Effect of calcium ions on rat osseous plate alkaline phosphatase activity

Rat osseous plate alkaline phosphatase is a metalloenzyme with two binding sites for Zn2+ (sites I and III) and one for Mg2+ (site II). This enzyme is stimulated synergistically by Zn2+ and Mg2+ (Ciancaglini et al., 1992) and also by Mn2+ (Leone et al., 1995) and Co2+ (Ciancaglini et al., 1995). This study was aimed to investigate the modulation of enzyme activity by Ca2+. In the absence of Zn2+ and Mg2+, Ca2+ had no effects on the activity of Chelex-treated, Polidocanol-solubilized enzyme. However, in the presence of 10 mu M MgCl2, increasing concentration of Ca2+ were inhibitory, suggesting the displacement of Mg2+ from the magnesium-reconstituted enzyme. For calcium-reconstituted enzyme, Zn2+ concentrations Zip to 0.1 mu M were stimulatory, increasing specific activity from 130 U/mg to about 240 U/mg with a K-0.5 = 8.5 nM. Above 0.1 mu M Zn2+ exerted a strong inhibitory effect and concentrations of Ca2+ up to I mM were not enough to counteract this inhibition, indicating that Ca2+ was easily displaced by Zn2+. At fixed concentrations of Ca2+, increasing concentrations of Mg2+ increased the enzyme specific activity from 472 U/mg to about 547 U/mg, but K-0.5 values were significantly affected (from 4.4 mu M to 38.0 mu M). The synergistic effects observed for the activity of Ca2+ plus magnesium-reconstituted enzyme, suggested that these two ions bind to the different sites. A model to explain the effect of Ca2+ on the activity of the enzyme is presented. (C) 1997 Elsevier B.V.

OSSEOUS PLATE ALKALINE-PHOSPHATASE IS ANCHORED BY GPI

Alkaline phosphatase activity was released up to 100% from the membrane by using 0.1 U of phosphatidylinositol-specific phospholipase C from B. thuringiensis. The Mr of solubilized enzyme was 145,000 by Sephacryl S-300 gel filtration and 66,000 by SDS-PAGE, suggesting a dimeric structure. Solubilization of the membrane-bound enzyme with phospholipase C did not destroy its ability to hydrolyze p-nitrophenyl phosphate (PNPP) (264.3 mu mol min(-1) mg(-1)), ATP (42.0 mu mol min(-1) mg(-1)) and pyrophosphate (28.4 mu mol min(-1) mg(-1)). The hydrolysis of ATP and PNPP by solubilized enzyme exhibited ''Michaelian'' kinetics with K-0.5 = 70 and 979 mu M, respectively. For pyrophosphate, K-0.5 was 128 mu M and site-site interactions were observed (n = 1.4). Magnesium ions were stimulatory (K-d = 1.5 mM) but zinc ions were powerful non-competitive inhibitors (K-d = 6.2 mu M) of solubilized enzyme. Treatment of solubilized alkaline phosphatase with Chellex 100 reduced the original PNPPase activity to 5%. Cobalt (K-0.5 = 10.1 mu M), magnesium (K-0.5 = 29.5 mu M) and manganese ions (K-0.5 = 5 mu M) restored the activity of the apoenzyme with positive cooperativity, suggesting that phosphatidylinositol-specific phospholipase C-solubilized alkaline phosphatase is a metalloenzyme. The stimulation of the apoenzyme by calcium ions (K-0.5 = 653 mu M) was lower than that observed for the other ions (26%) and exhibited site-site interactions (n = 0.7). Zinc ions had no effect on the apoenzyme of the solubilized enzyme.

KINETIC-PROPERTIES OF OSSEOUS PLATE ALKALINE-PHOSPHATASE FROM DIABETIC RATS

1. Increased levels of bone alkaline phosphatase activity were observed in diabetic rats. These animals exhibited impaired bone development without concomitant alterations of the sequence of cellular transformations.2. Alkaline phosphatase activity was delayed in diabetic rats but the kinetic parameters for the hydrolysis of p-Nitrophenylphosphate (PNPP) were virtually the same observed for controls (N = 1.2 and K0.5 = 43 muM).3. Alkaline phosphatase from diabetic rats had a better affinity (K0.5 = 38 muM) for magnesium ions than controls (K0.5 = 9 1 muM).4. Zinc ions affected alkaline phosphatase activity from control and diabetic rats in the same way (K0.5 = 10 muM).

Osseous regeneration in the presence of fibrin adhesive material (Tissucol) and epsilon-aminocaproic acid (EACA).

The effects of Tissucol and Tissucol/EACA on bone healing were evaluated histologically. Experimental defects were made in both tibias of 25 rats. Test materials were placed in defects in right tibias and left tibias served as control. Five animals in each group were killed at 1, 3, 7, 14 and 21 days after surgery. Results showed that: a) Tissucol did not interfere with connective and osseous tissue formation; b) Tissucol allowed new bone formation; c) Tissue residues in Tissucol groups in sections of 21-day specimens did not impair healing; d) Tissucol/EACA was usually completely resorbed and healing was complete 21 days after surgery in the Tissucol/EACA group.

A unique intraorbital osseous structure in the large fruit-eating bat (Artibeus lituratus)

Objective: To describe the normal bony orbital structure of the large fruit-eating bat (Artibeus lituratus) with emphasis on a unique intraorbital bony structure previously not described in the literature. Procedures: The bony anatomy of the orbital cavity was studied on dissected skulls of large fruit-eating bats. The anatomic description of a unique intraorbital spine was made while studying the bony orbit of macerated skulls. Additional observations were made on dissected formalin-fixed whole heads. Both procedures were performed under a stereo dissecting microscope, using ×2-4-magnification. A histologic analysis of soft tissues surrounding this cylindrical bony structure was performed using cross and longitudinal oblique sections from decalcified whole heads, which had been fixed in formalin. Additionally, biometric measurements and a histomorphometric analysis were performed. Results and conclusions: An intraorbital cylindrical osseous structure measuring 3.96 ± 0.68 mm in length and 155.62 ± 14.03 μm in diameter was observed in the large fruit-eating bat (A. lituratus), creating a unique orbital structural design among mammals. We suggest the name optic spine of the alisphenoid bone. The anatomic, biometric and histologic characterization of this element might contribute to a further understanding of the dynamics of bat vision and the sort of factors that influenced evolution of the visual system of microbats. The authors hope that the documentation of this distinctive anatomic feature will also expand the debate about the phylogenetic analysis of the relationship among bat species in the near future. © 2007 American College of Veterinary Ophthalmologists.

Effects of fibrin sealer and resorbable gelatin on the repair of osseous defects in rat tibia

Gelfoam® - a biologically resorbable gelatin sponge - has the function of restricting hemorrhage, providing platelet rupture, and supporting fibrin threads. Beriplast® - a fibrinogen-thrombin compound - is used to adhere tissues, to consolidate sutures and in hemostasis. The objective of this study was to perform a histological analysis of the effects of haemostatic agents on osseous repair. These materials were inserted into surgical sites in young rat right and left tibiae. After the observation periods of 7, 14, 30 and 45 days, according to the bioethic protocol, the animals were killed, the tibiae were removed and fixed in 10% formalin and decalcified in equal parts of formic acid and sodium citrate solutions. After routine processing, the specimens were embedded in paraffin for microtomy. Analysis of the results demonstrated that the haemostatic agents are effective in controlling hemorrhage; they stimulate osteogenesis, featuring a pattern of osseous tissue formation similar to the control pattern, although the amount of osseous trabeculae was superior, especially in the Gelfoam group in the periods of 7 and 14 days; 30 days after surgery, the delay in tissue healing in the control group in relation to the experimental groups started to decrease, and the control and experimental groups exhibited similar tissue repair after 45 days, when all the groups exhibited secondary osseous tissue.

Florid cemento-osseous dysplasia: A report of three cases

Florid cemento-osseous dysplasia (FCOD) is a non-neoplastic condition of the jaws that Is not associated with inflammation of the pulp or periodontal tissue. This article reports on three cases that were diagnosed as FCOD, demonstrating the importance of both clinical and radiographic diagnosis and the clinical management of these lesions.

Avaliação do padrão de metilação dos genes WT1 e RARß em metaplasia intestinal e associação com infecção pela Helicobacter pylori

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Expressão de genes relacionados ao ciclo celular e proteção da mucosa gástrica em metaplasia intestinal e ulcera gástrica em comparação com câncer gástrico

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Deregulation of Annexin-A1 and Galectin-1 Expression in Precancerous Gastric Lesions: Intestinal Metaplasia and Gastric Ulcer

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Characterization of the phosphatidylinositol-specific phospholipase C-released form of rat osseous plate alkaline phosphatase and its possible significance on endochondral ossification

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Florid cemento-osseous dysplasia: a report of three cases

Florid cemento-osseous dysplasia (FCOD) is a non-neoplastic condition of the jaws that is not associated with inflammation of the pulp or periodontal tissue. This article reports on three cases that were diagnosed as FCOD, demonstrating the importance of both clinical and radiographic diagnosis and the clinical management of these lesions.

Aggressive osseous commitment result by keratocyst odontogenic tumour: case report, radiographic and clinical standpoints

Keratocystic odontogenic tumour (KCOT) previously known as odontogenic keratocyst was recently classified as a benign lesion characterized by an infiltrating pattern, local aggressiveness with the propensity to recurrence. It is thought to arise from the dental lamina. Pain is usually not associated with KCOT until swelling occurs, and it commonly affects the posterior mandible. Multiple KCOT are associated with nevoid basal cell carcinoma syndrome. This study reports an aggressive case of KCOT with destruction of the osseous tissue of the mandible, accentuated face asymmetry, dysphagia and dysphonia. It was managed with a defined protocol which entailed diagnosis, treatment with enucleation along with peripheral ostectomy and rehabilitation. A long-term follow-up schedule was provided to the patient to observe the recurrence behaviour of this cyst. In postoperative phase, no complication was noticed regarding wound healing and recurrence.

Interação entre Citocinas, Plaquetas e Fibrogênese na Investigação da Metaplasia Mielóide Hepática

Pós-graduação em Pesquisa e Desenvolvimento (Biotecnologia Médica) - FMB