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Biomedical subjects

T Jiang

Publications and source records attributed to T Jiang.

At least 19 recordsLinked to original sources

Crystal structure of allophycocyanin from red algae Porphyra yezoensis at 2.2-A resolution.

The crystal structure of allophycocyanin from red algae Porphyra yezoensis (APC-PY) at 2.2-A resolution has been determined by the molecular replacement method. The crystal belongs to space group R32 with cell parameters a = b = 105.3 A, c = 189.4 A, alpha = beta = 90 degrees, gamma = 120 degrees. After several cycles of refinement using program X-PLOR and model building based on the electron density map, the crystallographic R-factor converged to 19.3% (R-free factor is 26.9%) in the range of 10.0 to 2.2 A. The r.m.s. deviations of bond length and angles are 0.015 A and 2.9 degrees, respectively. In the crystal, two APC-PY trimers associate face to face into a hexamer. The assembly of two trimers within the hexamer is similar to that of C-phycocyanin (C-PC) and R-phycoerythrin (R-PE) hexamers, but the assembly tightness of the two trimers to the hexamer is not so high as that in C-PC and R-PE hexamers. The chromophore-protein interactions and possible pathway of energy transfer were discussed. Phycocyanobilin 1alpha84 of APC-PY forms 5 hydrogen bonds with 3 residues in subunit 2beta of another monomer. In R-PE and C-PC, chromophore 1alpha84 only forms 1 hydrogen bond with 2beta77 residue in subunit 2beta. This result may support and explain great spectrum difference exists between APC trimer and monomer.

Amino Acid Sequence

Structure and function of chromophores in R-Phycoerythrin at 1.9 A resolution.

The crystal structure of R-Phycoerythrin (R-PE) from Polysiphonia urceolata has been refined to a resolution of 1.9 A, based on the atomic coordinates of R-PE determined at 2.8 A resolution, through the use of difference Fourier method and steorochemistry parameters restrained refinement with model adjustment according to the electron density map. Crystallographic R-factor of the refined model is 0.195 (Rfree = 0.282) from 8-1.9 A. High resolution structure of R-PE showed precise interactions between the chromophores and protein residues, which explained the spectrum characteristic and function of chromophores. Four chiral atoms of phycourobilin (PUB) were identified as C(4)-S, C(16)-S, C(21)-S, and C(20)-R. In addition to the coupling distances of 19 A to 45 A between the chromophores which were observed and involved in the energy transfer pathway, high resolution structure of R-PE suggested other pathways of energy transfer, such as the ultrashort distance between alpha140a and beta155. It has been proposed that aromatic residues in linker proteins not only influence the conformation of chromophore, but may also bridge chromophores to improve the energy transfer efficiency.

Amino Acid Sequence

Membrane-permeant esters of phosphatidylinositol 3,4,5-trisphosphate.

Phosphoinositide 3-OH kinases and their products, D-3 phosphorylated phosphoinositides, are increasingly recognized as crucial elements in many signaling cascades. A reliable means to introduce these lipids into intact cells would be of great value for showing the physiological roles of this pathway and for testing the specificity of pharmacological inhibitors of the kinases. We have stereospecifically synthesized di-C8-PIP3/AM and di-C12-PIP3/AM, the heptakis(acetoxymethyl) esters of dioctanoyl- and dilauroylphosphatidylinositol 3,4,5-trisphosphate, in 14 steps from myo-inositol. The ability of these uncharged lipophilic derivatives to deliver phosphatidylinositol 3,4,5-trisphosphate across cell membranes was demonstrated on 3T3-L1 adipocytes and T84 colon carcinoma monolayers. Insulin stimulation of hexose uptake into adipocytes was inhibited by the kinase inhibitor wortmannin and was largely restored by di-C8-PIP3/AM, which had no effect in the absence of insulin. Thus phosphatidylinositol 3,4,5-trisphosphate or a metabolite was necessary but not sufficient for stimulation of hexose transport. In T84 epithelial monolayers, di-C12-PIP3/AM mimicked epidermal growth factor in inhibiting chloride secretion and potassium efflux, suggesting that phosphatidylinositol 3,4, 5-trisphosphate was sufficient to modulate these fluxes and mediate epidermal growth factor's action.

3T3 Cells

Differential regulation of astrocytic mRNAs in the rat striatum after lesions of the cortex or substantia nigra.

This study evaluates the time course of expression of three astrocytic mRNAs, glial fibrillary acidic protein (GFAP), apolipoprotein E (ApoE), and clusterin, in the rat striatum (ST) following a unilateral lesion of either the cortex (CX) or the substantia nigra (SN), using Northern blot and in situ hybridization analyses. We found that while there was a time-dependent increase in astrocytic GFAP mRNA in the deafferented ST following both the CX and the SN lesions, the time course of the response was different between the two lesion paradigms. Specifically, the increase in GFAP mRNA in striatal astrocytes after the SN lesion was rapid and transient returning to control levels by 10 days postlesion, while the response was long lasting and remained increased until at least 27 days after the CX lesion. In addition, the mRNA response for both ApoE and clusterin was differentially regulated in response to the two lesions. Specifically, both clusterin and ApoE mRNAs were rapidly increased in the ST following the CX lesion while both mRNAs remained unchanged following the SN lesion. Data from this study extend information derived from previous investigations on the multifunctional role of astrocytes in the response to brain injury. Specifically, our data support the notion that while the time course of the GFAP response in striatal astrocytes may vary between lesion paradigms, the upregulation of GFAP is part of a generalized response of reactive astrocytes to diverse brain injuries. By comparison, upregulation of the mRNAs for the lipoproteins clusterin and ApoE are lesion specific and may play a role in the transport of recycled myelin lipids from dying axons to actively growing axons and dendrites in reactive synaptogenesis.

Animals

The thrombin receptor elevates intracellular calcium in adult rat ventricular myocytes.

While there is evidence that thrombin receptor activation leads to contractile dysfunction and induces arrhythmias in ischemic/reperfused cardiac tissue, thrombin is variably reported to modulate intracellular calcium in cardiomyocytes. The present study demonstrates that thrombin receptor activation leads to a rise in intracellular calcium in adult ventricular myocytes and serves to reconcile previous discrepant findings. The thrombin receptor-derived agonist peptide (SFLLRN, a portion of the tethered ligand created by thrombin's proteolytic actions) increases cytosolic calcium and twitch amplitude in cardiomyocytes isolated from adult ventricles. The truncated control peptide FLLRN has no effect, establishing that the response to SFLLRN results from a specific agonist peptide-receptor interaction. However, the response to SFLLRN occurs only at high agonist peptide concentrations and thrombin itself is inactive. This result is not compatible with an action of SFLLRN at a distinct protease-activated receptor (PAR-2; which is activated by SFLLRN, but not by thrombin), since SLIGRL (a ligand which is selective for PAR-2, but not the thrombin receptor) has no effect. Rather, the enzyme-based cell isolation procedure may partially cleave the thrombin receptor and influence cell responses, since concentrations of SFLLRN which are sub-threshold in enzymatically disaggregated myocytes significantly increase the force of isometric contraction of intact rat papillary muscles. These studies provide the first evidence that thrombin receptor activation leads to a change in intracellular calcium and a positive inotropic response in adult ventricular myocardium.

Animals

D-myo-Inositol 1,4,5,6-tetrakisphosphate produced in human intestinal epithelial cells in response to Salmonella invasion inhibits phosphoinositide 3-kinase signaling pathways.

Several inositol-containing compounds play key roles in receptor-mediated cell signaling events. Here, we describe a function for a specific inositol polyphosphate, D-myo-inositol 1,4,5,6-tetrakisphosphate [Ins(1,4,5,6)P4], that is produced acutely in response to a receptor-independent process. Thus, infection of intestinal epithelial cells with the enteric pathogen Salmonella, but not with other invasive bacteria, induced a multifold increase in Ins(1,4,5,6)P4 levels. To define a specific function of Ins(1,4,5,6)P4, a membrane-permeant, hydrolyzable ester was used to deliver it to the intracellular compartment, where it antagonized epidermal growth factor (EGF)-induced inhibition of calcium-mediated chloride (Cl-) secretion (CaMCS) in intestinal epithelia. This EGF function is likely mediated through a phosphoinositide 3-kinase (PtdIns3K)-dependent mechanism because the EGF effects are abolished by wortmannin, and three different membrane-permeant esters of the PtdIns3K product phosphatidylinositol 3,4,5-trisphosphate mimicked the EGF effect on CaMCS. We further demonstrate that Ins(1,4,5,6)P4 antagonized EGF signaling downstream of PtdIns3K because Ins(1,4,5, 6)P4 interfered with the PtdInsP3 effect on CaMCS without affecting PtdIns3K activity. Thus, elevation of Ins(1,4,5,6)P4 in Salmonella-infected epithelia may promote Cl- flux by antagonizing EGF inhibition mediated through PtdIns3K and PtdInsP3.

Epidermal Growth Factor

Upregulation of stathmin (p19) gene expression in adult rat brain during injury-induced synapse formation.

Stathmin (p19) is developmentally regulated as a neural-enriched phosphoprotein associated with neurite outgrowth and synaptic formation during cell proliferation and differentiation, and remains highly abundant in adult rat brain. Whether stathmin is involved in injury-induced reactive synaptogenesis in adult rat was examined in this study. Following unilateral cortical lesion, a significant increase in stathmin mRNA expression was found in the cells of contralateral homotypic cortex and in the subventricular zone of the lateral ventricle. This increase coincided in time with the corticostriatal axon sprouting and synaptic remodeling previously found in denervated striatum. Our data suggest that stathmin plays an important role in regulation of reactive synaptogenesis in adult brain.

Animals

Expression of protein kinase C beta in the heart causes hypertrophy in adult mice and sudden death in neonates.

Protein kinase C (PKC) activation in the heart has been linked to a hypertrophic phenotype and to processes that influence contractile function. To establish whether PKC activation is sufficient to induce an abnormal phenotype, PKCbeta was conditionally expressed in cardiomyocytes of transgenic mice. Transgene expression in adults caused mild and progressive ventricular hypertrophy associated with impaired diastolic relaxation, whereas expression in newborns caused sudden death associated with marked abnormalities in the regulation of intracellular calcium. Thus, the PKC signaling pathway in cardiocytes has different effects depending on the timing of expression and, in the adult, is sufficient to induce pathologic hypertrophy.

Age Factors

Functional consequences of acute collagen degradation studied in crystalloid perfused rat hearts.

OBJECTIVES: The impact of acute collagen disruption by the disulfide donor, 5,5'-dithio-2-nitrobenzoic acid (DTNB) on ventricular properties was tested in rat hearts. METHODS: Collagen was degraded acutely in 13 isolated, isovolumically contracting rat hearts by perfusion with 1 mM DTNB added to Krebs-Henseleit solution for 1 hour followed by 2-hour perfusion with normal solution. Another 13 hearts were perfused with normal solution for 3 hours (Control). RESULTS: Collagen content was 3.5 +/- 0.5% of ventricular dry weight in control group compared with 2.1 +/- 0.4% in DTNB group (decrease by 40%, p < 0.01). Scanning electron micrographs revealed loss of the delicate collagen network surrounding muscle fibers in DTNB treated hearts. Developed pressure at a fixed volume decreased to 86 +/- 17% of the baseline value after 3-hour perfusion in the control group, whereas in DTNB treated hearts developed pressure fell to 68 +/- 13% (p < 0.01). End-diastolic pressure was set at 5 mmHg at the beginning of the experiment and rose to 15 +/- 8 mmHg in control and 30 +/- 13 mmHg (p < 0.01) in the treated hearts. Concomitantly, wet-to-dry weight ratio increased from 5.63 +/- 0.26 in control to 6.07 +/- 0.11 (p < 0.05) in the DTNB treated hearts. A separate set of experiments on isolated myocytes excluded the possibility of a direct effect of DTNB on myocyte contractile function. CONCLUSIONS: These data suggested that with 40% collagen disruption by DTNB there is a significant increase in tissue edema that results in a decrease in chamber capacitance; in addition, there is a significant decrease in systolic performance which reflects the combined effect of edema and loss of collagen.

Animals

Facial and autonomic responses to biological and artificial olfactory stimuli in human neonates: re-examining early hedonic discrimination of odors.

Responses of awake and asleep 3-day-old human neonates were recorded to the presentation of artificial (vanillin, butyric acid, formula milks) and biological (breast milk, amniotic fluid) odorants matched on subjective intensity and trigeminal dimensions. The responses included behavioral (facial and oral movements) and autonomic (respiration, differential skin temperature) measures. The neonates reacted with significant facial and respiratory changes to low concentrations of olfactory stimuli during the various behavioral states. The analysis of olfacto-facial configurations revealed that behavioral markers of disgust (nose wrinkling, upper lip raising) discriminated between some odors judged as being pleasant and unpleasant by adult raters (vanillin vs. butyric acid). However, although some early predisposition to process the affective significance of stimuli may be suggested, no convincing evidence was obtained that neonates discriminated the hedonic valence of odors within the same perceptual space as adults. Finally, neonates evinced a differential pattern of respiratory responding to the presentation of milk odors according to the mode of feeding (breast vs. bottle feeding), suggesting that early olfactory discrimination may be mediated by stimuli with high ecological salience.

Autonomic Nervous System

Synergies and coincidence requirements between NO, cGMP, and Ca2+ in the induction of cerebellar long-term depression.

Parallel fiber synapses onto Purkinje neurons in acute cerebellar slices undergo long-term depression (LTD) when presynaptic activity coincides with postsynaptic depolarization. These electrical inputs can be respectively replaced by nitric oxide (NO) and Ca2+ photolytically released inside the Purkinje neuron, showing that these two messengers are sufficient for LTD induction. NO acts via cGMP production because inhibitors of guanylate cyclase prevent LTD but can be circumvented by photoreleased cGMP combined with Ca2+ elevation. Three inhibitors of cGMP-dependent protein kinase, Rp-8Br-PET-cGMPS, KT5823, and a novel pseudosubstrate peptide, all block LTD. LTD induction permits <10 ms gap between NO release and Ca2+ elevation, whereas 200-300 ms is allowed between uncaged cGMP and Ca2+ increase. This surprising difference in timing precision can be explained either by tighter localization and faster decay of cGMP when generated by NO rather than uncaging, or by two independent coincidence detectors in series.

Animals

Modification of colonic fermentation by bifidobacteria and pH in vitro. Impact on lactose metabolism, short-chain fatty acid, and lactate production.

Colonic fermentation plays an important role in the prevention of lactose intolerance and intestinal disorders. The objectives of this study were to evaluate whether supplementation with bifidobacteria modify colonic fermentation of lactose and short-chain fatty acid production and to assess influence of the pH in an in vitro continuous culture system. There was a significantly greater reduction in lactose concentrations at pH 6.7 than that at either pH 6.2 or pH 5.7, accompanied by the highest beta-galactosidase activity and D-lactate production. Bifidus supplementation reduced lactose and D-lactate concentrations and increased acetate production at pH 6.7. The study demonstrates that lactose is rapidly metabolized by colonic bacteria and lactose fermentation in vitro is pH dependent with a maximum rate at pH 6.7. Bifidobacteria supplementation may have the potential to improve lactose fermentation and to manipulate SCFA and lactate production.

Adult

Surveillance for polio eradication in the People's Republic of China.

A case-based virus surveillance system for wild poliovirus in China was developed. By 1993, all 30 provincial immunization units and, by 1994, all 29 provincial laboratories were sending computerized data to the national level. In 1993, a county-level, computerized map was operationalized that permitted visual monitoring of the progress of the polio eradication program every month by county. In 1993, wild poliovirus type 1 was detected in 8 provinces. Wild poliovirus mainly caused clusters of polio cases identified by a surveillance system that detected primarily clinical polio in children <5 years old (1 stool sample was collected on approximately 50% of reported cases). By 1995, the surveillance system had reached certification-like levels (80% of acute flaccid paralysis [AFP] patients with 2 stool specimens and AFP case rate of 1/100,000 children <15 years old). No indigenous wild poliovirus was detected in 1995. This general case-based model can be applied to measles and other important diseases, and may then lead to a more rapid decrease in adverse health outcomes.

Adolescent

In vitro lactose fermentation by human colonic bacteria is modified by Lactobacillus acidophilus supplementation.

Adaptation of the colonic flora to lactose may contribute to lactose digestion in lactose maldigesters, and supplementation with Lactobacillus acidophilus may modify colonic fermentation of lactose and short-chain fatty acid production. We evaluated the capability of colonic bacteria to ferment lactose and the ability of L. acidophilus to modify lactose fermentation by the colonic microflora in vitro. An anaerobic continuous culture was established and inoculated with fresh samples of human feces. Lactose infusion was maintained at 25 g/d and pH at 6.7. L. acidophilus strain LA-1 (1.5 x 10(10) cells) was introduced into the fermenter on d 0 or added daily on d 0 through 4. The control was the continuous culture without the addition of lactobacilli. Rapid adaptation of colonic bacteria to lactose occurred within 1-2 d, with a significant decrease in lactose concentration and increase in beta-galactosidase activity, and lactose concentrations fell below 3 mmol/L by d 7. Supplementation with strain LA-1 resulted in a significantly greater decrease in lactose concentration and greater increase in acetate and propionate production within the first day compared with the control group. However, there was no significant difference between the fermentation treated with L. acidophilus daily and the control after the first day. These data suggest that the colonic bacteria adapt quickly to lactose, causing efficient utilization of lactose. L. acidophilus supplementation may enhance lactose fermentation during early periods when the adaptation is not established in this model.

Adult

Beta 2-adrenergic receptors enhance contractility by stimulating HCO3(-)-dependent intracellular alkalinization.

Previous studies established that beta 2-adrenergic receptors enhance the amplitude, without abbreviating the kinetics, of the twitch in adult rat ventricular myocytes. The present study was designed to identify the dominant signaling mechanism mediating this response. Myocytes from adult rat ventricles were loaded with the pH-sensitive fluorophore 2',7'-bis(carboxyethyl)-5(6')-carboxyfluorescein, and simultaneous measurements of intracellular pH and contraction were performed during electrical field stimulation under basal conditions and after stimulation with isoproterenol or the selective beta 2-receptor agonist zinterol. Inhibition of protein kinase A with H-7 completely inhibited the isoproterenol-dependent, but not the zinterol-dependent, positive inotropic response. The effect of zinterol to increase twitch amplitude was associated with an alkalinization of 0.07 +/- 0.02 pH unit, which was not prevented by inhibition of the Na+/H+ exchanger with hexamethylene-amiloride. Rather, removal of bicarbonate from the extracellular buffer prevented the beta 2-receptor-dependent alkalinization as well as the positive inotropic response. These results indicate that beta 2-adrenergic receptors induce a positive inotropic response in adult rat ventricular myocytes via a adenosine 3',5'-cyclic monophosphate-independent mechanism that involves intracellular alkalinization due to activation of a bicarbonate-dependent intracellular pH regulatory mechanism.

Alkalies

Improvement of lactose digestion by humans following ingestion of unfermented acidophilus milk: influence of bile sensitivity, lactose transport, and acid tolerance of Lactobacillus acidophilus.

The influence of bile sensitivity, lactose transport, and acid tolerance of Lactobacillus acidophilus on in vivo digestion of lactose was investigated. Four strains of L. acidophilus exhibiting varied degrees of lactose transport, beta-galactosidase activity, and bile sensitivity were used to prepare unfermented acidophilus milks. Lactose malabsorption was evaluated by measuring breath H2 excretion off 11 lactose maldigesting subjects following ingestion of four acidophilus test milks. Test meals were fed in a randomized double-blind protocol. Consumption of acidophilus milk (2% fat) containing strains B, N1, and E significantly reduced mean total H2 production compared with that of the control reduced-fat (2% fat) milk, but milk containing strain ATCC 4356 did not differ from the control. Acidophilus milk containing L. acidophilus N1 was the most effective of the four acidophilus milks in improving lactose digestion and tolerance. Strain N1 exhibited the lowest beta-galactosidase activity and lactose transport but the greatest bile and acid tolerance of the four strains. The results indicated that bile and acid tolerance may be important factors to consider when L. acidophilus strains are selected for improving lactose digestion and tolerance.

Adult

Crystal structure of R-phycoerythrin from Polysiphonia urceolata at 2.8 A resolution.

The structure of R-phycoerythrin (R-PE) from Polysiphonia urceolata was determined at 2.8 A resolution. The crystals belong to space group R3 with unit cell dimensions of a = b = 189.8 A, c = 60.1 A. The subunit composition of R-PE is (alpha 2 beta 2)3 gamma. The three-dimensional structure of R-PE was solved by the multiple isomorphous replacement method. After several cycles of model building and refinement, the crystallographic R-factor of the final model is 18.0% with data from 10.0 to 2.8 A resolution. The four phycoerythrobilin chromophores alpha 84, alpha 140a, beta 84 and beta 155 in an (alpha beta) unit are each covalently bound to a cysteine residue through ring A. The phycourobilin chromophore is bound to cysteine beta 50 by ring A and bound to cysteine beta 61 by ring D. The ring A and ring D of phycourobilin deviate from the conjugate plane formed by ring B and ring C and the four rings form a boat-shaped structure. R-PE contains a 34 kDa gamma subunit that is assumed to lie in the central channel of the molecular disc (alpha 2 beta 2)3. The energy transfer and relationship between cysteine residues and chromophores are discussed.

Bacterial Proteins