase and frequency of bursting can be explained in the light of the excitation evoked by bending that we have reported previously. Thus, we demonstrate that movement and movement-related feedback that arise from spinally induced motor patterns can alter the form of the movement and the functional coupling strength among the segments of the lamprey spinal cord.

Animals↗
PubMed2001

Metabolic changes in rats with photochemically induced cerebral infarction and the effects of batroxobin: a study by magnetic resonance imaging, 1H- and 31P-magnetic resonance spectroscopy.

Metabolic changes in rats with photochemically induced cerebral infarction and the effects of batroxobin were investigated 1, 3, 5 and 7 days after infarction by means of magnetic resonance imaging (MRI), 1H- and 31P-magnetic resonance spectroscopy (MRS). A region of T2 hyperintensity was observed in left temporal neocortex in infarction group and batroxobin group 1, 3, 5 and 7 days after infarction. The volume of the region gradually decreased from 1 day to 7 days after infarction. The ratio of NAA/Cho + Cr in the region of T2 hyperintensity in the infarction group was significantly lower than that in the corresponding region in the sham-operated group 3, 5 and 7 days after infarction respectively (P < 0.05). Lac appeared in the region of T2 hyperintensity in the infarction group 1, 3, 5 and 7 days after infarction, but it was not observed in the corresponding region in sham-operated group at all time points. Compared with the sham-operated group, the ratios of beta ATP/PME + PDE and PCr/PME + PDE of the whole brain in the infarction group were significantly lower 1, 3 and 5 days after infarction respectively (P < 0.05), and the ratio of beta ATP/PCr also was significantly lower 1 day after infarction (P < 0.05). Batroxobin significantly decreased the volume of the region of T2 hyperintensity 1 and 3 days after infarction (P < 0.05), significantly increased the ratio of NAA/Cho + Cr in the region 5 and 7 days after infarction (P < 0.05), significantly decreased the ratios of Lac/Cho + Cr and Lac/NAA in the region 5 and 7 days after infarction (P < 0.05), and significantly increased the ratios of beta ATP/PME + PDE and beta ATP/PCr in the whole brain 1 day after infarction (P < 0.05). The results indicated that the infracted region had severe edema, increased Lac and apparent neuronal dysfunction and death, and energy metabolism of the whole brain decreased after focal infarction, and that batroxobin effectively ameliorated the above-mentioned abnormal changes.

Animals↗
PubMed2001

[Effect of Rheum palmatum decoction on increasing intelligence].

OBJECTIVE: To observe the nootropic effect of Rheum palmatum decoction in mice. METHOD: Step-down test. RESULTS: Rheum palmatum decoction could increase learning and memory ability of old mice and also improve the impairments of memory acquisition, memory consolidation and memory retrieval induced by scopalamine, NaNO2 and 40% EtOH respectively in mice. CONCLUSION: Rheum palmatum decoction contains the substances of increasing intelligence which may be substantial basis of the decoctions mainly containing Rheum palmatum theraping aphasia from apoplexy, senile dysmnesis, sequel from brain trauma, etc.

Animals↗
PubMed2001

[Rigid registration and fusion between SPECT and MRI/CT based on external fiducial alignment].

A convenient and effective method of rigid registration and fusion between SPECT and MRI/CT which is based on external fiducial alignment system is designed. After thresholding the image, many image processing arithmetics such as erosion, dilation are used to ensure the corresponding points which will be used to registrate. Then these points are used to minimize the cost function. In this way, an optimal rigid transform is gotten to registrate the images accurately and fuse them well. Error in the registration and fusion is reduced furthest by well designing the N-shape bracket which is used as the external fiducial alignment system and also by limiting the relative displacement between the bracket and the patient. Experimentation shows that this method can realize the registration and fusion of SPECT and MRI/CT effectively.

Algorithms↗
PubMed2000

Control of viremia and prevention of clinical AIDS in rhesus monkeys by cytokine-augmented DNA vaccination.

With accumulating evidence indicating the importance of cytotoxic T lymphocytes (CTLs) in containing human immunodeficiency virus-1 (HIV-1) replication in infected individuals, strategies are being pursued to elicit virus-specific CTLs with prototype HIV-1 vaccines. Here, we report the protective efficacy of vaccine-elicited immune responses against a pathogenic SHIV-89.6P challenge in rhesus monkeys. Immune responses were elicited by DNA vaccines expressing SIVmac239 Gag and HIV-1 89.6P Env, augmented by the administration of the purified fusion protein IL-2/Ig, consisting of interleukin-2 (IL-2) and the Fc portion of immunoglobulin G (IgG), or a plasmid encoding IL-2/Ig. After SHIV-89.6P infection, sham-vaccinated monkeys developed weak CTL responses, rapid loss of CD4+ T cells, no virus-specific CD4+ T cell responses, high setpoint viral loads, significant clinical disease progression, and death in half of the animals by day 140 after challenge. In contrast, all monkeys that received the DNA vaccines augmented with IL-2/Ig were infected, but demonstrated potent secondary CTL responses, stable CD4+ T cell counts, preserved virus-specific CD4+ T cell responses, low to undetectable setpoint viral loads, and no evidence of clinical disease or mortality by day 140 after challenge.

AIDS Vaccines↗
PubMed2000

An islet-specific CD8+ T cell hybridoma generated from non-obese diabetic mice recognizes insulin as an autoantigen.

Although CD8+ T cells play a major role in beta cell destruction in insulin-dependent diabetes in the non-obese diabetic mouse, the T cell autoantigen(s) recognized by such cells remains to be identified. Therefore, an islet-reactive, CD8+ T cell line was generated from islet-infiltrating cells and hybridized by fusion with a CD8+ alphabeta TCR- BW5147 thymoma. In the presence of islets, none of the 12 CD3+ CD8+ T cell hybridomas isolated secreted IL-2/IL-4 or IFNgamma but three were islet specific, as shown by activation induced cell death. Subclone 4A7.7.15 recognized only islets expressing H-2Kd, demonstrated islet-specific inhibition of proliferation and concomitant partial arrest in the G2/M phase of the cell cycle. Further analysis using a panel of cell lines, expressing H-2Kd, and transfected with the cDNA for various putative autoantigens in type 1 diabetes showed that 4A7.7.15 recognizes insulin as an antigen.

Animals↗
PubMed2000

A CAD system for the automatic detection of clustered microcalcifications in digitized mammogram films.

Clusters of microcalcifications in mammograms are an important early sign of breast cancer. This paper presents a computer-aided diagnosis (CAD) system for the automatic detection of clustered microcalcifications in digitized mammograms. The proposed system consists of two main steps. First, potential microcalcification pixels in the mammograms are segmented out by using mixed features consisting of wavelet features and gray level statistical features, and labeled into potential individual microcalcification objects by their spatial connectivity. Second, individual microcalcifications are detected by using a set of 31 features extracted from the potential individual microcalcification objects. The discriminatory power of these features is analyzed using general regression neural networks via sequential forward and sequential backward selection methods. The classifiers used in these two steps are both multilayer feedforward neural networks. The method is applied to a database of 40 mammograms (Nijmegen database) containing 105 clusters of microcalcifications. A free-response operating characteristics (FROC) curve is used to evaluate the performance. Results show that the proposed system gives quite satisfactory detection performance. In particular, a 90% mean true positive detection rate is achieved at the cost of 0.5 false positive per image when mixed features are used in the first step and 15 features selected by the sequential backward selection method are used in the second step. However, we must be cautious when interpreting the results, since the 20 training samples are also used in the testing step.

Breast Neoplasms↗
PubMed2000

The relationship between angiotensinogen gene CD235 met-->Thr substitution polymorphism and brain infarction in Chinese.

OBJECTIVE: To detect the relationship between angiotensinogen (AGT) gene CD235 Met-->Thr substitution polymorphism and brain infarction. METHODS: The CD235Met-->Thr substitution polymorphism in exon 2 of AGT gene was analyzed in normal group(82 cases) and brain infarction group (102 cases) by a combination of polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP). RESULTS: Between the control group and brain infarction group, chi-square test showed no significant difference in the frequencies of T/T genotype and T allele (P>0.05). However, the frequencies of T/T genotype and T allele (70.6% and 83.8%) for multiple brain infarction patients were higher than those (42.6% and 65.2%) for lacunae brain infarction patients and those (42.0% and 65.2%) for controls (P<0.01). The frequencies of AGT T/T genotype and T allele (65.3% and 78.6%) for patients with brain infarction associated with hypertension were higher than those (42.0% and 65.0%) for patients with brain infarction not associated with hypertension (P<0.05). CONCLUSION: No direct relationship between AGT CD235 Met-->Thr substitution polymorphism and ordinary brain infarction has been observed, but there is a significant correlation between AGT gene T/T genotype and multiple brain infarction, and brain infarction associated with hypertension too.

Adult↗
PubMed2000

[The development of a new version of multifunctional radioactive nuclide diagnostic instrument].

A new kind of multifunctional radioactive nuclide diagnostic instrument is described here, which can make noninvasive measurements on the metabolic functions of five most important inner organs which are kidney, liver, heart and thyroid. The design of the hardware and software of the new instrument embodies many advanced advantages such as multifunctional integration, high accuracy and high reliability, computer controlled, automatic data-processing, easy to use and manage, and so on. Applications show that this is an advanced and efficient radioactive medical diagnostic instrument.

Equipment Design↗
PubMed1999

Propagating activation during oscillations and evoked responses in neocortical slices.

Population activity in the cortex is poorly understood. In this report we use voltage-sensitive dye imaging to examine the spatiotemporal patterns of a 7-10 Hz oscillation in neocortical slices from rat somatosensory areas. This oscillation appeared as a component of spontaneous epochs when the preparation was bathed in low [Mg] artificial CSF (ACSF) (Silva et al., 1991). Each epoch started with a synchronized spike, and 3-200 cycles of oscillation emerged afterward. Voltage-sensitive dye imaging revealed that the oscillations in the local field potential recordings were actually caused by a propagating population activation. This activation propagated in a relatively uniform size (not expanding). We call this confined, propagating activation a "dynamic ensemble." During each oscillation cycle, one (occasionally two) dynamic ensemble(s) appeared in the slice and was sustained for 60-200 msec. Dynamic ensembles propagated at approximately 30 mm/sec; the activity could propagate in both directions in cortical slices. The propagation consisted in part of "jumps," the locations of which were not fixed. Dynamic ensembles were distinguishable from the epileptiform spikes that occurred in low [Mg] ACSF. Population events similar to dynamic ensembles were also evoked under conditions of unaltered excitability (slice in normal ACSF) by electrical stimulation that activated a low density of neurons in a large area. Our data suggest that self-sustained, spatially confined, and propagating dynamic ensembles might be related to the epoch oscillations in somatosensory cortex seen in vivo (Nicolelis et al., 1995) and thus resemble one form of population activation in the neocortex.

Animals↗
PubMed1999

Membrane topology of the xenobiotic-exporting subunit, MexB, of the MexA,B-OprM extrusion pump in Pseudomonas aeruginosa.

The MexA,B-OprM efflux pump assembly of Pseudomonas aeruginosa consists of two inner membrane proteins and one outer membrane protein. The cytoplasmic membrane protein, MexB, appears to function as the xenobiotic-exporting subunit, whereas the MexA and OprM proteins are supposed to function as the membrane fusion protein and the outer membrane channel protein, respectively. Computer-aided hydropathy analyses of MexB predicted the presence of up to 17 potential transmembrane segments. To verify the prediction, we analyzed the membrane topology of MexB using the alkaline phosphatase gene fusion method. We obtained the following unique characteristics. MexB bears 12 membrane spanning segments leaving both the amino and carboxyl termini in the cytoplasmic side of the inner membrane. Both the first and fourth periplasmic loops had very long hydrophilic domains containing 311 and 314 amino acid residues, respectively. This fact suggests that these loops may interact with other pump subunits, such as the membrane fusion protein MexA and the outer membrane protein OprM. Alignment of the amino- and the carboxyl-terminal halves of MexB showed a 30% homology and transmembrane segments 1, 2, 3, 4, 5, and 6 could be overlaid with the segments 7, 8, 9, 10, 11, and 12, respectively. This result suggested that the MexB has a 2-fold repeat that strengthen the experimentally determined topology model. This paper reports the structure of the pump subunit, MexB, of the MexA,B-OprM efflux pump assembly. This is the first time to verify the topology of the resistant-nodulation-division efflux pump protein.

Amino Acid Sequence↗