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In Vivo (in + vivo)
Terms modified by In Vivo Selected AbstractsG,q -PROTEIN CARBOXYL TERMINUS IMITATION POLYPEPTIDE GCIP-27 ATTENUATES CARDIAC HYPERTROPHY IN VITRO AND IN VIVOCLINICAL AND EXPERIMENTAL PHARMACOLOGY AND PHYSIOLOGY, Issue 12 2007Hai-Gang Zhang SUMMARY 1Various Gq -protein-coupled receptors, such as ,1 -adrenoceptors, angiotension AT1 receptors, endothelin ETA receptors, neuropeptide Y1 receptors etc., contribute to cardiac hypertrophy. In G-protein signalling pathways, the carboxyl terminus of the G, subunit plays a vital role within G-protein,receptor interaction. The present study was designed to explore the effects of the synthetic G,q carboxyl terminal imitation peptide GCIP-27 on cardiac hypertrophy. 2Hypertrophy of rat cultured cardiomyocytes was induced by noradrenaline (NA) or angiotensin (Ang) II in vitro. Protein content, [3H] incorporation and [Ca2+]i were determined in cardiomyocytes cultured with GCIP-27. Three in vivo animal models of cardiac hypertrophy were prepared using intraperitoneal injections of NA in mice and rats and suprarenal abdominal aortic stenosis in rats. After treatment with GCIP-27 (10,100 µg/L) for 15 or 20 days, indices of cardiac hypertrophy were measured. The effect of GCIP-27 on the mRNA expression of c-fos and c-jun was detected using reverse transcription,polymerase chain reaction. 3At 10,100 µg/L, GCIP-27 significantly decreased protein content and [3H]-leucine incorporation in cultured cardiomyocytes compared with 1 µmol/L NA- and 1 µmol/L AngII-treated groups. After treatment with GCIP-27 (10, 30 or 100 µg/kg) for 15 days, the heart index (HI) and left ventricular index (LVI) in mice decreased significantly compared with the NA control group. In rats, GCIP-27 significantly reduced HI and LVI compared with the NA and aortic stenosis groups. Moreover, [Ca2+]i in cardiomyocytes in the GCIP-27 (3, 10, 30 µg/L)-treated groups was lower than that in the control groups. Expression of c-fos and c-jun mRNA decreased significantly in the myocardium from 5,45 µg/L GCIP-27-treated rats compared with NA controls. 4The results indicate that GCIP-27 can attenuate cardiac hypertrophy effectively in various models in vitro and in vivo. [source] Fibronectin Functionalized Hydroxyapatite Coatings: Improving Dermal Fibroblast Adhesion In Vitro and In Vivo,ADVANCED ENGINEERING MATERIALS, Issue 8 2010Catherine J. Pendegrass Skin-penetrating devices including intraosseous transcutaneous amputation prostheses (ITAP) and external fixator pins rely on a skin-implant seal to prevent infection. In this study, we assess the effectiveness of fibronectin (Fn) functionalized hydroxyapatite (HA) coatings for promoting dermal fibroblast and dermal tissue attachment and ingrowth in vitro and in vivo. By measuring the number of focal adhesions per unit cell area we have demonstrated that HA significantly promotes dermal fibroblast attachment compared with titanium alloy. Dermal fibroblast attachment is promoted further using Fn functionalized HA coatings incorporated into an implant design with 700,µm pores, which significantly increased dermal tissue ingrowth and attachment compared with non-functionalized HA and titanium alloy controls incorporating 500 or 1000,µm pores. We postulate that Fn functionalized HA coatings applied to transdermal implants may promote and sustain the skin-implant interface and assist in preventing infection long term. [source] Influence of TiO2 Nanoparticles Incorporated into Elastomeric Polyesters on their Biocompatibility In Vitro and In VivoADVANCED ENGINEERING MATERIALS, Issue 11 2009Miroslawa El-Fray Abstract Fibroblasts proliferation and apoptosis as well as tissue response after implantation of elastomers containing nanocrystalline TiO2 were investigated in the present in vitro and in vivo study. Materials investigated were soft poly(aliphatic/aromatic-ester) multiblock thermoplastic elastomers with poly(ethylene terephthalate) (PET) hard segments and dimerized linoleic acid (DLA) soft segments, respectively, containing 0.2,wt% TiO2 nanoparticles. An investigation of the influence of TiO2 nanoparticles incorporated into polymeric material on in vitro biocompatibility revealed enhanced cell proliferation and diminished number of necrotic and apoptotic cells as compared to nanoparticles-free polymer. Implantation tests indicated that the observed tissue changes were similar to those observed with medical-grade silicone elastomer, no evidence of contact necrosis being observed. The unchanged morphology of rat liver hepatocytes and the lack of parenchymal necrosis also indicated that exposure to the material containing TiO2 nanoparticles, did not cause any cytotoxic reactions. The present study, thus, showed that elastomeric polyester containing TiO2 nanoparticles are interesting biomimetic constructs for improved tissue regeneration. [source] The Neurogenic Vasodilator Response to Endothelin-1: A Study in Human Skin In VivoEXPERIMENTAL PHYSIOLOGY, Issue 6 2000Ruwani Katugampola We have investigated the mediators and mechanisms underlying the vasodilator effects of the potent vasoactive peptide, endothelin-1 (ET-1) and its isomers ET-2 and ET-3 in human skin, in vivo, using cutaneous microdialysis to quantify the release of mediators within the dermal response and scanning laser Doppler imaging to measure changes in blood flux. The effects of local anaesthesia, inhibition of nitric oxide synthase (NOS) by L-NAME and ET receptor blockade on the ET-induced vascular response were also investigated. ET-1, -2 and -3 all caused a dose-dependent area of pallor surrounded by a long-lasting flare which was accompanied by a short-lived burning pruritus. The concentration of nitric oxide (NO) in dialysate collected within the pallor response to 5 ,M ET-1 (1.43 ± 0.64 ,M, n = 5) was not significantly different from baseline levels collected prior to injection (0.86 ± 0.38 ,M) whilst that in the flare increased to reach a peak value of 2.28 ± 0.61 ,M at between 4 and 10 min after intradermal injection (P < 0.004). Pretreatment with local anaesthetic slowed the development of the flare and significantly reduced its size by up to 52% at 20 min after injection (P < 0.05) but had no significant effect on the central pallor. L-NAME, delivered by dialysis also caused a significant reduction in the ET-1-induced flare (P < 0.005). Bosentan, the non-selective ETA/ETB antagonist, when given by dialysis at the site of injection, reduced the area of both the ET-1-induced pallor and surrounding flare by 41 and 26%, respectively. No significant increase in tissue histamine was measured within either the pallor or flare response to ET-1, -2 or -3. Together these data confirm that the vasodilator response to endothelin-1 in human skin is neurogenic in origin and that it is in part mediated by the local release of nitric oxide. There appears to be little evidence for the involvement of mast cell-derived histamine in the initiation or modulation of ET-induced vasodilatation, in vivo. [source] Controlled Degradability of Polysaccharide Multilayer Films In Vitro and In Vivo,ADVANCED FUNCTIONAL MATERIALS, Issue 11 2005C. Picart Abstract This article demonstrates the possibility of tuning the degradability of polysaccharide multilayer films in vitro and in vivo. Chitosan and hyaluronan multilayer films (CHI/HA) were either native or crosslinked using a water soluble carbodiimide, 1-ethyl-3-(3-dimethylamino-propyl)carbodiimide (EDC) at various concentrations in combination with N-hydroxysulfosuccinimide. The in-vitro degradation of the films in contact with lysozyme and hyaluronidase was followed by quartz crystal microbalance measurements, fluorimetry, and confocal laser scanning microscopy after labeling of the chitosan with fluorescein isothiocyanate (CHIFITC). The native films were subjected to degradation by these enzymes, and the crosslinked films were more resistant to enzymatic degradation. Films made of chitosan of medium molecular weight were more resistant than films made of chitosan-oligosaccharides. The films were also brought in contact with plasma, which induced a change in film structure for the native film but did not have any effect on the crosslinked film. The in-vitro study shows that macrophages can degrade all types of films and internalize the chitosan. The in-vivo degradation of the films implanted in mouse peritoneal cavity for a week again showed an almost complete degradation of the native films, whereas the crosslinked films were only partially degraded. Taken together, these results suggest that polysaccharide multilayer films are of potential interest for in-vivo applications as biodegradable coatings, and that degradation can be tuned by using chitosan of different molecular weights and by controlling film crosslinking. [source] Metabolism of the mesoionic compound (MI-D) by mouse liver microsome, detection of its metabolite In Vivo, and acute toxicity in miceJOURNAL OF BIOCHEMICAL AND MOLECULAR TOXICOLOGY, Issue 6 2009Silvia Romão Abstract The mesoionic derivative 4-phenyl-5-[4-nitrocinnamoyl]-1,3,4-thiadiazolyl-2-phenylamine chloride (MI-D) has antitumoral and anti-inflammatory effects. In this study, we present aspects of its metabolism and toxicity in mice. MI-D was metabolized in vitro by liver microsome, generating a main product with a much shorter retention time than MI-D in high-performance liquid chromatography (HPLC) analysis but with a spectrum similar to that of the original molecule. Mass spectrometry with electrospray ionization in positive mode analysis of the purified compound by HPLC indicated that the product of metabolism has four additional hydroxyl groups (m/z = 465) compared with MI-D (m/z = 401). The HPLC analyses of plasma and urine samples from mice treated with MI-D showed the presence of the metabolite product. The kinetic parameters Km (19.5 ± 4.5 ,M) and Vmax [1.5 ± 0.4 units of fluorescence/(100 ,g of microsomal protein/mL/s)] were estimated, confirming the metabolism of MI-D and indicating that the reaction follows Michaelis-Menten kinetics. Acute toxicity was established on the basis of an estimation of mean lethal dose (LD-50; 181.2 mg/kg) and histopathological analysis of animals that survived the LD-50 test. Abdominal adhesions, inflammatory foci, and formation of granulomas were observed. Altogether, the results contribute to the advancement of research in support of MI-D as a future chemotherapeutic drug. © 2009 Wiley Periodicals, Inc. J Biochem Mol Toxicol 23:394,405, 2009; Published online in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/jbt.20303 [source] Connexin 43 Is Required for the Anti-Apoptotic Effect of Bisphosphonates on Osteocytes and Osteoblasts In Vivo,JOURNAL OF BONE AND MINERAL RESEARCH, Issue 11 2008Lilian I Plotkin Abstract Connexin (Cx)43 is required for inhibition of osteocyte and osteoblast apoptosis by bisphosphonates in vitro. Herein, we evaluated its requirement for the in vivo actions of bisphosphonates using mice in which Cx43 was deleted specifically from osteocytes and osteoblasts (Cx43,Ob,Ot/, mice). Effective removal of Cx43 was confirmed by the presence of the deleted form of the gene and by reduced mRNA and protein expression in osteoblastic cells and bones obtained from Cx43,Ob,Ot/, mice. The amino-bisphosphonate alendronate (2.3 ,mol/kg/d) was injected daily into 5-mo-old female mice (n = 6,11) for 31 days, starting 3 days before implantation of pellets releasing the glucocorticoid prednisolone (2.1 mg/kg/d). Cx43,Ob,Ot/, mice and their littermates (Cx43fl/,, Cx43,Ob,Ot/+, and Cx43fl/+) gained bone with similar kinetics and exhibited identical bone mass from 2 to 4.5 mo of age, indicating that Cx43 deletion from osteocytes and mature osteoblasts does not impair bone acquisition. In addition, prednisolone induced a similar increase in osteocyte and osteoblast apoptosis in Cx43,Ob,Ot/, or in control Cx43fl/, littermates. However, whereas alendronate prevented prednisolone-induced apoptosis in control Cx43fl/, mice, it was ineffective in Cx43,Ob,Ot/, mice. In contrast, alendronate inhibited glucocorticoid-induced bone loss in both type of animals, suggesting that inhibition of resorption is the predominant effect of alendronate against the early phase of glucocorticoid-induced bone loss. Taken together with earlier in vitro evidence, these findings show that Cx43 is required for the anti-apoptotic effect of bisphosphonates on osteocytes and osteoblasts. [source] Monitoring Teriparatide-Associated Changes in Vertebral Microstructure by High-Resolution CT In Vivo: Results From the EUROFORS Study,JOURNAL OF BONE AND MINERAL RESEARCH, Issue 9 2007Christian Graeff Dipl-Ing Abstract We introduce a method for microstructural analysis of vertebral trabecular bone in vivo based on HRCT. When applied to monitor teriparatide treatment, changes in structural variables exceeded and were partially independent of changes in volumetric BMD. Introduction: Monitoring of osteoporosis therapy based solely on bone densitometry is insufficient to assess anti-fracture efficacy. Assessing bone microstructure in vivo is therefore of importance. We studied whether it is possible to monitor effects of teriparatide on vertebral trabecular microstructure independent of BMD by high-resolution CT (HRCT). Materials and Methods: In a subset of 65 postmenopausal women with established osteoporosis who participated in the EUROFORS study, HRCT scans of T12, quantitative CT of L1,L3, and DXA of L1,L4 were performed after 0, 6, and 12 mo of teriparatide treatment (20 ,g/d). We compared BMD and 3D microstructural variables in three groups of women, based on prior antiresorptive treatment: treatment-naïve; pretreated; and pretreated women showing inadequate response to treatment. Results: We found statistically highly significant increases in most microstructural variables and BMD 6 mo after starting teriparatide. After 12 mo, apparent bone volume fraction (app. BV/TV) increased by 30.6 ± 4.4% (SE), and apparent trabecular number (app. Tb.N.) increased by 19.0 ± 3.2% compared with 6.4 ± 0.7% for areal and 19.3 ± 2.6% for volumetric BMD. The structural changes were partially independent of BMD as shown by a significantly larger standardized increase and a standardized long-term precision at least as good as DXA. Patients who had shown inadequate response to prior osteoporosis treatment did show improvements in BMD and structural measures comparable to treatment-naïve patients. Conclusions: HRCT is a feasible method for longitudinal microstructural analysis of human vertebrae in vivo, offers information beyond BMD, and is sufficiently precise to show profound effects of teriparatide after 12 mo. [source] Bone Morphogenetic Protein 2 Induces Cyclo-oxygenase 2 in Osteoblasts via a Cbfa1 Binding Site: Role in Effects of Bone Morphogenetic Protein 2 In Vitro and In VivoJOURNAL OF BONE AND MINERAL RESEARCH, Issue 10 2005Daichi Chikazu Abstract We tested the hypothesis that induction of cyclo-oxygenase (COX) 2 mediates some effects of bone morphogenetic protein (BMP) 2 on bone. BMP-2 induced COX-2 mRNA and prostaglandin (PG) production in cultured osteoblasts. BMP-2 increased luciferase activity in calvarial osteoblasts from mice transgenic for a COX-2 promoter-luciferase reporter construct (Pluc) and in MC3T3-E1 cells transfected with Pluc. Deletion analysis identified the -300/-213-bp region of the COX-2 promoter as necessary for BMP-2 stimulation of luciferase activity. Mutation of core-binding factor activity 1 (muCbfa1) consensus sequence (5,-AACCACA-3,) at -267/-261 bp decreased BMP-2 stimulation of luciferase activity by 82%. Binding of nuclear proteins to an oligonucleotide spanning the Cbfa1 site was inhibited or supershifted by specific antibodies to Cbfa1. In cultured osteoblasts from calvariae of COX-2 knockout (-/-) and wild-type (+/+) mice, the absence of COX-2 expression reduced the BMP-2 stimulation of both ALP activity and osteocalcin mRNA expression. In cultured marrow cells flushed from long bones, BMP-2 induced osteoclast formation in cells from COX-2+/+ mice but not in cells from COX-2,/, mice. In vivo, BMP-2 (10 ,g/pellet) induced mineralization in pellets of lyophilized collagen implanted in the flanks of mice. Mineralization of pellets, measured by microcomputed tomography (,CT), was decreased by 78% in COX-2,/, mice compared with COX-2+/+ mice. We conclude that BMP-2 transcriptionally induces COX-2 in osteoblasts via a Cbfa1 binding site and that the BMP-2 induction of COX-2 can contribute to effects of BMP-2 on osteoblastic differentiation and osteoclast formation in vitro and to the BMP-2 stimulation of ectopic bone formation in vivo. [source] Strong Static Magnetic Field Stimulates Bone Formation to a Definite Orientation In Vitro and In Vivo,JOURNAL OF BONE AND MINERAL RESEARCH, Issue 10 2002Hiroko Kotani Ph.D. Abstract The induction of bone formation to an intentional orientation is a potentially viable clinical treatment for bone disorders. Among the many chemical and physical factors, a static magnetic field (SMF) of tesla order can regulate the shapes of blood cells and matrix fibers. This study investigated the effects of a strong SMF (8 T) on bone formation in both in vivo and in vitro systems. After 60 h of exposure to the SMF, cultured mouse osteoblastic MC3T3-E1 cells were transformed to rodlike shapes and were orientated in the direction parallel to the magnetic field. Although this strong SMF exposure did not affect cell proliferation, it up-regulated cell differentiation and matrix synthesis as determined by ALP and alizarin red stainings, respectively. The SMF also stimulated ectopic bone formation in and around subcutaneously implanted bone morphogenetic protein (BMP) 2-containing pellets in mice, in which the orientation of bone formation was parallel to the magnetic field. It is concluded that a strong SMF has the potency not only to stimulate bone formation, but also to regulate its orientation in both in vitro and in vivo models. This is the first study to show the regulation of the orientation of adherent cells by a magnetic field. We propose that the combination of a strong SMF and a potent osteogenic agent such as BMP possibly may lead to an effective treatment of bone fractures and defects. [source] Midregion Parathyroid Hormone-Related Protein Inhibits Growth and Invasion In Vitro and Tumorigenesis In Vivo of Human Breast Cancer CellsJOURNAL OF BONE AND MINERAL RESEARCH, Issue 12 2001Claudio Luparello Abstract Parathyroid hormone-related protein (PTHrP) is critical for normal mammary development and is overexpressed by breast cancers. PTHrP is a peptide hormone that undergoes extensive post-translational processing, and PTHrP(38,94)-amide is one of the mature secretory forms of the peptide. In this study, we explored the effect of PTHrP(38,94)-amide in a panel of six breast cancer cell lines "in vitro" and in MDA-MB231 cells "in vivo" specifically examining cell viability, proliferation, invasiveness, and growth in nude mice. PTHrP(38,94)-amide markedly inhibited proliferation and also caused striking toxicity and accelerated cell death in breast cancer cells. In addition, direct injection of PTHrP(38,94)-amide into MDA-MB231 breast cancer cells passaged in immunodeficient mice produced a marked reduction in tumor growth. These studies (i) indicate breast cancer cells are one of the few tissues in which specific effects of midregion PTHrP have been established to date, (ii) support a role for midregion secretory forms of PTHrP in modulating not only normal but also pathological mammary growth and differentiation, (iii) add further evidence for the existence of a specific midregion PTHrP receptor, and (iv) provide a novel molecule for modeling of small molecule analogues that may have anti-breast cancer effects. [source] Loss of Osteocyte Integrity in Association with Microdamage and Bone Remodeling After Fatigue In Vivo,JOURNAL OF BONE AND MINERAL RESEARCH, Issue 1 2000Olivier Verborgt Abstract As a result of fatigue, bone sustains microdamage, which is then repaired by bone-remodeling processes. How osteoclastic activity is targeted at the removal of microdamaged regions of bone matrix is unknown. In the current studies, we tested the hypothesis that changes in osteocyte integrity, through the initiation of regulated cell death (apoptosis), are associated with fatigue-related microdamage and bone resorption. Ulnae of adult rats were fatigue-loaded to produce a known degree of matrix damage. Osteocyte integrity was then assessed histomorphometrically from terminal deoxynucleotidyl transferase,mediated deoxyuridine triphosphate,nick end labeling (TUNEL),stained sections to detect cells undergoing DNA fragmentation associated with apoptosis; toluidine blue,stained sections were used for secondary morphological confirmation. Ten days after loading, large numbers of TUNEL-positive osteocytes were found in bone surrounding microcracks and in bone surrounding intracortical resorption spaces (,300% increases over controls, p < 0.005). TUNEL labeling in loaded ulnae at sites distant from microcracks or resorption foci did not differ from that in control bone. Osteocytes in toluidine blue,stained sections showed equivalent trends to TUNEL-stained sections, with significant increases in pyknotic nuclei and empty lacunae associated with microcracks and intracortical resorption spaces. TUNEL-positive osteocytes were observed around bone microdamage by 1 day after loading (p < 0.01 relative to baseline), and their number remained elevated throughout the entire experimental period. Increases in empty lacunae and decreases in normal osteocyte numbers were observed over time as well. These studies show that (1) osteocyte apoptosis is induced by bone fatigue, (2) this apoptosis is localized to regions of bone that contain microcracks, and (3) osteoclastic resorption after fatigue also coincides with regions of osteocyte apoptosis. The strong associations between microdamage, osteocyte apoptosis, and subsequent bone remodeling support the hypothesis that osteocyte apoptosis provides a key part of the activation or signaling mechanisms by which osteoclasts target bone for removal after fatigue-induced matrix injury. [source] Role of the Latent Transforming Growth Factor ,,Binding Protein 1 in Fibrillin-Containing Microfibrils in Bone Cells In Vitro and In VivoJOURNAL OF BONE AND MINERAL RESEARCH, Issue 1 2000Sarah L. Dallas Abstract Latent transforming growth factor ,,binding proteins (LTBPs) are extracellular matrix (ECM) proteins that bind latent transforming growth factor , (TGF-,) and influence its availability in bone and other connective tissues. LTBPs have homology with fibrillins and may have related functions as microfibrillar proteins. However, at present little is known about their structural arrangement in the ECM. By using antibodies against purified LTBP1, against a short peptide in LTBP1, and against epitope-tagged LTBP1 constructs, we have shown colocalization of LTBP1 and fibrillin 1 in microfibrillar structures in the ECM of cultured primary osteoblasts. Immunoelectron microscopy confirmed localization of LTBP1 to 10- to 12-nm microfibrils and suggested an ordered aggregation of LTBP1 into these structures. Early colocalization of LTBP1 with fibronectin suggested a role for fibronectin in the initial assembly of LTBP1 into the matrix; however, in more differentiated osteoblast cultures, LTBP1 and fibronectin 1 were found in distinct fibrillar networks. Overexpression of LTBP1 deletion constructs in osteoblast-like cells showed that N-terminal amino acids 67,467 were sufficient for incorporation into fibrillin-containing microfibrils and suggested that LTBP1 can be produced by cells distant from the site of fibril formation. In embryonic long bones in vivo, LTBP1 and fibrillin 1 colocalized at the surface of newly forming osteoid and bone. However, LTBP1-positive fibrils, which did not contain fibrillin 1, were present in cartilage matrix. These studies show that in addition to regulating TGF,1, LTBP1 may function as a structural component of connective tissue microfibrils. LTBP1 may therefore be a candidate gene for Marfan-related connective tissue disorders in which linkage to fibrillins has been excluded. [source] Spectrophotometric Analysis of Tooth Color Reproduction on Anterior All-Ceramic Crowns: Part 2: Color Reproduction and Its Transfer from In Vitro to In VivoJOURNAL OF ESTHETIC AND RESTORATIVE DENTISTRY, Issue 1 2010AKI YOSHIDA RDT ABSTRACT Color reproduction of an anterior tooth requires advanced laboratory techniques, talent, and artistic skills. Color matching in a laboratory requires the successful transfer from in vivo with careful considerations. The purpose of this study was to monitor and verify the color reproduction process for an anterior all-ceramic crown in a laboratory through spectrophotometric measurements. Furthermore, a crown insertion process using composite luting cements was assessed, and the final color match was measured and confirmed. An all-ceramic crown with a zirconia ceramic coping for the maxillary right central incisor was fabricated. There was a significant color difference between the prepared tooth and the die material. The die material selected was the closest match available. The ceramic coping filled with die material indicated a large color difference from the target tooth in both lightness and chromaticity. During the first bake, three different approaches were intentionally used corresponding with three different tooth regions (cervical, body, and incisal). The first bake created the fundamental color of the crown that allowed some color shifts in the enamel layer, which was added later. The color of the completed crown demonstrated an excellent color match, with ,E 1.27 in the incisal and 1.71 in the body. In the cervical area, color match with ,E 2.37 was fabricated with the expectation of a color effect from the underlying prepared tooth. The optimal use of composite luting cement adjusted the effect from the underlying prepared tooth color, and the color match fabricated at a laboratory was successfully transferred to the clinical setting. The precise color measurement system leads to an accurate verification of color reproduction and its transfer. CLINICAL SIGNIFICANCE The use of a dedicated dental spectrophotometer during the fabrication of an all-ceramic crown allows the dentist and the laboratory technician to accurately communicate important information to one another about the shade of the tooth preparation, the shade of the contralateral target tooth, and the influence of luting cement on the final restoration, thereby allowing the technician better control over the outcome of their tooth color matching efforts and the final color match of an all-ceramic restoration. (J Esthet Restor Dent 22:53,65, 2010) [source] 5-HT1B Receptor-Mediated Regulation of Serotonin Clearance in Rat Hippocampus In VivoJOURNAL OF NEUROCHEMISTRY, Issue 5 2000Lynette C. Daws Abstract: The 5-hydroxytryptamine (5-HT; serotonin) transporter (5-HTT) is important in terminating serotonergic neurotransmission and is a primary target for many psychotherapeutic drugs. Study of the regulation of 5-HTT activity is therefore important in understanding the control of serotonergic neurotransmission. Using high-speed chronoamperometry, we have demonstrated that local application of 5-HT1B antagonists into the CA3 region of the hippocampus prolongs the clearance of 5-HT from extracellular fluid (ECF). In the present study, we demonstrate that the 5-HT1B antagonist cyanopindolol does not produce this effect by increasing release of endogenous 5-HT or by directly binding to the 5-HTT. Dose-response studies showed that the potency of cyanopindolol to inhibit clearance of 5-HT was equivalent to that of the selective 5-HT reuptake inhibitor fluvoxamine. Local application of the 5-HT1A antagonist WAY 100635 did not alter 5-HT clearance, suggesting that the effect of cyanopindolol to prolong clearance is not via a mechanism involving 5-HT1A receptors. Finally, the effect of low doses of cyanopindolol and fluvoxamine to inhibit clearance of 5-HT from ECF was additive. These data are consistent with the hypothesis that activation of terminal 5-HT1B autoreceptors increases 5-HTT activity. [source] Stoichiometry of Tyrosine Hydroxylase Phosphorylation in the Nigrostriatal and Mesolimbic Systems In VivoJOURNAL OF NEUROCHEMISTRY, Issue 1 2000Effects of Acute Haloperidol, Related Compounds Abstract ; Electrical stimulation of the medial forebrain bundle increases 32P incorporation into striatal tyrosine hydroxylase (TH) at Ser 19, Ser31, and Ser40. In the present studies, the effects of acute haloperidol and related drugs on sitespecific TH phosphorylation stoichiometry (PS) in the nigrostriatal and mesolimbic systems were determined by quantitative blot immunolabeling using phosphorylation statespecific antibodies. The striatum (Str), substantia nigra (SN), nucleus accumbens (NAc), and ventral tegmental area (VTA) from Sprague-Dawley rats were harvested 30-40 min after a single injection of either vehicle, haloperidol (2 mg/kg), raclopride (2 mg/kg), clozapine (30 mg/kg), or SCH23390 (0.5 mg/kg). In vehicle-injected control rats, Ser19 PS was 1.5- to 2.5-fold lower in Str and NAc than in SN and VTA, Ser31 PS was two-to fourfold higher in Str and NAc than in SN and VTA, and Ser40 PS was similar between the terminal field and cell body regions. After haloperidol, Ser40 PS increased twofold in Str and NAc, whereas a smaller increase in SN and VTA was observed. The effects of haloperidol on Ser19 PS were similar to those on Ser40 in each region ; however, haloperidol treatment increased Ser31 PS at least 1.6-fold in all regions. The effects of raclopride on TH PS were comparable to those of haloperidol, whereas clozapine treatment increased TH PS at all sites in all regions. By contrast, the effects of SCH23390 on TH PS were relatively small and restricted to the NAc. The stoichiometries of site-specific TH phosphorylation in vivo are presented for the first time. The nigrostriatal and mesolimbic systems have common features of TH PS, distinguished by differences in TH PS between the terminal field and cell body regions and by dissimilar increases in TH PS in the terminal field and cell body regions after acute haloperidol. [source] Local Corticosterone Infusion Enhances Nocturnal Pineal Melatonin Production In VivoJOURNAL OF NEUROENDOCRINOLOGY, Issue 2 2009P. A. C. M. Fernandes Melatonin, an important marker of the endogenous rhythmicity in mammals, also plays a role in the body defence against pathogens and injuries. In vitro experiments have shown that either pro- or anti-inflammatory agents, acting directly in the organ, are able to change noradrenaline-induced pineal indoleamine production. Whereas corticosterone potentiates melatonin production, incubation of the gland with tumour necrosis factor-, decreases pineal hormonal production. In the present study, we show that nocturnal melatonin production measured by intra-pineal microdialysis is enhanced in pineals perfused with corticosterone at concentrations similar to those measured in inflamed animals. In vitro experiments suggest that this enhancement may be due to an increase in the activity of the two enzymes that convert serotonin to N -acetylserotonin (NAS) and NAS to melatonin. The present results support the hypothesis that the pineal gland is a sensor of inflammation mediators and that it plays a central role in the control of the inflammatory response. [source] KiSS-1 and GPR54 Genes are Co-Expressed in Rat Gonadotrophs and Differentially Regulated In Vivo by Oestradiol and Gonadotrophin-Releasing HormoneJOURNAL OF NEUROENDOCRINOLOGY, Issue 3 2008N. Richard Kisspeptin, the product derived from KiSS-1, and its cognate receptor, GPR54, both exert a role in the neuroendocrine control of reproduction by regulating gonadotrophin-releasing hormone (GnRH) secretion. In the present study, we demonstrate, using dual immunofluorescence with specific antibodies, that the KiSS-1 and GPR54 genes are both expressed in rat gonadotrophs. All luteinising hormone ,-immunoreactive (LH,-ir) cells were stained by the KiSS-1 antibody but some kisspeptin-ir cells were not LH, positive; thus, we cannot exclude the possibility that kisspeptins are expressed in other pituitary cells. All GPR54-ir are co-localised with LH, cells, but only a subset of LH, cells are stained with the GPR54 antibody. Using the real-time reverse transcription-polymerase chain reaction (RT-PCR), we found that the expression of KiSS-1 and GPR54 is differentially regulated by steroids. In the female, KiSS-1 mRNA levels dramatically decreased following ovariectomy (OVX), and this decrease was prevented by administration of 17,-oestradiol (E2), but not by administration of GnRH antagonist or agonist. Administration of E2 in OVX rats receiving either GnRH antagonist or agonist clearly shows that E2 acts directly on the pituitary to positively control KiSS-1 expression. In OVX rats, administration of the selective oestrogen receptor (ER), ligand propylpyrazoletriol, but not the selective ER, ligand diarylpropionitrile, mimics this effect. By contrast, our study shows that GPR54 expression is positively regulated by GnRH and negatively controlled by chronic exposure to E2. In summary, our data document for the first time that, in the female rat pituitary, KiSS-1 expression is up-regulated by oestradiol, similarly to that seen in the anteroventral periventricular nucleus of the hypothalamus. Conversely, GPR54 is up-regulated by GnRH, which exclusively targets gonadotrophs. [source] Sex Differences in Oestrogen-Induced p44/42 MAPK Phosphorylation in the Mouse Brain In VivoJOURNAL OF NEUROENDOCRINOLOGY, Issue 8 2006K. Barabás In addition to the classical direct genomic mechanisms of action, oestrogen also exerts poorly understood, nonclassical effects on the signalling system in neurones. In the present study, we investigated whether sex differences exist in gonadectomy- and oestrogen-induced effects on p44/42 mitogen-activated protein kinase (MAPK) phosphorylation in specific brain regions of mice. We demonstrate that MAPK immunoreactivity was not altered by gonadectomy or oestrogen treatment in either sex. However, we show that the level of phosphorylated MAPK (pMAPK) within the anteroventral periventricular nucleus (AVPV) was consistently higher in males than females irrespective of gonadal steroid hormone status. In addition, gonadectomy was found to decrease pMAPK immunoreactivity within the piriform cortex of males. Oestrogen increased pMAPK immunoreactivity in the medial preoptic area and AVPV of females, but failed to have the same effect in male mice. Overall, these results demonstrate a marked sex difference in oestrogen-induced alteration of MAPK phosphorylation in the brain in vivo. [source] Mechanical Behavior and Failure Analysis of Prosthetic Retaining Screws after Long-term Use In Vivo.JOURNAL OF PROSTHODONTICS, Issue 3 2008Part 1: Characterization of Adhesive Wear, Structure of Retaining Screws Abstract Purpose: The general aim of this study and those presented in Parts 2,4 of this series was to characterize the structure, properties, wear, and fracture of prosthetic retaining screws in fixed detachable hybrid prostheses after long-term use in vivo. This part of the overall investigation addresses whether there are differences in thread wear between the screws closest to the fulcrum and those that are farthest from the fulcrum in fixed detachable hybrid prostheses. Materials and Methods: The total number of prosthetic retaining screws used in this study was 100 (10 new and 90 used). New screws (controls) from Nobel Biocare (NB) were divided into Group 1 (slotted) and Group 2 (hexed). Ninety used screws (in service 18,120 months) were retrieved from fixed detachable hybrid prostheses in 18 patients (5 screws from each patient, 60 from NB and 30 from Sterngold). The used screws were divided into 18 groups. Additionally, each group was subdivided into A and B categories. Category A contained the middle three prosthetic screws, which were considered the farthest screws from the fulcrum line. Category B contained the most posterior two screws, which were considered the screws closest to the fulcrum line. All 100 screws were subjected to thorough, nondestructive testing. Results: Light and scanning electron microscopic examination of all used screws for each group revealed surface deterioration of the active profile of the screw threads consistent with adhesive wear. The observed thread profile deterioration ranged from mild to severe. The wear was aggressive enough to cause galling, which led to thinning of the threads and, in severe cases, to knife-edges at thread crests. In ten groups, the most anterior three screws exhibited more wear than the most posterior two screws. In addition to thread wear, severe plastic deformation was detected on the bottom part of each screw for three groups, and a long external longitudinal crack was detected in one screw of Group 2. Conclusions: The findings of this study and those presented in Parts 2,4 demonstrate that different retaining screws from the same manufacturer and/or from different manufacturers have different geometrical design, microstructures, major alloy constituents, and microhardness, and that these differences influence their preload and fractured load values. In this part of the overall investigation, the occurrence of galling as a result of wear involving prosthetic retaining screws appears to be an inevitable and unavoidable consequence of long-term use in vivo in fixed detachable hybrid prostheses regardless of the intended/original preload value. The galling rate is greater on the middle three screws compared to the most posterior two screws in fixed detachable hybrid prostheses. The wear pattern is consistent with an adhesive wear mechanism; however, this study does not provide enough data to support a definitive analysis. [source] Mechanical Behavior and Failure Analysis of Prosthetic Retaining Screws after Long-Term Use In Vivo.JOURNAL OF PROSTHODONTICS, Issue 3 2008Microhardness Analysis, Part 2: Metallurgical Abstract Purpose: This study involved testing and analyzing multiple retrieved prosthetic retaining screws after long-term use in vivo to: (1) detect manufacturing defects that could affect in-service behavior; (2) characterize the microstructure and alloy composition; and (3) further characterize the wear mechanism of the screw threads. Materials and Methods: Two new (control) screws from Nobel Biocare (NB) and 18 used (in service 18,120 months) retaining screws [12 from NB and 6 from Sterngold (SG)] were: (1) metallographically examined by light microscopy and scanning electron microscopy (SEM) to determine the microstructure; (2) analyzed by energy dispersive X-ray (EDX) microanalysis to determine the qualitative and semiquantitative average alloy and individual phase compositions; and (3) tested for Vickers microhardness. Results: Examination of polished longitudinal sections of the screws using light microscopy revealed a significant defect in only one Group 4 screw. No significant defects in any other screws were observed. The defect was considered a "seam" originating as a "hot tear" during original casting solidification of the alloy. Additionally, the examination of longitudinal sections of the screws revealed a uniform homogeneous microstructure in some groups, while in other groups the sections exhibited rows of second phase particles. The screws for some groups demonstrated severe deformation of the lower threads and the bottom part of the screw leading to the formation of crevices and grooves. Some NB screws were comprised of Au-based alloy with Pt, Cu, and Ag as alloy elements, while others (Groups 4 and 19) were Pd-based with Ga, Cu, and Au alloy elements. The microstructure was homogeneous with fine or equiaxed grains for all groups except Group 4, which appeared inhomogeneous with anomalous grains. SG screws demonstrated a typical dendritic structure and were Au-based alloy with Cu and Ag alloy elements. There were differences in the microhardness of gold alloy screws from NB and SG as well as palladium alloy screws from NB. Conclusions: Significant differences within NB retaining screws and between NB and SG screws were found for microstructure, major alloy constituents, and microhardness. [source] Dental Materials In Vivo: Aging and Related PhenomenaJOURNAL OF PROSTHODONTICS, Issue 2 2004Mary P. Walker DDS No abstract is available for this article. [source] Peroxisome Proliferator-Activated Receptors (PPAR) and the Mitochondrial Aldehyde Dehydrogenase (ALDH2) Promoter In Vitro and In VivoALCOHOLISM, Issue 7 2001David W. Crabb Background : The aldehyde dehydrogenase 2 (ALDH2) promoter contains a nuclear receptor response element (NRRE) that represents an overlapping direct repeat-1 (DR-1) and -5 (DR-5) element. Because DR-1 elements are preferred binding sites for peroxisome proliferator-activated receptors (PPARs), we tested the hypothesis that PPARs regulate ALDH2 expression. Methods: We examined the ability of PPAR isoforms to bind to the ALDH2 NRRE in electrophoretic mobility shift assays, their ability to activate the transcription of promoter-reporter constructs containing this NRRE, the effect of PPAR ligands on ALDH2 expression in liver, and the role of the PPAR, on the expression of ALDH2 by using PPAR,-null mice. Results: In vitro translated PPARs bound the ALDH NRRE with high affinity. Mutation of the NRRE indicated that binding was mediated by the DR-1 element. Cotransfection of PPAR expression plasmids showed that PPAR, had no effect on expression of heterologous promoter constructs containing the NRRE. PPAR, slightly induced expression, whereas PPAR, repressed basal activity of the promoter and blocked induction by hepatocyte nuclear factor 4. Treatment of rats with the PPAR ligand clofibrate repressed expression of ALDH2 in rats fed either stock rodent chow or a low-protein diet. Consistent with the transfection data, expression of ALDH2 protein was not different in PPAR,-null mice. Treatment of the mice with the PPAR, agonist WY14643 slightly decreased the level of ALDH2 protein in both wild-type and PPAR,-null mice, suggesting that the effect of WY14643 was not mediated by the receptor. Conclusions: These data indicate that ALDH2 is not part of the battery of lipid metabolizing enzymes and proteins regulated by PPAR, [source] Hyperglycemia Stimulates a Sustained Increase in Hydraulic Conductivity In Vivo without Any Change in Reflection CoefficientMICROCIRCULATION, Issue 7 2007RACHEL M. PERRIN ABSTRACT Objective: Increased microvascular permeability contributes to the development of diabetic microvascular complications and diabetic vasculopathy is correlated with blood glucose levels. The mechanisms underlying increased permeability, however, are poorly understood. Methods: The Landis-Michel technique was used to measure water permeability (hydraulic conductivity, Lp) and macromolecular permeability (reflection coefficient, ,) of exchange capillaries in frogs and rats. Results: Dialysed normoglycemic plasma from diabetic patients had no effect on Lp. The same plasma with 20 mM glucose increased hydraulic conductivity from (mean ± SEM × 10,7 cm · s,1· cm H2O,1) 5.73 ± 2.01 to 13.09 ± 2.67 (P < .01). Nondiabetic control plasma did not affect Lp, but addition of 20 mM glucose increased Lp to a similar degree. The effect of glucose alone was examined. Glucose at 20 mM increased Lp, from 2.82 ± 0.61 to 4.71 ± 1.35 × 10, 7 cm · s, 1· cm H2O,1 (P = .002, n = 13). A similar increase was seen in rat mesenteric microvessels, from 1.04 ± 0.40 in control perfusions to 2.18 ± 0.56, P < .05. The microvascular macromolecular reflection coefficient in all the above experiments was unaltered. The use of specific inhibitors indicated that the glucose-induced increased Lp did not appear to be mediated through protein kinase C (PKC), free radical generation, glucose metabolism, or albumin glycation. Conclusions: These data suggest that hyperglycemia induced increased apparent protein permeability may be secondary to a glucose-mediated change in macromolecular convective flux rather than any change in protein permeability per se. The authors speculate that the increased microvascular permeability to water in vivo is mediated by direct interaction of glucose with the endothelial cells (perhaps with the glycocalyx). [source] Differential Roles of CD36, ICAM-1, and P-selectin in Plasmodium falciparum Cytoadherence In VivoMICROCIRCULATION, Issue 6 2007Bryan G. Yipp ABSTRACT Cytoadherence of Plasmodium falciparum -infected red blood cells (IRBCs) on human microvascular endothelium is mediated by synergistic adhesive interactions with different adhesion molecules in vitro. Here, the authors used a unique human/severe combined immunodeficient (SCID) mouse chimeric model to directly visualize IRBC,endothelial interactions in an intact human microvasculature in vivo. Stimulation of human skin grafts with 100 ng TNF-, for 4 h led to a dramatic reduction in the distance rolled by IRBCs before arrest, so that the majority of IRBCs adhered directly to the endothelium with a 1.8-fold increase in the number of adherent cells. The decrease in rolling distance and increase in adhesion could be reversed by anti-ICAM-1. More importantly, the effect of TNF-, could be seen only in the presence of CD36. A further increase in adhesion by 4.9-fold was observed after 24 h of TNF-, stimulation. The increase could be reversed by anti-ICAM-1, but not anti-VCAM-1. In histamine-stimulated grafts, the rolling flux fraction and adhesion increased by 2.8- and 1.6-fold, respectively. The increases were attributable to P-selectin as an inhibitory anti-P-selectin antibody abrogated both the increased rolling flux fraction and firm adhesion. These findings indicate that in addition to CD36, ICAM-1, and P-selectin are major contributors to the dynamic process of IRBC adhesion by different mechanisms in vivo. [source] Microvascular Thrombosis Models in Venules and Arterioles In VivoMICROCIRCULATION, Issue 3 2005ROLANDO E. RUMBAUT MD ABSTRACT Platelets are intimately involved in hemostasis and thrombosis. Under physiological conditions, circulating platelets do not interact with microvascular walls. However, in response to microvascular injury, platelet adhesion and subsequent thrombus formation may be observed in venules and arterioles in vivo. Numerous intravital video microscopy techniques have been described to induce and monitor the formation of microvascular thrombi. The mechanisms of microvascular injury vary widely among different models. Some models induce platelet activation with minimal effects on endothelium, others induce endothelial inflammation or injury, while other models lead to thrombus formation associated with endothelial denudation. The molecular mechanisms mediating platelet,vessel wall adhesive interactions differ among various models. In some instances, differences in responses between venules and arterioles are described that cannot be explained solely by hemodynamic factors. Several models for induction of microvascular thrombosis in vivo are outlined in this review, with a focus on the mechanisms of injury and thrombus formation, as well as on differences in responses between venules and arterioles. Recognizing these characteristics should help investigators select an appropriate model for studying microvascular thrombosis in vivo. [source] Absorption Spectra of Human Skin In Vivo in the Ultraviolet Wavelength Range Measured by OptoacousticsPHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 1 2009Merve Meinhardt Knowledge of the optical properties of human skin in the ultraviolet range is fundamental for photobiologic research. However, optical properties of human skin in the ultraviolet spectral range have so far mainly been measured ex vivo. We have determined the absorption spectra of human skin in vivo in the wavelength range from 290 to 341 nm in 3 nm steps using laser optoacoustics. In this technique, optical properties are derived from the pressure profile generated by absorbed light energy in the sample. In a study on 20 subjects belonging to phototypes I,IV, we studied the optical properties at the volar and dorsal aspect of the forearm as well as on the thenar. Analysis of the measured absorption spectra shows that comparable skin areas,like different sides of the forearm,have qualitatively similar optical characteristics. Still, the optical properties may vary substantially within the same area, probably due to the skin structure and inhomogeneities. Comparison of the spectra from different skin sites indicates that the spectral characteristics of the stratum corneum and its chromophores play an important role for the optical properties of human skin in vivo in the ultraviolet B range. [source] Optical Properties of Human Melanocytic Nevi In VivoPHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 1 2009George Zonios We present an in vivo study of the optical properties of melanin present in melanocytic nevi of human subjects with Fitzpatrick skin type III (Caucasian descent) using optical spectroscopy. We show that the melanin absorption spectrum exhibits an exponential dependence on wavelength with a decay constant which follows a normal distribution characteristic of a random biological variable. Moreover, we demonstrate lack of correlation among melanin optical properties, melanin concentration and skin light scattering properties, which indicates that the true optical absorption of melanin can be measured free from confounding scattering effects. We also show that the average melanin absorption spectrum in vivo is in very good agreement with a previously reported oxygen photoconsumption action spectrum of melanin. Finally, we provide an overview of the emerging picture of the melanin absorption properties in vivo among various skin types and also among various skin lesions such as melanocytic nevi and melanoma. [source] Polychromatic Light Similar to the Terrestrial Solar Spectrum Without its UV Component Stimulates DNA Synthesis in Human Peripheral Blood Lymphocytes In Vivo and In VitroPHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 5 2006Natalya A. Zhevago ABSTRACT Immunosuppressive effects of the minor component of the terrestrial solar spectrum, UV radiation, have been substantiated over the past several years. This raises the question of what influence the dominant part of the solar spectrum,visible and IR light,would have on the human immune system. In the present randomized, placebo-controlled double-blind study a small area of the body surface of volunteers was irradiated with polychromatic light (480,3400 nm), simulating the significant part of the terrestial sunlight irradiance spectrum and its power density. An average 2.5-fold to three-fold increase in spontaneous and phytohemagglutinin-induced DNA synthesis in peripheral blood lymphocytes (Lym) was revealed at 0.5,24 h after irradiation at a therapeutic dose (12 J/cm2) in subjects with low preirradiation levels of both processes. The in vivo findings were echoed in parallel in vitro experiments, when blood drawn from the same subjects was directly irradiated (2.4 J/cm2), or when the irradiated blood was mixed 1:10 with nonirradiated autolo-gous blood to model events in the circulation following transcutaneous blood photomodification. Our data suggest that exposure of the human body to polychromatic visible + IR light may photomodify blood in the dermal vasculature of the irradiated area to lead to an immediate transfer of the light-induced effects to Lym of the entire circulating blood, which can result in modulation of Lym functional state at the systemic level. [source] Imaging of Photodynamically Generated Singlet Oxygen Luminescence In Vivo,PHOTOCHEMISTRY & PHOTOBIOLOGY, Issue 4 2005Mark J. Niedre ABSTRACT We describe a novel scanning-laser system for imaging type-II photodynamically generated singlet oxygen (1O2[1,g]) luminescence and demonstrate it in vivo in an intradermal tumor model in mice. We verify the strong oxygen-dependence of the signal and show that the images are near the practical resolution limit. [source] | |||||||||||||||||||