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Biomedical subjects

J Xiong

Publications and source records attributed to J Xiong.

At least 73 records · Page 4Linked to original sources

Construction and characterization of a photosystem II D1 mutant (arginine-269-glycine) of Chlamydomonas reinhardtii.

Numerous lines of evidence indicate that bicarbonate anion regulates electron and proton transfer processes in the photosystem II (PSII) complex of chloroplasts and cyanobacteria. On the reducing side of PSII, the addition of bicarbonate to bicarbonate-depleted (or formate-treated) membranes accelerates, especially, QA(-)-->QB(-) electron transfer kinetics. The site(s) at which bicarbonate binds is unknown. It is evident, however, from several spectroscopic studies that the bicarbonate binding site on the reducing side of PSII includes the non-heme iron located between the QA and QB sites. Since small anions may displace bicarbonate (Good, N.E. (1963) Plant Physiol. 38, 298-304) [1], it is apparent that the bicarbonate binding site is electrostatic in nature, presumably also involving positively charged amino acid residues. Previously, it had been predicted that residue arginine 269 of the PSII D1 protein may participate in bicarbonate binding. To test this hypothesis, we have generated a non-conservative mutation in the psbA gene of Chlamydomonas reinhardtii which converts residue R269 to a glycine (R269G). The R269G mutant was unable to grow photosynthetically or evolve oxygen. This phenotype is associated with a lack of the tetra-manganese water splitting complex and a reduced capacity to form a stabilized charge separated state (defined as TyrD+/QA- under the experimental conditions measured). In addition, the mutant cells have a less stable PSII complex than wild-type cells, particularly when grown in the light. It is apparent from analyses of the effect of formate on the magnitude of the QA-Fe+2 EPR signal, however, that the bicarbonate or formate binding site is not substantially affected by the R269G mutation. Although our results do not substantiate that residue R269 is the site at which bicarbonate is bound, they demonstrate the importance of R269 in the structure and function of PSII. It is apparent from analysis of the photosynthetic phenotype, that the structural perturbations on the stromal side of the D1 protein are transduced to the lumenal side of the membrane altering charge accumulating processes on the electron donor side of PSII.

Animals↗

TRAF6 is a signal transducer for interleukin-1.

Many cytokines signal through different cell-surface receptors to activate the transcription factor NF-kappaB. Members of the TRAF protein family have been implicated in the activation of NF-kappaB by the tumour-necrosis factor (TNF)-receptor superfamily. Here we report the identification of a new TRAF family member, designated TRAF6. When overexpressed in human 293 cells, TRAF6 activates NF-kappaB. A dominant-negative mutant of TRAF6 inhibits NF-kappaB activation signalled by interleukin-1 (IL-1) but not by TNF. IL-1 treatment of 293 cells induces the association of TRAF6 with IRAK, a serine/threonine kinase that is rapidly recruited to the IL-1 receptor after IL-1 induction. These findings indicate that TRAF proteins may function as signal transducers for distinct receptor families and that TRAF6 participates in IL-1 signalling.

Amino Acid Sequence↗

I-TRAF is a novel TRAF-interacting protein that regulates TRAF-mediated signal transduction.

Tumor necrosis factor (TNF) receptor-associated factor (TRAF) proteins associate with and transduce signals from TNF receptor 2, CD40, and presumably other members of the TNF receptor superfamily. TRAF2 is required for CD40- and TNF-mediated activation of the transcription factor NF-kappa B. Here we describe the isolation and characterization of a novel TRAF-interacting protein, I-TRAF, that binds to the conserved TRAF-C domain of the three known TRAFs. Overexpression of I-TRAF inhibits TRAF2-mediated NF-kappa B activation signaled by CD40 and both TNF receptors. Thus, I-TRAF appears as a natural regulator of TRAF function that may act by maintaining TRAFs in a latent state.

Adaptor Proteins, Signal Transducing↗

Cerebellum implicated in sensory acquisition and discrimination rather than motor control.

Recent evidence that the cerebellum is involved in perception and cognition challenges the prevailing view that its primary function is fine motor control. A new alternative hypothesis is that the lateral cerebellum is not activated by the control of movement per se, but is strongly engaged during the acquisition and discrimination of sensory information. Magnetic resonance imaging of the lateral cerebellar output (dentate) nucleus during passive and active sensory tasks confirmed this hypothesis. These findings suggest that the lateral cerebellum may be active during motor, perceptual, and cognitive performances specifically because of the requirement to process sensory data.

Adult↗

Improving the temporal resolution of functional MR imaging using keyhole techniques.

Using a keyhole technique, it is shown that the data acquisition rate of gradient-echo imaging for functional MRI (fMRI) studies can be increased substantially. The resulting enhancement of the temporal resolution of fMRIs was accomplished without modifying the hardware of a conventional MRI system. High spatial resolution fMRI images were first collected with conventional full k-space acquisition and image reconstruction. Using the same data set, simulation reconstruction using the keyhole principle and zero-padding were performed for comparison with the full k-space reconstruction. No significant changes were found for fMRI images generated from the keyhole technique with a data sharing profile of 50% of the k-space. As k-space data sharing profiles increased to 75 and 87.5%, the keyhole fMRI images began to show only modest changes in activation intensity and area compared with the standard images. In contrast, zero-padding fMRI images produced a significant disparity both in activation intensity and area relative to the truly high-resolution fMRI images. The keyhole technique's ability to retain the intensity and area of fMRI information, while substantially reducing acquisition time, makes it a promising method for fMRI studies.

Adult↗

Quantitative assessment of blood inflow effects in functional MRI signals.

Functional MRI (fMRI) signal dependence on changes in blood flow velocities were analyzed for both conventional and echo-planar (EPI) gradient-echo pulse sequences. As the flow velocity increases, the fMRI signal increases monotonically in spoiled gradient-echo sequences, while the fMRI signal may increase or decrease in conventional refocused gradient-echo sequences. A larger flip angle generates a larger inflow contribution to the fMRI signal. For conventional gradient-echo sequences, the inflow contribution to the fMRI images is dominated by the cortical draining veins, while its effect on capillaries is generally small and may be negligible in the spoiled sequences. For EPI gradient-echo sequences, the contribution from inflow effects is relatively small, as compared with the blood oxygen level-dependent (BOLD) contribution, to the fMRI signal, not only for capillaries but also for the cortical draining veins.

Blood Flow Velocity↗

Modeling of the D1/D2 proteins and cofactors of the photosystem II reaction center: implications for herbicide and bicarbonate binding.

A three-dimensional model of the photosystem II (PSII) reaction center from the cyanobacterium Synechocystis sp. PCC 6803 was generated based on homology with the anoxygenic purple bacterial photosynthetic reaction centers of Rhodobacter sphaeroides and Rhodopseudomonas viridis, for which the X-ray crystallographic structures are available. The model was constructed with an alignment of D1 and D2 sequences with the L and M subunits of the bacterial reaction center, respectively, and by using as a scaffold the structurally conserved regions (SCRs) from bacterial templates. The structurally variant regions were built using a novel sequence-specific approach of searching for the best-matched protein segments in the Protein Data Bank with the "basic local alignment search tool" (Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ, 1990, J Mol Biol 215:403-410), and imposing the matching conformational preference on the corresponding D1 and D2 regions. The structure thus obtained was refined by energy minimization. The modeled D1 and D2 proteins contain five transmembrane alpha-helices each, with cofactors (4 chlorophylls, 2 pheophytins, 2 plastoquinones, and a non-heme iron) essential for PSII primary photochemistry embedded in them. A beta-carotene, considered important for PSII photoprotection, was also included in the model. Four different possible conformations of the primary electron donor P680 chlorophylls were proposed, one based on the homology with the bacterial template and the other three on existing experimental suggestions in literature. The P680 conformation based on homology was preferred because it has the lowest energy. Redox active tyrosine residues important for P680+ reduction as well as residues important for PSII cofactor binding were analyzed. Residues involved in interprotein interactions in the model were also identified. Herbicide 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) was also modeled in the plastoquinone QB binding niche using the structural information available from a DCMU-binding bacterial reaction center. A bicarbonate anion, known to play a role in PSII, but not in anoxygenic photosynthetic bacteria, was modeled in the non-heme iron site, providing a bidentate ligand to the iron. By modifying the previous hypothesis of Blubaugh and Govindjee (1988, Photosyn Res 19:85-128), we modeled a second bicarbonate and a water molecule in the QB site and we proposed a hypothesis to explain the mechanism of QB protonation mediated by bicarbonate and water. The bicarbonate, stabilized by D1-R257, donates a proton to QB2- through the intermediate of D1-H252; and a water molecule donates another proton to QB2-. Based on the discovery of a "water transport channel" in the bacterial reaction center, an analogous channel for transporting water and bicarbonate is proposed in our PSII model. The putative channel appears to be primarily positively charged near QB and the non-heme iron, in contrast to the polarity distribution in the bacterial water transport channel. The constructed model has been found to be consistent with most existing data.

Amino Acid Sequence↗

Investigation of germination and aging in Moravian III barley grain by nuclear magnetic resonance.

High-resolution, solid-state 1H nuclear magnetic resonance (NMR) techniques are used for the first time to study germination in imbibed Moravian III barley grains. Whereas magic-angle spinning 1H NMR spectra reveal the water and lipid components in barley grains, combined rotation and multiple-pulse spectroscopy techniques provide 1H NMR spectra of grains that reveal the protein and carbohydrate as well as the water and lipid components. Spectra of grains are compared with spectra of model compounds to verify assignments. 1H T1 and T2 measurements using magic-angle spinning only and combined rotation and multiple-pulse spectroscopy techniques provide information about molecular mobility within the grains during inhibition. Some grains were subjected to artificial aging conditions. 1H NMR spectral comparisons are made between normal, viable grains and artificially aged grains.

Biophysical Phenomena↗

Ascaris, people and pigs in a rural community of Jiangxi Province, China.

A longitudinal investigation on natural populations of Ascaris in humans and pigs and an investigation of soil contamination with Ascaris eggs were carried out from June 1993 to June 1994 in 2 villages, Manhu area, Xinjian County, Jiangxi Province, China. Results from these studies indicate that although human ascariasis is endemic there is significant fluctuation in both prevalence and the mean number of eggs/g faeces (epg) of the communities. Fluctuation of age-stratified prevalence and mean epg was detected in children but not in most adult groups. Most cases of human ascariasis were judged to involve low intensities of infection and a typical overdispersion distribution pattern was observed through the year. It was estimated that during the year, nearly half of the eggs discharged in the environment came from infections in children aged between 2 and 15 years which accounted for about 30% of the total population. Soil in and around houses and in vegetable gardens was found to be contaminated by Ascaris eggs and this situation remained relatively stable throughout the year. Monthly developmental rate of Ascaris eggs in soil was detected and the results suggest that the fluctuation in prevalence observed during the year should be directly attributed to the effect of seasonality of egg development. Features of Ascaris infection in pigs were found to be similar to those in humans except for a lower mean intensity of infection. The possibility of cross-infection of Ascaris between human and pig hosts is discussed.

Adolescent↗

Expression of apoptosis-regulatory genes in lung tumour cell lines: relationship to p53 expression and relevance to acquired drug resistance.

As a first step towards elucidating the potential role(s) of bcl-2 and bcl-2-related genes in lung tumorigenesis and therapeutic responsiveness, the expression of these genes has been examined in a panel of lung cancer cell lines derived from untreated and treated patients, and in cell lines selected in vitro for multidrug resistance. Bcl-2 was hyperexpressed in 15 of 16 small-cell lung cancer (SCLC) cell lines and two of five non-small-cell lung cancer (NSCLC) lines compared with normal lung and brain, and hyperexpression was not chemotherapy related. Bcl-x was hyperexpressed in the majority of SCLC and NSCLC cell lines as compared with normal tissues, and all lung tumour lines preferentially expressed bcl-x1-mRNA, the splice variant form that inhibits apoptosis. Bax gene transcripts were hyperexpressed in most SCLC and NSCLC cell lines examined compared with normal adult tissues. Mutant p53 gene expression was detected in the majority of the cell lines and no relationship between p53 gene expression and the expression of either bcl-2, bcl-x or bax was observed. No changes in bcl-2, bcl-x and bax gene expression were observed in multidrug-resistant cell lines compared with their drug-sensitive counterparts.

Adult↗

Rapid core-to-peripheral tissue heat transfer during cutaneous cooling.

Perioperative thermal manipulations are usually directed at the skin surface because methods of directly warming the core are invasive or ineffective. However, inadequate heat flow between peripheral and core compartments will decrease the rate at which core temperature changes. We therefore determined whether core hypothermia is delayed after initiation of surface cooling. Six volunteers were anesthetized with propofol and midazolam, and maintained under three layers of passive insulation for 2.5-4 h. Subsequently, the skin surface was cooled using forced air, 1000 L/min, at 10 degrees C. Isoflurane was added as necessary to maintain arteriovenous shunt vasodilation. Overall heat balance was determined from the difference between cutaneous heat loss (thermal flux transducers) and metabolic heat production (oxygen consumption). Average arm and leg (peripheral) tissue temperatures were determined from 19 intramuscular needle thermocouples, 10 skin temperatures, and "deep" foot temperature. Overall body heat content decreased approximately 234 kcal during 2.5 h of active cooling. Core temperature, which was nearly constant before active cooling, decreased approximately 1.3 degrees C/h. There was no delay between initiation of active cooling and the decrease in core temperature. Furthermore, peripheral (arm and leg) and core (trunk and head) tissue heat contents decreased at virtually the same rates: approximately 50 kcal/h and approximately 47 kcal/h, respectively. These data indicate that there is little restriction of heat flow between peripheral and core tissues in vasodilated, anesthetized subjects.

Adult↗

Isoflurane produces marked and nonlinear decreases in the vasoconstriction and shivering thresholds.

BACKGROUND: Desflurane decreases the vasoconstriction and shivering thresholds disproportionately at high anesthetic concentrations. This result contrasts with the authors' previous report that isoflurane decreases the vasoconstriction threshold linearly. It is surprising that the basic shape of the concentration-response curve should differ with these two otherwise similar anesthetics. Therefore, the hypothesis that isoflurane produces a nonlinear reduction in the vasoconstriction threshold was tested. Because the effect of isoflurane on shivering remains unknown, the extent to which isoflurane reduces the shivering threshold also was determined. METHODS: Eight men volunteered to be studied on four randomly ordered days: (1) a target end-tidal isoflurane concentration of 0.55%, (2) a target concentration of 0.7%, (3) control (no anesthesia) and a target end-tidal concentration of 0.85%, and (4) a target end-tidal concentration of 1.0%. Volunteers were surface-cooled until peripheral vasoconstriction and shivering were observed. We arithmetically compensated for changes in skin temperature using the established linear cutaneous contributions to control for each response. From the calculated thresholds (core temperatures triggering responses at a designated skin temperature of 34 degrees C), the concentration-response relation was determined. RESULTS: Isoflurane administration produced a dose-dependent reduction in the vasoconstriction and shivering thresholds, decreasing each approximately 4.6 degrees C at an end-tidal concentration of 1%. Residual analysis indicated that the vasoconstriction and shivering thresholds were decreased in a nonlinear fashion during isoflurane administration. The vasoconstriction-to-shivering range was 1.5 +/- 0.8 degree C without isoflurane, and did not change significantly during isoflurane administration. CONCLUSIONS: The vasoconstriction-to-shivering range remained unchanged by isoflurane administration. In this regard, the effects of isoflurane are similar to those of desflurane, propofol, and alfentanil. The current data differ from the authors' previous report, in that the dose-dependence for vasoconstriction was nonlinear, with isoflurane reducing the threshold disproportionately at higher anesthetic concentrations. Differing dose-dependence in the two studies may result either because the current study's volunteers were not exposed to surgical stimulation and were given less isoflurane, or because of design limitations in the previous protocol.

Adult↗

Thermoregulatory vasoconstriction does not impede core warming during cutaneous heating.

BACKGROUND: Although forced-air warming rapidly increases intraoperative core temperatures, it is reportedly ineffective postoperatively. A major difference between these two periods is that arteriovenous shunts are usually dilated during surgery, whereas vasoconstriction is uniform in hypothermic postoperative patients. Vasoconstriction may decrease efficacy of warming because its major physiologic purposes are to reduce cutaneous heat transfer and restrict heat transfer between the two thermal compartments. Accordingly, we tested the hypothesis that thermoregulatory vasoconstriction decreases cutaneous transfer of applied heat and restricts peripheral-to-core flow of heat, thereby delaying and reducing the increase in core temperature. METHODS: Eight healthy male volunteers anesthetized with propofol and isoflurane were studied. Volunteers were allowed to cool passively until core temperature reached 33 degrees C. On one randomly assigned day, the isoflurane concentration was reduced, to provoke thermoregulatory arteriovenous shunt vasoconstriction; on the other study day, a sufficient amount of isoflurane was administered to prevent vasoconstriction. On each day, forced-air warming was then applied for 2 h. Peripheral (arm and leg) tissue heat contents were determined from 19 intramuscular needle thermocouples, 10 skin temperatures, and "deep" foot temperature. Core (trunk and head) heat content was determined from core temperature, assuming a uniform compartmental distribution. Time-dependent changes in peripheral and core tissue heat contents were evaluated using linear regression. Differences between the vasoconstriction and vasodilation study days, and between the peripheral and core compartments, were evaluated using two-tailed, paired t tests. Data are presented as means +/-SD; P < 0.01 was considered statistically significant. RESULTS: Cutaneous heat transfer was similar during vasoconstriction and vasodilation. Forced-air warming increased peripheral tissue heat content comparably when the volunteers were vasodilated and vasoconstricted: 48 +/- 7 versus 53 +/- 10 kcal/h. Core compartment tissue heat content increased similarly when the volunteers were vasodilated and vasoconstricted: 51 +/- 8 versus 44 +/- 11 kcal/h. Combining the two study days, the increase in peripheral and core heat contents did not differ significantly: 51 +/- 8 versus 48 +/- 10 kcal/h, respectively. Core temperature increased at essentially the same rate when the volunteers remained vasodilated (1.3 degrees C/h) as when they were vasoconstricted (1.2 degrees C/h). CONCLUSIONS: The authors failed to confirm their hypothesis that thermoregulatory vasoconstriction decreases cutaneous transfer of applied heat and restricts peripheral-to-core flow of heat in anesthetized subjects. The reported difference between intraoperative and postoperative rewarming efficacy may result from nonthermoregulatory anesthetic-induced vasodilation.

Adult↗

The further metabolism of sorbitol-3-phosphate and fructose-3-phosphate in the mature rat lens.

Glucose and phosphorus metabolism in mature (8-month-old) rat lenses were examined with NMR spectroscopy. Nondiabetic mature lenses contained sorbitol-3-phosphate (S3P) and fructose-3-phosphate (F3P) which were absent from young (1- to 2-month-old) normal rat lenses. The concentrations of these two phosphates can be changed through (1) diabetes induction with streptozotocin - this results in a dramatic increase in both compounds; and (2) oral dosing with a drug known to prevent sorbitol production - both metabolites disappeared. When normal mature lenses were incubated in 35.5 mM 13C1-glucose, both 13C1-lactate and 13C3-lactate were produced. Preservation of the 13C label at C1 is likely through the formation of 13C1-S3P and -F3P, which were then split through an aldolase-like mechanism into two 3-carbon compounds, one an unlabeled glycerol and the other 13C1-alpha-glycerophosphate (from S3P) and 13C1-dihydroxyacetone phosphate (from F3P). These reactions can contribute to the increase in alpha-glycerophosphate observed in both the streptozotocin-induced diabetic lenses and lenses incubated in high glucose.

Aging↗

Nuclear magnetic resonance studies of sugar metabolism in the human infant lens.

We investigated sugar metabolism in lenses isolated from infants with stage-5 retinopathy of prematurity (ROP). The lenses were incubated in 13C-labeled glucose (35.5 mM) or galactose (30 mM) for 24 h at 37.5 degrees C. Six to 8 lenses were pooled for each experiment. 13C-nuclear magnetic resonance spectroscopy was then performed on perchloric acid extracts of these lenses. Active aldose metabolism was indicated by the production of lactate. However, neither sorbitol nor dulcitol was detected. On the other hand, an experimental aldose reductase inhibitor, sorbinil, caused a decrease in the hexose monophosphate shunt activity suggesting an active polyol pathway. Both polyols therefore appeared to have been metabolized further. In addition, galactose metabolism was unique in that it not only involved a galactose-->glucose epimerization, but also a direct galactose-->ribose conversion, and a production of galactose-3-phosphate. Although it is possible that the ROP lenses have adapted to oxidative stress during the postnatal high-oxygen incubation for premature infants, we believe the observed sugar metabolism is representative of the developing human lens.

Aldehyde Reductase↗

Thermoregulatory vasoconstriction and perianesthetic heat transfer.

Heat transfer between the core and its environment in normothermic and slightly hypothermic situations is determined largely by the influence of vasomotion on convection. Tonic vasoconstriction, the normal barrier to heat loss from the core, is impaired upon induction of anesthesia. The resulting dilation of the arteriovenous shunts leads to redistribution of heat from the core to the periphery, diminishing the temperature gradient between the two compartments. With reemergence of thermoregulatory vasoconstriction at core temperatures near 34 degrees C, the core and the periphery are again separated, with metabolic heat being largely constrained to the core. Under normal conditions of mild thermal stress, thermoregulatory vasoconstriction is thus able to protect core temperature by reducing cutaneous heat transfer and functionally isolating the peripheral and core thermal compartments. Consequently, anesthetic-induced alterations in vasomotor tone is one of the major factors influencing core temperature in patients who are not actively cooled or warmed. In contrast, thermoregulatory tone is insufficient to prevent core temperature perturbations in patients undergoing vigorous cutaneous cooling or warming.

Anesthesia, General↗

Clinical research on acute hemoptysis treated with kexue ning.

The efficacy of Kexue Ning (stopping hemoptysis) in the treatment of acute hemoptysis, and its comparison with that of the control group (treated with western medicine) are reported in this paper. The results showed that both the time for cessation of hemoptysis and the cure rate were better in the therapeutic group than in the control group. In addition, marked improvement in blood rheology after treatment was also found in the therapeutic group, but not in the control group.

Acute Disease↗

The TNF receptor 1-associated protein TRADD signals cell death and NF-kappa B activation.

Many diverse activities of tumor necrosis factor (TNF) are signaled through TNF receptor 1 (TNFR1). We have identified a novel 34 kDa protein, designated TRADD, that specifically interacts with an intracellular domain of TNFR1 known to be essential for mediating programmed cell death. Overexpression of TRADD leads to two major TNF-induced responses, apoptosis and activation of NF-kappa B. The C-terminal 118 amino acids of TRADD are sufficient to trigger both of these activities and likewise sufficient for interaction with the death domain of TNFR1. TRADD-mediated cell death can be suppressed by the crmA gene, which encodes a specific inhibitor of the interleukin-1 beta-converting enzyme. However, NF-kappa B activation by TRADD is not inhibited by crmA expression, demonstrating that the signaling pathways for TNF-induced cell death and NF-kappa B activation are distinct.

3T3 Cells↗