Biomedical subjects

Hong Cao

Publications and source records attributed to Hong Cao.

At least 55 records · Page 3Linked to original sources

PubMed2003

The large GTPase dynamin is required for hepatitis B virus protein secretion from hepatocytes.

BACKGROUND/AIMS: The hepatocellular transport pathways and cellular proteins utilized during the packaging and secretion of hepatitis B virus are poorly understood. In this study, we tested if the large GTPase dynamin, a protein involved in vesicle formation and secretion at the trans-Golgi network in hepatocytes, is also used by hepatitis B virus (HBV) in secreting viral proteins. METHODS: Using HepG2.2.15 cells expressing the full-length HBV genome, we tested the effects of wild-type and mutant dynamin on the localization and secretion of two hepatitis B antigens, hepatitis B surface antigen (HBsAg) and hepatitis B e antigen (HBeAg). Distribution of these two antigens was analyzed morphologically in cells transiently transfected with wild-type or mutant dynamin constructs, whereas secretion of the antigens was measured by testing for antigen levels in the media of transfected cells. RESULTS: Mutant dynamin was found to induce a striking redistribution of HBsAg and HBeAg to a perinuclear compartment, as well as a decrease in the levels of HBsAg and HBeAg present in cell culture media indicating a reduction in viral protein secretion. At the electron microscopy level, cells expressing the mutant dynamin showed a marked accumulation of viral particles in dilated cisternae of an uncharacterized cellular compartment. CONCLUSIONS: Intact dynamin function is required for secretion of HBV proteins from hepatocytes through an uncharacterized cellular compartment.

Biological Transport↗

PubMed2003

Regulation of anti-phosphatidylserine antibodies.

The degree of heavy chain (H) editing, the types of Vkappa editors, and the pattern of Jkappa usage are correlated with a range of the affinity of anti-DNA. This range was determined by the number and location of arginine (R) residues in the VH. We, here, changed a key arginine residue in the VH of anti-DNA transgene to glycine, which sharply reduces the affinity for dsDNA. However, complete reversion of this anti-DNA to germline enhances the affinity for phosphatidylserine (PS). The B cells of this low-affinity anti-DNA and anti-PS transgenic mouse are tightly regulated by receptor editing. Thus, anti-PS B cells are another example of a constitutive self-antigen regulated in the bone marrow.

Animals↗

PubMed2003

A dynamin-cortactin-Arp2/3 complex mediates actin reorganization in growth factor-stimulated cells.

The mechanisms by which mammalian cells remodel the actin cytoskeleton in response to motogenic stimuli are complex and a topic of intense study. Dynamin 2 (Dyn2) is a large GTPase that interacts directly with several actin binding proteins, including cortactin. In this study, we demonstrate that Dyn2 and cortactin function to mediate dynamic remodeling of the actin cytoskeleton in response to stimulation with the motogenic growth factor platelet-derived growth factor. On stimulation, Dyn2 and cortactin coassemble into large, circular structures on the dorsal cell surface. These "waves" promote an active reorganization of actin filaments in the anterior cytoplasm and function to disassemble actin stress fibers. Importantly, inhibition of Dyn2 and cortactin function potently blocked the formation of waves and subsequent actin reorganization. These findings demonstrate that cortactin and Dyn2 function together in a supramolecular complex that assembles in response to growth factor stimulation and mediates the remodeling of actin to facilitate lamellipodial protrusion at the leading edge of migrating cells.

Actin-Related Protein 3↗

PubMed2003

Noncontact radio-frequency ablation for obtaining deeper lesions.

Radio-frequency (RF) cardiac catheter ablation has been very successful for treating some cardiac arrhythmias, however, the success rate for ventricular tachycardias is still not satisfactory. Some existing methods for developing deeper lesions include active cooling of the electrode and modifying the electrode shape. We propose a method of noncontact ablation, to solve this problem. We apply 120 W of power through an 8-mm electrode for a 120-s duration, with distances from 0 to 3 mm between electrode and myocardium, to create lesions in myocardium. We apply flow rates of 1, 3, and 5 L/min to determine their effect. Results show that with an optimal distance from 0.5 to 1.5 mm between electrode and myocardium, we increase lesion depth from 7.5 mm for contact ablation to 9.5 mm for noncontact ablation. For different flow rates, the optimal distance various. The effect of flow rate is not obvious. Higher flow rate does not lead to a deeper lesion.

Animals↗

PubMed2003

In vitro measurement of myocardial impedivity anisotropy with a miniature rectangular tube.

Due to rapid change of fiber orientation, it is difficult to measure myocardial impedivity separately in a longitudinal or transverse fiber direction without mutual influence in the two directions. Previously published values of the longitudinal and the transverse myocardial impedivity were derived indirectly from measurements that mixed the impedivity in all directions. Those values are questionable because the derivations were based on a simplified uniform myocardial fiber model. In this paper, a miniature rectangular tube was devised to facilitate direct measurement of myocardial impedivity in a uniform fiber direction. The average transverse-to-longitudinal ratio of the measured in vitro swine myocardial impedivity was about 1.66 from 1 Hz to 1 kHz and dropped to 1.25 at 1 MHz. The result is important for accurate modeling of the electrical property of myocardium in biomedical research of radio-frequency cardiac catheter ablation.

Animals↗

PubMed2003

Cortactin is a component of clathrin-coated pits and participates in receptor-mediated endocytosis.

The actin cytoskeleton is believed to contribute to the formation of clathrin-coated pits, although the specific components that connect actin filaments with the endocytic machinery are unclear. Cortactin is an F-actin-associated protein, localizes within membrane ruffles in cultured cells, and is a direct binding partner of the large GTPase dynamin. This direct interaction with a component of the endocytic machinery suggests that cortactin may participate in one or several endocytic processes. Therefore, the goal of this study was to test whether cortactin associates with clathrin-coated pits and participates in receptor-mediated endocytosis. Morphological experiments with either anti-cortactin antibodies or expressed red fluorescence protein-tagged cortactin revealed a striking colocalization of cortactin and clathrin puncta at the ventral plasma membrane. Consistent with these observations, cells microinjected with these antibodies exhibited a marked decrease in the uptake of labeled transferrin and low-density lipoprotein while internalization of the fluid marker dextran was unchanged. Cells expressing the cortactin Src homology three domain also exhibited markedly reduced endocytosis. These findings suggest that cortactin is an important component of the receptor-mediated endocytic machinery, where, together with actin and dynamin, it regulates the scission of clathrin pits from the plasma membrane. Thus, cortactin provides a direct link between the dynamic actin cytoskeleton and the membrane pinchase dynamin that supports vesicle formation during receptor-mediated endocytosis.

Actins↗

PubMed2003

[Determination of aloin in aloes by HPLC].

OBJECTIVE: To describe a simple and rapid isocratic reversed-phase high performance liquid chromatography method for the baseline separation, identification and assay of aloin in aloes. METHOD: The analytical column was a ZORBAX SB-C18(4.6 mm x 250 mm) filled with a 5 microns stationary phase. The mobile phase consisted of acetonitrile-water (25:75); the flow-rate was 1 mL.min-1. The injection volume was 10 microL. The DAD detector was set at 355 nm. RESULT: The calibration curve was linear over the range of 0.17-5.9 micrograms (r = 0.9999). The average recovery of the method was 98.6%, RSD 1.32% (n = 6). CONCLUSION: The results showed that this method was reliable and accurate. The method was applied to eleven Cape and East African aloes of different origin.

Aloe↗

PubMed2002

Specific HIV-1 TAR RNA loop sequence and functional groups are required for human cyclin T1-Tat-TAR ternary complex formation.

Replication of human immunodeficiency virus requires Tat protein which activates elongation of RNA polymerase II transcription at the HIV-1 promoter through interaction with the cyclin T1 (CycT1) subunit of the positive transcription elongation factor complex (P-TEFb). Tat binds directly through its transactivation domain to the CycT1 subunit of the P-TEFb and induces loop sequence specific binding of the P-TEFb onto nascent HIV-1 TAR RNA. By using a gel electrophoresis method and a comprehensive set of TAR loop mutants, we have identified the sequence and structural determinants for high-affinity CycT1-Tat-TAR ternary complex formation. Our results show that CycT1 and Tat binding to TAR RNA is highly cooperative, and a capacity of 85%, a Hill coefficient of 2.7, and a dissociation constant (K(D)) of 2.45 nM were observed. These results indicate that there are three binding sites on TAR RNA. CycT1 does not bind TAR RNA in the absence of Tat, and Tat binding to TAR, while detectable, is very inefficient in the absence of CycT1. It is conceivable that the CycT1-Tat heterodimer directly binds to TAR RNA in the U-rich RNA bulge region and this binding facilitates the interactions of the CycT1-Tat heterodimer at the other two sites in the RNA loop region. On the basis of our results, we suggest a model where CycT1 interacts with Tat protein and positions the protein complex to make contacts with the G34 region of the loop sequence; G34 is critical for CycT1-Tat binding and forms a C30.G34 base pair. Two functional groups, O6 and N7, at nucleotide positions 32 and 34 in the TAR loop are essential for CycT1-Tat interactions with TAR RNA. The identity of two nucleotides, U31 and G33, is not critical, but they contribute to the stabilization of the RNA-protein complex. The presence of a single-nucleotide bulge of A35 or C35 is essential for distortion of the backbone RNA structure as well as the accessibility of functional groups in the major groove of the double-helical region. CycT1-Tat interaction with TAR RNA represents another example of the flexibility and complexity of RNA structure involved in protein recognition.

Base Sequence↗

PubMed2002

[Age related changes of serum sex hormone levels and tartrate-resistant acid phosphatase activity in healthy women].

OBJECTIVE: To study the changes of serum sex hormone levels and tartrate-resistant acid phosphatase (TRACP) activity in healthy women of different ages, and to provide parameters for basic and clinical research of osteoporosis. METHODS: Serum sex hormone (FSH, LH, E2, P) levels and TRACP activity were measured in 236 premenopausal healthy women aged 23-53 years and divided into 3 groups, and in 91 postmenopausal healthy women aged 48-71 years. RESULTS: Except serum FSH levels which were significantly higher in the third group than those of the other groups in premenopausal healthy women (P < 0.05), no significant difference in sex hormone levels and TRACP activity was found among the 3 groups; the postmenopausal group showed a significant increase in serum FSH, LH levels and TRACP activity and a significant decrease in serum E2, P levels, compared with the premenopausal groups (P < 0.01). There was a negative correlation between E2, P levels and TRACP activity in the postmenopausal women (r = -0.41 and -0.37, respectively, P < 0.01). CONCLUSION: Postmenopausal diminution of E2, P results in an increase of serum FSH and LH levels, and may bring about an increase of bone resorption, which results in the increase of TRACP activity.

Acid Phosphatase↗

PubMed2002

Quinolinate phosphoribosyltransferase: kinetic mechanism for a type II PRTase.

Quinolinate phosphoribosyltransferase (QAPRTase, EC 2.4.2.19) catalyzes the formation of nicotinate mononucleotide, carbon dioxide, and pyrophosphate from 5-phosphoribosyl 1-pyrophosphate (PRPP) and quinolinic acid (QA, pyridine 2,3-dicarboxylic acid). The enzyme is the only type II PRTase whose X-ray structure is known. Here we determined the kinetic mechanism of the enzyme from Salmonella typhimurium. Equilibrium binding studies show that PRPP and QA each form binary complexes with the enzyme, with K(D) values (53 and 21 microM, respectively) similar to their K(M) values (30 and 25 microM, respectively). Although neither PP(i) nor NAMN products bound well to the enzyme, 130-fold tighter binding of PP(i) (K(D) = 75 microM) and NAMN (K(D) = 6 microM) in a ternary complex was observed. Phthalic acid (K(D) = 21 microM) and PRPP each caused a 2.5-fold tightening of the other's binding. Isotope trapping experiments indicated that the E.QA complex is catalytically competent, whereas the E.PRPP complex could not be trapped. Pre-steady-state kinetics gave a linear rate of NAMN formation, indicating that on-enzyme phosphoribosyl transfer chemistry is rate-determining. Isotope trapping from the steady state revealed that nearly all QA and about one-third of PRPP in ternary enzyme.QA.PRPP complexes could be trapped as the product. Substrate inhibition by PRPP was observed. These data demonstrate a predominantly ordered kinetic mechanism in which productive binding of quinolinic acid precedes that of PRPP. An E.PRPP complex exists as a nonproductive side branch.

Isotopes↗

PubMed2002

Mechanical compliance of the endocardium.

Radio-frequency (RF) ablation is an accepted treatment for cardiac arrhythmias related to abnormal focal cardiac substrate. The penetration depth of the electrode into the endocardium affects lesion size, a critical determinant of success of RF ablation. We measured the relation between the mechanical compliance and the penetration depth of RF ablation catheter electrode at frequently ablated areas of the endocardium and examined the influence of time after death on mechanical properties of the tissue. We measured force versus time for eight insertion depths of the catheter electrode into full-thickness endocardial samples derived from the mitral valve annulus, the left ventricular free wall and the tricuspid valve annulus. We varied the time after death at 15, 40 min, 3, 8, and 18 h and repeated our measurements. At 15 min after death, the first 0.5mm penetration depth caused the fastest relaxation at 55 s. Force decay decreased dramatically at 15 min after death as the penetration depth increased from 0.5 to 4mm. We used the force data sampled at 60s after insertion to approximate the elasticity. We observed the relations between the force versus the insertion depth. The force increased by a factor of 5 for the mitral valve annulus and 8 for the left free wall from 15 min to 18 h. We derived coefficients of a second-order polynomial equation relating the force data to insertion depth with R(2)>0.99.

Animals↗

PubMed2002

Oxidative stress-induced calcium signalling in Aspergillus nidulans.

The effects of oxidative stress on levels of calcium ion (Ca(2+)) in Aspergillus nidulans were measured using strains expressing aequorin in the cytoplasm (Aeq(cyt)) and mitochondria (Aeq(mt)). When oxidative stress was induced by exposure to 10-mM H(2)O(2), the mitochondrial calcium response (Ca(mt)(2+)) was greater than the change in cytoplasmic calcium (Ca(c)(2+)). The Ca(mt)(2+) response to H(2)O(2) was dose dependent, while the increase in [Ca(c)(2+)] did not change with increasing H(2)O(2). The increase in both [Ca(c)(2+)] and [Ca(mt)(2+)] in response to oxidative stress was enhanced by exposure of cells to Ca(2+). The presence of chelator in the external medium only partially inhibited the Ca(mt)(2+) and Ca(c)(2+) responses to oxidative stress. Reagents that alter calcium fluxes had varied effects on the Ca(mt)(2+) response to peroxide. Ruthenium red blocked the increase in [Ca(mt)(2+)], while neomycin caused an even greater increase in [Ca(mt)(2+)]. Treatment with ruthenium red and neomycin had no effect on the Ca(c)(2+) response. Bafilomycin A and oligomycin had no effect on either the mitochondrial or cytoplasmic response. Inhibitors of both voltage-regulated calcium channels and intracellular calcium release channels inhibited the Ca(2+)-dependent component of the Ca(mt)(2+) response to oxidative stress. We conclude that the more significant Ca(2+) response to oxidative stress occurs in the mitochondria and that both intracellular and extracellular calcium pools can contribute to the increases in [Ca(c)(2+)] and [Ca(mt)(2+)] induced by oxidative stress.

Aequorin↗

PubMed2002

Modeling bipolar phase-shifted multielectrode catheter ablation.

Atrial fibrillation (AFIB) is a common clinical problem affecting approximately 0.5-1% of the United States population. Radio-frequency (RF) multielectrode catheter (MEC) ablation has successes in curing AFIB. We utilized finite-element method analysis to determine the myocardial temperature distribution after 30 s, 80 degrees C temperature-controlled unipolar ablation using three 7F 12.5-mm electrodes with 2-mm interelectrode spacing MEC. Numerical results demonstrated that cold spots occurred at the edges of the middle electrode and hot spots at the side electrodes. We introduced the bipolar phase-shifted technique for RF energy delivery of MEC ablation. We determined the optimal phase-shift (phi) between the two sinusoidal voltage sources of a simplified two-dimensional finite-element model. At the optimal phi, we can achieve a temperature distribution that minimizes the difference between temperatures at electrode edges. We also studied the effects of myocardial electric conductivity (sigma), thermal conductivity (k), and the electrode spacing on the optimal phi. When we varied sigma and kappa from 50% to 150%, optimal phi ranged from 29.5 degrees to 23.5 degrees, and in the vicinity of 26.5 degrees, respectively. The optimal phi for 3-mm spacing MEC was 30.5 degrees. We show the design of a simplified bipolar phase-shifted MEC ablation system.

Atrial Fibrillation↗

PubMed2002

Using electrical impedance to predict catheter-endocardial contact during RF cardiac ablation.

During radio-frequency (RF) cardiac catheter ablation, there is little information to estimate the contact between the catheter tip electrode and endocardium because only the metal electrode shows up under fluoroscopy. We present a method that utilizes the electrical impedance between the catheter electrode and the dispersive electrode to predict the catheter tip electrode insertion depth into the endocardium. Since the resistivity of blood differs from the resistivity of the endocardium, the impedance increases as the catheter tip lodges deeper in the endocardium. In vitro measurements yielded the impedance-depth relations at 1, 10, 100, and 500 kHz. We predict the depth by spline curve interpolation using the obtained calibration curve. This impedance method gives reasonably accurate predicted depth. We also evaluated alternative methods, such as impedance difference and impedance ratio.

Animals↗

PubMed2002

In-vivo measurement of swine myocardial resistivity.

We used a four-terminal plunge probe to measure myocardial resistivity in two directions at three sites from the epicardial surface of eight open-chest pigs in-vivo at eight frequencies ranging from 1 Hz to 1 MHz. We calibrated the plunge probe to minimize the error due to stray capacitance between the measured subject and ground. We calibrated the probe in saline solutions contained in a metal cup situated near the heart that had an electrical connection to the pig's heart. The mean of the measured myocardial resistivity was 319 ohm x cm at 1 Hz down to 166 ohm x cm at 1 MHz. Statistical analysis showed the measured myocardial resistivity of two out of eight pigs was significantly different from that of other pigs. The myocardial resistivity measured with the resistivity probe oriented along and across the epicardial fiber direction was significantly different at only one out of the eight frequencies. There was no significant difference in the myocardial resistivity measured at different sites.

Animals↗

PubMed2002

Error analysis of tissue resistivity measurement.

We identified the error sources in a system for measuring tissue resistivity at eight frequencies from 1 Hz to 1 MHz using the four-terminal method. We expressed the measured resistivity with an analytical formula containing all error terms. We conducted practical error measurements with in-vivo and bench-top experiments. We averaged errors at all frequencies for all measurements. The standard deviations of error of the quantization error of the 8-bit digital oscilloscope with voltage averaging, the nonideality of the circuit, the in-vivo motion artifact and electrical interference combined to yield an error of +/- 1.19%. The dimension error in measuring the syringe tube for measuring the reference saline resistivity added +/- 1.32% error. The estimation of the working probe constant by interpolating a set of probe constants measured in reference saline solutions added +/- 0.48% error. The difference in the current magnitudes used during the probe calibration and that during the tissue resistivity measurement caused +/- 0.14% error. Variation of the electrode spacing, alignment, and electrode surface property due to the insertion of electrodes into the tissue caused +/- 0.61% error. We combined the above errors to yield an overall standard deviation error of the measured tissue resistivity of +/- 1.96%.

Animals↗

PubMed2002

FEM analysis of predicting electrode-myocardium contact from RF cardiac catheter ablation system impedance.

We used the finite-element method (FEM) to model and analyze the resistance between the catheter tip electrode and the dispersive electrode during radio-frequency cardiac catheter ablation for the prediction of myocardium-electrode contact. We included deformation of the myocardial surface to achieve accurate modeling. For perpendicular catheter contact, we measured the side view of myocardial deformation using X-ray projection imaging. We averaged the deformation contour from nine samples, and then incorporated the contour information into our FEM model. We measured the resistivity of the bovine myocardium using the four-electrode method, and then calculated the resistance change as the catheter penetrated into the myocardium. The FEM result of resistance versus catheter penetration depth matches well with our experimental data.

Animals↗

PubMed2002

Application of nanotechnology in biomedical sciences.

Nanotechnology, a new research field that holds enormous prospects in the 21th century, has by now gained wide application in biomedical sciences, and consequently gives rise to two new cross-disciplines, nanobiology and nanobiomedicine. The authors provide a brief summarization of the progress so far achieved in these two new disciplines.

Biomedical Research↗