Biomedical subjects

D Li

Publications and source records attributed to D Li.

At least 181 records · Page 10Linked to original sources

PubMed2001

Tensile bond strengths of fourth- and fifth-generation dentin adhesives with packable resin composites.

PURPOSE: Placement of packable composites would be simplified if a single-bottle dentin bonding agent, rather than the more complex, multiple-bottle dentin bonding agents, involving multiple steps, could be used successfully with adequate bond strength. This study was designed to compare the tensile bond strengths to extracted human dentin of four single-bottle (fifth-generation) and four multiple-bottle (fourth-generation) dentin bonding agents with four packable composites. MATERIALS AND METHODS: Packable composites (P60, 3M Dental Products Division; ALERT, Jeneric-Pentron; Pyramid Dentin, Bisco; and Clearfil AP-X, Kuraray America) were tested for in vitro tensile bond strength to human dentin with their single-bottle and their multiple-bottle dentin bonding agents. Bond strengths were measured using a truncated cone tensile test, with a bonding diameter of 3 mm, after storage of specimens in water at 37 degrees C and 100% relative humidity for 24 hours, and using a testing machine at a crosshead speed of 0.5 mm per minute. RESULTS: Means and standard deviations (n = 10) of tensile bond strengths (MPa) for the single-bottle system were P60, 21.3 MPa (6.7); ALERT, 26.6 MPa (4.2); Pyramid Dentin, 14.8 MPa (4.4); and Clearfil AP-X, 18.6 MPa (3.7). For the multiple-bottle system they were P60, 20.0 MPa (8.6); ALERT, 20.6 MPa (6.2); Pyramid Dentin, 16.0 MPa (3.4); and Clearfil AP-X, 17.8 MPa (6.0). Two-way analysis of variance showed significant differences among composites but no significant differences between single-bottle and multiple-bottle bonding agents. The Tukey-Kramer interval (p < .05) for comparisons of bond strengths among composites was 4.8 MPa. CONCLUSIONS: For single-bottle agents, bond strengths were ranked as ALERT > P60, P60 = Clearfil, P60 > Pyramid, and Clearfil = Pyramid. For multiple-bottle agents, there were no significant differences in bond strengths among the composites. CLINICAL SIGNIFICANCE: Single-bottle bonding agents provide similar in vitro bond strengths for use with packable composites as do the same manufacturers' multiple-bottle bonding agents.

Analysis of Variance↗

PubMed2001

A two-stage microwave image reconstruction procedure for improved internal feature extraction.

We have developed a two-stage Gauss-Newton reconstruction process with an automatic procedure for determining the regularization parameter. The combination is utilized by our microwave imaging system and has facilitated recovery of quantitatively improved images. The first stage employs a Levenberg-Marquardt regularization along with a spatial filtering technique for a few iterations to produce an intermediate image. In effect, the first set of iterative image reconstruction steps synthesizes a priori information from the measurement data versus actually requiring physical prior information on the interrogated object. Because of the interaction of the Levenberg-Marquardt regularization and spatial filtering at each iteration, the intermediate image produced from the first reconstruction stage represents an improvement in terms of the least squared error over the initial uniform guess; however, it has not completely converged in a least squared sense. The second stage involves using this distribution as a priori information in an iteratively regularized Gauss-Newton reconstruction with a weighted Euclidean distance penalty term. The penalized term restricts the final image to a vicinity (determined by the scale of the weighting parameter) about the intermediate image while allowing more flexibility in extracting internal object structures. The second stage makes use of an empirical Bayesian/random effects model that enables an optimal determination of the weighting parameter of the penalized term. The new approach demonstrates quantifiably improved images in simulation, phantom and in vivo experiments with particularly striking improvements with respect to the recovery of heterogeneities internal to large, high contrast scatterers such as encountered when imaging the human breast in a water-coupled configuration.

Algorithms↗

PubMed2001

Modeling of bottom backscattering from three-dimensional volume inhomogeneities and comparisons with experimental data.

In this paper, backscattering from 3D volume inhomogeneities in the seabed is modeled and the results compared with experimental data at 250-650 Hz. The experiment was part of the Acoustic Reverberation Special Research Program (ARSRP) and the data were obtained in a sediment pond on the western flank of the Mid-Atlantic Ridge. A volume scattering model based on first-order perturbation theory is developed incorporating contributions from both sound speed and density fluctuations. With the propagators, i.e., the Green's functions, handled accurately through numerical wave number integration and random fluctuations generated effectively by a new scheme modified from the spectral method, the model is capable of simulating monostatic, backscattered fields in the frequency domain as well as in the time domain owing to 3D volumetric sediment inhomogeneities. The model compares favorably and consistently with the ARSRP backscattering data over the entire frequency band, with the fluctuations of sound speed and density in two irregular sediment layers, identified from the data analysis, described by a power-law type of power spectrum.

Journal Article↗

PubMed2001

Pharmacodynamic assessment of gatifloxacin against Streptococcus pneumoniae.

The pharmacodynamic parameters of peak serum drug concentration/MIC (peak/MIC) ratio and the area under the curve (AUC)/MIC ratio have been used to characterize in vivo drug exposure and its relationship to bacterial killing for the fluoroquinolones. Our study objectives were to describe the pharmacodynamic relationship between gatifloxacin exposure and outcome as assessed by bacterial density and survival in an immunocompromised murine thigh model of pneumococcal infection and to assess the relationship between drug exposure and these outcomes in an immunocompetent host. ICR mice were rendered neutropenic, and thigh infection was induced by intramuscular administration of 0.1 ml of 10(5) to 10(7) CFU of Streptococcus pneumoniae/ml. Mice received 1 to 5 mg of uranyl nitrate/kg of body weight at day -3 and were randomized to receive 10 to 80 mg of gatifloxacin/kg every 6 to 24 h orally, starting at 2 h postinoculation. Bacterial density studies were completed 24 h after initiation of therapy, and survival was assessed after 4 days of treatment. MICs for clinical isolates (n = 8) ranged from 0.25 to 1.0 microg/ml. Correlations were assessed between the change in bacterial density, as well as survival, and the AUC/MIC ratio, peak/MIC ratio, and the duration of time that serum drug concentration remained above the MIC. The best predictor of bacterial response was the AUC/MIC ratio for both outcome measures. There was greater efficacy, as measured by a decrease in log change in CFU as well as by survival data, in the immunocompetent mice compared to the immunocompromised mice. These data demonstrate (i) the appropriateness of the AUC/MIC ratio as a dynamic predictor of response to pneumococcal infection for the fluoroquinolones, (ii) that gatifloxacin AUC/MIC ratios of 30 to 40 appear to optimize bactericidal activity and survival in this model, and (iii) that immunocompetency of the host plays a role in efficacy.

Animals↗

PubMed2001

Domain structure of the NRIF3 family of coregulators suggests potential dual roles in transcriptional regulation.

The identification of a novel coregulator for nuclear hormone receptors, designated NRIF3, was recently reported (D. Li et al., Mol. Cell. Biol. 19:7191-7202, 1999). Unlike most known coactivators, NRIF3 exhibits a distinct receptor specificity in interacting with and potentiating the activity of only TRs and RXRs but not other examined nuclear receptors. However, the molecular basis underlying such specificity is unclear. In this report, we extended our study of NRIF3-receptor interactions. Our results suggest a bivalent interaction model, where a single NRIF3 molecule utilizes both the C-terminal LXXIL (receptor-interacting domain 1 [RID1]) and the N-terminal LXXLL (RID2) modules to cooperatively interact with TR or RXR (presumably a receptor dimer), with the spacing between RID1 and RID2 playing an important role in influencing the affinity of the interactions. During the course of these studies, we also uncovered an NRIF3-NRIF3 interaction domain. Deletion and mutagenesis analyses mapped the dimerization domain to a region in the middle of NRIF3 (residues 84 to 112), which is predicted to form a coiled-coil structure and contains a putative leucine zipper-like motif. By using Gal4 fusion constructs, we identified an autonomous transactivation domain (AD1) at the C terminus of NRIF3. Somewhat surprisingly, full-length NRIF3 fused to the DNA-binding domain of Gal4 was found to repress transcription of a Gal4 reporter. Further analyses mapped a novel repression domain (RepD1) to a small region at the N-terminal portion of NRIF3 (residues 20 to 50). The NRIF3 gene encodes at least two additional isoforms due to alternative splicing. These two isoforms contain the same RepD1 region as NRIF3. Consistent with this, Gal4 fusions of these two isoforms were also found to repress transcription. Cotransfection of NRIF3 or its two isoforms did not relieve the transrepression function mediated by their corresponding Gal4 fusion proteins, suggesting that the repression involves a mechanism(s) other than the recruitment of a titratable corepressor. Interestingly, a single amino acid residue change of a potential phosphorylation site in RepD1 (Ser(28) to Ala) abolishes its transrepression function, suggesting that the coregulatory property of NRIF3 (or its isoforms) might be subjected to regulation by cellular signaling. Taken together, our results identify NRIF3 as an interesting coregulator that possesses both transactivation and transrepression domains and/or functions. Collectively, the NRIF3 family of coregulators (which includes NRIF3 and its other isoforms) may play dual roles in mediating both positive and negative regulatory effects on gene expression.

Alternative Splicing↗

PubMed2001

Contrast-enhanced MR imaging of coronary arteries: comparison of intra- and extravascular contrast agents in swine.

PURPOSE: To compare the efficacy of an intravascular contrast agent, gadomer-17, in improving magnetic resonance (MR) imaging of coronary arteries with that of an extravascular agent, gadopentetate dimeglumine, in pigs. MATERIALS AND METHODS: Eight pigs underwent imaging after three injections: 0.20 mmol of gadopentetate dimeglumine per kilogram of body weight and 0.05 and 0.10 mmol/kg gadomer-17. Coronary images were acquired repeatedly after each injection by using an inversion-recovery-prepared segmented three-dimensional sequence with either breath holding (n = 4) or respiratory gating (n = 4). Coronary artery-to-myocardium contrast-to-noise ratios (CNRs) were compared between injections. RESULTS: At breath-hold imaging, substantial CNR improvement over precontrast images was observed in images acquired during the first pass of gadopentetate dimeglumine in coronary arteries and up to 6 and 10 minutes after 0.05 and 0.10 mmol/kg of gadomer-17 injections, respectively. The CNR with 0.10 mmol/kg of gadomer-17 was 20% (P <.05) higher than that with gadopentetate dimeglumine at first-pass imaging. At respiratory-gated imaging, significant CNR improvement (P <.05) over precontrast images was observed in images acquired up to 10, 30, and 50 minutes after gadopentetate dimeglumine and both gadomer-17 injections, respectively. The CNR on the first images obtained after 0.10 mmol/kg gadomer-17 injection was 168% (P <.05) higher than that on the images obtained after gadopentetate dimeglumine injection. CONCLUSION: Gadomer-17 provided greater and more persistent CNR improvements than did gadopentetate dimeglumine; further evaluation of its utility for coronary imaging in humans is warranted.

Animals↗

PubMed2001

Digital optical imaging of green fluorescent proteins for tracking vascular gene expression: feasibility study in rabbit and human cell models.

PURPOSE: To investigate the feasibility of using a sensitive digital optical imaging technique to detect green fluorescent protein (GFP) expressed in rabbit vasculature and human arterial smooth muscle cells. MATERIALS AND METHODS: A GFP plasmid was transfected into human arterial smooth muscle cells to obtain a GFP-smooth muscle cell solution. This solution was imaged in cell phantoms by using a prototype digital optical imaging system. For in vivo validation, a GFP-lentivirus vector was transfected during surgery into the carotid arteries of two rabbits, and GFP-targeted vessels were harvested for digital optical imaging ex vivo. RESULTS: Optical imaging of cell phantoms resulted in a spatial resolution of 25 microm/pixel. Fluorescent signals were detected as diffusely distributed bright spots. At ex vivo optical imaging of arterial tissues, the average fluorescent signal was significantly higher (P <.05) in GFP-targeted tissues (mean +/- SD, 9,357.3 absolute units of density +/- 1,001.3) than in control tissues (5,633.7 absolute units of density +/- 985.2). Both fluorescence microscopic and immunohistochemical findings confirmed these differences between GFP-targeted and control vessels. CONCLUSION: The digital optical imaging system was sensitive to GFPs and may potentially provide an in vivo imaging tool to monitor and track vascular gene transfer and expression in experimental investigations.

Animals↗

PubMed2001

Coronary arteries: magnetization-prepared contrast-enhanced three-dimensional volume-targeted breath-hold MR angiography.

A volume-targeted contrast agent-enhanced breath-hold coronary magnetic resonance angiographic technique was optimized and evaluated in 16 volunteers. Substantial increases in coronary signal-to-noise ratio, contrast-to-noise ratio, lengths of depiction, and vessel sharpness were observed on enhanced images. The imaging approach with two 20-mL injections of contrast agent covers the left and right coronary arteries in two breath holds and is a promising method for coronary imaging.

Adult↗

PubMed2001

Cine MR angiography of the heart with segmented true fast imaging with steady-state precession.

In five healthy subjects and 18 patients, cine magnetic resonance (MR) imaging of the heart was performed with a true fast imaging with steady-state precession (FISP) sequence. Results were compared both quantitatively and qualitatively with those at cine fast low-angle shot (FLASH) MR imaging. The blood-myocardial contrast-to-noise ratio (CNR) was 2.0 times higher and the normalized (for measurement time and pixel size) blood-myocardial CNR was 4.0 times higher for true FISP compared with FLASH MR imaging. Qualitative scores for image quality were significantly higher with true FISP MR imaging. Segmented cine true FISP MR imaging generated high-contrast MR images of the heart in healthy subjects and in patients with heart disease and produced image quality superior to that with cine FLASH MR imaging.

Adult↗

PubMed2001

Differential gene expression in a murine model of cancer cachexia.

Murine adenocarcinoma 16 (MAC16) tumors and cell lines induce cachexia in NMRI nude mice, whereas histologically similar MAC13 tumors do not. After confirming these findings in BALB/c nude mice, we demonstrated that this tissue wasting was not related to decreased food intake or increased total body oxidative metabolism. Previous studies have suggested that MAC16's cachexigenic properties may involve the production of tumor-specific factors. We therefore screened for genes having increased expression in the MAC16 compared with the MAC13 cell line by performing hybridization to a murine cDNA expression array, by generation and comparison of cDNA libraries from each cell line, and by PCR-based subtractive hybridization. Northern blot hybridization was performed to confirm differences in transcript expression. Transcripts encoding insulin-like growth factor binding protein-4, cathepsin B, ferritin light and heavy chain, endogenous long-terminal repeat sequences, and a viral envelope glycoprotein demonstrated increased expression in the MAC16 cell line. The roles of a number of these genes in known metabolic pathways identify them as potential participants in the induction of cachexia.

Adenocarcinoma↗

PubMed2001

TGF-beta(1) modulates NOS expression and phosphorylation of Akt/PKB in rat myocytes exposed to hypoxia-reoxygenation.

Myocardial hypoxia-reoxygenation (H-R) is associated with upregulation of inducible nitric oxide synthase (iNOS), decrease in endothelial NOS (eNOS), and increase in protein kinase B (PKB). Previous work also shows that transforming growth factor-beta(1) (TGF-beta(1)) can attenuate myocardial injury induced by H-R. We examined the modulation of NOS and PKB expression in response to H-R by TGF- beta(1). Myocytes from Sprague-Dawley rat hearts were cultured and exposed to hypoxia (95% N(2)-5% CO(2), PO(2) ~30 mmHg) for 24 h and reoxygenation (95% air-5% CO(2)) for 3 h. Myocytes were then examined for lactate dehydrogenase (LDH) release, iNOS activity (conversion of L-[(3)H]arginine to L-[(3)H]citrulline), iNOS and eNOS expression, and PKB phosphorylation. H-R alone resulted in myocyte injury, upregulation of iNOS activity and expression, decrease in eNOS expression, and increase in PKB phosphorylation (all P < 0.05 vs. cells cultured in normoxic conditions). Treatment of myocytes with TGF-beta(1) (1 ng/ml) resulted in a reduction in LDH release, attenuation of the alterations in NOS expression (both iNOS and eNOS), and PKB phosphorylation in response to H-R (all P < 0.05 vs. H-R alone). These observations suggest that TGF-beta(1) decreases H-R injury and attenuates alterations in NOS and PKB phosphorylation in myocytes exposed to H-R.

Animals↗

PubMed2001

Upregulation of lung soluble guanylate cyclase during chronic hypoxia is prevented by deletion of eNOS.

Hypoxia upregulates endothelial (e) nitric oxide synthase (NOS), but how eNOS affects soluble guanylate cyclase (sGC) protein expression in hypoxia-induced pulmonary hypertension is unknown. Wild-type (WT), eNOS-deficient [eNOS(-/-)], and inducible NOS (iNOS)-deficient [iNOS(-/-)] mice were used to investigate the effects of lack of NO from different NOS isoforms on sGC activity and protein expression and its relationship to the muscularization of the pulmonary vasculature. After 6 days of hypoxic exposure (10% O2), the ratios of the right ventricle to left ventricle + septum weight (RV/LV+S) and right ventricle weight to body weight, the lung sGC activity, and vascular muscularization were determined, and protein analysis for eNOS, iNOS, and sGC was performed. Results demonstrated that there were significant increases of RV/LV+S in all animals treated with hypoxia. In hypoxic WT and iNOS(-/-) mice, eNOS and sGC alpha1- and beta1-protein increased twofold; cGMP levels and the number of muscularized vessels also increased compared with hypoxic eNOS(-/-) mice. There was a twofold increase of iNOS protein in WT and eNOS(-/-) mice, and the basal iNOS protein concentration was higher in eNOS(-/-) mice than in WT mice. In contrast, the eNOS(-/-) mouse lung showed no eNOS protein expression, lower cGMP concentrations, and no change of sGC protein levels after hypoxic exposure compared with its normoxic controls (P > 0.34). These results suggest that eNOS, but not iNOS, is a major regulator of sGC activity and protein expression in the pulmonary vasculature.

Animals↗

PubMed2001

Angiotensin II via activation of type 1 receptor upregulates expression of endoglin in human coronary artery endothelial cells.

Transforming growth factor-beta1 and its subtype receptor endoglin are key components in angiogenesis. We explored the role of angiotensin (Ang) II in the expression of endoglin and the underlying intracellular signaling mechanism in human coronary artery endothelial cells. Incubation of cells with Ang II upregulated endoglin expression in a concentration- and time-dependent manner (maximal effect with 10(-6) mol/L Ang II at 24 hours). The Ang II type 1 receptor blocker losartan, but not the type 2 receptor blocker PD 123,319, completely blocked the effect of Ang II. In parallel experiments, the mitogen-activated protein kinase inhibitor PD 098,059 fully inhibited the effect of Ang II on the expression of endoglin. Incubation of endothelial cells with Ang II also increased the expression of transforming growth factor-beta1 and -beta2 receptors and simultaneously decreased the levels of transforming growth factor-beta1. These effects of Ang II were also attenuated by losartan. We propose that Ang II via its type 1 receptor activation modulates the expression of transforming growth factor-beta1 receptors in human coronary endothelial cells. The activation of mitogen-activated protein kinase plays an important role in this process. These observations provide a new clue regarding the regulatory effect of Ang II on vascular remodeling after injury.

Angiotensin II↗

PubMed2001

Different isoforms of tocopherols enhance nitric oxide synthase phosphorylation and inhibit human platelet aggregation and lipid peroxidation: implications in therapy with vitamin E.

BACKGROUND: alpha-Tocopherol has received much attention in the primary and secondary prevention of coronary artery disease. Absence of other isoforms, such as gamma- and delta-tocopherol, in commercial preparations of vitamin E may account for the inconsistent results of clinical trials. Since platelet aggregation is intimately involved in thrombogenesis, the relative effects of alpha-, gamma-, and delta-tocopherol and their combination were examined on human platelet aggregation, lipid peroxidation, and constitutive nitric oxide synthase (cNOS) activity. METHODS AND RESULTS: Human platelets were incubated with the three different isoforms of tocopherol and their combination for 30 minutes, and then ADP-induced platelet aggregation measured. All three isoforms of tocopherol markedly and similarly decreased platelet aggregation in a concentration (120--480 microM)-dependent manner. All three tocopherols also decreased the level of the lipid peroxidation product, malondialdehyde (MDA), and increased NO release (P < 0.05 vs control). These isoforms of tocopherol did not affect cNOS protein expression, but enhanced cNOS phosphorylation in platelets. The combination of three tocopherols in a concentration found in nature was more potent than alpha-, gamma-, or delta-tocopherol alone in this regard. CONCLUSION: These observations suggest that all three major isoforms of tocopherol have a similar effect on human platelet aggregation. The three isoforms appear to attenuate platelet aggregation at least in part via a decrease in free radical generation and an increase in platelet cNOS activity. The combination of tocopherols has a synergistic platelet inhibitory effect. Future clinical trials should concentrate on the combination of these three isoforms of tocopherols.

Analysis of Variance↗

PubMed2001

Distribution and regulation of galanin-like peptide (GALP) in the hypothalamus of the mouse.

Galanin-like peptide (GALP) is a newly discovered molecule whose expression in the brain is confined to the arcuate nucleus and median eminence. In the rat, cellular levels of GALP mRNA are reduced by fasting and reversed by peripheral administration of leptin. The purpose of this investigation was 1) to clone and map the distribution of GALP mRNA in the brain of the mouse; 2) to compare the pattern and magnitude of GALP mRNA expression in the leptin-deficient obese (ob/ob) mouse with that of wild-type controls; and 3) to examine the effects of leptin delivered into the brain on the expression of GALP mRNA in the ob/ob mouse. We report the sequence of a mouse GALP cDNA and show that GALP mRNA is expressed in the arcuate nucleus, median eminence, infundibular stalk, and the neurohypophysis of this species. The expression of GALP mRNA in the brain was markedly reduced in the ob/ob mice, compared with wild-type animals. Intracerebroventricular infusion of leptin to ob/ob mice increased both the number of GALP mRNA-expressing neurons and their content of GALP mRNA, compared with vehicle-treated controls. These observations demonstrate that GALP mRNA is induced by leptin through a direct action on the brain.

Animals↗

PubMed2001

Inhibition of heat shock transcription factor by GR.

The GR is a hormone-activated transcription factor that acts to regulate specific gene expression. In the absence of hormone, the GR and other steroid receptors have been shown to form complexes with several mammalian heat shock proteins. As heat shock proteins are produced by cells as an adaptive response to stress, speculation has existed that communication between the heat shock and glucocorticoid hormone signal pathways must exist. Only recently has evidence to support this hypothesis been reported. In almost all cases, the evidence has been of an ability of heat shock to cause a potentiation of the glucocorticoid hormone response. In this proposal, evidence is now presented that heat shock signaling can, in turn, be regulated by glucocorticoids. In mouse L929 cells stably expressing a chloramphenicol acetyltransferase reporter controlled by the human heat shock protein70 promoter and containing known binding sites for heat shock transcription factor 1 treatment with glucocorticoid agonist (dexamethasone) results in a dose-dependent decrease of stress-induced chloramphenicol acetyltransferase gene expression. In these cells, inhibition of heat shock protein70 promoter activity by dexamethasone was completely blocked by GR antagonist (RU486). Similar treatment of L929 cells stably expressing a chloramphenicol acetyltransferase reporter under the control of the constitutively active SV40 promoter showed no such inhibition by dexamethasone. More importantly, dexamethasone was also found to inhibit heat shock-induced expression of the major heat shock proteins-heat shock proteins70, 90, and 110. Thus, the inhibitory effect of dexamethasone appears to apply to most, if not all, heat shock transcription factor 1-regulated genes. Although dexamethasone did not prevent the DNA-binding function of heat shock-activated heat shock transcription factor 1, it did inhibit a constitutively active mutant of human heat shock transcription factor 1 under nonstress conditions, suggesting that dexamethasone repression of heat shock transcription factor 1 was primarily through an inhibition of heat shock transcription factor 1 transcription enhancement activity. To more accurately characterize the stage of GR signaling responsible for inhibition of heat shock transcription factor 1, a series of Chinese hamster ovary cells containing either no GR, wild-type mouse GR, or single-point mutations of GR were employed. Dexamethasone inhibition of heat shock-induced heat shock transcription factor 1 activity was observed in the presence of wild-type GR, but not in Chinese hamster ovary cells lacking GR, suggesting that signaling cascades other than GR were not involved in this effect of dexamethasone. Consistent with this conclusion was the observation that dexamethasone had no effect on activity of the MAPKs (ERK1, ERK2, or c-jun N-terminal kinase), which are known to negatively regulate heat shock transcription factor 1. Dexamethasone inhibition of heat shock transcription factor 1 was not seen in Chinese hamster ovary cells expressing GR defective for DNA-binding function. Moreover, dissociation of GR/Hsp90/Hsp70 complexes was observed in response to hormone for both the wild-type and DNA binding-defective forms of GR, demonstrating that release of Hsp90 or Hsp70 (both of which are known to keep heat shock transcription factor 1 in its inactive state) could be ruled out as a potential mechanism. Thus, it appears that GR-mediated transactivation or transrepression is required for the inhibitory effect of dexamethasone on heat shock transcription factor 1 activity. Taken as a whole, these results provide evidence for a novel mechanism of cross-talk in which signaling by the GR can attenuate the heat shock response in cells through an inhibition of the transcription enhancement activity of HSF1.

Animals↗

PubMed2001

Formamidopyrimidine-DNA glycosylase enhances arsenic-induced DNA strand breaks in PHA-stimulated and unstimulated human lymphocytes.

To confirm that arsenic (As) induces oxidative DNA damage in phytohemagglutinin (PHA)-stimulated and unstimulated human lymphocytes, we used the alkaline comet assay combined with specific enzyme [formamidopyrimidine-DNA glycosylase (FPG)] digestion to measure As-induced base damage. The results showed that the enzyme-sensitive sites were readily detected with the alkaline comet assay after the cells were treated with 10 microM As for 2 hr. The repair patterns observed for FPG-created DNA single strand breaks (SSBs) in As-treated cells were comparable to those in hydrogen peroxide (H(2)O(2))-treated cells. The enzyme-created SSBs, As-induced base damage, were more significant in PHA-stimulated lymphocytes. About 63% and 68% of SSBs induced by As and H(2)O(2), respectively, were repaired in PHA-stimulated lymphocytes by 2-hr repair incubation, but about 34% and 43%, respectively, were repaired in unstimulated cells. About 40% and 49% of base damage induced by As and H(2)O(2), respectively, were repaired in PHA-stimulated lymphocytes, but about 19% and 21%, respectively, were repaired in unstimulated cells. These results indicated that As induced oxidative DNA damage in human lymphocytes at micromolar concentrations. The damaged bases could be chiefly purines or formamidopyrimidines. Like the damage induced by H(2)O(2), As-induced DNA damage was repaired more slowly in unstimulated lymphocytes.

Arsenic↗

PubMed2001

Developmental changes in galanin in lumbosacral sympathetic ganglionic neurons innervating the avian uterine oviduct and galanin induction by sex steroids.

We recently found lumbosacral sympathetic ganglionic galanin neurons innervating the quail uterine oviduct. Galaninergic innervation of the uterine muscle may be essential for avian oviposition, as galanin evoked oviposition through a mechanism of induction of vigorous uterine contraction. The questions arising from these findings are: what changes occur in galanin expression in the sympathetic ganglionic galanin neuron during development, and what is the hormonal factor(s) that induces galanin expression in this neuron? Therefore, the present study examined the developmental changes in galanin of the quail sympathetic ganglionic neuron and uterus, and the effect of administration of ovarian sex steroids on galanin induction. Immature birds reared under long-day photoperiods from 4 weeks of age demonstrated progressive increases in galanin levels both per unit ganglionic protein (concentration) and per ganglia (content) concurrent with ganglionic development during weeks 4--13. The uterine galanin content and uterine weight also increased progressively during the same period, but the galanin concentration in the uterus at 4 weeks was high due to the much smaller tissue mass. Immunocytochemical analysis with anti-galanin serum showed that immunoreactive ganglionic cells were few and small at 4 weeks and increased progressively thereafter. Administration of oestradiol-17 beta to immature birds at 3 weeks of age for 1 week increased both the galanin concentration and content in the ganglia without ganglionic growth. A marked increase in galanin-immunoreactive ganglionic cells was detected following oestradiol treatment. In contrast, progesterone increased ganglionic galanin levels, but the effects were low. Expression of the mRNAs encoding oestrogen receptor-alpha and -beta (ER alpha and ER beta) in the ganglionic tissue was verified by RT-PCR/Southern blot analysis. Immunocytochemical staining with anti-ER serum further revealed an intense immunoreaction restricted to the nucleus of ganglionic neurons. These results suggest that ovarian sex steroids, in particular oestradiol-17 beta, contribute as hormonal factors to galanin induction, which takes place in the lumbosacral sympathetic ganglionic neurons innervating avian uterine oviduct during development. Oestradiol may act directly on this ganglionic neuron through intra-nuclear receptor-mediated mechanisms to induce galanin.

Animals↗