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Biomedical subjects

J Zhang

Publications and source records attributed to J Zhang.

At least 469 records · Page 26Linked to original sources

Hydropassive evidence and effective factors in stomatal oscillations of Glycyrrhiza inflata under desert conditions.

Whether stomata oscillations induced by atmospheric drought stress are hydropassive or metabolic energy-dependent and the thresholds of some effective factors were studied in Glycyrrhiza inflata. The metabolic inhibitors NaN(3) and carbonyl cyanide-m-phenyl-hydrazone inhibited the respiration. However they could not significantly change the intensity (amplitude/average) of the stomata oscillation. The leaf turgor-pressure was fluctuated simultaneously with the stomata oscillations, whereas the K(+) content of the guard cells did not show oscillations. The oscillation intensity was found to be regulated by vapour pressure deficit (VPD), the proportion of the retained root and the stem flux lag. The minimum threshold of VPD, roots and the stem flux lag may be required to induce the stomata oscillations. The fluctuations of the leaf turgor-pressure induced by the non-synchronization between the transpiration demand and provide of water by the stem flux may be the direct cause of the stomata oscillations in G. inflata under the conditions of high transpiration demand.

Journal Article↗

Peripheral interleukin-6 administration increases extracellular concentrations of serotonin and the evoked release of serotonin in the rat striatum.

Previous studies have indicated that peripheral administration of interleukin-6 (IL-6) increases brain concentrations of tryptophan and 5-hydroxyindoleacetic acid (5-HIAA), the major catabolite of serotonin (5-HT). To determine whether these changes were related to increased synaptic release of 5-HT, we studied the responses to peripheral administration of IL-6 by in vivo microdialysis and in vivo amperometry. Intraperitoneal injection of recombinant IL-6 resulted in an elevation of microdialysate concentrations of 5-HT in the rat striatum. Also, amperometric measurements indicated that i.p. IL-6 enhanced the 5-HT-like signal obtained from the striatum following electrical stimulation of the dorsal raphe nucleus. These results indicate that the increases in brain concentrations of 5-HIAA observed in earlier studies indeed reflect increased synaptic release of 5-HT.

Animals↗

Ozone exposure thresholds and foliar injury on forest plants in Switzerland.

Canton Ticino in southern Switzerland is exposed to some of the highest concentrations of tropospheric ozone in Europe. During recent field surveys in Canton Ticino, foliar symptoms identical to those caused by ozone have been documented on native tree and shrub species. In Europe, the critical ozone level for forest trees has been defined at an AOT40 of 10 ppm.h O3 (10 ppm.h accumulated exposure of ozone over a threshold of 40 ppb) during daylight hours over a six-month growing season. The objective of this study was to determine the amount of ambient ozone required to induce visible foliar symptoms on various forest plant species in southern Switzerland. Species were grown within eight open-top chambers and four open plots at the Vivaio Lattecaldo Cantonal Forest Nursery in Ticino, Switzerland. Species differed significantly in terms of the ppb.h exposures needed to cause visible symptoms. The most to least symptomatic species grown within open-plots in this study rank as Prunus serotina, Salix viminalis, Vibrnum lantana, Rhamnus cathartica, Betula pendula, Rumex obtusifolius, Sambucus racemosa, Morus nigra, Prunus avium, Fraxinus excelsior, Rhamnus frangula, Alnus viridis, Fagus sylvatica and Acer pseudoplatanus. Similar rankings were obtained in the non-filtered chamber plots. The ranking of species sensitivity closely follows AOT values for the occurrence of initial symptoms and symptom progression across the remainder of the exposure season. Species that first showed evidence of foliar injury also demonstrated the most sensitivity throughout the growing season, with symptoms rapidly advancing over ca. 25-30% of the total plant leaf surfaces by the end of the observation period. Conversely, those species that developed symptoms later in the season had far less total injury to plant foliage by the end of the observation period (1.5 to < 5% total leaf area injured). The current European ambient ozone standard may be insufficient to protect native plant species from visible foliar injury, and many more native species may be sensitive to ozone-induced foliar injury than are currently known.

Environmental Monitoring↗

Variation of gas exchange within native plant species of Switzerland and relationships with ozone injury: an open-top experiment.

Gas exchange and ozone-induced foliar injury were intensively measured during a 6-day period in mid-August 1998 on leaves of Acer pseudoplatanus, Betula pendula, Corylus avellana, Fagus sylvatica, Fraxinus excelsior, Morus nigra, Prunus avium, Prunus serotina, Rhamnus cathartica, and Viburnum lantana at a forest nursery site in Canton Ticino, Switzerland. Plants were grown in four open plots (AA), four open-top chambers receiving carbon-filtered (CF) air, and four receiving non-filtered (NF) air. Significant variation in gas exchange (F > 12.7, P < 0.001) was detected among species with average net photosynthesis and average stomatal conductance differing by a factor of two. Species also varied significantly in foliar injury for those leaves for which we measured gas exchange (F = 39.6, P < 0.001). Fraxinus excelsior, M. nigra, P. avium, P. serotina, R. cathartica, and V. lantana showed more injury than A. pseudoplatanus, B. pendula, C. avellana, and Fagus sylvatica. Plants grown in CF chambers had significantly higher net photosynthesis (A) and stomatal conductance to water vapor (gwv), and lower foliar injury than plants grown in NF chambers and AA plots; interactions between species and ozone treatments were significant for all variables (F > or = 2.2, P < 0.05) except gwv (F = 0.7, P > 0.1). Although A and gwv decreased and foliar injury increased with leaf age, the magnitude of these changes was lower for plants grown in CF chambers than for plants grown in NF chambers and AA plots. Neither ozone uptake threshold (r = 0.26, P > 0.20) nor whole-plant injury (r = -0.15, P > 0.41) was significantly correlated with stomatal conductance across these species. It appears that the relationships between stomatal conductance and foliar injury are species-specific and interactions between physiology and environments and leaf biochemical processes must be considered in determining species sensitivity to ambient ozone exposures.

Air Pollution↗

Protein kinase C, rather than protein kinase A is involved in follicle-stimulating hormone-mediated meiotic resumption of mouse cumulus cell-enclosed oocytes in hypoxanthine-supplemented medium.

It has been reported that protein kinase C (PKC) activation participated in the porcine and bovine oocyte maturation, but not in mouse oocyte maturation in vitro. In the present study, the activators and inhibitors of protein kinase A (PKA) (forskolin, CDPKI and MDL-12230A) or PKC (PMA, staurosporine and sphingosine) were used to investigate the in vitro effect of PKA or PKC on spontaneous murine oocyte maturation, oocyte resumption of meiosis from HX inhibiting medium (medium+HX), and follicle stimulating hormone (FSH)-induced oocyte maturation. The results showed that when cumulus cell enclosed oocytes (CEOs) or denuded oocytes (DOs) were cultured for 24 h in the medium supplemented with forskolin (5 microM), an activator of adenylate cyclase, the spontaneous oocyte maturation were inhibited. A transient exposure (2 h) to forskolin (2-10 microM) in the medium+HX, and then transferred to a new medium+HX for the further culture, stimulated CEO resumption of meiosis. CDPKI (10(-10)-10(-6) M), an inhibitor of PKA, also stimulated oocyte meiotic maturation of CEO in the medium+HX, but not on DO. However, MDL-12230A (10(-12)-10(-9) M), an inhibitor of adenylate cyclase, did not promote oocyte maturation in HX arrested CEO. CDPKI (10(-10)-10(-6) M) or MDL-12230A (10(-12)-10(-9) M) had no effect on FSH-stimulated oocyte meiotic resumption, except at high doses of CDPKI (10(-7)-10(-6) M) or MDL-12230A (10(-9) M) which inhibited the FSH-induced formation of the first polar body (PB1). An activator of PKC, PMA (10(-11)-10(-7) M) dose-dependently inhibited spontaneous oocyte maturation of CEO or DO. Inhibitors of PKC, staurosporine (10(-9)-10(-6) M) or sphingosine (10(-8)-10(-5) M) induced oocytes in CEOs to resume meiosis in the presence of HX in a dose dependent manner, but had no effect on DOs. FSH (50IU/L) stimulated mouse oocytes in CEOs to override the arrest of HX and resume meiosis, while PMA, at the level of 10(-8)-10(-6) M, dramatically inhibited the stimulatory effect of FSH. These results indicate that PKC or PKA may be implicated in the regulation of mouse oocyte maturation. Thus while sustained high level of cAMP or PKA inhibit the resumption of meiosis, a transient rise in cAMP or PKA levels promotes oocyte maturation. The activation of PKC can also block oocyte meiotic resumption. Thus the inactivation of PKC, instead of the transient rise of PKA activity, appears to be involved in the process of FSH-mediated oocyte meiotic maturation.

Animals↗

Synergistic neurotoxicity of opioids and human immunodeficiency virus-1 Tat protein in striatal neurons in vitro.

Human immunodeficiency virus (HIV) infection selectively targets the striatum, a region rich in opioid receptor-expressing neural cells, resulting in gliosis and neuronal losses. Opioids can be neuroprotective or can promote neurodegeneration. To determine whether opioids modify the response of neurons to human immunodeficiency virus type 1 (HIV-1) Tat protein-induced neurotoxicity, neural cell cultures from mouse striatum were initially characterized for mu and/or kappa opioid receptor immunoreactivity. These cultures were continuously treated with morphine, the opioid antagonist naloxone, and/or HIV-1 Tat (1-72) protein, a non-neurotoxic HIV-1 Tat deletion mutant (TatDelta31-61) protein, or immunoneutralized HIV-1 Tat (1-72) protein. Neuronal and astrocyte viability was examined by ethidium monoazide exclusion, and by apoptotic changes in nuclear heterochromatin using Hoechst 33342. Morphine (10nM, 100nM or 1microM) significantly increased Tat-induced (100 or 200nM) neuronal losses by about two-fold at 24h following exposure. The synergistic effects of morphine and Tat were prevented by naloxone (3microM), indicating the involvement of opioid receptors. Furthermore, morphine was not toxic when combined with mutant Tat or immunoneutralized Tat. Neuronal losses were accompanied by chromatin condensation and pyknosis. Astrocyte viability was unaffected. These findings demonstrate that acute opioid exposure can exacerbate the neurodegenerative effect of HIV-1 Tat protein in striatal neurons, and infer a means by which opioids may hasten the progression of HIV-associated dementia.

Animals↗

Generation of a prostate epithelial cell-specific Cre transgenic mouse model for tissue-specific gene ablation.

To facilitate the elucidation of the genetic events that may play an important role in the development or tumorigenesis of the prostate gland, we have generated a transgenic mouse line with prostate-specific expression of Cre recombinase. This line, named PB-Cre4, carries the Cre gene under the control of a composite promoter, ARR2PB which is a derivative of the rat prostate-specific probasin (PB) promoter. Based on RT-PCR detection of Cre mRNA in PB-Cre4 mice or Cre-mediated activation of LacZ activity in PB-Cre4/R26R double transgenic mice, it is conclusively demonstrated that Cre expression is post-natal and prostatic epithelium-specific. Although the Cre recombination is detected in all lobes of the mouse prostate, there is a significant difference in expression levels between the lobes, being highest in the lateral lobe, followed by the ventral, and then the dorsal and anterior lobes. Besides the prostate gland, no other tissues of the adult PB-Cre4 mice demonstrate significant Cre expression, except for a few scattered areas in the gonads and the stroma of the seminal vesicle. By crossing the PB-Cre4 animals with floxed RXRalpha allelic mice, we demonstrate that mice, whose conventional knockout of this gene is lethal in embryogenesis, could be propagated with selective inactivation of RXRalpha in the prostate. Taken together, the results show that the PB-Cre4 mice have high levels of Cre expression and a high penetrance in the prostatic epithelium. The PB-Cre4 mice will be a useful resource for genetic-based studies on prostate development and prostatic disease.

Alleles↗

Degumming of ramie fibers by alkalophilic bacteria and their polysaccharide-degrading enzymes.

Three strains of alkalophilic bacteria, Bacillus sp. NT-39, NT-53 and NT-76, were selected for the degumming of ramie fibers and production of polysaccharide-degrading enzymes. After 48 h of incubation with the strains, the loss of the gum might amount to 5.0% or more of the fibers and a number of polysaccharide-degrading enzymes were secreted to the culture supernatants. The residual gum of the fibers decreased to 9.4% after 5 h of enzymatic degumming. Analysis of gum contents and enzyme activities revealed that pectate lyase and xylanase played an important role in the degradation of residual gum. Enzymatic degumming resulted in an increment of 5.4 ISO units in fiber brightness, whereas the reduction in bundle breaking tenacity of the fibers was less than 5.%. The results confirmed that degumming of ramie fibers by alkalophilic bacteria and their enzymes had substantial advantages.

Bacillus↗

Identification of inhibitors of heparin-growth factor interactions from combinatorial libraries of four-component condensation reactions.

Chemical libraries based on four-component condensation (4CC) reactions of isocyanides were constructed to identify compounds capable of blocking heparin binding to vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF). The reaction products in the synthesized libraries contain heparin mimetic functional groups such as carbohydrates, sulfonates, carboxylates, and hydroxy groups. These libraries have been screened for the inhibition of heparin binding to growth factors such as VEGF and bFGF. Single point screening at 5.0 microM of the 18,720 reaction products generated 26 candidates. The IC50S of these 26 compounds were determined using HPLC-purified products and 20 of the 26 showed significant inhibition of heparin binding to VEGF and/or bFGF. Eighteen of the 20 confirmed active compounds have a linear extended structure. Structures identified in this library revealed an initial relationship of structure and activity, thus providing direction for further investigation of this type of heparin mimetic libraries.

Chromatography, High Pressure Liquid↗

Synthesis and biodistribution of a new 99mTc nitrido complex for brain imaging.

Bis(N-isobutyl-dithiocarbamato) nitrido technetium-99m complex [99mTcN(IBDTC)2] (IBDTC: N-isobutyl dithiocarbamato) was synthesized by the reduction of 99mTcO4- into [99mTc = N]2+ with stannous chloride in the presence of succinic dihydrazide and propylenediamine tetraacetic acid, followed by the addition of sodium N-isobutyl dithiocarbamate dihydrate. The radiochemical purity of the complex was over 90% as measured by thin layer chromatography. It was stable over 6h at room temperature. Its partition coefficient indicated that it is a good lipophilic complex. Biodistribution in mice demonstrated that the complex accumulated in the brain with high uptake and good retention. The brain uptake (ID%/g) was 6.22, 5.45 and 3.88 and the brain/blood ratio was 1.51, 2.24, 1.84 at 5, 30 and 60 min post-injection, respectively. These results suggest potential usefulness of the complex as a brain perfusion imaging agent.

Animals↗

Preparation of 99mTcN(CBDTC)2 and its biodistribution in mice.

In order to seek a new class of brain perfusion imaging agent containing [99mTcN]2+ core, bis(N-cyclobutyl dithiocarbamato) nitrido technetium-99m complex [99mTcN(CBDTC)2] (CBDTC: N-cyclobutyl dithiocarbamato) was synthesized through a simple and efficient method, which can be utilized for the preparation of a radiopharmaceutical through a lyophilized kit formulation. The radiochemical purity of the complex was over 90%, as measured by thin layer chromatography. No decomposition of the complex at room temperature was observed over a period of 6 h. It's partition coefficient indicated that it is a good lipophilic complex. Biodistribution in mice showed that the complex was significantly retained in the brain. The brain uptake (ID %/g) was 3.61, 3.15 and 2.62 and the brain/blood ratio was 1.00, 1.44 and 1.30 at 5, 30 and 60 min post-injection, respectively. These results suggest that this compound could be a potential brain perfusion imaging agent.

Animals↗

Monomeric IgE stimulates signaling pathways in mast cells that lead to cytokine production and cell survival.

Although IgE binding to mast cells is thought to be a passive presensitization step, we demonstrate herein that monomeric IgE (mIgE) in the absence of antigen (Ag) stimulates multiple phosphorylation events in normal murine bone marrow-derived mast cells (BMMCs). While mIgE does not induce degranulation or leukotriene synthesis, it leads to a more potent production of cytokines than IgE + Ag. Moreover, mIgE prevents the apoptosis of cytokine-deprived BMMCs, likely by maintaining Bcl-X(L) levels and producing autocrine-acting cytokines. The addition of Ag does not increase this IgE-induced survival. Since IgE concentrations as low as 0.1 microg/ml enhance BMMC survival, elevated plasma IgE levels in humans with atopic disorders may contribute to the elevated mast cell numbers seen in these individuals.

Animals↗

K-Cl co-transport: immunocytochemical and functional evidence for more than one KCC isoform in high K and low K sheep erythrocytes.

K-Cl co-transport (COT) is significantly higher in low K (LK), L-antigen (L) positive, than in high K (HK), M-antigen (M) positive, sheep red blood cells (SRBCs) and is inhibited by sheep allo-anti-L1 antibody. To answer the question of whether this difference in K-Cl co-transport activity resides at the level of the transporter or its regulation, a combined immunocytochemical and functional approach was taken. At least four isoforms of K-Cl COT encoded by different KCC genes are known, with 12 transmembrane domains and cytoplasmic C- and N-terminal domains (Ctd and Ntd, respectively). Polyclonal anti-rat (rt)KCC1 antibodies against a fusion peptide with 77 amino acids from the Ctd of rtKCC1 and anti-human (h)KCC3 against an 18-aa peptide from the Ntd of hKCC3, were prepared in rabbits (rb). Two distinctly separate protein bands of 180 and 145 kDa molecular mass were detected in hemoglobin-free ghosts from RBCs of two LK (one homozygous LL and one heterozygous LM) and one HK (homozygous MM) sheep by Western blots with rb anti-rtKCC1 and rb anti-hKCC3. Confocal microscopy showed specific immunostaining of KCC1 with rb anti-rtKCC1, and of KCC3 with rb anti-hKCC3, in white ghosts from both LK and HK SRBCs. To test the functional heterogeneity of K-Cl COT, the effect of the anti-L1 antibody was assessed on K-Cl COT activated by the kinase inhibitor staurosporine. Incubation of LK SRBCs with anti-L1 serum inhibited by 30% staurosporine-stimulated K-Cl COT suggesting that approximately two-thirds of the transport activity is independent of the L1 antigen. That staurosporine altered the L1 antigen/antibody reaction is unlikely since the action of another antibody, anti-Lp, stimulating the Na/K pump flux, was not modified. The present results, in conjunction with earlier work, lead to the hypothesis that the partial anti-L1 inhibition of K-Cl COT may be related to the molecular KCC dimorphism, seen in these cells with anti-KCC1 and anti-KCC3 antibodies.

Animals↗

Cl(-)-ATPases: biological active transporters.

Five widely documented mechanisms of chloride transport across plasma membranes are: anion-coupled antiport; sodium and hydrogen-coupled symport; Cl- channels; and an electrochemical coupling process. No genetic evidence has yet been provided for primary active chloride transport despite numerous reports of cellular Cl(-)-stimulated ATPases co-existing, in the same tissue, with uphill chloride transport that could not be accounted for by the five common chloride transport processes. Cl(-)-stimulated ATPase activity is a common property of practically all biological cells with the major location being of mitochondrial origin. It also appears that plasma membranes are sites of Cl(-)-stimulated ATPase activity. Recent studies of Cl(-)-stimulated ATPase activity and active chloride transport in the same membrane system, including liposomes, suggest a mediation by the ATPase in net movement of chloride up its electrochemical gradient across plasma membranes. Further studies, especially from a molecular biological perspective, are required to confirm a direct transport role to plasma membrane-localized Cl(-)-stimulated ATPases.

Adenosine Triphosphatases↗

Ethanol activates NFkappaB DNA binding and p56lck protein tyrosine kinase in human osteoblast-like cells.

Alcoholics frequently suffer from moderate to severe bone loss that results in bone fractures. Both decreased bone production and increased bone resorption have been postulated to contribute to ethanol (ETOH)-mediated bone loss. Bone resorption is induced by several proinflammatory cytokines such as interleukin-1 and -6. The expression of these cytokines is induced by the transcription factor NFkappaB, which, in turn, is activated by several kinases. It follows that protein kinase and NFkappaB activation may contribute to ETOH-induced bone loss. Accordingly, we sought to determine if ETOH activates protein tyrosine kinases (PTK) and NFkappaB DNA binding in a human osteoblast-like cell line (HOBIT). Ethanol at 50 and 100 mmol/L (reflective of blood ethanol levels reached in chronic alcoholics) for 24 h did not alter HOBIT cell viability. In contrast, 200 mmol/L ethanol decreased cell viability by 40%. Treatment of HOBIT cells with 100 mmol/L ETOH induced nuclear NFkappaB:DNA complex formation and NFkappaB activity. Incubation of HOBIT cells with ETOH at 50 and 100 mmol/L for 30 min induced a 2.5- and 4.2-fold increase in PTK activity, respectively. Preincubation of HOBIT cells with damnacanthal (DAM), which inhibits p56lck, blocked ETOH-mediated PTK activity; whereas, preincubation with herbimycin A, which inhibits pp60src, did not. DAM inhibited both ethanol-induced NFkappaB activation in HOBIT cells and interleukin-6 expression in primary human osteoblasts. Finally, preincubation with the protein kinase C inhibitor, bisindolylmaleimide I HCl (BIS), diminished ETOH-mediated PTK activity; whereas, preincubation with the protein kinase A inhibitor, H89, did not. These data demonstrate that ETOH induces NFkappaB nuclear translocation through p56lck in HOBIT cells. BIS' inhibition of PTK activation suggests that ETOH activates PTK through a protein kinase C-dependent pathway. These data suggest that ETOH may contribute to bone loss through activation of signal transduction that results in production of an osteoclastogenic cytokine (i.e., interleukin-6) in osteoblasts.

Alcoholism↗

Recombinant human extracellular matrix protein 1 inhibits alkaline phosphatase activity and mineralization of mouse embryonic metatarsals in vitro.

Two mRNAs are transcribed from the extracellular matrix protein 1 gene (Ecm1): Ecm1a and an alternatively spliced Ecm1b. We studied Ecm1 mRNA expression and localization during endochondral bone formation and investigated the effect of recombinant human (rh) Ecm1a protein on organ cultures of embryonic mouse metatarsals. Of the two transcripts, Ecm1a mRNA was predominantly expressed in fetal metacarpals from day 16 to 19 after gestation. Ecm1 expression was not found in 16- and 17-day-old metatarsals of which the perichondrium was removed. In situ hybridization and immunohistochemistry demonstrated Ecm1 expression in the connective tissues surrounding the developing bones, but not in the cartilage. Biological effects of rhEcm1a protein on fetal metatarsal cultures were biphasic: at low concentrations, Ecm1a stimulated alkaline phosphatase activity and had no effect on mineralization, whereas at higher concentrations, Ecm1a dose dependently inhibited alkaline phosphatase activity and mineralization. These results suggest that Ecm1a acts as a novel negative regulator of endochondral bone formation.

Alkaline Phosphatase↗

Changes in the profile of genistein, daidzein, and their conjugates during thermal processing of tofu.

Profiles of genistein, daidzein, genistin, daidzin, and their acetyl- and malonyl-beta-glycosides were determined in tofu as affected by temperature and time. Tofu was heated in water at 80, 90, and 100 degrees C for 0 (control), 10, 20, 30, and 40 min, and the contents of the isoflavones of interest were quantified using reversed-phase HPLC. Total isoflavone content decreased most likely due to leaching of isoflavones into the water. Because the content of the isoflavones of the genistein series was little affected by the treatments, the decrease in the total isoflavone content was almost exclusively due to a decrease of the daidzein series. Changes in the profile of the daidzein series suggest little decarboxylation of the malonylglycoside to the acetylglycoside, but considerable de-esterification of the malonyl- and acetylglycoside to the beta-glucoside. Strongly temperature dependent decreases of the aglycon suggest possible thermal degradation of daidzein in addition to losses due to leaching.

Chromatography, High Pressure Liquid↗